March 1978
HEALTH IMPLICATIONS OF
SEWAGE TREATMENT FACILITIES
by
D. E. Johnson, D. E. Camann, J. W. Register,
R. J. Prevost, J. B. Tillery, R. E. Thomas,
J. M. Taylor, and J. M. Hosenfeld
Southwest Research Institute
San Antonio, Texas 78284
Contract No. 68-02-1746
Project Officer
Herbert R. Pahren
Field Studies Division
Health Effects Research Laboratory
Cincinnati, Ohio 45268
HEALTH EFFECTS RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
I. S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI. OHIO 45268
SOUTHWEST RESEARCH INSTITUTE
SAN ANTONIO CORPUS CHRISTI HOUSTON
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March 1978
HEALTH IMPLICATIONS OF
SEWAGE TREATMENT FACILITIES
by
D. E. Johnson, D. E. Camann, J. W. Register,
R. J. Prevost, J. B. Tillery, R. E. Thomas,
J. M. Taylor, and J. M. Hosenfeld
Southwest Research Institute
San Antonio, Texas 78284
Contract No. 68-02-1746
Project Officer
Herbert R. Pahren
Field Studies Division
Health Effects Research Laboratory
Cincinnati, Ohio 45268
HEALTH EFFECTS RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U. S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
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DISCLAIMER
This report has been reviewed by the Health Effects Research Laboratory, U.S. Environmental Protec-
tion Agency, and approved for publication. Approval does not signify that the contents necessarily reflect the
views and policies of the U.S. Environmental Protection Agency, nor does mention of trade names or com-
mercial products constitute endorsement or recommendation for use.
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FOREWORD
The U.S. Environmental Protection Agency was created because of increasing public and government
concern about the dangers of pollution to the health and welfare of the American people. Noxious air, foul
water, and spoiled land are tragic testimony to the deterioration of our natural environment. The complexity
of that environment and the interplay between its components require a concentrated and integrated attack on
the problem.
Research and development is that necessary first step in problem solution and it involves defining the
problem, measuring its impact, and searching for solutions. The primary mission of the Health Effects
Research Laboratory in Cincinnati (HERL) is to provide a sound health effects data base in support of the
regulatory activities of the EPA. To this end, HERL conducts a research program to identify, characterize,
and quantitate harmful effects of pollutants that may result from exposure to chemical, physical, or
biological agents found in the environment. In addition to valuable health information generated by these ac-
tivities, new research techniques and methods are being developed that contribute to a better understanding of
human biochemical and physiological functions, and how these functions are altered by low-level insults.
This report provides an assessment and discussion of the health of persons living near a wastewater treat-
ment plant. With a better understanding of the health effects, measures can be developed to reduce exposure
to potentially harmful materials.
R. J. Garner
Director
Health Effects Research Laboratory
in
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ABSTRACT
It is known that aerosols containing microorganisms and trace metals are emitted from wastewater treat-
ment facilities. Virtually nothing was known about the possible health effects on populations living near these
operations. In this study, environmental monitoring, household health survey, and sampling for clinical
specimens of human subjects were conducted within a 5 kilometer distance from a wastewater treatment plant
near Chicago, Illinois. The residential area began 400 meters from the plant.
Although the treatment plant was a source of indicator bacteria, coliphage, pathogenic bacteria,
enteroviruses, and mercury in the aerosols emanating from its aeration basins, the levels of microbiological
and chemical agents of the air, water, and soil samples in the neighboring residential areas were not
distinguishable from the background levels.
From the patterns observed in the household health survey, the increased incidence of skin disease, and
the symptoms of nausea, vomiting, general weakness, diarrhea, and pain in chest on deep breathing may be
associated with the nearby operation of the wastewater treatment plant.
Although of little practical health significance, alpha-and gamma-hemolytic streptococcus isolations in
throat cultures of nearby residents may be related to plant operations. In contrast, 31 viral antibody tests and
attempted isolations of many pathogenic bacteria, parasites, and viruses yielded no evidence of an adverse
wastewater treatment plant effect.
Overall, the findings did not detect a health hazard for persons living beyond 400 meters from the well-
operated wastewater treatment plant.
This report was submitted in fulfillment of Contract No. 68-02-1746 by Southwest Research Institute
under the sponsorship of the U.S. Environmental Protection Agency. This report covers the period July 1974
to October 1976, and work was completed as of October 1977.
IV
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CONTENTS
Page
FIGURES .,..., vi«
TABLES ..... ix
SECTION 1. INTRODUCTION ! 1
MOTIVATION AND PURPOSE OF THE STUDY 1
DESIGN OF THE STUDY . 2
RESEARCH QUESTIONS , ... 6
INTERPRETATION OF FINDINGS , 7
PARTICIPATING ORGANIZATIONS AND PRINCIPAL PERSONNEL 7
SECTION 2. CONCLUSIONS '. 10
SECTION 3. RECOMMENDATIONS. ...... 11
SECTION 4. THE STUDY SITE. ....... .. .... 12
SITE SELECTION PROCESS 12
Site Requirements 12
Site Visit , ;13
THE SELECTED STUDY SITE . . . 13
John E. Egan Water Reclamation Plant 13
The Study Area 15
SECTION 5. METHODS 1.9
HOUSEHOLD SURVEY AND PARTICIPANT RECRUITMENT 19
General Design 19
Questionnaire Development 19
Survey Design 26
Performance of Baseline; Additional Baseline and Operational Household Surveys .. 27
ENVIRONMENTAL SAMPLING , 28
Organization Overview ., .-.-..., . . .. . 28
Air Samples 29
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CONTENTS (Cont'd)
Mixed Liquor Samples from Aeration Basin 37
Soil Samples 37
Water Samples 38
Meteorological Sampling 38
CLINICAL SPECIMEN SAMPLING 39
Organizational Overview 39
Presampling Preparation 39
Sampling Protocol and Sample Collection 40
Sample Processing 43
ANALYTICAL METHODS 45
Organizational Overview 45
Microbiological Analysis of Environmental Samples 45
Trace Metal Analysis of Environmental Samples 53
Microbiological Analysis of Clinical Specimens 59
Chemical Analysis of Human Tissue Samples 60
DATA FLOW AND PROCESSING 64
Identification and Integrity of Sample Data 64
Sample Labels 65
Data Forms and Reporting System 65
Participant and Household Survey Data Bases 69
Computational Techniques 69
GENERAL STATISTICAL METHODOLOGY 69
SECTION 6. RESULTS 74
ENVIRONMENTAL MONITORING 74
Aeration Basin Samples 74
Air Samples 79
Soil Samples 102
Water Samples 112
HOUSEHOLD HEALTH SURVEY 120
Household and Personal Characteristics 120
Analysis of Health Survey Illness, Disease and Symptom Incidence 129
HUMAN SUBJECTS CHARACTERISTICS 152
Human Subject Participation by Sampling Period 152
Distribution of Participants Sampled in October 1976 153
Distributions of 100 Human Subjects Analyzed for Additional Viral Serology 160
v;
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CONTENTS (Cont'd)
CLINICAL SPECIMEN MICROBIOLOGY.. 162
Statistical Methodology 162
Bacterial Isolations 166
Parasite Isolations I" 173
Viral Isolations 174
Viral Antibody Titer 176
CLINICAL SPECIMEN TRACE METALS 191
HUMAN SUBJECT CURRENT HEALTH STATUS ... . 1 ......: . 210
SECTION 7. DISCUSSION .-;•:... 216
APPENDICES ...............: 231
A JUSTIFICATION OF SURVEY QUESTIONNAIRE FORMS . 231
B SURVEY BACKUP MATERIALS . . 255
C LETTERS OF ACCEPTANCE/NON-ACCEPTANCE TO
SURVEY PARTICIPANTS 261
D VOLUNTEER'S INFORMED CONSENT FORM . ... 264
E INSTRUCTION SHEET 267
F CERTIFICATION OF APPRECIATION .-...- :...... 270
G TYPICAL CODED DATA REPORTING FORMS 272
H INDIVIDUAL PARTICIPANT DATA TABLES 288
vn
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FIGURES
Figure Page
1. John E. Egan Water Reclamation Plant flow diagram 14
2. Map of study area. 16
3. Household health survey questionnaire 20
4. Health survey participant questionnaire 22
5. Current health status questionnaire 24
6. Air sampler location design 30
7. Schematic diagram of mycobacteria isolation 48
8. Schematic diagram of enterics isolation 50
9. Apparatus for the acid digestion of air samples 56
10. Typical sample labels 67
11. General statistical analysis methodology 72
12. Distribution of daily wind directions during the operational period (Jan-Oct, 1976) 83
13. Cadmium concentrations plotted by distance 98
14. Lead concentrations plotted by distance 99
15. Mercury concentrations plotted by distance 100
VIH
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TABLES
Table
1. Environmental and Human Subject Sampling Matrices.....; .:;...•...;'.......;.... 5
2. Participating Organizations and Key Personnel ...........'. 8
3. Age-Sex Breakdown of Population Surrounding John Egan Plant (1970 Census) 18
4. Initial Participants Recruited in September 1974 ...:... 27
5. Additional Participants Recruited in September 1975 . 28
6. Key to Stability Categories 33
7. The Fifteen-Digit SampleCode . ..•'. ......... 66
8. Automated Computing Procedures '.•;.' ....'...;....•. 70
9. UTSA Microbial Characterization of Period 5 Aeration Basin Grab Samples 75
10. Microbiological Concentrations in Period 4 Aeration Basin Samples with High Volume
Aerosol Sampling Runs 76
11. Microbiological Concentrations in Period 5 First Stage Aeration Basin Grab Samples Taken with
High Volume Aerosol Sampling Runs 76
12. Microbiological Concentrations in Period 5 Composite and Characterization Aeration
Basin Samples 77
13. Enterovirus Concentrations in Period 5 Composite and Characterization Aeration Basin Samples 78
14. Trace Metals Concentrations in Period 4 Second Stage Aeration Basin Composite Samples 78
15. Trace Metals Concentrations in Period 5 First Stage Aeration Basin Composite Samples 79
16. Meteorological Summary for Period 1 and 2 High Volume Particulate and Andersen Air Samples 80
17. Meteorological Summary for Period 4 and 5 High Volume Particulate Samples 81
18. Meteorological Summary for Period 4 and 5 High Volume and Andersen Sampling Runs 82
19. Pathogen Isolations from Period 1,2 and 4 Andersen Air Samples 85
20. Microbiological Concentrations in Period 4 High Volume Aerosol Sampling Runs 87
ix
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TABLES (Cont'd)
Table Page
21. Microbiological Concentrations in Period 5 High Volume Aerosol Sampling Runs 88
22. Aerosol Particle Size Distribution for Standard Plate Count and Total Coliform 92
23. Summary of Source and Distance Analyses Performed on Period 4 and 5 Air Sample
Microbiological Data 93
24. Trace Metal Concentrations in Period 1 and 2 Paniculate Air Samples 96
25. Trace Metal Concentrations in Period 4 and 5 Paniculate Air Samples 97
26. Standard Plate Count for Soil Samples 104
27. ANOVA on Standard Plate Count in Soil Samples 104
28. Equality of Variance Tests—Ordinary vs Transformed 106
29. Determined Concentrations of Lead in Soil Samples 107
30. ANOVA on Lead in Soil Samples 107
31. Determined Concentrations of Copper in Soil Samples 108
32. ANOVA on Copper in Soil Samples 108
33. Determined Concentrations of Cadmium in Soil Samples 109
34. ANOVA on Cadmium in Soil Samples 110
35. Determined Concentrations of Mercury in Soil Samples 110
36. ANOVA on Mercury in Soil Samples Ill
37. Determined Concentrations of Zinc in Soil Samples Ill
38. ANOVA on Zinc in Soil Samples 112
39. Statistical Findings Summary—Soil Samples 112
40. Total Coliform for Water Samples 115
41. Standard PlateCount for Water Samples 115
42. ANOVA on Standard Plate Count in Water Samples 115
43. Lead Content of Water Samples 116
44. ANOVA on Lead in Water Samples 117
45. Copper Content in Water Samples 117
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TABLES (Cont'd)
Table , Page
46. Cadmium Content in Water Samples. :...:...:. : 118
47. Mercury Content of Water Samples. 118
48. ANOVA on Mercury in Water Samples : 119
49. Zinc Content of Water Samples 119
50. Statistical Findings Summary—Water Samples ; 120
51. Comparative Educational Levels 121
52. Comparative Occupational Distributions 121
53. Comparative Size of Residences , 121
54. Distribution of Households by Sector 122
55. Comparative Family Size Distributions 122
56. Comparative Racial Distributions 123
57. Comparative Distances from Treatment Plant 123
58. Comparative Distances from Nearest Expressway 124
59. Comparative Distances from Nearest Multilane Highway 124
60. Comparative Distances from Nearest Industry 124
61. Comparative Years of Residency at Present Address '. 124
62. Comparative Years of Residency in the City 124
63. Comparative Household Air Conditioning Distributions. :...... . 125
64. Comparative Age and Sex Distributions 125
65. Comparative Age Group Frequencies by Distance 126
66. Comparative Age-Sex Frequency 127
67. Chi-Square Independence Test Summary—Socioeconomic Characteristics .•• 128
68. Chi-Square Independence Test Summary—Exposure Characteristics 128
69. Incidence of Illnesses, Diseases, and Symptoms Reported in the Household Health Surveys 132
70. Test Statistics for Significance of Responses 139
XI
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TABLES (Cont'd)
Table Page
71. Physician's Evaluation of Response Categories 140
72. Incidence by Distance and Direction from the Egan Plant for Those Diseases and Symptoms
with a Potentially Implicating Distance Pattern 144
73. Percentage Incidence of Influenza by Age and Sex Group at 0 to 2.0 km Distance 147
74. Percentage Incidence of Vomiting by Age and Sex Group at 0 to 2.0 km Distance 147
75. Percentage Incidence of Dysentery by Age and Sex Group at 0 to 2.0 km Distance '. 148
76. Percentage Incidence of General Weakness by Age and Sex Group at 0 to 2.0 km Distance 148
77. Percentage Incidence of Nausea by Age and Sex Group at 0 to 2.0 km Distance 149
78. Percentage Incidence of Skin Disease by Age and Sex Group at 0 to 2.0 km Distance 149
79. Percentage Incidence of Diarrhea by Age and Sex Group at 0 to 2.0 km Distance 150
80. Percentage Incidence of Pain in Chest on Deep Breathing by Age and Sex Group at 0 to
2.0 km Distance 150
81. Summary of Health Survey Analyses 151
82. Participants by Group and Sampling Period 152
83. Age and Sex Distribution of Human Subject Participants 153
84. Racial Distribution of Human Subject Participants 153
85. Hair Color Distribution of Human Subject Participants 154
86. Marital Status Distribution of Human Subject Participants : : 154
87. Occupational Status Distribution of Human Subject Participants 154
88. Occupation Distribution of Human Subject Participants 155
89. Employer Distribution of Human Subject Participants 155
90. Occupation of Heads of Households 156
91. Educational Level of Heads of Households 156
92. Frequency of Chronic Illnesses Among Human Participants 156
93. Participants by Distance and Direction from Egan Plant 157
94. Distance from Nearest Expressway for Human Subject Participants 157
XII
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TABLES (Cont'd)
Table Page
95. Distance from Nearest Multi-Lane Highway for Human Subject Participants 158
96. Age-Sex-Distance Distribution of Human Subject Participants 158
97. Years of Residency at Present Address for Human Subject Participants 158
98. Years of Residency in the City for Human Subject Participants.. . 159
99. Hours per Day More than Two Miles from Home for Human Subject Participants 159
100. Cigarette Smoking Among Human Subject Participants 159
101. Selected Participants—Additional Serological Tests 160
102. Characteristics of Participant Groups 161
103. Age and Sex Distributions of Participant Groups .....' 162
104. Potential Variables for Regression Equation • 165
105. Summary of Positive Bacterial Isolations in Feces Samples and Throat Swabs 167
106. Comparative Analysis of Bacterial Isolations in Feces Samples 168
107. Comparative Analysis of Bacterial Isolations in Throat Swabs '. 169
108. Summary and Interpretation of Regression Equations for Bacterial Isolations. . 171
109. Summary of Positive Parasite Isolations in Feces Samples 173
110. Comparative Analysis of Parasite Isolations in Feces Samples 174
111. Summary of Positive Viral Isolations in Feces Samples 175
112. Comparative Analysis of Viral Isolations in Feces Samples 175
113. Summary and Interpretation of Regression Equations for Viral Isolations 177
114. Comparative Viral Antibody Titer Distribution from Original Tests on All Human Subjects 179
115. Comparative Viral Antibody Titer Distribution from Additional Tests on 100 Human Subjects .. 180
116. Summary of Viral Antibody Titer Comparisons from Original Tests on All Human Subjects .... 182
117. Summary of Viral Antibody Titer Comparisons from Additional Tests on 100 Human Subjects .. 184
118. Summary and Interpretation of Regression Equations for Viral Antibody Tilers 190
119. Summary of Cadmium Concentrations in Blood 192
XIII
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TABLES (Cont'd)
Table
120. Summary of Cadmium Concentrations in Feces ...................................... 193
121. Summary of Cadmium Concentrations in Hair ....................................... 194
122. Summary of Cadmium Concentrations in Urine ... ................................... 195
123. Summary of Lead Concentrations in Blood .......................................... 196
124. Summary of Lead Concentrations in Feces .......................................... 197
125. Summary of Lead Concentrations in Hair ........................................... 198
126. Summary of Lead Concentrations in Urine .......................................... 199
127. Summary of Mercury Concentrations in Blood ....................................... 200
128. Summary of Mercury Concentrations in Feces .......... .............................. 201
129. Summary of Mercury Concentrations in Hair ........................................ 202
130. Summary of Mercury Concentrations in Urine ....................................... 203
131. Summary of Copper and Zinc Concentrations in Blood ................................. 204
132. Summary of Copper and Zinc Concentrations in Feces ......... '. ....................... 205
133. Summary of Copper and Zinc Concentrations in Hair .................................. 206
134. Summary of Copper and Zinc Concentrations in Urine ................................. 207
135. Summary of Hematocrits in Blood ................................................ 208
136. Clinical Specimen Trace Metal Concentration and Hematocrits Patterns .................... 209
137. Current Health Status Data Summary: Incidence of "Presently Receiving Medication for
Chronic Illnesses" [[[ 211
138. Current Health Status Data Summary: Incidence of "Diseases Diagnosed During Past Year" ---- 211
139. Current Health Status Data Summary: Incidence of "Symptoms Experienced During Past
Three Months" [[[ 213
140. Current Health Status Data Summary: Incidence of "Symptoms Experiencing at Present" ...... 214
141 . Current Health Status Statistical Analysis: Sign Test on Increases and Decreases in Incidence .... 215
142. Summary and Classification of Findings Regarding Health Implications of a Sewage
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TABLES (Cont'd)
Table . Page
A-l. Aerosol Run Report on Meteorology and Sampling Configuration 273
A-2. UTSA-CART Aerosol Run Analysis Report 276
A-3. Household Health Survey 278
A-4. Health Survey Participant Questionnaire 280
A-5. Current Health Status Questionnaire 282
A-6. Metal Analysis Coding Form 284
A-7. Pathogen Analysis Coding Form 285
A-8. Pathogen Analysis Data Form 286
A-9. Bacterial Isolations in Feces Samples From Individual Participants 290
A-10. Bacterial Isolations in Throat Swabs from Individual Participants 296
A-l 1. Parasite Isolations in Feces Samples from Individual Participants 302
A-12. Viral Isolations in Feces Samples and Throat Swabs from Individual Participants 308
A-13. Coxsackievirus and Poliovirus Antibody Tilers from All Individual Participants 312
A-14. Echovirus Antibody Tilers by Hemagglutinalion Inhibition from All Individual Participants. 318
A-l5. Adenovirus Antibody Titers by Serum Neutralization from 100 Individual Participants .... 324
A-16. Coxsackievirus Antibody Titers from 100 Individual Participants 326
A-17. Echovirus Antibody Titers by Serum Neutralization from 100 Individual Participants 328
A-18. Echovirus Antibody Titers by Hemagglutination Inhibition from 100 Individual Participants. 330
A-19. Reovirus Antibody Titers by Hemagglutination Inhibition from 100 Individual Participants. . 332
A-20. Trace Metal Concentrations in Blood Samples from Individual Participants (/tg/100 ml) ... 334
A-21. Trace Metal Concentrations in Feces Samples from Individual Participants (/*g/g) . 340
A-22. Trace Metal Concentrations in Hair Samples from Individual Participants (/tg/g) 347
A-23. Trace Metal Concentrations in Urine Samples from Individual Participants (/tg/1) 352
A-24. Hematocrit Value of Blood Samples from Individual Participants 357
xv
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SECTION 1
INTRODUCTION
MOTIVATION AND PURPOSE OF THE STUDY
The United States Environmental Protection Agency (USEPA/EPA), through its construction grants
program to the states and municipalities, is funding a multi-billion dollar effort to construct new wastewater
treatment facilities throughout the United States. These new facilities are required to meet federal regulations
regarding discharge of wastes into rivers, lakes and oceans. The EPA also provides technical assistance to the
grant recipient regarding the design and location of these wastewater treatment facilities.
In the past, many wastewater treatment plants have been constructed away from populated areas, thus
avoiding problems with offensive odors in residential areas. When operated properly, modern wastewater
treatment facilities have fewer odor problems, so there is more acceptance of these facilities in residential
areas. From an engineering standpoint, the facility should be located as close as possible to the residential
area it is serving. Consequently, urban development has placed large populations in closer proximity to treat-
ment facilities in recent years.
Locating a wastewater treatment plant near residential areas may pose a possible health problem in that
sewage treatment processes generate aerosols which may contain human pathogenic microorganisms. A com-
monly utilized wastewater treatment technique is the activated sludge treatment process, which requires that
the incoming wastewater be aerated in biological treatment. During aeration, aerosols are formed at the sur-
face of the wastewater. The wastewater at this stage of treatment contains numerous bacteria and viruses
because little treatment other than removal of large solids has occurred.
As plans for this study, Health Implications of Sewage Treatment Facilities, were being formulated in
early 1974, the literature relevant to the microbiological aerosol health hazard was based primarily on en-
vironmental monitoring field studies of limited scope. Mickey and Reist*1) provide a useful summary of this
literature and evaluate its significance. The literature has adequately demonstrated that activated sludge treat-
ment processes are a source of microorganisms*2'7) which are carried at least 50 meters downwind from the
aeration basin/3-4'8) These studies used relatively low volume aerosol samplers, such as the Andersen six-stage
impactor, which lacked the sensitivity to distinguish between downwind and background concentrations
beyond 50 meters downwind from the source/3'4-6'7'8)
While direct evidence regarding health effects from the aerosols was scanty, the wastewater aerosols were
shown to be a source of pathogenic bacteria such as Klebsiella, Salmonella, Proteus, and Pseudomonas, that
might infect nearby residents/5-6'8'10'11) The presence and survival of tubercle bacilli in liquid sewage were
taken to indicate a potential tuberculosis health hazard/12) Enteric viruses, while not detected in wastewater
aerosol, were also considered to pose a public health hazard because of their prevalence in wastewater and
postulated survival in aerosols/11'13) The only reported health survey/14) concerning pneumonia incidence
among wastewater treatment plant employees, found these workers had a higher incidence of influenza
(+ 50%) and colds (+ 28%) than the control group of drinking water treatment workers.
There was a concern that activated sludge treatment plants might be a source of toxic metals such as cad-
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mium, lead, mercury, copper, and zinc. It was reported*15) that elevated levels of cadmium, copper, and zinc
were found in sewage containing industrial effluent. The toxic metals were presumably also present in the
aerosols from the aeration basins.
In summary, prior studies indicated that aerosols of microorganisms and trace metals were emitted from
wastewater treatment facilities such as trickling filters and the aeration basins of activated sludge processes.
These studies indicated that microbiological indicator measurements, such as standard plate count and total
coliform, and trace metals were elevated in the air just downwind of the wastewater treatment processes. Very
little information was obtained regarding specific types and concentrations of pathogenic microorganisms
present in the wastewater or in the air. Virtually nothing was known about the possible health effects on
populations living near these operations.
The purpose of the environmental epidemiology study reported herein was to identify possible health
effects which might be attributed to the operation of a wastewater treatment plant. The identification pro-
gram was to involve three independent modes of investigation of a selected new activated sludge treatment
plant, both before and after its initial operation:
• Environmental monitoring in the vicinity of the site to ascertain the source and transport distances
of indicator and pathogenic microorganisms and of trace metals;
• A health survey of a substantial cross-section of the households located within a 5-kilometer (3-
mile) radius of the plant aeration basin to examine the incidence of respiratory and intestinal diseases and
symptoms;
• Analysis of clinical specimens from over 200 participants residing within 3.5-kilometers (2-miles) of
the plant site for pathogenic bacteria, viruses, and parasites; to determine viral antibody liters; and to
measure trace metal concentrations.
Each investigation was to ascertain whether the observed data followed a pattern that implicated the
sewage treatment plant as a health hazard.
This study was intended to develop information on the health implications of sewage treatment to assist
the EPA in balancing the engineering and health concerns regarding locating wastewater treatment facilities
near populated areas, and to provide background information for making judgments on measures to
minimize aerosol formation. The EPA recognizes that a concomitant of funding is providing assistance in
plant design and siting to achieve cost effective wastewater treatment while minimizing the health risks to
nearby residents. Through the epidemiological research program it has initiated, which includes this study,
the EPA can develop the necessary design and siting information.
DESIGN OF THE STUDY
This study's design characteristics were developed to identify the health implications of operating sewage
treatment facilities by examining a new activated sludge treatment plant. The experimental design considera-
tions included determining the health hazards to be evaluated; constructing a sensitive design structure;
choosing a suitable study site; selecting the types of samples to be obtained in the three investigation areas (en-
vironmental monitoring, health surveys, and clinical specimens); specifying a suitable matrix of analyses of
sample types and sampling periods; and incorporating feasible state-of-the-art advances into the initial
experimental design.
Both microbiological and trace metal health hazards were selected for investigation. The microbiological
agents of concern were the enteric and respiratory human pathogenic bacteria and viruses and the parasitic
worms and protozoa. These agents were sought directly in the environmental samples and clinical specimens;
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and indirectly through indicator organisms in the environmental monitoring; through disease and symptom
incidence in the health surveys; and through viral serology of the blood specimens. The trace metals examined
were cadmium, copper, lead, mercury, and zinc. Their concentrations were measured in the environmental
samples and clinical specimens, and their effects were sought through the health survey symptomatology.
The general design structure was to make baseline (pre-operational) period versus operational period
comparisons at a new activated sludge plant in each of two seasons on the same participants and sampling
areas. The self-paired comparison of the results from the operational period against those from the cor-
responding baseline period is a very sensitive procedure for detecting changes because it eliminates the
substantial inherent variability between human subjects and between locations. The identified operational
period changes may be caused by operation of the sewage treatment plant.
Factors of longitudinal origin (changes over time) will be confounded with factors of plant origin in a
pre-post design. Supplementary analyses that separate plant and longitudinal factors in the operational period
are necessary to implicate the plant as a health hazard. The planned design structure made provision for these
supplementary analyses by specifying that measurement be taken over the detectable range of distances and
directions from the plant.
It is also important to the general design to select a new sewage treatment site. Sorber, et al/11) suggested
that treatment plant workers and nearby residents of an older plant might not show health effects from
exposure to microbiological aerosols, because sporadic inhalation of low concentrations of pathogens may
confer a degree of immunity/16) For an epidemiological investigation of microbiological hazards to have the
power to identify any health hazards that are present, newly exposed human subjects might be necessary.
With a new sewage treatment site, all the potential participants are newly exposed.
The following stringent site selection criteria were required: new activated sludge treatment plant that
would initially process at least 1,000,000 gallons per day of municipal wastewater; no existing sewage treat-
ment facility within at least a five-kilometer radius; a plant completion date consistent with the study
schedule; and a sufficient residential population within the five-kilometer radius, so the recruitment for an
adequate epidemiology study could be performed.
Though there are many new treatment plants under construction in the United States, most are additions
to existing facilities or new facilities being constructed adjacent to older ones. Another difficulty is that when
new plants are being constructed in new or expanding residential areas, they are ordinarily located as far from
the development as possible. After a diligent search, the John E. Egan Water Reclamation Plant — located in
the suburbs northwest of Chicago, Illinois — was selected. It met most of the site criteria, including about
40,000 people living within the five-kilometer radius. However, as the study commenced, only approximately
100 residences existed within a one-mile radius of the plant site, although numerous homes were under con-
struction within this inner circle.
Periods of several weeks duration in each of two seasons, fall and winter, were chosen for conducting the
full range of sampling in both the baseline and operational years. An early fall (late September-early October)
sampling period was chosen because the temperature in Chicago was still high enough to examine the poten-
tial enteric microbiological hazard, and because a return to the normal daily life-style pattern was presumed,
since the children had returned to school. A mid-winter (February) sampling period was selected to maximize
the potential respiratory microbiological hazard. Each type of sampling in the operational periods was con-
ducted, as nearly as possible, during the same week of the corresponding baseline sampling to minimize the
confounding from temporal and temperature-related factors.
The original human subjects selected as participants were not to be replaced if they dropped out of the
study. Each participant was to be his own control, so very sensitive self-paired comparisons of the correspon-
ding operational and baseline results could be performed. In the health surveys and for the soil and water
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environmental samples, the same locations were to be used in each sampling period, again to improve the sen-
sitivity of the comparisons.
Appropriate types of samples were to be taken in the three areas of investigation. The environmental
monitoring was to sample the ambient area, surface soil, and surface water in the vicinity of the sewage treat-
ment plant aeration basin, outward to residential distances (0.3 to 5 km) in all four sampling periods. In the
operational periods, the mixed liquor being aerated in the aeration basins was also to be sampled. The health
surveys were designed to obtain limited personal information on households and detailed background
information on volunteer participants. The household health survey was also to obtain disease and symptom
incidence on the household members through a door-to-door interview. Half of the households were to reside
within 3.5 kilometers in all directions from the plant site, with the other half residing between 3.5 and 5
kilometers in all directions from the plant. Suitable clinical specimens were to be obtained from the par-
ticipating human subjects for microbiological and trace metal analyses. The clinical specimens selected were
blood, feces, hair, throat swabs, sputum, and urine. A current health status survey eliciting the current
disease and symptom experience of the participants was also to be provided.
The study design specified the number of samples of each type to be taken in each of the four sampling
periods and the analytical procedures to be performed on each sample. The household health survey was to be
conducted only in the fall baseline and operational sampling periods, with health information being collected
on the members of 1000 randomly selected households in each survey. From 200 to 240 participants were to be
selected from the baseline household volunteers residing within the 3.5-kilometer radius, so at least 140 to 160
would remain to provide clinical specimens through all four sampling periods. The participants were to be
equally distributed among four desired age groups:
Pre-School 3 months to 4 years
School 6 to 12 years
. Young Adult 21 to 45 years
Elderly 60 years or older
The matrices of environmental samples and human subject samples actually collected and the types of
analyses actually performed are summarized in Table 1 for all of the sampling periods. The specific
microbiological organisms sought and assayed are given in Section 5, Analytical Methods, and Section 6,
Results, later in this report. The months during which the samples for each sampling period were actually
obtained are: . . ,.;
Period 1 September-October 1974
Period 2 February 1975
Period 3 September 1975
Period 4 February 1976
Period 5 September-October 1976
Improvements were incorporated in the original study design to take advantage of fortuitous cir-
cumstances and of fruitful avenues of investigation discovered in the related epidemiological studies. The
commencement of the Egan plant's operation was delayed over six months, so the plant was not yet operating
in the fall of 1975 when the fall operational sampling was scheduled to be performed. Meanwhile, the large
Lexington Green Apartment complex began accepting occupants just 400 meters to 700 meters northwest of
the plant aeration basin. Consequently, several design modifications were made.
The full fall operational sampling was postponed for one year. An interim clinical sampling was to be
substituted on approximately 50 additional participants recruited from the Lexington Green Apartments; this
interim baseline sampling period was called Period 3. The Lexington Green participants were to provide
clinical specimens in the fall operational sampling (Period 5) also. To conserve study funds, the analysis of the
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TABLE 1. ENVIRONMENTAL AND HUMAN SUBJECT SAMPLING MATRICES
Environmental Sample Matrix
Aeration Basin
Samples
Period: 1245
Number of Samples for
Microbiological Analysis 12 12
Bacteria X X
Viruses X X
Number of Samples for
Trace Metal Analysis 7 7
Cadmium X X
Copper
Lead X X
Mercury X X
Zinc
Human Subject Matrix
Blood Samples
Period: 1 2345
Number of Specimens Taken 171 170 34 192 216
Microbiological Analysis
Bacteria
Parasites
Viral Antibodies X X X X X
Viruses
Trace Metal Analysis
Cadmium X X
Copper X
Lead XX X
Mercury X X
Zinc X
Hematocrits XX X
Current Health Status Survey
Air Samples
1
28
X
X
28
X
X
X
X
1
211
X
X
X
X
X
X
X
X
245
27 32 72
XXX
XXX
28 33 31
XXX
X
XXX
X» X X
X
Feces Samples
234
206 45 185
XXX
XXX
XXX
1
50
X
X
50
X
X
X
X
X
5
218
X
X
X
X
X
X
Soil Samples
245
50 50 50
XXX
XXX
-
50 50 50
XXX
XXX
XXX
XXX
XXX
Hair Samples
1234
229
X
X
X
X
X
Water Samples
1 2
10 10
X X
X X
10 10
X X
X X
X X
X X
X X
5
190
X
X
X
4 5
10 10
X X
X X
10 10
X X
X X
XX
Sputum Samples Throat Swabs Urine Samples Health Questionnaires
1 234 5 1 234 5 12345 1 234 5
158 184 44 196 202 229 222 49 201 226 225 190 230 222 49 200 227
xxxxxxxxxx'
X X X X X
X X
X
X X
X X
X
X X X X X
X Samples taken and analyzed.
* Special mercury sampling for Periods 1 and 2 taken during April. 1975.
-------
winter baseline and operational period environmental samples and clinical specimens was deleted, since a
single seasonal comparison in the fall was deemed adequate to identify trace metal health hazards.
Our concurrent extensive aerosol and wastewater monitoring study of spray irrigation at Pleasanton,
California developed a superior microbiological aerosol sampling and analytical protocol. The aerosol
sampling complexities resolved at Pleasanton also implied that the microbiological air sampling design at the
Egan plant would be inadequate to accomplish this study's environmental monitoring study objectives. Con-
sequently, the same microbiological aerosol sampling and analysis protocol developed at Pleasanton was
utilized in the fall operational sampling in place of the original air sampling and microbiological analysis pro-
tocol.
A University of Cincinnati grant study demonstrated that viral serology tests for many virus types yielded
useful health effects measures. Therefore, 100 of our participants with sufficient blood sample volumes were
selected as three matched groups (1—Lexington Green, 2—close original participants, 3—far original par-
ticipants) and their fall baseline and operational blood samples were analyzed using 23 additional viral
serology tests.
RESEARCH QUESTIONS
The purpose of this study, to identify the health implications of sewage treatment facilities, has been for-
mulated as specific research questions that take into account the study design. The separate research questions
which have been identified for the three independent areas of investigation are listed below by the investiga-
tion area. The statistical analysis of the data resulting from each type of sample analysis has addressed the
relevant research questions to develop findings regarding identification of potential health hazards.
Environmental Monitoring
Determine whether the sewage treatment plant processes are a significant local source (just downwind of
the aeration basin) of pathogenic and indicator bacteria and viruses and trace metals in aerosol form.
If the plant is a significant local source, determine whether the levels downwind from the plant are
significantly elevated at residential distance.
Household Health Survey
Characterize the surveyed households by means of their personal, socio-economic, and exposure factor
distributions. Determine whether there are significant differences between the baseline and operational survey
distributions.
For those illnesses, diseases, and symptoms plausibly related to sewage treatment, determine whether the
frequency of occurrence in the operational year is significantly different than in the baseline year.
If the incidence is significantly higher, determine whether frequency pattern as a function of distance
from the plant implicates the sewage treatment plant as a potential health hazard.
Human Subject Sampling
Characterize the human subjects with respect both to their duration of participation and to their distribu-
tions of personal, socio-economic, medical history, and exposure factors.
For each pathogen analyzed, determine from the self- and seasonally-paired data whether occurrence in
the operational year is significantly more frequent than in the baseline year.
-------
If the pathogen incidence increases significantly, determine whether the increase relates to exposure to
the sewage treatment plant as opposed to other environmental and personal factors.
Determine whether human trace metal concentrations increased subsequent to plant operation.
Determine whether human subject health status declined subsequent to plant operation.
INTERPRETATION OF FINDINGS
The findings from the statistical analyses have been interpreted carefully. This has been an exploratory
study of the potential health hazards associated with sewage treatment plant operation. It has been fairly
comprehensive in the numbers and types of samples collected and analyses conducted. Because the study has
been both exploratory and comprehensive, it is to be expected that many findings will emerge in each area of
investigation and that most will be negative regarding sewage treatment health effects. It was also recognized
that isolated statistical findings are subject to error, both intentionally through the significance level set for
the hypothesis testing, and unintentionally through use of deficient data. Furthermore, while the study design
is good, longitudinal and sporadic effects can be confounded with the treatment plant effects in the findings
obtained.
Interpretation of the findings has been based on the criteria of consistency and rationality. The findings
in each area of investigation have been summarized. An apparent health hazard classification (negative,
uncertain, or positive) has been assigned to each finding by consensus of the investigators in the relevant
scientific disciplines. All the findings regarding a potential health hazard have been reviewed across all areas
of investigation. The potential enteric virus health hazard, for example, has been examined through the
findings from the aeration basin, air, soil, and water environmental sampling; from isolations in throat swab
and feces specimens and from antibody liters in blood sera; and from intestinal symptomatology in the health
surveys.
The consistency in classification and the reasonableness of the findings about a potential health hazard
have been considered, along with the likelihood for confounding, to determine whether it should be identified
as a sewage treatment health hazard. This process has led to our evaluation of the health implications of
sewage treatment facilities.
PARTICIPATING ORGANIZATIONS AND PRINCIPAL PERSONNEL
The research effort documented in this report has been conducted by Southwest Research Institute with
significant support and contributions of a number of other organizations. The following is a listing of par-
ticipating organizations:
—Southwest Research Institute (SwRI), San Antonio and Houston, Texas
—Southwest Foundation for Research and Education (SFRE), San Antonio, Texas
—The University of Texas at San Antonio (UTSA)
—Aqualab, Streamwood, Illinois
—Naval Biosciences Laboratory (NBL), Oakland, California
—Manpower, Incorporated, Chicago, Illinois
—John E. Egan Water Reclamation Plant, Metropolitan Sanitary District of Greater Chicago
(MSD), Schaumburg, Illinois
—Hanover Park Treatment (MSD), Hanover Park, Illinois
The study area extends outward from the John E. Egan Water Reclamation Plant to a radius of five
kilometers. Details regarding area of responsibility and principal participating personnel for organizations
other than the Metropolitan Sanitary District are presented in Table 2.
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TABLE 2. PARTICIPATING ORGANIZATIONS AND KEY PERSONNEL
Organization
Responsibility of
Organization
Key Personnel
Primary Responsibility
of Key Personnel
SwRI
SFRE
UTSA
Aqualab
NBL
Alexian Brothers
Medical Center
Manpower, Inc.
Project direction and final results
Analysis of clinical specimens, soil and
water samples for pathogenic parasites,
bacteria and viruses
Analysis of coliphage pathogenic
bacteria and viruses (wastewater and
aerosols, Period 5)
Air Pollution
Indicator microbial parameters
Consultant organization
Consultant organization
Temporary employment firm used to
hire field survey team
D. E. Johnson, Ph.D.
J. W. Register, Jr.
R. J. Prevbst
D. E. Camann
R. E. Thomas
J. M. Taylor
J. M. Hosenfeld
J. L. Gulinson
C. W. Hall.M.D.
S. S.Kalter.Ph.D.
C. H. Milstein
R. E. Kuntz, D.V.M.
J. Moore
B. P. Sagik.Ph.D.
C. A. Sorber, Ph.D.
M. N. Guentzel, Ph.D.
B. E. Moore
J.O. Ledbetter.Ph.D.
J.K. Hamilton
M. A. Chatigny
J. Clark, M.D.
T. Rooney
R. Purdie
D. Bennett
J. Leason
T. Suk
G.Tasker
Project Officer
Direction of field and lab
activities
Questionnaire Design,
Participant Recruitment
and Household Health
Survey
Statistician
Statistician
Air, water and soil
sampling
Clinical specimens
Air sampling
Meteorology
Medical Consultant
Direction of analysis
Pathogenic bacteria and
viruses
Parasites
Parasites
Direction'of analysis
Analytical methods
Virology
Air pollution consultant
Lab director
Consultants in aerobiology
Consultant in medical
matters
Air sampling
Air sampling
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The organization directing the study is Southwest Research Institute and all other organizations listed are
acting in support of SwRI in this study. All personnel listed are permanent professional staff at the various in-
stitutions, with the exception of those listed for Manpower, Inc., who are temporary employees hired for field
surveys. The field survey team exhibited the same high degree of professionalism as the permanent profes-
sional staff.
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SECTION 2
CONCLUSIONS
The John E. Egan plant appears to be a source of indicator bacteria, coliphage, pathogenic bacteria,
enteroviruses, and mercury in the aerosols emanating from its aeration basins. However, the levels of
microbiological or chemical agents of the air, soil, and water samples in the neighboring residential areas were
not distinguishable from the background levels.
From the patterns observed in the household health survey, the increased incidence of skin disease, and
the symptoms of nausea, vomiting, general weakness, diarrhea, and pain in chest on deep breathing may be
associated with the nearby operation of the wastewater treatment plant.
Results for alpha- and gamma-hemolytic streptococci isolations in the throat swabs for the subjects from
the Lexington Green Apartments provide some evidence that the pattern may relate to exposure to the
wastewater treatment plant aerosols. However, these results are of little practical health concern. In contrast,
31 viral antibody tests and attempted isolations of many pathogenic bacteria, parasites, and viruses yielded no
evidence of an adverse wastewater treatment plant effect.
The combined baseline—operational and distance experimental design used in this study is very sensitive
for identifying potential health hazards and inferring whether or not the wastewater treatment plant may be
their source. However, the findings obtained in this study, when considered overall, did not detect a public
health hazard for persons living beyond 400 meters from a well-operated wastewater treatment plant.
High volume aerosol samplers (1,000 liters/min. or higher for a minimum of 30 minutes) are needed to
monitor levels of bacterial pathogens in air near wastewater treatment facilities. Aerosol samplers which sam-
ple at one liter per minute do not provide the required sensitivity for this type of air monitoring.
Monitoring of viruses in ambient air requires much higher sampling rates or times than are needed for
bacterial pathogens because of the very low levels present.
10
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SECTIONS
RECOMMENDATIONS
Another more sensitive epidemiological study with persons living very close to a wastewater treatment
plant is necessary to investigate the public health hazard which might possibly be associated with the opera-
tion of such plants. The study should emphasize microbiological environmental monitoring, health diaries,
and viral serology. The majority of participants should be recruited from within 1.5 km of the plant. A
baseline and post-operational study of a large new sewage treatment plant is highly desirable.
The influent of the study plant should be screened to ascertain the most likely health hazards before
selecting the microbiological and chemical hazards to monitor in any future epidemiology study.
Primarily negative findings were found relative to adverse health effects related to the transport of
pathogenic aerosols to exposed populations. These results should not be accepted as conclusive findings.
Large amounts of public funds are being spent to improve the treatment of sewage in the United States. It is
very important that the EPA collect sufficient data (sanitary engineering, environmental, epidemiological) to
ensure that the design and operation of these facilities provide the necessary treatment of sewage and avoid
the production of adverse health effects in populations living nearby.
Environmental monitoring of wastewater and aerosols should be performed at several sewage treatment
plants located in various geographic areas of the United States. Much information can be obtained from these
types of surveys relative to effects that variation of factors such as meteorological conditions, wastewater
treatment procedures, and sewage composition have on levels of microbial pathogens in ambient air.
11
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SECTION 4
THE STUDY SITE
SITE SELECTION PROCESS
The design of this epidemiological study required that a sewage treatment plant be selected that had cer-
tain characteristics. However, before the characteristics and a plant site could be matched, it was necessary to
find out what plants were being built and expected to become operational between June and December of
1975.
The regional offices of the Environmental Protection Agency were contacted to determine if any
municipality or wastewater district had a new sewage treatment plant going into operation in the near future.
A review of the grant construction list by EPA personnel gave us further leads.
Another major source of information was the offices of the state governments. A phone call was placed
to the state capitol information office to determine what state agency would have information about new
sewage treatment plants in that state. Various state departments such as Health, Water Quality Board, or
Wastewater sections provided enough information to determine if follow-up calls were necessary to local
governments where the plant was under construction. Most of the 48 contiguous states were contacted in this
manner.
When a sewage treatment plant appeared to be a potential candidate for the study, detailed information
about the plant site and the surrounding area was obtained from the local government. In most cases, the city
engineer provided this information. Additional information was supplied by the mayor or city manager, or
the design engineer of the treatment plant.
Site Requirements
The selection of a plant site was based on a list of requirements supplied by the EPA. The list was broken
down into plant, population and meteorological requirements.
Requirements for Plant Selection-
The sewage treatment plant had to be a new plant being built on a new site and could neither be an expan-
sion of an existing plant nor built on the site of an old plant. The design capacity of the plant had to be one
million gallons per day (MOD) or larger and use the activated sludge method of treatment. The design of the
plant and the constraint of being a new plant eliminated a vast majority of potential sites. However, other
constraints also limited the choice of plant. The plant had to serve a residential area with no heavy industry
contributing to the waste influent, and there could not be another sewage treatment plant within a six-mile
radius.
Requirements for Population Selection-
The other major factor which influenced the selection or rejection of a plant was the population living
around the plant site. Within a 5-kilometer (3-mile) radius of the plant, a minimum of 1,000 households had
to be present. Ideally, a constant gradient of people was needed throughout the study area. This meant that an
12
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equal number of human participants could be selected at any radius of the circle and could be matched with
similar socioeconomic characteristics of other human participants anywhere in the study area. Thus, the site
selected should include households adjacent to the plant as well as at various distances from the plant in all
directions.
Meteorological Data-
Information relative to meteorological conditions near the prospective plant site was also considered.
Ideally if wind speed and direction were somewhat constant, then a large population living around the plant
could be exposed to the treatment plant.
Site Visit
Local Government-
The study area surrounding the John E. Egan sewage treatment plant is governed by six different
municipal governments located in two counties. Each of these municipalities was contacted and briefed on the
proposed activities of the survey team. Permits needed and any other constraints were cleared up before the
survey team began any activities in the area.
Without the excellent help and cooperation of the local governments, this study would have been diffi-
cult to complete. The village officers in Schaumburg, Elk Grove, Hoffman Estates, Roselle, Itasca, and Roll-
ing Meadows provided the survey team with maps and helpful information and assistance. Likewise, the
counties of Cook and DuPage provided valuable assistance.
Hospital Contact for Epidemiological Background of Area-
Background information concerning the epidemiology of the study area was obtained from James Clark,
M.D., of the Alexian Brothers Hospital. This hospital is located within the study area and serves a wide
population both in the study area and in surrounding communities.
THE SELECTED STUDY SITE
The John E. Egan Water Reclamation Plant
The selected study site was the Metropolitan Sanitary District of Greater Chicago's John E. Egan Water
Reclamation Plant (Sewage Treatment Plant—STP). The plant was placed in service December 16, 1975,
serving a design population of 160,000 people in an area of 49 square miles. A generalized flow diagram is
presented in Figure 1.
The wastewater treatment at the STP is primarily normal to low strength domestic-commercial waste
(personal communication). The area served has limited light industry. The influent biochemical oxygen
demand (BOD) concentration averages approximately 100 to 150 mg/1. The total suspended solids (TSS) con-
centration is in the same range (100 to 150 mg/1). The waste flow at plant start-up was in the range of
10-15 MGD.
The daily waste flow presently is in the range of 15 to 20 MGD. The monthly average daily flow is
approximately 17 MGD. The flow into the plant is somewhat equalized by a wet well and by allowing the col-
lection system (sewer lines) to act as holding devices. This is done in an attempt to distribute the daily flow
variations in a more uniform fashion. The design dry weather daily average flow is 30 MGD. At the present,
the plant is hydraulically underloaded /17)
The treatment plant can be operated in three different modes: as a conventional activated sludge plant; as
a step aeration and step loading plant; or as a contact stabilization plant. The plant is presently operated in
the conventional mode; that is, the raw sewage is discharged into the head end of a series of long tanks and
13
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Discharge
Disposal
Figure 1. John E. Egan Water Reclamation Plant Flow Diagram.
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aerated. The plant is a two-stage system as shown in Figure 1. The wastewater enters the first stage aeration
basin, is aerated, and then enters the first stage clarifiers where the solids are allowed to settle. Some portion
of the solids is returned to the aeration basin with the remainder discharged to the solids concentration tank.
The BOD and suspended solids concentration are each approximately 10 mg/1. Very little removal is ex-
perienced at the present time in the second stage reactor.
The effluent from the second stage clarifier is discharged to the multi-media filter consisting of coal and
sand. The effluent from the filter is chlorinated and discharged to the receiving water. The BOD and sus-
pended solids concentration at discharge are each approximately 3 mg/1. The effluent limits for the plant are:
BOD—4 mg/1
TSS—5 mg/1
NH3—summer, 2 mg/1
winter, 4 mg/1
The plant appears to be successfully meeting the effluent requirements at the present time.
The production of aerosols was the primary health concern investigated in this study. The first and sec-
ond stage aeration basins produce the greatest amount of aerosols as these are aerated by blowing diffused air
into the tanks. The grit chamber is aerated and the solids concentration tanks (thickener) have the capacity to
be aerated, but both potential sources are in enclosed buildings.
The John E. Egan plant is presently operating below its maximum design capacity from both a hydraulic
and organic loading standpoint, and is producing a high quality effluent.
The Study Area
Site Description-
A map of the study site is provided in Figure 2. The John E. Egan Sewage Treatment Plant is located at
the center of the map, on Salt Creek, a tributary of the Des Plaines River. The area is located in the northwest
portion of the Chicago Metropolitan area, approximately 35 miles from the downtown business district. A
number of suburban communities, incorporated as villages, surround the John E. Egan Plant. These are the
villages of Rolling Meadows, Arlington Heights, Elk Grove Village, Itasca, Schaumburg, and Hoffman
Estates.
The study site was divided into an inner and outer region, by concentric circles as shown in Figure 2, at
3.5 km and 5 km from the sewage treatment plant. Thus, the inner region lies within a circle of 3.5-km radius,
and the outer region is a bank from 3.5 to 5 km from the plant. Persons residing within the inner region served
the study as the population-at-risk and persons within the outer region served as a control group. For the pur-
pose of surveying, each of these regions is divided into sectors as depicted. The original design was to" have ap-
proximately equal population in each of twenty sectors, ten in each of the two regions. Each sector was to
define a specific direction and distance from the plant. Actually, the twenty-one sectors shown were
developed with use of maps and census data for the area.
To accomplish the study goal of 1,000 house-to-house surveys, fifty surveys would need to be accom-
plished in each of the original twenty sectors. Because participants were to be recruited from the surveys
within the inner sectors, eleven sectors were selected rather than ten. This would provide 550 household
surveys if 50 were accomplished within each sector, and would provide an adequate base for selection of the
250 participants for which the study was designed.
It will be noticed that the sectors have somewhat irregular boundaries. This was a result of "gerry-
mandering" the originally regular boundaries so a sufficient population was to be found in each sector. The
15
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Figure 2. John E. Egan STP project study area map.
16
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boundaries follow natural boundaries between neighborhoods where appropriate. After the initial design of
twenty-one sectors, a twenty-second sector was designated for a new community, Lexington Green, which
was developed very near the sewage treatment plant during the early stages of this study. Section 22 in-
corporates the portions of Sectors 1 and 11 within 1/2 km of the plant.
Characteristics of the Area-
The study area is located approximately 25 miles northwest of downtown Chicago, Illinois. The area
occupies a plain, which.for the most part, is only some tens of feet above Lake Michigan (579 feet above
mean sea level). Topography does not significantly.alter air flow, except that lesser frictional drag over Lake
Michigan causes winds to frequently be stronger along the lakeshore.
The study area is in a region of frequently changeable weather. The climate is predominately continental,
ranging from relatively warm in summer to relatively cold in winter. However, the continentality is partially
modified by Lake Michigan and to a lesser extent by the other Great Lakes. Temperatures of 96°F or higher
occur in about half the summers, while about half the winters have a minimum as low as-15 °F.
Precipitation falls mostly from air that has passed over the Gulf of Mexico with some influence from
Lake Michigan especially during the winter. About one-half the precipitation in winter and about 10 percent
of the yearly total precipitation falls as snow. Snowfall from month to month and year to year is greatly
variable. Freezing rain sometimes occurs but is usually light. Precipitation during the summer occurs as
thunderstorms which are often locally heavy and variable. Longer periods of continuous precipitation occur
mostly in autumn, winter, and spring.
The amount of sunshine is moderate in summer and quite low in winter. A considerable amount of
cloudiness, especially in winter, is locally produced by lake effect. Days in summer with no sunshine are rare.
The total sunshine in December, partly because of shorter days, is only a little over one-third the July total.
The quadrant from the east to about due north of the Egan treatment plant is a designated forest
preserve. Two four-lane highways (Higgins and Arlington Heights Roads) pass through portions of this
preserve. Estimated traffic patterns for each of these roads would be between 20,000 and 30,000 cars/day.
The remaining three quadrants of the study area can be described as a mixture of residential,
agricultural, and small business tracts. Residential areas are estimated to comprise about 30-40 percent of
these quadrants. However, rapid growth in the area is taking over the available farmland and would appear to
substantially increase the residential percentage within the next few years. At present, however, the
agriculture usage is predominately for grain crops such as corn, wheat, and oats. An estimated 10 percent of
the study area is presently commercial enterprise such as shopping malls and business zones along the major
thoroughfares.
Potential sources of metals or pathogens within the study area are not limited to any one section of the
area. Vehicular traffic would be the major contributor of metals to the environment. Interstate Highway 90
bisects the study area and the Northwest Tollway cuts across the northern section. These two multi-lane,
limited-access highways are two of the major roadways from the northwest suburbs to downtown Chicago
and as such are heavily traveled. Similar contributions that affect air quality from other multi- and single-lane
highways serving the needs of the 50,000 to 75,000 inhabitants of the study area would be expected. Also,
within the study area is a vast multi-level shopping mall complex with its many acres of parking that would
contribute to the air metal level. Flight paths to O'Hare International Airport cross the northeast, central,
and extreme southern portions of the study circle. The O'Hare Airport routinely has over 1,900 take-offs and
landings in each 24-hour period. Depending on the wind direction, flight paths over the study area would
change and the trace metals emitted from aircraft would affect the study area.
Pathogens might come from various sources both within and adjacent to the study area. Local farmers
17
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use a combination of chemical and biological fertilizers for their grain crops. As manure is spread by machine
over the fields some form of paniculate matter would be emitted to the atmosphere. No feedlots were observ-
ed within the study area. Adjacent to the study area are three sewage treatment plants approximately seven to
eight miles distance from the Egan plant. To the west and southwest two plants are located with treatment
capacities of four and one and one-half MOD, respectively. The other nearby plant is about seven miles to the
east in Des Plaines.
Characteristics of the Residents-
The information contained in the Chicago Standard Metropolitan Statistical Area (SMSA) book, based
on 1970 U.S. Census, was used to describe the residents of the area according to personal and socio-economic
characteristics. Those tracts which fell partially or completely within the 5-km radius around the site are com-
bined and summarized below.
The total population in the exposed tracts was 99,661, of which 49,985, or 50.2 percent were male and
49,676, or 49.8 percent female. The population was racially predominantly white (99.3 percent). From the
census data, a breakdown of the population in the area by age and sex categories is made which roughly cor-
responds to the breakdowns used in this report. The age groupings are: below 5, 5-19, 20-44, and 45 and over.
The numbers and percentages for each sex separately and for the total population of the surrounding area are
presented in Table 3.
The education level of the people in the area was fairly high. The median number of years completed was
approximately 12.5, which means that over half of the adult residents had taken some college work. Of the
total population, 7,140 or 7.2 percent, had completed at least 4 years of college work. The total persons
employed among these residents was 37,129, and of these, 11,625 or 31.3 percent, were classified as profes-
sional or managerial personnel.
The median income for the area based upon the 1970 census was approximately 6,000 dollars, and the
mean income was 6,300 dollars.
In the interim between the 1970 census and the survey periods of this study, certain changes occurred
within the area. The population of the area began to increase and a substantial rise was observed in home con-
struction. This consisted primarily of single family dwellings, but also included both townhouses and high-
rise apartments. The residents tended to be middle class with salaries in the range from $10- to $50,000 per
year, and a mixture of professional and blue-collar workers. The area is expected to continue this growth in
the coming years.
TABLE 3. AGE-SEX BREAKDOWN OF POPULATION SURROUNDING
JOHN EGAN PLANT (1970 Census)
Category
Male
Age Group
Less than 5
5-
20
45
19
-44
and over
No. %
6
17
18
6
,938
,971
,579
,298
13.9
36.1
37.3
12.7
Female
No. %
6
17
19
6
,300
,294
,935
,137
12.7
34.8
40.1
12.4
Total
No. %
13
35
38
,238
,265
,514
12,435
13
35
38
12
.3
.5
.7
.5
18
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SECTION 5
METHODS
HOUSEHOLD SURVEY AND PARTICIPANT RECRUITMENT
General Design
In this study, house-to-house surveys were used to obtain health information on households and to
recruit paid participants for more in-depth study. Surveys were accomplished before the initial operation of
the John Egan plant and again after operations were underway for a significant period of time. The general
design was to accomplish 1,000 house-to-house surveys before the initial operation of the plant, and to recruit
250 participants for an in-depth study to accomplish another 1,000 house-to-house surveys after plant opera-
tions were underway. Study participants were recruited during the initial house-to-house survey and these
participants remained throughout the study. The second house-to-house survey, accomplished after plant
operations, required no recruitment of participants.
Participants were recruited from the inner region of the study site (Sectors 1-11 in Figure 2 in Section 4).
An equal number of participants was desired from four separate age groups: 0-6, 7-18, 19-45, and 46+ years
of age. The only restrictions placed on participation were residence in the area for six months or more, an in-
tent to live in the area for at least two years following the initial contact, and no employment at the John Egan
plant.
Questionnaire Development
Three questionnaire forms were developed to serve the specific needs of the study. The first form was
used to directly obtain household health information for comparison of the general public health before and
after the start-up of plant operations and also to compare health of households living near the plant to those
living further away. Also, in this first form, a question was included which asked if a person would participate
as a paid volunteer in a health study. Thus, the form served as a recruitment device as well as a data collection
method. A second form was developed to collect pertinent background information on individuals who
indicated a willingness to participate in the study. A third form was developed to obtain more detailed health
status information on each participant at frequent intervals during the multi-year study period. Biological
samples were also obtained from each of the study participants recruited with the forms.
In the design of the questionnaire forms, project staff with experience in previous public health studies
involving recruitment and participation of individuals from the general public were assigned the task of pro-
ducing a preliminary design for each form. Inputs were obtained from Institute medical staff regarding health
and symptomatic information required for the forms. A preliminary design was accomplished and was
reviewed with staff of the sponsor organization. Revised versions of each of the three forms were then
prepared and submitted with appropriate backup materials to the Office of Management and Budget for
official OMB approval. These materials as submitted to OMB, are presented in Appendix A.
The questionnaire forms approved for use in this study are presented in Figures 3, 4, and 5. The
Household Health Survey Questionnaire, shown in Figure 3, served to gather information regarding
household health and to ask if a person wished to serve the study as a paid participant (Question 15). The
19
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Form Approved
OMB 1S8-RO117
STAFF USE ON
Cols 1-4
»*nn
LY
u
D
HOUSEHOLD HEALTH SURVEY
ExpiraiJuly 1977
2. Address: Street.
City_
Telephone
Coli 51-68
Cols 68-70
Coll 71-77
Col 80
3. How many persons reside in your household? .
Co* 5
D
4. For each person in your household, including yourself, please indicate the ege and sex beginning with the oldest end proceeding
to the youngest:
Coll 6-7 Coll 8-9 Coll 10-11 Coll 12 13 Coll 14-15 Cols 16-17 Coll 18-19 Coll 20-21 Cols 22-23
Permn i 23456789
mmmmmmmmm
Col 24 Col 25 Col 26 Col 27 Col 28 Col 29 Col 30 Col 31 Col 32
nnnnnnnnn
Col 32 Colt 24-32
Se»: 1-Male
2-Femele
5. Has any one of your household aver been diagnosed tn having any of the following chronic illneuei:
malignancies
asthma— hay (tver
diabetes
heart conditions
hypertension
chronic sinutitii & bronchitis
arthritis & rheumatism
rheumatic heart disease
thyroid disease (specify type)
liver disease (specify type)
kidney disease (specify typa)
(Person in Household)
t 23456780
6. During the past year, hat anyone of your household been diagnosed as having any of the following disc
polio
infectious jaundice
pneumonia
worms
skin diWBM
pleurisy
spinal meningitis
influenza
croup
sleeping sickness
anemia
dysentery
empyema
(Person in Household)
1 23456789
7. Has anyone in your household experienced any of the following symptoms during the past three months:
(Person in Household)
123466789
severe headache not relieved by aspirin
severe dizziness
severe pain in bones and joints with
high fever
severe weight loss
severe night sweats
hemorrhagic rash
canker sores around mouth
yellow eyeballs
sore throat
cough
cold
(continued)
Figure 3. Household health survey questionnaire.
20
Coll 40-49
Coll 50-59
Coll 60*9
Coll 70-79
Coll 10-19
Coll 20-29
Cob 3049
Coll 4049
Coll 6069
Coll 60*9
Coll 70-79
coii 10-19
Cols 20-29
Cols 3049
Cols 40-49
Coli6O*9
Coll 60*9
Cols 70-79
Cols 10-19
Coli2O-29
Cols 3049
Coll 4049
Cols 60*9
Coll 70-79 ColBO
Coll 10-19
Coll 20-29
Cols 3049
Cols4O49
Coll 60*9
Coll6O*9
Cols 70-79
Coli 10-19
"Cols 20-29
Cols 3049
Cob 4049
-------
(Person in Household)
34567
skin rash: tact:
arms & legs
- body
diarrhea
bloody diarrhea
bloody urine
burning on urination
nausea
vomiting
pain in chest on deep breathing
weakness o< arms o> legs
cough up blood
stilf neck with fever
stiff neck with rash
general weakness
draining ear
fever above 103'F
severe trouble with teeth
colicky pains in abdomen
brown urine
shortness of breath
mvutsii
head)
yellow skir
mess (not due to blow on
8. How long have you'
Lived in vour present citv or town? 1
Months
3-5 G-_8 9-1JI 12-23 24-35
2345 6
Cols 50 59
Cols 60-69 Col 80
Cols 70-79 [_7]
Cols 10-19
Cols 20-29
Colt 30-39
Cols 40-49
Cols 50-59
Cols 60-69 Col 80
Cols 70-79 [3]
Cols 10-19
Cots 20-29
Cots 30-39
Cols 40-49
Cols 50-59
Cols 60-69 Col 80
Cols 70-79 [9]
Cols 10-19
Cols 20-29
Cots 30-39
Coli 40-49
Cols 50-59
Cols 60-69
Cols 70-79 Col 80
[+1
D
Lived 91 your present address?
's have you and your family changed living quarters during the last five ye;
s your home air conditioned? 1-No
2~Y
-------
HEALTH SURVEY PARTICIPANT QUESTIONNAIRE
1. Name (in full)
2. Address: Street.
City _
Zip Code _
Telephone .
3. Date of Birth.
4. What is your sex?
S. What is your marital status?
1 -single
2-married
3-separated
-Day.
-Year
2-female
4-divorced
5-widowed
6. Which of these best describes your present occupational status:
1 -employed full time (including self-employed) 5-student
2-employed pert-time 6-ptay/nursery school
3-unemployed 7-pre-school
4-housewife 8-retired
7. What is your usual occupation? (please specify) -
8. If you are employed, what is the nature of the company for which you work? (please specify) -
9. How many hours of the day do you normally spend more than 2 miles from your home? .
10. What is the natural color of your hair? 1-brown 4-blond
2-black 5-gray
3-red
11. Have you ever smoked as many as five packs of cigarettes, that is, as many as 100 cigarettes during your
entire life? 1-yes 2-no
12. Do you now smoke cigarettes? 1-yes 2-no
13. How many years have you lived in your present city or town?
At your present addre«? yean
14. How many times have you changed living quarters during the last five years? _
15. Do you plan to move during the coming year? 1-yes 2-no
16. Have you ever been diagnosed as having any of the following chronic illnesses:
No
Form Approved
OMB 158-RO117
STAFF USE ON
Cols 1-4
»an
LV
n
D
_ hours
Yes
Presently receiving medication
(please specify type!
Tuberculosis
Malignancies
Asthma-Hay Fever
Figure 4. Health survey participant questionnaire.
ExplraUuly 1977
Coll' 5-30
Coll 31-SO
Cols 51-65
Cols 66-70
Cols 71-77
D
Col 80
Cols 5-10
Col 11
Col 12
Col 13
Cols 14-15
Cols 16-17
Cols 18-19
Col 20
Col 21
Col 22
Coll 23-26
Cols 27-28
Col 29
Cols 30-32
Cols 33-35
Cols 36-38
(continued)
22
-------
Presently receiving medication
(please specify type!
n,aty>t*>s
Heart f>nriitinnt
Arthrifk ft Rhp^m^ticm
Kirtney Hi space (sperify type!
17. Is your home air conditioned? 1-no 2-yes, window only 3-yes, central
1R What k the usual orrupation of the heart of your household? |ples«» specify) . .
19. What educational level has been completed by the head of your household:
Cols 39-41
Cots 42-44
Cols 45-47
Cols 48-50
Cols 51-53
Cols 57-59
Cols 60-62
Cols 63-65
Col 66
Cols 67-68
Col 69
2-8th grade 6-college completed
3-high school-incompleted 7-graduate school
4-high school-completed
YOU HAVE COMPLETED THE QUESTIONNAIRE
THANK YOU FOR YOUR COOPERATION
INTERVIEWER NOTE AND RECORD
20. Distance
21. Sector
22. Distance to nearest:
a. freeway, expressway or turnpike
b. other major muttilane traffic artery
c. large industrial operation
23. IMNOWO 123456
less than 2 blocks less than 1 mile
one mile or more
Col 70
Col 71
Col 72
Figure 4. (continued)
23
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STAFF USE ONLY
•Oton
CWtNCMT HCALTH VTATUB QUECTIONNAJM
n lor •*» 0* OM chronic •
3. Art you pratwitlv Ofcmg wiy othv P
1 -iranQuillnr
S-or*l
6-«n(ibioiici
4. During ttw p*ft VMT. !•»«• you b«*rt dUgnpwd n hewing »ny of Hit foUoiMine
polio
1 -V« 2-No
nrn«
D
D
D
D
D
D
D
D
D
D
D
D
Colt
Ool<
cm?
CMS
cut
Col 10
cm it
CM II
CM 13
cm 14
Col IS
Col 16
Col 17
cm eo
(continued)
Figure 5. Current health status questionnaire.
24
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5. H*v« you *tpciww«d my of ItM following fvmpIomi during lrt« pail tttrr* monlrii? At p
•* fwMjertt not rdtaffd by wprii
n diii-incM
« pi.n m bonei »nti joinli with high Uwr
h«morrri*9K rwh
______ m,
m<
i i I
— — ED'
— — m~-
— — m™
__ _ Q^,,.,,,
— — ED —
«.».»
j I I 001,35-26
C°""-M
mu.2930
I PI
I I I
— — on
Col>31>3?
Coll 33 34
Noodv OilrrhM
m
I I COH3.I-W
— on
— ED
Coll 41-42
Coll 43-44
" *^'" ^ I j j Coll 4S-46
. „ I—T~l c,,,,,,^
_ [ [ )
. | | | **«*
— — ED —
| | | C0..53-&
Coii 55 56
Coii 5
I L 1
drlining Mr
nrw ibovt 103*F
ED
ED —
Cod 61 -62
1 1 1
wv.il troutHe with IMth __ . [ | | g,,,, g-j^
— ED
— ED
colicky (Mint in •bdonwn Co|i ^^
— -. — En
— — ED
o»i6r«a
Col. 69-70
Cftli 71-72
t. ...I 1
uncnnntouinMi (not (to* to blow on b««d>
—. —on
Coii 73-74
Co. BO
Figure 5. (continued)
25
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Health Survey Participant Questionnaire, shown in Figure 4, served to obtain pertinent background informa-
tion on each participant. The Current Health Status Questionnaire, shown in Figure 5, served to obtain more
detailed health information on each participant at specific intervals during the study period.
Survey Design
Development of the specific survey design for the study required an initial site visit to survey the study
area. With the information gathered and contacts accomplished during the initial site visit, specific survey sec-
tors were defined and a survey team was recruited. Background survey materials were developed and the field
survey team was assembled and trained. These activities are described in the following sections.
Initial Site Visit-
An initial site visit was accomplished during August 1974 by the two project staff members responsible
for design of the household survey. The study area was canvassed and each neighborhood was examined
regarding the potential for performance of house-to-house surveys and recruitment of participants. Detailed
maps of the proposed survey areas were acquired and visits were made to the municipal and county govern-
ments under whose jurisdiction the survey would be accomplished to obtain permission for house-to-house
survey work. Accordingly, preliminary contacts were accomplished with each of the following to obtain
clearance for the surveys:
Cook County Sheriff's Office
DuPage County Sheriff's Office
Hoffman Estates Village Officials
Elk Grove Village Officials
Schaumburg Village Officials
Itasca Village Officials
Roselle Village Officials
Rolling Meadows Village Officials
A temporary manpower firm with offices in the area, Manpower, Inc., was contacted and arrangements
were made for an on-site coordinator. The local firm agreed to recruit survey field workers for the study and
arrangements were made for an initial training meeting with all field workers at a time immediately preceding
the first household survey in September 1974.
Preparations for the Survey-
With the information obtained during the initial site visit, the 21 specific survey sectors presented in
Figure 2 in Section 4 were defined.
A scheme was developed for payment of survey workers which included an incentive bonus for recruit-
ment of participants. With this scheme, each interviewer would be paid $3.00 per hour. The interviewers were
to be hired through the services of Manpower, Inc., for a fee of 50%. The effective cost to SwRI was to be
$4.50 per hour. It was estimated that 50 completed questionnaire forms could be collected in a 40-hour work
week by each interviewer. To accomplish the desired 1,050 surveys in a one-week period would require 21
interviewers at the estimated rate of 50 completed surveys per week. A bonus of $1,000 was established for an
incentive to the workers to recruit study participants. A $4.00 bonus per participant questionnaire form col-
lected would be paid to the survey worker for each participant up to the 250 participants desired. If more than
250 participants were attracted to the study, the $1,000 would be prorated accordingly.
Instruction sheets for the survey workers and a handout sheet were prepared for use during the survey.
Examples of these backup materials and a news release for dealing with the news media are presented in
Appendix B, Survey Backup Materials. Plans were made to pay each survey participant $12.50 for each of
26
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four sampling periods and a bonus of $25 for completing all four periods. Thus, each participant could
receive a total of $75. Because participants were desired only for inner sectors, the instruction sheets
developed for interviewers are labeled Participant Sectors and Outlying Sectors, respectively.
The Survey Team-
Twenty-one survey workers were recruited and trained, including one on-site coordinator, Mrs. Carol
MacArthur. Mrs. MacArthur remained active as the study on-site coordinator throughout the remainder of
the study and was instrumental in making arrangements for all of the survey and sampling periods.
Each survey worker was assigned a specific sector, and was instructed to perform 50 house-to-house
surveys within the sector during a one-week period. The workers were instructed regarding the appropriate at-
tire, manners, and procedures to follow in performing the survey.
Performance of Baseline, Additional Baseline and Operational Household Surveys
Arrangements were made with two newspapers in the Chicago area to provide public awareness regar-
ding the study and create a climate of understanding and public acceptance of the survey activities. Articles
were published during September 1974 in the Chicago Tribune and in the Arlington Heights Herald, a
newspaper specifically serving the study area. During subsequent survey periods in 1975 and 1976, the
Arlington Heights Herald again published articles regarding the study.
The baseline household surveys, accomplished in September 1974, were directed on site by a member of
the SwRI project staff and coordinated by Mrs. Carol MacArthur. One field survey team member was assign-
ed to each of the 21 study sectors and was instructed to obtain fifty completed Household Health Survey
Questionnaire forms at random within the assigned sector.
Using these procedures, 1,043 household surveys were accomplished by the field survey team. Of these
surveys, 521 households in the inner region were asked to participate in the individual health survey. The
response was extremely high, with 471 individuals volunteering to participate in the study. This is an excep-
tionally high rate of response to a.public health survey requiring delivery of biological samples.
From the 471 individuals volunteering for participation in the study, some 250 individuals were selected
as participants. The volunteers were divided by sex and age into four groups. In selecting the participants, an
attempt was made to have an equal number of participants in each of the four age groups, and an equal
number of each sex in .each age group. Table 4 shows, by sex, the number of participants selected in each of
the study age groups.
TABLE 4. INITIAL PARTICIPANTS RECRUITED IN
SEPTEMBER 1974
Age Group (yr)
0-6
7-18
19-45
46+
Total
Number of
Male
22
33
40
15
110
Participants
Female
28
25
52
16
121
27
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The scheduled start-up of the plant was delayed past the September 1975 date of the originally scheduled
operational period survey. Also, during the intervening period of time, a new community of residents was
established very near the plant site, in the portions of Sectors 1 and 11 within 1/2 km of the plant. This is the
community of Lexington Green and was designated Sector 22. Because of the nearness of these residents to
the plant site, they were viewed as superb subjects for study participants.
The delay in the start-up date of the plant operations provided a possibility for baseline household
surveys to be conducted in September 1975 in the new area and recruitment of an additional set of study par-
ticipants. Accordingly, in September 1975 approximately 75 household surveys were accomplished in the Lex-
ington Green area and more than 40 new participants were recruited for the study. These participants were
added to the complement of those recruited during the initial baseline survey, and the combined group of par-
ticipants was studied from that time forward. The number of additional participants obtained in each of the
study age groups is shown, by sex, in Table 5.
TABLE 5. ADDITIONAL PARTICIPANTS RECRUITED IN
SEPTEMBER 1975
Number of Participants
Age Group (yr)
0-6
7-18
1945
46+
Total
Male
1
1
13
3
18
Female
7
4
15
7
33
In September 1976, the operational period survey was accomplished in the 22 study sectors with 1,104
household surveys . No participants were recruited during the operational period survey, as only participants
from earlier phases were required. Accordingly, the Household Health Survey Questionnaire form was
modified so not to include a question regarding participant recruitment.
The September 1976 household survey was arranged almost wholly through the work of the on-site coor-
dinator, Mrs. Carol MacArthur. Members of the project staff worked closely with Mrs. MacArthur to coor-
dinate her activities. Contacts were again made with all pertinent local governments and clearance was
established for the required survey activities. With the help of local temporary manpower firms, Mrs.
MacArthur recruited and trained 22 survey workers for use in the survey and supervised their activities during
the performance of the survey.
ENVIRONMENTAL SAMPLING
Organization Overview
The collection of environmental samples such as soil, air, water, and sewage effluent over the rather large
study area (6-kilometer diameter circle) required notification and obtaining permission from the following
municipalities:
—Schaumburg, Illinois
—Elk Grove Village, Illinois
—Roselle, Illinois
28
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—Hoffman Estates, Illinois
—Itasca Village, Illinois
—Rolling Meadows, Illinois
—Cook County, Illinois
—DuPage County, Illinois
—Metropolitan Sanitary District of Greater
Chicago (MSD)
—Forest Preserve District of Cook County,
Illinois
When required, formal permit agreements, certificates of insurance, and release of liability statements
were entered into or obtained from the municipality, governmental agency, or private citizens.
In the case of air sampling, when electric power was required, private residences were utilized whenever
wind conditions permitted. The owner of the residence was paid fifteen dollars for access to his property and
for the use of electric power. When no permanent source of power was available, gasoline-powered
generators were used.
Because of the diversity of tasks to be accomplished, two on-site managers, microbiological and heavy
metals, were designated to direct the activities of the special collection teams.
The microbiological manager had the overall responsibility for the field operation. His special duties
were to direct the activities of the microbiological collection teams in the collection and processing of the soil,
air, and water samples to be used for bacteria and virus determinations. In addition, he established daily sites
for the air samplers, taking into consideration wind and weather conditions for that particular day.
In some cases, it was more cost effective for the two collection teams to collect samples simultaneously at
the same site. In such cases close cooperation between the two groups was effected. The microbiological col-
lection team composed of up to six individuals were given extensive training for up to three days on the proper
procedures and care required in air sampling for bacteria and viruses. This instruction was given by a profes-
sional microbiologist. These people were all hired through a temporary employment agency and all but one
were either undergraduates or graduates in the field of science.
The heavy metals manager had the responsibility for directing the personnel involved in the air, water,
and soil sampling for heavy metal determinations. The heavy metal sample collection team was composed of
two members with one a professional chemist from SwRI and the other a person hired on site through the
temporary employment agency. Training for this job was usually accomplished during the first day of
sampling.
Air Samples
Sampling Design and Protocol-
In sampling periods 1 and 2, four portable air sampling stations were used to take 30-minute to 24-hour
samples along the wind's direction on each of the seven sampling days from October 13-19, 1974 and January
26-February 1, 1975. These samplers were located at any four of the five upwind and five downwind distances
from the plant (5, 50, 500, 1600, or 5000 meters). The sampler location design shown in Figure 6 was used
depending on the daily predicted predominant wind direction.
During the first two sampling periods a comprehensive monitoring program was conducted for seven
consecutive days beginning on a Sunday preceding the week the subjects were sampled. These air samples
were monitored for trace metals and bacterial and viral pathogens. All air samples were taken with regard to
wind direction and distance from the water reclamation plant. Five distances were used to describe the pattern
of the materials from this plant (5, 50, 500, 1600, and 5000 meters).
29
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Predicted
Wind Direction
East (Crossing
190 within 1 mile
of site)
North/West/South
North/West/South
North/West/South
North/West/South
Pattern
Upwind Downwind
Air-Sam pier Air-Sampler
Locations Locations
UOUT, U-IN
U-IN
U-IN
50, 1600
5, 50, 500, 1600
50, 500, 1600, 5000
5, 50, 500
500, 1600, 5000
Maximum
Pattern
Frequency
1
2
2
1 or 2
1
The location distance codes in the chart are relative to the center of the
aeration basin. They should be given the following interpretation:
UOUT:
U-IN:
5:
50:
500:
1600:
5000:
Upwind-out. Approximately 1600 meters upwind.
Upwind-in. About 200-800 meters upwind.
Downwind 5 meters.
Downwind 50 meters.
Approximately 500 meters downwind.
Approximately 1600 meters downwind.
Approximately 5000 meters downwind.
Figure 6. Air sampler location design.
30
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Sampling points along these circles were selected with regard to direction. Four air samplers were used
for collection of ambient air samples and one of the air samplers was always located on the upwind side with
the other three on the downwind side. The upwind station was intended to describe the background levels of
microorganisms and trace metals.
For sampling off the plant grounds, 24 sampling sites were preselected to describe the windrose pattern at
1.6- and 5-kilometer distances from the plant. These sites were located based on the windrose patterns for the
time of year being sampled. In addition, 12 additional sites were preselected in case the wind direction was dif-
ferent than that predicted by the national windrose pattern charts for the area. The sampling sites were
changed each day to follow the wind direction. If the wind direction changed drastically during any one
sampling period, attempts were made to move the samplers.
The actual distances and the sampling direction each day were recorded in a log book to allow a more
accurate statistical analysis of the air sample data. If the sampling patterns were repeated on two different
days, attempts were made to sample along two different wind directions on those days. Hence, the actual air
sampling schedule followed on a given day was not assigned until the day before when a reliable wind direc-
tion for that day could be forecasted.
Expanded aerosol sampling was recommended for sampling period 5 operational environmental survey
primarily because of the importance of the findings in the period 4 sampling. Preliminary data obtained dur-
ing the winter operational survey indicated that the levels of some bacteria and viruses near the plant might be
as high or higher than levels seen near the spray irrigation facility in Pleasanton, California. The design and
protocol for sampling in periods 4 and 5 were the same except that in period 4 only one high-volume
microbiological sampler was available, while in period 5, simultaneous sampling at six stations was
accomplished.
Increased aerosol sampling was recommended because of the improved methods of sampling and
analysis that were developed in the recently completed Pleasanton environmental sampling program. The
Pleasanton study indicated that several high-volume samplers operating simultaneously were much more
effective in describing the levels of microbiological aerosols. The availability of these high-volume samplers
(LEAP and Litton) also made the proposed expansion of Chicago aerosol sampling feasible.
A second reason for recommending the expansion of aerosol monitoring was that the project scope of
work called for two operational samples to be collected around the Egan plant; one during the winter, and
one during the summer. However, the John E. Egan plant was not in full operation at the time of the winter
sampling and it was felt that a thorough summer aerosol sampling program would provide much essential
biological aerosol information that was lacking.
Basically, the proposed plan for expanded aerosol sampling during the summer operational survey was to
perform ten aerosol sampling runs over a seven-day period using six high-volume samplers (LEAP and Litton
type) operating simultaneously, with one upwind and five downwind. This basic design of sampling was
similar to that of the study near Pleasanton, California, and the recommended analytical protocol for both
aerosol and effluent samples was identical. This was an increase.over what had been utilized in previous
surveys at the John E. Egan plant and represented an increase in the scope of work of the study. In this pro-
posed expansion a total of 60 aerosol samples was collected. Each of these 60 aerosol samples was analyzed
for total plate count, total and fecal coliform, coliphage, pathogenic bacteria — salmonella, shigella,
pseudomonas, streptococcus, proteus, viruses — via three- and five-day counts — polio, adeno, coxsackie,
and echo.
A comparison was made of the aerosol sampling analyses results obtained at the Egan plant in the winter
sampling and at the Pleasanton spray irrigation site. From this comparison it was projected that, because of
diffusion and biological decay, the maximum feasible downwind sampler distance from the Egan plant at
31
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which statistically usable biological analysis results would be obtained was approximately 200m during the
day and 400m at night. It was recommended that these maximum distances replace the 5000m maximum
sampler distance of the existing aerosol sampling protocol. Because the simultaneous aerosol samplers were
located at varying distance intervals under the proposed plan, it was still possible to project the biological
aerosol concentrations out to 5000m using a suitably calibrated dispersion model. It should be noted that the
nearest residential areas to the Egan plant are now within 400m of the aeration basins.
Extra days were included above the actual seven days of sampling as a precaution for contingencies such
as rain, other environmental conditions, or equipment failure which might have prevented sampling or made
it necessary to abort sampling runs already initiated. Four runs were made in the early afternoon (Stability
Classes A-C with high solar radiation), three runs in the early morning (Stability Classes C-F with low solar
radiation), and three runs at night (Stability Classes C-F with no solar radiation).
The five downwind aerosol samplers were arranged along a sampler line determined by the mean wind
direction measured just prior to actual setup of the samplers. This compass direction would radiate out from
the center of the aeration basins. Sampler location distances from the edge of the aeration basin fell into the
following distance ranges:
Distance Ranges for Location of Downwind Samplers
(Meters)
15- 30
50- 75
100- 150
200- 400
600-1200
An upwind station was located directly upwind at a distance of 500 to 1600 meters for each run. Exact
distances for both upwind and downwind stations were recorded for each aerosol run.
Distance ranges were given due to the fact that exact distances were dictated by actual wind direction and
resulting interferences due to physical obstructions such as buildings, ponds, highways, trees, etc..
Duplicate samples were collected with samplers set up next to each other at the 15-30 meter distance dur-
ing one afternoon run and at the 50-75 meter distance during one night run. All other aerosol runs were con-
ducted using a single sampler at each of the five distance ranges mentioned above. In the event wind direction
and physical obstructions prevented placement of samplers at one of the distance ranges, the sampler in ques-
tion was placed at the nearest unobstructed point.
The procedure for field measurement of wind direction determines the mean wind direction, to the
nearest 5 °, from the field wind measurement strip chart recorder just before the aerosol sampling run.
To obtain the wind stability class in the field, one or more of the following procedures were followed
(listed in decreasing order of reliability):
—from the temperature difference instrumentation (4.7m vs 0.5m):
Reading Interpretation Stability Class
+ Stable E or F
0 Neutral D
— Unstable A, B or C
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—from the solar altitude, wind speed, and cloud cover as described by D. B. Turner in Atmospheric
Dispersion Estimates (Table 6).
TABLE 6. KEY TO STABILITY CATEGORIES
Surface Wind
Speed (at 10 m),
msec'l
2
2-3
3-5
5-6
6
Day
Incoming solar radiation
Strong
A
A-B
B
C
C
Moderate
A-B
B
B-C
C-D
D
Slight
B
C
D
D
D
Night
Thinly Overcast or
> 4/8 Low Cloud
E
D
D
D
<3/8
Cloud
F
E
D
D
The neutral class, D, should be assumed for overcast conditions during day or night.
—from the variability in wind direction as described in the following table:
Allowable Variations in Wind Direction for
Stability Classes
Allowable Standard Deviation
About the Mean Wind Direction
Stability Class (degrees)
A
B
C
D
E
F
± 25
± 20
± 15
± 10
± 5
± 2.5
A sampling run was not set up if rain or other environmental conditions existed that would interfere or
affect the authenticity of the sampling run.
If it was decided to sample, the aerosol run was set up and the wind direction and stability were recheck-
ed. Sampling occurred only if the wind direction mean remained fairly constant and if no marked changes in
meteorological conditions were expected over the next 30 minutes.
The run was continued as long as there was no marked change in the mean wind direction prior to the
completion of 15 minutes of sampling. If during the first 15 minutes the mean wind direction shifted more
than 45 °. from the direction at the beginning of the run, then the samplers were no longer downwind of the
spray line and the run was aborted.
The aerosol samples were processed and shipped for analysis unless one or more of the following condi-
tions occurred:
—Based on the field strip chart recorder and temperature difference instrumentation, there was
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sufficient change in the wind direction and/or stability that three or more samplers experienced
less than 15 minutes of actual downwind sampling.
—Aerosol samples were obtained from less than three of the downwind samplers.
—No upwind aerosol sample was obtained. If the upwind (background) sampler should malfunc-
tion during the run, another upwind sample could have been substituted provided it was taken
within two hours after run completion and wind direction had not reversed.
Sample Collection Procedures for Metals and Microbiological Aerosol Samples-
The collection of air samples was performed to define metal concentrations (lead, zinc, copper, cad-
mium, and mercury) before and after the treatment plant went into operation.
The equipment used to collect the air samples consisted of high-volume paniculate samplers of standard
design. Equipment from three different manufacturers (Staplex, BGI, and General Metal Works) were used
on this study. Basically, each consisted of a blower with an 8" x 10" holder for the filter. A weatherproof
unit composed of wood or aluminum, depending on the manufacturer, protected the filter and the motor
from the weather. Prior to sampling operations in the field, each high-volume sampler was calibrated with a
series of resistance plates. A secondary calibration curve was then developed for each sampler based on the
primary calibration curve for the resistance plates.
Glass fiber filters, Type A (20.3 x 5.4 cm), without an organic binder (Gelman Instrument Co.), were
used for the paniculate collection media. A light table was used for visual inspection of each filter for
thickness variation and pinholes. Filters passing this test were then sequentially numbered and separately
placed in clean polyethylene bags. After sample collection, these filters were analyzed for lead, zinc, copper,
and cadmium levels.
Sample collection for mercury in air involved the use of three different methods during different phases
of the project work.
The charcoal tube method developed by Scarengelli(18) was used for all phases of this project except the
last post-operational period. For that period the silver wool absorption procedure developed by Long(1^ was
used. This method was also complemented by use of the potassium permanganate procedure of Gardner/20^
Five distances were used to describe the pattern of metals from the treatment plant — 5, 50, 500, 1600,
and 5000 meters. Sampling points were identified along these grid lines and where possible homeowners near
the identified sites were contacted. The placement of the samplers in the homeowner's yard provided security,
a source of electricity, and, most importantly, the close-hand monitoring of ambient air levels within the
neighborhood. Twenty-eight preselected sampling sites were used off the plant grounds and ranged from 500
to 5000 meters from the center of the primary aeration basin. Each site selected had the minimum amount
possible of obstructions that might alter the air flow pattern from the treatment plant.
Before the start of each day of sampling, the weather bureau at O'Hare Airport was called to find out the
prediction for wind direction during the following 24-hour period. Based on this prediction and the current
wind conditions at the plant site, four sampling sites were identified for that 24-hour period. One station was
located upwind and a minimum of three stations were located downwind. Generally, the stations located on
the plant site were set up first, followed by the stations in the surrounding community.
At each site, one high-volume and one mercury sampler were set up. A clean, numbered glass fiber filter
was placed in the high-volume sampler. Next, a precleaned, numbered mercury collection tube was connected
to a vacuum pump with a critical orifice. The high-volume sampler and the mercury pump were then started
and the exhaust from them was directed downwind. A 10-minute warm-up period was allowed for each
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sampling unit motor to come to operating conditions. Start-up time, flow rate, filter and tube numbers,
sampling unit identification, and wind conditions were recorded for each site. Approximately 12 hours later,
each sampling site was visited to ensure there were no problems with the samplers.
At the end of the 24-hour sampling, the flow rate for each sampler was noted as was the time the
samplers were turned off. The glass fiber filters were removed from the sampler and folded with the
particulates on the inside. The filter was then returned to its polyethylene bag and labeled appropriately. The
mercury collection apparatus was similarly sealed and labeled with site number, day, date, and unique sample
identification number. The samplers were then set up again either at the same site or moved depending on the
wind direction. This process of setting up and maintaining the samplers at the designated downwind and up-
wind distances continued for a total of seven consecutive sampling days for each sampling period. At the end
of the sampling period, the collected samples were sent to the San Antonio laboratories for analysis.
Samples were obtained for bacteria using both Andersen 2000 samplers and high-volume electrostatic
precipitators (LEAP and Litton). Samples for viruses were obtained using a one cfm impinger. The samplers
are described below.
The Andersen 2000 is a two-stage sampler designed to use commercially available standard 100-mm Petri
dishes pre-filled with 20 ml of sterile bacteriological agar. The sampler is designed to draw one cfm of air
when attached to a vacuum source. The following types of media were used in the Andersen 2000 samplers.
—blood agar
—MacConkey agar
—Lowenstein - Jensen
—Salmonella - Shigella
—Middlebrook7H10
Two types of large-volume air samplers were used for collecting biological aerosol samples. The first type
was the LEAP Sampler, Model 3440, manufactured by the Environmental Research Corporation of St. Paul,
Minnesota; the second type was the Litton Model M Large-Volume Air Sampler, manufactured by Applied
Science Division, Litton Systems, Inc., Minneapolis, Minnesota.
The two different types of samplers are designed to collect airborne particles and concentrate them into a
thin, moving film of liquid. The samplers are basically electrostatic precipitators. Aerosol is drawn into the
sampler through a converging nozzle and passes through the center of a high-voltage plate, then flows radially
between the plate and a lower rotating collection disc. A ring of needles concentric with the air inlet emits a
continuous corona which imparts an electrical charge to the particles. The electrical field then provides the
driving force to precipitate charged particles onto the lower disc.
The medium used in these samplers was BrainHeart Infusion broth (BHI) with 0.1-percent Tween 80.
This was selected based upon the results of a transfer media study conducted as a part of "Evaluation of
Health Effects Associated with the Application of Wastewater to Land"^21^ where similar samples were
obtained.
The following operating procedure was used in obtaining aerosol samples with the high-volume samplers
(LEAP and Litton):
1. After sampling site was selected, the sample table was set up and ensured to be level. The
generator was set off to one side to the full length of the extension cord and the gas and oil
levels were checked before starting.
2. All high-volume air samplers were cleaned in the field immediately before use by the
following procedure:
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—scrubbing the rotating plate with Alconox or Clorox solution using a Kimwipe;
—running Clorox solution through the lines and onto the ground;
—running thiosulfate solution through the lines and onto the ground;
—giving a final rinse by running sterile water through the lines; and
—during cleaning, the liquid pump and the air blower should both be on but the high voltage
power supply should be off.
3. After the sampler was cleaned, the BHI medium bottle was removed and the level of the
liquid marked. The medium was run into the high-volume sampler and when the medium
began to come out the return lines, the liquid level on the BHI bottle was marked again. The
liquid was kept at this second level throughout the run by adding sterile water as necessary.
4. The plate was ensured to be wetting properly and then the liquid flow rate was set at 12
ml/min. The air flow rate was adjusted to read 1,000 1pm. Samplers were checked for about
one minute prior to the start of the sample run with the high voltage supply on to ensure that
no arcing or power problems existed.
5. When the signal to start sampling was given the high voltage power supply was turned to
14kv and a 30-minute sample was taken.
6. Careful attention was paid to the level of the BHI and to excessive arcing.
7. At the end of 30 minutes, the high voltage power supply was turned off and the supply line
pulled out of the BHI reservoir. The liquid pump and the air blower were run until all the
liquid was collected in the original bottle. The level was made up to the original 100 ml mark
on the bottle with sterile water.
8. The cleanup procedure was repeated.
The aerosol particle-size sampling protocol was to perform two particle-size sampling runs during the
period from September 30 through October 8, 1976. Six-stage Andersen viable particle samplers were located
at one upwind and five downwind stations for simultaneous collection of total coliform organisms on one run
and for standard plate count on the second run. Distance ranges for location of the samplers were the same as
for high-volume samplers. Sampling times were determined by a preliminary run for which the sampling times
were one minute for samplers located out to 30 meters, five minutes for samplers located out to 75 meters and
15 minutes for the more distant stations. Plates from this run were incubated and counted only if all stations
or plates were not overgrown. If all plates were countable, the run was considered valid and the second sampl-
ing run was conducted using the same sampling times. If the majority of the plates were not countable, sampl-
ing times were altered as needed for the second run or until two valid runs were obtained.
Air samples for viruses were obtained using a one cfm impinger. Collection of viruses from air was set up
by using a side-arm flask with a one-hole stopper through which passed a short length of glass tubing with a
funnel on the upper end and a capillary tip on the lower end extending into a known volume of BHI medium.
A source to produce a vacuum was attached to the side-arm and the diameter of the capillary regulated the
flow of air which passed through the medium in which the viruses were collected. One cfm rotameter was also
located between the flask and the pump so accurate flow measurement was obtained.
Four of these samplers were operated along with the samplers for bacteria.
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Mixed Liquor Samples from Aeration Basin
A daily composite mixed liquor sample was collected from the second stage aeration basin during Period
4 and a daily sample was collected from the first stage aeration basin during Period 5. A model WM-5-24
Sigmamotor sampler was used for the collection of the aeration basin sample. The 24-hour composite sample
was split into two equal parts during the collection process. The first part consisted of approximately 150 ml
collected every fifteen minutes and placed in a 20-liter carboy which had been sterilized prior to use. The
second part consisted of approximately 150 ml collected every fifteen minutes and placed in a 20-liter carboy
which had been washed and rinsed with nitric acid prior to use.
The first part was analyzed for microorganisms while the second part was analyzed for chemicals (trace
metals and TOC). The carboys and samples were maintained at 40 °F in a refrigerator located below the
sampler. After every 15-minute sampling period, the automated sampler purged the collecting tubes between
the sampling point and the carboy. Every morning the collected samples were removed and an empty steri-
lized carboy and an empty acid-washed carboy were placed in the refrigerator for the next daily composite
sample. The collection tubes for the microorganisms were replaced on a daily basis and autoclaved prior to
use. The collection tubes for the chemicals were similarly replaced and acid rinsed prior to use.
During the Period 5 sampling, an additional sample was taken from the first stage aeration basin. This
sample was a one-hour composite sample that was started fifteen minutes before each aerosol run and ter-
minated fifteen minutes after each aerosol run. This sample was analyzed for microorganisms only and served
as an indicator of what the characteristics of the first stage aeration basin were during each aerosol run.
All samples collected were kept at 40 °F during shipment and analysis was begun within 24 hours after
collection.
Soil Samples
Three separate surface soil samples were collected for trace metal, bacterial, and viral analyses. A total of
50 samples for each of these three types of analyses were collected during each sampling period. The samples
for trace metals were collected using a plastic digging device to collect several grams of surface soil. This sam-
ple was placed into a polyethylene container which had been prewashed to remove trace metals. The con-
tainer was sealed and kept frozen until analyzed.
The soil samples for bacterial and viral analyses in Periods 1 and 2 were taken in the following manner.
Using a sterile digging instrument, a surface soil sample of several grams was taken and placed in a vial con-
taining thioglycolate solution and shaken. The material was then shipped on wet ice on a daily basis to San
Antonio. The soil sample for virus determination was collected in the same manner as the samples for bacteria
except that the sample was frozen immediately after collection and kept frozen until analyzed. No solution
was added to the soil sample for viruses.
During Periods 4 and 5, larger soil samples were collected for bacteria and virus analyses. The samples
were collected in the same manner as in Periods 1 and 2 with the exception that no thioglycolate solution was
used to preserve the bacterial samples.
The soil samples for trace metal, bacterial, and viral analyses were collected on the same day, sometime
during each of the four sample periods. The samples were not collected during periods of precipitation. The
locations for collection of soil samples were designed to describe the windrose pattern, but they included the
preselected air sampling sites described earlier. The remainder of samples were taken along radial grids to fill
in the gaps between air sampling sites to better define the windrose pattern.
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Water Samples
Water samples were collected from ponds, lakes, and streams located within a five-kilometer radius of
the water reclamation plant. A total of 10 water samples were collected during each sample period and
analyzed for trace metals, bacteria and virus. For Salt Creek, water samples were taken upstream and
downstream from the water reclamation plant. Water samples were taken on a single day during each of the
four sample periods.
Approximately 250-ml samples of water were collected for trace metal analysis in polyethylene containers
which had been specially washed for this purpose. These samples were frozen and kept frozen until analyzed.
A separate sample was collected for bacterial analysis using asceptic techniques. The bacterial samples were
collected by a grab technique in which one liter of water was collected. These samples were shipped on the
same day they were collected.
Water samples for the virology study were collected in a one-liter container. The one-liter sample was col-
lected in a wide-mouth screw-cap plastic bottle and frozen until analyzed in the laboratory.
Meteorological Sampling
At the plant site a meteorological station was set up and operated during the environmental sampling
periods. During the air sampling runs information was recorded for wind speed and direction, temperature,
humidity, and solar radiation. Values from these instruments were used in aerosol sampling runs and to
describe the meteorological conditions during the remaining time in the sample periods.
Supplemental climatology data was obtained from the weather bureau maintained at O'Hare Airport
located approximately eight miles east-southeast of the Egan Plant. Prior to starting a sampling run, whether
for aerosol or paniculate collection, the O'Hare weather people were called. A prediction for wind direction
and speed was obtained for the subsequent few hours to determine if conditions were applicable for aerosol
sampling. The high-volume paniculate samplers were set up based on the wind prediction for the subsequent
24-hour period. This information about the projected wind conditions and possible precipitation patterns was
used in conjunction with the conditions at our weather station maintained at the Egan plant. These two
sources provided the criteria for setting up samplers and/or moving them if necessary.
The equipment used to obtain and record the meteorological conditions in the study area was set up at
the Egan treatment plant. Measurements for wind speed and direction, solar radiation, temperature, and
humidity were recorded on strip chart recorders that were an integral component of each piece of equipment.
Wind speed and direction were monitored using a Model 6405 Field Recording Wind Set purchased from
R. M. Young Company, Traverse City, Michigan. The windvane and cup anemometer were mounted on top
of a five-meter pole. The range of the windvane was 360° mechanical, 342° electrical (5% open), with con-
tinuous rotation. The anemometer was operated in the 0 to 50 mph range with a threshold of less than 1.5
mph. The recorder-translator was powered by a 12-volt, 60 ampere-hour battery.
Solar radiation was monitored using a Model B-5-3850A Recording Pyrheliograph available from
TechEcology, Sunnyvale, California. This instrument with a sensitivity of 0.1 gram-calorie per square cen-
timeter was located adjacent to the weather tower.
The temperature-humidity instrument was placed in a shelter adjacent to the weather tower. The
Hydrothermograph Model B-5-594 available from TechEcology has a temperature range from —20° to
110°F. with an accuracy of ± 1 °F. The humidity element has a range between 20% and 95% RH with a ± 3%
accuracy.
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CLINICAL SPECIMEN SAMPLING
Organizational Overview
The design of the human sampling phase of this project required that those individuals selected as sub-
jects would remain with the project until all human tissue sampling was completed. During the two-year
sampling period, human tissue samples were collected before and after the treatment plant went into opera-
tion. Each subject then was his own control and the necessity of having all of the subjects remain in the pro-
gram was removed.
Because the study site near Chicago was far removed from the Institute laboratories in San Antonio, it
was necessary that an on-site coordinator be selected. The responsibilities of the on-site coordinator involving
the human tissue sampling were many. One key element was the availability of a person in the study area who
could maintain contact with the human subjects during actual sampling periods and in the interim between
sampling periods. Because the coordinator was a resident of the area, assistance was given to the sampling
team on items such as the availability of a biological sampling center for the subjects, where to find medical
personnel to help with the sampling, and other details that the survey team requested.
An important consideration for continued human subject participation throughout the project was the
necessity of having competent, reliable, and friendly medical laboratory technologists. Based on previous ex-
perience with human subjects, the way collection of blood was performed created a lasting impression of the
whole project and more importantly whether that subject would return to give future blood samples. Compe-
tent medical technologists to collect blood for this project were located at the Alexian Brothers Hospital.
With the help of Dr. James Clark, who was director of the clinical laboratories at Alexian Brothers Hospital,
certain medical technologists were identified that met our criteria of being competent, reliable, and friendly
technologists. These persons were approached and asked to help with the epidemiological survey.
Secondary to finding competent blood drawers was the problem of locating capable people to collect hair
samples from subjects. Similar to blood collection, the proper collection of hair samples was vital to con-
tinued subject participation. Subjects would not be expected to return if hair samples were taken in such a
way as to drastically alter the appearance of their hair. Yet the first concern was to collect the proper hair
sample in the required amounts. Barbers and hairdressers were found in the study area who could collect the
proper sample while maintaining the subject's appearance.
The nature of the project and its goals needed to be disseminated through the study area so the communi-
ty would be aware of this epidemiological study and its impact upon them. Local papers serving the area were
provided press releases that described the project, its goal, the study area, and reminders to the study par-
ticipants of upcoming sample collection periods.
Presampling Preparation
After the selection of the human subjects, letters were sent to each individual who completed a question-
naire (Appendix C). Letters sent to those people who were not selected thanked them for their willingness to
participate. They were told that their questionnaire had been set aside for the present but that they would be
contacted if any vacancy occurred.
The selected subjects were notified and given a date and place where the survey team would meet with
them. In this letter, mention was also made of the on-site coordinator and how she could be reached if a con-
flict or other problems occurred prior to meeting with the survey team.
Each subject received a collection kit that had containers for the samples to be collected at home. Prior to
meeting with the subjects the sample collection kits were assembled. The kit included a brief summary of the
39
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nature of the study, a set of instructions on the proper collection of urine, feces, and sputum samples, and a
portable camping toilet to facilitate the collection of the feces sample. Containers were provided for the
samples and each had its own unique computer-printed label that had the subject's name, type of specimen,
and an identification code for sample processing.
At the first meeting with the subjects, it was convenient to meet with them at the processing center. The
sample collection kits were handed out and instructions given on the proper collection of the samples. Ap-
pointment times were also set up for them to return to the collection center with their samples and to have
blood, hair, and throat swabs collected. During, the subsequent sampling periods, after the subjects had
familiarized themselves with the sampling routine, arrangements were made to contact them by phone. The
phone call was made by the on-site coordinator to arrange a time that the collection kits could be delivered to
their homes. Appointment times were also set up with the subjects as to when they should come to the pro-
cessing center with their samples. This procedure of delivering the collection kits to the homes and setting up
appointment times was initiated as a convenience to the subjects so they would have to come to the collection
center only one time.
Sampling Protocol and Sample Collection
The proper and complete sample collection by each participant necessitated that a sampling scheme be set
up to obtain the best result. Institute personnel and the on-site coordinator formed an integral team to ac-
complish this task. The on-site coordinator, together with help from the person designated as the biological
sampling manager, set up an informal procedure so each subject received as much attention as possible. With
this effort, each subject was made to feel that his cooperation was vital to the success of the program. The
result was that each subject did the best he could in following instructions regarding sample collection.
The first meeting with the subjects at the collection center was to explain the objectives of the project as
well as to verse each subject as to the importance of proper sample collection. During this meeting, the sub-
jects were given ample opportunity to ask questions regarding any portion of the study. Once the subjects
were fully informed, they were requested to complete an informed consent form as required by the National
Institutes of Health regulations concerning human subjects. A copy of this form is included in Appendix D.
Each subject was also asked to include the name of their family physician, if they so desired. Their doctor
would then be notified as to the presence of pathogenic organisms in their individual samples.
After the subject had completed and signed the informed consent, he was given his own sample collection
kit. The subject then set up an appointment time to return with the samples and to have the remaining samples
collected by qualified technicians.
Each subject was required to collect overnight urine, fecal, and sputum samples as well as to fill out a
current health status questionnaire. An instruction sheet was included with each collection kit in case the ver-
bal instructions given when the kits were handed out were forgotten. A copy of this instruction sheet is includ-
ed in Appendix E.
The night before the subject was to report to the collection center, he was instructed to begin collecting
his urine in the designated container. The urine samples were collected in specially cleaned wide-mouth
polyethylene containers (1/2 gallon size). The subject was cautioned against putting anything other than his
own urine into the container. The urine samples were collected from approximately 5 p.m. until 8 a.m. the
following morning, the day for blood testing. Each subject was instructed that once he began collecting his
urine he was to continue collecting all his urine until the following morning. It was also emphasized that the
first specimen upon arising in the morning must be included in the total sample. After this specimen was col-
lected the subject was told to discontinue urine collection.
An overnight fecal sample was collected using a portable camping toilet to aid in collection. The portable
toilet was provided as a convenience if the subjects desired to use it. They were instructed to begin collection
40
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of the fecal specimen the evening before their scheduled blood test. When collecting the specimen the subjects
were cautioned against mixing any urine or toilet paper with the feces in the polyethylene collection container.
If the subjects had a place to keep the feces and urine specimens cool they were urged to do so.
The collection of the proper sputum specimen was the most difficult sample to obtain. The subjects were
told that saliva was not an acceptable specimen. A sterile polyethylene collection cup was provided for the
sample and the subjects were told to open the container only when collecting their sputum. The subjects were
told that a valid sputum sample must come from inside their lungs and probably could be best collected the
first thing in the morning. Coughing deeply several times should produce the required sample.
Each subject was also given a Current Health Status Questionnaire to fill out for each sampling period.
By filling out the form at home, the subject had a more relaxed surrounding rather than at the collection
center with all its activity. The subjects were also provided extra copies of the questionnaire for them to keep a
daily log of symptoms should any occur prior to the next sampling period.
Blood was collected from each subject by a qualified medical laboratory technician under the supervision
of a physician. The blood was collected using sterile techniques into two 10-ml vacutainer tubes manufactured
by Becton Dickinson Co., Rutherford, New Jersey. The minimal lead type (L3200XF313) containing 143
units USP sodium heparin was used for metal and hematocrit determinations. A second vacutainer that was
sterilized by radiation and had no anticoagulant (SR 3200) was used to collect blood for antibody liters.
While the blood collection was maintained in as trauma-free situation as possible, it was mentioned to
each technologist before collection began that some of the subjects had not previously had a venous blood
sample taken. With this in mind the technologists were instructed to use their best "bedside manner" to
reassure each subject and to minimize apprehension on the part of the subject. This improved "bedside man-
ner" was most useful in collecting blood from the children on the project.
Once the blood had been drawn and properly labeled, the blood in the heparinized tube was thoroughly
mixed. The sterile tube was not mixed but allowed to clot. The samples were then taken to the processing
room and remained there from 30 minutes up to 2 hours before processing.
A barber and hairdresser collected the hair samples from each subject. Hair was collected from the nape
of the neck and hair of 25-mm length from the scalp. The hair collectors had a sample of hair weighing one
gram to match with the amount of hair collected from the subject. Each barber and hairdresser was instructed
as to the proper amount and location for the hair sample. They were informed of the importance of collecting
the minimum amount without drastically altering the appearance of the subject's hair style. The subjects were
similarly instructed to refrain, if at all possible, from having a haircut for at least one month preceding the
survey period. The hair samples collected were placed in scalable plastic bags with the proper label correspon-
ding to the subject and the location of the sample.
The collection of throat swabs was performed by a medical technologist. Cotton-tipped applicators in
sterile 16 x 150-mm screw-cap tubes (Falcon 2021) were used to collect the samples. Where necessary, tongue
depressors were used so the proper sample from the back of the throat could be taken. Each subject had two
cultures taken — one for bacterial determinations and the other for viral determinations. Each tube was ap-
propriately labeled and placed in the test tube rack designated for viral or bacterial assay.
The collection center was set up to provide the subjects with a central location to drop off their samples
and have the remaining samples taken. In each survey period approximately 225 subjects were processed
through the center in a two-day period. This number of subjects passing through the collection center meant
that an efficient operation was necessary to give each subject as much personal attention as possible while
keeping the time spent at the center to a minimum.
The collection center was divided into six areas that each subject visited. Upon arriving at the center with
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their samples, the subjects first stop was at the "sample turn-in station." Here the survey workers checked
each sample to make sure that it was properly labeled, of sufficient quantity, and that all samples were col-
lected. An index card for each subject was kept on file to assist in the sample check-in. The subject was then
given the card which he presented at each station until he had passed through all stations of the collection
center. The index card had the subject's name, address, and identification number, and boxes for the survey
worker to initial after each sample was taken.
Following sample check-in, the current health status form of each subject was checked. The survey
worker scanned each form for completeness, accuracy, and legibility. When the questionnaire was complete,
the survey worker initialed the subject's index card and showed him to the waiting area where chairs were set
up. This area was adjacent to sample check-in but was removed from the remaining collection areas. This
kept congestion in the sample collection rooms to a minimum.
The orderly flow of subjects through each collection station was the job of the survey worker designated
as the traffic director. His job was to make sure that no collection station had a back-up of waiting subjects
while another station had no subjects to sample. Most of the time the subjects met their scheduled appoint-
ment times to the minute and this reduced any waiting time. This meant that after the sample check-in and
questionnaire stations, the traffic director took them immediately to one of the remaining collection stations.
The collection areas for hair, throat swabs, and blood samples were each in separate rooms off a com-
mon hallway. Chairs were provided in each-room for the sampling. Extra chairs were provided in the hallway
for any subjects to wait.
The hair collection room was the first area usually visited by the subjects. Four barbers or hairdressers
collected the samples from the subjects. If all the barbers were busy and some people were waiting, then the
subject either could wait at that area or go to another collection station. The throat swab station was the least
busy and the subjects did not have any waiting time. One medical technologist could easily handle the collec-
tion of the throat swabs.
The blood collection station located adjacent to the throat swab station had four medical technologists
collecting blood samples. If any problems arose in collecting blood (especially from children) then one of the
four technologists assisted in the collection.
At each collection station the survey worker initialed the subject's index card in the proper box indicating
that sample had been collected. The survey worker also mentioned to the subject what samples needed to be
taken. When the subject had visited all of the sample collections he was directed to the last station.
At the check-out station a survey worker reviewed the subject's index card to be certain that all samples
had been collected. If a sample such as feces or urine had to be dropped off later, arrangements were made as
to the time and place. The second aspect of this collection station was payment of the subjects. Each subject
was paid at the end of each collection period in the project. The survey worker assigned to this station was the
on-site coordinator. She, therefore, could answer most of the subject's questions and if not, a member of
SwRI was available to assist her. Photographs of the first sample period, both environmental and human
samplings, were displayed on a poster board at the payment station. This provided the subjects with a view of
the overall survey and how their participation fit in the whole survey.
At the last sampling period, the subjects were given a certificate of appreciation. This certificate was an
expression of thanks on behalf of the survey team for the cooperation and participation of each participant.
The certificate was printed on parchment paper and could be framed and hung in a suitable place by the sub-
ject. A copy of this certificate is attached in Appendix F.
The containers used to collect the samples required various cleaning methods and handling techniques to
prevent contamination. The polyethylene containers for feces and urine collection and the blood vials used in
42
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transporting the sample were cleaned as follows. Each container was washed with a laboratory detergent
(Alconox) in tap water. Then it was rinsed with deionized water and placed in an acid vat containing HNO3
(1:1) and allowed to soak for 6 to 18 hours. After acid soaking, the items were rinsed thoroughly with deioniz-
ed water and placed in a 60°C drying oven. The dried items were then capped and stored until shipment to the
survey site.
The containers used for collection of throat swabs and sputum specimens required no cleaning as they
were bought from the manufacturer in sterile packages. The survey workers made certain that each container
or tube was tightly closed before the sealed bags they were in were opened. This precaution was also used with
the sterile tubes used in transporting the serum used for antibody liters.
The hair samples were collected and transported in Zip-Lok bags. The bags were checked by lot number
to be certain that no metal contamination was present, and were then used without further cleaning.
Sample Processing
Management and Special Processing-
The nature of the sample processing required that some processing be performed at the collection center
and some away from the collection center.
Teams of medical technologists were assembled at the collection center and the processing center to
handle sample processing. Before any team member began working with the samples, the biological process-
ing manager briefed all the sample processors. Emphasis was placed on the proper handling of the specimen
to prevent contamination. Other points covered during the briefing included the proper container for the sam-
ple, the labeling system, and the storage of the sample after processing.
Prior to processing, the samples had labels with the subject's name, his own unique identification
number and what the sample was, i.e., urine, feces, or blood. Once the sample processing began, the labels
used had only the subject's identification number plus a code designating the type of further processing. Sam-
ple codes were used to distinguish blood for trace metal or antibody analyses, feces for parasite,
microbiological, or trace metal analysis and urine for trace metals. The processing labels thus provided con-
fidentiality and a means for computer processing.
At the collection center one room was set up to process the blood samples. This room, while near the col-
lection activity, was kept closed to the subjects. The blood samples were brought to this room on a routine
basis.
The urine and feces specimens turned in by the subjects at the first station were kept in closed ice chests.
These samples were taken to the Hanover Park Treatment Plant laboratory where processing took place. One
person was assigned to transport the samples from the collection center to the remote processing center.
Blood was collected from the subjects for metal analysis and for microbiological determinations. The
samples were taken to the processing room at the collection center on a routine basis. The blood samples for
the antibody liters were allowed to clot and then were cenlrifuged. The serum was laken off using a slerile
disposable pipel (Falcon 7536), placed in a 12 x 75-mm slerile polyelhylene lube (Falcon 2005) and secured
wilh a cap. The processing label corresponding lo Ihe sample label was affixed lo Ihe lube which was ihen
placed on dry ice.
The blood sample collecled in Ihe tube wilh anticoagulant was used for hematocril and melal delermina-
lions. Each blood sample was placed on an aliquot mixer and thoroughly mixed before blood was laken for
Ihe hematocrit delerminalion. A Clay-Adams hemalocrit centrifuge was used lo spin Ihe capillary lubes. The
hemaiocril value was then recorded on Ihe dala sheel.
43
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The throat swabs collected from each subject were treated with two types of growth media. The
technologist taking the throat swab added Brain-Heart Infusion broth (BHI) to the tube designated for viral
analysis. Thioglycollate broth was added to the tube designated for bacterial analysis. The appropriate'broth
was added to each tube so the cotton applicator was thoroughly moistened and some excess was in the tube.
The media was added to the tube as soon after collection as possible to prevent the sample from drying out.
Blank throat swab tubes were also treated with the growth media and served as quality control samples to
identify any contamination.
Urine samples collected by the subjects were processed at the remote laboratory facilities. Each urine
specimen was weighed and the amount recorded on the data sheet. An aliquot (175 ml) of the thoroughly mix-
ed urine was placed in a clean polypropylene bottle. One-percent (1.75 ml) acetic acid was added and the bot-
tle labeled and placed on dry ice.
The feces specimen required a number of steps in processing. The total sample was first weighed and the
weight recorded. The specimen container was then opened and the aliquots taken. Using sterile sticks three
areas of the stool specimen were sampled. Approximately one gram of sample was placed into each of three 4-
dram vials. The first vial had thioglycollate broth as a bacterial nutrient, the second had BHI as a viral
nutrient, and the third had merthiolate for parasite preservation. Prior to adding the feces to the merthiolate,
0.5 cc of a freshly prepared iodine stock was added to the vial. In each case, the feces added to vials was
thoroughly macerated with the sterile sticks. After the vials were capped, they were vigorously shaken to pro-
mote a homogeneous solution. Each vial was then labeled and put into separate wooden racks designated for
parasitic, viral, or bacterial analysis. The bacterial vials were secured in the racks and made ready for ship-
ment. The viral samples were frozen and the parasite samples maintained at ambient temperature. The re-
mainder of the feces specimen was frozen without any added preservative and analyzed for trace metals.
These specimens were maintained under those conditions until shipment was made to the San Antonio
laboratories.
One feces sample was also aliquoted into vials for a quality control check of the viral analysis. These vials
were labeled similar to those used for the other subjects' samples and could not be distinguished from the ac-
tual sample labels. The only exception was that the samples were spiked with poliovirus and had unused labels
prepared for subjects who did not participate during the sampling period.
The hair samples collected by the barbers and hairdressers did not require any further processing at the
study site with the exception of packaging all of the bags with the hair samples into one box.
The sputum samples to be analyzed for mycobacteria were placed into insulated mailers as the samples
were turned in at the collection center. The samples were sent in these mailers to the San Antonio laboratories.
Preservation and Shipment-
All the time of sample processing the shipping and subsequent storage conditions were established for
each sample. The following table lists the human samples collected and the means of preservation other than
chemical.
Sample Type Analysis Shipment Conditions
Blood Trace metals — 78 °C
Antibody liter — 78 °C
Urine Trace metals —78 °C
Feces Bacteria 4°C
Virus — 78 °C
Parasites Ambient
Throat swabs Bacteria 4°C
Virus — 78 °C
Sputum Mycobacteria 4°C
Hair Trace metals Ambient
44
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Following the collection and processing of samples, specific types of samples were either shipped im-
mediately or held for one shipment of that type of sample. The samples that required immediate shipment
were the throat swabs and feces for bacterial analysis and the sputum specimens. These samples were sent by
air at the end of the sampling day. A Kool-Pac was placed in each insulated mailer to keep the samples cool
until analysis.
Prior arrangements were made so the samples shipped from Chicago in the early morning hours (about 1
a.m.) would be picked up at the San Antonio airport upon arrival.
The frozen samples awaiting shipment were kept either in a freezer or in ice chests packed with dry ice.
Approximately one week before the shipment of all the frozen samples, the airline's freight manager at
O'Hare was contacted to reserve a specific flight. The samples were shipped in dry ice and could not occupy
the same freight area on the plane with live animals.
The remaining samples such as the feces for parasite analysis and the hair samples were also in the ship-
ment with the frozen samples but separate from them. The remaining supplies or equipment was sent back to
the San Antonio labs via truck along with the equipment used for the environmental survey.
Procedural Changes-
Prior to the last human sampling period, the procedure for collection of feces for parasite investigation
was modified. During the previous sampling periods, feces specimens were collected at home by the subjects
and brought to the collection center. The samples were then aliquoted to the various vials filled with media at
the processing center as previously described. The change of the procedure involved a set of 50 subjects
selected at random, living throughout the study area. They were given a cup to collect a feces specimen. A vial
with the merthiolate was poured into a labeled vial with the iodine just prior to the subject sampling the feces
specimen with the sterile sticks provided. A small amount of the feces was placed into the vial with the mix-
ture. The contents of the vial were stirred until a homogeneous sample was obtained. The vial was capped and
brought to the collection center at the time of their scheduled appointment. No further processing of the sam-
ple took place at the processing center other than to include these specimens with the other samples designated
for parasitic analysis.
ANALYTICAL METHODS
Organizational Overview
Organizations responsible for various portions of this study are listed in Table 2 in Section 1 of this
report. Each organization was selected because of its outstanding reputation in that area of endeavor. The
responsibility for analysis of environmental aerosol and sewage samples was changed to UTSA during the last
sampling periods due to their heavy involvement in programs of a similar nature for both EPA and the U.S.
Army Research and Development Command.
Aqualabs was selected not only because of its good reputation, but because of its location close to the
study area. In order to obtain reliable data it is necessary that certain analyses such as standard plate count,
fecal coliform and total coliform be begun as soon as possible after collection. Aqualabs was able to meet our
schedule which called for sample delivery after normal working hours.
Microbiological Analysis of Environmental Samples
Bacteriology-
Mixed Liquor Samples from Aeration Basin-The aeration basin samples were analyzed by procedures
detailed in Standard Methods, either 13th edition(22^ or 14th edition(23). Total and fecal coliform were deter-
mined using the membrane filter method^, and standard plate count was determined according to Standard
Methods*23^ using an automatic colony counter.
45
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Isolation of microorganisms was conducted by inoculating samples onto S-S Agar, Endo Agar, Mc-
Conkey's Agar, and Blood Agar. Lowenstein-Jensen Medium and Middlebrook 7H-10 Medium were used for
isolation for Mycobacterium species. Identification of microorganisms was by procedures and techniques as
given in Manual of Clinical Microbiology(24), and Bacterial and Mycotic Infections of Man(25). Cultural mor-
phology, biochemical reactions, microscopic appearance, and serologic testing were employed as required.
Composite Samples-Analyses for fecal streptococci were performed in accordance with standard pro-
cedures and techniques that have been established for identification of these organisms in Standard
Methods'2^. The membrane filter technique was employed as this procedure yields more reproducible results
than does MPN.
Fluorescent pseudomonads (i.e., Pseudomonas aeruginosa, P. putida, and P. fluoresens) were determin-
ed by the following procedure. A direct and quantitative approach was found to be more useful for their
enumeration than the MPN analysis, since fluorescent pseudomands were present in large numbers in the
large-volume composite and aerosol samples. Samples were diluted and plated in triplicate onto previously
solidified plates of King's medium B, commercially available as Pseudomonas Agar F (Difco), and onto
plates of Centrimide Agar (Difco). The former medium is optimal for production of fluorescent pigments
(pyroverdin) by pseudomonads and the latter is a selective medium for Pseudomonas. Following incubation
at 37 °C for 48 hours, the number of fluorescent pseudomonads was determined by noting the number of
green fluorescing colonies observed with long-wave ultraviolet light in the dark. When the Pseudomonas Agar
F plates were excessively crowded with other types of organisms, all colonies from appropriate dilutions were
picked from the selective pseudomonad medium (Centrimide Agar) to sectors of Pseudomonas Agar F plates.
Following incubation, the number of fluorescent colonies were confirmed by demonstrating that they were
oxidase positive.
Enrichment for Salmonellae (including Arizona) was carried out following concentration of samples by
filtration of measured volumes of sample (the volume filtrable depending on the level of organic and par-
ticulate matter) on diatomaceous earth plug and an unconcentrated 10-ml sample were enriched in separate
bottles of selenite and tetra-thionate broths. After enrichment, aliquots were streaked for isolated colonies
onto a moderately selective medium (XLD or Hektoen enteric agar) and a highly selective medium (SS). Iden-
tification of suspect colonies was accomplished by a biochemical screen (oxidase, Triple Sugar Iron, Motility,
Indole, Ornithines, Lysine, Urea). Additional biochemical tests were performed on all colonies with a ten-
tative identification of Salmonellae.
A qualitative procedure for the concentration and isolation of Shigella was carried out as described in
Standard Methods^23'. Filtration through diatomaceous earth was used to concentrate a large proportion of
all microorganisms present in a 1- to 2-liter sample. The resulting plug of diatomaceous earth was divided into
equal portions for addition to bottles of GN broth (BBL). One-half of the same plug also was used for enrich-
ment of Salmonella as described above. A 10-ml portion of the unconcentrated sample was added to a
separate bottle of enrichment medium. After 24 hours enrichment at 37 °C, aliquots from the bottles were
plated for isolated colonies onto xylose lysine desoxycholate (XLD) agar. Although several enrichment-
plating media combinations are available for isolation of Shigella, the GN-XLD combination repeatedly has
been observed to be superior for recovery of the organism.
Proteus was determined by directly plating serial dilutions of the sample onto moderately selective and
highly selective enteric media. All colonies were oxidase tested; on those which were negative, a preliminary
biochemical screen was performed. All colonies tentatively identified as Proteus were subjected to further
biochemical tests for confirmed identification.
Large Volume Microbial Characterization Samples-
Large volume samples were collected both at the beginning of the study and at the end. They were intend-
ed to provide information that would assist in examining the possible relationship which might exist between
routine indicator organisms and selected pathogens.
46
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A number of investigators have reported that staphylococci have a greater resistance to residual chlorine
than do the coliforms and fecal streptococci. Staphylococci as an indicator of pollution in swimming pools
may be primarily a measure of pollution arising from the bathers' eyes, ears, nose, and throat. However,
staphylococci also are a component of the normal gut flora of man and many provide a useful measure of
fecal pollution, particularly in treated waters. In early parts of the study, a quantitative membrane filter (MF)
procedure described in Standard Methods^23) was employed. The sample size varied in order to yield MF
counts within a desired 400-100 colony range on M-Staphylococcus broth-soaked absorbent pads. However,
enumeration of yellow pigmented colonies and their subsequent confirmation as S. aureus was complicated
by the heavy mixed growth on the filters. This procedure was abandoned and samples (0.1 ml) were plated in
triplicate directly to previously solidified plates of Mannitol Salt agar. This change resulted in a marked
reduction of "background" growth, but reduced the volume of sample that could be analyzed. Typical col-
onies showing a yellow zone of mannitol fermentation were fished for confirmation of identity by staining
techniques and coagulase production.
Tubercle bacilli survive for long periods in feces, sewage, and water and have a greater resistance to
chlorination than the coliform indicator group. Mycobacteria were assayed quantitatively by a procedure
which almost totally suppresses sewage saprophytes while permitting recovery of most Mycobacteria. A large
sample was treated for 20 minutes with 500 ppm of Benzalkonium chloride (Zephiran), diluted and plated to
the surface of previously prepared plates of Middlebrook 7H10 Agar. Plates were incubated with five-percent
CC>2 and examined over a period of one month for the appearance of colonies of Mycobacteria. Colonies that
appeared were identified by examination of stained (Ziehl-Neelsen) smears for acid-fast bacilli and by other
procedures as outlined in Figure 7.
Members of the genus Leptospira have been implicated in a variety of human diseases. Entry into the
host generally occurs through abrasions of the skin or through mucosal surfaces following contact with water
contaminated with urine of infected humans or wild and domesticated animals. Pathogenic leptospires may
survive for months in neutral or slightly alkaline waters. "Saprophytic" or "water" leptospires (L. biflexa)
are found predominately in fresh surface waters and are rarely associated with infection. The assay for lep-
tospires was accomplished by their initial separation from contaminating microbes by passage of the test
samples through 0.45//. membrane filters. Leptospires will pass through the filters because of their small
diameters. The filtrate and unfiltered samples were diluted into tubes of Stuart's liquid medium (with 10- per-
cent inactivated serum) containing 200 jig/ml of 5-fluorouracil to inhibit the growth of contaminating
microorganisms. The tubes were incubated in the dark at 28 °C. Cultures were examined at 5- to 7-day inter-
vals over a total period of six weeks for growth and microscopically (dark-ground microscopy) for the
characteristic leptospiric morphology and motility.
Spores of the anaerobic Clostridium perfrigens (Cl. welchii) are very resistant to disinfectants and other
detrimental environmental effects and survive longer in water than conventional indicator organisms and
most pathogens. This organism was determined by the test recommended by the World Health Organization.
Aliquots of decimal dilutions of the test samples were inoculated in triplicate into freshly boiled litmus skim
milk medium (with 0.1 % "reduced" iron powder).
The tubes were heated at 80 °C for 15 minutes to destroy vegetative cells, incubated at 37 °C and ex-
amined over a period of five days for positive "stormy fermentation" reactions. To decrease the incidence of
false positive tubes, the tests were confirmed by examination of wet mounts and stained smears for non-
motile, encapsulated, Gram-positive rods. Spores, if present, were located centrally within the cells. Com-
putation of the MPN index was accomplished by consulting probability tables used for estimation of col-
iforms.
As emphasized in Standard Methods(23), differentiation of the genus Klebsiella from other coliform
organisms is important because of its association with human respiratory and genitourinary infections. Quan-
titation of Klebsiella in the highly mixed samples was complicated by the lack of an isolation procedure that is
47
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1. Sample Processing
Treatment with 500 ppm Benzalkonium chloride
(Zephiran) for 20 minutes
solution plating
Middlebrook 7H10 Agar Plates
CO; Weekly observation - four weeks
2. Selection of Colonies
Henry Oblique Transmitted Light,
Law Power Stereomicroscopy
Colony Characteristics (Amount and kinds of roughness,
shape, pigmentation, etc.)
3. Preliminary. Screen
Zichl-Neelsen Stain
Acid-fastness, cellular characteristics
Acid-fast Bacilli
4. Subculture
Tubes of Lowenstein Media; W~
5. Further Tests for Identification
Figure 7. Schematic diagram of mycobacteria isolation.
48
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highly differential or selective for these organisms. The capacity of Klebsiella and Enterobacter to fix nitrogen
and thus to form large colonies on a nitrogen-deficient medium has been employed for the differential isola-
tion of these organisms from feces. However, the most commonly employed ability of Klebsiella to produce
large mucoid colonies on differential enteric media containing a high concentration of carbohydrate (e.g.,
EMB agar) was utilized in the present study.
The isolation and identification scheme employed for Klebsiella is summarized in Figure 8. Klebsiella
ozaenae and the rare K. rhinoschleromatis were screened for in addition to the more common K. pneumoniae.
Most Klebsiella in nature are encapsulated and give rise to mucoid colonies, but rough forms also occur.
Finally, the closely related genera Enterobacter and Serratia, that "classically" produce less mucoid colonies
than Klebsiella, cannot be differentiated from the latter on the basis of colonial morphology alone.
The above considerations and the large number of mucoid colonies observed following plating of the
samples to the nonselective enteric media, made it apparent that quantitation of Klebsiella would have to be
carried out in an indirect manner. Individual mucoid colonies, randomly picked from composite and aerosol
samples plated throughout this study to EMB, MacConkey, and ENDO agars, were tested for identification
as Klebsiella. From this analysis, EMB was selected as the medium of choice for determination of total
mucoid colonies.
Members of the other genera of Enterobacteriaceae (i.e., Enterobacter, Seratia, Edwardsiella,
Escherichia, Citrobacter, Providencia and Yersinia) were determined by picking colonies from samples dilu-
tion plated to moderately selective (XLD and Hektoen) and highly selective (Bismuth Sulfite and SS) enteric
plating media. Two-to-three representatives of every colony type observed under low-power microscopy with
oblique transmitted lighting were subcultured for identification as illustrated in Figure 8.
All colony types picked for identification following enrichment for Salmonella and Shigella as described
before also were characterized to the species level for inclusion in this aspect of the study. Colonies from the
nonselective enteric plating media (EMB, ENDO and MacConkey) were not selected for this aspect of the
study. Since the coliforms (particularly E. coli) constituted the major portion of Enterobacteriaceae on the
nonselective media, their light-to-heavy suppression on the selective media was desirable to increase the
chance of picking other types.
Yersenia enterocolitica was determined by selective enrichment of membrane filter-concentrated samples
or diluted samples refrigerated in isotonic saline with 25 /tg/ml of potassium tellurite and plated to SS agar.
After incubation for two days at 25 °C, colonies were identified as described previously.
Assay for total and fecal coliform was conducted as prescribed in Standard Methods.
Aerosol samples-The procedures employed for isolation and identification of bacteria from aerosol
samples in Periods 1, 2 and 4, whether the organisms were collected directly onto Blood Agar Plates,
MacConkey's Agar Plates, S-S Agar Plates and Middlebrook 7H-10 Medium Plates or impinged and trapped
in a fluid vehicle for transport to the laboratory, were standard laboratory procedures as referred to above.
Bacterial counts were determined on the plates which had been shipped from Chicago, and suspicious col-
onies were picked for identification. Fluid samples were plated in the laboratory and isolates selected for iden-
tification by the techniques discussed previously in this Section.
The methods employed in bacterial analyses for Period 5 aerosol samples were identical to those
employed in the analysis of effluent samples with the exception that the sample volumes were larger for the
aerosol samples. Further, the direct spread plate technique was used for fecal streptococci.
Soil Sa/np/es-Bacteriological examination of soil samples, after the soil was suspended in water, was
performed by procedures similar to those outlined previously in this Section.
49
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- Direct Plating -
(For KlebsieTla) (Other Enterics)
(1) Sample Processing
- Enrichment -
(For Salmonella, Shigella)
(For Yersinia)
Non- Mod.
Selective Selective
Highly
Selective
EMB, Mac Conkey XLD,
Endo HEK
BS, SS GN, Tetrathionate
Selenite
4-
XLD, SS
(2) SELECTION OF COLONIES
Henry Oblique Transmitted Light
Heart Infusion Agar (HIA)
1
(3) PRELIMINARY SCREEN
Oxidase, TSI, MIO (Motility-Indole-Ornithine), Lysine, Urea
J.
(4) FURTHER TESTS
Saline + Potassium
Tellurite, 4°C
*
SS (RT)
Oxidase+
Oxidase-
Fermentative (F)
I
Citrate, Esculin,
Arabinose, Inositol
Non-Fermentative (NF)
Fermentative (F)
Non-Fermentative (NF)
Oxidation-Fermentation,
Citrate, Esculin,
Pseudomonas F, Nitrite
Additional Tests*
I
Oxidation-Fermentation,
Citrate, Esculin,
Pseudomonas F, Nitrite
Arabinose, adonitol, arginine dihydrolase, citrate, esculin, JTR (Jordans tartrate), KCN, malonate, MR, MP,
mannitol, dulcitol, rhamnose, salacin, sorbitol, motility (wet-mount and/or semisolid agar), indole from
tryptone broth, enterotube, salmonella antisera, shigella antisera.
Figure 8. Schematic diagram of enterics isolation.
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Water Samples~T\\e procedures employed for isolation and identification of bacteria from water
samples were similar to the procedures outlined previously in this Section.
Virology-
Mixed Liquor Samples from Aeration Basin-The aeration basin samples from Period 4, received in one-
half gallon bottles, were concentrated by the method described as the "sandwich" technique. Briefly, this
consists of drawing the fluid sample through a sandwich of a mixture of Celite (Diotomaceous Silica) and
Talc between two Tween-80- treated and distilled water-rinsed 42-mm prefilters. The concentrated virus is
then eluted from the Celite-Talc sandwich by forcing through it 4-ml of 2 percent casein in 0.5 M Tris, pH 8.3.
Isolation of viruses from the concentrates was performed by procedures given in Diagnostic Procedures, Viral
and Rickettsial Infections(26), Viral and Rickettsial Infections of Man(27), and procedures of the NIH/WHO
Regional Reference Center for Simian Viruses established at this institution. These procedures provide for
demonstration of cytopathology in primary kidney cell monolayers and continuous cell cultures (WI-38) and
detection of plaques by an agar overlay on cell monolayers in one-ounce prescription bottles.
Mixed liquor samples from Period 5 were separated so that the solid and liquid fractions could be
assayed independently for both coliphage and enteroviruses. The samples were separated by low-speed cen-
trifugation (5,000 x g) for five minutes. The liquid fraction was decanted and concentrated (see below) or us-
ed for direct plating. The pellet was resuspended in tryptose phosphate broth and homogenized in a tissue
grinder prior to further centrifugation and assay.
All liquid samples were prepared for virus assay by the bentonite concentration method, which follows.
Two-hundred mg of bentonite clay and 20 ml of 0.01m CaCh were added to two liters of sample. The pH was
adjusted to 6.0 and the sample mixed with magnetic stir bars for 30 minutes. The suspended solids were col-
lected and pelleted by light centrifugation. The supernatant was discarded, the solids resuspended in tryptose
phosphate broth, eluted for five minutes, and centrifuged again. The supernatant (approximately 10 ml) was
decanted for assay. In the case of large volume samples, four liters were concentrated.
As a daily control, a known liter of Poliovirus Type I(CHAT) was added to 0.5 liter of the liquid fraction
of the samples and concentrated. This procedure served to determine the concentration efficiency for that
day's sample as well as cell sensitivity.
Viruses were enumerated routinely on Hela cell monolayers and reported as pfu/liter original sample
(solid or liquid fraction).
Assays were carried out using 0.2-ml samples on Hela cell monolayers grown on 60-mm culture dishes in
Eagle minimal essential medium plus 200 units of penicillin G per ml, 16.6 units of streptomycin per ml, 25 ^g
of gentamicin per ml, and 0.5 fig of Fungizone per ml.
Two-thirds of the plates of each assay were incubated three days and scored for plaques. To avoid
overgrowth of slower growing enteric viruses by poliovirus plaques, the remaining one-third of each sample
was mixed with pooled antipolio serum (types 1, 2, 3) diluted so that final antiserum concentration was 1:50.
This level was shown experimentally to reduce stock virus liter by 95 percent or more. The virus concentrale-
anlipolio serum mixture was plated, incubaled for five days, and slained lo visualize plaques.
Incubalion of all Hela cell monolayers was conducled al 37 °C in a five-percenl carbon dioxide-
humidified almosphere. After incubation, the monolayers were stained with two-percent neutral red in
Hanks' balanced salt solulion for iwo to three hours afler which plaques were counled.
Coliphage samples were assayed using 18-hour E. coli K-13 as hosl organism; this was shown lo be the
most sensilive E. coli stain tesled. Samples were prepared as described above. Results were reported as
pfu/liter original sample (solid or liquid fraction).
51
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Appropriate volumes (0.2, 0.3, or 0.5 ml) of the wastewater and 0.2 ml of host cells were added to 2.8 ml
of liquified tryptose phosphate semi-solid agar (0.9 percent) and poured while warm (43 °C) on 100-mm
plastic petri dishes prepared with 20 ml of solidified tryptose phosphate agar (1.5 percent) base layer. When
firm, the plates were inverted and incubated at 37 °C for approximately eight hours prior to counting. For
each sample fraction, a minimum often plates was used.
Aerosol Samples-The aerosol samples from Period 4 which had been collected by some form of impinge-
ment in fluids were concentrated and isolation of viruses performed by procedures described previously in this
Section.
Due to the volume of aerosol collection fluid available for assay in Period 5 (25 ml), no concentration
procedure was used. The entire volume was assayed as described above except that each 1.0 ml of sample was
used to inoculate Hela monolayers on 100-mm plates. The sample was seeded on the Hela monolayers in the
presence of 2X antibiotics. After 60 minutes for attachment, the inoculum was removed by aspiration and the
infected monolayers rinsed twice with HBSS with 2X antibiotics prior to overlaying with agar. Approximately
50 percent of the plates were incubated for three days and scored; while the remaining number of plates were
incubated for five days and scored.
Any areas of cytopathic effect (CPE) having characteristics of viral infection at three days and five days
post-infection and not directly attributable to microbial contamination, were isolated using capillary pipettes.
Hanks' balanced salt solution containing IX antibiotics (Penicillin and Streptomycin, Gentamicin, and
Fungizone) was added to each tube at 0.5 ml/tube. Such isolates were held at 4°C for at least 24 hours before
infection of tube cultures.
Confluent monolayers of Hela cells grown in stationary tube cultures were infected with 0.2 ml of poten-
tial viral isolates. After an infection period of 1.5 hours during which time the inoculum was periodically
redistributed over the cells, the inoculum was removed and 3 ml of prewarmed maintenance media containing
IX antibiotics was added. Infected cultures were returned to a 37 °C CO2 incubator.
Tubes were observed for CPE under 40X magnification on a daily basis over a period of five days. Any
tubes showing destruction of the cell monolayer were harvested by three cycles of alternate freezing-thawing
using a dry ice-ethanol bath and a 45 °C water bath. Cellular debris was pelleted by low-speed centrifugation.
Supernatants were collected (pH adjusted with NaHCO3) and stored at 4°C. This procedure constituted first
passage of isolates. Tubes which showed no CPE at five days were harvested as described and passed as
"blind" isolates. Uninfected controls also were harvested and served as second passage controls.
Prior to infection of cells for second passage, all first passage supernatants were diluted approximately
1:2 with IX HBSS and centrifuged at 15,000 x g (11,000 RPM) for 15 minutes. Supernatants from this
"high-speed" centrifugation were transferred to sterile tubes and stored at 4°C.
Confluent monolayers of Hela cells were infected with first passage supernatants (0.2 ml/tube). After a
1.5-hour attachment period, the inoculum was removed and 3 ml of maintenance media containing 2X an-
tibiotics were added. Infected cultures were returned to 37 °C CO2 incubator. Cultures were observed on a
daily basis for CPE and handled as previously described. Any tubes harvested as positive from second passage
were subjected to further verification by a three-day and five-day plaque assay. All isolates positive by this
procedure were identified using the Lim-Benyesh-Melnick typing pool.
The total available volume of BHI + 0.1-percent Tween 80 (25 ml) was plated for coliphage in a manner
similar to that described above. The only procedural change was that the 25 ml sample was distributed over
twenty 100-mm plates.
So/7 Samp/es--Virologic examination of the suspended soil samples was performed by procedures similar
to those outlined for examination of aeration basin samples and also by direct examination without concen-
52
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tration other than centrifugation to precipitate the soil particles. The supernatant fluid was then inoculated
directly onto the cell lines as above.
Water Samples-The procedures employed for isolation of viruses from water samples were similar to
those outlined for examination of aeration basin samples above.
Trace Metal Analysis of Environmental Samples
General-
Instrumentation-All analyses were performed on either a Perkin-Elmer Model 503 Atomic Absorption
Spectrophotometer (AAS) or a Perkin-Elmer Model 306 AAS. The Model 306 AAS is modified (Perkin-
Elmer Modification Kit 040-0286) to reduce "stray light" from reaching the photomultiplier tube during
operation of the flameless sampling devices.
Both AAS units are equipped with a Deuterium-arc background corrector which corrects for nonspecific
absorption. The background corrector was routinely used on all analyses.
Absorption peaks were recorded on a Perkin-Elmer Model 056 Recorder with a 10-mv range.
Flameless analyses were performed with the following graphite tube furnaces: (1) a Perkin-Elmer
HGA-2100 with the Model 503 AAS, (2) a Perkin-Elmer HGA-2000 with the Model 306 AAS, and (3) an
Instrumentation Laboratory IL-455 with the Model 306 AAS. Flame analyses on both AAS units were by air-
acetylene flames using a single-slot, 10-cm Burner Head (Perkin-Elmer Model 303-0418).
Calculation of analytical curves, regression analysis, and concentrations were performed on a Hewlett-
Packard Programmable Calculator Model 9810A (H-P 9810A).
Mercury analyses were performed with a UV Monitor Model 1235 (Laboratory Data Control, River
Beach, Florida).
Reagents-All reagents used for the preparation and analysis of the samples on this contract were of
analytical grade or better.
Metal standard solutions (1000 ppm) were from Fisher Scientific (Houston, Texas) and Ventron Cor-
poration (San Leandro, California).
Organic solvents decane and methyl isobutyl ketone (MIBK) were from Eastman Kodak (Rochester, New
York).
The chelating reagent, ammonium pyrrolidinedithiocarbamate (APDC) was from Aldrich Chemical
Company (Milwaukee, Wisconsin).
Triton X-100 surfactant was from J. T. Baker (Phillipsburg, New Jersey).
Contamination Control-A major problem in trace metal analysis is contamination of glassware, reagents,
and samples with the metal(s) being analyzed. Minimizing this problem requires an extensive control program
involving glassware cleaning, protection, and quality control measures.
All glassware and polyethylene containers that come in contact with samples or reagents are cleaned by
the following procedure: Items are washed thoroughly with a laboratory detergent (Alconox, Inc., New York)
in tap water. The clean glassware is rinsed with deionized water and placed in an acid vat containing
HNO3(1:1) and allowed to soak for 6 to 18 hours. Clean polyethylene containers are also placed in the acid vat
53
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but are removed after 4 to 6 hours. After acid soaking, the items are rinsed thoroughly with deionized water
and placed in a drying oven until dry. The dry items are placed in a dust-free area and allowed to cool.
Polyethylene containers are capped and sealed in polyethylene bags until ready for use. Glassware is returned
to its proper container (see below) and stored in glassware cabinets until ready for use.
All glassware items are kept in polyethylene containers to minimize exposure to dust in the laboratory.
Each container is numbered and contains one type of glassware (i.e. watchglasses, 5-ml volumetric flask,
etc.). When all the glassware in a container has been used, it is returned to that container and carried through
the washing procedure (see above) as a unit. While the glassware is being washed the container is also washed.
Once, the glassware has completed the wash cycle and dried, it is returned to the proper container. Several
items (three to seven) of glassware are removed from the container at this time for quality control checks.
The number of each container of glassware going through the wash cycle is entered into a log book.
Other information, such as name of technician performing washing procedure, type of glassware, length of
acid-soaking, etc., is also recorded in this log book. This allows the glassware removed for quality control
checks to be identified with a particular set of glassware being used in the laboratory. The glassware removed
for quality control purposes is checked by rinsing with a known volume of 0.1 N HNO3 and comparing with
the same acid that has not been used for rinsing. Normally one metal (Cd) is used for quality control checks
but other metals may also be included if needed. The graphite furnace (AAS) is used for quality control
analysis. Glassware which shows a significant difference between the used and unused rinsing acids metal con-
tent is referred back to the container number from which it came. That container is then returned to the
washroom and the wash cycle repeated on all of its glassware.
Mixed Liquor Samples from Aeration Basin-
Digestion of the mixed liquor samples for Cd, Cu, Pb, and Zn analyses were based upon the digestion
procedures recommended by EPA for total metal analysis of wastewater samples.
A 50-ml aliquot of the sample was pipetted into a 250-ml Vycor beaker and 20 ml of redistilled HNO3
added. The beaker was placed on a hot plate (80°C) and the sample evaporated to near dryness without letting
it boil. Another 3 ml of HNO3 was added to the sample residue and the beaker was again heated with a watch-
glass in place to create a gentle reflux condition within the beaker. From time to time small quantities of
HNO3 were added to maintain the volume. Refluxing of the sample continued until the sample was a light
amber color (two to four hours). The watchglass was then removed and the sample allowed to evaporate to
near dryness.
The residue was taken up in several ml of (1:1) HC1, warmed, and then filtered through a glass-fiber
filler. The filtrate was collected in a 20-ml centrifuge tube and insoluble silicates were removed by centrifuging
(2500 RPM) for 15 to 20 minutes. The supernatant was decanted into a 10-ml volumetric flask and the
precipitate rinsed with several ml of (1:1) HC1. The rinsings were added to the volumetric flask and it was
made to 10-ml with deionized water. This solution was used to analyze for Cd, Cu, Pb, and Zn as detailed
later.
Preparation of the mixed liquor samples for Hg analysis was according to the recommended EPA
method using the manual cold vapor technique^28'.
A 100-ml aliquot of the sample was pipetted into a 300-ml BOD bottle and 5 ml of 0.5 NH2SO4 and 2.5
ml of 16N HNO3 was added to digest the sample. Sufficient KMnO4 (20%) solution was added to each sample
to ensure a purple-colored solution. Complete oxidation of organic mercurials was accomplished by adding 8-
ml K2SO< (5%) solution and heating the samples in a water bath (95 °C) for two hours. The sample bottles
were allowed to cool and 6 ml of NH2OH. NaCl (12% v/w) solution was added to reduce the excess KMnO4.
The samples were then moved to the Hg analyzer where Hg concentrations were determined as described
below.
54
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Atomic absorption analyses of the mixed liquor sample digestates were performed according to the
recommended EPA methods*28' and the AAS instrument manufacturers' procedures*29'.
The mixed liquor samples were analyzed for Cd, Pb, and Hg. Both flame AAS and flameless AAS were
used to determine the Cd and Pb concentrations while the cold-vapor AAS technique was used for Hg
analysis.
Mixed liquor samples collected in February 1976 required the more sensitive flameless AAS technique
because of the relatively low Cd and Pb concentrations present. The October 1976 samples were much higher
in Cd and Pb and the flame AAS technique was required. Mercury also followed the same trend as Cd and Pb
but the linear range of the cold-vapor AAS technique was sufficient to measure both sampling periods.
Cadmium analyses (flame) of the October 1976 samples required a 1:1 dilution with deionized water to
bring the Cd concentration within the linear range of the instrument. Cadmium and Pb absorption were
measured at 228.8 nm and 283.3 nm, respectively, for both flame and flameless AAS techniques.
Detection limit for Cd by the flame technique was 0.006 mg// and by the flameless was 0.0002 mg//. The
detection limit for Pb by flame was 0.09 mg//and by the flameless technique was 0.003 mg//. Detection limit
is defined as that concentration of analyte which will give a response 2X the standard deviation of a series of
spiked samples which are detectable above the background noise.
Each mixed liquor sample was prepared and analyzed in duplicate for both Cd and Pb. The average
relative standard deviation (RSD) for duplicate samples by the flame AAS technique was 3.8% for Cd and
13.5% for Pb. The average RSD for duplicates by the flameless AAS method was 40.6% for Cd and over
50% for Pb. The high RSD for Cd and Pb by the flameless method results from the very low concentrations
of these analytes in the samples.
Recovery of Pb spiked mixed liquor samples ranged from 93% for flame AAS to 87% for flameless
AAS. Recoveries of Cd spiked mixed liquor samples ranged from 89% for flame AAS to 120% for flameless
AAS.
Mercury analyses of the digested mixed liquor samples were completed by adding 5 ml of 10% SnCh
solution to the BOD bottles and immediately connecting the bottle to the analytical train. The reduced Hg was
swept into the absorption cell and the Hg absorption recorded at 253.7 nm.
The detection limit for Hg by this procedure was 0.0025 mg//and was limited only by the reagent blank
Hg concentration.
Recovery of Hg spiked samples averaged 103% and the average RSD for duplicates at the 0.47 mg//con-
centration level was 7.9%.
Particulate Air Samples-
Cadmium, Cu, Pb and Zn were determined in air paniculate samples by the procedure of Thompson,
Morgan, and Purdue'30'.
The 20.3 x 25.4-cm glass-fiber filter was removed from its polyethylene bag and placed on a
polyethylene sheet. A plexiglas template and stainless steel surgical knife were used to cut a 2.5 x 20.3-cm
strip from the filter. The remainder of the filter was returned to its polyethylene bag.
The strip of filter removed was carefully cut into 1-cm lengths and placed into a 2.5 x 8.5-cm Pyrex ex-
traction thimble using Teflon-coated forceps. Eighty milliliters of digestion acid (16 ml of redistilled HC1 and
64 ml of redistilled HNO3) was added to the specially constructed boiling flask (Figure 9) and the extraction
thimble carefully lowered into its neck. The flask was then connected to the condenser unit and heat was ap-
55
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Dust Cover
Water
34/45 Groundglass Joint
A Ahlin-type condenser
B Extraction vessel with coarse glass disk
C Specially constructed boiling flask
D Hot plate
Figure 9. Apparatus for the acid digestion of air samples.
56
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plied. Once the acids began to reflux and wash over the filter strips, the temperature was adjusted such that a
continuous refluxing occurred. Refluxing was continued for three hours before the heat was removed and the
digestion acids allowed to cool. Several milliliters of 0.1 N HNO3 were poured through the top of the con-
denser and allowed to drain into the boiling flask.
The boiling flask was removed from the condenser unit and the extraction thimble removed. The boiling
flask was placed on a hot plate (300 °C) and a flow of N2 added to assist in evaporating the digestion acids.
Once the volume was reduced to several milliliters (never to dryness) the flask was removed from the hot
plate, cooled, and quantitatively transferred to a graduated centrifuge tube (15-ml). The sample volume was
made to 5.0 ml with deionized water before the sample was centrifuged for 30 minutes at 2000 RPM. A por-
tion of the filtrate was decanted into a polyethylene vial (5-ml) taking care not to disturb the precipitate.
Cadmium analysis was done on this filtrate by aspirating into an air-acetylene flame (AAS). A 1-ml ali-
quot of this filtrate was pipetted into a 10-ml volumetric flask and made to volume with deionized water. This
dilute (1:10) solution was used for Cu and Pb analysis using flame AAS.
The detection limit for each metal, based upon a 2000 mj air sample, was: Cd = 0.0003 ftg/m3, Cu =
0.004 /ig/m3, Pb = 0.045 /ig/m3, and Zn = 0.03 /tg/m3.
With each day's analysis, a series of spiked filter blanks was analyzed. The average recovery of these
spikes for Cd (0.5 /tg), Cu (5.0 /*g), Pb (250 /*g), and Zn (250 /ig) was 100%, 91%, 96%, and 87%, respective-
ly.
Mercury concentrations in the ambient air were measured by the charcoal absorption procedure of Scar-
ingelli, Puzak, Bennet, and Denny(48). This procedure was used on all sampling periods except the last
(October 1976). The amalgamation of Hg on silver wool by the procedure of Wroblewski, Spittler, and Har-
rison*3 ') was used for the last sampling period.
This change in the air-Hg methodology resulted from continuous problems which plagued the charcoal
absorption procedure. These problems were (1) difficulties in cleaning the charcoal, and (2) contamination.
Each time the charcoal tubes were fired for cleaning or sample analysis, some unknown compound
would bleed off the charcoal and mask the UV absorption of Hg at 253.7 nm. It would take several firings of
each tube to reduce this signal to an acceptable level. But after several minutes, refiring of the tube would
again give this masking signal.
Also the charcoal tubes would pick up Hg and/or some unknown compound from the atmosphere no
matter what precautions were taken to protect the tubes.
The detection limit of Hg spiked on the charcoal tubes was 0.024 ftg/m3 based on an average air volume
of 0.985 m3. These parameters were determined under the best of conditions and have limited meaning in rela-
tionship to the actual samples collected and analyzed due to the problems mentioned earlier.
The amalgamation of Hg on silver wool proved to be a very convenient procedure for determining Hg-air
concentrations.
The system used was that given by Wroblewski, Spittler and Harrison^. The tubes containing the Ag-
wool were fired to remove any traces of Hg absorbed on the Ag-wool. The tubes were immediately sealed for
shipment to the sampling site.
Several of the tubes were shipped to the sampling site and returned unused as checks on contamination
during shipment. None of these tubes gave a Hg signal when fired.
57
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The detection limit for Hg with the Ag-wool system was 0.0018 /ig/m3 based upon an average volume of
sampled air (0.2839 m3). Recovery of Hg spikes added to the Ag-wool was 100%.
Soil Samples-
Soil samples were prepared and analyzed by a modification of the methods used by Smith and
Windom'32' and Chester and Hughes^33) for sediment samples.
Several grams of the soil sample were dried overnight at 60 °C and the dried sample was then ground in a
mortar and pestle. A 5-gram portion of the pulverized sample was weighed into a 250-ml Erlenmeyer flask
and 50 ml of leaching solution [hydroxylamine hydrochloride:acetic acid (3:7)] added. The samples were then
shaken overnight (15 hours) on a mechanical shaker. The solution was then filtered through a glass-fiber filter
and the filtrate collected in a polyethylene bottle. All metals were determined by aspiration of this solution in-
to an air-acetylene flame (A AS).
Detection limits for the metals were: Cd = 0.01 /tg/g, Cu = 0.12 /ig/g, Pb = 0.66 /ig/g, and Zn = 1.8
-
Average recoveries were based on spiked soil samples carried through the leaching procedure: Cd (0.5
= 96.8% ;Cu (2 /ig/g) = 91.7%; Pb(5/tg/g) = 91.8%; Zn (20 /ig/g) = 106%.
Precision (RSD) based upon repetitive determinations (n = 6) of spiked samples was: Cd (0.20 /ig/g) =
1. 6%, Cu (0.83 /ig/g) = 7.4%, Pb (22 /ig/g) = 7.5%, Zn (8.4 /tg/g) = 10.9%.
Mercury analysis of soil samples was from a modification of the recommended EPA procedure for H% in
sediments(28).
One gram of the dried and pulverized soil (described above) was weighed into a 2.5 x 13.0-cm digestion
tube. Three milliliters of digestion acid HC1O4:HNO3(5:1) and 1 ml of KMnCX (5%) were carefully added to
the sample and it was gently shaken to mix the reagents. The sample was then placed into a water bath (75 °C)
overnight (15 hours). After allowing the sample to cool, 1 ml of hydroxylamine hydrochloride (50%) solution
was added to the sample and mixed. From this point the sample was analyzed by the conventional cold-vapor
technique for Hg after adding SnCb to reduce the Hg to its elemental state.
Quantitation for Hg was achieved by analyzing a series of standards prepared by this procedure and con-
structing an analytical graph (peak height vs concentration). The unknown samples were then quantitated
from this analytical curve.
Detection limit for Hg using this procedure was 0.006 /ig/g with an average recovery of 75%.
Precision, as RSD, for replicate (n = 7) determinations on a soil sample spiked at 0.04 /ig/g was 7.5%. A
sample spiked at 0. 10 /ig/g gave an average recovery of 85.3%.
Water Samples-
Water samples were prepared by the EPA method for total metals(28) and analyzed by the flameless AAS
technique for Cd, Cu, and Pb. Zn was analyzed by the flame AAS method.
Detection limits for the elements were: Cd = 0.0007 mg/i, Cu = 0.0032 mg/j£, Pb = 0.003 mg//, and
Zn = 0.02mg/£.
Each metal was quantitated by spiking one of the water samples at three different concentration levels
with the analyte metals and making an analytical curve (peak height vs. concentration) using the H-P 9810A.
The other water samples were then quantitated for each particular metal using the appropriate analytical
curve.
58
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The RSD of five replicate samples spiked with each metal were: Cd (0.002 mg//) = 15.9%, Cu (0.009
mg//= 18.3%,Pb(0.009mg//) = 14.8%, and Zn (0.08 mg/$ = 13.6%.
Average recovery of spiked samples was: Cd = 101%, Cu = 97%, Pb = 92%,andZn = 91%.
Water samples were analyzed for Hg by the recommended EPA method of manual cold-vapor
The detection limit was 0.00002 mg/^-and the RSD of four replicate samples at the 0.00006 mg/^ concen-
tration level was 15.2%. Average recovery of Hg spiked (0.00005 mg//) water samples was 80%.
Microbiological Analysis of Clinical1 Specimens
Bacteriology-
Feces Samp/es-Bacteriological examinations of feces were performed in accordance with the standard
procedures and techniques which have been established for isolation and identification of microorganisms by
clinical diagnostic laboratories and by procedures standard for this laboratory. Techniques follow generally
those described in the references cited above. Feces samples were inoculated onto S-S Agar, Endo Agar, Eosin
Methylene Blue Agar, and MacConkey Agar Plates for primary isolation. Cultural morphology, biochemical
reactions, microscopic appearance, and serologic testing were employed, as required.
Throat Swab Samp/es-Bacteriological examinations of throat swabs were performed in the same manner
as the feces samples. Throat swabs were inoculated primarily onto Blood Agar Plates.
Sputum Spec/mens-Sputum Specimens were inoculated onto Loewenstein-Jensen Medium and Mid-
dlebrook 7H-10 Medium in screw cap tubes and incubated in a CO2 incubator.
Virology-
Feces Samp/es-Isolation and identification of viruses from feces were performed by standard
procedures(26) and by procedures of the NIH/WHO Regional Reference Center for Simian Viruses at SFRE.
Monolayer cell cultures of primary baboon kidney and cultures of a continuous cell line (WI-38) were
employed for demonstration of cytopathology.
Throat Swab Samples-Virologic examinations of throat swabs were performed by the methods used for
feces and throat swabs. Primary isolation was performed on primary baboon kidney cells and WI-38 cell
monolayers.
Viral Serology-Antibody studies for experience with viral agents were performed by methods given in the
references above. The specific test system employed — hemagglutination-inhibition or serum
neutralization — is designated on the respective tables.
Parasitology on Feces Samples-
Samples collected by the human subjects were processed at the field laboratory near the study area. This
processing consisted of placing a small amount of feces in a 15-ml capacity screw cap vial containing 9 ml of
merthiolate-formalin. One ml of iodine was added to this solution just prior to the feces samples.
Analysis of the samples took place at the San Antonio laboratories. The merthiolate-iodine-formalin
concentration technique (MIFC) is that method originally proposed by Blagg, et al.^34^. This simple technique
consists of shaking the vial with NIF to insure a homogeneous solution. Two layers of gauze strain the solu-
tion as it is poured into a centrifuge tube. One to three ml of ether was added and the tube was vortexed for 10
seconds. The tube was then centrifuged at 2500 RPM for two minutes. The supernatant was discarded and the
sediment was examined under medium- and high-power (100X) magnification. The results of the analysis are
reported either as no parasites seen or, if any parasites are found, as identified.
59
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Chemical Analysis of Human Tissue Samples
Trace Metal Analysis--
Instrumentation-lnstrumentation and reagents used for the trace metal determinations in human tissues
were identical to those listed above.
Blood-Venous blood samples were analyzed for Cd and Pb by a modification of the atomic absorption
methods of Hwang, Ullucci and Mokeler^34^ Hauser, Himmer and Kent^35', and Mitchell, Ryan and
Aldous(36).
A 2-ml aliquot of the thawed blood sample was pipetted into a 5-ml screw-cap extraction tube and 0.5 ml
of Trisma buffer solution (pH 7.0) added. Next, 0.5 ml of a chelating-hemolyzing solution consisting of 2%
(w/v) ammonium pyrrolidinedithiocarbamate (APDC) in a 2% (v/v) solution of Triton X-100 was added to
the extraction tube. The tube was capped and vigorously shaken for several minutes then allowed to stand for
10 to 15 minutes to ensure complete hemolysis of the blood. To extract the Cd and Pb chelates, 0.5 ml of
M1BK was added to the extraction tube and shaken for five minutes. The sample was then centrifuged to 2500
RPM for 10 minutes and the MIBK layer (top) removed to a 2-dram glass vial.
Cadmium and Pb determinations were made on this extract by pipetting 5- to 10-/il aliquots into the
preprogrammed HGA-2000 graphite furnace and measuring the absorption at 228.8 nm (Cd) and 283.3 nm
(Pb).
Quantitation of the blood extracts were by the "method of additions" using whole blood purchased
from a local blood bank. These spiked blood standards were prepared and analyzed by the procedure outlined
above. Analytical curves were prepared from these spiked blood standards using the H-P 9810A.
This procedure for blood-Pb analysis has been verified by participation in the Center for Disease Con-
trol's Blood-Pb Proficiency Testing Program.
Detection limits for blood-Cd and blood-Pb by this methodology are 0.014/tg/100 ml and 1.4/ig/100 ml,
respectively.
The RSD for blood-Cd spiked at 0.12 /ig/100 ml was 5.9% (n = 6) and for blood-Pb spiked at 7.6 /ig/100
ml was 13.1%(n = 6).
Average recovery of spiked blood for Cd was 98.5%. Recovery of blood-Pb was based on CDC bovine
blood samples and averaged 102.1 %.
Copper and zinc in whole blood samples were determined by a modified Matousek and Steven^ pro-
cedure.
A 100-/il aliquot of the thawed blood sample was pipetted into a 5-ml polyethylene vial. For Cu deter-
minations, the blood aliquot was diluted 1:9 with deionized water and mixed well. Zinc determinations were
made by diluting the blood aliquot 1:39 with deionized water and thoroughly mixing.
Both the Cu and Zn were determined by aspirating the diluted blood into an air-acetylene flame and
measuring the absorption at 324.7 nm (Cu) and 213.9 nm (Zn).
The "method of additions" was used to quantitate both Cu and Zn. The procedure was the same as that
described above for Cd and Pb. Detection limits for blood-Cu was 16.5 /tg/100 ml and for blood-Zn was 34.6
/ig/100 ml. A spiked blood sample containing 105 /ig/100 ml Cu had an RSD of 8.6% (n = 6) and another
sample containing 577/ig/100 ml Zn had an RSD of 2.2% (n = 7).
60
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Average recoveries of spiked blood samples for Cu and Zn were 99.8% and 103.1%, respectively.
Blood-Hg determinations were by the cold-vapor method of Hatch and Ott<37) and Skare(38).
A 1-ml aliquot of the thawed whole blood was pipetted into a 2.5 x 13.0-cm digestion tube with a 24/40
ground glass joint. Three ml of HCKX:HNO3(5:1) was added and the blood was allowed to digest overnight
in a water bath at 75 °C. After the samples had returned to room temperature, 3 ml of deionized water and 1
ml of hydroxylamine hydrochloride solution (50%) was added. The samples were mixed on a Vortex shaker.
Next, 5 ml of 10% SnCl solution was pipetted into the digestion tube and it was immediately connected to the
cold-vapor analytical train. The digestion tube was continuously shaken as the flow of N2 bubbled through
the sample and the released Hg vapor absorption was measured at 253.7 nm.
Quantitation of blood-Hg samples was by the "method of additions" as described for blood-Cd and Pb.
The detection limits of blood-Hg by this procedure was 0.1 /ig/100 ml and a spiked blood sample at 0.45
Aig/100 ml had an RSD of 7.8% (n = 7).
Feces-Feces samples were prepared for Cd, Cu, Pb, and Zn analyses by the following methodology: A
1.5-gram aliquot of thoroughly mixed feces samples was weighed into a Teflon bomb^9\ Five ml of concen-
trated HNO3 was added and the bomb sealed. The bomb was placed on a hot plate (125-150 °C) for two hours
to digest the feces and then allowed to cool to room temperature before opening. The digested feces solution
was filtered through a glass-fiber filter and the filtrate collected in a 10-ml volumetric flask. Several milliliters
of 0.1 N HNO3 was used to rinse the bomb and filter. Deionized water was used to fill the volumetric flask to
10ml.
The method of Skare^38^ was used to prepare the feces samples for Hg analysis. A 1-gram portion of the
mixed feces sample was weighed into a 300-ml BOD bottle and 10 ml of concentrated H2SCX and 2 ml of con-
centrated HNO3 added. The bottles were protected from dust contamination and allowed to digest overnight.
Next, between 15 and 45 ml of 30% KMnCX solution was added and the sample was placed on a steam bath
for 30 to 90 minutes. Approximately 100 ml of deionized water was added followed by 6 ml of 12% hydrox-
ylamine hydrochloride. The sample was ready for Hg analysis as described below.
The diluted feces digest solution (10 ml) was used for the determination of Cd, Cu, Pb, and Zn.
Cadmium and copper were determined by aspirating the digest solution directly ii to an air-acetylene
flame and measuring the absorption at 228.8 nm (Cd) and 324.7 nm (Cu).
One milliliter of the 10-ml digest solution was diluted to 50 ml with deionized water and this diluted
(1:500) digest was aspirated into an air-acetylene flame for Zn analysis at 213.9 nm.
Lead was analyzed by injecting 10-/il aliquots of the digested feces solution (10 ml) into a preprogrammed
HGA-2000 graphite furnace and measuring the absorption at 283.3 nm.
Quantitation of Cd, Cu, Pb, and Zn was by making spiked standard solutions at the same acid concen-
tration here as the digested samples and developing an analytical curve (concentration vs. peak height) with
theH-P9810A.
Detection limit for Cd and Cu in feces was 0.014 /*g/g and 1.5 /*g/g, respectively, based upon wet
weights. The detection limits for Pb and Zn in feces were 0.10 /tg/g and 7.3 /tg/g(wet weight), respectively.
Recovery of analytes was determined from spiked feces samples carried through the preparation and
analysis procedures. Cadmium recoveries averaged 94.3% while Cu recoveries averaged 95.6%. Recoveries of
Pb and Zn averaged 104.2% and 99.0%, respectively.
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Feces samples spiked at 0.15/ig/g Cd, and 11.5 /ig/g Cu had RSD of 15.7% (n = 11) and 9.3% (n = 5),
respectively. While feces spiked with Pb at 0.64 /ig/g had an RSD of 23.4% (n = 5) and spiked with Zn at 77
/ig/g had an RSD of 1.4% (n = 3).
//o/>--Preparation of the hair samples for trace metal analyses was patterned after the methods of Ham-
mer, et al.*40) and Harrison, et al.'41^
Hair samples as received were cut into approximate 1-cm lengths using stainless steel surgical scissors.
The cut hair was collected on a clean polyethylene sheet and transferred to a 250-ml Erlenmeyer flask. Then
200 ml of sodium laural sulfate solution (0.04% w/v) was added and the flask was placed on a rotating shaker
for one hour. The surfactant solution was decanted off and the hair rinsed twice with 200 ml of deionized
water. Both rinses were also decanted off. A third 200-ml portion of deionized water was added to suspend
the hair sample and immediately poured into a 150-ml glass-fritted filter (coarse). The water was removed
under vacuum filtration and another deionized water rinse followed.
After removal of this last rinse, 100 ml of isopropyl alcohol was added to the sample and it was allowed
to soak for approximately five minutes with occasional stirring. The alcohol wash was removed by vacuum
filtering and the hair was again rinsed twice with deionized water.
Once most of the water had been removed, the hair sample was transferred to a 150-ml polyethylene
beaker and covered with a piece of loose-woven paper to prevent dust contamination. The beaker was then
placed in a drying oven (110°C) for one to two hours. The beaker was removed from the oven and the washed
and dried hair was ready for digestion.
A 1-g portion of the washed and dried hair sample was weighed into a 250-ml Vycor beaker using Teflon-
coated forceps to transfer the hair. Digestion of the hair was accomplished by adding 10 ml of
HNOj:HClO4(l:l) to the sample and allowing it to stand overnight. The sample was then placed on a hot
plate (200 °C) and a flow of N2 added to reduce the volume. This operation was performed in a stainless steel
hood. The sample volume was reduced to 1 to 2 ml or until dense white fumes were given off. Under no cir-
cumstances was the sample allowed to go to dryness during this step.
The beaker was removed from the hot plate and allowed to cool before quantitatively transferring the re-
maining digest solution to a 10-ml volumetric flask. Rinsing off the digestion beaker and filling of the
volumetric flask to 10 ml was done with IN HNO3. The diluted hair digest was then placed in a polyethylene
bottle and stored under refrigeration (4°C) until analyzed.
The diluted hair digest was allowed to reach room temperature before aspirating into an air-acetylene
flame for Cd, Cu, Pb, and Zn analyses. Cadmium absorption was measured at 228.8 nm, Cu at 324.7 nm, Pb
at 283.3 nm, and Zn at 213.9 nm.
A composite hair sample was made from a large quantity of hair obtained from a local barber shop. This
composite hair sample was washed according to the method above and spiked with all metals being determin-
ed prior to digesting. These spiked, composite hair samples were analyzed daily to quantitate the hair samples
(method of additions). Calculation of analytical curves from peak heights vs. concentration were performed
on the H-P 9810A as described earlier.
Detection limits were: for Cd in hair 0.13 /ig/g; for Cu 1.35 /ig/g; for PbO.87 /ig/g; and for Zn 15.1 /ig/g.
Spiked composite hair at 0.90 /ig/g Cd gave an RSD of 8.9% (n = 21). Hair spiked at 19.4 /tg/g Cu gave
an RSD of 9.2% (n = 4) while at 14.0/ig/g Pb gave an RSD of 15.1% (n = 17). Hair spiked with Zn at 164
/ig/g had an RSD of 2.2% (n = 5).
62
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Recoveries of spiked composite hair samples were: Cd = 97.3%,Cu = 106.5%, Pb = 83.0%, and Zn =
101.5%.
Concentration of Hg in hair was determined in the following manner. A 0.1-g portion of the washed and
dried hair was weighed into a 2.5 x 13.0-cm digestion tube with a 24/40 ground glass joint. Five ml of
HC1O4:HNO3(5:1) was added. The HC1O4 was ultrapure grade. Next, a 1-ml aliquot of 5% KMnO4 solution
was added and the digestion tube was placed in a water bath (75 °C) for 15 to 20 minutes. This operation was
performed in a stainless steel fume hood. After the sample was removed from the water bath, 10 ml of
deionized water and 1 ml of hydroxylamine hydrochloride solution (50%) was added. The sample was
thoroughly mixed and 5 ml of a 10% SnCl2 solution added. The digestion tube was immediately connected to
the cold-vapor analytical train. The Hg° vapor released from the sample was swept into the absorption cell
and Hg absorption was measured at 253.7 nm.
Mercury in hair was quantitated by processing a series of aqueous spiked standards along with the
unknown hair samples. Analytical curves were plotted with the H-P 9810A using peak height vs. concentra-
tion of the aqueous spiked standards.
Using this procedure and 0. Ig of hair, the detection limit for Hg in hair was 0; 17 /tg/gm of dry hair.
Average recovery of Hg-spiked hair samples was 79.3% and a spiked sample at 0.35 ^g/g gave an RSD of
9.1% (n = 7).
L/A//7e-Determination of Cd, Cu, and Pb in urine was patterned after the extraction procedure of Kubasik
and Volosin^ and Parker, Humoller and Mahler'43\
A 10-ml aliquot of acidified urine was pipetted into a 10-ml screw-cap extraction tube and its pH ad-
justed to 7.0 with concentrated NH4OH. One ml of Trisma Buffer (pH 7.0, 0.5M) solution was added. The
sample was capped and vigorously shaken for one minute to thoroughly mix the reagents. Following this
shaking, the pH of the sample was rechecked.
A 1-ml portion of a 2% (v/v) Triton X-100 solution containing 2% (w/v) APDC was added to the buf-
fered urine sample and the sample was thoroughly mixed. Extraction of the chelated metals was accomplished
by adding 1 ml of MIBK (water saturated) and vigorously shaking the sample for one minute. Centrifuging at
2500 RPM for 10 minutes separated the aqueous and MIBK layers and the latter was removed to a small glass
vial for AAS analysis.
Cadmium, Cu, and Pb determinations were immediately performed on this urine extract using the
HGA-2000 graphite furnace. Cadmium was determined at 228.8 nm, Cu at 324.7 nm, and Pb at 283.3 nm.
A composite urine sample was collected in our laboratories and spiked with each of the analyte metals at
three different concentration levels. This composite urine was used to quantitate the unknown urine samples
by the "method of additions." Peak height vs. concentration regression curves (analytical curves) were
calculated for each of the three metals using the H-P 9810A calculator. The unknown urine samples' metal
concentrations were determined from these curves.
A Cd spiked urine sample at 0.234 /tg// had an RSD of 12.9% (n = 6) while Cu-spiked urine at 4.3 /ig/£
had an RSD of 16.8% (n = 9). Lead-spiked urine at 7.8 /tg/Ahad an RSD of 16.0% (n = 8).
Recoveries of Cd, Cu, and Pb averaged 101.8%, 108.8%, and 107.7%, respectively, based on spiked
urine samples.
Zinc determinations in urine were based on the ^ method of Parker, Humoller and Mahler.
63
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An aliquot of the acidified urine was diluted 1:1 with deionized water and aspirated directly into an air-
acetylene flame. Because of the nature of the diluted urine sample, a high-solids burner head was used for this
analysis. Zinc absorption was measured at 213.9 nm.
Quantitation of urine-Zn was the same as described above for Cd, Cu, and Pb. An Zn-spiked urine sam-
ple at 340 /tg/^ had an RSD of 3.9% (n = 4) and a recovery of 100.9%.
The method of Skare*38* was used to measure urine-Hg levels.
A 1-ml aliquot of urine sample was pipetted into a 2.5 x 13.0-cm digestion tube with a 24/40 ground
glass joint and 0.2 ml of concentrated H2SO4 added. The samples were placed in a water bath to cool and
allowed to stand approximately 10 minutes with occasional mixing. Next, a 2-ml portion of 6% KMnO4 solu-
tion was carefully added and mixed. The digestion tube was protected from dust contamination and allowed
to stand overnight at room temperature.
The following day, the excess KMnCX was reduced by adding 0.3 ml of hydroxylamine hydrochloride
solution (20%). The sample was then removed to the Hg analyzer and 5 ml of 10% SnCh solution added. The
digestion tube was immediately connected to the cold-vapor analytical train and Hg absorption was measured
at 213. 9 nm.
A few of the urine samples had heavy precipitates when received for analysis. These samples were
thoroughly mixed and a 10-ml aliquot removed for Hg analysis. The same procedure outlined above was
followed only 10X more reagent volumes were used.
Quantitation of urine-Hg was by processing aqueous spiked standards along with the samples and
calculating an analytical curve (peak height vs concentration) with the H-P 9810A calculator. Concentration
of unknown urine samples was determined from this curve.
Detection limit for urine-Hg by this procedure was 1.14 ^g// with a recovery of 102.5% based on spiked
urine samples. A urine sample spiked at 7.83 /tg// Hg had an RSD of 7.3% (n = 7).
Hematocrit Value of Blood Samples-
Hematocrits were determined on the blood samples at the time of collection. A Clay-Adams hematocrit
centrifuge was used to spin the collected capillary blood tubes and the hematocrit value was read directly from
the calibration graph on the centrifuge.
DATA FLOW AND PROCESSING
Identification and Integrity of Sample Data
The data utilized in this study came from a variety of sources: environmental samples from the vicinity of
the sewage treatment plant, household health surveys of residents within 5 kilometers of the plant, and human
subjects from these households who volunteered to participate in the study. The human subjects periodically
provided various clinical specimens and health questionnaire information, as depicted in Table 1 in Section 1 .
To develop valid findings from this multitude of sample analysis and questionnaire data, it is essential
that the identity and integrity of the data be preserved. Accordingly, an integrated data system involving a
unique sample code, computer-generated sample labels, keypunched questionnaire forms, and analytical data
reporting forms was implemented. This system insured that each sample was uniquely identified (participant
ID or sample location, medium, analysis, sampling period) and that this identity was transmitted with the
analytical values from sample collection through processing, storage, preparation, analysis, reporting, and
statistical analysis.
64
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We found that unique sample identification could be readily obtained by developing unique identifiers
for the various aspects of a sample: the participant or location sampled, the medium sampled, the type of
analysis to be performed, the sampling period, and necessary subsampling information. Distinct codes were
assigned to each aspect. Volunteers were assigned consecutive numbers so that each selected participant had a
unique ID number. Unique identifiers were also devised for each sample medium (air, soil, water, feces,
blood, throat swab, etc.), each type of sample analysis (bacteria, viruses, trace metals, etc.), each sampling
period, each sample location (for the environmental samples), and for each subsample. These identifiers were
combined as a unique 15 digit code giving the history, contents, and planned analysis of each sample, and
distinguishing this study's samples from those of other concurrent epidemiological studies. The composition
and interpretation of this sample code is given in Table 7 . A typical sample code is 0478TBCH2--4007. This
code identifies the throat swab (T) for participant 478 from sampling Period 2 at Chicago (CH) on this study
(4007); it is to be analyzed for bacteria (B).
Sample Labels
Sample labels were used to affix the sample code to every bottle containing the sample or a portion of the
sample. Appearing on the sample label were both the sample code (in the lower left hand corner) and several
printed words. The printed words gave enough information to the field personnel to properly process and
distribute the sample without needing to interpret the sample code. Thus the sample labels facilitated correct
sample processing and distribution while preserving the sample's identity. Typical sample labels are given in
Figure 10 . The last sample label in Figure 10 , for example, would be placed on the bottle containing the por-
tion of the BHI fluid from the air sample obtained by high volume sampler 1 on run 20 during Period 5 that is
to be analyzed for enteroviruses (VIRUS) by the UTSA laboratory.
Special computer programs were written to generate the proper types and numbers of sample labels. The
programs also sorted the labels so that they were printed in the sequence that was most convenient for their
application to the proper sample containers before and during sample collection and processing in the field.
For some sample media, the collected sample had to be split into several portions at the on-site field
laboratory, because several analyses, requiring either differing preservatives or different laboratories, were to
be performed. The proper number and type of processing labels were also generated for the bottles containing
these processed sample portions. The collection sample labels contained both the participant's name and his
identification number to minimize mistakes during sample collection. However, the processing sample labels
contained only the identification number to maintain confidentiality.
Data Forms and Reporting System
An efficient data reporting system was developed. The questionnaires for the household health survey,
for participant recruitment, and for current health status were designed for direct keypunching after minimal
coding during a manual scan. Special forms for reporting field and laboratory data were developed in col-
laboration with the field and laboratory supervisors for direct coding in a suitable data entry format. Coded
examples of the following data forms are presented in Appendix G.
— Aerosol Run Report on Meteorology and Sampling
~ Configuration
- UTSA-CART Aerosol Run Analysis Report
- Household Health Survey
- Health Survey Participant Questionnaire
- Current Health Status Questionnaire
- Metal Analysis Coding Form
-- Pathogen Analysis Coding Form
- Pathogen Analysis Data Form
As these coded examples illustrate, each laboratory result is identified by including the relevant parts of the
65
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TABLE 7. THE FIFTEEN-DIGIT SAMPLE CODE
Columns
1-4
Variable
Participant or location identification:
• For clinical specimens: participant
ID number
Codes
• For environmental samples: location
- Soil
- Water
- Aeration basin mixed liquor
- Air
- High volume microbiological air
Sampler number (Col. 1, 2)
Run number (Col. 3, 4)
Sample medium
Sample analysis
0001 - 0654
7-8 Sampling site
9 Sampling period
10-11 Detailed time or location
12-15 Project number
S001 - S050
W001 - W010
R001 - R010
UOUT, U-IN, 50,
500, 1600, 5000
01 - 12
01 - 20
A - Air
B - Blood
F - Feces
H - Hair (long)
I - Hair (short)
R - Aeration basin
S - Soil
T - Throat swab
U - Urine
W - Water
Z - Sputum
A - Antibodies
B - Bacteria
C - Coliphage
H - Mercury
M - Trace metals
P - Parasites
V - Viruses
(blank) - Several analyses
CH - Chicago
1 - 5
Miscellaneous
numerical
4007
66
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BLOOD
1BACH55 1007
BLOOD
FIFTH SAMPLE
FECES
•fFBCHSB 4007
FECES
FIFTH SAMPLE
HAIR
SPUTUM
THROAT SWAB
URINE
HJ2Z CH11 HUU7
5TAC1&5
fl^UMCHil HUU7
Figure 10. Typical sample labels.
MAIN-LONG
FIFTH
SPUTUM
FIRST SAMPLI:
1'HKOAT fcwB
FIFTH SAhi-Lt
UHlNt
F1KS1 SAMPIC
61
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SEWAGE
BACTERIA
SEWAGE
COLIPHAGE
AIR
BACTERIA
AIR
MERCURY
AIR
TRACE METALS
AIR
VIRUS
SAMPLER 2 SEWAGE
RUN i BACTERIA
2 ISdCHS 2tOU? AuUA
SAMPLE* i SEWAGE
RUN lu COLIPHAGE
110SCCH5 IfOO? UTSA
SAMPLER 2 AIR
RUN 3 BACTEKIA
2 3ABCHS 11007 AQUA
SAMPLER I, AIR
RUN i MERCUKY
b lAMCHS ffOO?
SAMPLER e AIR
RUN i METALS
2 1AMCH5 1H007 SwRI
SAMPLER 8 AIR
RUN 10 VIRUS
810AVCH5 11007 UTSA
Figure 10. Typical sample labels (continued).
68
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sample code on the laboratory data form. When the entire set of data forms for a sampling period were com-
pleted, the laboratory or field supervisor transmitted them as a package for data processing to avoid problems
with incomplete data sets.
Participant and Household Survey Data Bases
The data reporting system was designed to permit construction of an automated participant data base by
utilizing the natural structure of this data. The records to comprise the data base were the participant ques-
tionnaire data, precise residential distance and direction information, the pathogen isolations and antibody
liters from the pathogen analysis coding form, the trace metal concentrations from the metal analysis coding
form, and the current health status questionnaire data. The records in this participant data base were to be
sorted primarily by participant ID, which appears in columns 1-4 of every record, secondarily by record type,
and thirdly by sampling period within record type. Statistical analyses would be performed by abstracting the
pertinent data from the data base. However, upon receipt of data sets from the early sampling periods, it
became evident that construction of the data base was not warranted. Because of the lags in reporting some
laboratory results, construction of the data base after receipt of all laboratory data would have delayed the
statistical analysis much beyond an acceptable project schedule. Furthermore, because of the limited nature
of the plant effect evident in the preliminary data inspection, many of the statistical analyses requiring an en-
compassing data base were found to be unnecessary. In fact, many of the statistical analyses could be per-
formed manually on the properly compared data. Consequently, a more limited participant data processing
approach tailored to the specific statistical analyses was implemented. The Pathogen Analysis Data Form,
which tabulates the laboratory results in the paired comparison form needed for the statistical analysis,
typifies the more limited ad hoc approach used for the participant data.
A household health survey data base was constructed because of the massive amount of questionnaire
data. The records in the household health survey data base were the eleven card images keypunched per ques-
tionnaire for each of more than 2200 households surveyed. This data base was sorted primarily by household
number (col. 1-4, with col. 1-2 identifying the sector and year) and secondarily by card number (col. 80). The
household health survey data base was stored on magnetic tape in 80 column card image form.
Computational Techniques
The automated computational techniques which were used to analyze the extensive data generated during
this project were chosen as being the most appropriate from numerous options that were available. SwRI cur-
rently maintains a special lease arrangement with the McDonnell Douglas (McAuto) computer facility in
Huntington Beach, California for use of their CDC Cyber 70/74 equipment. This system is accessed through
a CDC remote batch terminal at the Institute's Computer Laboratory. In addition, a Hewlett Packard 9810A
programmable calculator with a limited package of statistical routines was utilized when the desired analysis
was less involved and the quantity of data was sufficiently small to permit direct keyboard entry. The major
automated computational procedures used in this project are listed in Table 8.
The major computer programs written for this project were those to generate sample labels and those to
analyze the household health survey data base. The function of the sample label program is discussed
previously in this section. The programs HHSTAB, HHSURV, and TESTX2 read the household health
survey data base, tabulate the distributions of characteristics among households and the distributions of
illnesses, diseases, and symptoms among household member categories, and compute the survey analysis test
statistics.
GENERAL STATISTICAL METHODOLOGY
The specific research questions given in Section 1 have been addressed by applying suitable statistical
methods to the study data to make appropriate inferences (i.e., findings). The general statistical analysis
69
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TABLE 8. AUTOMATED COMPUTING PROCEDURES
Program Source
MAILLB SwRI
LABELS SwRI
WRTLBLS SwRI
PSTAT1 SwRI
DSTAT3 SwRI
DSTAT4 SwRI
ANOVA HP
CHISQ HP
BMDO1D BMD
BMDO4D BMD
BMDO9S BMD
BMDO8V BMD
PRESTO SwRI
HHSTAB SwRI
HHSURV SwRI
TESTX2 SwRI
Computer
Type System
Usage
New
New
New
New
New
New
New
Cyber 74
Cyber 74
Cyber 74
Existing HP9810A
HP9810A
HP9810A
Package HP9810A
Package HP9810A
Package Cyber 74
Package Cyber 74
Package Cyber 74
Package Cyber 74
Existing Cyber 74
New Cyber 74
Cyber 74
Prints labels with names and
addresses of human subjects
Prints human subject, soil, and
water sample labels
Prints air and aeration basin sam-
ple labels
Mean, standard deviation, co-
efficient of variation
Calculate arithmetic and geo-
metric means, standard devia-
tion for grouped data
Calculate pooled arithmetic and
geometric means, standard devia-
tion for several groups of data
Analysis of variance; one-way and
two-way without interaction
2 x k chi-square
Mean, standard deviation, stan-
dard error, with transgeneration
Single character distribution
Transformation and generation
from data deck
General ANOVA model
Stepwise multiple linear regression
Distribution of household and human
subject characteristics by incidence
of symptoms and diseases
Distribution of household and human
subject data by multiple cross-
classifications
Cyber 74 Calculate chi-square statistic
70
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methodology employed is depicted in Figure 11 . Each research question has been translated into an ap-
propriate hypothesis testing framework, and pertinent statistical methods have been identified to test the null
hypothesis against the alternative. The validity of each potential method's assumptions has been evaluated by
considering the nature of the data to be analyzed in order to select the optimal statistical, method. The data
have then been analyzed by the selected method to draw inferences relevant to the research questions. In those
cases where the primary hypothesis testing produced an outcome with possible health hazard implications, a
secondary hypothesis was also tested to fully satisfy the research question. In the environmental monitoring,
for example, if the primary analysis indicated the sewage treatment plant was a significant source of a hazar-
dous agent, a secondary analysis was conducted to determine whether the downwind residential levels of the
hazardous agent were also elevated.
The particular statistical methodology utilized in each study area is presented in the applicable part of
Section 6, Results. As Figure 11 indicates, each of these statistical methodologies share many common
characteristics. The following paragraphs present the common elements for each step displayed in Figure 11 .
The discussion includes general rules which have been employed throughout the statistical analyses, with the
exceptions noted in Section 6 for particular analyses.
Sensitive variables and suitable hypotheses have been formulated for each research question, taking the
experimental design of the study area into account. The appropriate variables and hypothesis for a research
question depend upon the nature of sewage treatment plant exposure which might be measured in the various
types of sampling conducted.
Most of the sampling, including the health survey questions, the clinical specimens for microbiological
and trace metal analyses, and the soil and water environmental monitoring, might detect the accumulated ef-
fects of relatively long-term exposure to the sewage treatment plant. For these cumulative exposure measures,
the appropriate variable and hypotheses involve comparing the difference in the levels of the exposure
measure between the operational sampling period and the baseline sampling period at the same time of year.
If the sewage treatment plant does have detrimental health effects, one would anticipate an increase in levels
in the operational period, especially very close to the sewage treatment plant. Our general statistical
methodology in such study areas has been to test the null hypothesis of no difference between the operational
and baseline levels against the two-sided alternatives of either a significant increase or a significant decrease.
Since there are many possible reasons for an operational period increase that are unrelated to sewage treat-
ment, variables haying a significant operational period increase have been further examined to ascertain
whether the increased levels are related to exposure to (usually, distance from) the sewage treatment plant.
The air sampling, on the other hand, detects very recent and brief exposure to the sewage treatment
plant. The proper variables and hypotheses for air sampling compare downwind levels with upwind levels for
operational period sampling runs. The null hypothesis of no difference is tested against two-sided alternatives
of a significant downwind increase or decrease. The closest downwind levels are compared against the upwind
levels to determine whether the sewage treatment plant is a significant source of the measured hazardous
agent. When the plant is a significant source, the residential downwind and upwind levels have been com-
pared to see if the plant remains a significant source in residential areas.
The appropriate general approach has been utilized to formulate each research question in a statistical
hypothesis testing framework. Two-sided tests of the null hypothesis have been employed to insure objectivity
and to be conservative in the identification of possible sewage treatment plant health effects. While a 5%
significance level has been used in the hypothesis testing, interpretation is based largely on the P value of the
test statistic. Thus test results in the .05
-------
STUDY
OBJECTIVE
1
FORMULATE
SENSITIVE VARIABLE
AND HYPOTHESES
I
IDENTIFY
PERTINENT
STATISTICAL
METHODS
NATURE OF DATA
(Validity of Method
Assumptions)
SELECT
STATISTICAL
METHOD
DATA
ANALYSIS
FIND
INGS
Figure 11. General statistical analysis methodology.
72
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The validity of the assumptions of each potential method about the nature of the data to be analyzed
have been considered in selecting the most appropriate method. Parametric methods make a distributional
assumption, and their validity depends upon the appropriateness of the assumed distribution. The more sen-
sitive parametric methods generally assume the data are normally distributed. However, logarithmic transfor-
mation of the data permits application of the same methods for lognormally distributed data, which fre-
quently occur in environmental and health monitoring.
Inspection using statistics such as the coefficient of variation (defined as the standard deviation divided
by the mean) has often been used to choose the best parametric method. The degree of invalidity of the
parametric method assumptions has been weighed against the inefficiency (loss of power in rejecting the null
hypothesis when it is false) of using the analogous non-parametric method to select the best statistical
method. Because of the importance of this study, we have generally taken the conservative approach of using
non-parametric methods when the disadvantages of both methods appeared to be equal. In some cases, both
methods have been applied to the data, and we have reported the consistency of their results.
The selected statistical method was then used to analyze the data and make our findings about the
research question. Some sets of data contained "less than" values, where the measurement was below the
analytical detection limit. When they constituted a small proportion of the sample, these less than values were
replaced by half the lower detection limit arid used in the data analysis. Similarly, occasional "greater than"
values, above the upper detection limit, were replaced by twice the upper limit and retained in the data
analysis. However, when the "less than" values occurred often enough to invalidate the method assumptions,
no statistical analysis was conducted. As mentioned above, the P value associated with each test result was
used to interpret the relative significance of the finding regarding the research question.
73
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SECTIONS
RESULTS
ENVIRONMENTAL MONITORING
Aeration Basin Samples
Table 9 reports the results of the UTSA microbial characterization of two large volume aeration basin
samples taken in Period 5. The table shows the standard analyses, bacterial screen, and the distribution ob-
tained from Enterobacteriaceae testing of the two grab samples. It should be noted that the levels of several
pathogenic and potentially pathogenic genera - Klebsiella, Mycobacteria and Staphylococcus - were fairly
high relative to the microbiological indicator levels.
Samples of the mixed liquor were taken from an aeration basin along with each high volume aerosol
sampling run and microbiological analyses for standard plate count, total coliform and fecal coliform were
performed. The samples from both Period 4 and Period 5 were analyzed for these three indicators; in addi-
tion, Period 5 samples were analyzed for coliphage. The microbiological concentrations obtained from these
analyses, conducted by Aqualab, are reported in Tables 10 and 11. Note that the sample concentrations ob-
tained from the second stage aeration basin in Period 4 are much lower than those obtained from the first
stage aeration basin in Periods 4 and 5.
Assays of the Period 4 samples were also conducted by SFRE. These analyses assayed for pathogenic and
potentially pathogenic bacteria (Salmonella, Shigella, Proteus, Pseudomonas, Mycobacterium tuberculosis,
fecal Streptococci, etc.) and enteroviruses, in addition to the microbiological indicators. All of these Period 4
aeration basin sample analyses for pathogenic bacteria and enteroviruses were negative. SFRE did quantify
the indicators, fecal coliform and the standard plate count. However, the SFRE levels were very low com-
pared to those found by the local laboratory, Aqualab.
The daily composite aeration basin samples taken in Period 5 were analyzed for selected pathogenic
bacteria and enteroviruses by UTSA and for the indicator organisms by Aqualab. Table 12 reports the
microbiological concentrations for these composite aeration basin samples. Comparison of the indicator
levels with those found in the grab samples from the first stage aeration basin show them to be quite com-
parable. The pathogenic bacteria selected for analysis were Proteus, Pseudomonas, fecal Streptococci,
Salmonella, and Shigella; the concentrations are reported as cfu/100 ml (colony forming units per 100
milliliters). Proteus, Salmonella, and Shigella were not detected in these samples. Pseudomonas and fecal
Streptococci, however, were consistently found, giving geometric means of 855 cfu/100 ml and 74.4 cfu/100
ml, respectively, Various other bacteria were also detected; these are reported in the footnote of Table 12.
Also noted is the loss of sensitivity for Proteus, Salmonella and Shigella in some samples due to overgrowth
of other organisms at high dilutions. In addition, the laboratory reported that a few of the samples were
received at a temperature above the desired 4°C.
74
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TABLE 9. UTSA MICROBIAL CHARACTERIZATION OF PERIOD 5
AERATION BASIN GRAB SAMPLES
Sample Temperature
Standard Analyses:
Proteus
Pseudomonas
Fecal Streptococci
Salmonella
Shigella
Coliphage
Enteroviruses- 3-day plaques
Enteroviruses-5-day plaques
Bacterial Screen:
Citrobacter
Clostridium
Edwardsiella
Enterobacter
EscherichiaH2S+
Fecal Coliform
Klebsiella
Leptospira
Mycpbacteria
Providencia
Serratia
Staphylococcus
Total Coliform
Total Plate Count
Yersenia
Distribution from Enterobacteriaceae
Oxidase positive
Enterobacter
Klebsiella
Proteus
Providencia
Escherichia
No growth
Not identified
9/28/76
6°C
cfu/100 ml
<30,000
3,300,000
720,000
<30,000
<30,000
pfu/fi
350,000
242
162
cfu/100 ml
<300
1,500
<300
20,000
<300
100,000
100,000
2,400
130,000
<300
<300
200,000
3,700,000
820,000,000
ND
Testing: %
68.4
7.0
1.7
-
-
-
22.8
—
*Solid fraction sample was contaminated; concentration
fraction sample (cf. Table 13).
10/11/76
9°C
cfu/100 ml
<50,000
970,000
1,000,000
<50,000
<50,000
pfu/B
230,000
*
*
cfu/100 ml
50,000
280
50,000
3,000,000
1,000,000
10,000,000
6,000,000
4,600
70,000
1,000,000
<50,000
300,000
1 10,000,000
580,000,000
ND
%
44.9
14.4
14.4
1.4
1.4
1.4
4.3
17.3
is greater than liquid
75
-------
TABLE 10. MICROBIOLOGICAL CONCENTRATIONS IN PERIOD 4 AERATION BASIN
SAMPLES TAKEN WITH HIGH VOLUME AEROSOL SAMPLING RUNS
High Volume
Run Number
Date
Standard
Plate Count
(No./lOO ml) X 10'
Total Coliform
(mfc/100 ml) X 106
Fecal Coliform
(mfc/100 ml) X 10"
Second Stage Aeration Basin Composite Sample
1
2
3.
4
5
6
7
2-9-76
2-10-76
2-11-76
2-12-76
2-13-76
2-14-76
2-15-76
Geometric Mean
Arithmetic Mean
Standard Deviation
600
80
33
28
72
86
78
80
140
204
0.22
0.42
0.56
0.22
0.09
0.46
0.52
0.31
0.36
0.18
0.002
0.012
0.0009
0.010
0.017
0.350
0.040
0.013
0.062
0.128
First Stage Aeration Basin Grab Sample
4
2-12-76
9,700
23
1.0
TABLE 11. MICROBIOLOGICAL CONCENTRATIONS IN PERIOD 5 FIRST STAGE AERATION BASIN
GRAB SAMPLES TAKEN WITH HIGH VOLUME AEROSOL SAMPLING RUNS
High Volume
Run Number
1
2
3
4
5
6
7
8
9
10
Geometric M
Date
10-1-76
10-4-76
10-4-76
10-5-76
10-6-76
10-7-76
10-7-76
10-7-76
10-8-76
10-8-76
ean
Arithmetic Mean
Standard Deviation
Standard
Plate Count
(No./ 100 ml) X 10'
640
1,270
146
2,870
1,260
490
360
700
580
220
607
854
805
Total Coliform
(mfc/100ml)X 10*
42
44
42
39
72
46
41
43
27
23
40.2
41.9
13.0
Fecal Coliform
(mfc/100 ml) X 106
2.9
2.1
1.7
1.9
6.1
2.2
2.5
1.2
2.0
0.76
2.03
2.34
1.45
Coliphage
(pfu/S) X 103
93
380
270
120
300
230
200
34
41
51
127
172
122
76
-------
TABLE 12. MICROBIOLOGICAL CONCENTRATIONS IN PERIOD 5 COMPOSITE
AND CHARACTERIZATION AERATION BASIN SAMPLES
Standard
Plate Count
Sample (No./lOOml)
Date
X 106
Total
Coliform
Fecal
Coliform
(mfc/100 ml) (mfc/100 ml)
X 106
X 106
Coliphage
(pfu/E)
X 103
Proteus
(cfu/lOQml)
X 103
Pseudomonas
(cfu/ 100 ml)
X 103
Bacteria
Fecal
Streptococci
(cfu/100 ml)
X 103
Enteroviruses
Salmonella
(cfu/ 100 ml)
X 103
Shigella
(cfu/ 100 ml)
X 103
3-Day
Plaques
pfu/B
5-Day
Plaques
pfu/B
Composite Samples
10-2-76
10-4-76
10-5-76
10-6-76
10-7-76
10-8-76
10-9-76
Geometric Mean
Arithmetic Mean
Standard Deviation
405
860
2,960
490
370
540
210
577
834
959
57
45
38
44
39
50
21
40.4
42
11.3
3.3
14.3
2.4
1.7
2.6
1.4
0.84
2.49
3.79
4.71
200
190
150
120
200
51
36
114
135
69.3
<0.33
<0.5
<1.0
<300*
<200*
<2,000*
<2,000*
1,400
1,700
440
480
800
180
4,600
855
1,370
1,520
86
20
2.1
590
330
15
1,200
74.4
320
445
<0.33
<0.5
<1.0
<300*
<200*
<2,000*
<2,000*
Characterization Samples
9-28-76
10-11-76
Note: Other Organisms
cfu/ 100 ml; 10-6
87,000
360
69
32
10-2 Escherichia-2.3 X 10
Serratia-8.0 X 106 pfu/100
10-8 Enterobacter-9.0 X 10'
•Sensitivity for Proteus,
cfu/ 100 ml.
Salmonella and Shigella lost
0.4
1.6
4 cfu/ 100 ml
350
230
<30
<50
3,300
970
720
1,000
, Enterobacter- 3.6 X 106 cfu/100 ml; 10-5 Serratia-3.0
ml, Citobacter, Enterobacter; 10-7 Enterobacter-3.1 X
due to overgrowth of other organisms at
fSolid fraction sample was contaminated; concentration is greater
than liquid
fraction sample
high dilutions.
(cf. Table 13).
<30
<50
<0.33
<0.5
<1.0
<300*
<200*
<2,000*
<2,000*
<30
<50
<24
192
503
284
134
't
111
114.0
203.3
171.5
242
t
X 106 pfu/100 ml, Enterobacter- 1.4 X
<19
192
503
284
146
t
277
114.0
204.9
171.0
242
t
10T
10' cfu/lOOml.Escherichia-l.OX 10» cfu/100 ml;
-------
The enterovirus concentrations are summarized in Table 12 and reported by sample fraction in Table 13.
The concentrations are given in pfu/J? (plaque-forming units per liter). For two of the samples (one composite
and one characterization), the solid fraction sample was contaminated. The values for these two samples are
reported as > (greater than), because the concentrations were greater than the upper detection level in the li-
quid fraction. The geometric mean concentrations of the 3-day and 5-day plaques were both 114 pfu/J,
indicating that most of the enteroviruses detected were 3-day plaques - presumably polioviruses (cf. Section
5). Comparison of the geometric mean concentrations of the solids and liquid fractions for the 3-day plaques
(202 pfu/Jt vs. 40 pfu/,0 indicates that the viable enteroviruses were primarily in the solids fraction.
TABLE 13. ENTEROVIRUS CONCENTRATIONS IN PERIOD 5 COMPOSITE AND
CHARACTERIZATION AERATION BASIN SAMPLES
Enterovirus Concentrations
Composite Samples
10/2/76
10/4/76
10/5/76
10/6/76
10/7/76
10/8/76
10/9/76
Characterization
Samples
9/28/76
10/11/76
3-Day Plaques
Total Count (pfu/2)
Solids Fraction (pfu/e)
Liquid Fraction (pfu/fi)
Concentration Efficiency (%)
<24
<23
< 0.04
50
192
190
1.5
48
503
502
0.71
98
284
279
5.0
30
134
65
69
30
>41
CS
41
89
277
276
0.94
78
242
202
40
92
>7.0
CS
7.0
25
5-Day Plaques
Total Count (pfu/6)
Solids Fraction (pfu/e)
Liquid Fraction (pfu/6)
Concentration Efficiency (%)
<19
<19
< 0.6
50
192
190
1.5
48
503
502
0.71
98
284
279
5.0
30
' 146
65
81
30
>43
CS
43
89
277
276
0.94
78
242
202
40
92
>7.0
CS
7.0
25
Inspection of the microbiological concentrations reveals that the distribution of some of the parameters
is highly skewed, due to one or more unusually high values. This becomes particularly evident when the
geometric and arithmetic means are compared; for the standard plate count, fecal coliform, Pseudomonas,
fecal Streptococci, and the 3-day and 5-day enteroviruses, these two means are quite discrepant.
The trace metal concentrations found in the aeration basin samples for Periods 4 and 5 are shown in
Tables 14 and 15. Whereas the composite samples for Period 4 were taken from the second stage aeration
TABLE 14. TRACE METALS CONCENTRATIONS IN PERIOD 4
SECOND STAGE AERATION BASIN COMPOSITE SAMPLES
Sampling
Date
2-9-76
2-10-76
2-11-76
2-12-76
2-13-76
2-14-76
2-15-76
Geometric Mean
Arithmetic Mean
Standard Deviation
Detection Limit
Trace Metal Concentration, mg/8
Cadmium
<0.0002
0.0003
0.0003
0.0005
0.0002
0.0003
<0.0002
0.0002
0.0003
0.0001
0.0002
Lead
<0.0025
<0.0025
< 0.00 25
<0.0025
<0.0025
<0.0025
<0.0025
0.0025
Mercury
0.000025
0.000071
0.000093
0.000061
0.000065
0.000036
0.000025
0.000039
0.000050
0.000031
0.000025
78
-------
TABLE 15. TRACE METALS CONCENTRATIONS IN PERIOD 5
FIRST STAGE AERATION BASIN COMPOSITE SAMPLES
Sampling
Date
10-2-76
10-4-76
10-5-76
10-6-76
1 0-7-76
10-8-76
10-9-76
Geometric Mean
Arithmetic Mean
Standard Deviation
Detection Limit
Trace Metal Concentration, mg/e
Cadmium
0.43
0.58
0.57
0.57
0.59
0.59
0.53
0.55
0.55
0.06
0.0002
Lead
1.00
1.31
1.26
1.47
1.47
1.35
1.18
1.28
1.29
0.17
0.0025
Mercury
0.0045
0.0073
0.0077
0.0053
0.0064
0.0051
0.0056
0.0059
0.0060
0.0012
0.000025
basin, the Period 5 samples were obtained from the first stage aeration basin. Inspection of the metal concen-
trations for the two periods reveals that the values are consistently much lower in the second stage aeration
basin. The Period 4 (second stage aeration basin) geometric mean concentrations of cadmium and mercury
are 0.0002 mg/1 and 0.000039 mg/1, respectively; none of the samples contained lead concentrations above
the lower detection level. By comparison, the Period 5 (first stage aeration basin) geometric mean concentra-
tions for cadmium, mercury and lead are 0.55 mg/1, 0.0059 mg/1, and 1.28 mg/1. Little daily variation was
observed in the metal concentrations, as evidenced by comparing the standard deviation to the arithmetic
mean to get a low coefficient of variation for each metal.
Air Samples
Meteorological and Sampling Conditions-
The meteorological conditions for all air sampling runs conducted during periods 1,2,4 and 5 are sum-
marized in Tables 16-18, along with the sampling times and the alignment of the sampler line. The data
presented were taken, whenever possible, from our aerosol run reports and meteorological instrumentation at
the sampling site, in the earlier sampling periods. When field instrumentation was lacking, the O'Hare Inter-
national Airport local climatological data were utilized. There were some differences between the O'Hare
meteorological data and that collected in the field, resulting largely from differences in the location and
height of the instrumentation. Whereas the field measurements were taken within a few hundred meters of the
aeration basin at a height of 3.5 m, the O.Hare measurements were located eight miles east-southeast of the
sewage treatment plant, with the wind instruments elevated 20 feet above ground and the temperature and
humidity at the 40 foot elevation.
During Period 4, the high volume and Andersen microbiological air samples for a given run were not
taken simultaneously, as they were in Period 5. Therefore, the sampling time reported is the total time for the
four or five successive samples taken during each run, with each individual sample taking thirty minutes. This
successive sampling procedure necessitated reporting the range of meteorological conditions over the Period 4
runs, as the wind direction, temperature, relative humidity, and sky cover usually did not remain constant
over the period during which the samples were taken.
79
-------
TABLE 16. METEOROLOGICAL SUMMARY FOR PERIOD 1 AND 2 HIGH
VOLUME PARTICULATE AND ANDERSEN AIR SAMPLES
Run
No.
High
Volume Paniculate Air Samples
Alignment of
Duration Sampler Line
of (Upwind Temperature
Start Start Sampling Direction*) Range
Date Time (hr) (deg) (°C)
Relative
Humidity
Range
Wind
Velocity
Range
(m/see)
Sky
Cover Start
(tenths) Date
Average
Start
Time
Duration
of
Sampling
(min)
Andersen Air Samples
Alignment of
Sampler Line Mean
(Upwind Relative Wind
Direction*) Temperature Humidity Velocity
(deg) (°C) (%) (m/sec)
Sky
Cover
(tenths)
Period I
1
2
3
4
5
6
7
10-13-74 0800
10-14-74 0800
10-15-74 0800
10-16-74 0800
10-17-74 0740
10-18-74 0745
10-19-74 0830
24
24
24
24.5
24.5
25
24.5
105
295
305
215
230
35
275
12.1 - 17.2
2.8-12.7
3.3-13.9
8.9-17.8
6.7 - 18.4
2.2 -8.3
2.8 - 10.0
84-90
55-86
44-86
38-66
33-83
68-89
57-85
1.5-5.5
1.8-5.5
0.9-3.1
1.5-4.9
3.1-5.5
1.5-3.7
1.5-4.6
10
0-10
1-7
0-1
0-10
7-10
10
10-13-74
10-14-74
10-15-74
10-16-74
10-17-74
10-18-74
10-19-74
0845
0835
0840
0845
0810
0815
0855
15
15
15
15
15
15
15
105
295
305
215
230
35
275
12.1
12.1
7.8
11.7
14.4
7.8
5.6
90
86
75
64
47
74
83
2.4
4.0
2.4
3.4
5.2
2.1
2.1
10
10
1
1
0
10
10
ft/rod 2
1
2
3
4
5
6
7
*0°
-26-75 1020
-27-75 0920
-28-75 0935 -
-29-75 0925
-30-75 0920
-31-75 0910
2-1-75 1005
, 360° = N
90° =E
180° = S
270° = W
23
24
25
24.5
24
25
23.5
280
125
80
280
5
30
45
-5 .6- -0.6
-1.7-2.2
2.2-3.9
-3.9 - 2.2
-5.6-0
-5.6-1.1
'-5.0- 1.7
64-85
62-85
65-96
69-85
72-85
64-79
62-92
0.9-5.5
0-4.3
1.2-5.5
0-6.7
1.5-3.7
1.8-3.7
0-3.4
4-10
10
9-10
0-10
10
8-10
0-10
1-26-75
1-27-75
1 -28-75
1 -29-75
1-30-75
1-31-75
2-1-75
1010
1005
1020
1010
itioo
0940
1040
15
15
15
15
15
15
15
280
125
80
280
5
30
45
. -5.0
-1.1
2.8
1.7
-1.7
-4.4
1.1
69
74
71
80
79
73
64
4.9
1.8
3.7
5.8
2.1
1.8
3.4
7
10
9
10
10
10
10
-------
TABLE 1 7. METEOROLOGICAL SUMMARY FOR PERIOD 4
AND 5 HIGH VOLUME PARTICULATE SAMPLES
Run
No.
High-Volume Particulate Aii Samples
Start
Date
Avg
Start
Time
Average
Duration
of
Sampling
(hr)
Alignment of
Sampler Line
(Upwind
Direction*)
(deg)
Temperature
Range
(°C)
Relative
Humidity
Range
(%)
Wind
Velocity
Range
(m/sec)
Sky
Cover
(tenths)
Period 4
1
2
3
4
5
6
7
7
2-9-76
2-10-76
2-11-76
2-12-76
2-13-76
2-14-76
2-15-76
2-16-76
1200
1500
1535
1330
1330
1230
1630
0730
23
24.5
20
22.5
22
25
18
21.5
160-215
230-320
170-270
0-180
120-340
120-230
30-255
30-120
3.3-11.1
1.7-12.1
1.1-12.7
6.1-12.7
-3.3-4.4
5.0-17.2
2.8-15.0
2.2-6.1
68-77
38-79
38-62
33-80
35-78
30-80
54-82
74-89
3.1-6.1
1.5-6.1
1.5-6.1
0.94.6
0.8-4.6
3.1-7.6
0.94.6
1.5-3.7
3-10
0-10
0-10
0-10
0-10
7-10
1-10
10
ft?nbd 5
1
2
3
4
5
6
7
9-30-76
10-1-76
10-2-76
10-3-76
104-76
10-6-76
10-7-76
1050
1200
1050
1000
1045
0945
1035
23.5
23
23.5
23
21.5
25
23.5
220-280
155-300
140-305
100-170
150-270
300-350
300-45
8.3-24.4
9.4-26.2
12.1-25.0
12.1-26.2
10.6-26.2
7.3-13.3
7.8-9.5
33-80
33-83
47-90
38-69
38-90
42-66
66-71
0-3.1
0-2.7
1.2-3.7
1.8-3.4
04.9
2.4-5.2
1.2-2.4
0-1
0
0-10
1-10
7-10
2-10
8-10
* 0° , 360° = N
90° =E
180° =S
270° =W
81
-------
TABLE 18. METEOROLOGICAL SUMMARY FOR PERIOD 4 AND 5 HIGH VOLUME
AND ANDERSEN SAMPLING RUNS
Run
No.
Date
Time
Alignment of
Sampler Line
(Upwind
Direction*)
(deg)
Wind
Direction
about
Sampler Line
(deg)
Temperature
(°C)
Relative
Humidity
(%)
Mean
Wind
Velocity
(m/sec)
Sky
Cover
(tenths)
Mean
Solar
Radiation
(g cal/cmj/min)
Period 4 (High Volume and Andersen Samplers)
1
2
3
4
5
6
7
2-9-76
2-10-76
2-11-76
2-12-76
2-13-76
2-14-76
2-16-76
1120-1730
1115-1645
0906-1543
1020-1900
0835-1458
0815-1452
0829-0859
0
220-295
320
210
3-12
120-180
45
2.2-6.1
7.8-11,1
2.2-4.4
7.3-10.6
6.1-3.3
0.6-7.8
3
71-79
72-74
36-70
55-72
39-60
30-76
71
4.2
3.9
4.1
4.7
3.9
3.5
1.8
10
10
0-6
2-10
10
2-8
10
Period 5 (High Volume Samplers)
1
2
3
4
5
6
7
8
9
10
10-1-76
10-3-76
10-4-76
104-76
10-6-76
10-6-76
10-7-76
10-7-76
10-8-76
10-8-76
1150-1220
1850-1920
0600-0630
1830-1900
1230-1300
1920-1950
0530-0600
1400-1430
0555-0625
1515-1545
195
90
125
135
0
0
325
0
295
265
(-40, +35)
(-15,0)
(-15, +40)
(-5, +25)
(-50, +5)
(-5, +15)
(-5, +45)
(-40, +5)
(+5, +45)
(-40, +25)
24.2
19.2
11.1
23.3
11.6
7.5
7.5
8.3
6.2
12.4
43
67
78
45
55
58
70
70
80
48
2.4
0.8
0.3
1.5
3.1
2.1
1.8
1.5
1.7
1.5
0
10
1
3
5
8
10
10
9
8
1.80
0.00
0.00
0.00
1.00
0.00
0.00
0.10
0.00
0.45
Period 5 (Andersen Samplers)
1
2
9-29-76
10-7-76
1215-1300
1445-1515
250
0
(-30, +30)
(-25, +15)
18.3
8.3
62
70
1.5-3.1
1.5
8-9
10
0.60
0.05
*0°,360° = N
90° = E
180° = S
270° = W .
The daily resultant wind directions at O'Hare International Airport were tabulated for the study period
during which the Egan plant was in operation (January through October, 1976). The wind rose diagram giv-
ing the distribution of daily resultant directions from which the wind blew over this operational period is
presented in Figure 12. The wind rose diagram indicates that the prevailing wind directions in the vicinity of
the Egan plant were primarily from the southwest and south, and secondarily from the north-northeast dur-
ing the operational period. Thus it is anticipated that there would be greater exposure to the microorganisms
in the aerosol emanating from the Egan plant's aeration basins for the residences located to the northeast,
south-southwest, and north than for residences in other directions. Exposure should be least for residences
situated west and northwest of the Egan aeration basins.
82
-------
LOCATION:
PERIOD:
OBSERVATION:
NO. OF OBSERVATIONS:
SOURCE:
O'HARE INTERNATIONAL AIRPORT
CHICAGO, ILLINOIS
JANUARY - OCTOBER, 1976
DAILY RESULTANT WIND DIRECTION
305 DAYS
LOCAL CLIMATOLOGICAL DATA,
NATIONAL CLIMATIC CENTER
330
300
W270
240
210
N
0/360°
30°
60°
120"
150"
180°
S
Figure 12. Distribution of daily wind directions during the operational period (Jan - Oct, 1976).
83
-------
Microbiology-
Microbiological Concentration and Isolation Daw—The microbiological concentrations obtained from
the air samplers are reported in Tables 19-21. Distances are measured from the nearest edge of the first stage
aeration basin in the alongwind direction. Table 19 gives the analytical results for the Andersen air samples
taken in Periods 1, 2 and 4. The pathogenic and potentially-pathogenic bacteria sought were Mycobacterium
tuberculosis, Proteus, Pseudomonas, Salmonella, Shigella, Staphylococcus Aureus and S. epidermidis, and
Streptococcus (alpha, beta, and gamma). No pathogenic bacteria were isolated in Periods 1 or 2. In Period 4,
Pseudomonas were isolated in three of the downwind samples and Streptococcus-alpha was found in one
sample. Table 20 gives the indicator concentrations and pathogen isolations found for the Period 4 high
volume samplers. Table 20 shows one sample with a Pseudomonas isolate, but because the air sample was col-
lected 1570 m upwind, the sewage treatment plant is unlikely to have been the source. Excluding the sample
concentrations inferred to be affected by contamination, the levels for the three microbiological indicators
have consistently higher values for the aerosol samples taken close downwind from the aeration basin.
Table 21 reports the microbiological concentrations for the Period 5 high volume sampling runs. These
samples were subjected to more extensive and quantitative analyses, including four microbiological indicators
(standard plate count, total coliform, fecal coliform, and coliphage), selected pathogenic bacteria (Proteus,
Pseudomonas, fecal Streptococci, Salmonella, and Shigella), and 3-day and 5-day enteroviruses. Of the
bacteria selected, no Proteus, fecal Streptococci, Salmonella, or Shigella were detected in any air sample.
However, Pseudomonas was found in every run. Klebsiella was also isolated in several samples, even out to a
distance of 300 meters from the plant. A single enterovirus (poliovirus type III) was detected in the air
samples; this was on Run 6 in a sample taken 300 meters downwind. The indicator levels show a rapid
decrease in concentration with distance from the plant. Other organisms isolated and their quantified concen-
trations are reported in the footnote of Table 21.
It is our experience from air sampling both at the Egan plant and at Pleasanton(44^ (conducted with the
same samplers both before and after Period 5) that high volume aerosol samplers are essential to obtain quan-
titative microbiological aerosol concentrations, especially on pathogens. However, the high volume aerosol
samplers have a tendency to become and remain contaminated with some of the hardy microorganisms, such
as Pseudomonas, in parts that are unaffected by our thorough routine cleaning procedures. The primary
evidence of sampler contamination is usually an extremely high standard plate count, which persists in the
sampler over several consecutive runs. Since these analytical results are not available to the field crew until the
next day, the sampler contamination problem is difficult to detect and correct in the field. As indicated in
Table 21, Sampler 3 shows this evidence of contamination in Runs 1 through 4. Laboratory notations in-
dicating possible sample contamination of either field or laboratory origin are also reported in Table 21.
Because the large values associated with sampler contamination will dominate the statistical analysis,
general criteria have been developed for inferring sample contamination that has affected the analytical value,
based on combined experience in Pleasanton and at the Egan plant. The values inferred to be affected by
sampler contamination have been underlined in Tables 20 and 21. The ensuing statistical analyses have been
conducted both excluding and including these underlined values. The specific criteria for inferring possible
sample contamination in the Chicago data, signified by underlining the affected value, are given below.
Standard plate count values>30,000/m3 were judged to be contaminated. The basis for this criterion
value was the observation of a consistent pattern of high values for a particular sampler over several runs, or
indication of contamination by the laboratory. Total and fecal coliform values were considered contaminated
if they showed an unusual value in addition to some indication of sampler contamination by the laboratory or
the corresponding standard plate count value. Pseudomonas concentrations were underlined in the table if
they showed unusual values along with standard plate count indications of sampler contamination, or if they
showed very unusual values coupled with sampler contamination in the run immediately preceding or follow-
ing the one in question. For Pseudomonas, this procedure resulted in a criterion value of>3000 cfu/m3.
84
-------
TABLE 19. PATHOGEN ISOLATIONS FROM PERIOD I, 2 AND 4 ANDERSEN AIR SAMPLES
Period
Date
Approx.
Distance
(m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No. /nr1)
Period 2
Date
Approx.
Distance
(.m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No./m3)
Period 4
Date
Distance
(m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No./m3)
Residential Upwind (>300 in)
10-13-74
10-14-74
10-15-74
10-16-74
10-16-74
10-17-74
10-18-74
10-19-74
Geometrk
1600 U
5000 U
1600 U
1600U
5000 U
1600U
1600 U
1600 U
Mean
Arithmetic Mean
Standard Deviation
None
None
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
32
34
61
37
35
2
8
34
22
30
18
1-27-75
1-27-75
1-28-75
1-28-75
1-29-75
1600 U
500 U
1600U
500 U
500 U
None
None
None
None
None
Bacillus
Bacillus
Various
Various
Bacillus
8
72
2
12
5
9
20
29
2-9-76
2-10-76
2-11-76
2-12-76
2-13-76
2-14-76
1632 U
1570 U
580 U
928 U
660 U
1348 U
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
15
14
21
66
19
51
26
31
22
Sewage Treatmenl Plant (^300 ml
10-13-74
10-14-74
10-15-74
10-16-74
10-17-74
10-17-74
10-18-74
10-19-74
Geometrk
SOD
50 D
SOD
50 D
5D
SOD
SOD
50 D
; Mean
Arithmetic Mean
Standard Deviation
None
None
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
11
309
61
511
27
52
25
18
53
127
184
1-26-75
1-27-75
1-28-75
1-29-75
1-30-75
1-31-75
2-1-75
2-1-75
SOD
SOD
50 D
SOD
50 D
SOD
5 D
SOD
None
None
None
None
None
None
None
None
Bacillus
Bacillus
Various
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
471
22
15
19
36
14
6
11
24
74
161
2-9-76
2-9-76
2-10-76
2-10-76
2-11-76
2-11-76
2-12-76
2-12-76
2-13-76
2-14-76
252 D
1 D
107 D
7 D
170 D
18 D
114D
20 D
26 D
1 D
None
(2) Pseudo-
monas
(1) Pseudo-
mo nas
None
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Various
Various
Bacillus
Bacillus
205
>1200
80
>1200
39
166
438
392
314
242
269
428
426
(continued)
-------
TABLE 19. (continued)
Period 1
Date
Approx.
Distance
(m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No./m3)
Period 2
Date
Approx.
Distance
(m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No./m3)
Period 4
Date
Distance
(m)
Pathogens
Predominant
Organism
Standard
Plate Count
(No./m3)
Residential Downwind (>300 m)
10-13-74
10-13-74
10-14-74
10-14-74
10:15-74
10-15-74
10-16-74
10-17-74
10-18-74
10-18-74
10-19-74
10-19-74
Geometric
500 D
1600D
1600D
5000 D
1600D
1600D
1600D
1600 D
1600D
5000 D
1600D
5000 D
Mean
Arithmetic Mean
Standard Deviation
None
None
None
None
None
None
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
18
245
258
24
339
38
12
159
33
185
135
77
131
115
1-26-75
1-26-75
1-26-75
1-27-75
1-28-75
1-29-75
1-29-75
1-30-75
1-30-75
1-30-75
1-31-75
1-31-75
1-31-75
2-1-75
2-1-75
2-1-75
500 D
1600 D
5000 D
1600 D
1600D
500 D
1600D
500 D
1600D
5000 D
500 D
1600D
5000 D
1600 D
1600 D
5000 D
None
None
None
None
None
None
None
None
None
None
None
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Various
Bacillus
Bacillus
Bacillus
Bacillus
None
Bacillus
Bacillus
Bacillus
Bacillus
Diphtheroid
Micrococcus
Bacillus
21
4
7
240
1
27
26
27
<1
44
39
22
15
8
258
12
16
47
80
2-9-76
2-10-76
2-11-76
2-11-76
2-12-76
2-12-76
2-12-76
2-13-76
2-13-76
2-13-76
2-13-76
2-14-76
2-14-76
2-14-76
2-14-76
660 D
830 D
2230 D
800 D
1500 D
844 D
384 D
4228 D
1892 D
1268 D
480 D
820 D
615 D
388 D
340 D
None
None
None
None
None
None
Pseudo-
mo nas
None
None
Strepto-
coccus-alpht
None
None
None
None
None
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
Diphtheroid
Bacillus
None
Bacillus
Strepto-
coccus-alpha
Bacillus
Bacillus
Bacillus
Bacillus
Bacillus
13
59
16
12
58
51
134
<1
24
114
8
60
60
41
26
27
45
38
oo
o\
-------
TABLE 20. MICROBIOLOGICAL CONCENTRATIONS IN PERIOD 4
HIGH VOLUME AEROSOL SAMPLING RUNS
Time
1120-1150
1305-1410
1519-1553
1700-1730
Distance
(m)
Sampler
No.
Standard
Plate Count
(No./m3)
Total
Coliform
(mfc/m3 )
Fecal
Coliform
(mfc/m3 )
Pathogens Isolated
Run No. 1,2-9-76
660 D
1632 U
252 D
1 D
1115-1145
1245-1315
1450-1520
1615-1645
1570 U
830 D
107 D
7 D
1
1
30,000
13,000
6,200
120,000
<0.3
<0.3
1.5
200
<0.3
<0.3
<0.3
8.9
None
None
Streptococcus-alpha
Streptococcus-alpha
Run No. 2,2-10-76
1
1
1
1
890
1,200
1,200
35,000
0.3
<0.3
1.2
210
Run No. 3, 2-11-76
0906-0936
1128-1158
1230-1300
1419-1449
1513-1543
580 U
2230 D
800 D
170 D
18D
1
1
1
1
590
890
1,200
590
8,900
0.3
<0.3
<0.3
<0.3
13
Run No. 4,2-12-76
1020-1050
1124-1154
1430-1500
1533-1603
1735-1805
1830-1900
928 U
1500D
844 D
384 D
114D
20 D
1
1
1
1
1
1
83,000
53,000
41,000
3,200
8,900
77,000
<0.3
<0.3
<0.3
<0.3
3.0
140
<0.3
<0.3
<0.3
8.3
Pseudomonas
None
None
None
<0.3
<0.3
<0.3
<0.3
0.3
None
None
Streptococcus-alpha
Streptococcus-alpha
None
<0.3
<0.3
<0.3
<0.3
<0.3
1.2
None
None
None
None
None
None
Run No. 5,2-13-76
0835-0905
1005-1035
1107-1137
1210-1240
1316-1346
1428-1458
660 U
4228 D
1892 D
1268D
480 D
26 D
1
1,500
1,200
1,500
890
1,800
13,000
<0.3
<0.3
<0.3
<0.3
0.3
4.7
<0.3
<0.3
<0.3
<0.3
<0.3
0.3
None
None
None
None
None
None
Run No. 6, 2-14-76
0819-0849
0945-1015
1047-1117
1152-1222
1300-1330
1422-1452
1348U
820 D
615D
388 D
340 D
1 D
1
1
1
1
1
1
1,200
650
970
590
3,500
14,000
<0.3
<0.3
<0.3
<0.3
0.3
10
<0.3
<0.3
<0.3
<0.3
<0.3
1.8
None
None
None
None
None
Streptococcus-alpha
Run No. 7, 2-16-76
0829-0859
3620 U
1,500
<0.3
<0.3
None
Underlined value: sample contamination affecting result inferred.
Note: Other organisms isolated were Bacillus, E. coli. Enterobacter. Diphtheroid and Mictococcus.
87
-------
TABLE 21. MICROBIOLOGICAL CONCENTRATIONS IN PERIOD 5
HIGH VOLUME AEROSOL SAMPLING RUNS
Run No.
(Date) Distance
Time (meters)
1 500 U
(10-1-76) 15 D
1150-1220 250 D
310 D
1000 D
1600 D
2 960 U
(10-4-76) 25 D
1850-1920 50 D
100 D
200 D
500 D
3 960 U
(10-4-76) 25 D
0600-0630 50 D
100 D
225 D
1000 D
Sampler
Number
6
5
7
2
4
3*
8
3*
2
5
7
4
8
3*
2
5
7
4
* Contaminated Sampler.
Underlined value: sample contamin.
Indicators
Standard
Plate Count
No. /m3
120
510.000
950
530
500
200*
270*
2. 900,000
28,000
980
23 . OOP
2.900
400
1.900.000
1.300
2.000
310
1,200
ition.
Uion affecting
Total
Coliform
mfc/m
0.6
1.3
10.3
10. 3
10.3
Si'
2.7
93
2. 7
0.3
2.6
10.3
2.3
47
23
3.7
0.5
10.3
result inferred
Fecal
Coliform
mfc/m3
1 0.3
0. 5
10.3
1 0.3
1 0.3
*lf
0.7 '
I. 3
1 0.3
10.3
0.3
1 0.3
1 0.3
1 0.3
2.7
0.8
1 0.3
1 0.3
Coliphace
pfu/m
iO. 15
1 0. 13*
1 0. 13
< 0. 17*
1 0. 17*
10.17*
10.17
10.17
0.17
10.13
10.13
1 0. 14
1 0. 17
1 0. 17
1 0. 17
1 0. 13
1 0. 13
1 0. 14
Proteus
cfu/m3
1 10
1 9
1 9
1 II
1 9
1 II
1 II
1 II
1 II
1 9
1 9
< 9
1 17
1 II
1 II
1 9
i 13
i 9
Patho
Pseudomonas
cfu/m3
1 10
<9
34
22
36
8,700*
i 11
1 II
I, 100
1 9
26
27
I.IOO
33,000
2,500
1 9
1 13
< 9
;enic Bacte
Fecal
Streptococci
cfu/m3
< 10
1 9
1 9
1 II
1 9
1 II
1 II
1 II
< U
< 9
1 9
1 9
< 17
< 11
1 II
1 9
1 13
1 9
r i a
Salmonella
cfu/m3
1 10
< 9
< 9
1 II
i9
1 II
1 II
ill.
1 II
1 9
1 9
1 9
1 17
1 II
1 II
1 9
< 13
1 9
Shigella
cfu/m3
1 IO
1 9
i 9
i 11
i 9
1 U
1 II
< II
1 II
1 9
1 9
i 9
1 17
1 II
1 II
1 9
1 13
i 9
3 -Day
Plaques
pfu/m
10. 18
10.16
<0. 18
1 0.23
1 0. 16
< 0.20
< 0. 20
« 0.20
1 0. 20
< 0. 16
i 0. 15
^ i 0. 16
< 0.20
< 0. 20
1 0.20
< 0. 16
< 0. IS
1 0. 16
5 -Day
Plaques
pfu/m3
i 0.38
1 0.35.
" 0.36
1 0. 50
1 0.35
1 0.43
1 0.43
i 0.43
1 0.43
< 0.35
< 0. 33
1 0.35
1 0.43
1 0.43
1 0.43
1 0.35
1 0.33
< 0.35
Klebsiella
Isolated
No
Yes
No
No
No
No
No
No
No
No
; No
No
No
Possible
No
Yes
No .
No
00
00
(continued)
-------
TABLE 21. (continued)
Indicators
Run No. Standard
(Date) Distance Sampler Plate Count
Time (Meters) Number No. /m3
4 960 U 8 5,000
(10-5-76) 20 D 3* 4. 500,000
1830-1900 SOD 2 1 1 0, OOP
50 D 7 5.900
100 D 5 3.000
300D 4 1.400
5 660 U 1 32,000
(10-6-76) 15 D 8 8,000
1230-1300 95 D** 2 8,800
125 D*» 5 2,000
175 D** 7 9.700
300 D** 4 ' 6, 100
6 660 U 1 65,000
(10-7-76) IS D 8 120,000
1920-1950 95 D** 2 130.000
125 D** 5 11.000
175 D** 7 620
300 D** 4 4,600
Total Fecal
Coliform Coliform
mfc/m mfc/m
17
5jiOp_
15
38
12
2.2
0.3
1.0
7.3
2.4
0.8
0.3
<0.3
2.0
4.7
2.4
1.8
0. 5
* Contaminated Sampler.
* Laboratory Indication of contamination.
** Runs 5, 6, 7 and 8 - The second stage aeration basin occupied
Underlined value: sample contamination affecting result inferred
0.3
2100
< 0.3
0. 5
< 0. 3
< 0.3
< 0.3
< 0.3
4.7
< 0.3
< 0.3
< 0.3
< 0.3
0.3
0.3
< 0. 3
< 0.3
< 0.3
most of the
Coliphace
pfu/m
<0.17f
< 0. 17*
< 0. 17*
< 0. 13*
< 0. 13
< 0. 14
< 0. 15
0.17
< 0. 17
< 0. 13
< 0. 13
< 0. 14
< 0.15
< 0.17
< 0. 17
< 0. 13
< 0. 13
0.27
Pathogenic Bact
Proteus
cfu/m3
< 17
< 11
< 11
< 13
< 9
< 9
< 15
< 17
< 17
< 13
< 13
< 14
< 15
< 17
< 17
•s 13
•: 13
< 14
Fecal
Pseudomonas Streptococci
cfu/m cfu/m
600
470.000
< 11
< 13
< 9
9
< 15
< 17
U, OOP
< 13
13
< 14
< 15
< 17
< 11
< 13
< 13
14
space between the 15/20m downwind
< 17
< 11
< 11
130
< 9
< 9
< 15
< 17
< 17
< 13
< 13
< 14
< 15
< 17
< 17
< 13
< 13
< 14
sampler and the
eria
Salmonella
cfu/m3
< 17
< 11
< 11
< 13
<9
<9
< 15
< 17
< 17
< 13
< 13
< 14
< 15
< 17
< 17-
< 13
< 13
< 14
next downwind a
Shigella
cfu/m3
< 17
< 11
< 11
< 13
< 9
< 9
i 15
t 17
* 17
t 13
< 13
* 14
< 15
< 17
< 17
•: 13
< 13
•c 14
ampler.
3 -Day
Plaques
pfu/m3
<0. 20
<0. 20
< 0.20
<0. 15
<0. 16
<0. 16
< 0.24
< 0.23
< 0.23
< 0. 19f
< 0. 18
<'0.38
< 0.27
< 0.30
< 0. 23
< 0.21
< 0.21
0.22
5 -Day
Plaques
pfu/m
< 0.43
< 0.43
< 0.43
< 0.33
< 0.35
< 0. 35
< 0.47
< 0.47
< 0.47
< 0.37
< 0.36
< 0.76
< 0. 53
< 0.60
< 0.47
< 0.43
< 0.41
< 0.43
Klebsiella
Isolated
No
No
No
No
No
Yes
No
No
No
No
Yes
No
No
No
No
No
No
No
(continued)
-------
TABLE 21. (continued)
Run No.
(Date)
Time
7
(10-7-76)
0530-0600
8
(10-7-76)
1400-1430
9
(10-8-76)
0555-0625
10
(10-8-76)
1515-1545
Distance
(Meters)
820 U
20 D
75 D»*
125 D»*
200 D**
2000 D«*
660U
15 D
97 D**
125 D*»
175 D**
420 D»»
820 U
20 D
50 D
80 D
115 D
300 D
500 U
22 D
22 D
SOD
600 D
1600 D
Sampler
Number
4
8
1
2
5
7
4
8
2
5
7
1
4
8
2
5
7
1
4
5
2
7
1
8
Indicators
Standard
Plate Count
No. /m3
8,700
670, 000
8,800
21, 000
13,'DOO
330
19,000
2,800,000
1, 100,000
7,600
8. ZOO
68, OOP
130,000
43,000
1,700
28,000
800
6,300
19,000
29.000
21.000
130
5.000
170
Total
Coliform
mfc/m
<0.3
4.7
2. 3
1.0
1. 0
0.3
0.8
5.0
7.0
2.7
3. 1
3.0
<0.3
6.0
2.7
0.3
0.3
0.9
1.4
2.0
33
1.3
0.6
0.7
NOTE: Other organisms isolated (and quantified concentrations):
Providencia, and Serratia - (100/m3 -TNTC).
* Contaminated Sampler.
M Muslim
Data.
P
8
Fecal
Coliform Coliphage
mfc/m3 pfu/m3
<0.3 0.14
0.3 < 0. 17
< 0.3 < 0. 15
< 0.3 < 0. 17
< 0.3 < P. 13
< 0.3 < 0. 13
< 0.3 < 0. 14
0.3 < 0. 17
0.7 < 0.17
<0.3 < 0. 13
<0.3 <0.13
< 0.3 < 0. 15
< 0.3 < 0. 14
0.3 0.17
0.7 < 0. 17
< 0. 3 < 0. 13
< 0.3 < 0.13
< 0.3 < 0. 15
<0. 3 < 0. 14
<0. 3 0.27
<0.3
-------
Microbiological Aerosol Particle Size Distribution-Two Andersen sampler runs were conducted during
Period 5 in which microbiological levels were determined for each of the six sampler stages corresponding in
six particle sizes. For Run 1 (September 29, 1976), standard plate count levels were reported; for Run 2
(October 7, 1976), total coliform levels were determined. Table 22 reports the concentrations and percentage
distributions for the standard plate count run and/or the total coliform run according to sampler distance and
the particle diameter range for each of the six stages. In determining the percentage distributions, TNTC (too
numerous to count), CS (confluent), and M (missing plate) values were ignored, because no reasonable
numerical value could be found for substitution.
Examination of these tables suggests that the majority of the viable particles in the aerosol samples were
in the respirable range, with most of them falling between 3.3 and 7.0 microns in diameter. As would be
expected, the standard plate count data showed a broader range than the total coliform, including a sizable
number of particles greater than 7.0 microns in diameter. There seemed to be no clear trend for the particle
size distributions to change with sampler distance.
Microbiological Source and Distance Ana/ysis-The statistical treatment of the microbiological aerosol
data attempted to provide answers to two research questions through the use of paired comparisons. The first
of these, whether the Egan sewage treatment plant was implicated as a source of the microbiological
parameters measured, was considered through comparisons of upwind air samples, used as a measure of
background aerosol levels, and the closest downwind air samples. For parameters showing significantly
higher levels in the close downwind samples, thereby implicating the plant as a source, a further question was
considered: are the downwind levels of these parameters a health concern? In considering this question, the
more distance, residential downwind air samples were compared against the corresponding upwind air sample
for each run.
A general methodological approach for conducting the necessary paired comparisons was formulated.
The preferable method for such comparisons is the paired sample t-test which provides a very powerful test of
the null hypothesis that the average difference between pair members is equal to zero. This test, however, re-
quires two assumptions: the distribution of difference scores must be normal in shape and must be composed
of a continuous variable. The difference scores for the microbiological aerosol data were generally not
normally distributed due to the occurrence of isolated high values. With this type of microbiological aerosol
data, comparisons utilizing proportions are considered more relevant than comparisons of differences. Since
the sample sizes were typically quite small (less than 20), traditional tests for normality (skewness, kurtosis,
goodness-of-fit) were not appropriate. Simple inspection of the data and its coefficient of variation served to
determine if the difference scores distribution was normal. When the data were not normal, an attempt to
normalize it through transformation was made. For example, a natural log transformation was useful when
the differences were proportional. In cases where no normalizing transformation could be found, the t-test
technique was not employed. In addition, for some cases, the assumption of variable continuity was violated;
this was true for data containing many concentrations close to or below the detection level.
In cases where the data were not amenable to a paired sample t-test, the Wilcoxon one sample signed
ranks test was chosen as a first alternative. This test does not require the assumptions of normality and
variable continuity, as it is based on simple order relationships among the difference scores. Even here it was
occasionally necessary to use natural log transformed data to account for the presence of proportional dif-
ferences in the data rather than absolute values. Following Brownlee's suggestion^ (p. 260), pairs leading to
difference scores of zero were deleted from the data.
A second alternative, the sign test, was chosen for use in cases when even the rank order statistic was
deemed inappropriate. This was true when the data seemed to be unreliable, as in the case of unusually large
values appearing in the same run as very small values. The sign test is much more appropriate for such data,
as it ignores the magnitude of a difference, and makes use simply of its direction.
91
-------
TABLE 22. AEROSOL PARTICLE SIZE DISTRIBUTION FOR
STANDARD PLATE COUNT AND TOTAL COLIFORM
Standard Plate Count Concentration* (No. /m3)
Sampler
Numbe r
1
6
2
3
4
5
Distance
(me ten)
Upwind
10 D
20 O
50 D
100 O
200 D
>7.0
(Stage 1)
166
111
168
TNTC
68
TNTC
4.7-7.0
(Stage 2)
65
71
100
26
31
14
Range of Particle
3.3-4.7
(Stage 3)
TNTC
113
ill
19
25
11
Sizes (n
2. 1-3
(Stage
11
68
36
14
13
14
microns)
3 1.1-2.1
4) (Stage 5)
12
36
25
21
13
4
0.65-1
(Stage
2
0
. 1
6)
TNTC
2
2
0
Standard Plate Count Percentage Distribution
Sampler
Number
1
6
2
3
4
5
Total CoLiform
Sampler
Number
3
2
1
6
4
5
Total Coliform
Sampler
Numbe r
3
2
12
6
4
5
Distance
(meters)
Upwind
10 D
20 D
50 D
100 D
200 D
>7. 0
(Stage 1)
657.
27%
38%
TNTC
45%
TNTC
Concentrations (mfc/m^)
Distance
(meters)
Upwind
15 D
97 D
125 D
175 D
420 D
>7.0
(Stage 1)
0
0
5
0
0
cs
Percentage Distribution
Distance
[meters)
Upwind
15 D
97 D
125 D
175 D
420 D
>7.0
(Stage 1)
0%
0%
33%
0%
0%
CS
4. 7-7.0
(Stage 2)
25%
18%
23%
31%
20%
33%
4.7-7.0
(Stage 2)
0
5
5
0
1
CS
4.7-7.0
(Stage 2)
0%
67%
33%
0%
50%
CS
Range of Particle
3.3-4.7
(Stage 3)
TNTC
29%
25%
23%
16%
25%
Range of Particle
3.3-4.7
(Stage 3)
1
2
2
0
1
0
Range of Particle
3.3-4.7
(Stage 3)
100%
31%
17%
0%
50%
0%
Sizes (microns!
2. 1-3.3
(Stage 4)
4%
17%
8%
17%
9%
33%
Sizes (microns)
2. 1-3.3
(Stage 4)
0
0
2
0
0
0
Sizes (microns)
2. 1-3.3
(Stage 4)
0%
0%
17%
0%
0%
0%
1. 1-2. 1
(Stage 5)
5%
9%
6%
26%
9%
8%
1. 1-2. 1
(Stage 5)
0
0
0
0
0
CS
1. 1-2. 1
(Stage 5)
0%
0%
0%
0%
0%
CS
0.65-1. 1
(Stage 6)
1%
0%
TNTC
3%
2%
0%
0.65-1. 1
(Stage 6)
0
0
0
0
0
M
0.65-1. 1
(Stage 6)
0%
0%
0%
0%
0%
M
TNTC - Too numerous to count.
CS - Contaminated sample.
M - Missing data.
92
-------
The data from Periods 4 and 5 were combined when possible to increase sample sizes and resulting
statistical power. In Period 4, samples were taken with both the Andersen and high volume samplers. These
data were not combined, as their values are not comparable.
Some runs had missing or contaminated data for some of the air samples taken or laboratory analyses
conducted. If the upwind concentration for a particular parameter was unusable for any run, that run was
deleted from the analysis, as it had no valid measure of background concentrations for that parameter. If the
closest downwind sample could not be used, the next closest one was substituted, with the restriction that no
sample taken at a distance at or beyond 300 meters be considered a close downwind sample. All samples taken
at a distance of at least 300 meters were designated as residential downwind samples. If, in the comparisons
between upwind microbiological levels and residential downwind levels, more than one air sample fell into the
residential category, the geometric mean was used as the comparison value.
As previously discussed, inferences of possible sample contamination were made in some cases in addi-
tion to contamination reported by the laboratory. The statistical analyses potentially affected by such con-
tamination (standard plate count, total and fecal coliform, and Pseudomonas) were conducted twice,
therefore; once including these suspect values, and once deleting them from the analysis. In no case did
removal of the inferred contaminated samples change the significance or nonsignificance of the test statistic
employed. The probability levels reported in the following paragraphs are those from the analyses excluding
the possibly contaminated values. All tests of the null hypothesis of no difference were evaluated with two-
sided levels of significance, in order to allow for possibly significant differences in an unpredicted direction.
Source and distance analyses were performed only on those microbiological parameters which were
detected in sufficient quantity to warrant statistical treatment. Four parameters met this criterion: standard
plate count, total coliform, fecal coliform and Pseudomonas. The results of these analyses conducted are
summarized in Table 23, and a more detailed discussion follows in the text.
The Period 4 and 5 high volume aerosol sampler data were combined, and comparisons were made of the
upwind and closest downwind samples. A paired sample t-test performed on natural log transformed
standard plate count data from the. high volume samplers revealed that the close downwind samples had
significantly higher counts (p = .025). A similar test performed on natural log transformed Andersen sampler
data from Period 4 also found significantly higher levels in the close downwind samples (p = .008). This im-
plicates the Egan sewage treatment plant as a source of bacterial aerosol as measured by the standard plate
count. However, further comparisons of natural log transformed upwind levels versus residential downwind
levels showed there to be no significant differences between them. This was true for both the high volume and
the Andersen sampler data.
TABLE 23. SUMMARY OF SOURCE AND DISTANCE ANALYSES PERFORMED ON
PERIOD 4 AND 5 AIR SAMPLE MICROBIOLOGICAL DATA
Microbiological Parameter
Standard Plate Count
High Volume Samplers
Andersen Samplers*
Total Coliform
Fecal Coliform
Pseudomonas
Local Source Analysis
Significant source, P = 0.025
Significant source, P = 0.008
Significant source, P< 0.001
Significant source, P = 0.01
Not significant source
Residential Distance Analysis
No effect at residential distances
No effect at residential distances
No effect at residential distances
No effect at residential distances
-
*Period 4 data only.
93
-------
Similar paired sample t-tests were performed on the natural log transformed period 4 and 5 total
coliform concentrations. The comparison of upwind versus closest downwind levels revealed a highly signifi-
cant t-value (p <.001), suggesting that the sewage treatment plant was a source of total coliform. However,
just as for the standard plate count data, no significant difference was found between upwind concentrations
and residential downwind levels.
Because a great number of fecal coliform concentrations were close to, or below, the detection level, a t-
test was inappropriate. Therefore, a Wilcoxon signed ranks test was performed on natural log transformed
Period 4 and 5 data. This test yielded a significant rank sum (p = .01) for the comparison of upwind level ver-
sus closest downwind level, implicating the plant as a source of this microbiological indicator. No significant
difference was found between upwind fecal coliform levels and residential downwind levels.
Of the various other microbiological parameters subjected to laboratory analyses in the Period 5 high
volume air samples, only Pseudomonas was detected in enough quantity to warrant statistical analysis. A
signed rank test of the natural log transformed concentrations for upwind samples versus closest downwind
samples showed that the close downwind levels were not significantly greater than the upwind levels. This sug-
gests that the sewage treatment plant is not a significant source of Pseudomonas.
Microbiological Sampling Variability-It was possible to obtain a measure of the variation in measured
microbiological aerosol concentrations between samplers from two high volume sampling runs in Period 5,
during which two air samples were taken simultaneously at the same distance with different samplers placed
side by side. A coefficient of variation was determined for both runs for each of four microbiological
parameters: standard plate count, total coliform, fecal coliform, and Pseudomonas. The two coefficients
were then averaged for each parameter; this procedure yielded an average variation coefficient of 0.76 for
standard plate count, 0.93 for total coliform, 0.38 for fecal coliform, and 0.58 for Pseudomonas. With such a
limited amount of data, these sampling variability estimates are subject to much uncertainty.
More precise measures of the between sampler variation for high volume air samplers have been obtained
from SwRI's environmental monitoring study of spray irrigation aerosols at Pleasanton, California(44). This
study has included eight special quality assurance runs in each of which eight or more high volume air
samplers (the same samplers and operators used in Period 5 at the Egan plant) were placed side by side at the
same distance from the line of sprayers. The between sampler coefficients of variation at Pleasanton have
averaged 0.45 for standard plate count, 0.35 for total coliform, 0.39 for fecal coliform, 0.64 for coliphage,
0.40 for Pseudomonas, and 0.61 for fecal Streptococci, excluding the values inferred to have been affected by
sample contamination.
A comparison was made of the sampler variability estimates given above with the variability in the paired
upwind vs. downwind comparisons of the source and distance analysis. Only the standard plate count and
total coliform parameter data were sufficiently quantitative to warrant this comparison. For both
papameters, the standard deviations of the paired comparisons of upwind with both close and residential
downwind distances were an order of magnitude higher than the between sampler standard deviations. These
large paired comparison deviations are probably due to large local differences in the background levels of
standard plate count and total coliform at the upwind and downwind locations that were sampled
simultaneously. Considering these large local differences and the relatively small number of sampling runs
conducted, it is very unlikely that small true differences between the upwind and downwind levels of any
sampled parameter (i.e., differences of the same order of magnitude as the upwind levels) that might be pro-
duced by the Egan plant could be statistically detected from the air sampling data. Therefore, negative find-
ings in the source and distance analysis (i.e., that the Egan plant is not a significant source or has no effect at
residential distances) indicate only that the Egan plant does not make the predominant contribution to the
downwind levels.
Model of Human Microbiological Aerosol Exposure-It has been our intent to develop a limited model of
likely human exposure to microbiological aerosols emanating from a sewage treatment plant, including con-
94
-------
sideration of source variation, diffusion factors, aerosolization factors, and factors causing microbiological
die-off both in the initial shock of aerosolization and subsequently with aerosol age. The microbiological
aerosol concentration data obtained on this study are inadequate to warrant mathematical model develop-
ment. However, our Pleasanton spray irrigation study*44) has provided a much more complete
microbiological aerosol concentration data base under a variety of meteorological conditions, from which a
valid microbiological aerosol exposure model may be developed.
Our Pleasanton results do indicate that long-term cumulative microbiological aerosol exposure detec-
table through clinical specimen microbiology and disease and symptom incidence should show a negative ex-
ponential decrease with distance from the source. Because of the cyclical wind direction pattern associated
with frontal passage in a continental climate, the direction of a residence from the Egan plant should have
much less effect on long-term exposure than does its distance. While short-term exposure depends strongly on
solar radiation and other meteorological factors, long-term cumulative exposure is less influenced by
meteorological factors because it integrates over the daily cycle in these factors. Therefore, the negative ex-
ponential of distance from the Egan plant can be considered as a prime surrogate measure of long-term
cumulative exposure to its microbiological aerosol.
Trace Metals-
Trace Metal Concentration Dara--The trace metal concentrations obtained from the particulate air samples
taken in Periods 1, 2, 4, and 5 are presented in Tables 24 and 25. Table 24 reports the concentrations from
Periods 1 and 2, before the sewage treatment plant was in operation. These samples were taken at sites ap-
proximating the locations of the operational period samples, to get baseline levels of the trace metals to be
measured. The baseline levels are fairly uniform across the various locations, with considerable variability
among samples.
Table 25 reports the trace metal concentrations for the Periods 4 and 5 particulate air samples. The con-
centrations have been categorized into three general sampling areas (residential upwind, sewage treatment
plant, and residential downwind) and the arithmetic and geometric means and the standard deviations
calculated for each group. The samples taken within a 300 meter radius of the first stage aeration basin do not
seem to have noticeably higher concentrations of cadmium or lead, suggesting that the plant makes a negligi-
ble contribution to the levels of these metals in the air. Figures 13 and 14 plot the cadmium and lead concen-
trations for each day's samples with distance for Periods 4 and 5. Each day's samples have been connected
with solid lines and the day has been enumerated at both ends of each daily line. The high upwind lead con-
centrations on days 6 and 7 of Period 5 were sampled just downwind of a 30-35,000 car per day highway.
These figures confirm that the cadmium and lead concentrations show no consistent pattern of increase or
decrease with distance.
The mercury concentrations do seem to have most of their highest values in the vicinity of the sewage
treatment plant. The operational period values are also generally higher in all three locations than those ob-
tained in the April 1975 sampling (Table 24). The Period 4 levels tend to be higher than those in Period 5. The
off-scale results, which exceeded the upper detection limit, have not been included in calculating the means
and standard deviations. These off-scale results could be due either to valid high values or to sample con-
tamination.
Figure 15 shows the mercury concentrations plotted against distance. The Period 4 data were plotted
twice, because of the presence of off-scale values in the samples collected at the aeration basin. In the upper
graph, the off-scale values are plotted at an arbitrary cut-off point; in the lower graph, they have been
deleted. The final graph shows the Period 5 mercury concentrations plotted against distance. These three
plots, when considered together, seem to provide some evidence that the mercury levels are elevated within the
plant boundaries.
Trace Metal Source and Distance Analysis-The statistical analysis of the trace metal concentrations found
in the particulate air sampling was similar to that of the microbiological concentrations. Similar questions
95
-------
TABLE 24. TRACE METAL CONCENTRATIONS IN PERIODS 1 AND 2
PARTICIPATE AIR SAMPLES
Location Date
Residential 10-13-74
Upwind 10-14- 4
(-300 ml 10-15- 4
10-16- 4
10-16- 4
10-17- 4
10-17- 4
10-18- 4
10-19-74
Geometric Mean
Arithmet c Mean
Standard Deviation
Sewage 0-13-74
Treatment 0-14-74
Plant 0-15-74
( <300 m) 0-16-74
0-17-74
10-18-74
10-19-74
Geometric Mean
Arithmetic Mean
Standard Deviation
Residential 10-13-74
Downwind 10-13-74
( >300 m] 10-14-74
10-14-74
10-15-74
10- 6- 4
10- 7- 4
10- 8- 4
10- 8- 4
10- 9- 4
10- 9- 4
Geometric Mean
Arithmetic Mean
Standard Deviation
Detection Limit:
* Sample tube broken
Distanc
(n-.eters
1600 U
5000 U
1600 U
5000 U
1600 U
5000 U
3600 U
1600 U
1600 U
50 D
50 D
50 D
50 D
50 D
50 D
50 D
500 D
1600 D
1600 D
5000 D
1600 D
1600 D
1600 D
1600 D
5000 D
1600 D
5000 D
Period 1
Cadmium
0.0024
0.0031
0.0014
0.0013
0.0013
0.0011
0. 0024
0. 0005
0. 0013
0.0015
0.0016
0.0008
0.0021
0. 0006
0.0018
0.0031
0.0010
0.0005
0.0015
0.0013
0.0015
0. 0009
0.0020
0. 0006
0. 0015
0.0014
0.0015
0. 001 1
0. 0005
0. 0026
0. 0015
0. 0027
0.0011
0. 0013
0.0015
0. 0007
0.0003
Copper
0. 100
0.099
0. 121
0.058
0.069
0. 108
0. 083
0. Oil
0. 186
0.076
0.093
0.048
0.086
0.077
0. 143
0. 076
0.082
0.046
0.068
0.078
0.082
0. 030
0. 092
0.844
, 0.175
1.261
0.143
0.041
0.074
0.084
0. 154
0. 132
0.252
0. 169
0.296
0.390
0. 0043
Lead Zinc
0.60 0.347
0.33 <0.025
0.95 0.185
0.46 0.185
0.29 0.185
0.38 0.031
0.32 0.127
<0. 04 0.197
0.30 0.150
0.30 0. 11
0.41 0.16
0.26 0.10
0.69 0.382
0.48 0.127
2.27 0.116
0.33 0.231
0.51 0.081
0.39 0.047
0.33 0.139
0.56 0.13
0. 71 0. 16
0.70 0.11
0.42 0.289
0.25 0.173
1.15 0.197
1.86 0.220
1.46 0.208
0.52 0.324
2.59 0.097
0.51 0.139
0.71 0.231
0.26 0.127
0.37 0.108
0.68 0.18
0.92 0.19
0. 76 0. 07
0.0448 0.0248
Date
1-27-75
1-27-75
1-28-75
1-29-75
-26-75
-27-75
-28-75
-29-75
-30-75
-30-75
-31-75
-31-75
2-1-75
2-1-75
-26-75
-26-75
-26-75
-27-75
-28-75
-29-75
-29-75
-30-75
-30-75
-31-75
-31-75
2-1-75
Distance
(metersl
1600 U
500 U
500 U
500 U
50 D
50 U
50 U
50 U
5 D
50 D
5 D
50 D
5 D
50 U
500 D
1600 D
5000 D
1600 D
1600 D
500 D
1600 D
500 D
1600 D
500 D
1600 D
5000 D
Period 2
Cadmium
0. 0028
0. 0023
0. 0028
0. 0043
0.0030
0. 0030
0. 0009
0. 0009
0. 0049
0.0029
0. 0007
0. 0040
0. 0010
0. 0020
0.0009
0.0016
0.0008
0. 0016
0. 0020
0.0015
0.0038
0.0004
0.0012
0.0034
0. 0055
0.0034
0. 0024
0.0035
0. 0015
0. 0009
0.0018
0. 0009
0. 0019
0. 0024
0.0015
0.0003
Copper Lead
0.132 1.01
0.041 0.74
0.047 0.97
0.263 0.45
0.090 0.76
0.121 0.79
0. 104 0. 26
0.099 0.77
0.186 2.13
0.164 1.03
0.132 0.69
0. 044 0. 38
0.143 0.41
0.019 0.63
0. 132 0. 91
0.012 0.59
0.107 1.34
0.078 0.78
0. 104 0. 89
0.060 0.52
0.020 0.83
0.056 0.32
0.329 0.67
0.384 1.56
0.406 0.93
0.033 0.42
0. 105 0. 37
0.329 0.29
0.026 0.32
0.274 0.61
0.020 0. 53
0.461 0.59
0.113 0.55
0. 203 0. 62
0.175 0.36
0. 0043 0. 0448
Mercurv
Zinc Date
0.382 -17-75
0.197 -23-75
0.312 -23-75
0.049 -23-75
-23-75
-24-75
-24-75
0. 18
0.24
0. 15
0.197 4-14-75
0.775 4-15-75
0.347 4-23-75
0.127 4-23-75
01 113
0. 104
0. 162
0. 220
0. 025
0. 106
0. 15
0.22
0.21
0.185 4-15-75
0.061 4-15-75
0.231 4-15-75
0. 532 4-24-75
0.335 4-24-75
0.053
<0. 025
0.042
0.094
0. 112
0. 113
0.088
0. 10
0. 1 5
0. 15
0.0248
April. 1975
Distance
(meters)
1300 U
800 U
1300 U
1300 U
800 U
1600 U
1600 U
0
0
0
0
2100 D
2100 D
1300 D
800 D
800 D
s (u / 31
Mercurv
0. 0028
0.0118
0. 0066
0. 0049
0. 0094
0.0119
0. 0061
0. 0069
0. 0076
0. 0035
0. 0085
0.0067
0. 0123
*
0. 008V
0. O0'*2
. 0. 0029
0. 0087
0. 0103
0. 0077
0. 0039
0. 0066
0. 0071
0. 0074
0. 0024
0. 0024
-------
TABLE 25. TRACE METAL CONCENTRATIONS IN PERIODS 4 AND 5
PARTICIPATE AIR SAMPLES
Trace Metal Concentrations
Location Date
Residential 2-9-76
Upwind 2-11-76
(>300m) 2-12-76
2-13-76
2-14-76
2-15-76
Geometric Mean
Arithmetic Mean
Standard Deviation
Sewage 2-9-76
Treatment 2-9-76
Plant 2-9-76
(<_300m) • 2-10-76
2-10-76
2-11-76
2-11-76
2-11-76
2-12-76
2-12-76
2-12-76
2-13-76
2-13-76
2-14-76
Z-15-76
2-15-76
2-15-76
2-15-76
2-16-76
2-16-76
Geometric Mean
Arithmetic Mean
Standard Deviation
esldential 2-10-76
Oownwind 2-11-76
(>300m) 2-13-76
2-14-76
2-14-76
2-15-76
2-15-76
Geometric Mean
Arithmetic Mean
Standard Deviation
Oetection Limit:
Distance
{ meters)
1600 U
500 U
3000 U
500 U
3300 U
500 U
240 D
1 D
125 D
1 D
230 D
100 D
100 D
1 D
150 D
190 D
1 D
150 D
1 D
1 D
190 D
1 D
230 D
230 D
100 D
100 D
1300 D
2100 D
3300 D
500 D
750 D
1300 D
3300. D '
Period 4
Cadmium
0. 0011
0.0008
0.0013
0.0055
0.0017
0.0008
0.0014
0.0019
0.0018
0.0018
0.0009
0.0008
0. 0013
0.0012
0. 0011
0. 0005
0. 0009
0. 0017
0.0011
0.0012
0.0046
0.0043
0.0020
0. 0006
0.0064
0. 0003
0.0021
0. 0014
0. 0045
0.0014
0. 0019
0.0017
0. 0009
0.0008
0. 0027
0. 0016
0. 0011
0.0011
0.0014
0.0013
0. 0014
0.0007
0. 0003
Lead
0. 29
0.27
0. 11
2.08
0.79
0.76
0.47
.0.72
0.72
0. 11
0.09
0. 11
0. 24
0. 21
0. 11
0. 07
0. 17
0. 15
0.43
0. 19
1.64
1. 75
0. 55
0.59
0.87
0. 59
1. 11
0. 59
0. 98
0.33
0.53
0. 51
0. 12
0.40
1. 92
0. 55
0. 73
0.67
0.78
0. 57
0.74
0. 57
0. 04
Mercury
#
0.022
0.016
0.034
0.082
0.045
0.034
0.040
0.026
*
*
0.077
t
0. 024
0. 026
**
t
0. 01 1
0. 036
t
0. 012
0. 215
**
0. 113
**
**
#*
0. 063
0.016
0. 038
0. 059
0.064
0. 016
0. 008
0. 015
0. 009
0. 014
0.011
t
0.0118
0. 0122
0. 0033
Date
9-30-76
10-1-76
10-4-76
10-6-76
10-7-76
9-30-76
9-30-76
10-1-76
10-1-76
10-2-76
10-2-76
10-3-76
10-3-76
10-3-76
10-4-76
10-6-76
10-6-76
10-7-76
10-7-76
10-1-76
10-2-76
10-2-76
10-3-76
10-3-76
10-4-76
10-4-76
10-6-76
10-6-76
10-7-76
10-7-76
0.002
Distance
(Meters)
800 U
800 U
1300 U
2200 U
2200 U
0
150 D
0
162 D
0
162 U
0
162 U
150 D
0
0
210 D
0
150 D
420 D
1300 D
3100 D
3100 D
1300 D
2200 D
840 D
1300 D
3300 D
540 D
4400 D
Period 5
Cadmium
0.0019
0.0037
0.0011
0.0007
0. 0008 .
0.0013
0.0016
0.0012
0.0018
0.0010
0. 0010
0.0015
0. 0030
0.0021
0.0013
0.0011
0. 0011
A
0. 0004
0.0009
0.0006
0.0011
0.0012
0.0013
0.0007 '
0. 0015
0.0033
0.0050
0. 0015
0.0024
0. 0014
0.0021
0. 0008
0.0005
0.0031
0. 0011
0.0017
0.0021
0.0013
0.0003
Lead
0. 13
3.23
0.61
2.32
4.73
1.23
2.20
1. 89
1.39
0.70
3.71
3.68
2. 51
1.81
1. 23
1. 11
0. 72
A
0. 13
0.22
0. 55
0. 56
0. 95
1.41
1.21
0.72
1.47
1.75
0.86
0.65
0.24
0. 15
0.26
0.30
0.85
1. 17
0. 59
0.77
0.53
0. 04
Mercur
0.044
0. 025
0.005
*
0.002
0.010
0.019
0, 020
0, 104
0.048
0.013
0.021
0.058
0.018
0. 017
0.016
0. 042
0.006
0. 003
tf
0. 003
<0. 001
0. 014
0. 027
0.029
0.044
0.015
0.021
0.007
*
0. 005
0.020
0. 019
0.006
0.002
0.036
0. 010
0.017
6.014
0.002
* Sample lost in shipment
f Off scale
** No sample taken
rT Tube broken in shipment
A Equipment failure
97
-------
PERIOD 4 CONCENTRATIONS
CADMIUM CONCENTRATIONS
IN AIR SAMPLES ug/m3
u
§
u
s
§
3000 2000 1000
METERS UPWIND
METERS DOWNWIND
PERIOD 5 CONCENTRATIONS
CADMIUM CONCENTRATIONS
IN AIR SAMPLES ug/m3
2000
METERS UPWIND
METERS DOWNWIND
Figure 13. Cadmium concentrations plotted by distance.
98
-------
PERIOD 4 CONCENTRATIONS
LEAD CONCENTRATIONS
IN AIR SAMPLES ug/m3
3000 2000 1000
METERS UPWIND
METERS DOWNWIND
PERIOD 5 CONCENTRATIONS
I i i I i
3000 2000 1000 0 . 0
METERS UPWIND
LEAD CONCENTRATIONS
IN AIR SAMPLES ug/m3
! • I I I I
1000 2000 3000 4000
METERS DOWNWIND
Figure 14. Lead concentrations plotted by distance.
99
-------
PERIOD 4 CONCENTRATIONS; ALL DATA
off
BC*le~
.067 -
MERCURY CONCENTRATIONS
IN AIR SAMPLES ug/m'
1000
METERS U PWIND
1000 2000 3000
METERS DOWNWIND
PERIOD 4 CONCENTRATIONS; OFF SCALE VALUES EXCLUDED
2000 1000
METERS UPWIND
1000 2000 3000
METERS DOWNWIND
(continued)
Figure 15. Mercury concentrations plotted by distance.
100
-------
PERIOD 5 CONCENTRATIONS
MERCURY CONCENTRATIONS
IN AIR SAMPLES ug/m3
.04-
METERS UPWIND
1000 2000 3000
METERS DOWNWIND
Figure 15. (continued).
101
-------
concerning the sewage treatment plant as a possible local source of aerosolized metal and a potential health
hazard in the residential downwind areas were considered through paired comparison procedures. The cad-
mium, lead, and mercury concentrations from Periods 4 and 5 were combined to increase the sample sizes.
The samples taken each day were obtained over approximately the same 24-hour periods, so the samples
taken at various distances from the plant were paired according to day.
No actual statistical test was performed on the cadmium data, as preliminary inspection of the means for
residential upwind, sewage treatment plant, and residential downwind samples, and of the plots of cadmium
concentrations with distance clearly indicated that there were no differences between the concentrations ob-
tained from these three general areas.
Initial inspection of the mean concentrations of lead suggested that the upwind residential concentrations
might generally be greater than the closest downwind concentrations. This suggests that other local sources of
aerosolized lead, such as automotive exhaust near expressways and highways, predominate over the sewage
treatment plant contribution to aerosol lead levels. A paired-sample t-test was conducted, comparing the
natural log transformed lead concentrations upwind (at least 300 meters from the plant) and the closest down-
wind concentrations for each day. This test found no significant difference between the concentrations ob-
tained upwind and at the sewage treatment plant.
Because many of the air samples taken at the closest downwind distances had missing or off-scale mer-
cury concentrations (see footnotes to Table 25), a geometric mean of all downwind samples taken at a
distance of no more than 300 meters was used as a comparison value against the upwind sample concentra-
tions for each day. A t-test was employed to test the differences between these paired samples; the resulting t-
value reached only borderline significance levels (p = .065), although it would have reached significance if a
one-sided test of significance had been employed.
Because it seemed likely that the off-scale values reported at distances of 1 meter downwind (actually
above the center of the first stage aeration basin) in Period 4 were true measures of the mercury concentra-
tions at that location, a sign test was also applied to the differences between upwind and aeration basin
samples, with these off-scale values included. This test yielded the same result as had the t-test, indicating no
difference at a two-sided .05 level of significance (p = .07). These two statistical tests suggest that no strong
inferences should be drawn concerning the sewage treatment plant as a source of aerosolized mercury without
obtaining additional confirming data. A comparison of the residential upwind concentrations against the
residential downwind levels (using the geometric mean of all samples taken beyond 300 meters on a given day)
was not significant. Thus, if the plant is indeed a source of mercury in the air, its contribution is not detec-
table in downwind residential areas.
Trace Metal Samp/ing Variability-^ with the microbiological aerosol sampling, a measure of trace metal
sampler variability was available from two samples taken on the same day and at the same distance. The
average coefficients of variation obtained were 0.88 for cadmium, 0.39 for lead, and 0.84 for mercury. Since
these coefficients are based on extremely limited data, they are imprecise measures of sampling variation. Ten
sets of duplicate aerosol lead data were taken using the same samplers on a neighborhood lead study/46^ This
data yielded an average coefficient of variation of 0.092 for lead sampling variability.
As with the microbiological air data, the trace metal sampler variability, large location variability, and
small sample sizes suggest that negative source and distance findings about trace metals indicate only that the
Egan plant does not make a predominant contribution to the downwind levels of the trace metals.
Soil Samples
Samples from the fifty sites were taken in each of the four periods for microbiological and trace metal
analyses. The microbiological analyses sought pathogenic bacteria and enteric viruses but also included stan-
dard plate count and such sewage indicators as coliforms and fecal streptococci. Trace metal concentrations
102
-------
were obtained for lead, copper, cadmium, zinc, and mercury. The results of these investigations were submit-
ted to statistical analysis according to the appropriate methods as outlined in the following methods section,
and the results and inferences drawn are presented below.
Statistical Methods-
The data from the microbiological and trace metal analyses are in the form of a matrix of four periods
and 50 sampling sites, for a total of 200 observations. The purposes of the statistical analysis were to deter-
mine the effects of the plant's operation through terms of pre-operational (Periods 1 and 2) vs. post-
operational (Periods 4 and 5) comparisons and to establish whether operational period increases have a
distance relationship. To accomplish this, a two-way analysis of variance (ANOVA) was used, with factors of
periods and sites. A no-interaction or additive model was selected, since no replications of the analyses were
made and there is thus no independent source of sampling error. The interaction between periods and sites is
assumed to be negligible so the residual sum of squares can be attributed to sampling error and used to
calculate the F- ratios for significance testing.
The method of orthogonal contrasts has been used to test the hypotheses concerning plant and distance
effects. When a prior F-test indicated that the periods factor was significant for a data set, two hypotheses
have been tested. The first investigated the pre-operational vs. post-operational levels of the variable, and the
second compared seasons for the operational Periods 4 (February 1976) and 5 (October 1976). The null
hypotheses may be stated as:
Ho, : n\ + /iz - /i4 - 1*5 =0
Hoz : fit - fit =0
To partition the periods sum of squares using this method requires that the contrasts be mutually orthogonal,
which is easily verified in this case. Each sum of squares calculated for a contrast has one degree of freedom
associated with it and the test of significance was made using an F-ratio, comparing the mean square for the
contrast to the error mean square.
Similarly, if there was a significant sites factor, a contrast was used to test for the effect of closeness to
the plant. Sites falling within or near the 500-meter radius around the plant are considered "near" for the
purposes of this analysis and the remainder are classified as "far." There are 18 near sites and 32 far ones.
This hypothesis may be stated as:
ere £/*N • sum °f tne "near1 ' means
£/tp- sum of the "far" means
and 16 and 9 have been selected so the sum of the coefficient equals zero, as required for a contrast. This con-
trast was also tested for significance using an F-ratio of the mean squares for the contrast and for error.
A possible effect resulting from the treatment plant operation was considered to be indicated when there
was a significant difference in the period terms, substantiated by the pre- vs. post-operational period con-
trasts, and when the concentration pattern with distance of the site from the plant was consistent with the
plant being a significant source.
Microbiological-
In all four periods, every soil sample analysis for pathogenic bacteria, enteric viruses, coliforms, and fecal
Streptococci was negative. All pathogenic bacteria were searched for, including Klebsiella pneumoniae, Pro-
teus, Pseudomonas, Salmonella, Shigella, Staphylococcus aureus, and streptococcus-beta. As a result, the
only microbiological data available to evaluate, the sewage treatment plant were the standard plate counts.
They are presented in Table 26 for the four periods under study.
103
-------
TABLE 26. STANDARD PLATE COUNT FOR SOIL SAMPLES
(no./mlX 106)
S;_
IIC
1
T
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
Period
1
98
280
20
190
740
47
150
2.3
14
10
79
69
2.7
50
310
270
420
890
52
220
71
41
49
50
80
2
160
8.5
17
8.1
1.7
26
52
64
290
230
8.8
5.5
3.4
24
6.6
4.5
3.4
13
3.7
67
20
3.8
12
0.2
8.0
4
3.3
1.6
120
27
17
20
140
10
8.6
50
4.5
22
0.15
7.1
11
.50
3.6
9.4
0.47
4.6
5.3
2.9
13
6.0
2.1
5
80
200
620
4600
50
30
100
90
130
60
40
90
6.3
• 8.4
6.1
6.0
0.15
22
20
8.0
9.1
14
6.8
1.8
5.8
Sift«
uc
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Period
1
290
470
110
1.0
8.9
6.0
140
35
490
62
61
500
250
540
360
290
57
15
48
41
42
270
0.70
5.3
150
2
14
30
12
14
7.0
8100
86
8700
910
40
37
32
19
18
10
7.5
8.9
1.8
7.1
11
720
8.3
1.7
3.9
6.7
4
3.0
18
2.8
2.8
2.3
1.5
6.1
4.0
3.2
6.7
0.43
5.0
2.3
1.1
4.7
0.89
1.5
4.4
4.0
8.8
4.1
3.2
3.3
6.6
2.0
5
13
6.4
7.2
12
3.1
7.7
6.4
6.3
5.4
4.1
3.0
21
6.9
3.4
30
2.6
10
0.70
5.5
I.I
3.6
9.3
7.8
0.096
2.6
As can be seen, the results were quite variable, and those from Period 1 were higher than those of suc-
ceeding periods by one to two orders of magnitude. On the basis of the observed counts and on previous ex-
perience with microbiological data, it was decided to use a logarithmic transformation (In x) prior to con-
ducting the ANOVA on the data. The results of the ANOVA are contained in Table 27.
TABLE 27. ANOVA ON STANDARD PLATE COUNT IN SOIL SAMPLES
OnX)
Source
Periods
I,2vs4, 5
4 vs 5
Sites
Near vs Far
Period X Site
(error)
Total
df
3
1
1
49
1
147
199
SS
177.81
122.46
15.21
192.73
2.85
394.82
765.36
MS
59.27
122.46
15.21
3.93
2.85
2.69
F
22.03
45.52
5.65
1.46
1.06
Significance
Level
<0.01
<0.01
<0.01
0.05
N.S.
Period Means (In X)
1245
4.20 2.93 1.61 2.39
Site Means (In X)
Near Far
2.74 2.81
The periods factor is significant beyond the 1-percent level, which means that some changes did occur in
the observed levels from period to period. The .period means as shown in the table indicate that the highest
levels were in Period 1 while the lowest occurred in Period 4, the same season of the year two years later. The
104
-------
contrasts of interest with respect to periods were tested and both the pre- vs.post-operational contrast and the
Period 4 vs. Period 5 contrast were significant beyond the 1-percent level.
The pre-post comparison indicates that there was a change, and that the overall levels decreased
significantly. The comparison of Periods 4 and 5, however, gives a significant increase for the October sampl-
ing period. The geometric means are 5.0 x 106, and 10.9 x 10' respectively, or a factor of two between the
two periods.
The sites factor was also significant, and the contrast of near sites vs. far sites is tested for the plate count
data. The criterion for a near site is one within a 500-m radius from the center of the first stage aeration basin.
The F-ratio for this test is not significant at the 10-percent level and thus no effect related to distance from the
plant can be observed.
The interaction sum of squares is used to represent the sampling error term, under the assumption that
no interactive effect is present. For these data, presence of an interaction would indicate that some stations in-
creased while others decreased in significant amounts from one sampling period to another, rather than a
general trend up or down for an entire sampling period. In this ANOVA, the magnitude of the error mean
square is small enough to imply that no interactive effect was present which might have unduly influenced the
results.
Thus, a decrease in standard plate count was observed in the post-operational periods versus the pre-
operational, and an increase occurred from Period 4 to Period 5. However, the sites comparison did not
indicate that those sites close to the plant were higher or lower on average than those some distance away from
the plant. As a result, no influence of the plant on these period-to-period changes in the observed standard
plate count can be detected, and they are attributed instead to other factors.
Trace Metals-
The soil samples were analyzed for five trace metals— lead, copper, cadmium, zinc and mercury—during
periods 1, 2, 4 and 5. The results from these analyses are used in the ANOVA model described in the
preceding Statistical Methods section to determine if any increase in the environmental levels of these metals
occurred subsequent to the plant beginning operation and, if so, whether these changes could be related to
closeness to the plant. Fifty sites were used to collect samples during each period, and the analyses were com-
pleted for all metals except for zinc in Period 5.
The ANOVA model requires homogeneous variance among the factors, so a preliminary investigation
was made into the error terms. Using the variability among sites within a given period as the standard, stan-
dard deviations were calculated and used to calculate Bartlett's test statistics for each metal. Comparing these
statistics to a chi-square distribution gives the significance levels shown in Table 28. To achieve homogeneity,
the results were transformed using a natural logarithmic transformation. For those metals which had values
between 0 and 1, a constant of 1 was added to the value prior to taking the logarithmic transformation. This
results in only positive values being used and reduces the overall variability. The standard deviations and
Bartlett's statistics are presented for these transformed data as well.
As can be seen, for none of the metals could the variances be considered homogeneous for the non-
transformed data. Under the transformation, however, all but the Hg and Cd data achieved homogeneity,
and the ANOVA is conducted on all metals under the transformation. The rationale behind this is that the
transformed values showed a greater agreement than the nontransformed, that the magnitude of the dif-
ference between the one high value and the remainder is small, and that this provides greater consistency in
the results presented. Since the significance level of the F-ratio is affected by violating this assumption, cau-
tion is then exercised in interpreting the results of any significant factors in the mercury or cadmium analyses.
The results of the ANOVA's and subsequent contrasts of sub-hypotheses, if any, are presented below for each
metal under study.
105
-------
TABLE 28. EQUALITY OF VARIANCE TESTS-ORDINARY VS TRANSFORMED
Metal
Lead
Copper
Cadmium
Zinc
Mercury
Period
1
2
4
5
1
2
4
5
1
2
4
5
1
2
4
1
2
4
5
Std Dev
14.31
15.56
37.65
68.99
0.77
0.55
1.07
1.11
0.45
0.34
0.22
0.51
11.62
17.62
10.50
0.04
0.09
0.02
0.03
Ordinary
Test Significance
Statistic Level b W °
0.60
0.81
0.62
151.24 <0.01 0.71
0.30
0.23
0.36
27.76 <0.01 0.31
0.21
0.17
0.14
34.72 <0.01 0.25
0.74
0.82
15.48 <0.01 0.64
0.03
0.07
0.02
133.37 <0.01 0.02
Transformed*
Test Significance
Statistic Level
5.64 N.S.
9.50 0.025
17.98 <0.01
2.96 N.S.
114.27 <0.01
*ln X for zinc, In (1 + X) for others.
Lead-The analyses for lead content are summarized in Table 29. The ANOVA for the lead results from
the four periods is presented in Table 30. As can be seen, the F-ratios for both periods and sites are significant
beyond the one-percent level and contrasts were used among the means to determine if relevant differences ex-
ist.
For the periods factor, the two contrasts used were those representing a change from the pre-operational
to the post-operational period and for contrasting Periods 4 and 5. The sums of squares for the contrasts,
each with one degree of freedom, were partitioned out of the total sums of squares for periods and F-ratios
were calculated.
There was a significant increase in the soil lead from the pre- to post-operational periods, as can be seen
from the F-ratio for that contrast. The significance level of the pre- vs. post-contrast is less than 0.01, and the
difference of 0.714 translates into 2.04^g/g increase in the original scale. The Period 4 vs. Period 5 contrast
was also significant beyond the 1-percent level, with a calculated difference of 0.463 in the transformed scale.
This represents a difference of 1.59 /tg/g in the average lead content between these two periods.
To determine if the observed increase could be related to the operation of the plant, the sampling sites
were investigated. If the increase in lead results from materials passed through aerosols, then the lead content
of the sites close to the plant should be higher than that of the more distant sites. The criterion for near vs. far
is selected to be within or near a 500-m radius around the plant. Using this, 18 sites fell into the near and the
remaining 32 into the far category. The sums of squares for this contrast was partitioned out of the sites fac-
tor and the F-ratio is significant beyond the 1-percent level.
The transformed mean of the 18 near sites is 2.721 vs. 3.057 for the 32 far sites, however, and the far sites
had higher average lead content in the soil than the near sites. As a result, there is no indication that the in-
crease is related to the plant.
106
-------
TABLE 29. DETERMINED CONCENTRATIONS OF LEAD
IN SOIL SAMPLES
(iug/gm, dry weight)
Site
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
IS
19
20
21
i'2
23
24
25
Period
1
98
280
20
190
740
47
150
2.3
14
10
79
69
2.7
50 '
310
270
420
890
52
220
71
41
49
50
80
2
160
8.5
17
8.1
1.7
26
52
64
290
230
8.8
5.5
3.4
24
6.6
4.5
3.4
13
3.7
67
20
3.8
12
0.2
8.0
4
3.3
1.6
120
27
17
20
140
10
8.6
50
4.5
22
0.15
7.1
11
50
3.6
9.4
0.47
4.6
5.3
2.9
13
6.0
2.1
S'
80
200
620
4600
50
30
100
90
130
60
40
90
6.3
8.4
6.1
6.11
0.15
22
20
8.0
9.1
14
6.8
1.8
5.8
Site
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Period
1
33.40
69.05
10.35
14.75
31.29
12.61
12.87
20.63
17.02
10.93
13.52
9.00
29.05
33.61
7.37
10.38
19.19
17.48
12.68
11.68
14.12
13.30
20.44
26.22
16.03
2
36.36
78.63
4.04
6.32
65.89
13.35
13.98
20.86
61.95
17.94
18.57
9.39
27.54
29.83
1.70
7.77
14.91
10.92
8.61
7.98
7.98
15.13
14.49
23.31
13.23
4
38.27
47.24
15.55
13.46
23.92
12.89
11.43
20.50
39.85
24.91
14.94
13.48
17.28 •
49.80
9.96
11.00
44.86
15.05
11.86
10.42
13.60
21.41
14.18
14.76
10.42
5
123.82
88.94
52.45
21.10
57.31
17.58
19.86
55.70
50.02
21.50
18.72
17.42
26.53
19.37
13.02
13.82
18.38
17.88
13.b5
14.47
56.50
21.30
15.77
20.16
18.53
TABLE 30. ANOVA ON LEAD IN SOIL SAMPLES
(InX)
Source
Periods
1, 2vs4,5
4vs5
Sites
Near vs Far
Period X Site
(error)
Total
df
3
1
1
•)
1
1 •'•
199
SS
12.3933
6.3725
5.3592
67.7352
2.85
25.6417
105.7702 .
MS
4.1311
6.3725
5.3592
1.3824
2.85
0.1744
F
23.69
36.54
30.73
7.93
16.34
Significance
Level •
<0.01
<0.01
<0.01
<0.01
<0.01
Period Means (In X)
1245
2.839 2.676 2.883 3.346 ,
Site Means (In X)
Near Far
.2.721 3.057
107
-------
Since there is no replication term in these data, the period-by-site interaction is assumed to be zero and
used as the error term. If the assumption were not correct, there could be an increase in the near sites not ac-
countable for by the increase over periods and not observed at the far sites. If this were so, one would expect a
relatively large mean square for this interaction term. However, the mean square is of relatively low
magnitude in this ANOVA, and there is no indication of an unaccounted for interaction in these data. Thus,
the increase in soil lead content is not attributed to the operation of the treatment plant.
Copper~The analytical results for copper are shown in Table 31, and the results of the ANOVA on these
data are contained in Table 32. The F-ratio for periods was insignificant at all normal criteria, with an
estimated significance level of 20-25 percent. The sites factor was significant beyond the one-percent level.
TABLE 31. DETERMINED CONCENTRATIONS OF COPPER
IN SOIL SAMPLES
(/jg/gm, dry weight)
Site
1
•>
3
4
5
6
7
8
9
10
11
n
13
14
15
16
17
18
19
20
21
__
23
24
25
1
l.5u
.63
.63
.22
.19
0.93
0.97
0.81
0.73
0.44
0.86
0.73
1.52
0.41
0.92
1.36
4.44
0.82
0.43
0.41
0.47
0.51
3.66
3.97
0.90
PC
t
0.56
1.52
1.29
1.07
0.97
0.92
0.88
0.79
0.97
0.97
1.09
0.87
1.72
0.46
1.77
2.27
4.09
0.78
0.50
0.50
A «Q
0.53
1.87
1.90
0.98
iod
4
3.89
1 .00
3.06
7.22
0.59
0.87
0.72
0.93
0.46
0.46
0.59
0.43
3.14
0.59
0.87
1.38
1.97
0.74
0.43
0.28
u.4i
0.42
0.84
0.93
0.90
5
0.76
1.32
1.40
1.32
1.18
0.82
0.97
0.97
1.40
0.39
1.08
1.01
1.57
0.80
1.63
1.22
8.67
1.36
0.87
0.52
0.67
0.67
2.08
0.93
1.13
Sire
26
21
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
-15
46
47
48
49
id
Period
1
0.37
0.37
1.14
0.69
0.87
0.94
0.91
0.84
0.88
0.94
.01
.80
.79
.66
.50
.11
.00
.18
.34
.6u
.87
.79
.81
0.99
0.88
i
0.40
0.36
1.42
1.1!
0.67
0.96
0.78
1 .00
l.W
0.87
0.82
0.82
0.64
1.18
1.68
0.88
1.06
0.88
1.50
1.37
1.41
1.10
0.74
0.51
3.83
4
0.39
0.36
0.70
0.86
0.70
0.74
1.26
0.96
0.84
1.72
0.61
0.79
0.70
1.90
1.93
.82
.54
.87
.61
.87
.21
.12
.34
0.73
3.58
5
0.47
0.53
0.53
1.34
1.07
0.96
1.28
0.96
0.49
0.96
0.89
1.48
0.76
1.80
2.59
1. 12
1.59
1.46
4.10
2.58
1.39
0.99
1.06
0.99
2.11
TABLE 32. ANOVA ON COPPER IN SOIL SAMPLES
Source
Periods
Sites
Period X Site
(error)
Total
df
3
49
147
199
SS
0.2364
12.7862
7.1656
20.1882
MS
0.0788
0.2609
0.0487
F
1.62
5.36
Significance
Level
>0.20
<0.01
Period Means (In (1 + X)l
1245
0.749_ 0.698 0.724 0.792
108
-------
From the ANOVA and the means shown in the table, it is concluded that there were no changes in the
copper content of the soil over the period of time under study. The range from lowest to highest average con-
centration over the four periods was only from 1.01 to 1.21 jig/g in the original scale, and no linear trend in
the data can be observed.
As in the lead data, the mean square for interaction indicated that this was a valid source for the error
term and that no unidentified interaction existed which complicated the results or conclusion.
Cadmium-Jhe results of the analyses for cadmium content in soil samples are contained in Table 33. The
summary of the ANOVA on these data is shown in Table 34 for the cadmium analyses over the four periods.
The F-ratio for periods was significant at the 1-percent level and two sub-hypotheses were tested. The sites
factor was also significant at the 1-percent level and the near vs. far sites hypothesis was tested for these data
as well.
The period means as shown in the table indicate where the differences lie which are indicated by the F-
ratio. The pre-operational means are of equivalent magnitude, but the post-operational winter mean is
significantly lower while the fall mean is higher than the pre-operational means. The test for a pre- vs. post-
effect, as a result, is not significant, with an estimated significance level of 20 percent. The Period 4 vs. Period
5 contrast gives a resulting F-ratio which is significant beyond the one-percent level. The transformed mean
for Period 5 shown in the table represents an observed concentration of 0.47 ^g/gm in the original scale.
Since there was a significant increase in Period 5, the site means were examined to see if the high concen-
tration could be related to proximity to the plant. The contrast for near vs. far difference yields an F-ratio
which is not significant, however. This implies that near and far sites had the same average observed levels. As
a result, there is no indication that the rise in soil cadmium level is related to the plant operation.
The mean square for interaction represents the sampling error for these data. If the assumption of no in-
teraction is incorrect, then this would be expected to be relatively large. As can be seen, however, this mean
square is small compared to those for the period and site factors and the assumption is justified. The conclu-
sion, then, is that no increase in soil cadmium was observed which can be attributed to the plant operation.
TABLE 33. DETERMINED CONCENTRATIONS OF CADMIUM
IN SOIL SAMPLES
. (;ug/gm. dry weight)
Site
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
IS
19
20
21
22
23
24
25
Period
1
0.25
0.93
0.52
0.21
0.91
0.29
0.17
0.25
0.31
0.28
0.32
0.24
0.21
0.41
0.21
0.77
0.20
0.35
0.28
0.76
0.63
0.75
4.28
0.16
0.28
->
0.40
0.36
0.83
0.23
0.83
0.35
0.28
0.33
0.29
0.13
0.32
0.27
0.28
0.49
0.29
0.27
0.24
0.40
0.41
0.63
0.69
0.69
3.34
0.20
0.28
4
0.09
0.24
0.31
0.43
0.56
0.25
0.17
0.20
0.20
0.19
0.23
0.32
0.12
0.23
0.19
0.75
0.15
0.23
0.20
0.55
0.56
0.35
2.04
0.31
. 0.25
5
0.81
0.39
0.57
0.68
0.49
0.44
0.24
0.51
0.29
0.41
0.28
0.48
0.23
0.49
0.21
0.97
0.41
0.31
0.36
0.97
0.87
0.43
5.05
•0:37
0.37
Sile
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Period
1
0.56
0.55
0.52
0.31
0.17
0.31
0.25
0.17
0.15
0.13
0.16
0.19
0.45
0.27
0.23
0.28
0.53
0.68
0.52
0.20
0.44
0.45
1.17
0.19
0.27
2
0.56
0.57
0.15
0.17
0.39
0.36
0.25
0.16
0.52
0.27
0.24
0.23
0.57
0.56
0.31
0.35
0.56
0.35
0.33
0.24
0.33
0.32
0.44
0.37
0.72
4
0.56
0.39
0.41
0.16
0.13
0.33
0.21
0.19
0.27
0.23
0.12
0.13
0.29
0.28
0.13
0.20
0.32
0.31
0.19
0.21
0.27
0.60
0.21
OJ7
0.13
5"
1.03
0.63
0.89
0.16
0.27
0.32
0.41
0.36
0.31
0.23
0.27
0.27
0.44
0.55
0.25
0.29
0.47
0.41
0.33
0.32
0.35
0.57
0.43
0.29
0.35
109
-------
TABLE 34. ANOVA ON CADMIUM IN SOIL SAMPLES
Source
Periods
1, 2vs4,5
4 vs5
Sites
near vs far
Period X Site
(error)
Total
df
3
1
1
49
1
147
199
SS
0.4523
0.0177
0.2175
7.7381
0.0007
1.4726
9.6630
MS
0.1508
0.0177
0.2175
0.1579
0.0007
0.0100
F
15.08
1.77
21.75
15.79
<1
Significance
Level
<0.01
~0.20
<0.01
<0.01
N.S.
Period Means fln(l+X)J
1245
0.339 0.340 0.255 0.387
Mercury-The mercury analyses on soil samples are contained in Table 35. These were used in the
ANOVA model above to obtain the ANOVA summary in Table 36. The periods factor was not significant for
these data, with an approximate level of 10 percent. The period means as shown in the table further suggest
that no general increase occurred in environmental levels of Hg in soil. The significance of the site factor is to
be expected, but need not be considered in view of the lack of significance of the period term.
As with the previous data, the magnitude of the mean square for the interaction suggests that the
assumption of an additive model is correct. It is concluded, then, that no observable rise in the Hg levels in
soil could be attributed to the plant operation.
TABLE 35. DETERMINED CONCENTRATIONS OF MERCURY
IN SOIL SAMPLES
(jug/gm, dry weight)
Site
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
Period
I
0.04
0.09
0.04
0.04
0.05
0.04
0.06
0.03
0.04
0.06
0.04
0.03
0.03
0.04
0.03
0.10
0.02
0.04
0.03
0.06
0.10
0.08
0.27
0.03
0.04
2
0.04
0.04
0.07
0.03
0.11
0.04
0.05
0.04
0.04
0.05
0.04
0.04
0.04
0.05
0.03
0.04
0.04
0.05
0.19
0.05
0.06
0.07
0.10
0.05
0.03
4
0.04
0.04
0.06
0.10
0.04
0.04
0.04
0.04
0.04
0.06
0.04
0.04
0.03
0.03
0.03
0.13
0.03
0.04
0.03
0.04
0.06 .
0.04
0.11
0.04
0.03
5
0.10
0.05
0.05
0.05
0.04
0.04
0.04
0.05
0.05
0.10
0.05
0.07
0.05
0.05
0.04
0.15
0.07
0.06
0.05
0.08
0.08
0.07
0.13
0.06
0.04
Site
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Period
1
0.08
0.09
0.05
0.05
0.05
0.05
0.05
0.05
0.04
0.04
0.04
0.04
0.07
0.05
0.04
0.04
0.04
0.04
0.04
0.04
0.05
004
0.05
0.06
0.03
2
0.10
0.61
0.03
0.03
0.10
0.03
0.02
0.03
0.03
0.03
0.03
0.03
• 0.04
0.03
0.03
0.03
0.16
0.04
0.03
0.03
0.03
0.17
0.03
0.03
0.03
'4 '
0.06
0.06
0.03
0.03
0.04
0.03
0.02
0.04
0.03
0.03
0.04
0.03
0.04
0.03
0.03
0.03
0.04
0.04
0.04
0.03
0.03
0.05
0.04
0.03
0.03
5'
0.09
0.08
0.07
0.04
0.06
0.05
0.04
0.06
0.05
0.04
0.06
0.04
0.07
0.04
0.04
0.05
0.04
0.05
0.05
0.05
0.08
0.05
0.05
0.05
0.05
110
-------
TABLE 36. ANOVA ON MERCURY IN SOIL SAMPLES
Source
Periods
Sites
Period X Site
(error)
Total
df
3
49
147
199
SS
0.0091
0.1339
0.1931
0.3361
MS
0.0030
0.0027
0.0013
F
2.31
2.08
Period Means [In (1 +XjJ
1245
0.048 . 0.057 0.039 0.055
Significance
Level
0.10
<0.01
Zinc-The analytical results for zinc in soil are shown in Table 37, and the summary of the ANOVA on
these data in Table 38. Analyses on the soil samples for zinc content were made only on samples from the first
three periods. The periods factor in the ANOVA proved to be insignificant for these data, showing less
variability than the interaction term. The period means shown represent a range of only 13.28 to 13.82 mg/g
average concentration from low to high.
As will be noted, the interaction term is relatively high and the possibility exists that the no-interaction
model is not justified for these data. As a result, the hypothesis of near sites equal to far sites is tested in spite
of the lack of an observable period effect. The F-ratio for this hypothesis is much less than one, and no dif-
ference can be found between those close to the plant and those farther away.
TABLE 37. DETERMINED CONCENTRATIONS OF ZINC
IN SOIL SAMPLES
(lug/gm, dry weight)
Site
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
Period
1
4.31
8.72
10.11
12.93
11.73
6.51
11.93
4.51
12.93
14.27
6.98
16.24
8.76
17.22
3.83
32.95
17.22
50.12
11.32
23.32
15.83
39.04
42.73
6.00
6.03
2
11.55
10.56
17.28
7.80
10.76
18.26
14.71
28.92
13.72
3.43
7.55
17.35
7.55
21.46
9.51
9.12
14.80
83.32
17.74
22.25
12.25
43.42
55.65
3.04
8.33
4
38.75
6.95
39.48
42.14
11.21
7.19
11.45
5.06
13.33
14.04
8.15
31.88
8.15
10.24
4.65
12.33
18.15
10.24
10.71
13.96
28.87
26.78
36.31
5.54
7.58 '
Site
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Period
1
26.42
30.18
8.60
6.23
32.35
9.20
9.59
14.55
4.45
2.47
6.03
7.62
41.85
32.55
9.21
43.71
34.60
20.71
21.51
16.95
20.71
22.70
42.92
16.16
14.57
1
29.71
28.35
3.02
4.77
36.72
7.70
9.65
10.62
36.92
4.39
6.14
3.61
40.27
22.50
101.76
16.37
22.21
25.91
10.91
4.67
5.46
9.74
10.32
15.58
6.82
4
46.08
23.37
13.93
8.27
15.55
8.70
16.84
13.90
20.84
15.70
8.03
7.12
32.69
25.13
6.89
6.55
32.19
11.82
9.53
8.60
8.83
37.48
7.69
14.11
5.97
111
-------
TABLE 38. ANOVA ON ZINC IN SOIL SAMPLES
(InX)
Source
Period
Site
Near vs Far
Period X Site
(error)
Total
df
2
49
1
98
149
SS
0.0423
46.7569
0.0954
33.5337
80.3329
MS
0.0212
0.9542
0.0954
0.3422
F
<1
2.79
<1
Significance
Level
N.S.
<0.01
N.S.
Period Means (In X)
1 2 4
2.626 2.586 2.600
As for the previous metals, the conclusion regarding the environmental levels of zinc in soil samples is
that no effect of the plant's operation could be detected on the observed concentrations.
Findings-
The findings from the statistical analyses on soil sample analytical data are summarized in Table 39. Only
one of the parameters, lead, showed an increase in the post-operational period and for this the distance com-
parison showed that the far sites were higher than those close to the plant. As a result, none of the soil data in-
dicated that there was a relationship between the results obtained and the operation of the sewage treatment
plant.
TABLE 39. STATISTICAL FINDINGS SUMMARY-SOIL SAMPLES
Seasonal Effect
(Winter vs Fall)
Statistical Inferences
Operational Effect
(Baseline vs Operational)
Distance Effect
(Near vs Far)
Indicated Relation-
ship to Sewage
Treatment Plant
Microbiological
Standard Plate Count
Fall increase
Operational decrease
No difference
Trace Metals
Cadmium
Copper
Lead
Mercury
Zinc
Fall increase
No difference
Fall increase
No difference
No difference
No difference
No difference
Operational increase
No difference
No difference
No difference
Not done
Higher at far sites
Not done
No difference
None
None
None
None
None
None
Water Samples
Samples from the 10 sites (ponds and streams) were taken for microbiological and trace metal analyses.
The microbiological analyses sought pathogenic bacteria and enteric viruses, but also included such
sewage indicators as coliforms and fecal Streptococci. The concentrations of the trace metals, lead, copper,
cadmium, zinc, and mercury, were also determined for each water sample. The results of these analyses and
the conclusions obtained from the data analysis are presented below.
112
-------
Statistical Methods-
The data bases for the water analyses take the form of a four by ten matrix of results, with four periods
and ten sampling sites. The statistical analysis for these follows the same general procedure as used in the soil
analyses. An ANOVA model is used with factors of periods and sites, and the assumption of additivity is
made; that is, that no interaction exists between periods and sites. Thus, the residual sum of squares after ac-
counting for period and site difference is assumed to be due to sampling error and is used as the basis for the
denominator in the F-tests of significance.
The effect of the treatment plant operation on the water quality is estimated using orthogonal contrasts
among the periods and sites as in the soil. If a significant periods term is noted, then two hypotheses are
tested:
Hoi : 11, + pi -
Ho2: Ht -
The first tests for an overall change from the pre- to the post-operational period and the second tests for a
change from the winter to fall period. Sums of squares for the contrasts are partitioned out of the periods sum
of squares with one degree of freedom each. The test for significance is an F-ratio compared to the error mean
square. For any contrast for which a significant effect is noted, the direction of the difference is noted and
conclusions drawn.
To support any findings which indicate that the treatment plant operation may impact on the environ-
ment, a third hypothesis is used. If the periods factor is significant, then the hypothesis of site differences due
to closeness to the plant is tested. For the purposes of this study, sites falling in and around the 500-m radius
of the plant are considered "near" and the remainder of the sites "far." There are four near sites and six far
ones using this criterion, and the hypothesis becomes:
where
E/iN - sum of the near site means
E/ip - sum of the far site means
and 3 and 2 are used to bring the sum of the coefficients to zero, as required for a contrast.
The treatment plant might be implicated as a source when the baseline vs. operational period contrast is
significant and the result is possibly attributable to the plant by a near site vs. far site comparison. If no
significant site factor is observed or if the site contrast is not significant, then the period result will be at-
tributed to a general increase or decrease in the studied variable unrelated to the treatment plant.
Microbiological-
Despite a thorough screening, no pathogenic bacteria or enteric viruses were found in any of the water
samples from any sampling period. All pathogenic bacteria were sought, including Klebsiella pneumoniae,
Proteus, Pseudomonas, Salmonella, Shigella, Staphylococcus aureus, and Streptococcus-beta. Since no fecal
coliforms were found in any water sample, the levels of fecal coliforms were below the analytical detection
limit of 1 MPN/lOOml. Quantitative determinations were obtained for total coliform and standard plate
count. These data are analyzed below.
113
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The total coliform concentrations in MPN/100ml are presented in Table 40 for the water samples. E. coli
and enterobacter were identified as predominant coliforms present. There is a slight increase pairwise between
the corresponding season periods. From October 1974 to October 1976, the observed values increased to 161
for all sites. From February 1975 to February 1976, there was an increase in the number of sites showing a
positive response, with two sites at the 161 level.
Since MPN data are only semiquantitative, a sign test has been used to assess these values in relation to
the possible effects of the treatment plant. To obtain sufficient data, the sites were compared seasonally, i.e.,
Period 1 vs. Period 5 and Period 2 vs. Period 4, and the results pooled to test for significance. The 20 pairs
thus generated show differences from pre- to post-operational of nine increases, one decrease, and 10 no-
change. Using the convention that no-change comparisons be deleted from the data set, the result is a sample
of size 10 with nine pluses and one minus. From a table of the distribution of the test statistic, the significance
level of this result is approximately 0.02, and is significant at the 5-percent level. Thus, there is some indica-
tion of a change in the post-operational periods, although the overall levels are quite low.
To assess the effect of the plant on these changes, the distance of the site from the plant is considered.
Sites 1, 3, 4, and 5 can be considered "near" sites, falling at or near the 500-m circle from the first stage aera-
tion basin. Of the increases observed, three of the nine were at near sites and the remaining six at the far.
Percentage-wise, this is 3/8 = 0.375 vs. 6/12 = 0.50, and thus there is no tendency for the increases to be
related to the closeness to the plant. There is no evidence, then, that the increases in the total coliform levels
seen in the post-operational period are related to the sewage treatment plant.
The standard plate count data are shown in Table 41, and provide the only data from all sites and
periods. As with the soil samples, the data from the first period are higher than those from the subsequent
periods by over an order of magnitude in many cases. It is not clear whether this reflects elevated shipping
temperature or actually represents higher bacterial levels. In any event, it dominates the statistical analyses.
Due to the skew nature of the plate count data, the data were first transformed using a natural
logarithmic transformation prior to running the ANOVA. This helps reduce the anomalous nature of the first
period's data. The ANOVA is summarized in Table 42.
The periods factor is significant beyond the one-percent level, as could be expected from the fact that
such high values were reported for period 1. The pre-operational vs. post-operational contrast was signifi-
cant, as could be expected, but the comparison between Periods 4 and 5 proved nonsignificant. To further
evaluate these results, a Least Significant Difference (LSD) statistic was calculated for these data. The LSD
allows pairwise comparisons of means at a given significance level to determine which are equivalent and
which are different. The LSD for this test is calculated as:
TO—
LSD.05 = t.05(27) ' E
where
105(27) -1-value ata=0.05 with 27 df
MSE - mean square for error from the ANOVA with 27 df
n - number of observations in the sample
Substituting gives
/3.350
LSD. os = (2.052)^
= (2.052)(0.579)
= 1.19.
114
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TABLE 40. TOTAL COLIFORM FOR WATER SAMPLES
(MPN/lOOml) •
Site
Period
1
2
3
4
5
6
7
8
9
10
-------
Thus, two means must differ by more than 1.19 to be considered separable at the 0.05 level. The range,
however, for the means except for Period 1 is only 1.446 - 0.557 = 0.889 and no difference is detectable. The
conclusion, then, is that only Period 1's value can be considered different, and that the plate count data do
not indicate any effect on the environment due to the operation of the treatment plant.
Trace Metals-
The water samples were analyzed for lead, copper, cadmium, zinc, and mercury content during Periods
1, 2, 4, and 5. The results from these analyses are used to evaluate the effect of the plant's operation on en-
vironmental levels of these metals. Where appropriate, the ANOVA model described in the water sample
statistical methods section is used for this evaluation. In other instances, the data are inadequate and an
evaluation is used based upon the mean values calculated for each period.
In the water samples, there are frequent observations which fall below the detection limit of the method.
For these, a value equal to one-half the detection limit is used in the statistical analyses. The rationale behind
this is that this would be the average expected value of these results if they are nearly equally likely and that
the value of the detection limit is sufficiently low that this could not be expected to unduly influence the
results.
The sample from site 1, Period 1 was analyzed in duplicate or triplicate on the majority of the metals. For
this value, the average of the results is used in succeeding analyses, while the tables reflect all of the analyses
made. The results of the statistical analysis are presented below for each metal under study.
Lead-The results of the analyses for lead in water samples are shown in Table 43 and are used to obtain
the ANOVA in Table 44. The results are multiplied by 104 for ease of computation and presentation. As can
be seen, the period factor is not significant at the 5-percent level, with an estimated significance of 0.13. The
sub-hypotheses for the period term are partitioned from the sum of squares since there is a marginal pro-
bability of a period effect. As can be seen, however, the contribution of the sum of squares due to a pre- vs.
post-operational effect or a Period 4 vs. Period 5 effect was minimal, and no significant variation could be
detected.
The sites term was clearly nonsignificant at any reasonable level, and no partitioning was made of this
term. However, the error mean square was of sufficient magnitude to suggest that there may have been an
interactive effect and that the sum of squares may not be attributable merely to sampling error. This
hypothesis is tested using Tukey's test of nonadditivity/47* which evaluates the validity of the assumption of
additivity in the model. A sum of squares for nonadditivity is partitioned from the error sum of squares and
TABLE 43. LEAD CONTENT OF WATER SAMPLES
(mg/S)
Period
Station
2
3
4
5
6
7
8
9
10
*N.D.-none detected. Detection limit: 0.0027 mg/C.
1
0.0056
0.0041
0.0044 .
N.D.*
0.0047
0.0045
N.D.
0.0051
N.D.
0.0028
0.0059
0.0059
2
0.0197
0.0042
0.0154
0.0207
N.D.
0.0088
0.0095
0.0241
0.0049
N.D.
4
0.0069
N.D.
0.0059
0.0069
0.0058
0.0087
0.0030
N.D.
0.0310
0.0095
5
0.0136
0.0057
0.0163
0.0145
N.D.
N.D.
N.D.
0.0049
N.D.
N.D.
116
-------
TABLE 44. ANOVA ON LEAD IN WATER SAMPLES
(mg/mlX 10-")
Source
Periods
I,2vs4, 5
4 vs 5
Sites
Period X Site
(error)
Nonadditivity
Remainder
Total
df
3
1
1
9
27
1
26
39
SS
2.7921
0.0073
0.1711
4.7869
11.8612
0.2019
11.6593
19.4402
MS
0.9307
0.0073
0.1711
0.5319
0.4393
0.2019
0.4484
F
2.12
<1
<1
1.21
<1
Significance
Level
0.13
N.S.
N.S.
N.S.
N.S.
Period Means (mg/ml)
1245
0.0038 0.0110 0.0081 0.0062
tested for significance by an F-ratio against the mean square of the remainder of the sum of squares. For this
ANOVA, the mean square is less than one and the additivity is determined to be a valid assumption.
The conclusion, then, is that no increase in environmental levels of lead in water was observed after the
plant became operational.
Copper-Analyses were made on the water samples for copper content only on the first two periods
samples. As a result, the results are summarized by mean and standard deviation only. The data and summary
statistics are contained in Table 45. The mean for Period 1 was 0.0085 mg/l, while Period 2 showed an
average of 0.0243 mg/^, roughly three times as large.
No assessment of plant effect is possible from these data and they were not investigated further.
TABLE 45. COPPER CONTENT IN WATER SAMPLES
(mg/fi)
c,. .. Period
aiauuii
1
2
3
4
5
6
7
8
9
10
MeanT
Std Dev
1
0.0070
0.0073
N.D.*
0.0078
0.0175
N.D.
0.0084
0.0136
N.D.
0.0198
0.0076
0.0085
0.0062
2
0.0228
0.0046
0.0400
0.0188
0.0072
0.0101
0.0746
0.0126
0.0461
0.0065
0.0243
0.0227
*N.D.-none detected. Detection limit: 0.0032 mg/S.
tSummary statistics calculated using 0.0016 for N.D. values.
117
-------
Cadmium-Jhe results of these analyses for cadmium content in the water samples are shown in Table 46.
As can be seen, the vast majority of these determinations fell at or below the detection limit of the method.
Only three values were above 0.0007 mg/Jl, and none of these occurred in the two post-operational periods.
The high frequency of results below the detection limit makes a formal statistical analysis of the data
unwarranted. Since the results of the Periods 4 and 5 analyses were low, however, it can be concluded that
there was no significant effect on the water due to cadmium arising from the plant operation.
TABLE 46. CADMIUM CONTENT IN WATER SAMPLES
(mg/fi)
Period
aiauon
1
2
3
4
5
6
7
8
9
10
Meant
Std Dev
1
0.0008
0.0013
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
-
-
2
0.0021
N.D.
0.0007
N.D.
N.D.
N.D.
0.0022
N.D.
N.D.
N.D.
0.00074
0.00075
4
N.D.*
N.D.
N.D.
N.D.
N.D.
N.D.
0.0007
N.D.
N.D.
N.D.
0.00037
0.00011
5
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
-
-
*N.D.-none detected. Detection Limit: 0.00067 mg/fi.
tSummary statistics calculated using 0.00034 for N.D. values.
Mercury-The analysis of the water samples for mercury content produced the results shown in Table 47.
One-half of the detection limit was substituted for the low values and the data submitted to an ANOVA pro-
cedure. The ANOVA is summarized in Table 48.
The sums of squares and mean squares shown in the table were obtained from using a factor of 10s times
the observed concentration. This is done for ease of computation and presentation.
The periods factor is significant beyond the one-percent level and the two sub-hypotheses of interest are
tested. The pre- vs. post-operational contrast is significant beyond the one-percent level. The values of the
period means thus indicate that there was an increase in the observed levels of mercury after the plant went
into operation. The magnitude of the difference, however, is only 2.2 x 10"5, and may not be large enough to
be of practical significance.
TABLE 47. MERCURY CONTENT OF WATER SAMPLES
(mg/fi)
_ . Period
Station
1
2
3
4
5
6
7
8
9
10
*N.D.-none detected. Detection limit: 0.00002 mg/fi.
1
0.00004
0.00004
0.00003
0.00002
N.D.*
0.00004
0.00003
0.00002
0.00002
N.D.
2
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
0.00002
N.D.
4
0.00003
N.D.
0.00005
0.00005
0.00005
0.00009
N.D.
0.00008
0.00004
0.00003
5
0.00003
0.00003
0.00003
0.00005
0.00008
0.00005
0.00005
0.00003
N.D.
N.D.
118
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TABLE 48. ANOVA ON MERCURY IN WATER SAMPLES
(mg/mix 10-5)
Source
Periods
I,2vs4,5
4vs5
Sites
Period X Sites
(error)
Total
df
3
1
1
9
27
39
SS
62.10
48.40
1.23
30.40
85.40
177.90
MS
20.70
48.40
1.23
3.38
3.16
F
6.55
15.32
<1
Significance
Level
<0.01
<0.01
N.S.
Period Means (mg/ml X 10~*)*'
1 2 45
2.60 1.10 4.40 3.70
The sites factor was not significant for these data and the conclusion is that the change in the overall
levels of mercury in water was unrelated to distance from the plant. For mercury in water samples, then, there
is no evidence that the changes in the environmental levels are related to the treatment plant operation.
Zr/7C-Analyses for concentration of zinc in the water samples were made only on samples from the first
two sampling periods. The mean for Period 1 was 0.0512 mg/i, and for Period 2 was 0.0549 mg/4. The
analytical results and the means and standard deviations are shown in Table 49.
Since only pre-operational data are available, there was no assessment of the effect of the plant on the
surrounding area possible from these data.
TABLE 49. ZINC CONTENT OF WATER SAMPLES
(mg/fi)
Ct ,. Period
Station
1
2
3
4
5
6
7
8
9
10
Meant
Std Dev
*N.D.-none detected. Detection limit: 0.0223 mg/B.
tSummary statistics calculated using 0.0112 in place of N.D. values.
Findings-
The findings of the study related to environmental water samples are summarized in Table 50. For the
microbiological analyses, total coliform results showed a tendency to increase from the pre- to the post-
operational period, but no relationship of the distance to the plant to these increases could be found. The
post-operational standard plate count data showed a decrease from the earlier levels, but this is attributable to
the Period 1 results and is considered questionable. The remaining three periods were not significantly dif-
ferent from one another. Thus, no relationship to the sewage treatment plant is indicated by these data.
1
0.0581
0.0653
N.D.*
0.0407
0.0546
0.1499
0.0476
0.0290
0.0244
0.0709
N.D.
0.0512
0.0386
2
0.0566
N.D.
0.1305
0.0443
N.D.
N.D.
0.2020
0.0591
N.D.
N.D.
0.0549
0.0642
119
-------
TABLE 50. STATISTICAL FINDINGS SUMMARY-WATER SAMPLES
Statistical Inferences Indicated Relation-
Seasonal Effect Operational Effect Distance Effect ship to Sewage
(Winter vs Fall) (Baseline vs Operational) (Near vs Far) Treatment Plant
Microbiological
Total Coliform
Standard Plate Count
Fall increase Operational increase No difference
No difference Operational decrease (?) -
None
None
Trace Metals
Cadmium, copper, zinc
Lead
Mercury
No statistical analysis warranted
No difference No difference -
No difference Operational increase No difference
None
None
The cadmium, copper, and zinc results did not permit an assessment of the treatment plant effect. The
lead data showed no period effects and no relationship between sites due to distance from the plant. The
operational levels of mercury were higher than the baseline levels, but no relationship to nearness to the plant
could be detected. The trace metal data, then, as did the microbiological, give no indication of a relationship
between the operation of the sewage treatment plant and the trace metal content of the environmental water
samples.
HOUSEHOLD HEALTH SURVEY
Household and Personal. Characteristics
The information obtained on the households and the residents therein was obtained for two purposes;
first, to describe the populations sampled, and second, to allow a comparison to be made between the two sets
of households. To fully assess the health information obtained and to evaluate the results, the two samples
should be well-matched or the differences must be known and accounted for in the conclusions drawn. The
questionnaire data relating to the household itself, the heads of household, and the individuals in residence
are summarized below for both surveys and compared for consistency.
Household Distributions-
The number of households surveys was 1107 in 1974 and 1109 in 1976. Information on households was
collected in two general categories; socioeconomic characteristics and exposure characteristics.
Socioeconomic status was evaluated by the education and occupation of the head of the households, the size
of the household, the race of the individual interviewed, and the number of bedrooms in the house. The
number of bedrooms is presumed to indicate the economic status of the household to a limited degree.
Exposure characteristics included the location, with respect to the map sectors discussed above, distance from
the Egan plant, distance from other potential sources of pollutants such as roadways or industries, length of
residence at that address, length of residence in that city or town, and extent of air-conditioning. The
information obtained in the survey is summarized below for those categories.
Socioeconomic Factors-The education of the heads of the households was obtained and is summarized in
Table 51. Of the 1100 heads of household in 1974, 63 percent had some college and 35 percent had completed
college. The 1976 data were similar, with percentages of 64 and 35, respectively. Compared to the 1970 census
data, only 7.2 percent of those in the surrounding area had completed 4 years of college, but the changes in
the resident's characteristics in the period between 1970 and 1976 discussed in Section 4 above accounts for
much of this difference.
120
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TABLE 51. COMPARATIVE EDUCATIONAL LEVELS
Educational Level of
Head of Household Baseline Operational
8th Grade
High School Incomplete
High School Complete
College Incomplete
College Complete
Graduate School
29
38
340
309
277
107
30
48
339
311
265
113
The occupations of the heads of households are shown in Table 52 grouped into general classifications.
As can be seen, there was a shift in the occupational distribution between the 1974 and 1976 surveys. The
percentage of people employed in professional and administrative positions was 17.3% in 1974 and 34.0% in
1970. The 1970 census data showed 31.3% of the total employed being in professional or managerial posi-
tions. The shift in the occupations of the persons surveyed also probably reflects the shift in the residential
nature of the area discussed above.
TABLE 52. COMPARATIVE OCCUPATIONAL DISTRIBUTIONS
Occupation of
Head of Household
Professional
Administrative
Technical
Clerical
Sales
Foremen/Craftsmen
Laborer
Teacher
Public Assistance
Retired
Baseline
73
118
192
68
150
274
105
25
20
32
Operational
112
194
178
24
158
196
74
24
26
46
The distribution of the size of the residences involved in the survey is shown in Table 53. The number of
bedrooms as an indicator of relative affluence is imperfect, but the frequency of three- and four-bedroom
dwellings gives an indication that the exposed population is above the lower class or poverty-level income.
The increase in relative frequency of smaller bedroom dwellings probably results from apartment complexes
included in the 1976 survey which did not exist at the time of the 1974 survey. The majority of apartments are
two-bedroom, so the results in the table should not be interpreted to reflect a shift downward in the income
level over the two-year span.
TABLE 53. COMPARATIVE SIZE OF RESIDENCES
Number of Bedrooms Baseline Operational
1
2
3
4
5 +
4
110
693
257
43
16
191
618
245
37
121
-------
The households were identified by sector on the survey, with the sector designation being made according
to the map shown in Figure 2 in Section 4. The sectors and the numbers of households surveyed in each are
summarized in Table 54. The baseline survey of 61 Lexington Green Apartment households within 700 m of
the plant site was actually made in late September 1974. In the baseline survey, the pattern is somewhat
irregular, but at least 19 households were obtained in each sector and over 50 in the majority. In the opera-
tional survey, the workers obtained a nominal 50 from each sector, with one or two extras in five of the sec-
tors. As such, they do not necessarily reflect the population density in those sectors, but rather a sampling of
the households from various distance-direction locations.
TABLE 54. DISTRIBUTION OF HOUSEHOLDS BY SECTOR
Area of Residence by Sector Baseline Operational
Lexington Green Apartments
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
61
33
44
67
65
50
57
56
47
51
51
51
25
51
49
19
52
51
50
75
51
51
50
51
50
50
50
51
50
50
51
50
52
50
50
50
51
50
52
50
50
51
50
50
The households were classified by the number of members in the family, and this is summarized in Table
55. As can be seen, there is a wide range of sizes, with a significant percentage for both years falling in the
groups with between two and six members, inclusive. This range spans the expected family size of an
American family and is considered to be representative.
TABLE 55. COMPARATIVE FAMILY SIZE DISTRIBUTIONS
Number of Members Baseline Operational
1
2
3
4
5
6
7
8
9+
24
189
188
324
226
94
36
14
12
34
192
227
317
206
77
36
14
6
122
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The racial characteristics of the surveyed individuals conform to those of the area, with the great majori-
ty of the persons interviewed identified by the interviewers as white. The summary of the findings is shown in
Table 56. In 1974, the interviewer failed to record the race of the subject in 153 of the cases, but the percen-
tage of whites among the responses was 96.7. In 1976, the percentage of whites was 96.5, which corresponds
well. The 1970 census of the surrounding area showed 99.3% classified as white, so these figures may be said
to be representative of the area.
TABLE 56. COMPARATIVE RACIAL DISTRIBUTIONS
Baseline Operational
Race
None indicated
Indian
Mexican-American
Negro
Oriental
White
Other
TOTAL
No.
153
2
6
8
5
860
8
1042
%
14.68
0.19
0.58
0.77
0.48
82.53
0.77
No.
0
7
11
9
12
1069
0
1108
%
0.0
0.63
0.99
0.81
1.08
96.48
0.0
Exposure Characteristics-^^ distance of each household from the Egan plant was recorded. For the
baseline survey, the distances were determined for one-kilometer circles around the plant, while the opera-
tional survey used one-half-kilometer circles. For comparison of the two groups, then, frequency classes of
less than 1.0 km, 1.0 to 2.0, 2.0 to 3.0, 3.0 to 4.0, and 4.0 to 5.0 kilometers from the plant were used. The
household distance distribution is shown in Table 57.
TABLE 57. COMPARATIVE DISTANCES FROM TREATMENT PLANT
Distance of Residence
from Egan Plant, km Baseline Operational
0.0-1.0
1.0-2.0
2.0-3.0
3.0-4.0
4.0-5.0
67
167
194
319
360
82
94
238
333
358
To indicate exposure to sources other than the sewage treatment plant, the distance of the residences
from expressways, major highways, and industries were recorded. Roadways are a source of trace metal air
pollution, particularly lead and cadmium. Industries are a source of metals and other air pollutants which can
affect the health of the populace. The summary tables for these criteria of classification are presented in
Tables 58 through 60.
In 1976, the households obtained tended to be closer to expressways than in 1974. This was primarily in
the 2 blocks to 1 mile range, where the percentage changed from 27.4 to 39.0. A similar pattern is seen for the
relative distance to major highways. For this category, however, there was also a higher frequency in the
close-in category. The percentages in these two exposure categories increased from 17.2 to 28.3, and from
33.1 to 36.8, respectively, for the two relatively high exposure categories.
The distance to an industrial source followed an irregular pattern. More households in the operational
survey fell in the close-in category, but fewer fell in the intermediate classification. Overall, however, the ma-
jority of the households were more than one mile from any such source of pollutants in both periods.
123
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TABLE 58. COMPARATIVE DISTANCES FROM NEAREST EXPRESSWAY
Distance Baseline Operational
Less than 2 blocks 56 71
2 blocks up to 1 mile 303 433
At least 1 mile 748 605
TABLE 59. COMPARATIVE DISTANCES FROM NEAREST
MULTILANE HIGHWAY
Distance Baseline Operational
Less than 2 blocks
2 blocks up to 1 mile
1 mile or more
190
366
551
314
519
276
TABLE 60. COMPARATIVE DISTANCES FROM
NEAREST INDUSTRY
Distance Baseline Operational
Less than 2 blocks
2 blocks up to 1 mile
1 mile or more
25
135
947
46
64
999
The length of residency of the individuals was recorded, both at the address and in the city in which the
household was located. These were classified into less than one, one to three, and three or more years for pur-
poses of summarizing the results. The residency at present address summary is shown in Table 61, and
residency in city in Table 62.
TABLE 61. COMPARATIVE YEARS OF RESIDENCY
AT PRESENT ADDRESS
Years Baseline Operational
Less than 1 259 220
1-3 296 282
3 or more 552 607
TABLE 62. COMPARATIVE YEARS OF RESIDENCY IN THE CITY
Years Baseline Operational
Less than 1
1-3
3 or more
222
271
614
178
249
682
The distributions for the two surveys are similar for the length of residency at the address. Of interest
also is the fact that of the households in the operational survey, 80.2 percent had been at that address for one
year or more, spanning the period during which the plant had been operational. The residency in the city
distribution was similar, with 83.9 percent being in the city for at least one year, and exposed to the plant's en-
tire period of operation. It is also noteworthy that 67.9 percent of the 1976 households had resided at least
two years in the area, so they could also have been surveyed in 1974.
124
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The final exposure characteristic concerns the extent of air-conditioning in the home. It has been noted
that air-conditioning in the home removes many of the particles from the air which carry trace metals or other
contaminants. The distribution of the households with regard to air-conditioning is shown in Table 63. The
residences are divided into categories of none, window units, and central air-conditioning.
TABLE 63. COMPARATIVE HOUSEHOLD AIR
CONDITIONING DISTRIBUTIONS
Air Conditioning Baseline Operational
None 277 195
Window 224 242
Central 604 670
The proportion with no air-conditioning decreased in 1976 from 25.0 percent to 17.6 percent. The pro-
portion of window units was comparable in both surveys, while the increase occurred in the central system
category. The developing nature of the area accounts for some of this, since most new homes are designed
with a central unit, as does the increase in apartment houses following the baseline survey.
Household Member Distributions-
Distribution of Age and Sex-The households surveys in 1974 had 4428 members, while there were 4269
members in the 1976 households. The members of the households surveyed in both 1974 and 1976 were
classified by age and are summarized in Table 64 for each sex. Fifteen age groupings were used for this com-
parison: 0-4, 5-9, 10-14, 15-19,...65-69, and 70 and over.
The operational survey contained a lower frequency of children up to 14 years. In both surveys, the
higher frequencies were those in the children and young adult classes spanning the ages from 0 to 39 in both
sexes, with a slight decrease from the ages 20 to 24. This is consistent with the results cited above in the
household characteristics and the growth of apartment dwellings in the interim between the two surveys.
Distribution by Age and Distance-To investigate the possibility that the observed differences in the
incidence of illnesses, diseases, and symptoms in the household health survey resulted from inconsistencies
between the two samples taken in 1974 and in 1976, the individuals were classified by their age group and
relative to the distance from their residence to the Egan plant.
TABLE 64. COMPARATIVE AGE AND SEX DISTRIBUTIONS
Age
Baseline
Operational
Male
0-4
5-9
10-14
15-19
20-24
25-29
30-34
35-39
40-44
45-49
50-54
55-59
60-64
65-69
70+
279
347
276
202
87
153
238
200
138
115
94
72
35
17
22
224
271
250
251
. 127
162
224
154
152
118
107
59
25
16
29
(continued)
125
-------
TABLE 64. (continued)
Age
Baseline
Operational
Female
0-4
5-9
10-14
15-19
20-24
25-29
30-34
35-39
40-44
45-49
50-54
55-59
60-64
65-69
70+
250
298
231
205
97
217
248
162
128
101
85
45
33
18
35
182
264
235
210
137
235
209
151
151
95
94
41
33
23
40
TOTALS
4428
4269
For this analysis, more general classifications of age were used which relate more closely to the frequency
of illness. The classifications are: 0-5, 6-18, 19-45, 46-64, and 65 For each category, the total is subdivided by
distance from the plant in three classes: 0-2.0 km, 2.0-3.5 km, and 3.5 to 5.0 km. These results are presented
in Table 65, along with the percentage of the total sample that lies within that combination.
TABLE 65. COMPARATIVE AGE GROUP FREQUENCIES BY DISTANCE
Baseline
Age
0-5
6-18
19-45
46-64
65 +
Distance
0-2.0
2.0-3.5
3.5-5.0
0-2.0
2.0-3.5
3.5-5.0
0-2.0
2.0-3.5
3.5-5.0
0-2.0
2.0-3.5
3.5-5.0
0-2.0
2.0-3.5
3.5-5.0
No.
146
246
288
140
580
619
358
680
746
105
169
257
17
36
41
%
3.3
5.6
6.5
3.2
13.1
14.0
8.1
15.4
16.8
2.4
3.8
5.8
0.4
0.8
0.9
Operational
No.
104
189
219
167
512
616
288
706
830
70
176
272
15
38
55
%
2.4
4.4
5.1
3.9
12.0
14.5
6.8
16.6
19.5
1.6
4.1
6.4
0.4
0.9
1.3
Difference
(Operational %
Baseline %)
-0.9
-1.2
-1.4
0.7
-1.1
0.5
-1.3
1.2
2.7
-0.8
0.3
0.6
0.0
0.1
0.4
TOTAL
4428
100
4257
100
126
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As can be seen, both sexes there was a lower frequency of individuals falling in the youngest group at all
three distances in the operational survey and a slightly higher frequency in the two older categories. Males
comprised 51.4 percent of the household members in 1974 versus 50.8 in 1976, and females were 48.6 and 49.2
percent of the total.
To indicate the representativeness of the samples, the total frequencies for each age group, irrespective of
distance, are calculated and presented in Table 66. The age groups do not match exactly, but are close enough
to indicate good agreement with respect to age between the 1970 census data in Table 3, above, and the two
samples taken. The percent of males in the 1970 population was 50.2, which also shows reasonable agreement
with these data.
TABLE 66. COMPARATIVE AGE-SEX FREQUENCY
(Percentage of Total Population)
Age
Group
0-5
6-18
19-45
46+
Baseline
8.2
15.9
19.6
7.5
Male
Operational
6.3
15.8
20.8
7.7
Baseline
7.3
14.4
21.5
6.7
Female
Operational
5.7
14.5
22.0
7.1
Shifts in Distribution Between the 1974 and 1976 Surveys-
Statist/cal Methodology-Each test for compatibility between the two survey groups was made using a chi-
square test for independence. The null hypothesis was that the baseline and operational survey distributions
were independent, so the relative frequency of any of the cross-classifications, e.g., baseline frequency of win-
dow air conditioner, can be accurately predicted by the relative frequency of the major classes. Thus no inter-
active or nonindependent causes, i.e., different distributions between years, are presumed to occur.
The model for the socioeconomic and exposure characteristics was a 2xk chi-square, with survey period
at two levels and characteristics at k levels, where k is variable depending upon the nature of the question. The
degrees of freedom for these are calculated as (2-l)(k-l) = k-1 for all cases. The test statistic is then compared
against the chi-square distribution with k-1 degrees of freedom to determine the significance of the test.
For the age-sex and age-distance comparisons, a three-way model was used to evaluate the distributions.
The degrees of freedom for the age-sex-year chi-square are computed as (15-1)(2-1)(2-1) = 14, for the 15 age
divisions, two sexes, and two years. For the distance comparisons there are (5-l)(2-l)(3-l) = 8 df for this test,
with five age groups, two years, and three distances.
The results of these chi-square tests and conclusions drawn are presented below for the variables under
study.
Socioeconomic Factors-Table 67 contains the chi-square statistics for the socioeconomic factors, along
with degrees of freedom and approximate significance levels. As can seen, the two samples were well-matched
with respect to educational level, household size, and race of interviewee, all of which had significances levels
in excess of 20 percent.
Occupational classifications and the number of bedrooms were not compatible, however, with highly
significant chi-square values. For the occupational variable, this indicates a significantly higher proportion of
the heads of household were employed in administrative and professional positions, and fewer in the crafts-
man and laborer categories.
127
-------
TABLE 67. CHI-SQUARE INDEPENDENCE TEST SUMMARY-
SOCIOECONOMIC CHARACTERISTICS
Characteristic
Education
Occupation
Size of Household
Race of Interviewee
Number of Bedrooms
df
5
9
8
4
4
X2
1.60
101.4
10.10
3.98
34.02
Significance Level
>0.20
<0.001
>0.20
>0.20
<0.001
Change in
Operational Survey
None
More professional
and administrative
classifications
None
None
Fewer bedrooms in
residences surveyed
For the number of bedrooms category, the significant chi-square value resulted from a general
shift from greater to smaller numbers of bedrooms per residence. This probably does not indicate a
decline in the affluence of the neighborhood. Instead, as noted above, the probable explanation is that
more apartment dwellings existed and were surveyed in 1976; apartments tended to have fewer
bedrooms than do houses.
Exposure Characteristics-Table 68 contains the chi-square statistics for the exposure characteristics bet-
ween the two surveys. As can be seen, all the test statistics are significant at the 5-percent level, and all but
three are significant beyond the 0.1-percent level. Thus, for these characteristics, the two survey groups did
differ.
TABLE 68. CHI-SQUARE INDEPENDENCE TEST SUMMARY-
EXPOSURE CHARACTERISTICS
Characteristic
Location
Distance From Plant
Distance to Nearest
Expressway
Distance to Nearest
Multilane Highway
Distance to Nearest
Industry
Length of Residence
at Present Address
Length of Residence
in City or Town
Air-Conditioning
df
21
4
2
2
2
2
2
2
X2 Significance Level
37.67 0.01
26.75 <0.001
39.85 <0.001
148.4 <0.001
32.93 <0.001
6.12 0.05
9.34 0.01
18.36 <0.001
Change in
Operational Survey
More balanced than
baseline
Shift from 1.0-2.0
to 2.0-3.0 km
Closer to express-
ways
Closer to highway
More within 2 blocks
Shift from less than
1 to over 3 years
Shift from less than
1 to over 3 years
More central A/C
The potential effect of these changes was investigated. First the location factor is significant due to the
balance among the sectors assured in the second survey. Fifty households were surveyed in each sector in
1976, while the number ranged from 19 to 75 in 1974. This causes the chi-square to be high, but does not
necessary imply differing exposure patterns.
128
-------
Similarly, the distance from the plant is significant, due to a higher percentage of the operational survey
lying in the 2.0-3.0 km range of the plant and fewer in the 1.0 to 2.0 km range. This could definitely affect the
outcome of the health effects study and the relative distance from the plant is thus considered in all subse-
quent analyses.
The distance from the residence to major roadways decreased for the 1976 survey in both the expressway
and multilane highway categories. The residences were closer on the whole, and this could result in higher
exposure to pollutants from automobile exhaust, including trace metals. The increased exposure to this type
of pollution is a confounding factor in the results and must be considered when drawing any conclusions.
Distance to the nearest industry was significantly but not uniformly different. Overall, the households
within one mile of the plant decreased, but the number within two blocks roughly doubled, from 25 to 46.
However, 85 to 90 percent of the households in both surveys were over one mile from the nearest industry,
and this was not considered to be a major source of confounding in the analyses.
The residency factors were similar, with both showing a decrease in the less than one year category, and
an increase in the over three years category. This does not indicate a problem, however, since more of the
residents surveyed would have been exposed during the treatment plant's operational period.
The air-conditioning of residences was more extensive in the households surveys in the operational
period. Air-conditioning eliminates particles from the air which might contain sources of disease and health
hazard. Thus, if this difference affects the disease and symptom results of the health survey, it would be ex-
pected to lower the occurrence of the diseases and symptoms under study during the 1976 operational period
survey.
Age-Sex Distribution-The chi-squared statistic for the change in age-sex distribution of the household
members was calculated to be 123.77 with 14 df. This has a significance level of less than 0.1 percent, and thus
a significant change had occurred. Fewer children, especially in the two lowest age categories, 0 to 4 and 5 to 9
years, of both sexes were surveyed in 1976. For those symptoms or illnesses which would be expected to occur
more frequently in children, there would be an expected decrease in incidence in the operational period survey
(1976) relative to the baseline period survey (1974), all other things being equal.
Age-Distance Distribution-The chi-squared statistic for comparison of the two surveys by age group and
distance from the plant was calculated to be 134.85 and has 8 degrees of freedom associated with it. The
significance level for this test is less than 0.001, and clearly indicates that the two surveys differed in their age-
distance make-up. As in the above data, there were fewer children overall in the operational survey and fewer
at the 0-2.0 km distance.
As a result, any of the illness, diseases, or symptoms for which a potential health effect is observed must
be considered in terms of the age pattern relative to the incidence of the disease and to the distance from the
Egan plant, considered together.
Analysis of Health Survey Illness, Disease and Symptom Incidence
Statistical Methodology-
The data from the Household Health Survey are in the form of frequency counts for occurrence of a par-
ticular chronic illness, symptom, or disease. The statistical methodology used was designed to determine if
any significant changes in the frequency of occurrence were noticed between the baseline (pre-operational)
period and the operational (post-operational) period. In addition, any increase in incidence must be related to
the distance of the household to the Egan plant, if the sewage treatment plant is to be implicated as a potential
health hazard.
129
-------
A multiple approach was used to evaluate the health data. First, a contingency table was constructed with
factors of survey period at two levels (baseline and operational) and distance at three levels (0-2.0 km, 2.0-3.5
km, and 3.5-5.0 km). A chi-square test of independence was then used to determine if these factors were
related in some fashion. The null hypothesis is that incidence bears the same relationship to distance in both
years. Rejection of the null hypothesis indicates that the pattern of occurrence of a particular illness or symp-
tom may be dependent upon the survey period and distance from the plant where the data was taken. The test
statistic is compared against a chi-square distribution with (2-l)(3-l) = 2 df to determine significance.
There can be a problem in interpreting the results of the chi-square test if the number of individuals in
each category (i.e., survey period-distance combination) varies. For this reason, the chi-square test is only us-
ed as a first indication of potential changes.
To further evaluate the test results, the relative frequency of occurrence of each symptom is used in a pre-
vs post-comparison. The test statistic for this comparison is a standard normal deviate, calculated to test the
hypothesis of equivalent proportions in the two surveys in the same distance range. The test statistic is
calculated according to the formula
where
Pi - relative frequency of occurrence in period 1,
p2 - relative frequency of occurrence in period 2,
n, - number of individuals in period 1,
n2 - number of individuals in period 2, and
p = (n^i + n2p2)/(nj + n2), relative frequency under the null hypothesis
The same test was used for the relative frequencies for the three distances separately to determine if any
change in incidence had a distance pattern consistent with the hypothesis of a plant effect on community
health. Test statistics Zi, Z2, and Z3 were calculated corresponding to the three distance ranges studied: (1)
0-2.0 km; (2) 2.0-3.5 km; and (3) 3.5-5.0 km. These statistics, then, test the hypothesis of no change in the
relative frequency of occurrence of the illness, disease, or symptom at each distance and are compared to the
corresponding Z0 for consistency.
To evaluate the results of these analyses, the illnesses, diseases, or symptoms with a significant chi-square
value were first investigated. For these health parameters, the Z0 test statistic was used to indicate if an overall
trend existed in the surveyed area and if so whether it was for increased or decreased incidence. If a significant
trend existed overall, then the individual Zj were investigated to determine if there was a pattern within the
distance levels which differed from the overall trend. If the distance pattern showed a greater increase close to
the plant than at the greater distances for any parameter, it was selected for additional analysis as one which
may have been affected by the operation of the treatment plant. These patterns of increased incidence were
then further investigated to determine if other differences, such as age or sex, could have affected the results.
Since there were unequal sample sizes between the two surveys, it was possible for a significant relation-
ship to exist with distance which did not show a significant survey-distance relationship in the chi-square test.
To account for this, the data were investigated to determine if the distance pattern indicated by the Zj describ-
ed above existed for any of the remaining illnesses, diseases, or symptoms.
130
-------
The list of illnesses and symptoms on the questionnaire is fairly comprehensive. As a result, items are
included for which no relationship of the treatment plant operation to increased incidence would be expected.
The illnesses, diseases, and symptoms were evaluated by a physician prior to the data analysis and this evalua-
tion used in the interpretation of the results. Each of the 60 parameters were denoted as to whether an increase
in incidence might plausibly be related to a nearby sewage treatment plant operation. In addition, the age
group for which the highest incidence of the disease or symptom would be expected was also evaluated. The
difference in the age distributions between the two groups are then accounted for by comparing the expected
change in incidence relative to the difference in numbers surveyed of the age group most likely to have a high
incidence of the illness, disease, or symptom.
The location of the increase in incidence of any of these diseases or symptoms is also important in assess-
ing whether a treatment plant effect occurred. If increased incidence is related to the treatment plant opera-
tion, then the pattern of increased incidence should fall on the predominant directions of wind across the
plant. These were determined to be north and south of the plant, and the incidence with distance in these two
directions were investigated for those illnesses, diseases, or symptoms which have been determined by prior
analyses to be possibly indicative of a sewage treatment plant effect on the general health.
Finally, the illnesses, diseases, and symptoms exhibiting significant pattern and potential relationship to
the sewage treatment plant operation were considered together to evaluate the survey's sewage treatment
plant health implications. Related symptoms and diseases would be expected to have similar incidence
patterns.
Incidence of Illnesses, Diseases, and Symptoms-
The number and relative frequency of positive responses to the 60 chronic illnesses, diseases, and symp-
toms are shown in Table 69. The responses are categorized by distance and by survey period relative to the
number of individuals at that distance range. Shown at the top of the table is the distribution of the
individuals (household members) into the three distance categories. The total number of individuals upon
whom information was obtained was 4412 in the baseline survey and 4257 in the operational survey.
From the tables, the tendency is observed for a decrease in the percentage incidence of the majority of the
illnesses and symptoms. Exceptions to this included influenza, skin disease, diarrhea, general weakness, pain
in chest on deep breathing, skin rash: face, and vomiting, which had notably higher percenta.ge incidences in
the operational survey at the 0-2.0 km distances.
Statistical Analysis-
The statistics calculated for testing the hypothesis of no sewage treatment plant effect are shown in Table
70. Using the chi-squared statistic, the incidence of the following parameters was determined to exhibit a
significant interaction or dependence between distance and survey period at the one percent level: asthma-hay
fever; influenza; worms; and vomiting. Significant at the five percent level were: dysentery; general weakness;
nausea; and sore throat.
As mentioned above, the illnesses and symptoms were reviewed by a physician for relationship to a treat-
ment plant and for age group expected to have the highest incidence. The summary of this review is shown as
Table 71. Using this guideline, asthma-hay fever was eliminated from further consideration as having no rela-
tionship to the operation of the treatment plant. Worms was considered a questionable disease, and for the re-
mainder, the judgment was made that changes in their incidence could result from sewage treatment plant
operation in a populated area.
To investigate these, then, the differences in their relative incidence were investigated using the standard
normal deviates, Z;, discussed above and shown in Table 70. For a possible effect to be judged, the difference
at the 0-2.0 km range should be more strongly positive than at the remainder of distances and from the overall
trend for the area under study.
131
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TABLE 69. INCIDENCE OF ILLNESSES, DISEASES, AND SYMPTOMS
REPORTED IN THE HOUSEHOLD HEALTH SURVEYS
Baseline Survey
Operational Survey
Household Members Surveyed
0.0 - 2. 0 km
Z. 0 - 3. 5 km
3. 5 - 5. 0 km
CHRONIC ILLNESSES EVER DIAGNOSED
Arthritis and Rheumatism
0. 0 - 2. 0 km
2.0 - 3. 5 km
3.5- 5. 0 km
Asthma - Hay Fever
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5- 5. 0 km
Chronic Sinusitis and Bronchitis
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5. 0 km
Diabetes
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Heart Conditions
0.0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Hypertension
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5- 5. 0 km
Kidney Disease
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Liver Disease
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Number of
Responses
N
750
1711
1951
Positive
Responses
n
33
69
91
102
188
176
78
131
175
9
23
24
8
31
27
27
48
69
6
12
16
3
5
4
Percentage
Incidence
p
4.
4.
4.
13.
11.
9.
10.
7.
9.
1.
1.
1.
1.
1.
1.
3.
2.
3.
0.
0.
0.
0.
0.
0.
4%
0%
7%
6%
0%
0%
.4%
7%
0%
2%
3%
2%
1%
8%
4%
6%
8%
5%
8%
7%
8%
4%
3%
2%
Number of
Responses
N
644
1621
1992
Positive
Responses
n
25
57
70
72
182
235
47
139
145
4
19
28
9
28
39
19
44
82
4
10
19
5
6
2
Percentage
Incidence
P
3.9%
3. 5%
3.5% '
11.2%
11.2%
1 1 . 8% .
7.3%
8.6%
7.3%
0.6%
1. 2%
1.4%
1 . 4%
1.7%
2.0%
3.0%
2. 7%
4. 1%
0.6%
0.6%
1.0%
0.8%
. 0.4%
0. 1%
(continued)
132
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TABLE 69. (continued)
Baseline Survey
Operational Survey
Malignancies
0.0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Rheumatic Heart Disease
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Thyroid Disease
0. 0 - 2.0 km
2. 0 0 3. 5 km
3. 5 - 5. 0 km
Tuberculosis
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
DISEASES DIAGNOSED DURING PAST YEAR
Anemia
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5.0 km
Croup
0. 0 - 2.0 km
2. 0 - 3. 5 km
3.5 - 5.0 km
Dysentery
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5.0 km
Empyema
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5.0 km
Infectious Jaundice
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5.0 km
Influenza
0. 0 - 2. 0 km
2.0 - 3. 5 km
3.5 - 5. 0 km
Positive
Responses
n
8
3
14
7
8
14
12
38
17
2
8
2
12
19
24
14
28
21
2
22
13
0
2
3
1
3
1
70
221
366
Percentage
Incidence
p
1. 1%
0.2%
0.8%
0. 9%
0.5%
0.7%
1.6%
2.2%
0.9%
0.3%
0. 5%
0. 1%
1.6%
1. 1%
0.7%
1.9%
1.6%
1.1%
0.3%
1.3%
0.7%
0.0%
0. 1%
0.2%
0.1%
0.2%
0. 1%
9.3%
12. 9%
18. 8%
Positive
Respons'es
n.
2
5
11
5
10
12
9
22
18
3
3
3
5
11
21
10
13
24
3
1
7
0
1
0
0
1
2
72
122
182
Percentage
Incidence
P
0.
0.
0.
0.
0.
0.
1.
1.
0.
0.
0.
0.
0.
0.
1.
. 1.
0.
1.
0.
0.
0.
0.
0.
0.
0.
0.
0.
11.
7.
9.
3%
3%
6%
8%
6%
6%
4%
4%
9%
5%
2%
2%
8%
7%
1%
6%
8%
2%
5%
1%
4%
0%
1%
0%
0%
1%
1%
2%
6%
1%
(continued)
133
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TABLE 69. (continued)
Baseline Survey
Operational Survey
Positive
Responses
n
Pleurisy
0.0- 2. 0 km
2.0 - 3. 5 km
3. 5 - 5. 0 km
Pneumonia
0. 0 - 2.0 km
2. 0-3.5 km
3.5- 5.0 km
Polio
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Skin Disease
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Sleeping Sicknesses
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5.0 km
Spinal Meningitis
0.0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Worms
0.0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
SYMPTOMS EXPERIENCED IN PAST 3 MONTHS
Bloody Diarrhea
0. 0 - 2. 0 km
2.0 - 3.5 km
3.5- 5.0 km
Bloody Urine
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5. 0 km
2
11
8
10
28
28
0
2
0
4
28
27
0
3
1
0
2
3
7
27
11
0
2
2
3
1
3
Percentage
Incidence
P
0.3%
0.6%
0.4%
1.3%
1.6%
1.4%
0. 0%
0. 1%
0. 0%
0. 5%
1.6%
1 . 4%
0. 0%
0. 2%
0. 1%
0.0%
0. 1%
0.2%
0.9%
1.6%
0. 6%
0.0%
0. 1%
0. 1%
0. 4%
0. 1%
0. 2%
Positive
Responses
n
2
5
2
5
20
31
0
0
0
11
21
27
0
0
0
0
1
5
0
0
14
2
3
5
0
4
3
Percentage
Incidence
P
0. 3%
0.3%
0. 1%
0. 8%
1.2%
1.6%
0.0%
0. 0%
'0.0%
1. 7%
1.3%
1 . 4%
0.0%
0.0%
0.0%
0.0%
0. 1%
0. 3% '
0.0%
0.0%
0.7%
0.3%
0.2%
0.3%
0.0%
0.2%
0.2%
(continued)
134
-------
TABLE 69. (continued)
Bjaseline Survey
Brown Urine
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Burning on Urination
0.0-2.0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Canker Sores Around Mouth
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5- 5. 0 km
Cold
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 -'5. 0 km
Colicky Pains in Abdomen
0.0-2.0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Convulsions
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Cough
0. 0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Cough up Blood
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Diarrhea
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5. 0 km
Draining Ear
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km .
Positive
Responses
n
0
0
2
6
14
14
21
39
71
238
451
493
9
8
17
0
2
2
143
314
293
0
0
1
31
73
•94
8
8
20
Percentage
Incidence
P
0. 0%
0.0%
0. 1%
0.8%
0.8%
0. 7%
2.8%
2. 3%
3.6%
31. 7%
26. 4%
25. 3%
1 . 2%
0. 5%
0. 9%
0. 0%
0. 1%
0. 1%
1.9. 1%
18. 4%
15. 0%
.0. 0%
0. 0%
0. 1%
4. 1%
4. 3%
4. 8%
1. 1%
0. 5%
1. 0%
Positive
Responses
n
0
0
0
6
8
16
26
38
49
182
395
474
13
1 1
13
0
0
1
120
228
268
2
2
3
49
77
85
4
17
19
Percentage
Incidence
p
0.0%
0.0%
0. 0%
0. 9%
0. 5%
0. 8%
4. 0%
2.3%
2. 5%
28. 3%
24. 4%
23. 8%
2.0%
0. 7%
0. 7%
0.0%
0.0%
0. 1%
18.6%
14. 1%
13. 5%
0.3%
0. 1%
0.2%
7.6%
4.8%
4. 3%
0.6%
1. 0%
1.0%
(continued)
135
-------
TABLE 69. (continued)
Baseline Survey
Operational Survey
Fever above 103°F
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5- 5.0 km
General Weakness
0. 0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Hemorrhafiic Rash
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5.0 km
Nausea
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Pain in Chest on Deep Breathing
0. 0 - 2.0. km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Severe Dizziness
0.0 - 2.0 km
2.0 - 3. 5 km
3. 5 - 5. 0 km
Severe Headache not Relieved by Aspirin
0. 0 - 2.0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Severe Night Sweats
0.0 - 2.0 km
2.0 - 3.5 km
3.5 - 5. 0 km
Severe Pain in Bones and Joints with High Fever
0.0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5.0 km
Severe Trouble with Teeth
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5.0 km
Positive
Responses
n
34
33
59
5
17
29
0
0
0
9
41
59
4
27
28
7
7
21
34
71
99
5
10
8
8
9
15
4
14
26
Percentage
Incidence
P
4.5%
1 . 9%
3. 0%
0. 7%
1.0%
1.5%
0. 0%
0.0%
0. 0%
1.2%
2. 4%
3.0%
0. 5%
1.6%
1. 4%
0. 9%
0. 4%
1. 1%
4. 5%
4. 1%
5. 1%
0.7%
0. 6%
0.4%
1.1%
0.5%
0.8%
0. 5%
0.8%
1.3%
Positive
Responses
n
13
29
24
12
7
11
0
2
2
19
23
33
12
24
22
3
14
14
24
65
72
4
8
9
6
16
19
5
14
20
Percentage
Incidence
P
2. 0%
1 . 8%
1 . 2%
1 . 9%
0.4%
0.6%
0. 0%
0. 1%
0. 1%
3.0%
1 . 4%
1.7%
1.9%
1. 5%
1.1%
0.5%
0. 9%
0. 7%
3.7%
4. 0%
3.6%
0.6%
0.5%
0. 5%
0. 9%
1. 0%
1.0%
0. 8%
0.9%
1.0%
(continued)
136
-------
TABLE 69. (continued)
Baseline Survey
Operational Survey
Severe Weight Loss
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Shortness of Breath
0. 0 - 2. 0 km
Z. 0 - 3. 5 km
3.5- 5. 0 km
Skin. Rash: Arms and Leus
0. 0 - 2. 0 km
2. 0 - 3.5 km
3.5- 5. 0 krn
Skin Rash: Body
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Skin Rash: Face
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Sore Throat
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Stiff Neck with Fever
0.0- 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Stiff Neck with Rash
0.0 - 2. 0 km
2. 0 - 3. 5 km
3. 5 - 5. 0 km
Uncouscious (Not Due to Blow on Head)
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5.0 km
Vomiting
0. 0 - 2. 0 km
2.0 - 3. 5 km
3. 5 - 5. 0 km
Positive
Re sponses
n
3
2
6
10
16
20
10
46
52
11
30
49
3
37
50
132
269
333
4
4
5
1
0
7
0
4
2
10
52
53
Percentage
Incidence
P
0. 4%
0. 1%
0.3%
1.3%
0. 9%
1.0%
1.3%
2. 7%
2. 7%
1 . 5%
1. 8%
2. 5%
0.4%
2. 2%
2. 7%
17.6%
1 5. 7%
17. 1%
0. 5%
0. 2%
0.3%
0. 1%
0. 0%
0. 4%
0. 0%
0.2%
0. 1%
1.3%
3.0%
2.7%
Positive
Responses
n
0
2
3
3
17
18
8
25
42
12
31
33
7
25
30
59
195
243
5
4
12
0
0
4
0
0
0
20
23
47
Percentage
Incidence
P
0. 0%
0. 1%
0. 2%
0.5%
1 . 0%
0. 9%
1. 2%
1. 5%
2. 1%
1.9%
1 . 9%
1.7%
1.1%
1. 5%
1. 5%
9. 2%
12.0%
12. 2%
0.8%
0. 2%
0.6%
0.0%
0. 0%
0. 2%
0. 0%
0.0%
0. 0%
3.1%
1.4%
2.4%
(continued)
137
-------
TABLE 69. (continued)
Baseline Survey
Weakness of Arms or Legs
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5- 5. 0 km
Positive
Responses
n
10
10
22
Pe rcentage
Incidence
1.3%
0. 6%
1.1%
Operational Survey
Positive Percentage
Responses Incidence
n P
4
10
11
0.6%
0. 6%
0.6%
Yellow Skin
0. 0 - 2. 0 km
2.0 - 3. 5 km
3.5- 5. 0 km
Yellow Eyeballs
0. 0 - 2. 0 km
2. 0 - 3. 5 km
3.5 - 5. 0 km
0.0%
0. 0%
0. 0%
0. 1%
0.0%
0. 1%
0.0%
0.0%
0.0%
0.2%
0. 1%
0. 4%
138
-------
TABLE 70. TEST STATISTICS FOR SIGNIFICANCE OF RESPONSES
Illness, Disease or Symptom
Arthritis and Rheumatism
Asthma-Hay Fever
Chronic Sinusitis and Bronchitis
Diabetes
Heart Conditions
Hypertension
Kidney Disease
Liver Disease
Malignancies
Rheumatic Heart Disease
Thyroid Disease
Tuberculosis
Anemia
Croup
Dysentery
Emphysema
Infectious Jaundice
Influenza
Pleurisy
Pneumonia
Polio
Skin Disease
Sleeping Sickness
Spinal Meningitis
Worms
Chi-Square Z0
Chronic Illnesses Ever Diagnosed
0.11 -1.88
13.19f 1.43
6.85 -1.52
2.38 . -0.28
1.70 1.08
2.68 0.40
0.82 0.04
1.22 0.30
3.41 -0.94
0.64 -0.12
1.98 -1.47
2.29 . -0.57
Diseases Diagnosed During Past Year
1.76 -1.70
4.11 -1.33
10.03* -3.63
1.20 -1.59
1.96 -0.65
15.06t -8.64
1.25 -2.09
2.35 -0.69
0.00 -1.39
4.27 0.22
0.00 -1.96*
0.75 0.37
24.96t -3.90f
z,
-0.48
-1.36
-2.02*
-1.12
0.56
-0.68
-0.39
0.93
-1.67
-0.32
-0.31
0.62
-1.40
-0.45
0.62
0.00
-0.93
1.14
0.15
-1.00
0.00
2.12*
0.00
0.00
-2.46|
Z2
-0.78
0.22
0.97
-0.45
-0.18
-0.16
-0.30
0.39
. 0.78
0.59
-1.87
-1.42
-1.32
-2.18
-4.27
-0.53
-0.95
-5.12
-1.40
-0.98
-1.38
-0.82
-1.69
-0.53
-5.08|
Z3
-1.82
2.85t
-1.94*
0.48
1.40
0.95
0.45
-0.84
-0.65
-0.45
0.11
0.42
-0.57
0.38
-1.39
-1.75
0.56
-8.73
-1.93
0.31
0.00
-0.08
-1.01
0.68
0.55
Symptoms Experienced In Past 3 .Months
Bloody Diarrhea
Bloody Urine
Brown Urine
Burning on Urination
Canker Sores Around Mouth
Cold
Colicky Pains in Abdomen
Convulsions
Cough
Cough Up Blood
Diarrhea
Draining Ear
Fever Above 103; F
General Weakness
Hemorrhagic Rash
Nausea
Pain Chest on Deep Breathing
Severe Dizziness
Severe Headache Not Relieved
by Aspirin
*Significant at 0.05 level. Required: X2
•^Significant at 0.01 level. Required: X2
1.12 1.68
4.80 0.07
0.00 -1.39
1.53 -0.34
3.27 -0.86
3.88 -2.14*
1.61 0.52
0.83 -1.30
3.66 -3.16f
1.14 2.18
4.20 1.07
4.40 0.63
6.26 -4. lOt
10.40* -2.17*
0.00 2.04*
10.04* -2.26*
4.89 0.12
5.11 -0.33
1.20 -1.92
- 7. 84, Z- 1.96.
- 11. 34, Z- 2.58.
1..52
-1.61
0.00
0.27
1.28
-1.41
1.22
0.00
-0.21
1.53
2.78*
-0.90
-2.59t
2.03*
0.00
2.32*
2.32*
-1.03
-0.75
0.51
1.40
0.00
-1.17
0.12
-1.32
0.81
-1.38
-3.35f
1.45
0.67
1.94
-0.30
-1.92
1.45
-2.05*
-0.23
1.66
-0.20
l.ll
-0.03
-1.43
0.31
-2.16*
-1.08
-0.79
-0.60
-1.41
0.98
-0.83
-0.23
-3.98|
-2.93f
1.40
-2.84|
-0.93
-1.25
-0.25
(continued)
139
-------
TABLE 70. (continued)
Illness, Disease or Symptom
Chi-Square
Symptoms Experienced in Past 3 Months (Cont'd)
Severe Night Sweats
Severe Pain in Bones and Joints
with High Fever
Severe Trouble with Teeth
Severe Weight Loss
Shortness of Breath
Skin Rash: Arms and Legs
Skin Rash: Body
Skin Rash: Face
Sore Throat
Stiff Neck with Fever
Stiff Neck with Rash
Unconsciousness (Not Due to
Blow on Head)
Vomiting
Weakness of Arms or Legs
Yellow Skin
Yellow Eyeballs
0.30
1.63
0.59
2.04
3.17
1.60
2.02
3.90
8.45*
1.18
0.55
0.00
12.04f
2.06
0.00
1.76
-0.17
1.23
-0.37
-1.42
-0.70
-2.20*
-0.84
-2.05*
-6.55f
1.49
-1.09
-2.40
-1.48
-1.92
0.00
2.18
-0.11
-0.25
0.56
-1.61
-1.68
-0.15
0.58
1.52
-4.57f
0.56
-0.93
0.00
2.27*
-1.33
0.00
0.11
-0.36
1.54
0.14
0.05
0.33
-2.29
0.34
-1.32
-3.08f
0.08
0.00
-1.95
-3.15f
0.12
0.00
1.03
*Significant at 0.05 level. Required: XJ - 7.84, Z -1.96.
tSignificant at 0.01 level. Required: X1 - 11.34, Z - 2.58.
TABLE 71. PHYSICIAN'S EVALUATION OF RESPONSE CATEGORIES
Illness, Disease, or Symptom
Expected Sewage
Treatment Plant Effect
For Expected Effects,
Age Group
with Highest Incidence
0.20
0.63
-0.96
-1.03
-0.39
-1.15
-1.88
-2.35
-4.33f
1.66
-0.94
-1.43
-0.71
-1.98
0.00
2.09*
Chronic Illnesses Ever Diagnosed
Arthritis and Rheumatism
Asthma-Hay Fever
Chronic Sinusitis and Bronchitis
Diabetes
Heart Conditions (Too Broad)
Hypertension
Kidney Disease
Liver Disease
Malignancies
Rheumatic Heart Disease
Thyroid Disease
Tuberculosis
No
No
Yes
No
No
No
Yes
Yes
No
Yes
No
Yes
—
-
None
_
-
-
20-50
2040
-
Below 20
-
None
(continued)
140
-------
TABLE 71. (continued)
Illness, Disease, or Symptom
Anemia
Croup
Dysentery
Emphysema
Infectious Jaundice
Influenza
Pleurisy
Pneumonia
Polio
Skin Disease
Sleeping Sickness
Spinal Meningitis
Worms
Bloody Diarrhea
Bloody Urine
Brown Urine
Burning on Urination
Canker Sores Around Mouth
Cold
Colicky Pains in Abdomen
Convulsions
Cough
Cough Up Blood
Diarrhea
Draining Ear
Fever Above 103° F
General Weakness
Hemorrhagic Rash
Nausea
Expected Sewage
Treatment Plant Effect
Diseases Diagnosed During Past Year
No
Yes
Yes
Possible
Yes
Yes
Possible
Yes
Yes
Possible
No
Yes
Possible
Symptoms Experienced in Past 3 Months
Yes
Possible
Yes
No
Yes
Yes
No
Possible (if related
to high fever)
Yes
Yes (infections)
Yes
Yes
Yes
Yes
Possible
Yes
For Expected Effects,
Age Group
with Highest Incidence
_
Young children
Children
-
2(MO
None
-
Infants and elderly
Infants
None
20-30
0-20
Children
None
None
None
-
None
None
-
None
None
None
Children
Children
Children
None
None
Children
Pain in Chest on Deep Breathing Possible -
Severe Dizziness
Severe Headache Not Relieved
by Aspirin
Severe Night Sweats
Severe Pain in Bones and Joints
with High Fever
Severe Trouble with Teeth
Severe Weight Loss
Shortness of Breath
Skin Rash: Arms and Legs
Skin Rash: Body
Skin Rash: Face
Sore Throat
Stiff Neck with Fever
Stiff Neck with Rash
Unconsciousness (Not Due to
Yes
No
Yes
Yes
No
Possible
No
Possible
Possible
Possible
Yes
Yes
Yes
Yes
20-30
-
20-30
None
-
None
-
None
None
None
Children
Children
Children
None
(continued)
141
-------
TABLE 71. (continued)
For Expected Effects,
Expected Sewage Age Group
Illness, Disease, or Symptom Treatment Plant Effect with Highest Incidence
Symptoms Experienced in Past 3 Months (Cont'd)
Blow on Head
Vomiting Yes Children
Weakness of Arms or Legs No -
(Possible)
Yellow Skin Yes None
Yellow Eyeballs Yes None
For influenza the nearest distance results were different from the remainder. The overall relative in-
cidence of influenza decreased significantly from the baseline to the operational period, but the decrease was
a result of the decrease at the 2.0 to 3.5 and 3.5 to 5.0 km ranges. At the 0-2.0 km distance there was an in-
crease in relative frequency of influenza, although not large enough for significance. The difference between
the two surveys was 1.9 percent (11.2-9.3), as shown in Table 69 above.
The responses for worms do not exhibit the pattern indicating a relationship to the sewage treatment
plant. There was a decrease overall and at the two closest distances, all with significance levels less than one
percent.
The responses to the symptom of vomiting show potential results of the treatment plant operation. The
overall trend was for a decrease in percentage incidence, although the calculated test statistic did not achieve
significance. There was a significant increase at the 0-2.0 km range, however, with a significance level of less
than five percent. The next distance level, 2.0 to 3.5 km, showed a significant decrease, however, and the re-
maining level had an insignificant decrease. The difference in the percentage incidence at the closest distance
was 1.8 percent (3.1-1.3).
For the dysentery responses, a decrease was noted in the overall percentage incidence as well as in the 2.0
to 3.5 km and 3.5 to 5.0 km ranges. At the 0-2.0 km distance, however, a slight increase was noted. From the
data tables, this change was from 0.3 percent to 0.5 percent, and represented an increase from 2 to 3 positive
responses.
The responses for general weakness showed a strong pattern suggestive of a sewage treatment plant in-
fluence. All the calculated statistics were significant, with the overall and the two greatest distances showing a
significant decrease in the operational period over the baseline. The 0-2.0 km data, however, showed a signifi-
cant increase between the two periods. The percentage incidence increased 1.2 percent (0.7 to 1.9) from the
baseline to the operational period.
An identical pattern was seen for the nausea responses. The test statistics for the overall area and for the
2.0 to 3.5 and 3.5 to 5.0 km ranges exceed the tabled value for the five percent significance level and indicate
decreases in relative frequency of response. The closest range, however, had a significant increase in percen-
tage incidence, going from 1.2 to 3.0 percent.
The final response, for sore throat, gave no indication of a treatment plant effect, since all four of the
calculated statistics show a decrease significant beyond the one percent level. The significance of the chi-
squared statistic in the instance represents a difference only in the relative magnitude of the decrease, with the
largest decrease seen at the 0 to 2.0 km range.
142
-------
As noted in the methodology section, the chi-squared values are first indications of a change, and may in
some instances be influenced by the differences in sample size between the two survey groups. The Z; are fur-
ther investigated to determine if a distance pattern of the type noted above exists for any of the illnesses or
symptoms which had insignificant chi-square tests. One disease and two symptoms satisfied this criterion:
skin disease, diarrhea, and pain in chest on deep breathing.
The responses for skin disease showed a significant increase at the 0 to 2.0 km range, with no change in
the relative frequency noted for the overall area or for the two greater distances. The difference in the percen-
tage incidence was from 0.5 to 1.7 percent of the responses.
A similar pattern was observed for diarrhea. The test statistic for the closest distance was significant at
the one percent level, while the 2.0 to 3.5 km data showed an insignificant increase and the 3.5 to 5.0 km data
indicated an insignificant increase in relative incidence. The change in percentage incidence was 3.5 percent
(7.6-4.1).
Finally, pain in chest on deep breathing responses showed a higher percentage incidence in the opera-
tional period, significant at the five percent level, at the 0 to 2.0 km range with decreasing frequency,
although insignificant, at the remaining distances. The percentage incidence for the individuals closest to the
plant rose from 0.5 to 1.9 percent. It should be noted, however, that the physician's evaluation of this symp-
tom indicated only a possible relationship to the operation of the sewage treatment plant.
For those symptoms for which a possible effect was indicated, the number of positive responses and the
percentage incidence was determined along the four major wind directions. Two distance levels were used for
this portion of the analysis, 0 to 3.5 km and 3.5 to 5.0 km. The sectors as shown on the area map were
classified as to principal direction from the plant and are shown in Table 72 along with the responses and
percentage incidences. The wind direction for this area as shown in Figure 12 tended to be from either the
north or the south for the period of the study, and the relative frequency in the operational period compared
to the baseline is evaluated on these two directions. However, since the wind can be from any direction over a
period of time, direction relationships cannot be given much weight.
The influenza results show a decrease at both distances for incidence on the north side, but an increase on
the south side at the closer distance. The increased percentage incidence was 3.1 percent (15.1-12.0) while for
the greater distance, there was a marked decrease from baseline to operational in the south. The data for the
north show a decrease, but it should be noted that the earlier investigation separated the 0 to 2.0 km and 2.0 to
3.5 km data. There was a marked decrease in the 2.0 to 3.5 km range in that comparison, and the data here
may be somewhat confounded because of that.
The responses for vomitting show an increase at both distances on the north side of the Egan plant and
an increase on the south side for the closer distance. The two remaining directions showed decreased percen-
tage incidence at the 0 to 3.5 km range, and the percentage incidence at the greater distance decreased mark-
edly for the south data.
No pattern was evident for dysentery for which, as noted above, the frequency of occurrence at the close
distances was small. The responses at the closer distance were balanced over the four directions.
Responses to general weakness may indicate a directional effect. There was a decrease in relative fre-
quency in all directions at the 3.5 to 5.0 km range from the baseline to the operational period. The close-in
data, however, showed an increase on both the north and south sides, of 0.4 percent and 0.5 percent,
respectively.
143
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TABLE 72. INCIDENCE BY DISTANCE AND DIRECTION FROM THE
EGAN PLANT FOR THOSE DISEASES AND SYMPTOMS WITH A
POTENTIALLY IMPLICATING DISTANCE PATTERN
Baseline Survey
Number of Included
Responses Sectors
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
0.0-3.5 km
North
East
South
West
3.5 -5 .Okm
North
East
South
West
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
0.0-3.5 km
North
East
South
West
410
476
750
825
304
513
394
756
Positive
Responses
1
17
6
0
0
6
4
3
41
89
90
71
71
84
71
141
3
14
2
13
Household Members Surveyed
1,11, Lex. Green
2,3
4,5,6
7,8,9,10
12,21
13, 14, 15
16, 17
18, 19, 20
Percentage
Incidence
Diseases Diagnosed During Past Year
Dysentery
0.2
3.6
0.8
0.0
0.0
1.2
1.0
0.4
Influenza
10.0
18.7
12.0
8.6
23.4
16.4
18.0
18.7
Skin Disease
0.7
2.9
0.3
1.6
Operational Survey
Number of
Responses
452 1,
375
595
843
366
623
361
642
Positive
Responses
1
0
1
2
4
1
1
1
32
22
90
50
32
71
22
57
6
6
10
10
Included
Sectors
11, Lex. Green
2,3
4,5,6
7,8,9, 10
12,21
13,14, 15
16,17
18, 19,20
Percentage
Incidence
0.2
0.0
0.2
0.2
1.1
0.2
0.3
0.2
7.1
5.9
15.1
5.9
8.7
11.4
6.1
8.9
1.3
1.6
1.7
1.2
(continued)
144
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TABLE 72. (continued)
3.5-5.0 km
North
East
South
West
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
Positive
Responses
4
3
3
18
16
36
25
27
11
15
27
41
1
10
4
7
2
9
3
15
7
15
13
15
5
12
12
30
Baseline Survey
Percentage
Incidence
Skin Disease (Cont 'd)
1.3
0.6
0.8
2.4
Symptoms Experienced in Past 3
Diarrhea
3.9
7.6
3.3
3.3
3.6
2.9
6.9
5.4
General Weakness
0.2
2.1
0.5
0.8
0.7
1.8
0.8
2.0
Nausea
1.7
3.2
1.7
1.8
1.6
2.3
3.0
4.0
Operational Survey
Positive
Responses
3
11
1
12
Months
31
11
49
35
24
26
6
29
3
1
6
9
2
4
1
4
7
3
12
20
13
5
2
13
Percentage
Incidence
0.8
1.8
0.3
1.9
6.9
2.9
8.2
4.2
6.6
4.2
1.7
4.5
0.6
0.3
1.0
1.1
0.5
0.6
0.3
0.6
1.5
0.8
2.0
2.4
3.6
0.8
0.6
2.0
(continued)
145
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TABLE 72. (continued)
Baseline Survey
Operational Survey
Positive
Responses
Percentage
Incidence
Positive
Responses
Percentage
Incidence
0.0-3.5 km
North
East
South
West
3.5-S.Okm
North
East
South
West
7
13
5
3
7
13
Pain in Chest on Deep Breathing
0.7 8
1.7 4
0.9 13
1.6 11
1.6 7
0.6 5
1.8 4
1.7 6
Vomiting
1.8
1.1
2.2
1.3
1.9
0.8
1.1
0.9
0.0-3.5 km
North
East
South
West
3.5-5.0 km
North
East
South
West
8
21
16
17
7
7
9
30
2.0
4.4
2.1
2.1
2.3
1.4
2.3
4.0
13
2
15
13
14
15
3
15
2.9
0.5
2.5
1.5
3.8
2.4
0.8
2.3
For the responses to nausea, an increase was noted in the relative frequency along the south side and on
the west, a slight decrease in the north and a large decrease in the east. At the farther distances, the only
increase observed was along the northerly direction. The results for this response may also have been affected
by the combining of the two lowest distances used in the incidence analysis, since the 0 to 2.0 km data showed
an increase and the 2.0 to 3.5 showed a decrease. The total percentage incidence for nausea at these ranges
combined is nearly equal for the two periods as a result.
The responses for skin disease evidence a pattern which may indicate a treatment plant effect. The north
and south regions show an increase in percentage incidence at the close distance while the east and west
regions show a decrease. At the greater distances, the pattern is reversed, which indicates that the incidence
may not be related to the general welfare but may be specific to those narrow regions.
A similar pattern exists at the close range for diarrhea, with increases of 3.0 to 4.9 percentage incidence
for the north and south regions, respectively. At the greater distance, there was an increase in the north
region, while the south region showed a decreased incidence.
For pain in chest on deep breathing, an almost identical pattern was observed. The increase in percentage
incidence for north and south, respectively, were 1.1 and 1.3 for the 0 to 3.5 km data, while the east and west
frequency declined. The north region showed an increase at the 3.5 to 5.0 km range, but the percentage
incidence in the south declined as before.
The possible effect of age differences on the observed results were considered in view of the fact that the
age-distance relationship was shown to be significantly different between the two groups. The age groups
146
-------
determined by the physician to have the highest relative incidence were inspected. Of the nine diseases or
symptoms for which a distance effect was noted, children were reported to have the highest relative incidence
in five of the cases, no age group in three of the cases, and one (pain in chest on deep breathing) considered
not related.
By inspection of the data, if age group differences were to affect the results of the distance investigation,
then the higher percentage incidence would be expected in the survey period for which the greater frequency
of children at the 0 to 2.0 km range was observed. From the data shown previously in this Section, there was a
higher frequency of children at the closest distance during the baseline survey. As a result, the higher frequen-
cies obtained in the operational period were contrary to the change that would be expected based upon age
differences between the two groups.
To further investigate the possibility of an age or age-sex effect on the observed results, the percentage
incidence by age and sex groups at the closest distance was calculated for each disease and symptom determin-
ed to be possibly indicative of a treatment plant effect. These are shown in Tables 73 through 80, and were us-
ed to indicate if a particular group was responsible for the increase or if it was evident in all ages and both
sexes.
TABLE 73. PERCENTAGE INCIDENCE OF INFLUENZA BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey
Operational Survey
Percentage
Difference
Male
0-5
6-18
1945
46-64
65 +
11.1
17.1
6.7
3.6
0.0
14.6
14.5
11.8
5.7
0.0
3.5
-2.6
5.1
2.1
0.0
Female
0-5
6-18
19-45
46-64
65 +
7.7
7.1
11.2
12.2
0.0
16.3
8.3
11.2
5.7
0.0
9.3
1.2
0.0
-6.5
0.0
TABLE 74. PERCENTAGE INCIDENCE OF VOMITING BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey Operational Survey
Percentage
Difference
Male
0-5
6-18
19-45
46-64
65+
6.2
0.0
0.0
0.0
0.0
5.5
4.8
0.0
2.9
0.0
-0.7
4.8
0.0
2.9
0.0
Female
0-5
6-18
19-45
46-64
65+
1.5
1.4
1.1
2.0
0.0
6.1
7.1
2.0
0.0
0.0
4.6
5.7
0.9
-2.0
0.0
147
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TABLE 75. PERCENTAGE INCIDENCE OF DYSENTERY BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey
Operational Survey
Percentage
Difference
Male
0-5
6-18
19-45
46-64
65+
1.2
0.0
0.0
0.0
0.0
1.8
1.2
0.0
0.0
0.0
0.6
1.2
0.0
0.0
0.0
Female
0-5
6-18
19-45
46-64
65+
1.5
0.0
0.0
0.0
0.0
2.0
0.0
0.0
0.0
0.0
0.5
0.0
0.0
0.0
0.0
TABLE 76. PERCENTAGE INCIDENCE OF GENERAL WEAKNESS BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey
Operational Survey
Percentage
Difference
Male
0-5
6-18
19-45
46-64
65+
0.0
0.0
0.6
0.0
0.0
1.8
2.4
2.9
0.0
0.0
1.8
2.4
2.3
0.0
0.0
Female
0-5
6-18
19-45
46-64
65+
0.0
0.0
1.1
4.1
0.0
0.0
2.4
2.0
0.0
0.0
0.0
2.4
0.9
-4.1
0.0
148
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TABLE 77. PERCENTAGE INCIDENCE OF NAUSEA BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey
Operational Survey
Percentage
Difference
Male
0-5
6-18
19-45
46-64
65+
2.5
0.0
0.0
0.0
0.0
3.6
3.6
2.2
0.0
0.0
1.1
3.6
2.2
0.0
0.0
Female
0-5
6-18
19-45
46-64
65+
0.0
2.9
1.7
4.1
0.0
0.0
3.6
5.3
0.0
0.0
0.0
0.7
3.6
-4.1
0.0
TABLE 78. PERCENTAGE INCIDENCE OF SKIN DISEASE BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age
Group
Percentage Incidence
Baseline Survey Operational Survey
Percentage
Difference
Male
0-5
6-18
19-45
46-64
65+
0.0
0.0
0.6
0.0
0.0
1.8
1.2
1.5
2.9
0.0
1.8
1.2
0.9
2.9
0.0
Female
0-5
6-18
19-45
46-64
65+
1.5
0.0
1.1
0.0
12.5
2.0
2.4
2.0
0.0
0.0
0.5
2.4
0.9
0.0
-12.5
149
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Tat TABLE 79. PERCENTAGE INCIDENCE OF DIARRHEA BY AGE
AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age Percentage Incidence Percentage
Group Baseline Survey Operational Survey Difference
Male
0-5
6-18
19-45
46-64
65+
7.4
5.7
3.4
1.8
1.1
14.6
6.0
5.9
5.7
0.0
7.2
0.3
2.5
3.9
-1.1
Female
0-5 6.2 16.3 10.1
6-18 4.3 4.8 0.5
19-45 2.2 8.5 6.4
46-64 4.1 2.9 -1.2
65+ 0.0 0.0 0.0
TABLE 80. PERCENTAGE INCIDENCE OF PAIN IN CHEST ON DEEP BREATHING
BY AGE AND SEX GROUP AT 0 TO 2.0 km DISTANCE
Age Percentage Incidence Percentage
Group Baseline Survey Operational Survey Difference
Male
0-5 0.0 1.8 1.8
6-18 0.0 3.6 3.6
19-45 0.0 2.2 2.2
46-64 0.0 2.9 2.9
65+ 11.1 0.0 -11.1
Female
0-5 0.0 0.0 0.0
6-18 1.4 1.2 0.2
19-45 0.6 2.0 1.4
46-64 2.0 0.0 -2.0
65+ 0.0 0.0 0.0
As can be seen, for those diseases and symptoms for which an increase was observed at the 0 to 2.0 km
range, the tendency was for this to be irrespective of age or sex group. The increases, indicated by positive dif-
ferences, occur across both sexes and several age groupings. The implication is, then, that the observed dif-
ferences between the two periods are not related to the difference in the age or age-sex composition of the two
surveys.
Summary-
The summary of findings from the health survey is contained in Table 81. The following observations on
the results can be made when considering the entire set of illnesses and symptoms. It was considered that peo-
ple aware of the purpose of the study might bias their responses to the questionnaire toward the indication of
health hazard by, consciously or unconsciously, recalling a higher incidence than they might have otherwise.
Although this cannot be ruled out as a possibility, the available evidence from the results does not indicate
150
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TABLE 81. SUMMARY OF HEALTH SURVEY ANALYSES
Illness,
Disease, or
Symptom
Asthma-Hay
Fever
Influenza
Worms
Vomiting
Dysentery
General
Weakness
Nausea
Sore Throat
Skin Disease
Diarrhea
Pain in Chest
on Deep
Breathing
Test for
Independence
Significant at
0.01
Significant at
0.01
Significant at
0.01
Significant at
0.01
Significant at
0.05
Significant at
0.05
Significant at
0.05
Significant at
0.05
Not significant
Not significant
Not significant
Physician's
Evaluation
Not related
Related
Possibly
related
Related
Related
Related
Related
Related
Possibly
related
Related
Not related
Distance
Relationship
None
Slight increase
at 0-2, decrease
overall
None
Significant in-
crease 0-2,
slight decrease
elsewhere
Slight increase
0-2, decrease
elsewhere
Significant in-
crease 0-2,
significant de-
crease elsewhere
Significant in-
crease 0-2,
significant de-
crease elsewhere
None
Significant in-
crease 0-2, no
change elsewhere
Significant in-
crease 0-2, no
change elsewhere
Significant in-
crease 0.2,
slight decrease
elsewhere
Direction
Relationship
-
Increase at
close distance
in south
-
Increase at
close distance
north and south
None
Increase at
close distance
north and south
Increase at
close distance
in south
-
Increase at
close distance
north and south
Increase at
close distance
north and south
Increase at
close distance
north and south
that this occurred. The health variables studied were not all considered relative to the treatment plant, but
only one of the unrelated illnesses, asthma-hay fever, showed a significant value on testing for independence,
and one other was selected based on the distance relationship of the standardized normal deviates. If
misleading information was being obtained, there would likely be a higher incidence of significant non-related
diseases or symptoms.
Secondly, the judgment as to whether a treatment plant effect may have occurred is based on the
breakdown of the sample into the three distance groupings. The respondees were not informed of this,
however, nor of the group to which they belonged. Thus, people living in the same inner section could be
151
-------
classified as either 0 to 2.0 km or 2.0 to 3.5 km, and if the results differ between these groups, the possibility
of a prejudice existing is not as high.
With regard to the diseases and symptoms for which a treatment plant effect may be indicated, there is
some consistency when they are considered as a whole. Vomiting, nausea, and diarrhea can be expected to
occur together in many instances and should show the same pattern. Dysentery is also related to one or more
of the above, as can be influenza.
The directional relationship investigation showed that there was an increase in the south in the inner sec-
tors for all the diseases and symptoms which the prior analyses had labelled as a possible health effect of the
treatment plant, and in the north for all but two. These were the two prominent wind directions for the area
during the operational period, and the consistency of these results gives some credence to the possibility that
the changes in the general health were related to the sewage treatment plant.
For none of the responses did the change appear to be isolated to a single age or sex group which might
have occurred because of the differences in the age-sex composition of the two surveys.
HUMAN SUBJECTS CHARACTERISTICS
Human Subject Participation by Sampling Period
Number of Human Subjects-
A total of 231 human subjects were recuited in October 1974 for participation in the study. In addition,
with the development of the Lexington Green Apartment complex subsequent to the initiation of the study,
an additional 51 participants were recruited from this area alone in October 1975. All of these participants
were to continue in the study to its completion and no further participants were recruited. Due to movement
of people from the area, some of the 282 participants did not complete the study. The number of participants
from whom some clinical specimens were received in each of the five periods is shown in Table 82.
I
TABLE 82. PARTICIPANTS BY GROUP AND SAMPLING PERIOD
Sampling
Period
October 1974
February 1975
October 1975
February 1976
October 1976
Original
Human Subjects
231
223
-
205
192
Additional
Human Subjects
_
-
51
-
34
Total
231
228
51
205
226
A total of 192 (83 percent) of the original human subjects completed all four sampling periods, and 34
(67 percent) of the additional human subjects completed both of their sampling periods.
Age and Sex Distribution-
The distributions of both the original and the additional human subject participants are summarized in
Table 83 for the initial and final sampling periods. Four age classes are used, 0-6, 7-18, 19-45 and 45 and over,
and the subjects are summarized by sex. The age was fixed at the time of recruitment so each participant
would remain in the same age class throughout the study. Also shown are the percentages relative to the total
sample, and these may be compared to the census data in Section 4 for consistency.
152
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TABLE 83. AGE AND SEX DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Age Group
Initial
Male
No.
%
Female
No.
%
Final
Male
No.
%
Female
No.
, %
Original Hainan Subjects
0-6
7-18
19-45
46+
TOTAL
22
33
40
15
110
9.5
14.3
17.3
6.5
47.6
28
25
52
16
121
12.1
10.8
22.5
6.9
52.4
19
27
33
13
92
9.9
14.1
17.2
6.8
47.9
22
20
43
15
100
11.5
10.4
22.4
7.8
52.1
Additional Human Subjects
0-6
7-18
19-45
46+
TOTAL
1
1
13
3
18
2.0
2.0
25.5
5.9
35.3
7
4
15
7
33
13.7
7.8
29.4
13.7
64.7
1
0
10
2
13
2.9
0.0
29.4
5.9
38.2
4
0
12
5
21
11.8
0.0
35.3
14.7
61.8
There was little change in the relative frequency of the age-sex groups from the initial to the final sampl-
ing. The differences ranged from 0.1 percent to 0.9 percent, and indicate that both groups were comparable in
their overall makeup. There was more variability in the additional participants from initial to final sampling,
but this was largely due to the smaller sample size involved.
The balance between male and female participants in the original group was good, while the females
predominated in the additional group. For the original subjects, frequency of children was, by design, high
relative to the census data, with frequencies in both the 7-18 and 19-45 classes low relative to the census data.
For the additional group, the male and female 19-45 classes contained most of the participants, but the
Lexington Green area was not considered to be representative of the general area in composition.
Distribution of Participants Sampled in October 1976
The human subjects remaining throughout the study had been questioned during their recruitment at the
beginning of the study regarding personal, socioeconomic, medical, and exposure characteristics. The
responses to the questionnaire are summarized below for the 226 participants completing the study.
Personal Characteristics-
The age and sex distribution of the participants is summarized above for the October 1976 sampling
period. The racial distribution is summarized in Table 84. The participants were predominantly white,
TABLE 84. RACIAL DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Race
Number
Percentage*
Negro
Oriental
White
Other
Unspecified
3
4
173
1
45
1.6
2.2
95.6
0.6
-
*Percent of questionnaires where race indicated.
153
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comprising 96 percent of those participants for whom the race was identified. This is consistent both with the
general area, as indicated by the census data, and with the proportions in the household health survey as a
whole.
Hair color distribution is summarized in Table 85. The dominant hair colors were brown, with 53 per-
cent, and blonde, with 34 percent. Marital status of the participants is summarized in Table 86. A total of 123,
or 54.4 percent, were married, and 96, or 42.5 percent, were single, with the majority of the single individ-
duals resulting from participants under 18 years of age.
TABLE 85. HAIR COLOR DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Color Number Percentage*
Brown
Black
Red
Blonde
Gray
Unspecified
120
14
5
77
9
1
53.3
6.2
2.2
34.2
4.0
-
*Excluding unspecified.
TABLE 86. MARITAL STATUS DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Marital Status Number Percentage
Single 96 42.5
Married 123 54.4
Divorced 3 1.3
Widowed 4 1.8
There were three questions concerned with the employment of the participants. Each was asked to state
their present employment status, their occupation, and the nature of the business of their employer. These
results are summarized in Tables 87 through 89. A total of 73 individuals, or 32.3 percent, stated that they
were employed full time, with the majority of the remaining individuals classified as students or housewives.
Only one was unemployed and two were retired.
TABLE 87. OCCUPATIONAL STATUS DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Occupational Status Number Percentage
Employed Full Time
Employed Part Time
Unemployed
Housewife
Student
Play-Nursery School
Pre-School
Retired
73
14
1
42
62
9
23
2
32.3
6.2
0.4
18.6
27.4
4.0
10.2
0.9
154
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TABLE 88. OCCUPATION DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Occupation Number Percentage*
Professional
Administrative
Technical
Clerical
Sales
Foreman/Craftsman
Laborer
Student
Volunteer Worker
Waiter/Waitress
Teacher
Housewife
Retired
Other/inappropriate response
16
13
14
12
6
7
6
68
1
1
9
44
2
27
8.0
6.5
7.0
6.0
3.0
3.5
3.0
34.2
0.5
0.5
4.5
22.1
1.0
-
*Excluding Other/Inappropriate
TABLE 89. EMPLOYER DISTRIBUTION OF HUMAN
SUBJECT PARTICIPANTS
Description of Employer Number Percentage*
Medicine
Public Assistance
Law
Education
Business
Industry
Government
Construction
Research
Retired
Home
Other/inappropriate response
5 •
3
1
64
34 .
25
1
3
1
2
43
44
2.7 •
1.6
0.5
35^2
18.7
13.7
0.5
1.6
0.5
1.1
23.6
-
*Excluding Other/Inappropriate
Professional and administrative classifications accounted for 14.5 percent of the total sample and 33.3
percent of those in full or part time employment. There was no dominant occupation listed. Business and
industry dominated the employer distribution with 59 of the participants falling in those two categories. This
represents 67.8 percent of the participants listed as employed full or part time.
Socioeconomic Characteristics-
There were two questions relating to the socioeconomic status of the participants. These involved the
educational level and the occupation of the head of the household only. There were 90 total households
among the 226 participants, and the occupations and frequency of occurrence are shown in Table 90. Forty
percent of this group was classified as in either professional or administrative positions, and another 18.9 per-
cent in the technical. The educational levels of the heads of household in Table 91 show that a total of 48.9
percent had college and that 11 percent had attended graduate school. The responses in these two categories
indicate that the population being sampled was above average in both education and occupational classifica-
tions.
155
-------
TABLE 90. OCCUPATION OF HEADS OF HOUSEHOLDS
Occupation Number Percentage
Professional
Administrative
Technical
Clerical
Sales
Foreman/Craftsman
Laborer
Teacher
Librarian
Retired
18
18
17
5
8
8
9
5
1
1
20.0
20.0
18.9
5.6
8.9
8.9
10.0
5.6
1.1
1.1
TABLE 91. EDUCATIONAL LEVEL OF HEADS OF HOUSEHOLDS
Level Number Percentage
8th Grade
High School Incomplete
High School Complete
College Incomplete
College Complete
Graduate School
1
6
17
22
34
10
1.1
6.7
18.9
24.4
37.8
11.1
Medical History-
The participants were questioned regarding their medical history prior to the study. They were asked to
state whether they had ever been diagnosed for a number of chronic illnesses, corresponding to those for
which information was obtained in the household health survey. The summary of positive responses and
percentage incidence of these chronic illnesses prior to the study are shown in Table 92.
The highest percentage incidences were for asthma-hay fever and sinusitis-bronchitis, affecting 17.7 and
11.9 percent of the sample, respectively.
Exposure characteristics-
To evaluate the exposure of the individuals both to the Egan plant and to other possible sources of health
hazard, the participants were questioned regarding nearness to potential sources, length of residence both at
TABLE 92. FREQUENCY OF CHRONIC ILLNESSES AMONG
HUMAN PARTICIPANTS
Prior Diagnosis of
Chronic Illness
Tuberculosis
Malignancies
Asthma-Hay Fever
Diabetes
Heart Condition
Hypertension
Sinusitis-Bronchitis
Arthritis-Rheumatism
Rheumatic Heart Disease
Thyroid Disease
Liver Disease
Kidney Disease
Number of
Positive Responses
3
3
40
3
8
12
27
13
2
7
1
2
Percentage
1.3
1.3
17.7
1.3
3.5
5.3
11.9
5.8
0.9
3.1
0.4
0.9
156
-------
their current address and in the same city or town, time spent away from the home, and cigarette smoking
habits. In addition, the age and sex of the individual was tabulated with distance from the plant, since certain
of the illnesses or diseases potentially arising from sewage treatment plant operation are expected to have
higher relative incidence among certain age groups or in a specific sex.
The location of the participants with respect to distance and direction from the Egan plant is summarized
in Table 93. The location of each household was determined to the nearest one-tenth kilometer and recorded.
The residences then were located within respect to direction from the plant using 36 ten-degree sectors, where
the center of sector 36 lay directly north of the center of the treatment plant.
TABLE 93. PARTICIPANTS BY DISTANCE AND DIRECTION
FROM EGAN PLANT
Direction
North
Northeast
East
Southeast
South
Southwest
West
Northwest
Sector
(X 10°)
34 to .02
03 to 06
07 to 11
12 to 15
16 to 20
21 to 24
25 to 29
30 to 33
Distance (km)
0.0 to 0.9
2
0
0
0
0
0
0
32
1.0 to 1.9
3
0
0
4
20
0
8
0
2.0 to 2.9
14
0
5
4
15
14
17
44
3.0 +
0
0
0
2!
0 •
9
7
7
The individuals falling within the 0.0 to 0.9 range actually represent a range of only 0.4 to 0.6 km from
the plant, and represent the Lexington Green area. The next closest participants lie at 1.4 km from the plant.
The majority of the participants lie on the northwest and south sides of the center of the plant, and the north-
east and east sectors combined have only five participants. The predominant wind directions for the area are
from the north and south, with winds seldom coming from the east. Thus, the selected participants resided
largely in both the high exposure and the low exposure regions.
The exposure of the participants to other sources of potential health hazard was noted by recording the
distance from the residence to expressways, multi-lane highways, and industrial operations. The distance
from the participants' residences to the nearest expressway is summarized in Table 94.
TABLE 94. DISTANCE FROM NEAREST EXPRESSWAY FOR
HUMAN SUBJECT PARTICIPANTS
Distance
Less than 2 blocks
2 blocks up to 1 mile
At least 1 mile
Number
2 .
59
165
Percentage
0.9
26.1
73.0
157
-------
There were only two individuals who lived within two blocks of an expressway, with the majority, 73 per-
cent, living one mile or more away. The responses for closeness to multi-lane highways are shown in Table 95.
TABLE 95. DISTANCE FROM NEAREST MULTI-LANE HIGHWAY
I OR HUMAN SUBJECT PARTICIPANTS
Distance
Less than 2 blocks
2 blocks up to 1 mile
At least 1 mile
Number
22
92
112
Percentage
9.7
40.7
49.6
There were 22, or 9.7 percent, within two blocks and a higher percentage in the 2 blocks to one mile
range. Still about one half of the participants lived one mile or more from this potential source of health
hazard. All of the individuals responded that they lived one mile or more from the nearest industry.
The age and sex of the participants is classified for each of the distances from the Egan plant,
given in kilometers. The summary is shown in Table 96. There was a fairly uniform frequency of par-
ticipants at the middle three distance ranges, and there was an adequate representation of individuals
at each of the age groups for those three distances.
TABLE 96. AGE,-SEX.-DISTANCE DISTRIBUTION OF
HUMAN SUBJECT PARTICIPANTS
Distance
(km)
0-1
1-2
2-3
3-4
4-5
Age Group
0-6
Male
1
6
7
6
Female
5
11
3
7
7-18
Male
5
14
8
Female
2
9
9
19-45
Male
10
15
5
12
1
Female
12
20
11
11
1
46+
Male
2
4
7
2
Female
5
5
6
4
Total
35
68
62
59
2
The length of residency of the individuals at the time of recruitment is shown in Tables 97 and 98
for first the address and then the city. The majority of the participants, about 60 percent, had been at
their present address between one and three years, and only 11 percent less than one year. The
residency within the city showed a similar pattern except that the one to three category decreased to
about 52 percent.
TABLE 97. YEARS OF RESIDENCY AT PRESENT ADDRESS FOR
HUMAN SUBJECT PARTICIPANTS
Years
Less than 1
1 to 3
3 or more
Number
25
135
66
Percentage
11.1
59.7
29.2
158
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TABLE 98. YEARS OF RESIDENCE IN THE CITY FOR
HUMAN SUBJECT PARTICIPANTS
Years Number Percentage
Less than 1
1 to 3
3 or more
24
117
85
10.6
51.8
37.6
The number of hours spent away from the home was requested to indicate the adequacy of using
the residence as the primary measure of exposure. The number of hours per day that the individual
could be expected to be more than two miles from home was recorded and is summarized in Table 99.
The percentage of participants expected to be away from the home no more than one hour per day
was 50.9 percent, while 35 percent were listed as eight hours or more.
TABLE 99. HOURS PER DAY MORE THAN 2 MILES FROM
HOME FOR HUMAN SUBJECT PARTICIPANTS
Hours Number Percentage
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
90
25
4
8
2
2
10
6
19
17
21
8
8
1
2
3
39.8
11.1
1.8
3.5
0.9
0.9
4.4
2.7
8.4
7.5
9.3
3.5
3.5
0.4
0.9
1.3
Cigarette smoking habits are summarized in Table 100. There were 47 current smokers and 28
former smokers among the participants, while approximately two-thirds of the group were
nonsmokers. The definition of a nonsmoker, as previously used, was an individual who had not smok-
ed as many as 100 cigarettes in his lifetime.
TABLE 100. CIGARETTE SMOKING AMONG
HUMAN SUBJECT PARTICIPANTS
Status
Current Smoker
Former Smoker
Non-Smoker
Number
47
28
151
Percentage
20.8
12.4
66.8
159
-------
Comparison to Household Health Data-
The participants were selected from the baseline household health survey and the two groups can be com-
pared for consistency. The heads of household for the participants showed higher educational levels and more
professional and administrative occupations than did the baseline survey. Racially, both groups were almost
exclusively white.
For exposure characteristics, the participants showed lower exposure to other potential sources of
health hazard. The household health data indicated a greater frequency of occurrence of individuals close to
expressways and highways, and much more closeness to industrial operations. The participants were all in
excess of one mile from the nearest industry.
Distributions of 100 Human Subjects Analyzed for Additional Viral Serology
Summary of Selected Human Subjects and Samples-
Selection of blood samples for this additional study was based on several criteria. First, a sufficient quan-
tity of blood was required for the analyses for two separate samples, October 1974 or 1975, and October 1976,
to allow a comparison of before and after for antibody production. The two October samples were selected to
minimize any seasonal variability in the blood antibody levels. Three groups of subjects were set up. The first
group was composed of the additional participants from the Lexington Green area, since their proximity to
the plant gave them the greatest exposure. The second group was obtained from original participants living
near the plant (within 2.3 km) in the prevailing downwind directions (north, east, and south). The third group
was composed of original participants living farther from the plant in the infrequent downwind directions
(west and southwest).
Twenty-eight of the Lexington Green participants had sufficient samples of blood from the two sampling
periods to allow the evaluation of antibody production. The remainder of the required participants were
selected based on sample availability and on obtaining a similar age-sex distribution to the Lexington Green
group. This permitted a more valid comparison between the groups to be made than if the samples were
merely chosen on a random basis.
The three groups, Lexington Green, original-close, and original-far, were labeled Groups I, II, and III,
respectively. The identification numbers of the participants in these groups are shown in Table 101. The total
participants for each group was 28, 39, and 33, respectively.
TABLE 101. SELECTED PARTICIPANTS-
ADDITIONAL SEROLOGICAL TESTS
Group I
(Lexington Green)
603
614
615
616
617
619
621
628
630
631
632
633
634
640
641
Group II
(original— close)
2
3
4
11
12
13
14
15
16
29
33
157
158
159
162
Group III
(original -far)
49
50
74
75
76
. 83
84
97
101
102
141
143
145
146
284
(continued)
160
-------
TABLE 101. (continued)
Group I
(Lexington Green)
642
643
644
645
646
647
648
649
650
651
652
653
654
Group 11
(orignal-close)
165
166
172
177
178
181
182
196
199
206
207
213
214
226
227
228
229
231
233'
241
242
244
260
500
Group III
(original-far)
290
294
312
324
325
372
374
386
391
441
443
445
446
457
464
465
504
510
Human Subject Group Distributions-
The characteristics of the participants are summarized in Tables 102 and 103. All members of Group I
resided just northwest of the treatment plant, Group II was located primarily south of the plant, and Group
III was mainly toward the northwest. For both Groups II and III, however, there was a wide range of direct-
ions which were used.
TABLE 102. CHARACTERISTICS OF PARTICIPANT GROUPS
Characteristic
Group 1
Group II
Group III
Distance from John Egan Plant
Aeration Basins (km)
Predominant Direction from Plant (deg)
Range of Directions (deg)
Presumed Exposure Level
Number of Participants
Current Smoker Status:
Yes
No
Air Conditioning:
None
Window
Central
No Response
Education (head of household):
8th Grade
HS Incomplete
HS Complete
College Incomplete
College Complete
Graduate School
0.3-0.8
348
305-358
High
28
10
18
1
0
26
1
0
1
8
4
9
6
1.4-2.3
177
95-105
125-135
165-185
345-15
Moderate
39
6
33
8
8
23
0
0
1
3
13
19
3
2.8-3.5
316
125-145
215-245
265-275
305-325
Low
33
9
24
10
15
8
0
0
3
7
10
9
4
161
-------
TABLE 103. AGE AND SEX DISTRIBUTIONS OF PARTICIPANT GROUPS
Age (yr) Male. Female Total
Group I Participants
(0.3-0.8 km)
Total
Group 11 Participants
(1.4-2. 3 km)
Total
Group III Participants
(2.8-3.5 km)
Total
0-6
7-18
1945
46+
0-6
7-18
19-45
46+
0-6
7-18
19-45
46+
0
0
9
2
11
2
0
8
5
15
3 .
0
9
2
14
2
0
10
5
17
4
0
13
7
22
5
0
8
6
19
2
0
19
7
28
6
0
20
11
. 37
8
0
17
8
33
A higher frequency of smokers was observed in Group I and also a higher frequency of central air- condi-
tioning. The frequency of air-conditioning could reduce the effect of the presumed higher exposure levels by
removing particles from the air being drawn into the house. Groups II and III, with 21 and 30 percent, respec-
tively, of participants without air-conditioned homes would have greater exposure to any hazard- bearing par-
ticles in the air outside the home.
Socioeconomically, the first two groups could be considered equivalent, based on the educational level of
the head of the household. In both groups, over 50 percent had completed college with some members attend-
ing graduate school.
The age and sex distributions show that equivalent male-female breakdowns were observed in all groups
as the percentage of males ranged from 38 percent in Group II to 42 percent in Group HI. The breakdown by
age shows that all groups were predominantly adults, aged 19-45. There was a greater frequency of children in
Groups II and III than in Group I.
CLINICAL SPECIMEN MICROBIOLOGY
Statistical Methodology
The statistical analysis of the clinical specimen microbiology had two major objectives: to determine
from the self- and seasonally-paired data whether the occurrence of any of the pathogens analyzed increased
in the operational sampling periods, and, if so, to determine whether such increases were related to exposure
factors such as distance from the sewage treatment plant, or to other factors such as participant
characteristics.
In order to determine whether the occurrence of pathogens in the clinical specimens had increased after
the plant began operating, the analytical results of the samples taken from each participant were matched
according to season (February or October) and compared for each of the bacteria, parasites and viruses
analyzed in the four sample media (feces, throat swabs, sputum, and blood). The comparison of the paired
isolation results from bacterial, parasitic, or viral analyses of feces, throat swab, or sputum samples yielded
only three possible outcomes, which were coded as —, 0, and +:
162
-------
— the pathogen was isolated in the baseline period sample, but was not isolated in the paired
operational period sample;
0 the paired baseline and operational period samples both contained the pathogen or both were
negative for the pathogen; and
+ the pathogen was isolated in the operational period sample, but not in the baseline period
sample.
Only paired samples showing a change, either an increase or decrease, over the two sets of sampling periods
were statistically analyzed; incomplete pairs were deleted. The resulting data were evaluated through a series
of sign tests, using two-sided levels of significance, to test the null hypothesis of no difference between the fre-
quency of pathogen isolations reported for the baseline and operational sampling periods. For those
microbiological analyses in which the number of pairs showing a change was greater than 10, the normal
distribution approximation to the binomial expansion was employed in calculating the probability levels. All
of the microbiological isolation data were analyzed in this way.
The viral serology analysis results from the blood samples were reported in viral antibody liter. For most
of these analyses, six two-fold serial dilutions were performed so seven titer levels were available, beginning at
< 10 (no test reaction observed at the lowest dilution level) and increasing by orders of two until the final
value, ^320 (test reaction obtained at the highest dilution level). These titer values were receded for com-
parison purposes as values from 0 to 6, which each difference of one representing a two-fold change in
dilution.
The three poliovirus analyses were conducted using only two dilution levels (10 and 100); this resulted in
three titer values ( <10, 10 and > 100). For these data, a change from <10 to 10 was considered to be of ap-
proximately the same order of magnitude as one from 10 to 2:100. Thus the three tilers were receded as 0, 1,
2, and comparisons made of these values. In all cases, the blood samples were paired according to participant
and season, and the difference scores were obtained by subtracting the baseline sample titer from the operat-
ional sample titer.
Three statistical methods were identified to analyze each set of viral antibody data paired by participant
and season to determine whether there was an increase in the operational sampling periods: the paired sample
t-test, the Wilcoxon one-sample signed ranks test, and the sign test. The distributions of the antibody dif-
ference scores were not normal. Rather, they were leptokurtic, with the frequency of 0 values being much
greater than that of surrounding positive and negative integer values. In addition, the titer measurement being
the lowest dilution at which a test reaction is observed, does not constitute a continuous variable. Despite
these distributional characteristics, the paired sample t-test was considered as a potential statistical method.
Snedecor and Cochran^47\ (pp. 132-133) suggest that data restricted to a small number of values, thereby in-
volving a substantial number of zeros and ties, might be subjected to an ordinary t-test, with a correction for
continuity. However, even with this possible justification for using the more powerful t-test, we decided that
the violations of n'ormality and continuity were too severe to warrant its use. (It should be noted that
preliminary comparisons showed the t-test results to be quite similar to the Wilcoxon test results; the pro-
bability levels for the t-test were slightly lower, but not enough to affect the inferences made.)
Applying the sign test to this non-normal, discontinuous data would result in no invalid assumptions.
However, the sign test was rejected because consideration of the observed differences as merely positive or
negative would result in too great a loss of sensitivity. Instead, the Wilcoxon one-sample signed ranks test was
used, with the inclusion of corrections for continuity and for ties. Following Brownlee's suggestion*45^
(p. 260), all pairs yielding zero differences were excluded from the analyses. The resulting sample sizes were
large enough (all were greater than 30) to calculate approximate normal deviates for the rank sums. All in-
ferences were based on two-sided tests of significance.
163
-------
The relationship of pathogen increases to distance was investigated in two ways. The first approach,
which was used for the 23 sets of additional serological analyses conducted, was to compare the difference
scores for three groups of subjects divided according to the distance and direction they lived from the plant
(presumably a measure of exposure). The second method of investigation was stepwise multiple linear regres-
sion performed on the organisms which showed evidence of an increase from the baseline to the operational
sampling periods.
The results from the 23 additional serological analyses conducted for the 100 participants making up the
three exposure groups were reported in viral antibody liter. The statistical methods considered for these multi-
ple group comparisons were the one-way analysis of variance, the Kruskal-Wallis H test, and the chi-square
test. Again, the viral antibody difference scores were not distributed normally and did not represent a con-
tinuous variable. The parametric one-way analysis of variance was considered inappropriate for these
reasons. The non-parametric chi-square test is perhaps the most valid statistical method for this non-normal
discrete data because no method assumptions are violated. However, the chi-square test was rejected because
it would have caused a loss of sensitivity; in order to get expected frequencies of at least 1.0 in each cell, it
would have been necessary to collapse across all positive and all negative differences. This would have caused
a loss of all information regarding the magnitude of observed differences, thereby severely reducing the sen-
sitivity of the test.
The statistical method chosen to make the multiple group comparisons was the Kruskal-Wallis H test.
This test, a multiple group extension of the Wilcoxon signed ranks test, is a non-parametric version of the
one-way analysis of variance. The difference scores were ranked in order of magnitude from negative to
positive; the final test statistic was H', which included a correction factor for ties in the data.
The second approach in the investigation of the pathogen-distance relationship involved stepwise multi-
ple linear regression. A regression was performed for each organism exhibiting an overall increase in
occurrence or in liter level in the operational sampling periods. This criterion was relaxed, however, in two
cases (for the bacteria Streptococcus-alpha and Streplococcus-gamma); Ihese iwo polenlial palhogens, while
not illustraling an overall increase, did show a significanl increase for Ihe nearby Lexinglon Green par-
licipanls, along wilh a decrease or no change for Ihe more dislant participanls. Since ihis mighl be indicative
of a more general relalionship belween dislance from Ihe planl and palhogen occurence, Ihese polenlial
palhogens were also regressed. It was also decided that any of the 23 viruses showing liter differences between
groups would be subjected to regression procedures lo belter explicate the plant-virus relationship.
The dependent variables regressed were the paired isolation differences (with values of + 1 for + Codes,
0 for 0 codes, and —1 for — codes) or the paired antibody liter difference scores for each selecied organism.
The paired differences over all analyzed parlicipants in both seasons constiluled Ihe number of dependenl
variable observaiions. The participant questionnaire dala (see Seclion 5), the distance and direction of each
participant residence from the sewage treatment plant, and the season were assembled as the independent
observation items to comprise Ihe observalion dala base. Regressor (independent) variables (candidates to
enter the regression equation) were constructed from the independent observation items as various
hypothelical explanalions for Ihe observed operalional period increase in Ihe dependent variable.
The same 34 regressor variables (candidates for the equalion) were used in each regression. They are
lisled in Table 104. Some of Ihe variables came direclly from Ihe participant data; others were generated
through iransformations in the regression program. The regressor variables were conslrucled lo fall into three
general categories: possible plant exposure factors, olher environmenlal factors, and personal factors. The
exposure group included distance from Ihe planl, hours spent at home each day, type of air-conditioning,
direclion from Ihe planl, and exposure, which was compuled as hours al home divided by dislance. These
variables, if Ihey enlered inlo Ihe regression equalion wilh Ihe appropriale coefficienls, could implicale the
plant as a health hazard. As Table 104 indicates, if the sewage trealmenl planl were Ihe source of Ihe
pathogen, one would expect the coefficients to be negaiive for dislance and air-condilioning and positive for
hours home and exposure (if these variables had sufficienl explanatory potential to enler Ihe regression equa-
164
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TABLE 104. POTENTIAL VARIABLES FOR REGRESSION EQUATION
Possible Plant
Exposure Factors
Distance
Hours Home
Exposure
Air Conditioning
Direction^ (7 levels):
North
East
Southeast
South
Southwest
West
Northwest
Hypothetical
Relationship
to Pathogens*
-
+
+
-
+
+
-
- .
-
Other
Environmental
Factorsf
Season:
Fall
Winter
Traffic Proximity
Personal Factors!
Age* (9 levels):
0-3 yr
4-6 yr
7-12 yr
13-18 yr
19-25 yr
26-35 yr
36^t5 yr
46-55 yr
56 yr and Older
Smoking Status:
Current Smoker
Former Smoker
Nonsmoker
Illnesses:): (4 levels):
Asthma
Bronchitis
Heart Disease
Arthritis
Education:): (5 levels):
High School Incomplete
High School Graduate
College Incomplete
College Graduate
Graduate School
Sex
*If the regression coefficient were to have the indicated sign, it would be consistent with the hypothesis that the
pathogen emanates from the sewage treatment plant.
fFactors suggesting causes other than the sewage treatment plant for the operational period increase.
$The levels applicable to an individual were set equal to 1 for his observation; the other levels were set equal to 0.
tion). The expected direction relationships varied, and were determined from the prevailing wind directions
given in Figure 12 for the period of Egan plant operation.
The other two groups of variables, which had no expected relationship to the sewage treatment plant,
were included as indications of non-plant factors which might be responsible for the observed pattern of
pathogen increases in the operational period. The other environmental factors included season (February vs.
October) and traffic exposure (a weighted combination of the distances from the closest freeway and
highway). The personal characteristics included sex, age (with nine levels), education (with five levels), smok-
ing status, and prior chronic illnesses (with four levels).
The regression equations were generated in a stepwise fashion according to the usual optimizing
algorithm; the "best" equations were chosen through evaluation of ^ (psi), which is equal to the log.o
transformed probability level of the F-ratio for each equation. All of the dependent variables, and most of the
regressor variables had only two or three possible values for a participant/season observation (for example,
the —, 0, + coded paired comparison results from the pathogen isolations were evaluated as —1,0, +1; and
regressor variables were constructed for each level of the multi-level factors with values of I/in group and
165
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0/not in group). Thus, strong predictive equations were not expected from the regressions. Rather, the equa-
tions were intended merely to assist in interpreting the patterns of pathogen increases that had been observed
in the paired data. That is, to determine whether the patterns were associated with the various plant exposure,
environmental, and personal characteristics. Also, even though the proportion of variance accounted for by
the equations (R2) was not expected to be large, most equations had enough degress of freedom (about 400),
that even relatively small F-ratios were quite significant.
Bacterial Isolations
Pathogenic and potentially-pathogenic bacterial isolates were sought from the feces and sputum
specimens and from the throat swabs. The sputum specimens were only analyzed for Mycobacterium tub-
erculosis; no positive isolates were obtained. The feces specimens and throat swabs were both analyzed for
each of the'following pathogens and potential pathogens:
Proteus
Pseudomonas
Salmonella
Shigella
Staphylococcus aureus
Staphylococcus epidermidis
Streptococcus-alpha
Streptococcus-beta
Streptococcus-gamma
Proteus, Pseudomonas, and Salmonella were the only pathogen isolates in the feces samples; fecal Strepto-
cocci were frequently isolated but are considered non-pathogenic. Staphylococcus aureus and the three
Streptococcus groups were the listed pathogens that were frequently isolated from the throat swabs. One
isolate each of Proteus and Pseudomonas was also found in the throat swabs.
All of the individual data is tabulated by participant and sampling period in Appendix H. Table A-9 of
this appendix contains the bacterial isolation results for Proteus, Pseudomonas, Salmonella, and
Enterobacter in each of the feces specimens. The bacterial isolation results for Streptococcus-alpha,
Streptococcus-beta, Streptococcus-gamma, and Staphylococcus aureus are presented in Table A-10 of
Appendix H. A summary of the isolation data for the frequently isolated pathogens and potential pathogens
is given in Table 105 for each sampling period. The preponderance of the feces samples were negative for
Proteus, Pseudomonas, and Salmonella. The alpha and gamma types of Streptococci were commonly found
in the throat swabs. Isdolation of Staphylococcus aureus and Streptococcus-beta in the throat swabs occurred
less frequently.
In addition to the three pathogenic bacteria summarized in the table, isolates of three non-pathogenic
bacteria—E. coli, fecal Streptococci, and Enterobacter—were also reported for the feces samples. These
bacteria were detected with much greater overall frequency than the pathogens; E. coli was detected in 93<7o of
the samples, fecal Streptococci in approximately 90% to 95% of the samples, and Enterobacter in about 45%
of the samples. No consistent patterns of change in incidence were observed in these non-pathogenic bacteria
between the baseline and operational periods.
166
-------
TABLE 105. SUMMARY OF POSITIVE BACTERIAL ISOLATIONS
IN FECES SAMPLES AND THROAT SWABS
Samples Positive
Period
Analyzed Isolations
Isolation
Percentage
Feces Samples
1
T
3
4
5
1
2
3
4
5
1
2
3
4
5
212
205
45
182
190
214
211
46
206
209
208
200
44
210
205
Proteus
26
31
5
8
23
Pseudomonas
10
10
1
2
7
Salmonella
0
0
1
0
5
12
15
11
4 '
12
5
5
2
1
3
0
0
2
0
3
Throat Swabs
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2 .
3
4
5
223
223
47.
202
220
223
223
47
205
225
223
' 223
47'
202
225
223
223
47
.184
220
Staphylococcus aureus
20
36
2
24
66
Streptococcus-alpha
186
183
35
116
175
Streptococcus-beta
30
30
3
32
44
Streptococcus-gamma
63
135
17
95
116
9
16
4
12
30
83
82
74
57
78
13
13
6
16
20
28
61
36
52
53
167
-------
The bacterial isolation results from the feces specimens and throat swabs were paired according to par-
ticipant and season; the results of their comparisons were coded as—(a change from positive to negative), 0
(either both were positive or both were negative) or + (a change from negative to positive). The results have
been tabulated in Tables 106 and 107, by season and paired comparison result; the results for the Lexington
Green group have been tabulated separately, as samples were obtained from those participants only in the
October sampling periods. The two forms of 0 difference are indicated by the corresponding letters in par-
entheses, N (both samples were negative) and P (both samples were positive).
The Proteus, Pseudomonas, and Salmonella isolation results from the feces samples are summarized in
Table 106. Most of the participants' samples were negative for these three pathogens, as the large values in the
0(N) column indicate. Proteus and Pseudomonas show more decreases overall, while Salmonella shows a
greater number of increases. There also seems to be a slight tendency for the number of positive results to be
greater in the October sampling periods, especially in the operational year.
The results from the analyses of Staphylococcus aureus, and Streptococcus-alpha, -beta, and -gamma
performed on the throat swab specimens are presented in Table 107. Inspection of the table indicates that
Staphylococcus aureus and Streptococcus-beta were detected much less often than Streptococcus-alpha and
-gamma. However, Staphylococcus aureus and Streptococcus-beta show evidence of an overall increase in
isolations. For Staphylococcus aureus, this is due to the large number of positive differences (45) in the
October sampling; the February comparisons show a greater number of decreases in isolations of this
bacteria. The increased isolation of Streptococcus-beta does not show such a differential pattern with season;
-beta was detected more frequently in both the February and October operational periods.
The two other types of Streptococcus bacteria (-alpha and -gamma) illustrate a different pattern of iso-
lation results. They were both much more common, particularly the alpha type which was isolated in almost
TABLE 106. COMPARATIVE ANALYSIS OF BACTERIAL ISOLATIONS IN FECES SAMPLES
Paired Comparison Results
-
0
(N)
(P)
+
Total
Pairs
Proteus
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: Significant Decrease, P=0.019
23
15
1
39
134
131
27
292
2
5
0
7
5
13
2
20
164
164
30
358
Pseudomonas
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: No Change
Original Participants- February.
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: Approaching Significant Increase, P=0.13
9
8
1
18
172
147
29
348
0
0
0
0
2
7
0
9
183
162
30
375
Salmonella
0
0
1
1
183
158
27
368
0
0
0
0
0
4
2
6
183
162
30
375
168
-------
TABLE 107. COMPARATIVE ANALYSIS OF BACTERIAL ISOLATIONS
IN THROAT SWABS
Paired Comparison Results
0
(N)
(P)
+
Total
Pairs
Staphylococcus aureus
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: Very Significant Increase, P=0.004
22
6
1
29
167
131
25
323
1
2
0
3
7
45
5
57
197
184
31
412
Streptococcus-alpha
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: Very Significant Decrease, P<0. 00001
56
20
1
77
3
1
0
4
130
158
22
310
8
4
8
20.
197
183
31
411
Streptococcus-beta
Original Participants -February
Original Participants— October '
Lexington Green Participants-October
Total Comparisons
Result: Very Significant Increase, P=0.00001
1
0
1
2
184
159
29
372
0
0
1
1
12
13
0
25
197
172
31
400
Streptococcus-gamma
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: No Change
' 57
31
3
91
32
36
5
' 73
80
50
9
139
28
67
14
109
197
184
31
412
90% of the throat swabs. Streptococcus-alpha shows a much greater number of decreases than increases
overall; the number of positive and negative differences are about the same for Streptococcus-gamma. Both
bacteria, however, show more increases than decreases for the Lexington Green participants (those living
closest to the sewage treatment plant).
As in the Staphylococcus aureus results, the data for these two Streptococcus bacteria evidence different
patterns for the two seasons. Streptococous-alpha, normally detected in the throat swabs, was found much
less in the February operational period than in the October period.
Streptococcus-gamma illustrates this same pattern, and also shows more positive differences in the
October operational period.
Non-pathogenic bacteria in addition to the listed pathogens were also isolated in the throat swabs.
Micrococcus was detected in approximately 60% of the samples; Bacillus and Diptheroid were detected in less
than 3% of the samples.
169
-------
All of the pathogen isolation data sets were statistically analyzed with the sign test. In every case but one
(Salmonella isolations in feces samples), the sample sizes were large enough to permit use of the normal curve
approximation in calculating the probability of obtaining the observed number of increases or decreases. The
inferences made on the basis of these tests (using two-sided levels of significance) were summarized in Tables
106 and 107.
Of the three bacteria analyzed in the feces samples, only Salmonella showed any evidence of increased
isolation in operational period samples. This increase only approached significance (P = .13), so it cannot be
interpreted as very strong evidence of the plant as a source of Salmonella. Proteus and Pseudomonas both
showed decreased isolation in the operational periods; the decrease in Proteus isolations was significant (P =
.019).
The sign tests conducted on the throat swab pathogen isolations revealed very significant increases for
Staphylococcus aureus and Streptococcus-beta isolations in the operational periods (P = .004 and P =
.00001, respectively). Streptococcus-alpha isolations, however, decreased very significantly in the operational
periods (P < .00001).
The bacteria showing significantly increased isolation in the operational periods (Staphylococcus-aureus
and Streptococcus-beta) were used as dependent variables in multiple regression procedures. Streptococcus-
alpha and -gamma were also regressed, because they showed increased isolation in the participants living
closest to the plant (Lexington Green), along with no change or decreased isolation for the more distance par-
ticipants. A summary of the results of these regressions is presented in Table 108. The regressor factors have
been categorized into three general groups: exposure factors, other environmental factors, and personal fac-
tors. Each exposure factor has been characterized as to whether it implicates the sewage treatment plant; these
characterizations have been made on the basis of the relationship expected between each exposure factor and
the pathogen isolations if the plant is indeed serving as a source of aerosolized bacteria and viruses. (The
expected direction of the relationship of each exposure variable to the pathogens was presented earlier in
Table 104). Finally, the implications of the individual exposure variables were weighed against each other to
determine the implications of the entire equation.
The regression equation generated for Staphylococcus aureus accounted for 15.2% of the variance in the
dependent variable; the F-ratio for the equation was very significant (P « .0001). As Table 108 shows, five
regressor variables were included in the equation: southeast direction, season, two levels of age (4-6 years and
7-12 years) and asthma. Inspection of the student t-values for each factor indicates that the direction variable
demonstrates a much stronger relationship than any of the other factors; there was a strong tendency for the
operational period increases in Staphylococcus aureus to occur in participants living southeast of the plant.
The prevailing winds are not from the northwest, however, suggesting that this residential pattern is probably
unrelated to the sewage treatment plant. None of the other variables in the equation bear any relation to the
sewage treatment plant, leading to a general lack of evidence concerning the plant as a health hazard for this
bacterium.
The best equation (according to the \l/ statistic) generated for Streptococcus-beta had a multiple coeffic-
ient of determination (R2) equal to .067; even though this indicates that the equation only accounts for a small
percentage of variance (about 7%), the F-ratio was still significant (P = .0004). Four exposure factors came
into the equation—distance, hours at home, air conditioning and southwest direction—but three of the rela-
tionships exhibited were in the opposite direction of the expected relationships (see Table 104). That is,
pathogen isolation increased as distance from the plant increased, decreased as hours at home increased, and
increased in homes with more efficient air oonditioning systems. These reversals of the expected relationships
provided negative evidence for the hypothesized plant exposure function of Streptococcus-beta. The fourth
exposure factor (southwest direction) showed the predicted relationship—there were fewer increases in isola-
tions for persons living southwest of the plant. Although this was expected (the wind seldom blows from the
northeast), it is only indirect evidence; in addition, the low t-value associated with this factor indicates that
170
-------
TABLE 108. SUMMARY AND INTERPRETATION OF REGRESSION EQUATIONS FOR BACTERIAL ISOLATIONS
Regression Equation
Pathogen Regressed
Slaphylococcus jureus
Streptococcus-beta
Streptococcus-alpha
Streptococcus-gamma
R2
0.152
0.067
0.157
0.144
Regressor Variables in Equation
Possible Plant
Exposure Factors
Direction-increased isolation
in southeast direction
Distance-increased isolation
with distance
Hours Imme-decreased
isolation with hours at home
Air conditioning-increased
isolation in air-conditioned
homes
Direction-decreased isolation
in southwest diiection
l:.\p .sure-increased isolation
wiih exposure
Direction-decreased isolation
in west direction
Distance -increased isolation
with distance
Exposure -increased isolation
with exposure
Air-conditioning-increased
isolation in air-conditioned
homes
Direction-decreased isola-
lation in west direction
Direction -decreased isola-
tion in northwest direction
Other Environmental
Factors
Season-increased isolation in
October operational period
Season— decreased isolation
in February operational
period
Season -increased isolation in
October operational period
Persona)
Factors
4-6 yr olds
Age-increased isolation in
7-1 2 yr olds
Asthma-decreased
isolation in asthmatics
Age-increased isolation in
36-45 yr olds
Smoking-dccreased isolation
in ex-smokers
Illnesses - decreased isolation
other diseases
Education-decreased isolation
in households with high school
incomplete
Age-increased isolation in
26-35 yr olds
Smoking -increased isolation
in current smokers
in asthmatics
Education-increased isolation
in households with high school
incomplete
Education -decreased isolation
in households with graduate
school education
Age-decreased isolation in
7-1 2 yr olds
Age -decreased isolation in
26-35 yr olds
Age-decreased isolation in
36-45 yi olds
Age-decreased isolation in
those 55 yr and older
Smoker -increased isolation
in current smokers
Illnesses -increased isolation
in persons wiUr bronchitis
Education -increased isolation
in households with high school
incomplete
Student
t-Value
6.75
2.17
2.73
-1.95
3.RO
-1.70
2.54
-1.94
2.11
-1.64
-1.43
-1.42
3.65
-3.92
- 3.62
1.98
2.47
1.66
-2.20
1.61
2.21
2.02
-2.39
-2.68
5.19
-2.51
-1.53
-2.14
-2.42 •
1.91
2.02
2.03
Evidence of Plant as a
Potential Health Hazard
Exposure
l-'actors
-
_
-
•t-
+
+
_
+
-
+
+•
Overall
Equation
None
Very Slight
Some
Slight
171
-------
the relationship is fairly weak. The various other personal factors in the equation contribute no evidence con-
cerning the plant, so the entire set of relationships implicates the plant only minimally, if at all.
The regression equation for Streptococcus-alpha included eight regressor variables, had a coefficient of
multiple determination equal to .157, and had a significant F-ratio (P « .0001). Two plant exposure
variables were used in the equation; both illustrated the expected relationships to the increases in isolations.
Isolations increased with the computed exposure variable (hours at home/distance) and decreased west of the
plant. Inspection of the corresponding t-values indicates that these relationships are fairly strong. However,
season also exhibits a strong relationship to isolations. Because an overall decrease in isolations occurred in
the operational periods, the seasonal effect is interpreted to mean that more decreases occurred in the
February operational period than in the October period.
Two of the most important variables in the Streptococcus-alpha regression are exposure and season.
Because the Lexington Green participants were sampled only in the October periods and contained an opera-
tional period increase, the exposure and season variables may simply be pointing to a "Lexington Green par-
ticipant" factor that represents something other than exposure or season. To examine this possibility, a
second regression was run, with the Lexington Green participant data excluded. The resulting regression
equation did not include exposure. Thus, the original participant isolations give no evidence of plant ex-
posure; it is only in comparing the increased isolations of Lexington Green participants with the decreased
isolations of the original participants that an exposure factor appears. However, the west direction, season,
and most of the personal factors remained in the equation, suggesting that these represented more general
patterns over all participants. In general, the equations provided some evidence for Streptococcus-alpha being
related to the treatment plant, although the effect is only evident on the participants living within 0.7 km of
the plant.
The regression equation on Streptococcus-gamma accounted for 14.4% of the variance and was signifi-
cant beyond the .001 level (P « .0001). A number of plant exposure variables came into the equation.
Three of these variables—exposure, west and northwest—were in the predicted direction; isolations increased
with exposure, and decreased for areas west and northwest of the plant. The two other exposure factors—
distance and air conditioning—did not exhibit the predicted relationships; the frequency of isolations in the
operational period increased slightly with distance, and for homes with efficient air conditioning systems. In
addition, as in the Streptococcus-alpha regression, there was a very strong seasonal effect (isolations increas-
ed in the October operational period), which may have resulted from including the Lexington Green par-
ticipants. A second regression in which the Lexington Green participants were omitted did not bring in either
distance or the computed exposure variable as equation variables. Rather, it included hours at home; isola-
tions increased with number of hours spent at home during the day. The other regressor variables retained
essentially the same relationships as those of the first equation, with season still showing by far the strongest
effect. Overall, the Streptococcus-gamma regressions provided slight evidence for the predicted plant-
exposure relationship.
In summary, then, the regressions of the two throat swab bacteria showing an overall increase in opera-
tional year isolations (Staphylococcus aureus and Streptococcus-beta) demonstrated no, or very little,
evidence of a plant-exposure pattern. The regressions of the other two throat swab bacteria did exhibit some
plant exposure effects, with the evidence for plant exposure linked to the Lexington Green participants.
Alpha- and gamma-hemolytic Streptococcus species are part of man's normal flora in the intestinal tract,
upper respiratory tract and skin, and of his environment (e.g., vegetation, insects, and animal feces) and do
not normally produce disease. Therefore, their presence in the vicinity of the wastewater treatment plant or in
the throat swabs is of little practical health concern.
172
-------
Parasite Isolations
Seven hundred eighty-nine feces specimens were analyzed for a variety of helminths (parasitic worms)
and protozoa. The following parasites and commensals were sought:
Helminth-Nematodes
Ascaris lumbricoides
Enterobius vermicularis
Hookworm
Strongyloides stercoralis
Trichostrongylus sp.
Trichuris trichura
Helrhinth-Cestodes
Diphyllobothrium latum
Hymenolepis nana
Taenia sp.
Protozoa
Chilomastix mesnili
Dientamoeba fragilis
Entamoeba coli
Entamoeba hartmanni
Entamoeba histolytica
Endolimax nana
Giardia lamblia
lodamoeba butschlii
Trichomonas sp.
Protozoa were found in eleven samples; none contained helminths. The protozoans identified and their
frequencies are Chilomastix mesnili (1), Entamoeba coli (4), Entamoeba hartmanni (1), Endolimax nana (1),
Giardia lamblia (3), and Trichomonas (1).
The parasite results for each feces sample is given in Table A-l 1 in Appendix H. These parasite data are
summarized by sampling period in Table 109. Only 0 to 2% of the participant samples contained parasites.
TABLE 109. SUMMARY OF POSITIVE PARASITE ISOLATIONS
IN FECES SAMPLES
Samples Positive
Analyzed Isolations
Isolation
Percentage
All Parasites*
Period 1
Period 2
Period 3
Period 4
Period 5
195
190
45
198
169
2
1
0
4
4
1
1
0
2
2
*Organisms isolated: Chilomastix mesnili, Endolimax nana, Entamoeba coli,
Giardia lamblia and Trichomonas.
173
-------
The paired comparison results for the parasite isolations in the feces samples were obtained and coded in
the same manner as the bacterial isolation comparisons discussed earlier. These results are summarized in
Table 110 according to the season in which the samples were taken; again, the Lexington Green participants
have been listed separately, as they were not part of the original participant group.
TABLE 110. COMPARATIVE ANALYSIS OF PARASITE ISOLATIONS
IN FECES SAMPLES
Results of Paired Comparisons
0
(N)
(P)
+
Total
Pairs
Parasites
Original Participants-February
Original Participants-October
Lexington Green Participants-October
Total Comparisons
Result: Borderline Significant Increase, P = 0.06
0
0
0
0
155
121
29
.305
1
1
0
2
2
3
0
5
158
125
29
312
"Organisms isolated: Chilomastix mesnili, Endolimax nana, Entamoeba coli, Giardia lamblia andTrichomonas.
Out of a total of 312 samples pairs, only nine individual samples gave positive results in the isolation
analyses. Five of these represented a positive difference between baseline and operational samples. There were
no negative differences. These results were evaluated through a sign test; they reached a borderline
significance level, using two-sided probability levels (P= .06). However, caution should be exercised in inter-
preting the relationship of the parasite isolations to the treatment plant. There were improvements in sample
collection procedures in Period 5, when three of the five isolation increases (+) occurred; these improved pro-
cedures may be responsible for the increase. Also, all five sample pairs showing an increase were obtained
from participants living upstream from the plant. Since parasites are generally obtained by swimming or
washing clothes in water contaminated with feces, the upstream residence makes it unlikely the Egan plant
effluent was the source of the parasites. Thus, the observed borderline significant increase in parasite isola-
tions may be unrelated to the sewage treatment plant.
Viral Isolations
The feces samples and the throat swabs were both analyzed for a spectrum of virus groups and associated
microorganisms:
Adenovirus Papovavirus
Arbovirus Picodnavirus
Arenavirus Picornavirus
Coronavirus Poxvirus
Herpesvirus Reovirus
Oncornavirus Rhabdovirus
Orthymyxovirus Other viruses,
Paramyxovirus Chlamydia and Ricksettsia
No viruses were found in the throat swabs. All 20 of the positive virus isolates in the feces samples were
Picornaviruses. The Picornavirus group includes Polioviruses, Coxsackieviruses, and Echoviruses (besides
the nasal mucosal Rhinoviruses).
174
-------
The Picornavirus isolation results are tabulated for all participants in A-12 in Appendix H, and sum-
marized by sampling period in Table 111.
TABLE 111. SUMMARY OF POSITIVE VIRAL ISOLATIONS
IN FECES SAMPLES
Samples Positive
Analyzed Isolations
Isolation
Percentage
Picornaviruses
Period 1
Period 2
Period 3
Period 4
Period 5
207 0
199 4
44 0
194 9
183 7
0
2
0
5
4
The Picornavirus isolation data for the feces samples were compiled in the same manner as the bacteria
and parasite isolation data. The results of the paired comparisons are presented in Table 112. Most of the
samples were negative, although inspection of the table indicates that the number of positive samples
increased in the operational periods.
TABLE 112. COMPARATIVE ANALYSIS OF VIRAL ISOLATIONS
IN FECES SAMPLES
Results of Paired Comparisons
0
(N)
(P)
+
Total
Pairs
Picornavirus
Original Participants-February
Original Participants-October
Lexington Green Participants— October
Total Comparisons
2
0
0
2
176
145
28
349
2
0
0
2
5
7
0
12
185
152
28
365
Result: Significant Increase, P = 0.016
The Picornavirus isolation comparison results were evaluated through a sign test, using the normal curve
approximation to the binomial, as the sample size (N = 14) was larger than 10. This evaluation indicated that
there had been a significant increase in Picornavirus isolations in the operational sampling periods (P = .016,
two-sided).
After showing that there was a significant overall increase in occurrence of Picornavirus, it was of
interest to determine whether this increase conformed to a plant exposure pattern. This question was
investigated through a stepwise multiple linear regression procedure, as had been the throat swab bacterial
isolations. The same regressor variables were used: potential plant exposure factors, other environmental fac-
tors, and personal characteristics. The resulting regression equation explained very little of the variability in
the dependent measure (R2 = .0334). However, it did have a significant overall F-ratio(P = .031).
175
-------
The "best" equation chosen through evaluation of the \{/ statistic is presented in Table 113. It contained
five regressor variables. Two of these were in the plant exposure category (exposure and northwest direction),
one was classified with the environmental factors (season), and two were personal characteristics (26-35 years
old and asthma). The two potential exposure factors exhibited relationships opposite in direction to those
predicted by an exposure pattern (isolations decreased with exposure and were greater northwest of the plant).
This of course provided negative evidence for the plant as a Picornavirus health hazard. The other variables in
the equation, although useful in characterizing the individuals for whom the positive changes occurred, were
unrelated to a sewage treatment plant effect. Thus, the overall equation provides no indication of a plant
exposure-virus isolation relationship,
Viral Antibody Titer
Two sets of viral serology tests were conducted on the blood samples. Eight originally-planned serologic
tests were conducted on all the participant blood samples to measure the antibody levels to the following eight
viruses by the indicated procedure:
Coysackievirus B-l by hemagglutination inhibition
Echoviruses 3, 7, 11, and 12 by hemagglutination inhibition
Poliovirus types I, II, and III by serum neutralization
A second set of 23 additional serologic tests was conducted on the October blood samples of 100 par-
ticipants who were classified according to residential distance and direction from the plant into three groups
and matched by age and sex (see Tables 102 and 103). These 23 additional tests were:
Adenoviruses 1, 2, 3, 4 and 5 by serum neutralization
Coxsackieviruses A-7, A-9 and A-16 by serum neutralization
Coxsackieviruses B-3, B-5, and B-6 by hemagglutination inhibition
Echoviruses 4, 8, and 33 by serum neutralization
Echoviruses 6, 13, 19, 21, 25, and 29 by hemagglutination inhibition
Reoviruses 1, 2, and 3 by hemagglutination inhibition.
Each viral serology test determined the concentration range of the viral antibody in a blood sample by a
titration procedure. Each test was conducted at a series of dilutions of the blood serum. Most tests used six
serial two-fold dilution levels: 1 in 10, 1 in 20, 1 in 40, 1 in 80, 1 in 160, and 1 in 320. Whereas most of the viral
antibodies tested reflected natural exposure to the virus, many of the poliovirus antibody levels measured
presumably resulted from poliovirus vaccinations. Thus, the poliovirus tests were conducted at two ten-fold
dilution levels: 1 in 10 and Tin 100. The serologic test results were reported as a viral antibody liter. The liter
is the reciprocal of the greatesl serum dilulion level which slill produces Ihe specified test reaction. For exam-
ple, an end point dilution of 1 in 40 is reported as a liter of 40. A liter of < 10 indicates lhat no lest reaclion
occurred al Ihe leasl dilulion (1 in 10). An anlibody liler of > 320 for mosl of Ihe lesls or of > 100 for polio-
virus indicales lhat the test reaction occurred at Ihe grealesl dilution. Consecutive integer codes were assigned
to the geometric series of liter levels lo facililale the data analysis:
Mosl Viral Anlibodies Poliovirus Anlibodies
Code
0
1
2
3
4
5
6
176
-------
TABLE 113. SUMMARY AND INTERPRETATION OF REGRESSION EQUATIONS
FOR VIRAL ISOLATIONS
Evidence of Plant as a
Regression Equation Regressor Variable in Equation Potential Health Hazard
Possible Plant Other Environmental
Pathogen Regressed R^ Exposure Factors Factors
Picornavirus 0.033
Personal Student Exposure Overall
Factors t-Value Factors Equation
None
Exposure-decreased isolation
with exposure
Direction-increased isolation
in northwest direction
Season-increased isolation in
October operational period
Age—increased isolation in
26-35 yr olds
Illnesses-decreased
isolation in asthmatics
-1.96
2.41
1.49
1.43
-1.60
-------
Thus a code difference of 1 corresponds to a two-fold liter level change for most viruses tested and to a
ten- fold liter level change for the polioviruses. The comparison of lesl resulls paired by participant and
season was made using the difference score of Ihe codes for ihe paired resulls. A difference score of 0 in-
dicales both paired results were the same, while a difference score of + 2 for most viral antibody tests
represents a four- fold increase in liter. Thus, Ihe polenlial range of difference scores was from —6 lo + 6 for
mosl of Ihe viral antibody tesls and from —2 to + 2 for Ihe poliovirus lesls.
The individual parlicipanl tilers on the eight original tesls lhal were conducled on all blood samples in all
periods is given in Tables A-13 and A-14 in Appendix H. Table 114 summarizes these antibody liler dislribu-
lions for Ihe comparable results. The distribulions of Ihe Coxsackie B-l and Ihe four Echoviruses are similar,
illuslraiing fairly wide liter ranges. These distribulions are all highly skewed, being dominaled by negative ( <
10) values. A geometric mean was computed for the tiler dislribulion for each lesl and period, using Ihe
accepled convenlion of replacing each < value wilh half Ihe deleclion limit. Since each liter geometric mean
was less lhan 10, Ihese means were not reported, because values below ihe delection level cannol be con-
sidered numerically meaningful.
The anlibody liler for ihe Ihree polioviruses had differenl dislribulional characteristics. The antibody
levels lo Ihese viruses were very rarely below the detection level, and in facl, values of 100 or grealer were by
far Ihe mosl common. For Ihese Ihree viruses, geomelric mean anlibody levels have been reported for each
sampling period and for Ihe total samples. In compuling Ihese means, lilers < 10 were sel equal lo 5 and
lhose>100 were sel equal lo 100. The means for ihe Lexinglon Green participants are noticeably different
from Ihose of Ihe original participant.
The baseline and operalional means seem lo be quile comparable in mosl cases. One exceplion occurs in
Ihe polioviruses lype I anlibody lilers for ihe original participant' Oclober samples. These means differ by
more lhan 8 in liler, wilh Ihe Period 5 mean being greater.
The antibody liler levels for each of Ihe 23 addilional tests conducted on both of the October blood
samples of 100 selected participants are given in Tables A-15 through A-19 in Appendix H. The summary
antibody liter dislribulions for ihe Ihree exposure groups are presenled for each of Ihese addilional lests in
Table 115. The anlibody liler levels delecled by these tesls were generally lower lhan for the eight tests con-
ducted on all the participant' sera. However, anlibody liler levels of 40 were seen on four of Ihese lesls: Cox-
sackie B-5, Echo 4, Echo 6, and Echo 13.
Table 116 summarizes Ihe difference scores from ihe paired comparisons of Ihe liters from the eighl
original lests. As in the previous lable, Ihe dala have been tabulaled by sampling season. The widesl range of
difference scores was from —4 lo + 5 for Echovirus 3. Most difference scores were 0, however, indicating ihe
operalional period and baseline period liler levels were Ihe same. (Excepl for Ihe poliovirus lesls, Ihe
preponderance of sample pairs showing no change came from samples having antibody tilers 10 bolh limes.)
The resull reported wilh each virus labulation is from the statistical analysis of the tolal parlicipanl dala.
The resulls of Ihe paired comparisons of Ihe dala from the additional tests on the three parlicipanl
groups are presented in Table 117. The difference scores have been tabulaled for each group. The range of
difference scores for these lesls is lypically smaller lhan for the original tesls; Ihe largesi difference, which
occurs in Ihe Coxsackievirus B-5 comparisons, is + 3. Some of Ihe virus groups have particularly low ranges
of differences. The Reovirus group, for example, shows only a few + 1 differences: Reovirus 2 had no dif-
ferences al all belween ihe baseline and operalional samples. Allhough ihe purpose of Ihese lesls was lo look
for differences among exposure groups, nol for overall changes, inspection of the table does suggest that the
antibody to some viruses demonstraled overall increases in Ihe operalional samples (e.g., see Cox-
sackieviruses A-7, A-9, A-16, Echoviruses 21, 29). However, most of the viruses do not show differenl pal-
lerns of antibody change for the three groups.
178
-------
TABLE 114. COMPARATIVE VIRAL ANTIBODY TITER DISTRIBUTION
FROM ORIGINAL TESTS ON ALL HUMAN SUBJECTS
Original
Participants
February
Coxsackievirus B-l
Echovirus 3 '
Echovirus 7
Echoviru s 1 1
Echovirxis 12
T'i'
Poliovirus Type I
Geometric Mean
Poliovirus Type II
Geometric Mean
Poliuvirus Type III
Geometric Mean
Antibody
Titer*
<1()
10
20
40
80
<10
10
20
40
80
160
2.320
< 10
10
20
40
80
160
•MO
10
20
40
80
160
<10
10
20
40
80
160
100
Titer
<10
10
>100
Titer
<10
10
>100
Titer
Period 2
157
14
7
3
0
125
22
5
4
6
3
0
146
12
3
1
1
1
.151
8
2
1
2
1
171
14
10
2
0
1
6
36
139
57. 5
1
46
137
55. 5
3
55
124
47.6
Period 4
159
17
2
3
0
121
23
9
7
2
3
0
145
12
5
2
0
0
148
10
4
0
1
2
172
20
5
1
0
0
3
38
140
58. 9
1
45
138
56.2
4
54
124
47. 4
Original
Participants
Octobe r
Period 1
153
8
4
0
1
137
24
12
3
2
2
1
164
12
3
0
1
0
164
11
1
2
2
0
170
26
2
0
0
1
4
35
127
57. 4
2
38
126
57. 1
4
50
1 12
46.6
Period 5
150
10
6
0
0
131
31
4
2
2
2
1
159
17
2
2
0
0
161
14
2
2
0
1
174
23
1
1
0
0
2
28
136
65.6
1
41
124
55. 8
4
49
112
47.3
Lexington
Green
Participants
Period 3
26
1
1
0
0
25
0
1
0
0
0
0
20
5
1
0
0
1
25
1
1
0
0
0
26
4
0
0
0
0
2
10
19
39.3
3
8
20
41.4
1
7
23
54. 1
Period 5
25
1
2
0
0
24
1
0
1
0
0
0
22
3
1
0
1
0
24
2
0
1
0
0
27
1
1
1
0
0
2
1 1
18
36. 5
3
9
19
38.4
1
7
23
54. 1
670
51
22
6
1
563
101
37
20
12
10
2
656
61
I 5
673
46
10
6
5
4
740
88
19
5
0
2
19
158
579
57. 5
1 I
187
.564
54.8
17
222
519
^Reciprocal of serum dilution.
H-letriagglutination-Inhibition; serum dilution capable ot" inhibiting four hemagglutinating units of virus.
t'tSerum Neutralization Test; serum dilution capable of neutralizing 100 TCIUcQ (tissue culture infective
doses, 50% effective) virus.
179
-------
TABLE 115. COMPARATIVE VIRAL ANTIBODY TITER DISTRIBUTION
FROM ADDITIONAL TESTS ON 100 HUMAN SUBJECTS
Croup I
Group II
Group III
Lexington
Green
Antibody
Titer*
Adenovirus l'
<10
10
Adenovirus 2
<10
10
Adenovirus 3
<10
10
Adenovirus 4
<10
10
. , . , 1 1
Adenovirus 5
<10
10
Coxsackievirus A-7
<10
10
20
Coxsackievirus A-9 '
<10
10
Coxsackievirus A- 16 '
<10
10
Coxsackievirus B-3
<10
10
20
Coxsackievirus B-5
<10
10
20
40
Coxsackievirus B-6
<10
10
20
Participants
Period 3
26
2
25
3
26
2
27
1
28
0
24
4
0
26
2
27
1
24
3
1
24
4
0
0
25
3
0
Period 5
24
4
27
1
25
3
25
3
26
2
22
3
3
22
6
25
3
23
4
1
27
1
0
0
25
3
0
Original
Participants
(near)
Period 1
39
0
38
1
39
0
39
0
37
2
36
3
0
38
1
38
1
37
2
0
35
3
1
0
38
1
0
Period 5
38
1
37
2
36
3
38
1
36
3
34
5
0
36
3
34
5
34
4
1
32
6
1
0
37
1
1
Original
Participants
(far)
Period 1
32
1
31
2
32
1
33
0
30
3
29
4
0
33
0
32
1
28'
4
1
31
2
0
0
32
1
0
Period 5
31
2
32
1
30
3
30
3
30
3
26
6
1
29
4
31
2
29
3
1
30
2
0
1
30
3
0
Total
Sample
190
10
190
10
188
12
192
8
187
13
171
25
4
184
16
187
13
175
20
5
179
18
2
1
187
12
1
*Reciprocal of serum dilution.
t Hemagglutination-Inhibition; serum dilution capable of inhibiting four hemagglutinating units of virus.
tt Serum Neutralization Test; serum dilution capable of neutralizing 100 TdD^Q (tissue culture infective
doses, 50% effective) virus.
(continued)
180
-------
TABLE 115. (continued)
Echovirus 4tT
Echovirus 6
Echovirus 8
1"
Echovirus 13
t
Echovirus 1 9
Echovirus 21
Echovirus 25
Echovirus 29
Echovirus 33
t
Reovirus 1
Reovirus 2
Reovirus 3
Antibody
Titer*
<10
10
20
40
< 10
10
20
40
< 10
10
20
< 10
10
20
40
< 10
10
<10
10
< 10
10
< 10
10
20
< 10
10
20
< 10
10
< 10
10
< 10
10
Croup I
Lexington
Green
Participants
Period 3 Period 5
25
2
1
0
25
3
0
0
26
2
0
25
3
0
0
26
2
27
1
24
4
24
4
0
26
2
0
28
0
28
0
28
0
25
3
0
0
26
2
0
0
26
2
0
28
0
0
0
26
2
23
5
23
5
25
3
0
25
3
0
28
0
28
0
28
0
Group II
Original
Participants
(near)
Period I Period 5
33
3
2
1
33
6
0
0
34
5
0
37
2
0
0
35
4
38
1
35
4
38
1
0
35
1
3
39
0
39
0
38
1
31
4
3
1
31
: ' 7
0
1
37
0
2
34
3
2
0
33
6
33
6
36
3
31
6
2
33
3
3
39
0 .
39
0
36
3
Group in
Original
Participants
(far)
Period 1 Period 5
28
3
1
1
29
3
1
0
31
2
0
29
3
1
0
32
1
32
1
33
0
29
4
0
29
3
1
33
0
33
• 0
33
0
27
4
1
1
30
3
0
0
32
1
0
29
3
0
1
31
2
31
2
33
0
29
3
1
31
1
1
31
2
33
0
33
0
Total
Samples
169
19
174
24
1
1
186
12
2
182
14
3
1
183
17
184
16
184
16
176
21
3
179
13
198
2
200
0
196
4
*Reciprocal of serum dilution.
tHemagglutination-Inhibition; serum dilution capable of inhibiting four hemagglutinating units of virus.
ttSerum Neutralization Test; serum dilution capable of neutralizing 100 TCID,-,, (tissue culture infective
doses, 50% effective) virus.
181
-------
TABLE 116. SUMMARY OF VIRAL ANTIBODY TITER COMPARISONS
FROM ORIGINAL TESTS ON ALL HUMAN SUBJECTS
Coxsackievirus B-l
by Hemagglutination-Inhibition
Original participants-February
Original participants-October
Lexington Green participants(October)
Total comparisons
Result: No Change
-5
-4
Results of Paired Comparisons
-3 -2 -1 0 +1 +2 +3
0 0 14 160 7
0 4 7 142 9
0 0 0 27 0
21
329 16
0 0
4 0
1 0
5 0
+ 4
Total
+ 5 Pairs
0 181
0 166
0 28
0 375
Echovirus 3
by Hemagglutination-Inhibition
Original participants-February 0
Original participants-October 0
Lexington Green participants(October) 0
Total comparisons 0
Result: No Change
16 145
18 113
1 26
16
23
0
35 284 39
1 1 0
4 1 0
000
181
165
27
373
Echovirus 7
by Hemagglutination-Inhibition
Original participants-February 0
Original participants-October 0
Lexington Green participants(October) 0
Total comparisons 0
Result: No Change
4 169 6
10 143 7
3 24 0
17
336 13
0 1 0
200
000
2 10
180
164
27
371
Echovirus 11
by Hemagglutination-Inhibition
Original participants-February 0
Original participants-October 0
Lexington Green participants(October) 0
Total comparisons
Result: No Change
0
0 1 0 7 163 7
0 1 2 4 146 8
0 0 0 0 25 2
11
334 17
200
2 10
000
1
0
180
164
27
371
Echovirus 12
by Hemagglutination-Inhibition
Original participants-February
Original participants-October
Lexington Green participants(October)
Total comparisons 0
Result: Significant Decrease, P = . 045
(continued)
0 0 1 14 176 8 000 0 199
0 0 5 16 165 10 200 0 198
000226 20000 30
0 0 6 32 367 20 200 0 427
182
-------
TABLE 116. (continued)
-2
Results of Paired Comparisons
-1 0 +1
+2
Total
Pairs
Poliovirus Type I
by Serum Neutralization
Original participants-February
Original participants-October
Lexington Green participants (October)
Total comparisons
Result: Significant Increase, P= .014
173
155
28
356
6
10
1
17
181
165
31
377
Poliovirus Type II
by Serum Neutralization
Original participants-February
Original participants-October
Lexington Green participants (October)
Total comparisons
Result: No Change
181
160
30
371
184
166
31
381
Poliovirus Type III
by Serum Neutralization
Original participants-February
Original participants-October
Lexington Green participants (October)
Total comparisons
Result: No Change
180
163
31
374
181
166
31
378
183
-------
TABLE 117. SUMMARY OF VIRAL ANTIBODY TITER COMPARISONS
FROM ADDITIONAL TESTS ON 100 HUMAN SUBJECTS
Adenovirus 1 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Results of Paired Comparisons
-2 -1 0 +1 +2
0 1
0 0
0 0
0
I
24 3
38 I
32 1
94 5
Total
Pairs
28
39
33
100
Adenovirus 2 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III ( Orininal participants-far)
Total comparisons
Result: No Group Difference
0 2 26
0 0 38
0 1 32
0
96
0 0
I 0
0 0
1
0
28
39
33
100
Adenovirus 3 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
0 I 25 2 0
0 0 36 3 0
0 0 31 2 0
I
92
28
39
33
100
Adenovirus 4 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
0 I 24 3 0
0 0 38 1 0
0 0 30 3 0
I
92
28
39
33
100
Adenovirus 5 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
0 26 2
0 38 I
I 31 I
1
95
28
39
33
100
(continued)
184
-------
TABLE 117. (continued)
Coxsackievirus A-7 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Coxsackievirus A-9 Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Coxsackievirus A-16 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Coxsackievirus B-3 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Coxsackievirus B-5 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Results of Paired Comparisons
-3
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
-2
0
0
0
0
0
0
0
0
0
0
0
0
0 .
0
0
0
0
0
0
-I
I
0
1
2
0
0
0
0
0
0
0
0
1
0
3
4
4
3
1
0
23
36
28
87
23
37
29
89
26
35
32
93
25
36
28
89
23
31
30
+1
2
3
3
8
5
2
4
11
2
4
I
7
2
2
2
6
1
4
1
_f2_
2
0
I
3
0
0
0
0
0
0
0
0
0
I
0
1
0
I
0
+ 3
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
I
Total
Pairs
28
39
33
100
28
39
33
100
28
39
33
100
28
39
33
100
28
39
33
84
100
(continued)
185
-------
TABLE 117. (continued)
Results of Paired Comparisons
-2
-I
0
+1
+2
Coxsackievirus B-6 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Echovirus 4 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Echovirus 6 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Echovirus 7 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
0
0
0
0
0
0
0
0
0
0
I
I
0
0
0
2
0
0
2
2
I
2
5
1
1
2
4
0
3
1
26
38
. 31 .
95
25
35
29
89
27
35
29
91
28
34
32
0
0
2
2
1
2
I
4
0
2
I
3
0
2
0
0
1
0
1
0
1
I
2
0
0
0
0
0
0
0
Total
Pairs
28
39
33
100
28
39
33
100
28
39
33
100
94
28
39
33
100
Echovirus 13 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: Borderline Significant Difference, P = . 07
0 3 25 0 0
0 0 36 I 2
0 2 29 I I
0 5 90 2 3
28
39
33
100
(continued)
186
-------
TABLE 117. (continued)
Echovirus 19 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-hear)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
Echovirus 21 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group in (Original participants-far)
Total comparisons
Result: No Group Difference
Echovirus 25 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No. Group Difference
Echovirus 29 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Results: Significant Group Difference, P = . 02
Echovirus 33 by Serum Neutralization
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Results: Approaching Significant Difference, P = . 18
Results of Paired Comparisons
-2
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
-1
0
2
0
2
0
0
0
0
0
1
0
I
1
0
2
3
0
1
2
0
28
33
32
93
24
34
31
89
27
38
33
98
27
32
29
88
27
35
31
+1
0
4
1
5
4
5
2
11
I
0
0
I
0
5
I
6
1
3
0
+2
0
0
0
0
0
0
0
0
0
0
0
0
0
2
I
3
0
I
0
Total
Pairs
28
39
33
100
28
39
33
100
28
39
33
100
28
39
33
100
28
39
33
93
100
(continued)
187
-------
TABLE 117. (continued)
Reovirusl by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Res.ult: No Group Difference
Results of Paired Comparisons
-2 -I 0 +1 +2
28
39
31
98
28
39
33
100
Reovirus 2 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
28
39
33
100
28
39
33
100
Reovirus 3 by Hemagglutination-Inhibition
Group I (Lexington Green participants)
Group II (Original participants-near)
Group III (Original participants-far)
Total comparisons
Result: No Group Difference
28
37
33
98
28
39
33
100
188
-------
The statistical analysis of the paired comparisons from the original eight tests attempted to determine
whether the presence of the antibodies to these viruses in human blood samples had increased in the opera-
tional year. Parametric statistics were not appropriate for this data, as the antibody data were not normally or
continuously distributed. Thus, the Wilcoxon one-sample signed ranks test was used, with the inclusion of
corrections for continuity and for ties. The results of these tests are reported in Table 116 with the
corresponding paired comparison results. The antibody to two viruses demonstrated significant changes
between baseline and operational samples. One of these was the antibody to Echovirus 12, which significantly
decreased in the operational period (P = .045); the other was poliovirus type I, which evidenced a significant
increase after the sewage treatment plant began operation (P = .014).
The poliovirus I difference scores were subjected to multiple linear regression procedures to examine
whether the pattern is consistent with plant exposure. The resulting equation, which is summarized in Table
118, has an R2 equal to .095 and a significant F-ratio (P « .001). It provides no evidence of the plant as a
health hazard. Two of the three potential plant exposure factors which came into the equation do not possess
the expected relationships to the antibody liter changes. That is, liters in the operational samples increased for
persons living in homes with air condilioning and increased in areas wesl of Ihe plant. The second direclion
variable, soulheasl, had been considered neulral, so ils presence does nol serve lo implicale Ihe planl eilher.
The Iraffic variable showed a fairly slrong relalionship lo Ihe liter increases; seemingly, Ihose individuals
living closer lo major highways or freeways were exposed more lo Ihis lype of poliovirus. The Ihree personal
faclors in Ihe equalion indicale lhal ihe anlibody lilers decreased more for persons 4 lo 6 years and 26 lo 35
years and increased for Ihose suffering from aslhma.
The analysis of Ihe 23 addilional serology lesls required comparison of Ihe anlibody changes observed in
Ihe Ihree exposure groups. Il was reasoned lhal if the planl aerosols were contribuling lo the presence of viral
antibody in the blood, the effect would be proportional lo exposure. Our microbiological aerosol sampling
and modeling has indicaled lhal ihe negalive exponential of distance from the Egan plant is good measure of
long- lerm cumulalive exposure lo ils microbiological aerosol. Based only on this distance measure, Ihe Lex-
inglon Green participants (Group I) were expected lo evidence the grealesl increase in viral anlibody tiler. The
possibilily lhat other planl exposure faclors mighl enler in (such as wind direction, air condilioning systems,
hours spenl al home) was also considered. For example, Ihe facl that the Lexington Green area consisls of
aparlmenl houses, whereas Ihe original parlicipanls were predominanlly from residenlial homes, might affecl
Ihe dislance-exposure relationship. Thus, it was desirable to determine not only whelher Ihere was an overall
difference among groups, bul also to delermine whal ihese differences were ihrough ihe use of planned com-
parisons. The chosen comparisons were Group I vs. Group III, and Group II vs. Group III.
As before, Ihe liter difference scores for these Ihree parlicipanl groups were nol normally dislribuled.
Therefore, ihe paramelric one-way analysis of variance was inappropriale. The Kruskal-Wallis H test (a non-
parametric test) was employed in its place; Ihis lesl also includes a correclion for lies in Ihe dala. The lesl was
applied lo each of ihe 23 sels of data; Ihe resulting H1 statislic was evaluated on ihe chi-square dislribulion
wilh 2 degrees of freedom. As indicaled by Ihe resull slatements in Table 117, only Ihe changes in anlibody lo
Echovirus 29 were significantly different from the Ihree groups (P = .02). There was a borderline significanl
group difference in Echovirus 13 anlibody (P = .07), and a difference approaching significance for Ihe
changes in anlibody lo Echovirus 33 (P = .18).
The planned conlrasls previously menlioned were conducled on Ihe Echovirus 29 anlibody data.
Nemenyi's procedure for making a posteriori paired comparison48 was employed lo conlrasl Group I with
Group III, and Group II with Group III. Neither of Ihese comparisons were significant. It was felt thai Ihe
Nemenyi procedure's lack of a correclion for lies decreased ils sensilivily, so a Kruskal-Wallis H1 slatislic (for
K = 2 groups) was calculaled for each of Ihe desired comparisons. This procedure found Ihere lo be no dif-
ference belween Groups I and III, and a borderline significant difference between Groups II and III (P .052).
Although no formal comparisons were conducted for the Echovirus 13 or 33 data, inspection of Ihese dala in
Table 117 indicaies that the same basic patlern of group differences is present. In each case, it is the Group II
189
-------
Regression Equation
Virus Antibody Regressed R^
Poliovirus Type I 0. 095
TABLE 118. SUMMARY AND INTERPRETATION OF REGRESSION EQUATIONS
FOR VIRAL ANTIBODY TITERS
Regressor Variables in Equation
Possible Plant
Exposure Factors
Air conditioning-increased
titer in air conditioned homes
Direction-increased titer in
southeast direction
Direction-increased titer in
west direction
Other Environmental
Factors
Traffic-increased titer with
exposure to traffic
Echovirus 29
0.338
Direction-deer eased titer in
southeast direction
Direction-decreased titer in
northwest direction
Traffic-decreased titer with
exposure to traffic
Personal
Factors
Age -decreased titer in
4-6 yr. olds
Age -decreased liter in
26-35 yr. olds
Illnesses -increased titer
in asthmatics
Age -decreased titer in
26-35 yr. olds
Age-increased titer in
56 yrs. and older
Illnesses -decreased titer
in those with arthritis
Illnesses-increased titer
in those with bronchitis
Education-increased titer
in households with grad-
uate school education
Student
t-Value
3. 28
3.31
2.87
3.21
-1.83
-2.64
1.61
-3. 15
-5.23
-3.20
-1.56
1.70
-2. 52
1.72
2.79
Evidence of Plant As A
Potential Health Hazard
Exposure Overall
Factors Equation
Very slight
-------
participants (close original participants) who show the greatest increases in antibody level. This suggests that
there may be some effect related to plant exposure, in that the closer original participants show more antibody
increase than those living further from the plant. However, this exposure effect is not present for the Lex-
ington Green participants, as they evidenced no more antibody level increases than did the distant par-
ticipants. This pattern may be related to some different life styles or personal characteristics, as previously
mentioned.
In an effort to determine what factors were contributing to this pattern of liter increase, the Echovirus 29
data from the 100 participants making up the three groups were regressed. For this particular regression, two
of the age levels (7 to 12 and 13 to 18) were made unavailable as potential regressor variables, because none of.
the participants fell into these categories. In addition, season was not used, as all of these blood samples had
been taken in October periods.
The resulting equation, also summarized in Table 118, explained 33.8% of the variance and had an F-
ratio significant beyond the .001 level. However, the equation provided no direct evidence of the plant as a
source of Echovirus 29. One of the direction variables (northwest) did come into the equation with the
expected relationship; tilers decreased in the area norlhwesl of Ihe plant. This relationship, while quile slrong
(l = -5.23), provides only indirecl evidence againsl Ihe planl. The other direction variable, soulheasl, had no
expecled relalionship lo Ihe virus; Ihus, Ihe decrease in areas soulheasl of Ihe planl provided no damaging
evidence.
As in the poliovirus regression, Ihe traffic variable came into this equation with a fairly strong relation-
ship (t =-3.20) lo changes in anlibody liter. For Ihis virus, however, anlibody tiler decreased wilh Iraffic
exposure. Two age variables came inlo Ihe equalion—liter decreased in 26 to 35 year olds and increased in
those 56 or older. Current illnesses also illuslrated a relalionship lo Ihe anlibody liler, as Ihe tilers increased
for those suffering bronchitis and decreased for those with arthritis. Finally, education showed a fairly strong
relationship (t = 2.79); anlibody lo Ihe virus increased more for Ihose having a graduale school educalion.
In general, ihen, Ihe serology analyses provided Hide evidence of Ihe sewage Ireatment plant as an
exposure source of virus. For the one virus exhibiting an overall increase in anlibody liler (poliovirus I), there
was no evidence at all of the expected planl exposure relationship to Ihe observed liler increases. The
Echovirus 29 results give slighl evidence of an exposure-related patlern in the original parlicipanls, but no
effecl is observed in Ihe closer parlicipanls residing al Ihe Lexinglon Green Apartments. In addition, medical
microbiologisls generally agree lhat at least a faclor of four increase in anlibody liler is necessary before a
seroconversion is said to have laken place indicaling an immune response lo an infeclion. In Ihe case of
Echovirus 29, only ihree persons mel Ihe minimum requiremenl for seroconversion, iwo of which were from
Group II and Ihe olher from Group III. Considering all of Ihe dala, Ihe waslewaler irealmenl planl cannol be
implicaled with respect to Echovirus 29.
CLINICAL SPECIMEN TRACE METALS
The trace metal concentralions obtained on each individual participant's clinical specimens are presented
in the Individual Participant Data Appendix. These dala have been grouped by sampling period and by par-
licipanl age and sex for each irace melal (cadmium, copper, lead, mercury, and zinc) and for each specimen
lype (blood, feces, hair, and urine). Concenlralions for hair are based on dry weighl of Ihe sample, while for
feces wel weighl is used. Blood and urine are based on volume measuremenls. The summary slalislics which
have been compuled for each melal and specimen period-age-sex subgroup are Ihe geomelric mean, Ihe
arilhmelic mean, and Ihe siandard error (s/Vn). These summary slatistics were also compuled for Ihe sex,
age, and period marginal lolals. Summaries of Ihe cadmium concentration in blood, feces, hair, and urine
specimens are presented, respeclively, in Tables 119, 120, 121, and 122. The lead summary stalistics for each
specimen lype are given in Tables 123 - 126. Tables 127 -130 conlain Ihe mercury summaries, while Tables 131
- 134 presenl the summaries for copper and zinc. The blood hematocrits summary slalislics are also given in
Ihis seclion, in Table 135.
191
-------
TABLE 119. SUMMARY OF CADMIUM CONCENTRATIONS IN BLOOD
Cadmium Cone.
Sampling Period 1
Sampling Period 5
to
in mooa
Mg/100 ml
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
No. of
Subjects
171
11
52
79
29
87
7
31
35
14
84
4
21
44
15
Geometric
Mean
0. 078
0. 070
0. 070
0. 087
0. 074
0. 076
0. 080
0. 077
0. 082
0. 062
0. 080
0. 054
0.061
0. 092
0. 087
Arithmetic
Mean
0. 092
0. 073
0. 082
0. 101
0.091
0. 089
0. 083
0. 091
0.095
0. 071
0. 095
0.055
0. 068
0. 106
0. 110
Standard
Error
0. 004
0. 007
0. 007
0. 007
0. 014
0. 006
0. 008
0.011
0. 009
0. Oil
0. 007
0. 006
0.007
0. 009
0. 024
No. of
Subjects
169
34
40
70
25
80
13
23
31
13
89
21
17
39
12
Geometric
Mean
0. 095
0. 083
0. 089
0. 102
0. 106
0. 100
0.090
0.088
0. 108
0. 115
0. 091
0. 079
0. 089
0. 097
0. 098
Arithmetic
Mean
0. 108
0. 094
0. 102
0. 117
0. 116
0. 115
0. 101
0.097
0. 129
0. 128
0. 102
0. 091
0.108
0. 107
0. 102
Standard
Error
0. 004
0. 008
0. 009
0. 008
0. 009
0. 007
0.013
0. 009
0. 014
0. 015
0.006
0. Oil
0.019
0.008
0.009
-------
Cadmium Cone.
TABLE 120. SUMMARY OF CADMIUM CONCENTRATIONS IN FECES
Sampling Period 1 Sampling Period 5
Wet Weight
Age
Human Subjects
Total
0-6
7-18
19-45
46+
Males
Total
0-6
7-18
19-45
46+
Females
Total
0-6
7-18
19-45
46 +
No. of
Subjects
196
38
51
77
30
95
15
31
35
14
101
23
20
42
16
Geometric
Mean
0. 137
0. 154
0. 160
0. 124
0. 118
0. 135
0. 117
0.154
0. 133
0. 120
0. 140
0. 185
0. 170
0. 118
0. 116
Arithmetic
Mean
0. 158
0. 173
0. 177
0. 146
0. 134
0. 154
0. 128
0. 171
0. 156
0. 135
0. 162
0. 201
0. 185
0. 139
0. 133
Standard
Error
0. 0098
0.0131
0.0106
0.0112
0.0124
0.0088
0. 0147
0. 0136
0.0183
0. 0182
0.0085
0.0173
0.0174
0. 0131
0. 0174
No. of
Subjects
171
31
43
71
26
84
15
24
32
13
87
16
19
39
13
Geometric
Mean
0. 0906
0. 1202
0. 0873
0. 0873
0. 0763
0.0930
0. 1343
0. 1002
0. 0806
0. 0753
0. 0884
0. 1083
0. 0733
0.0931
0. 0773
Arithmetic
Mean
0. 1250
0. 1585
0. 1257
0. 1176
0. 1040
0.1288
0. 1747
0. 1409
0. 1100
0. 0995
0. 1213
0. 1433
0. 1065
0. 1238
0. 1085
Standard
Error
0.0071
0.0196
0. 0148
0.0104
0. 0138
0.0110
0. 0270
0.0232
0. 0167
0.0194
0. 0090
0.0286
0.0159
0. 0130
0.0204
-------
TABLE 121. SUMMARY OF CADMIUM CONCENTRATIONS IN HAIR
Cadmium Cone.
in Hairing/g
Dry Weight
Human Subjects
Sampling Period 1
Sampling Period 5
Males
Females
Age
i
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
No. of
Subjects
229
49
59
89
32
108
22
34
37
15
121
27
25
52
17
Geometric
Mean
0.607
0. 933
0. 957
0.364
0.564
0.689
0.821
0.872
0. 509
0.656
0.542
1. 033
1.088
0.286
0.494
Arithmetic
Mean
1. 086
1.376
1.386
0.745
1. 035
1. 167
1.383
1.365
0.871
1. 134
1.012
1.370
1.415
0.655
0.947
Standard
Error
0. 086
0.217
0. 188
0.098
0.234
0. 144
0.430
0. 294
0. 143
0.337
0.099
0. 191
0. 203
0. 134
0.332
No. of
Subjects
190
41
46
75
28
92
19
27
33
13
98
22
19
42
15
Geometric
Mean
0. 431
0.824
0.550
0.273
0.381
0. 57 1
0. 725
0.635
0. 486
0.489
0.332
0.921
0.449
0. 174
0. 306
Arithmetic
Mean
0.823
1.037
0.803
0.619
1.086
0. 910
0.958
0.965
0.886
0.784
0.741
1. 105
0. 574
0.409
1.349
Standard
Error
0. 097
0. 108
0. 132
0. 123
0.506
0.101
0. 170
0.206
0. 192
0. 198
0. 163
0. 139
0. 112
0. 153
0. 940
-------
TABLE 122. SUMMARY OF CADMIUM CONCENTRATIONS IN URINE
vo
on
Cadmium Cone.
in Urine
wg/t
Human Subjects
Mates
Females
Age
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
No. of
Subjects
225
45
58
90
32
106
20
34
37
15
119
25
24
53
17
Sampling
Geometric
Mean
0. 591
0. 458
0. 526
0. 645
0. 812
0.611
0. 444
0. 575
0.615
1. 056
0. 573
0. 470
0. 464
0.667
0.644
Period 1
Arithmetic
Mean
0. 742
0.630
0. 630
0.789
0.971
0.781
0. 723
0.696
0. 715
1. 213
0.708
0. 555
0. 536
0. 841
0. 758
Standard
Error
0. 039
0. 107
0. 052
0. 062
0. 100
0. 062
0. 231
0. 079
0.072
0. 165
0. 048
0. 059
0. 055
0. 092
0. 096
No. of
Subjects
190
40
47
75
28
91
18
27
33
13
99
22
20
42
15
Sampling
Geometric
Mean
0. 488
0.405
0.316
0. 589
0. 794
0.502
0.396
0. 340
0. 664
0. 773
0. 475
0. 413
0. 286
0. 537
0. 813
Period 5
Arithmetic
Mean
0.638
0. 579
0. 416
0.716
0.885
0.671
0.615
0.449
0.807
0. 868
0.606
0. 549
0.371
0.644
0. 900
Standard
Error
0. 031
0. 070
0. 045
0. 048
0. 072
0.048
0. 115
0.065.
0.086
0. 100
0.039
0.087
0. 060
0.053
0. 107
'
-------
TABLE 123. SUMMARY OF LEAD CONCENTRATIONS IN BLOOD
Lead Cone.
in Blood
ug/100 ml No' °f
Age Subjects
Human Subjects
Total 206
0-6 27
7-18 57
19-45 90
46+ 32
Males
Total 101
0-6 13
7-18 34
19-45 39
46+ 15
Females
Total 105
0-6 14
7-18 23
19-45 51
46+ 17
Sampling
Geometric
Mean
10. 30
10.94
9.44
10.20
11.75
11.61
11.07
9.73
13.25
12.82
9.18
10.82
9.04
8.35
10.88
Period 1
Arithmetic
Mean
11. 11
11.47
9.99
11.22
12.51
12.37
11.43
10.35
13.99
13.55
9.90
11.51
9.46
9.09
11.59
Standard
Error
0.31
0.68
0.46
0.51
0.83
0.44
0.77
0.65
0.75
1.24
0.39
1.13
0.62
0. 54
1. 11
No. of
Subjects
199
26
*6
88
29
94
11
33
37
13
105
15
23
51
16 ,
Sampling
Geometric
Mean
10.67
11. 12
9.76
10.78
11.85
11.88
11.60
9.80
14.07
12.27
9.69
10.78
9.71
8.88
11.53
Period 2
Arithmetic
Mean
11.42
11.61
10. 13
11.86
12.41
U.70
12.25
10.13
15.03
12.95
10.28
11.15
10. 13
9.57
11.97
Standard
Error
0.32
0.68
0.36
0.60
0.74
0.50
1.30
0.46
0.92
1.22
0.38
0.73
0.60
0.61
0.93
No. of
Subjects
171
35
40
70
26
80
13
23
31
13
91
22
17
39
13
Sampling
Geometric
Mean
9.78
9. 53
9. 11
9.98
10.72
10.81
8.87
9.84
12.30
11.45
8.96
9.94
8.22
8.45
10.05
Period 5
Arithmetic
Mean
10.30
9.84
9.39
10.68
11.29
11.35
9.32
10.08
12. 82
12. 12
9.38
10. 15
8.46
8.99
10.47
Standard
Error
0. 26
0.43
0.37
0.46
0.76
0.40
0.84
0.49
0.65
1. 17
0.30
0.47
0.49
0.51
0.98
-------
TABLE 124. SUMMARY OF LEAD CONCENTRATIONS IN FECES
Lead Cone.
in Feces,tig/g
Wet Weight
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
No. of
Subjects
208
41
54 .
82
31
98
.16
32
36
14
110
• 25
22
46
17
Sampling
Geometric
Mean
0.956
1. 169
1. 102
1. 084
0.699
0.958
0.900
1.236
0. 927
0.626
0. 954
1.384
0.933
0.854
0. 765
Period 1
Arithmetic
Mean
1. 206
1.591
1. 275
0. 885
0. 901
1. 155
1.036
1. 471
1. 093
0. 729
1. 252
1.946
0.987
1. 077
1. 043
Standard
Error
0. 072
0. 283
0. 097
0.075
0. 133
0.070
0. 135
0. 146
0.099
0. 102
0. 121
0. 446
0. 075
0. Ill
0. 225
No. of
Subjects
166
28
41
71
26
81
13
23
32
13
85
15
18
39
13
Sampling
Geometric
Mean
0.581
0. 755
0.658
0. 513
0. 505
0. 553
0.856
0.654
0.429
0.496
0.609
0.678
0.664
0. 594
0. 513
Period 5
Arithmetic
Mean
0. 723
0.891
0.784
0.669
0. 592
0.695
0.985
0.797
0.546
0. 595
0.749
0. 809
0.769
0. 771
0.588
Standard
Error
0. 039
0. 089
0.074
0.066
0. 068
0. 051
0. 121
0. 113
0. 065
0. 109
0.058
0. 129
0. 090
0. 105
0. 188
-------
TABLE 125. SUMMARY OF LEAD CONCENTRATIONS IN HAIR
00
Lead Cone.
in Hair,(ig/g
Dry Weight
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
No. of
Subjects
228
49
58
89
32
107
22
33
37
15
121
27
25
52
17
Sampling
Geometric
Mean
9. 44
16. 11
11.01
6.32
9.60
11. 15
15. 15
10. 93
10.63
8.34
8. 15
16.93
11. 12
4.37
10. 86
Period 1
Arithmetic
Mean
17. 83
22. 10
16. 14
14.69
23.09
17.88
24. 77
18. 28
15.48
12. 78
17.79
19.92
13.31
14. 12
32. 19
Standard
Error
2. 54
4.60
3. 57
3.73
11. 44
2.90
9.77
6.17
2.49
2.61
4.05
2.77
1.64
6. 16
21.48
No. of
Subjects
190
41
46
75
28
92
19
27
33
13
98
22
19
42
15
Sampling
Geometric
Mean
7.36
12.32
7. 75
5. 06
8.66
9.75
10.77
9.90
8.66
1 1 . 03
5.65
13.84
5.47
3.32
7. 02
Period 5
Arithmetic
Mean
14.36
15.25
11.62
10.25
26. 17
17.98
13. 78
14. 91
IS. 14
37.72
10.28
16. 52
6.94
6.41
16. 17
Standard
Error
2. 12
1.39
2. 17
1.76
12. 78
4. 07
1.63
3.48
3. 23
26.84
1.36
2. 18
1.25
1.68
6. 11
-------
TABLE 126. SUMMARY OF LEAD CONCENTRATIONS IN URINE
Lead Cone.
in Urine
ug/l
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
No. of
Subjects
225
45
58
90
32
106
20
34
37
15
119
25
24
53
17
Sampling
Geometric
Mean
8. 49
9. 27
8. 07
8.72
7.62
9. 00
9.32
8.31
9.41
9. 19
8. 06
9.23
7. 75
8. 26
6.45
Period 1
Arithmetic
Mean
9. 97
10. 53
9. 14
10. 63
8.83
10. 78
11. 12
9. 78
11. 81
10. 08
9. 24
10. 06
8. 24
9. 81
7. 72
Standard
Error
0. 38
0. 78
0.62
0. 71
0. 77
0. 87
1.48
0. 95
1. 29
1. 19
0. 44
0. 78
0.64
0. 80
0. 96
No. of
Subjects
190
40
47
75
28
91
18
27
33
13
99
22
20
42
15
Sampling
Geometric
Mean
4. 84
5. 10
4. 03
5. 15
5. 17
5. 07
4. 37
3.74
6. 48
6. 26
4. 64
5. 80
4. 47
4. 30
4. 38
Period 5
Arithmetic
Mean
6. 18
6.27
5.33
6.54
6. 48
6. 43
5. 14
4. 90
7. 74
8. 02
5. 95
7. 19
5. 91
5. 60
5. 15
Standard
Error
0. 30
0. 68
0. 51
0. 47
0. 97
0. 45
0. 60
0.62
0. 73
1. 83
0. 41
1. 12
0. 86
0. 58
0. 80
-------
TABLE 127. SUMMARY OF MERCURY CONCENTRATIONS IN BLOOD
.Mercury Cone.
in Blood
pg/100 ml
Age
Human Subjects
Total
0-6
7-18
19-45
46 +
Males
Total
0-6
7-18
19-45
46 +
Females
Total
0-6
7-18
19-45
46 +
No. of
Subjects
207
28
57
90
32
102
14
34
39
15
105
14
23
51
17
Sampling
Geometric
Mean
0.488
0. 725
0.359
0. 531
0. 471
0. 453
0.642
0.303
0. 537
0. 524
0. 525
0. 819
0.461
0. 527
0. 429
Period 1
Arithmetic
Mean
0. 752
1. 499
0. 578
0.696
0. 564
0.711
1.328
0. 513
0.686
0.650
0. 791
1.670
0.674
0. 704
0. 489
Standard
Error
0. 081
0. 539
0..070
0.049
0.057
0. 114
0.773
0.084
0.077
0. 102
0. 114
0. 777
0.120
0. 064
0. 055
No. of
Subjects
169
34
40
70
25
80
13
23
31
13
89
21
17
39
12
Sampling
Geometric
Mean
0. 198
0. 109
0. 183
0. 272
0. 208
0. 187
0. 135
0. 176
0. 232
0. 174
0. 208
0. 096
0. 194
0.308
0. 252
Period 5
Arithmetic
Mean
0. 267
0. 156
0. 228
0.329
0.309
0. 243
0. 172
0. 205
0. 297
0. 250
0. 289
0. 146
0. 259
0.354
0.374
Standard
Error
0. 016
0. 019
0. 025
0.023
0.065
0.017
0.030
0.023
0. 032
0. 043
0.026
0. 025
0.048
0.032
0. 127
-------
TABLE 128. SUMMARY OF MERCURY CONCENTRATIONS IN FECES
Mercury Cone.
in Feces,g/g
Wet Weight
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46 +
No. of
Subjects
207
41
55
80
31
98
16
32
36
14
109
25
23
44
17
Sampling
Geometric
Mean
0. 077
0. 041
0. 062
0. Ill
0. 097
0. 068
0. 033
0. 064
0. 102
0. 061
0. 086
0. 048
0. 059
0. 119
0. 143
Period 1
Arithmetic
Mean
0. 197
0. 100
0. 119
0. 167
0. 541
0. 126
0. 046
0. 124
0. 168
0. 114
0.261
0. 135
0. 112
0. 167
0. 893
Standard
Error
0.050
0.037
0.022
0. 017
0.325
0. 017
0. Oil
0.033
0. 031
0. 039
0. 094
0.060
0.027
0. 019
0. 586
No. of
Subjects
164
29
40
69
26
80
14
22
31
13
84
15
18
38
13
Sampling
Geometric
Mean
0. 066
0. 031
0. 056
0. 100
0. 663
0.058
0. 031
0. 063
0. 080
0. 044
0. 075
0. 031
0.049
0. 121
0. 091
Period 5
Arithmetic
Mean
0. 116
0.051
0.097
0. 158
0. 110
0. 104
0.052
0. 119
0. 130
0. 072
0. 128
0..051
0. 071
0. 180
0. 147
Standard
Error
0. Oil
0. 014
0. 020
0.018
0.022
0.015
0.021
0.034
0.026
0. 024
0.015
0. 020
0.014
0.025
0.035
-------
TABLE 129. SUMMARY OF MERCURY CONCENTRATIONS IN HAIR
o
Mercury Cone.
in Hair^g/g
Dry Weight
Human Subjects
Males
Females
A££
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46+
No. of
Subjects
228
48
59
89
32
107
21
34
37
15
121
27
25
52
17
Sampling
Geometric
Mean
0.638
0. 809
0.631
0.562
0.652
0. 547
0.710
0. 463
0. 511
0.654
0. 732
0.895
0. 960
0. 602
0.650
Period 1
Arithmetic
Mean
0. 994
1.046
1.492
0. 728
0.739
0.687
0.955
0.550
0.647
0.720
1.266
1. 116 .
2.774
0. 785
0. 757
Standard
Error
0.200
0. 102
0.762
0.059
0. 068
0.048
0.143
0.068
0. 080
0. 083
0.373
0. 144
1.784
0.084
0. 107
No. of
. Subjects
101
21
28
43
9
48
10
17
19
2
53
11
11
24
7
Sampling
Geometric
Mean
0.352
0.432
0.328
0.332
0.355
0.309
0.347
0.266
0.346
0.222
0.395
0.527
0.455
0.322
0. 406
Period 5
Arithmetic
Mean
0.457
0. 513
0.395
0.468
0. 466
0.392
0.431
0.333
0.431
0.333
0. 516
0.587
0.491
0.498
0. 504
Standard
Error
0.036
0.063
0.043
0.070
0. 105
0.040
0.097
0. 057
0.071
0.248
0. 056
0.080
0. 057
0. 112
0. 123
-------
TABLE 130. SUMMARY OF MERCURY CONCENTRATIONS IN URINE
Mercury Cone.
in Urine
ng/l
Human Subjects
Males
Females
Age
Total
0-6
7-18
19-45
46+
Total
0-6
7-18
19-45
46 +
Total
0-6
7-18
19-45
46 +
No. of
Subjects
225
45
58
90
32
106
20
34
37
15
119
25
24
53
17
Sampling
Geometric
Mean
3. 70
2. 78
4. 77
3.92
2.98
3.90
3. 03
4. 50
4. 35
3. 04
3. 53
2.60
5. 17
3.64
2.92
Period 1
Arithmetic
Mean
5. 10
4. 02
6. 04
5. 27
4. 42
5. 24
4. 32
5. 45
5. 82
4. 53
4. 98
3.78
6.88
4.89
4.32
Standard
Error
0. 25
0. 47
0. 57
0. 38
0. 59
0. 34
0. 74
0. 55
0. 61
00. 92
0. 36
0.60
1. 11
0. 49
0. 79
No. of
Subjects
190
40
47
75
28
91
18
27
33
13
99
22
20
42
15
Sampling
Geometric
Mean
1. 35
1. 00
1. 35
1.83
0. 91
1. 37
1. 09
1.36
1. 93
0. 08
1.33
0. 93
1. 35
1.76
1. 02
Period 5
Arithmetic
Mean
1.81
1.24
1.73
Z.39
1. 22
1.79
1. 28
1.72
2.45
0. 95
1.83
1. 20
1.74
2. 34
1. 45
Standard
Error
0. 11
0. 15
0. 17
0. 20
0. 23
0. 15
0. 16
0. 22
0. 31
0. 18
0. 16
0. 25
0. 29
0. 27
0. 40
-------
TABLE 131. SUMMARY OF COPPER AND ZINC CONCENTRATIONS IN BLOOD
Copper Cone, in Blood yg/100 ml
(Sampling Period 1)
Zinc Cone, in Blood yg/100 ml
(Sampling Period 1)
Age
Human Subjects
Total
0-6
7-18
19-45
46 +
(O
O Males
*" Total
0-6
7-18
19-45
46+
Females
Total
0-6
7-18
19-45
46 +
No. of
Subjects
Z07
28
57
90
32
102
14
34
39
15
105
14
23
51
17
Geometric
Mean
100.6
110. 7
99. 5
98.8
99. 2
98.0
105.3
102. 5
93.6
93.2
103.2
116.3
95.3
103. 0
104. 8
Arithmetic
Mean
103. 0
112. 1
101. 1
101. 9
101. 5
99.6
106. 1
104.0
94. 8
96.0
106. 3
118. 2
96.9
107.3
106. 5
Standard
Error
1. 51
3. 57
2.28
2.62
3.75
1.67
3.47
2. 82
2. 34
5.79
2.47
5.93
3.71
4. 12
4.71
No. of
Subjects
207
28
57
90
32
102
14
34
39
15
105
14
23
51
17
Geometric
Mean
564. 9
518. 5
540. 9
593. 1
573. 9
565. 5
533.8
522. 4
607.6
593. 0
564. 3
503. 6
569. 5
582. 2
557. 6
Arithmetic
Mean
574. 7
532. 8
551.8
600. 1
580. 5
573.6
543.3
531. 2
611.3
599.8
575.8
522.3
582.3
591. 7
563. 5
Standard
Error
7. 25
23.63
14. 66
9.68
15.68
9.32
28. 47
16.84
10. 79
23. 52
11. 11
38.64
25.63
14. 95
20.75
-------
TABLE 132. SUMMARY OF COPPER AND ZINC CONCENTRATIONS IN FECES
Copper Cone, in Feces
Wet Weight
(Sampling Period 1)
Zinc Cone, in Feces
Wet Weight
(Sampling Period 1)
Age
Human Subjects
Total
0-6
7-18
19-45
46 +
Males
Total
0-6
7-18
19-45
46+
Females
Total
0-6
7-18
19-45
46 +
No. of
Subjects
208
41
54
82
31
98
16
32
36
14
110
25
22
46
17
Geometric
Mean
15.46
14.63
17. 08
15. 54
13.79
15.37
13.64
17.43
15.73
12. 47
15. 54
15.31
16.59
15.39
14.98
Arithmetic
Mean
17. 17
15. 89
18.51
17. 50
15.67
17. 05
15. 14
19.34
17. 18
13.69
17. 28
16.38
17. 30
17. 75
17. 31
Standard
Error
0. 53
1. 02
1. 03
0. 89
1. 40
0. 80
1. 93
1. 54
1.26
1.67
0.70
1. 15
1. 16
1. 25
2. 11
No. of
Subjects
208
41
54
82
31
98
16
32
36
14
110
25
22
46
17
Geometric
Mean
95. 6 .
87. 4
102.9
99. 3
85.6
93.0
70.8
110. 2
99. 2
73. 0
98.0
99.9
93.3
99.4
97.6
Arithmetic
Mean
108. 0
116.9
111.6
106. 7
93.2
101.6
74.2
120. 0
106. 2
79.0
113.6
144. 2
99.3
107. 1
104. 8
Standard
Error
6. 5
30.3
5.9
4.6
6.8
4.3
6.4
8. 2
6.3
8.8
11.6
49. 1
7.9
6.7
9.3
-------
TABLE 133. SUMMARY OF COPPER AND ZINC CONCENTRATIONS IN HAIR
Copper Cone, in Hair (ig/g
Dry Weight
(Sampling Period 1}
Zinc Cone, in Hair |ig/g
Dry Weight
(Sampling Period 1)
Age
Human Subjects
Total
0-6
7-18
19-45
46+
to
Ox Mates
Total
0-6
7-18
19-45
46+
Females
Total
0-6
7-18
19-45
46 +
No. of
Subiects
228
49
58
89
32
107
22
33
37
15
121
27
25
52
17
Geometric
Mean
29. 0
22. 1
36.7
32. 5
21. 1
25.1
19.7
32.3
26,4
18.4
33.0
24.3
43.2
37.7
23.9
Arithmetic
Mean
37. 7
27. 2
44. 4
42.6
28.0
31.7
24.9
39.3
32.8
22. 2
43.0
29.0
51. 1
49.5
33. 1
Standard
Error
2. 1
3. 2
3. 8
3. 8
5.6
2.5
5. 0
4.2
4.8
4.3
3. 2
4. 1
6.8
5.3
9.9
No. of
Subjects
229
49
59
89
32
108
22
34
37
15
121
27
25
52
17
Geometric
Mean
170. 5
129. 5
163. 5
189.8
208. 1
147.6
98.7
154. 4
163. 2
188. 1
193.9
161.6
176. 9
211.3
227. 5
Arithmetic
Mean
289. 7
315.6
245.3
259.9
414.6
223.7
120.0
276.6
187. 1
346.5
348. 5
474. 9
202.7
311.7
474.7
Standard
Error
40.6
131.7
70. 4
36.3
131. 4
46. 5
20.6
121. 1
24. 1
182.8
64.3
235. 9
27.2
59.0
191. 5
-------
TABLE 134. SUMMARY OF COPPER AND ZINC CONCENTRATIONS IN URINE
Copper Cone, in Urine Mg/l
(Sampling Period 1)
Zinc Cone, in Urine yg/l
(Sampling Period 1)
Age
Human Subjects
Total
0-6
7-18
19-45
46+
to
O Males
^ Total
0-6
7-18
19-45
46+
Females
Total
0-6
7-18
.19-45
46 +
No. of
Subjects
225
45
58
90
32
106
20
34
37
15
119
25
24
53
17
Geometric
Mean
10. 02
11. 81
9.69 .
10. 12
8.22
9.89
11.98
9. 20
9.73
9.44
10. 13
11.67
10.43
10.40
7.28
Arithmetic
Mean
11. 14
13. 23
10. 76
11.19
8.78
10.89
12.97
10.41
10. 71
9.68
11.37
13. 45
11. 26
11. 52
7.99
Standard
Error
0. 34
1. 05
0.55
0.51
0. 52
0. 42
1. 14
0.72
0.72
0. 59
0.53
1.67
0. 85
0.71
0.79
No. of
Subjects
225
45
58
90
32
106
20
34
37
15
119
25
24
53
17
Geometric
Mean
423
476
452
400
372
429
426
444
445
363
418
520
462
372
381
Arithmetic
Mean
498
627
501
458
4Z2
478
458
498
495
418
516
762
506
432
426
Standard
Error
33.9
154. 6
32.3
24. 5
34. 7
21.6
40.6
44. 0
36.3
52. 5
61.2
276.0
47.9
32.8 -
47.6
-------
TABLE 135. SUMMARY OF BLOOD HEMATOCRITS
Hematocrits
in Blood % I
No. of
Age Subjects
Human Subjects
Total 203
0-6 27
7-18 56
19-45 88
46+ 32
Males
Total 98
0-6 13
7-18 33
19-45 37
K> - 46+15
O
oo
Females
Total 105
0-6 14
7-18 23
19-45 51
46+ 17
Sampling
Geometric
Mean
40. 4
37.3
38.9
41.7
42.3
41.5
37.4
39.6
43.5
44.5
39.4
37. 1
38.0
40.4
40. 5
Period 1
Arithmetic
Mean
40.6
37.3
39.0
41.8
42.5
41.7
37.5
39.7
43.7
44.6
39.5
37. 1
38.0
40.5
40.6
Standard
Error
0.27
0. 50
0.38
0.40
0.63
0.42
0.80
0.50
0.64
0.72
0.31
0.61
0. 52
0.43
0.73
No. of
Subjects
199
26
56
88
29
93
11
32
37
13
106
15
24
51
16
Sampling
Geometric
Mean
41.4
38. 1
40.0
42.9
42.7
42.6
38.9
40.2
45,5
44.1
40.3
37.6
39.8
41.1
41.5
Period 2
Arithmetic
Mean
41. 5
38.2
40. 1
43.0
42.8
42.8
38.9
40.3
45.5
44.2
40.4
37.6
39.9
41. 1
41.6
Standard
Error
0.24
0.37
0.39
0.34
0.55
0.40
0.44
0.60
0.41
0.86
0.24
0.52
0.43
0.30
0.59
No. of
Subjects
168
35
40
68
25
79
13
23
30
13
89
22
17
38
12
Sampling
Geometric
Mean
41.3
38. 1
40.8
42.7
43.1
43.0
38.6
41.4
45.7
44.6
39.9
37.8
40.0
40.5
41.5
Period 5
Arithmetic
Mean
41. 5
38.2
40.9
42.9
43.2
43.2
38.6
41.6
45.8
44.7
40.0
37.9
40. 1
40.6
41.6
Standard
Error
0.29
0.37
0.48
0.47
0.67
0.44
0.42
0.71
0. 51
0.95
0.31
0. 54
0.52
0. 50
0.70
-------
Some recurrent age and sex patterns were obtained in comparing the trace metal data for a single period.
The adult age groups tended to have higher metal concentrations in their blood specimens than did the child
age groups. However, the children generally had higher metal concentrations in their feces and hair
specimens. Females tended to have higher metal concentrations in feces than did the males, while the males
generally had higher metal levels in their urine.
The changes in metal concentration for a specimen between sampling periods are much more pro-
nounced, however, than are the age and sex patterns. For example, the mean levels of mercury in the blood
and urine specimens decreased in Period 5 (October 1976) to less than half of the levels observed in Period 1
(October 1974). The mean lead levels in the feces and urine samples also decreased substantially in Period 5, to
just over half of the Period 1 levels.
The two-sample t-test was conducted to test the null hypothesis of equal means in periods 1 and 5 against
the two-sided alternative. Prior analyses have shown clinical specimen metal data to be lognormally
distributed/49'50^ The relatively large standard errors and the relatively large differences between the cor-
responding geometric and arithmetic means displayed in Tables 119 - 134 suggest each of these clinical
specimen trace metal data sets follow a lognormal distribution rather than a normal distribution. Thus, the
two-sample t-tests for differences were performed on natural log transferred data (i.e., assuming a lognormal
distribution) for the three trace metals (cadmium, lead, and mercury) in the four clinical specimens (blood,
feces, hair, and urine). The hematocrits data appears normally distributed; thus its Period 1 vs. Period 5 com-
parison was made on the untransformed data.
The results of these tests for differences in mean metal level between Periods 1 and 5 are presented in
Table 136. The t-statistic has been converted to its probability level P and interpreted with regard to the direc-
tion and significance of the detected change. There were significant decreases from Period 1 (baseline) to
Period 5 (operational) for cadmium in feces (P = .001) and hair (P = .02), for lead in feces (P « .0001),
and for mercury in blood (P « .0001), hair (P < .0001); and urine (P « .0001). The only significant in-
crease from Period 1 to Period 5 was for hematocrits.
TABLE 136. CLINICAL SPECIMEN TRACE METAL CONCENTRATION AND HEMATOCRITS PATTERNS
Trace Metal
Cadmium:
Lead:
Mercury:
Hematocrits
Significant Change in Geometric Mean from Baseline Period to Operational Period
Blood
Possible increase
(P = 0.08)
No difference
Large decrease
(P « 0.0001)
Increase
(P = 0.02)
Feces
Decrease
(P = 0.001)
Large decrease
(P < 0.0001)
No difference
Haii
Decrease
(P = 0.02)
Possible decrease
(P = 0.18)
Large decrease
(P< 0.0001)
Urine
Possible decrease
(P = 0.11)
Large decrease
(P < 0.0001)
Large decrease
(P « 0.0001)
Note: Probability levels P are for a two-sided test.
Since the detected changes in the operational period clinical specimen trace metal concentrations were all
decreases, any sewage treatment plant contribution to these levels has obviously been countered and over-
whelmed by the factors responsible for the decrease' The identity of these factors is unknown. However, the
209
-------
facts that the different periods samples were stored for differing lengths of time, analyzed at different times,
and usually analyzed by different operators, leads to speculation that the differences in levels may be due to
analytical factors. In summary, the clinical specimen trace metal data provides no evidence of any adverse
sewage treatment plant health effect.
HUMAN SUBJECT CURRENT HEALTH STATUS
The Current Health Status (CHS) Questionnaire completed by each participant during each of his sampl-
ing periods was presented in Figure 5 in Section 5. This questionnaire was patterned after the Household
Health Survey Questionnaire form, both in the illnesses, diseases, and symptoms investigated, and in the time
periods covered. The basis of response and the time period covered for each health category in the CHS
questionnaire can be summarized as:
Health Category Basis of Response Time Period Covered
Chronic illnesses Receiving medication At present
Diseases Diagnosis Past year
Symptoms Experienced Past three months
At present
The dates of the sampling periods and the participant groups (original and/or Lexington Green) pro-
viding their current health status were:
Period 1 Oct., 1974 Original participants
Period 2 Feb., 1975 Original participants
Periods Oct., 1975 Lexington Green participants
Period 4 Feb., 1976 Original participants
Periods Oct., 1976 Original and Lexington Green
participants
The Egan plant commenced operations in December, 1975. Subsequently, the participants were in-
structed in reporting symptoms to substitute Jan. 1—Feb. 20 for "past three months" in Period 4 and the
period from March through October for "past three months" in Period 5.
There were 187 of the original participants who completed the Current Health Status questionnaire in all
four of the periods they were sampled (Periods 1, 2, 4, and 5). Thirty-two of the participants from the
Lexington Green Apartments completed both of their CHS questionnaires (Periods 3 and 5). The positive
responses to each chronic illness, disease, and symptom were tabulated for these 187 original participants and
32 Lexington Green participants. The reported incidence of each illness, disease, and symptom has been
displayed for the corresponding baseline and operational periods, and accumulated over the relevant baseline-
operational comparisons.
The number of participants who were presently receiving medication for each chronic illness is presented
in Table 137. Taking medication for asthma-hay fever decreased in incidence, particularly in the Fall. Chronic
sinusitis and bronchitis and hypertension had substantial increases in incidence, with the increase in chronic
sinusitis and bronchitis occurring in the Fall.
The incidence of disease diagnosis during the previous year is shown in Table 138. The February data on
the original participants can not be accumulated with their October data in the total valid comparisons
because their one year periods of reporting overlap. The increases reported in the February survey for
pneumonia and skin disease may be important, because they may reflect the effect of recent exposure before
immunity to a sewage treatment pathogen has developed. A decrease in the reported incidence of anemia
occurred in the October surveys.
210
-------
TABLE 137. CURRENT HEALTH STATUS DATA SUMMARY: INCIDENCE OF
"PRESENTLY RECEIVING MEDICATION FOR CHRONIC ILLNESSES"
Original Original Lexington Total Valid
Participants Participants Green Comparisons -
(October) (February) Participants All Participants
Per. 1 Per. 5 Per. 2 Per. 4 Per. 3 Per. 5 Baseline Operational
Participants Responding 187 187 187 187 32 32 406 406
CHRONIC ILLNESSES:
Tuberculosis 0000 0000
Malignancies 0000 0 00 0
Asthma-Hay Fever 13 8 3 5 5 2 21 15
Diabetes 1212 1135
Heart Conditions 4111 1163
Hypertension 2635 2 27 13
Chronic Sinusitis 6 11 5 4 0 2 11 17
& Bronchitis
Arthritis s 4427 309 11
Rheumatism
Rheumatic Heart 1000 0010
Disease
Thyroid Disease 7477 0 0 14 11
Liver Disease 0000 0000
Kidney Disease 1000 0010
TABLE 138. CURRENT HEALTH STATUS DATA SUMMARY: INCIDENCE OF
"DISEASES DIAGNOSED DURING PAST YEAR"
Original . Original Lexington Total Valid
Participants Participants Green Comparisons*-
(October) (February) * Participants All Participants
Per. 1 Per. 5 Per. 2 Per. 4 Per. 3 Per. 5 Baseline Operational
Participants Responding 187 187 187 187 32 32 219 219
DISEASES:
Polio 0000 0000
Infectious 0000 0000
Jaundice
Pneumonia 59490059
Worms 3010 0030
Skin Disease 5738 0057
Pleurisy 3214 0032
Spinal 0100 0 00 1
Meningitis
Influenza 21 19 32 31 15 22 24
Croup 4 4 7 3 0 0.4 4
Sleeping Sickness 0001 0000
Anemia 4122 3172
Dysentery 3021 0000
Empyema 0000 0000
Substantial
Change
in
Incidence
-6
+•6
+6
Substantial
Change
in
Incidence
( + 5)
( + 5)
-5
* The February data ray not represent valid comparisons because their tune periods overlap the October time periods, and
because of the brief period of Egan Plant operation.
211
-------
The incidence of symptoms reported in each Current Health Status survey is presented in Table 139 for
the previous "three month" period. The nominal "three month" period was less than 2 months in Period 4
and more than 7 months in Period 5. Thus, no valid comparisons can be made from Table 139 on the
incidence of symptoms experienced during the previous "three months."
The incidence of symptoms being presently experienced during each survey is presented in Table 140.
Accumulating incidence over all valid comparisons, there was a decrease in the incidence of diarrhea, but
increases in the incidence of severe headache not relieved by aspirin, severe pain in bones and joints with high
fever, colicky pains in abdomen, and shortness of breath. There also was an increase in the incidence of sore
throats and colds in the February surveys, which may reflect recent exposure to sewage treatment pathogens,
before immunity has developed.
A statistical analysis of this Current Health Status data was performed to determine from the self- and
seasonally-paired data whether there were any significant changes from the baseline to the operational period
in the incidence of any of the reported illnesses, diseases, or symptoms. These paired comparison health data
have the same nature as the paired isolations of bacteria, parasites, and viruses in the clinical specimens.
Thus, the same statistical procedure was used. Sign tests, with two-sided levels of significance, were used to
test the null hypothesis of no difference in incidence of the illnesses, diseases, and symptoms between the valid
paired comparisons for the baseline and operational survey periods. Pairs for which occurrence was reported
in both the baseline and the operational period were excluded from the analysis. When the number of pairs
showing a change was greater than 10, the normal distribution approximation to the binomial expansion was
used to calculate the probability level. A change of five in the incidence for the most extreme case (a 0-5 split)
yields a two-sided probability level of .062. Thus only illnesses, diseases, and symptoms with at least a dif-
ference of 5 in incidence between the baseline and operational periods were considered for analysis by the sign
test, since all the other changes would be found insignificant.
All of the illnesses, diseases, and symptoms with a baseline vs. operational change in incidence of at least
five over all valid comparisons are indicated in the last column of Tables 137-140. A change of incidence of at
least five in the February surveys is also indicated in parentheses in this column. The sign test analysis of these
incidence changes is presented in Table 141.
As Table 141 shows, no significant changes were detected in the incidence of any chronic illness, disease,
or symptom at the 5% significance level. There were, however, a number of changes having a sufficiently
small probability level to possibly reflect a real difference in health status. Those health measures having a
possibly significant increase in incidence over both operation periods were hypertension, severe pain in bones
and joints with high fever, colicky pains in abdomen, and shortness of breath. Skin disease incidence also
showed a possibly significant increase in the February operational period. The decrease in the incidence of
anemia and diarrhea was possibly significant.
It should be noted that the incidence of most of the health measures in the Current Health Status survey
was so low that only very substantial changes in health status could be detected in our participants. However,
there were 18 valid comparison health measures presented in Tables 137-140 that had a combined baseline and
operational incidence of ten or more. Even if there were no difference in health status between the baseline
and operational periods, one would still have expected one significant change at the 5% significant level just
due to random variation. Thus the Current Health Status survey does not, by itself, provide any evidence of a
possible sewage treatment plant health hazard.
212
-------
TABLE 139. CURRENT HEALTH STATUS DATA SUMMARY: INCIDENCE OF
"SYMPTOMS EXPERIENCED DURING PAST THREE MONTHS"*
Original
Participants
• Per. 1 Per. 5
Participants Responding
SYMPTCMS:
Severe Headache not
Relieved by Aspuin
Severe Dizziness
Severe Pain in Bones
and Joints with High Fever
Severe Weight loss
Severe Night Sweats
Henorrhagic Rash
Canker Sores Around ftouth
Yellow Eyeballs
Sore Inroat
Cough
Cold
Skin Rash: Face
Skin Rash; Arras I, Legs
Skin Rash: Body
Diarrhea
Bloody Diarrhea
Bloody urine
Burning on Urination
Nausea
Vomiting
Yellow Skin
Pain in Chest on Deep Breathing
Weakness of Arms or Legs
Cough up Blood
Stiff Neck with Fever
Stiff Neck with Rash
General Weakness
Droning FJT-
Fever above 103° F
Severe Trouble with Teeth
Colicky Pains in Abdomen
Brown Urine
Shortness of Breath
Convulsions
Unconsciousness (not due to
blow on head
167
7
5
3
1
2
0
9
0
48
58
69
4
5
6
34
1
0
1
22
19
0
4
4
0
1
0
7
2
9
2
5
0
4
0
0
187
20
7
3
2
4
0
13
0
83
90
100
7
13
9
37
0
0
4
30
26
0
8
13
1
4
1
29
5
11
1
10
0
8
0
2
Original
Participants
187
16
3
2
0
2
1
14
0
63
94
100
g
7
5
44
1
0
2
31
31
0
5
6
0
3
1
14
3
12
3
10
0
4
0
1
187
17
5
9
1
5
2
13
0
71
83
94
6
9
6
31
0
0
1
26
25
0
4
8
1
5
0
20
1
16
4
13
0
10
0
0
LBxington
Green
Participants*
PerTT Per. 5
32
3
0
1
0
1
0
5
0
8
4
10
1
0
0
5
0
0
1
0
0
0
0
0
0
1
0
2
0
1
1
1
0
0
0
0
32
6
1
1
0
1
0
5
1
15
16
22
3
3
3
8
0
0
1
9
5
0
3
3
0
0
0
8
2
1
1
6
0
1
0
0
*Because the period "past three^rcnths" was reduced to 7 weeks for Period 4 and extended to over 7 months for
Period 5, there are.no valid oonparisons for statistical analysis.
213
-------
TABLE 140. CURRENT HEALTH STATUS DATA SUMMARY: INCIDENCE OF
"SYMPTOMS EXPERIENCING AT PRESENT"
Original
Participant*
(October)
Per.T Per. 5
Participants Responding
SYMPTOMS:
Severe Headache not
Relieved by Aspirin
Severe Dizziness
Severe Pain in Rnryg
t Joints with High
Fever
Severe Weight Loss
Severe Night Sweats
Hemorrnagic Rash
Canker Sores Around
(touth :
Yellow Eyeballs
Sore Throat
Cough
Cold
Skin Rash: Face
Skin Rash: Arms & Legs
Skin Rash; Body
Diarrhea
Bloody Diarrhea
Bloody Urine
Burning on Urination
Nausea
vomiting
Yellow Skin
Pain in Chest on
Deep Breathing
Weakness of Arms
or Legs
Cough up Blood
Stiff Neclt with
Fever
Stiff Neck with
RasE
General Weatoess
Draining Ear
Fever above 103°F
Severe Trouble
with Teeth
Colicky Pains in
Abdoren
Brown Urine
Shortness of Breath
Convulsions
Unconsciousness (not
due to blow on head)
187
2
1
0
0
0
0
1
0
9
18
17
2
2
3
5
0
0
0
1
0
0
1
0
0
0
0
2
0
0
2
1
0
1
0
0
187
2
0
0
0
0
0
2
0
1
11
11
1
3
0
0
0
0
0
1
1
0
1
1
0
0
0
1
0
3
0
3
0
2
0
0
Original
Participants
(February)
PerTT Per. 4
187
3
2
0
0
2
0
6
0
7
29
26
2
5
2
3
0
0
1
2
0
1
4
3
0
1
0
2
2
0
1
1
0
1
0
0
187
5
1
5
0
3
1
1
0
13
32
33
1
4
2
2
0
0
1
1
0
0
0
4
0
1
0
6
0
1
1
5
0
6
0
0
Lexington
Green
Participants
PerT Per. 5
32
0
0
0
0
0
0
0
0
4
3
5
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
32
3
1
0
0
0
0
1
0
2
3
6
1
0
0
1
0
0
1
1
0
0
1
0
0
0
0
0
0
0
0
0
0
0
0
0
Total Valid
Conparlflons-
All Participants
Baseline Operational
406 406
5
3
0
0
2
0
7
1
20
50
48
4
7
5
8
0
0
1
3
0
1
5
3
0
1
0
4
2
0
3
2
0
2
0
0
10
2
5
0
3
1 '
4
0
16
46
50
3
7
2
3
0
0
2
3
1
0
2
5
0
1
0
7
0
4
' 1
a
0
B
0
0
Substantial
Change
in
Incidence
+5
+5
(+6)
(+7)
-5
+6
Because the period "past three months* was reduced to 7 weeks for Period 4 and extended to over 7 months for Period 5,
there are no valid oanparisons for statistical analysis.
214
-------
TABLE 141. CURRENT HEALTH STATUS STATISTICAL ANALYSIS:
SIGN TEST ON INCREASES AND DECREASES IN INCIDENCE
CHRONIC ILLNESSES;
(presently receiving medication)
Asthma-Hay Fever
Hypertension
Chronic Sinusitis and Bronchitis
DISEASES:
(during past year)
Pneumonia (February)
Skin Disease (February)
Anemia
Paired Incidence Changes
Baseline Operational
Only Only
16
2
8
10
8
14
Probability
Level*
P
.33
.11
.29
.27
.18
.06
Sign Test Result
Interpretation
of
Significance
No Change
Possible Increase
No Change
No Change
Possible Increase
Possible Decrease
SYMPTOMS:
(experiencing at present)
Severe Headache not
Relieved by Aspirin
Severe Pain in Bones and
Joints with High Fever
Sore Throat (February)
Cold (February)
Diarrhea
Colicky Pains in Abdomen
Shortness of Breath
5
19
7
2
1
11
26
2
8
7
.27 No Change
.06 Possible Increase
.21 No Change
.37 No Change
.18 Possible Decrease
.11 Possible Increase
.07 Possible Increase
215
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SECTION 7
DISCUSSION
This study has examined numerous public health hazards which might possibly be associated with the
operation of sewage treatment plants. Many findings relevant to sewage treatment health effects were
obtained in Section 6, "Results" from statistical analyses and observations of the study data. These findings
are summarized by study area (Results Section) in table 142. Each finding has also been classified in Table
142 with regard to its health hazard implication for the sewage treatment plant studied. The classification
system employed is:
+ + Strong indication of a potential health hazard
+ Indication of potential health hazard
+ (?) Uncertain indication of potential health hazard
? Health implication is uncertain
— (?) Uncertain indication of no health hazard
— Indication of no health hazard
Strong indication of no health hazard
(blank) Irrelevant to health implications
The health importance, statistical significance, and methodological difficulties associated with each finding
were considered in obtaining its hazard classification.
The Egan plant was a reasonable study site. The aeration basin contained fairly high levels of many
pathogenic bacteria (Pseudomonas, fecal Streptococci, Klebsiella, Mycobacteria, Staphylococcus, and Pro-
videncia). However, Salmonella and Shigella, "the most important wastewater-borne enteric pathogenic
bacteria encountered in the United States,"51 were not detected. Coliphage and enteroviruses were found in
nearly every sample. The analytical methods used indicate that most of the enteroviruses were polioviruses
and that they were found primarily in the solids fraction of the mixed liquor. The first stage aeration basin
contained high concentrations of cadmium, lead, and mercury in Period 5.
The air sampling shows that the Egan plant's aeration basins are a localized source of indicator
organisms in the air (standard plate count, total coliform, and fecal coliform). The occasional presence of
hardy pathogenic microorganisms (Pseudomonas, fecal Streptococci, Klebsiella, and a single enterovirus
(typed as poliovirus III) in the downwind samples suggests that the aeration basin is also an aerosol source of
these pathogens. Most of the microbiological particles were in the respirable range. The first stage aeration
basin also appears to be a source of aerosolized mercury. However, there were no significant differences
between any residential upwind and residential downwind levels fc300 m). The levels of microbiological or
chemical agents of the air, water, and soil samples in the residential areas, were not distinguishable from the
background levels.
The levels of lead in the soil and of total coliforms and mercury in the water were significantly higher
after the sewage treatment plant was in operation, but in each case the increase did not primarily occur in the
samples taken close to the sewage treatment plant. No pathogenic bacteria or viruses were isolated in any soil
or water sample. Again, no contribution of the sewage treatment plant to microbiological or chemical levels
in the soil and water in the plant vicinity could be detected.
216
-------
TABLE 142. SUMMARY AND CLASSIFICATION OF FINDINGS REGARDING HEALTH
IMPLICATIONS OF A SEWAGE TREATMENT FACILITY
Hazard
Classifi-
Study Area of Results Findings Relevant to Sewage Treatment Health Effects cation
A. ENVIRONMENTAL MONITORING
1. Aeration Basin Samples
a. Microbial characterization
9 The levels of Klebsiella, Mycobacteria, Staphylococcus, and Providencia + (?)
were fairly high relative to the microbiological indicator levels.
b. Microbiological data
• Salmonella, Shigella, and Proteus (pathogenic bacteria) were not detected.
• Enteroviruses (presumably poliovirus) were nearly always found, but pre- ?
dominantly in the solids fraction of the sample.
• Pseudomonas, fecal streptococci, and coliphage were consistently found. + (?)
• The first stage aeration basin had much higher coliform levels than did
the second stage.
c. Trace metal data
• The first stage aeration basin contained much higher concentrations of cad-
mium, lead, and mercury in October than did the second stage in February.
• There was little daily variation in the levels of the trace metals.
2. Air Samples
a. Microbiological data
• Klebsiella was occasionally detected in the aerosol downwind of the aeration + (? )
basin, as far as 300 m.
• An enterovirus plaque was detected in one aerosol sample, at 300 m down- + (?)
wind from the aeration basin.
• Coliphage and fecal streptococci were found infrequently in the downwind — (?)
aerosol.
(continued)
-------
Study Area of Results
TABLE 142. (continued)
Findings Relevant to Sewage Treatment Health Effects
to
co
• Most microbiological particles in the aerosol emanating from the aera-
tion basin are in the respirable range (primarily 3.3 to 7. 0 microns in
diameter).
• The aeration basins are a significant source of total coliforms, fecal
coliforms, and standard plate count. However, there is no significant
difference between the residential upwind and residential downwind
(>300 m) levels of total coliform, fecal coliform, or standard plate
count.
• There is no significant difference between the upwind and close downwind
levels of Pseudomonas.
b. Trace metal data
• The first stage aeration basin was a borderline significant source of aero-
solized mercury. However, there is no difference in the mercury levels
between the residential upwind and residential downwind locations.
• There is no significant difference between the upwind and close downwind
levels of cadmium or lead.
3. Soil Samples
a. Microbiological data
• No pathogenic bacteria (particularly Streptococcus-beta, Staphylococcus
aureus, Salmonella, Shigella, and Klebsiella pneumoniae) were isolated
in any soil sample.
• No viruses were isolated via plaques or cytopathogenie effect.
• The standard plate count was significantly lower in the operational peri-
ods than in the baseline periods.
b. Trace metal data
• The lead levels were significantly higher in the operational periods than
in the baseline periods, but the increase occurred in the distant samples
rather than in the samples taken close to the sewage treatment plant.
• There were no significant differences between the baseline and the opera-
tional levels of cadmium, mercury, copper, and zinc.
Hazard
Classifi-
cation
(continued)
-------
Study Area pf Results
TABLE 142. (continued)
Findings Relevant to Sewage Treatment Health Effects
to
MD
4. Water Samples
a. Microbiological data
• No pathogenic bacteria (particularly Salmonella, Shigella, and Vibrio)
or fecal coliforms were found in any water sample.
• No viruses were isolated via plaques or cytopathogenic effect.
• The MPN total coliform level was significantly higher in the operational
periods than in the baseline periods, but the increase was unrelated to
distance from the sewage treatment plant.
b. Trace metal data
• The mercury levels were significantly higher in the operational periods
than in the baseline .periods, but the increase was unrelated to distance
from the sewage treatment plant.
• There was no significant difference between the baseline and the opera-
tional levels of lead.
B. HOUSEHOLD HEALTH SURVEY
Hazard
Classifi-
cation
(continued)
1. Household and Personal Characteristics
• The operational survey households had higher occupational status, fewer
bedrooms, and more air conditioning than the baseline households. The
two surveys did not differ in racial composition, head of household edu-
cation, or household size.
• The operational survey had a more balanced distance and directional dis-
tribution about the sewage treatment plant.
• The operational survey households were located closer to expressways,
multilane highways, and industrial operations. Their length of residence
was longer.
• Fewer young children were surveyed in the operational period, both over
all and within 2. 0 kilometers of the sewage treatment plant. The effect o
this age pattern must be taken into account for those health parameters fc
which a potential health effect is observed, and for which incidence is re-
lated to age.
-------
Study Area of Results
TABLE 142. (continued)
Findings Relevant to Sewage Treatment Health Effects
to
t-o
o
2. Analysis of Health Survey Illness, Disease, and Symptom Incidence
• There were significant increases in incidence in the operational period
close to (within 2 km) the sewage treatment plant (with a significant
survey x distance interaction), occurring mainly in the downwind direc-
tions, for the symptoms of nausea, vomiting, and general weakness.
Influenza displayed the same pattern, but with only a slight increase
close to the plant.
• There were patterns of significantly increased incidence close to the
sewage treatment plant (without a significant interaction), occurring
mainly in the downwind directions, for skin disease and the symptoms
of diarrhea and pain in chest on deep breathing.
• There was a pattern of elevated incidence close to the sewage treatment
plant (with a significant interaction) for dysentery.
• There were significant decreases in incidence in the operational period
survey (with a significant interaction), especially close to the sewage
treatment plant, for worms and sore throat.
• There were significant operational survey decreases in incidence (with-
out a significant interaction), also occurring close to the sewage treat-
ment plant, for chronic sinusitis and bronchitis and for fever above
103°F.
• The significantly increased incidence of skin disease, diarrhea, nausea,
vomiting, general weakness, and pain in chest on deep breathing, and
the elevated incidence of influenza, close downwind from the sewage
treatment plant cannot be attributed to age confounding. The increased
close-in incidence of these health parameters tended to occur over all
age categories. Furthermore, influenza, diarrhea, nausea, and vomit-
ing generally occur more frequently among children, but there were fewer
children within 2 kilometers of the sewage treatment plant in the opera-
tional survey, so that a decreased close-in incidence would have been an-
ticipated from the surveyed age distribution.
Hazard
Classifi-
cation
(continued)
-------
Study Area of Results
TABLE 142. (continued)
Findings Relevant to Sewage Treatment Health Effects
All of the diseases and symptoms displaying significantly increased in-
cidence (skin disease, diarrhea, nausea, vomiting, general weakness,
and pain in chest on deep breathing) or elevated incidence (influenza)
close downwind from the sewage treatment plant during the operational
survey are considered, from a medical standpoint, to be likely or possi-
bly related to the presence of a sewage treatment plant.
All of the health parameters (chronic sinusitis and bronchitis, worms,
sore throat, and fever above 103°F) having significantly decreased inci-
dence close to the sewage treatment plant are also considered, from a
medical standpoint, to be likely or possibly related to the presence of a
sewage treatment plant.
to
to
C. HUMAN SUBJECT CHARACTERISTICS
Only the additional human subjects recruited from the Lexington Green
Apartments lived within 1 km of the sewage treatment plant. These ad-
ditional subjects differed from the original human subjects by consisting
of. more females and fewer school age children.
A majority (51%) of the human subjects expected to spend no more than
one hour per day more than 2 miles from their homes.
Hazard
Classifi-
cation
D. CLINICAL SPECIMEN MICROBIOLOGY
1.
Bacterial Isolations
a. Feces samples
There was a significant decrease in Proteus isolations in the operational
periods.
There were no significant differences in the isolations of Pseudomonas or
Salmonella between the baseline and operational periods.
(continued)
-------
Study Area of Results
TABLE 142. (continued)
Findings Relevant to Sewage Treatment Health Effects
KJ
tO
I-O
b. Throat swabs
* There were very significant increases in the isolation of Streptococcus -
beta and Staphylococcus aureus in the operational periods. Regression
analysis suggests that the increases are unrelated to the level of the
participant's sewage treatment plant exposure.
* Isolations of Streptococcus-alpha and Streptococcus-gamma increased
significantly for the close-in (400m-600m) Lexington Green subjects in
the operational period. Isolations in the operational periods decreased
significantly for Streptococcus-alpha and remained unchanged for Strep-
tococcus-gamma for the more distant original subjects. Regression
yields some evidence that these isolations may be related to sewage
treatment plant exposure.
c. Sputum samples
• No Mycobacterium tuberculosis was isolated in any sputum sample.
Parasite Isolations
• There was a nearly significant increase in parasitic incidence in feces in
the operational periods. However, improvements in sample collection
methods and the upstream location of all five parasitic subjects' residences
suggest the increase may be unrelated to the sewage treatment plant.
Viral Isolations
a. Feces samples
• The only viruses isolated were Picornavlruses, which include polio-
viruses, Coxsackieviruses, and Echoviruses. There was a significant
increase in Picornavirus isolations in the operational periods. The in-
crease does not appear to be related to sewage treatment plant exposure.
b. Throat swabs
• No viruses were isolated in any of the throat swab samples.
Hazard
Classifi-
cation
(continued)
-------
TABLE 142. (continued)
Hazard
Classifi-
Study Area of Results Findings Relevant to Sev.-age Treatment Health Effects cation
4. Viral Antibody Titers
a. Tests on all human subjects
• There was a significant increase in the poliovirus I titer obtained by
serum neutralization, which appears to be unrelated to sewage treat-
ment plant exposure.
• Tests for six types of polio, Coxsackie, and Echo viruses showed no
difference in titer between the baseline and operational period samples.
There was a significant decrease in Echovirus 12 titer by hemagglutina-
tion inhibition.
b. Additional tests on 100 human subjects
• The Echovirus 29 titer obtained by hemagglutination inhibition increased
,0 significantly in the operational period, with the increase showing a
w slight exposure relationship, but only among the original participants.
• There were no significant differences in titer level for the other 22 viral
antibody tests.
E. CLINICAL SPECIMEN TRACE METALS
The following clinical specimen trace metal concentration patterns were
found for the operational period relative to the baseline period:
1. Cadmium
• Significantly decreased cadmium levels in the feces and in the hair.
Possibly decreased urine levels and possibly increased blood levels.
2. Lead
• Very significantly lower lead levels in the feces and urine. A possible
decrease in hair lead levels. No change in blood lead levels.
(continued)
-------
TABLE 142. (continued)
Study Area of Results
Findings Relevant to Sewage Treatment Health Effects
3. Mercury
• Very significantly lower mercury levels in the blood, hair, and urine.
No change in the fecal mercury level.
4. Hematocrits
• A significant increase in the hematocrits percentage of the blood.
F. HUMAN SUBJECT CURRENT HEALTH STATUS
• There were no significant changes from the baseline period to the opera-
tional period in the incidence among the human subjects of taking medica-
tion for any chronic illness, of diagnosis of any disease, or of experiencing
any symptom.
• There was a possibly significant increase in the incidence of hypertension,
skin disease in February, severe pain in bones and joints with high fever,
colicky pains in abdomen, and shortness of breath.
• There was a possibly significant decrease in the incidence of anemia and
diarrhea.
Hazard
Classific-
cation
-------
The household health survey indicated that increased incidence of skin disease and the symptoms of
nausea, vomiting, general weakness, diarrhea, and pain in chest on deep breathing occurred close to the
sewage treatment plant and predominantly in the downwind direction after it was in operation. The facts that
each of these diseases and symptoms could be caused by pathogens found or presumed to exist in the aeration
basin and that their incidence is not attributable to changes in the surveyed age distribution suggest the sewage
treatment plant as a possible cause. However, a "normal" sporadic outbreak of these diseases and sympto-
matology in the schools or communities just north and south of the plant is also a reasonable explanation.
Since the survey relied on the respondent's memory of prior diseases and symptoms within the family, its
evidence is non-medical and possibly subjective. Also, the possibility that dissatisfaction with the treatment
plant stimulated the memory of some nearby residents cannot be dismissed.
The possibility for biased responses from participants was considered during recruitment. All prospective
participants were told the objectives of the study. Each was informed that they were to be a part of the study
and none were told that they were a part of a group close to the plant or far from the plant. It, therefore,
seems improbable that biased responses would be obtained from participants which are related to distance
and in some cases direction from the plant.
The incidence with which some bacteria, parasites, and viruses were isolated in the clinical specimens
increased in the operation periods. Isolations of beta hemolytic Streptococci and Staphylococcus aureus in the
throat swabs increased very significantly. There were significantly increased isolations of Picornaviruses in
feces and, for the closest subjects, of alpha and gamma hemolytic Streptococci in their throat swabs. The
increase in the incidence of parasitic protozoa in the feces was nearly significant. Regression analysis of the
isolation increases with respect to distance, direction, hours home, and air conditioning level indicated that
most of these isolation increases appeared unrelated to the subject's anticipated exposure to the sewage treat-
ment plant's aerosol. The exceptions were alpha and gamma hemolytic streptococci in the throat swabs, for
which the increase did bear some relation to presumed aerosol exposure, although these are not of practical
health significance.
It is surprising that no adenoviruses or reoviruses were isolated in over 800 feces samples, and that no
viruses at all were isolated in more than 900 throat swabs. It is not known if there were deficiencies in the
methods of collection, shipping, or the holding time prior to analysis. In retrospect, our practice of storing all
of the frozen samples from a sampling period and shipping them as a batch to the analytical laboratory was
probably unwise. It overburdened the laboratory upon receipt and delayed the sample analysis. However, in
sampling Periods 2, 4 and 5, approximately two positive control samples each of feces and throat swabs were
seeded with poliovirus in the field, shipped with the regular samples, and analyzed blind; in each case the
poliovirus was isolated.
Viral serology detected significant operational period titer increases for poliovirus I and Echovirus 29,
with the Echovirus 29 titer increases showing a slight relationship to sewage treatment plant exposure only
among the original participants. There was a significant decrease in Echovirus 12 titer and no change in titer
for 28 other viral antibodies.
The environmental monitoring did not show that the sewage treatment plant was a source of trace metals
for ambient air in the residential area. It is not surprising that levels of these metals in the participants (hair,
blood, urine and feces) were not related to the sewage treatment plant. The levels of lead and cadmium seen in
the participants are comparable to other studies*46-50) of middle class suburban populations without occupa-
tional exposures to these metals. Differences in levels of lead in the air and soil and in participants might be
explained by their proximity to a freeway in the study area.
Although not an objective of this study, the trace metal data on more than 200 participants will provide
important information regarding levels present in suburban populations in the United States. Air data for
225
-------
each trace metal are available for each participant's neighborhood over a two-year period. Detailed question-
naire information is available for each participant and data for hair, urine, blood and feces for the trace
metals are available for the same participants sampled during a two-year span.
If the aeration basin, air, soil, and water sampling were to establish the sewage treatment plant as an
aerosol source of a particular pathogen or metal, and if there were corresponding appropriate patterns of its
elevated levels in the clinical specimens, and of the increased incidence of its resulting symptoms and diseases,
then this corroboration would strongly implicate sewage treatment as a public health hazard. However, the
environmental monitoring, clinical specimen, and health survey findings are too isolated to provide cor-
roboration. The symptoms and diseases with possible plant-related patterns have too many potential causes to
warrant any association with the pathogens and trace metals found to have an exposure-like pattern.
The experimental design employed in this study was to compare baseline and operational levels during
each of two seasons. When significant increases occurred, their occurrence was related to measures of sewage
treatment plant exposure such as distance and direction to infer the relationship to exposure. This is a sen-
sitive design for identifying health hazards and inferring whether the sewage treatment plant may be their
source. However, the findings obtained in this study, when considered overall, did not detect a public health
hazard for persons living beyond 400 meters from a well-operated sewage treatment plant. Greater sensitivity
is required before an epidemiology study can be expected to obtain firm evidence regarding the potential
health hazard closer than 400 meters from a sewage treatment plant. Methods to achieve greater sensitivity
include the recruitment of health survey respondents and human subjects living considerably closer to the
sewage treatment plant than in this study, the use of daily health diaries, and improving the collection, ship-
ping, and analytical methodologies in such areas as pathogen isolation.
The most useful sources of health effects information in this study have been microbiological
environmental monitoring, the household health survey, and viral serology. These areas should form the basis
of future epidemiological investigations of sewage treatment plant health hazards.
226
-------
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Medicine & Surgery, 34, 2, 130 (Feb. 1965).
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5. Ladd, F. C., "Airborne Bacteria from Liquid Waste Treatment Units". M.S. thesis, Oklahoma State
Univ., Stillwater (1966).
6. Randall, C. W., and Ledbetter, J. O., "Bacterial Air Pollution from Activated Sludge Units".
Amer. Ind. Hyg. Assn. Jour., 27, 6, 506 (Nov-Dec. 1966).
7. Kenline, P. A., and Scarpino, P. V., "Bacterial Air Pollution from Sewage Plants". Amer. Ind.
Hyg. Assn. Jour., 33, 5, 346 (May 1972).
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9. Pereira, M. R., and Benjaminson, M. A., "Microbial Air Pollutants Disseminated by a Sewage
Treatment Plant". Paper presented at Symp. on Aerobiology, Science and Man in the Americas,
Amer. Acad. for Advancement of Sci., Mexico City (July 3, 1973).
10. Adams, A. P., and Spendlove, J. C., "Coliform Aerosols Emitted by Sewage Treatment Plants".
Science, 169, 3951, 1218 (September 18, 1970).
11. Sorber, C. A., et al., Problem Definition Study: "Evaluation of Health and Hygiene Aspects of
Land Disposal of Wastewater at Military Installations". U.S. Army Medical Environ. Eng. Res.
Unit Report (No.)73-02, Army Medical Res. and Development Command, Edgewood Arsenal,
Maryland (August 1972).
12. Jensen, K. E., "Presence and Destruction of Tubercule Bacilli in Sewage". Bull. World Health Org.,
10,2,171(1954).
13. Sorber, C. A., et al., "An Assessment of a Potential Virus Hazard Associated with Spray Irrigation
of Domestic Wastewaters". Paper presented at meeting on Virus Survival in Water and Wastewater
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14. Ledbetter, J. O., et al., "Health Hazards from Wastewater Treatment Practices". Environ. Lett., 4,
225 (1973).
15. Putnam, R., and Paulus, H. J., presented at Amer. Industrial Hygiene meeting, San Francisco
(1972).
16. Dowling, H. F., "Airborne Infection - the Past and the Future". Bacterial. Rev., 30, 3, 485 (Sept.
1966).
17. Personal Communication from Mr. Tim McAloon, plant operator, John E. Egan Water
Reclamation Plant.
18. Scaringelli, F. P., Puzak, J. C., Bennett, B. I., and Denny, R. L., "Determination of Total Mercury
in Air by Charcoal Adsorption and Ultraviolet Spectrophotometry". Analytical Chemistry, 46, 2
(1977).
19. Long, S. J., Scott, D. R., and Thompson, R. J., "Atomic Absorption Determination of Elemental
Mercury Collected from Ambient Air on Silver Wool". Analytical Chemistry, 45, 13 (1973).
20. Gardner, D., "A Rapid Method for the Determination of Mercury in Air by Flameless Atomic
Absorption Spectrometry". Analy. ChimActa, 82, pp 321-327 (1976).
21. Johnson, D. E., et al., Evaluation of Health Effects Associated with the Application of Wastewater
to Land, Phase II, Pre-Fair Report. Southwest Research Institute report to U.S. Army Medical
Research and Development Command, October 1976.
22. Taras, M. J., Greenberg, A. E., Hoak, R. D., and Rand, M. C., Standard Methods for the
Examination of Water and Wastewater, 13th Edition. American Public Health Association,
Washington, D.C. (1971).
23. Rand, M. D., Greenberg, A. E., Taras, M. J., and Franson, M. A., Standard Methods for the
Examination of Water and Wastewater, 14th Edition. American Public Health Association,
Washington, D.C. (1975).
24. Lenette, E. H., Spauling, E. H., and Truant, J. P., Manual of Clinical Microbiology, 2nd Edition.
American Society for Microbiology, Washington, D.C. (1974).
25. Dubos, R. J., and Hirsch, J. G. (ed.), Bacterial and Mycotic Infections of Man. 4th Edition, J. P.
Lippincott, Philadelphia (1965).
26. Lenette, E. H., and Schmidt, N. (ed.), Diagnostic Procedures, Viral and Rickettsial Infections.
American Public Health Association, Washington, D.C. (1969).
27. Horsfall, F. L., Jr., and Tamm, I. (ed.), Viral and Rickettsial Infections of Man, 4th Edition, J. P.
Lippincott, Philadelphia (1965).
28. Methods for Chemical Analysis of Water and Wastes. Methods Development and Quality Assurance
Research Laboratory, National Environmental Research Center, Environmental Protection Agency,
Cincinnati, Ohio (1974).
29. Analytical Methods for Atomic Absorption Spectrophotometry. Perkin Elmer Corp., Norwalk,
Conn., revised March 1973, pp BC-23.
228
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30. Thompson, R. J., Morgan, G. B., and Purdue, L. J., Air Quality Instrumentation, Volume 1, J. W.
Scales (ed.) Instrument Society of America, Pittsburg, Pennsylvania (1972).
31. Wroblewski, S. C., Spinier, T. M., and Harrison, P. R., Journal of the Air Pollution Control
Association, 24, 8, (1974).
32. Smith, R. G., and Windom, H. R., The Marine Resource Center, Savannah, Georgia, unpublished
manuscript (1972).
33. Chester, R., and Hughes, J. J., Chem. Geol., 2, pp 249-62.
34. Hwang, J. Y., Ullucci, P. A., Mokeler, C. L, Analytical Chemistry, 45, 4, (1973).
35. Hauser, T. R., Hinner, T. A., and Kent, J. L., Analytical Chemistry, 44, 11, 1819(1972).
36. Mitchell, D. G., Ryan, F. J., andAldous, K. M., Atomic Absorption Newsletter, Vol. 11, No. 6, 120
(1972).
37. Hatch, W. R., Ott, W. L., Analytical Chemistry, 40, 14 (1968).
38. Skare, I., Analyst, Vol. 97, pp 148-55 (1972).
39. Rantala, R.T.T., Loring, D. H., Atomic Absorption Newsletter, Vol. 12, No. 4, 97-98 (1973).
40. Hammer, D. I., Finklea, J. F., Hendricks, R. H., Hinners, T. A., Riggan, W. B., Shy, C. M., Trace
Substances in Environmental Health-V, DD. Hemphill (Ed.). University of Missouri, Columbia,
(1972).
41. Harrison, W. W., Yuracheck, J. P., Benson, C. A., Clinical Chimca Acta, 23, pp 83-91, (1969).
42. Kubasik, N. P., Volosin, M. T., Clinical Chemistry, Vol. 19, No. 9, 954 (1973).
43. Parker, M. M., Humoller, F. L., Mahler, D. J., Clinical Chemistry, 13, No. 1, pp 40-48 (1967).
44. Johnson, D. E., et al. Evaluation of Health Effects Associated with the Application of Wastewater
to Land, Phase II report to U. S. Army Medical R&D Command (in preparation).
45. Brownlee, K. A., Statistical Theory and Methodology in Science and Engineering, 2nd Edition, New
York: John Wiley and Sons, Inc., (1960).
46. Johnson, D. E., Prevost, R. J., Tillery, J. B., Kimball, K. T., and Hosenfeld, J. M., Epidemiologic
Study of the Effect of A utomobile Traffic on the Blood Levels of Persons in Selected Age Groups,
Final report to Environmental Protection Agency for EPA Contract 68-02-2227 (March 1977).
47. Snedecor, G. W., and Cochran, W. G., Statistical Methods, 6th Edition, Ames, Iowa: Iowa State
University Press (1967).
48. Kirk, R. E., Experimental Design: Procedures for the Behavioral Sciences. Belmont,
California/Brooks/Cole Publishing Co. (1968).
49. Johnson, D. E., Prevost, R. J., Tillery, J. B., Camann, D. E., and Hosenfeld, J. M., Baseline Levels
of Platinum and Palladium in Human Tissue, EPA-600/1-76-019 (March 1976).
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50. Johnson, D. E., Prevost, R. J., Tillery, J. B., and Thomas, R. E., The Distribution of Cadmium and
Other Metals in Human Tissue, Southwest Research Institute report to Environmental Protection
Agency, Contract 68-02-1725, (September 1977).
51. Cooper, Robert C., "Health Considerations in Use of Tertiary Effluents". Journal of the
Environmental Engineering Division, Proc. of Amer. Soc. of Civil. Eng., Vol. 103, No. EE1,
(Feb. 1977).
230
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APPENDIX A
JUSTIFICATION OF SURVEY QUESTIONNAIRE FORMS
Clearance requests and justification for each of the three questionnaire forms developed and used in this
study are presented in this Appendix. Each justification is preceded by an OMB Form 83, Clearance Request
and Notice of Action.
231
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STANDARD FORM HO. 83
OPF'CE OF MANAGE MEN'
AND BUDGET
CLEARANCE REQl'FST AND NOTICE OF ACTION
(Under Federal Reports Act and Bunau of the Budget Circular No. A-40, as amended)
FOR O.M.B. USE
IMPORTANT - Submit the required number or copies or SF-83, together
with the material lex which approval is request*"* to:
R£AD INSTRUCTIONS BEFORE COMPLETING FORM
CLEARANCE OFFICER
OFFICE OF MANAGEMENT AND BUDGET
WASHINGTON, D.C. 20503
PART A - REQUEST BY FEDERAL AGENCY FOR CLEARANCE
* Items marked with asterisk may be omitted for preliminary plans or recordkeeping requirements
1. SEND "NOTICE OF ACTION" TO: Name and mailing address
2. Bureau and division or office originating
request
3. Name(s), title(s), and telephone numbers ol
person's) who can best answer questions
regard ing request.
FORM OR
DOCUMENT
IDENTIFI-
CATION
4. Title or form or document submitted
HOUSEHOLD HEALTH SURVEY
*5. Agency Form Number(s|
. Type of form or document
i fj] Application
2 rj Program evaluation
3 Q Other management
report
4|~~1 Statistical survey
or report
s| | Preliminary plan
or contract
6 fj Recordkeeping
requirement
7 n Other -Specify
7. Current (or former) O.M.B. clearance •
Number
Expiration date
8. Requested expiration date
9. Type of request
I (x]New
2 [ ) Revision
xtension
(No change)
4Q Reinstatement
*10. Frequency of use
i Q Single time s CD Quarterly
2 S On occasion s CD Semi-annually
3 CD Weekly 7 Q Annually
lonthly B n Other
-------
JUSTIFICATION OF HOUSEHOLD HEALTH SURVEY
1. Supporting Statement Justification
(1) This new questionnaire is required to obtain information on a
population living near a sewage treatment plant to determine whether or
not certain diseases are associated with the operation of sewage treatment
plants. No forms of this type are currently available within the Environ-
mental Protection Agency, or known from other agencies that meet the
requirements of this design form. This questionnaire form is to be
utilized on the EPA Contract No. 68-02-1746, SwRI Project entitled "Health
Implications of Sewage Treatment Facilities. " It has been shown that the
operation of an activated sludge treatment plant does produce certain types
of pathogens and trace metals which may be associated with the introduc-
tion of disease and residues of these trace metals in a nearby population.
This questionnaire form would provide necessary information to guide the
Environmental Protection Agency in its assessment of potential health
effects associated with the operation of an activated sludge treatment plant.
The questionnaire form will solicit information from 1, 000 households to
cover all individuals living in the household in regard to age, sex, chronic
diseases, infectious diseases, and certain types of symptoms that may be
related to these infectious diseases. The plan is to survey approximately
1, 000 households prior to the operation of a new activated sludge treatment
plant and to follow up the survey with an additional 1, 000 households once
233
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the plant has become operational. This project will be conducted as a
contract with Southwest Research Institute under the technical direction
of Dr. Donald E. Johnson. The attached statement from Southwest Research
Institute provides more details on the plans for this survey.
(2) The data collected using this survey instrument and the follow-up
statistical analysis will be utilized by the Environmental Protection Agency
to assess the potential health hazards associated with the operation of an
activated sludge treatment plant.
(3) Some preliminary data have been published in the literature
relative to the operation of a sewage treatment plant near populated areas,
however, this in-depth survey is required to obtain more definitive
information.
2. Description of Survey Plan
(1) The survey design will be aimed at determining the disease incidence
of a population living within 5 kilometers of an activated sludge treatment
plant in populations within the United States. The survey will be directed
at one typical sewage treatment plant and will provide an estimation of
the types of data found. It is estimated that total population living within
5-kilometer radii of all activated sludge sewage treatment plants within the
United States is over one million.
(2) The survey is designed to contact and survey 1, 000 households
selected at random within a 5-kilometer radius of a typical activated sludge
234
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sewage treatment plant. The plant selected will have at least a population
ten times greater than the 1, 000 households needed. The survey designed
provides for examination of 1, 000 households prior to the operation of
the activated sewage plant and a follow up study at least six months after
the sewage plant has become operational. The intent of the study is to
compare the incidence of certain diseases before and during the operation
of a new activated sludge sewage treatment plant. The households will be
contacted by a personal interview survey team in which the individual
answering the door will be asked to answer questions for himself and
all other members of the household. The completed questionnaires will
be forwarded to the survey team and then appropriate statistical analyses
will be made following completion of the two surveys of 1, 000 households
to correlate differences in incidence of diseases. A pre-test of the
questionnaire has been conducted. This pre-test was conducted with
five representative individuals from the staff at Southwest Research
Institute, and no difficulties were encountered. The question of non-
responders for this particular survey does not appear to represent a
problem since the burden for the individuals at households to respond to
this is minimal and number of non-responders is expected to be very
low. Preliminary contact with representatives of the community to be
surveyed indicates that this type of response will be obtained.
(3) The statistical design of the project will come primarily from
Mr. David Camann with Southwest Research Institute. The Environmental
235
-------
Protection Agency's statistician, ,
has reviewed this protocol.
(4) Name of the Contractor: Southwest Research Institute. Contrac-
tor's role: the primary role is to collect information and provide a
final report to include statistical evaluation of data and conclusions
regarding the potential health effects related to the operation of an
activated sludge sewage treatment plant with a disease survey.
Southwest Research Institute guarantees confidentiality of the
collected data to the subjects surveyed. No direct reference to the
collected data using the subject's name or address will be made. The
compilation of subject's name and address will be maintained in a con-
fidential file and will not be directly related to any of the collected data.
Each subject surveyed will be assigned a code number and the coded
number with the individual's name will be maintained only in the files
of Dr. Donald E. Johnson, principal investigator, at Southwest Research
Institute.
3. Time Schedule for Data Collection and Publication
(1) The contract is scheduled for two-year completion, including
monthly reports followed by a comprehensive final report to
be submitted on completion of the two-year study. Attached is a time
schedule for the entire project. It is estimated that the elapsed time
236
-------
between the completion of data collection and the issuance of the first
published reports will be approximately one year.
4. Consultations Outside the Agency
Detailed consultations were conducted with Dr. Joe E. Ledbetter,
Professor of Civil Engineering, University of Texas at Austin, regarding
the overall design of the survey. Dr. Ledbetter has conducted some of the
previous studies connecting certain disease incidence with the operation
of activated sludge plants. Mr. Lee Riser, L. L. Hiser and Associates,
New Braunfels, Texas, has also been consulted with regard to the conduct
of the survey. Mr. Riser's expertise is primarily in the area of design
and operation of sewage treatment facilities and his expertise was
valuable in determining a typical activated sludge sewage treatment plant for
use in the survey.
Consultation has also been made with the Metropolitan Sanitary
District of Greater Chicago with Mr. Bart Lyman, its general superin-
tendent. Additional consultations have been made with Mr. Raymond
Rimkus, Chief of Maintenance and Operations, and Dr. Lue-Hing, Director
of Research and Development of the Metropolitan Sanitary District of
Greater Chicago for their cooperation and assistance in the conduct of
the surveys near one of their activated sludge sewage treatment plants.
237
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5. Estimation of Respondent Reporting Burden
The estimation of respondent reporting burden for the Household
Health Survey is ten minutes. This estimate is based on a preliminary
pre-test survey of randomly selected Southwest Research Institute employees,
and should be the amount of time necessary for even a large household to
complete.
6. Sensitive Questions
Questions 5, 6, and 7 ask questions regarding recent state of
health in the subjects of the household being surveyed. This information
is essential in order to provide an assessment of the health effects
associated with a sewage treatment plant. Questions 9, 10, 11, 12,
13 and 14 provide information about how long the individuals within the
household have lived within the area to be studied which is relevant to
the exposure time to the potential health hazards within the area. Other
parts of these questions refer to some of the socio-economic background
necessary to make valid statistical comparisons with other areas.
238
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STANDARD FORM HO. 33
OFFICE O* MANAGE ME-,
AND BUO&C T
CLEARANCE REQI'PST AND NOTICE OF ACTION
(Under Federal Reports Act and Bunau of the Budget Circular No. A-40, as amended)
FOR O.M.B. USE
IMPORTANT - Submit the required nu'ber of copies ol SF-B3. torether
with the rrjtenal (of wmcn approval is requested to:
READ INSTRUCTIOHS BEFORE COMPLETING FORM
CLEARANCE OFFICER
OFFICE OF MANAGEMENT AND BUDGET
WASHINGTON, D.C. 20503
PART A - REQUEST BY FEDERAL AGENCY FOR CLEARANCE
* Items marked with asterisk may be omitted for preliminary plans or recorakeeping requirements
1. SEND "NOTICE OF ACTION" TO: Name and mailing addtess
2. Bureau and division or office originating
request
3. Name(s), title(s), and telephone numbers of
person(s) who can best answer questions
regarding request.
FORM OR
DOCUMENT
IDENTIFI-
CATION
4. Title of form or document submitted
HEALTH SURVEY PARTICIPANT QUESTIONAII
*5. Agency Form Number(s)
IE
6. Type of form or document
i PJ Application
2 PJ Program evaluation
3 PJ Other management
report
4 | | Statistical survey
or report
51 | Preliminary plan
or contract
6 PJ Recordkeeping
requirement
7 PJ Other - Specify—
7. Current (or former) O.M.B. clearance •
Number
Expiration date
8. Requested expiration date
9. Type of request
i (x) New
2 | | Revision
3 P] Extension
(No change)
41 | Reinstatement
*10. Frequency of use
i PJ Single time 5rjQua'terlv
2 [x] On occasion 6 PJ Semi-annually
3 Q Weekly 7nAnnuallv
4 pj Monthly en °th«r f5M''ns"uc"°n5)
11. Related forms or documents (Give O.M.S number. Enclose in
parentheses any to be rep/oce<{)
12. Catalog of Federal Domestic Assistance program number (i( applicable)
COLLECTION
AND
RESPONDENTS
*13a. Collection method
{Check as many OS apply)
1 Q Mail
2 Q9 Personal interview
3 PJ Other - Describe —,
*13b. Collected by -
4 pj Agency
5 | | Contractor
£ L7I Other - Describe—,
14a. Type of respondents involved
(Check preo'cxTinonr one)
1 (5? Individuals or households
2 PJ Business firms (non-farm)
3 PJ Farms
4 CD Government agencies
5 PJ Other - Describe —,
*15. Summary of estimated
respondent burden
». Estimated number of
respondents
14b. Brief description of respondents
(i.e., "households in 50 largest
SMSA's; "reliil grocery stores")
b. If sample, approximate
number in universe
c. Reports filed annually by
each respondent (item 10)
d. Total annual responses (a X c)
e. Estimated average number of
iftan-hours required per response
I. Estimated TOTAL MAN-HOURS
of respondent burden (d X el
Number
300
over
1 million
once
300
1/2
150
AUTHORITY
AND COMF1-
DEHTIALITY
'llja.ls te '. form -
I [~2\ V:luntary?
2 "X R auired to obtain benefit?
3 Q M; ndatory? Ore srofuie -
i *16b. DOTS your a.-.ency pied,-.e
I confidentiality?
I Yes
2P/1 No
CONSULTA-
TIONS OUT-
SIDE AGENCY
17. In developing the report form or otlrer documents, were
consultations held with individuals or organirations
outside your agency?
i (XI Yes - If "yes," identify persons and describe outcome m
r-. ,, SUPPORTING STATEMENT. (See instmciions)
m NO
CERTIFICATION BY AUTHORIZED OFFICIALS SUBMITTING REQUEST - We certify th.1t the form or other document submitted for approval is
necessary for the proper perlormance of this agency's functions, that the information requested is not available from any other source, to the best
of our knowledge, and that the request is consistent with applicable O.M.B. and agency policy directives. Signature and title ol:
Approving official for agency
Daie
Agency clearance officer or other agency official
Date
239
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JUSTIFICATION OF HEALTH SURVEY PARTICIPANT QUESTIONNAIRE
1. Supporting Statement Justification
(1) This new questionnaire is required to obtain information on a
population living near a sewage treatment plant to determine whether or
not certain diseases are associated with the operation of sewage treatment
plants. No forms of this type are currently available within the Environ-
mental Protection Agency, or known from other agencies that meet the
requirements of this design form. This questionnaire form is to be utilized
on the EPA Contract No. 68-02-1746, SwRI Project entitled "Health Implica-
tions of Sewage Treatment Facilities. " It has been shown that the opera-
tion of an activated sludge treatment plant does produce certain types
of pathogens and trace metals which may be associated with the introduc-
tion of disease and residues of these trace metals in a nearby population.
This questionnaire form would provide necessary information to guide the
Environmental Protection Agency in its assessment of potential health
effects associated with the operation of an activated sludge sewage
treatment plant. The questionnaire form will solicit health-related
information from 200-240 individuals living in the study area in regard to
age, sex, chronic diseases, infectious diseases, and certain types of
symptoms that may be related to these infectious diseases. The plan
is to survey and sample approximately 200-240 individuals prior to the
operation of a new activated sludge treatment plant and to follow up the
240
-------
survey with an additional 200-240 individuals once the plant has become
operational. This project will be conducted as a contract with Southwest
Research Institute under the technical direction of Dr. Donald E. Johnson.
The attached statement from Southwest Research Institute provides more
details on the plans for this survey.
(2) The data collected using this survey instrument and the follow-up
statistical analysis will be utilized by the Environmental Protection Agency
to assess the potential health hazards associated with the operation of an
activated sludge sewage treatment plant.
(3) Some preliminary data have been published in the literature
relative to the operation of a sewage treatment plant near populated areas,
however, this in-depth survey is required to obtain more definitive
information.
2. Description of Survey Plan
(1) The survey design will be aimed at determining the disease
incidence of a population living within 5 kilometers of an activated sludge
sewage treatment plant in populations within the United States. The survey
will be directed at one typical sewage treatment plant and will provide an
estimation of the types of data found. It is estimated that total population
living within 5-kilometer radii of all activated sludge sewage treatment plants
within the United States is over one million.
241
-------
(2) The survey is designed to initially contact and survey 1, 000
households selected at random within a 5-kilometer radius of a typical
activated sludge sewage treatment plant. The plant selected will have at
least a population ten times greater than the 1, 000 households needed.
The survey designed provides for examination of 1, 000 households prior
to the operation of the activated sewage plant and a follow up study at
least six months after the sewage plant has become operational. The
intent of the study is to compare the incidence of certain diseases before
and during the operation of a new activated sludge sewage treatment
plant. The households will be contacted by a personal interview survey
team in which the individual answering the door will be asked to answer
questions forwarded to the survey team and then appropriate statistical
analyses will be made following completion of the two surveys of 1, 000
households to correlate differences in incidence of diseases. If the sub-
ject responds in the negative to question 10 of the Hous ehold.Health
survey: "Do you plan to move during the coming year?" and in the
affirmative to question 15: "Would you participate in a health survey
as a paid volunteer?" he will be issued a Health Survey Participant
Questionnaire. It is anticipated that between 200 and 240 subjects who
respond to this questionnaire will be chosen, on the basis of age and
location categories, to participate as paid volunteers. Each subject will
be sent a letter announcing that they have been selected for the survey;
242
-------
a time and place will be specified as to when they should report for a
first meeting and for sampling dates.
A pre-test of the questionnaire has been conducted. This pre-test
was conducted with five representative individuals from the staff of Southwest
\
\
Research Institute, and no difficulties were encountered. The question of
non-responders for this particular survey does not appear to represent a
problem since the burden for the individuals at households to respond
to this is minimal and number of non-responders is expected to be very
low. Preliminary contact with representatives of the community to be
surveyed indicates that this type of response will be obtained.
(3) The statistical design of the project will come primarily from
Mr. David Camann with Southwest Research Institute. The Environmental
Protection Agency's statistician, ,
has reviewed this protocol.
(4) Name of the Contractor: Southwest Research Institute, Contrac-
tor's role: the primary role is to collect information and provide a final
report to include statistical evaluation of data and conclusions regarding
the potential health effects related to the operation of an activated sludge
sewage treatment plant with a disease survey.
Southwest Research Institute guarantees confidentiality of the
collected data to the subjects surveyed. No direct reference to the
collected data using the subject's name or address will be made. The
243
-------
compilation of subject's name and address will be maintained in a
confidential file and will not be directly related to any of the collected
data. Each subject surveyed will be assigned a code number and the
coded number with the individual's name will be maintained only in the
files of Dr. Donald E. Johnson, principal investigator, at Southwest
Research Institute.
3. Time Schedule for Data Collection and Publication
(1) The contract is scheduled for two-year completion, including
monthly reports followed by a comprehensive final report to be
submitted on completion of the two-year study. Attached is a time schedule
for the entire project. It is estimated that the elapsed time between the
completion of data collection and the issuance of the first published reports
will be approximately one year.
4. Consultations Outside the Agency
Detailed consultations were conducted with Dr. Joe E. Ledbetter,
Professor of Civil Engineering, University of Texas at Austin, regarding
the overall design of the survey. Dr. Ledbetter has conducted some
of the previous studies connecting certain disease incidence with the
operation of activated sludge plants. Mr. Lee Hiser, L. L. Hiser and
Associates, New Braunfels, Texas, has also been consulted with regard
244
-------
to the conduct of the survey. Mr. Hisers expertise is primarily in the
area of design and operation of sewage treatment facilities and his expertise
was valuable in determining a typical activated sludge treatment plant for
use in the survey.
Consultation has also been made with the Metropolitan Sanitary
District of Greater Chicago with Mr. Bart Lyman, its general superinten-
dent. Additional consultations have been made with Mr. Raymond Rimkus,
Chief of Maintenance and Operations, and Dr. Lue-Hing, Director of
Research and Development of the Metropolitan Sanitary District of
Greater Chicago for their cooperation and assistance in the conduct of
the surveys near one of their activated sludge sewage treatment plants.
5. Estimation of Respondent Reporting Burden
The estimation of respondent reporting burden for the Health Survey
Participant Questionnaire is six minutes. This estimate is based upon a
preliminary pre-test survey of randomly selected Southwest Research Institute
employees, and should be the amount of time necessary even if all
members of a given household choose to participate in the health survey
and sampling program.
6. Sensitive Questions
Questions 5, 6, 7, 8, 18 and 19 provide background information on
individuals that will be utilized for correlation with other studies and for
245
-------
making estimates with regard to general populations at risk. Question 9
asks the subject about how many hours of the day he normally spends
more than 2 miles from his home. This question is required to obtain
some estimate of the total time the individuals spend within the study area.
Question 10 asks what is the natural color of respondent's hair. This
information is needed because it has been shown that the natural color of
an individual's hair will affect the levels of certain trace metals found in
hair. Since it is a part of the study to analyze trace metals in hair samples,
this information is essential. Questions 11 and 12 ask about the cigarette
smoking status of the individual. This information is needed for corre-
lation with several parameters, but one includes trace metal informa-
tion since it has been shown that cigarette smoking does provide a different
profile of certain metals such as cadmium in cigarette smokers versus
non-smokers. Questions 13, 14 and 15 will provide an assessment of
how long the subjects have lived within the survey area and how long they
plan to continue living in the area. This information is needed both for
purposes of estimating the length of exposure of subjects to the sewage
treatment plant and to establish a stable survey population over the 18-
month period required. Question 16 asks about chronic illnesses and
certain medications that the subject may be taking with regard to these
illnesses. It is necessary to obtain information regarding chronic illnesses
in order to provide an assessment of the type of population being surveyed
246
-------
with comparison to populations in other areas of the country. Additionally,
it is necessary since some trace metal surveys have shown that certain
chronic diseases significantly affect trace metal levels in different tissues.
Also, the medication taken by the individual has been shown to affect
these levels, thus this type of correlation would be of importance.
Question 17 relates to home air conditioning. Since it is anticipated that
the major avenue of transport of certain pathogens and trace metals from
the sewage treatment plant to the population is via ambient air, it is
important to know whether or not the subject has air conditioning, since
this will affect this relationship.
247
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STANDARD FORM HO. 83
rriCC OF MANAGEMENT
MD BUDGET
CLEARANCE REQI'FST AND NOTICE OF ACTION
(Under Federal Reports Act and Bureau ol the Budget Circular No. A-40, as amended)
FOR O.M.B. USE
IMPORTANT - Submit the required number of copies of SF-83. together
with the material for which approval is requested to:
READ INSTRUCTIONS BEFORE COMPLETING FORM
CLEARANCE OFFICER
OFFICE OF MANAGEMENT AND BUDGET
WASHINGTON, D.C. 20503
PART A - REQUEST BY FEDERAL AGENCY FOR CLEARANCE
* Items marked with asterisk may be omitted lor preliminary plans or recordkeeping requirements
I. SEND "NOTICE OF ACTION" TO: Name and mailing address
2. Bureau and division or office originating
request
3. Name(s), title(s), and telephone numbers of
p«rson(5) who can best answer questions
regarding request.
FORM OR
DOCUMENT
IDENTIFI-
CATION
4. Title of form or document submitted
CURRENT HEALTH STATUS QUESTIONNAIRE
*5. Agency Form Number(s)
6. Type of form or document
i | | Application
2 O Program evaluation
3 Q] Other management
report
41 | Statistical survey
or report
5|| Preliminary plan
or contract
g Q] Recordkeeping
requirement
7 Q Other - Specify—
7. Current (or former) O.M.B. clearance •
Number
Expiration date
8. Requested expiration date
9. Type of request
i |x) New
2 | | Revision
3 | | Extension
(No change)
einstatement
'10. Frequency of use
I CD Single time 5 Q Quarterly
2 (3 On occasion s Q Semi-annually
3 Q Weekly 7 Q Annually
11. Related forms or documents (Give O.M.a number. Enc/ose in
porenrrieses any ro
12. Catalog of Federal Domestic Assistance program number (if applicable)
COLLECTION
AND
RESPONDENTS
*13a. Collection method
(Oiecfc os many as
2 Q3 Personal interview
3 Q Other - Describe _
*13b. Collected by -
4 Q Agency
5 [J Contractor
6 CIJ Other - Describe -
14a. Type of respondents involved
(Check predtyninonr one)
i [x* Individuils or households
2 Q] Busines-. firms (non-farm)
3 O Farrm
4 Q Government agencies
5 Q Other - Describe —r
14b. Brief description of respondents
(i.e., "households in 50 largest
SMSA's; "retiil grocery stores")
'15. Summary of estimated
respondent burden
a. Estimated number of
respondents
b. If sample, approximate
number in universe
c. Reports filed annually by
each respondent (item 10)
d. Total annual responses (a X c)
e. Estimated average number of
man-hours required per response
t. Estimated TOTAL MAN-HOURS
of respondent burden (d X e)
Number
150
over
1 million,
600
1/2
300
AUTHORITY
AND CONFI-
DENTIALITY
*16a.ls repr-t form -
i f~l Voluntary?
2 [^5 R,-quired to obtain benefit?
3 Q] M; ndatory? Circ sroture
,*16b. Does >cu' .vency pledr.e
i confidentiality?
i ) S Yes 2 [7! No
CONSULTA-
TIONS OUT-
SIDE AGENCY
17. In developing the report form or other documents, were
consultations held with individuals or organirations
outside yotr agency?
\K] Yes - If "yes," identify persons and describe outcome in
r^ ., SUPPORTING STATEMENT. (See insrrucrioos)
2[JNo
CERTIFICATION BY AUTHORIZED OFFICIALS SUBMITTING REQUEST - We certify th.it tlie form or other document submitted lor approval is
necessary lor the proper perlormance of this agency's functions, that" ths information requested is not available from any other source, to the best
of our knowledge, and that the request is consistent with applicable O.M.B. and agency policy directives. Signature and title of:
Approving official for agency
Date
Agency clearance officer or other agency official
Date
248
-------
JUSTIFICATION OF CURRENT HEALTH STATUS QUESTIONNAIRE
1. Supporting Statement Justification
(1) This new questionnaire is required to obtain information on a
population living near a sewage treatment plant to determine whether or
not certain diseases are associated with the operation of sewage treatment
plants. No forms of this type are currently available within the Environ-
mental Protection Agency, or known from other agencies that meet the
requirements of this design form. This questionnaire form is to be utilized
on the EPA Contract No. 68-02-1746, SwRI Project entitled "Health Implica-
tions of Sewage Treatment Facilities. " It has been shown that the operation
of an activated sludge sewage treatment plant does produce certain types
of pathogens and trace metals which may be associated with the introduc-
tion of disease and residues of these trace metals in a nearby population.
This questionnaire form would provide necessary information to guide the
Environmental Protection Agency in its assessment of potential health
effects associated with the operation of an activated sludge sewage treat-
ment plant. The questionnaire is designed to determine the type of
medication sample volunteers may be taking and the type of diseases that
may be prevalent in the study group. It will also record the symptoms that
the subject has experienced during the previous 3 months. This informa-
tion will be used to assist in determining the incidence of disease in the
study group. This project will be conducted as a contract with Southwest
249
-------
Research Institute under the technical direction of Dr. Donald E. Johnson.
The attached statement from Southwest Research Institute provides more
details on the plans for this survey.
(2) The data collected using this survey instrument and the follow-
up statistical analyses will be utilized by the Environmental Protection
Agency to assess the potential health hazards associated with the opera-
tion of an activated sludge sewage treatment plant.
(3) Some preliminary data have been published in the literature
relative to the operation of a sewage treatment plant near populated
areas; however, this in-depth survey is required to obtain more definitive
information.
2. Description of Survey Plan
(1) The survey design will be aimed at determining health incidence
of a population living within 5 kilometers of an activated sludge sewage
treatment plant in populations within the United States. The survey will
be directed at one typical sewage treatment plant and will provide an
estimation of the types of data found. It is estimated that the total
population living within 5-kilometer radii of all activated sludge sewage
treatment plants within the United States is over one million.
(2) 200-240 subjects will be chosen from the respondents to the
Health Survey Participation Questionnaire whose participation was
250
-------
solicited from the initial Household Health Survey Questionnaire. The
Current Health Status Questionnaire will be completed by each subject
completing a sampling period. Thus, if the subject has completed the
full protocol, he will have provided four samples of urine, blood, feces,
throat swab, sputum, hair and have completed the current health status
questionnaire four times. A pre-test of the questionnaire has been
conducted. This pre-test was conducted with five representative
individuals from the staff at Southwest Research Institute, and no diffi-
culties were encountered. The question of non-responders for this
particular survey does not appear to be a problem since those individuals
asked to complete the questionnaire have already evidenced their willing-
ness to participate in a sampling program.
(3) The statistical design of the project will come primarily
from Mr. David Camann with Southwest Research Institute. The Environ-
mental Protection Agency's statistician, ,
has reviewed this protocol.
(4) Name of the Contractor: Southwest Research Institute, Contrac-
tor's role: the primary role is to collect information and provide a final
report to include statistical evaluation of data and conclusions regarding
the potential health effects related to the operation of an activated sludge
sewage treatment plant with a disease survey.
251
-------
Southwest Research Institute guarantees confidentiality of the
collected data to the subjects surveyed. No direct reference to the
collected data using the subject's name or address will be made. The
compilation of subject's name and address will be maintained in a
confidential file and will not be directly related to any of the collected
data. Each subject surveyed will be assigned a code number and the
coded number with the individual's name will be maintained only in the
files of Dr. Donald E. Johnson, principal investigator, at Southwest
Research Institute.
3. Time Schedule for Data Collection and Publication
(1) The contract is scheduled for two-year completion, including
monthly reports followed by a comprehensive final report to be submitted
on completion of the two-year study. Attached is a time schedule for the
entire project. It is estimated that the elapsed time between the com-
pletion of data collection and the issuance of the first published reports
will be approximately one year.
4. Consultations Outside the Agency
Detailed consultations were conducted with Dr. Joe E. Ledbetter,
Professor of Civil Engineering, University of Texas at Austin, regarding
the overall design of the survey. Dr. Ledbetter has conducted some of the
252
-------
previous studies connecting certain disease incidence with the operation
of activated sludge plants. Mr. Lee Hiser, L. L. Hiser and Associates,
New Braunfels, Texas, has also been consulted with regard to the conduct
of the survey. Mr. Riser's expertise is primarily in the area of design
and operation of sewage treatment facilities and his expertise was
valuable in determining a typical activated sludge sewage treatment
plant for use in the survey.
Consultation has also been made with the Metropolitan Sanitary
District of Greater Chicago with Mr. Bart Lyman, its general superintendent.
Additional consultations have been made with Mr. Raymond Rimkus,
Chief of Maintenance and Operations, and Dr. Lue-Hing, Director of
Research and Development of the Metropolitan Sanitary District of
Greater Chicago for their cooperation and assistance in the conduct
of the surveys near one of their activated sludge sewage treatment plants.
5. Estimation of Respondent Reporting Burden
The estimation of respondent reporting burden for the Current
Health Status Questionnaire is six minutes. This estimate is based on a
preliminary pre-test of randomly selected Southwest Research Institute
employees.
253
-------
6. Sensitive Questions
All of the questions on the Health Status Questionnaire, except
for the first question, maybe considered sensitive. Questions 2, 3,
and 4 ask about chronic illnesses and/or diagnosed diseases and certain
medications the subject may be taking with regard to their illnesses.
It is necessary to obtain information regarding chronic illnesses and/or
diagnosed diseases in order to provide an assessment of the subject's
state of health, both before the sewage treatment plant becomes
operational and after it is on-line. Additionally, it is necessary since
some trace metal surveys have shown that certain chronic diseases
significantly affect trace metal levels in different tissues. Medication
taken by individuals has been shown to affect trace metal levels, making
this type of correlation important. Question 5 relates to recent symptoms
experienced by the subject. This information is essential in order to
provide an assessment of the health effects associated with a sewage
treatment plant.
254
-------
APPENDIX B
SURVEY BACKUP MATERIALS
Recruitment handouts, interviewer instruction and information sheets, and the news release used to
generate public awareness of the program are presented in this Appendix.
255
-------
HOW ABOUT AN EXTRA $75 ?
WE NEED 250 PAID VOLUNTEERS FOR A RESEARCH STUDY
Sum-nary
As you may be aware, the Metropolitan Sanitary District of Greater Chicago is
constructing an advanced wastewater reclamation facility on Salt Creek, adjacent
to your community. Southwest Research Institute (SwRI) of San Antonio, Texas
is performing a study for the U. S. Environmental Protection Agency (EPA)
on the possible incidence of disease resulting from the operation of sewage
treatment plants. Although no unusual incidence of disease from the operation
of the Salt Creek Plant is anticipated, SwRI will be collecting air, soil, and
water samples within the study area shown on the map. Additionally, samples
of blood, urine, feces, sputum and hair clippings will be taken from paid
volunteers.
Participants Needed
250 participants, both male and female, are needed, age requirements being
3 months to any inter ested elderly.
Samples Needed from Each Volunteer Participant
Four sampling periods will span eighteen months necessitating:
(1) Four overnight urine samples
(2) Four overnight fece samples
(3) Four blood samples
(4) Four hair samples
(5) Four sputum samples
Sample Collection
Four sampling periods will span an eighteen-month period beginning, tentatively,
October 14, 1974. Volunteers will be chosen from a Household Health Survey
conducted September 23 through 27, 1974. Those volunteers fitting appropriate
age and locational categories will be notified by mail of their selection and where
to meet for orientation and provision of sample containers. Medical personnel
will collect the blood and sputum samples and a professional barber will collect
the hair samples.
Payment for Volunteer Participants
Each complete sampling period will earn each participant $12. 50. For those
completing all four sampling periods, a bonus of $25 will be paid. Participants
in all four sampling periods can earn $75. A family of four can earn $300.
WORTH $75 TO EACH PARTICIPANT
256
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INTERVIEWER HANDOUT-INNER SECTORS
Introduction and Summary
Hello, my name is_ and I represent Southwest Research
Institute which is under contract to the U. S. Environmental Protection Agency.
SwRI is conducting a study to assess the health effects resulting from the operation
of sewage treatment plants.
As you may be aware, the Metropolitan Sanitary District of Greater Chicago is
constructing an advanced wastewater reclamation facility on Salt Creek adjacent
to your community. Although no unusual incidence of disease from the operation
of the Salt Creek Plant is anticipated, SwRI will be collecting air, soil, and
water samples within your community to obtain baseline information. Additionally,
SwRI is conducting this Household Survey to obtain further information on the
Household incidence of disease before the Salt Creek Plant becomes operational.
After the plant is operational, a second survey will be performed with the results
of both being compared.
Individual Health Participant Survey
Your household is in an area of particular interest for our study. We would like
to know if you would be interested in being a paid participant in an individual
health survey. If you are interested, this information sheet explains what the
individual health survey is all about.
We can fill out this participant questionnaire now, or I can leave it with you for
later retrieval by me this week, or you can mail it to SwRI with the enclosed,
self addressed, stamped envelope no later than September 28, 1974.
All members of your household can participate, so I will leave you sufficient
participant questionnaires for your whole family.
257
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INTERVIEWER INFORMATION - PARTICIPANT SECTORS
SUMMARY
Southwest Research Institute of San Antonio, Texas, under contract to the
U. S. Environmental Protection Agency, and in cooperation with the Metropolitan
Sanitary District of Greater Chicago, will be conducting a study to assess the
health effects resulting from the operation of sewage treatment plants. Focus of
the study is a three-mile radius area surrounding the Salt Creek Water Reclamation
Plant presently under construction by the Metropolitan Sanitary District of Greater
Chicago. In this study, air, water, and soil samples will be gathered and analyzed
and the incidence of disease in people within the study area will be monitored to
establish a baseline. After the plant becomes operational, further sampling and
monitoring will take place for comparison.
YOUR PART
You, as an interviewer, are a key element in the success of this study.
Your cpntact with the members of the community in your assigned sector will
provide two very essential features. Your first and primary function is to obtain
public health information through the completed household questionnaire forms.
Each interviewer is expected to obtain at least fifty completed forms. Secondly,
during the course of each interview, a solicitation is made for attracting the
needed paid volunteers. Two hundred and fifty paid volunteers are needed for
a successful study, and you will receive a bonus for each volunteer you obtain
(completed Health Survey Participant Questionnaire forms).
INSTRUCTIONS
If the respondant replies in the negative to question 10, and in the positive
to question 15, on the Household Health Survey, you will provide him with a
copy(ies) of the Health Survey Participant Questionnaire which the participant(s)
will fill out immediately if time permits. If it is more convenient, arrange to
pick the completed form(s) up at a later time. In all cases, interested participants
will receive informational forms.
Note: The information listed under : "Interviewer Note and Record" must be
completed for each form. Distance and sector information will be indicated on the
sector maps provided to you. It is particularly importent to fill out this informa-
tion on the "Health Survey Participant Questionnaire" forms left with potential
participants. Completed forms need to be coded and turned in daily, at the end
of the day or at the beginning of the next day. This will allow proper logging and
inspection of the forms in an orderly manner by the survey team leader.
PAYMENT
Each interviewer will be paid for working a forty-hour week. At least
fifty (50) forms are expected from each interviewer, completed and properly coded.
For Health Survey Participant forms, a bonus of as much as $4. 00 per form will
be paid. $1000 has been set aside for 250 forms at $4. 00 each. If more than
250 volunteers are obtained, the $1000 will be prorated accordingly. (Example: A
total of 333 volunteers would place the bonus at $3. 00 per form). You will be
paid by Manpower, Inc. for the 40 hour week. You will be paid directly by SwRI
for any bonus you earn.
258
-------
INTERVIEWER INFORMATION - OUTLYING SECTORS
SUMMARY
Southwest Research Institute of San Antonio, Texas, under contract to the
U. S. Environmental Protection Agency, and in cooperation with the Metropolitan
Sanitary District of Greater Chicago, will be conducting a study to assess the
health effects resulting from the operation of sewage treatment plants. Focus of
the study is a three-mile radius area surrounding the Salt Creek Water Reclamation
Plant presently under construction by the Metropolitan Sanitary District of Greater
Chicago. In this study, air, water, and soil samples will be gathered and analyzed
and the incidence of disease in people within the study area will be monitored to
establish a baseline. After the plant becomes operational, further sampling and
monitoring will take place for comparison.
YOUR PART
You, as an interviewer, are a key element in the success of this study. Your
contact with the members of the community in your assigned sector will provide
public health information through the completed household questionnaire forms.
Each interviewer is expected to obtain at least fifty completed forms.
INSTRUCTIONS
If the respondant replies in the negative to question 10, and in the positive
to question 15, .on the Household Health Survey, you will provide him with a copy
of the informational forms. Encourage the individuals as much as possible to
volunteer; this information is very valuable to the success of the study.
Note: The information listed under: "Interviewer Note and Record" must
be completed for each form. Distance and sector information will be indicated
on the sector maps provided to you. Completed forms need to be coded and turned
in daily, at the end of the day or at the beginning of the next day. This will allow
proper logging and inspection of the forms in an orderly manner by the survey
team leader.
PAYMENT
Each interviewer will be paid for working a forty-hour week. At least
fifty(SO) forms are expected from each interviewer, completed and properly coded.
You will be paid by Manpower, Inc. for the 40 hour week.
259
-------
NEWS RELEASE
Southwest Research Institute of San Antonio, Texas, under contract
to the U. S. Environmental Protection Agency, and in cooperation.
with the Metropolitan Sanitary District of Greater Chicago, will be
conducting a study to assess the health effects resulting from the
operation of sewage treatment plants. The focus of the study is a
three-mile circular area (see map) surrounding the Salt Creek Water.
Reclamation Plant under construction by the Metropolitan Sanitary
District of Greater Chicago.
The project will obtain baseline information by gathering air, water,
and soil samples, and monitoring the incidence of disease in people
within the study area. After the plant becomes operational, further
sampling and monitoring will take place for comparison.
The study area encompasses portions of Schaumburg and Elk Grove
Townships in Cook County, and Bloomingdale and Addison Townships
in DuPage County. The areas of Schaumburg, Hoffman, Estates,
Rolling Meadows, Elk Grove Village, Itasca, and Roselle, which fall
within the study boundaries will be surveyed on a house-to-house basis
with the tentative survey date set as September 9-13, 1974. Based
upon the Household Health Survey, volunteers will be selected to
participate in an individual health survey. Individual volunteers will
be paid for participation in a sampling program which will span
approximately eighteen months, tentatively scheduled for commencement
on September 30, 1974. Of 1, 000 households surveyed, SwRI hopes
to obtain 250 volunteers.
Four camplings will be taken at various time intervals, with volunteers
receiving $12. 50 for each sampling period. A volunteer participating
in all four samplings can earn a total of fifty dollars.
260
-------
APPENDIX C
LETTERS OF ACCEPTANCE/NON-ACCEPTANCE TO SURVEY PARTICIPANTS
The letters to individuals advising them of whether they had been selected as participants are presented in
this Appendix.
261
-------
SOUTHWEST RESEARCH INSTITUTE
8500 CULEBRA ROAD • POST OFFICE DRAWER 28510 • SAN ANTONIO, TEXAS 7828-J
Division of Chemistry
and Chemical Engineering
August 22, 1975
Dear Survey Participant:
The questionnaire, which you recently completed for
Mrs. Carol McArthur, has been reviewed,and you have been selected
for our research program. This is an important environmental study
being conducted by Southwest Research Institute under contract to
the U. S. Environmental Protection Agency, and, without the help of
volunteers such as you, could not be conducted. Your participation
is very much appreciated.
The survey team will be collecting samples on September 4, 5,
and 6, 1975. To make the sampling as convenient for you as possible,
we are trying to make arrangements to collect these samples at the
Lexington Green. You will be notified on August 28 or 29 regarding
the exact times and location for sampling. The total time required
for your participation will be kept as short as possible (20 minutes
total);and you will receive your money at the time of sample collections.
Mrs. Carol McArthur (894-5577) will be contacting you for
further information. If you plan to be out of town on these dates,
please notify us.
Yours very truly,
i Donald E.'Johnson, Ph.D.
v Project Manager,
Environmental Survey Team
SAN ANTONIO, HOUSTON, CORPUS CHRISTI, TEXAS, AND WASHINGTON. D.C.
262
-------
SOUTHWEST RESEARCH INSTITUTE
8500 CULEBRA ROAD • POST OFFICE DRAWER 28510 • SAN ANTONIO, TEXAS 78280
Division of Chemistry and
Chemical Engineering
August ZZ, 1975
Dear Survey Participant:
The questionnaire which you recently completed for Mrs. Carol
McArthur has been reviewed by our staff. The requirements of this
important environmental survey conducted by Southwest Research
Institute under contract to the U.S. Environmental Protection Agency
are many. One such requirement for this study is the number of people
which can be selected in a particular age group. An over abundance of
questionnaires were completed for your particular age group. Therefore,
we could not include all of these in the study, and this is the reason tor
your not being selected.
Your questionnaire will be set aside for the present. However,
should any other participant in your age category withdraw from the
survey, you shall be contacted to fill this vacancy.
Your willingness to participate in the study is appreciated.
Very truly yours,
?<••>_ Donald E. Johnson
Project Manager
Environmental Survey Team
DEJ:pb
SAN ANTONIO. HOUSTON, CORPUS CHRIST!, TEXAS. AND WASHINGTON. D.C.
263
-------
APPENDIX D
VOLUNTEER'S INFORMED CONSENT FORM
The form executed by the volunteers indicating that they were informed of the nature of the study and
gave consent to participating in it is presented in this Appendix.
264
-------
SOUTHWEST RESEARCH INSTITUTE
8500 CULEBRA ROAD • POST OFFICE DRAWER 28510 • SAN ANTONIO. TEXAS 78284
VOLUNTEER'S INFORMED CONSENT
I,
residing at
hereby acknowledge and certify to the following:
1. That I hereby volunteer and consent to participate as a
human test subject in an experiment designed to determine exposure to
environmental pollutants entitled, "Health Implications of Sewage
Treatment Facilities".
Z. That I have been given, in my opinion, an adequate
explanation of the nature,duration and purpose of the experiment, the
means by which the experiment will be conducted and any possible in-
conveniences, hazards, discomforts, risks, and adverse effects on my
health which could result from my participation therein;
3. That I have been informed of all appropriate alternative
procedures, if any exist, that might be advantageous to me;
4. That I understand my questions concerning procedures
which affect me will be answered fully and promptly;
5. That I understand that I have the right to withdraw my
consent and to discontinue participation in this experiment at any
time without prejudice regardless of the status of the experiment and
regardless of the effect of such withdrawal on the objectives and
results which the experiment is designed to achieve; and I also under-
stand that my participation in the experiment may be terminated at
any time by the investigator in charge of the project or the physician
supervising the project regardless of my wishes to the matter;
6. That I hereby understand and agree that the samples
collected from me will be analyzed for their content of trace metals,
bacterial, and viral pathogens and that these are the only tests that
will be made on these samples and that no medicinal compounds
will be analyzed.
over
SAN ANTONIO, HOUSTON, CORPUS CHRIST), TEXAS. AND WASHINGTON, D.C.
265
-------
Volunteer's Informed Consent
Page 2
7 . That I attained the age of years on my last birthday
which was , and that I
am executing this Volunteer's Informed Consent as my free act
and deed.
Executed this th day of
19 .
Executed in my presence and in
the presence of each other .
Witness Volunteer
Person informing volunteer and
obtaining volunteer's consent
If subject is a minor, complete the following:
Subject is a minor - age .
Father Mother
Guardian Other person and relationship
266
-------
APPENDIX E
INSTRUCTION SHEET
The instruction sheet given to the participants advising them of proper collection techniques for the
clinical samples is presented in this Appendix.
267
-------
I
SOUTHWEST RESEARCH INSTITUTE
8500 CULEBRA ROAD • POST OFFICE DRAWER 28510 • SAN ANTONIO. TEXAS 78284
INSTRUCTION SHEET
The success of this research project depends on your complete
cooperation. Only through the assistance of volunteers, such as you,
can we obtain information on environmental pollutants. Your samples
will also provide a free check on your current health status. Incomplete
sampling of urine, feces and sputum will produce incomplete information,
so please follow these instructions.
Urine Collection
(1) You will be given a urine collection bottle that has been
cleaned in a special manner. Please avoid introducing anything else into
this bottle other than urine.
(2) Beginning in the evenings (approximately 5:00 P.M.), you
will begin collecting all of your urine in the container provided. Please
urinate into this container each time you urinate until approximately
8:00 A.M. the next morning.
(3) The collection bottle should be capped tightly after each use.
Feces Collection
(1) You will also be given a collection container for feces
that has been cleaned in a special manner. Your feces will be the only
contents placed in this container.
(2) The portable toilet bag assembly has been provided
for your convenience in feces collection. Open the sample bag and
place it on the portable seat as shown in the instructions included with
the kit. As with the urine collection, begin collecting all of your feces
at approximately 5:00 P.M. and continue until the following morning.
(3) When collecting your specimen, be sure not to mix any
urine or toilet paper with the feces. Analysis cannot be done on a
sample that has been mixed with urine or paper. Also, take great
care not to contaminate the outside of the collection vessel with feces
because there may be potentially serious consequences to those handling
the specimen. The specimen bag is to be sealed with the twist ties
and then placed in the polyethylene container designated for feces.
...over, please.
SAN ANTONIO. HOUSTON, CORPUS CHRISTI, TEXAS, AND WASHINGTON, D.C.
268
-------
Instruction Sheet
Page Z
(4) There is no restriction on your diet. You can eat or
drink anything you wish, and you can eat breakfast before coming to
work in the morning.
Sputum Collection
(1) You will be given a collection container that has been
cleaned in a special manner for your sputum. Remove the lid from this
container only when collecting your sputum and then immediately
replace the lid.
(Z) Upon arising in the morning, cough deeply at least
several times . The material collected on the back of the tongue or in
the mouth is to be placed in the container provided.
If you are unsuccessful in producing a specimen after
coughing deeply, lie across a bed with your head and chest slanting
downward toward the floor for 10 or 15 minutes. Coughing after this
procedure will very likely result in producing an acceptable specimen
for collection.
Current Health Status Questionnaire
Please fill out the questionnaire at home. This will save time at
the collection center.
Please report to the collection center at the time designated for
blood, hair and throat swab collections. Please bring your urine,
feces and sputum specimens and the completed current Health Status
Questionnaire with you.
Thank you very much for your help.
269
-------
APPENDIX F
CERTIFICATION OF APPRECIATION
A copy of the certificate given to each study participant is presented in this Appendix.
270
-------
Certificate of Appreciation
Witfi sincere tfianks, tfiis certificate of appreciation is fiere6y presented to
for participation in an Snvironmental Protection Agency survey of tfie
environment and tfie fiealtfi of persons living near tfie $onn 6. Sgan
Wasteivater Reclamation Plant. 9t is only ffirougfi tfie efforts of concerned
persons, sucfi as yourself, tfiat a study of tfiis nature can Se completed.
Sfiis certificate is presented as a token of our gratitude.
Presented at ScfiaumSurg, Illinois, tfiis 8tfi day of October, 1976.
Donald E. Johnson, PhD.
Project Leader
Department of Environmental Sciences
Soutfiiuest Research institute
-------
APPENDIX G
TYPICAL CODED DATA REPORTING FORMS
Table Title
A-1 AEROSOL RUN REPORT ON METEOROLOGY AND SAMPLING CONFIGURATION
A-2 UTSA-CART AEROSOL RUN ANALYSIS REPORT
A-3 HOUSEHOLD HEALTH SURVEY
A-4 HEALTH SURVEY PARTICIPANT QUESTIONNAIRE
A-5 CURRENT HEALTH STATUS QUESTIONNAIRE
A-6 METAL ANALYSIS CODING FORM
A-7 PATHOGEN ANALYSIS CODING FORM
A-8 PATHOGEN ANALYSIS DATA FORM
272
-------
TABLE A-l AEROSOL RUN REPORT ON METEOROLOGY AND SAMPLING CONFIGURATION
Project 01-4007 Chicago, Illinois
Aerosol Run Number
Run Date r
Field Supervisor
u
Time
Estimated wind angle &
Estimated pasquill stability class
Estimated cloud cover
Wind velocity from weather station mph
Temperature from weather station °F
Relative humidity from weather station %
Temperature of effluent
Beginning of
Sampling
GboO
E 44.17S"
End of
Sampling
0-1
5Z
-5Z
Sampler Distance
in Meters
Upwind
as:
.50 ;
LVAS Sampler
Number
Personnel Running
Samplers
Remarks:
(continued)
273
-------
TABLE A-l (continued)
SALT CREEK WATER RECLAMATION PLANT
Soil and Water Sampling Sites
Key
1. Digesters
2. Tliickcner Building
3. Maintenance Building
4. Pre treatment Building
5. Control Buildin"
6. Laboratory Building
7. Filter Building
SCALE
I cm ^ 6O METERS
8. Settling T?.nks No. 1
9. Aeration Basins No. •i
10. Aeration Basins No. 2
11. Settling Tanks No. 2
® Soil Sampling Site
O Water Sampling Site
(continued)
274
-------
TABLE A-l (continued)
; C
275
-------
TABLE A-2 UTSA-CART AEROSOL RUN ANALYSIS REPORT
Jlcpu
KupurLtrtl
Run Number
I-O
Run Date and Time
Wastewater Analysis:
SwRI Label Code.
Total Coliphage Count
Solids Fraction
Liquid Fraction
Aerosol Analysis:
Sampler
SwRI Label Code
Bacteria
Proteus
Pcuedomonas, cfu/lOOml
Streptococcus, cfu/lOOml
Salmonella, cfu/lOOml
Shigella, cfu/lOOml
Viruses
3 Day Plaque Count, pfu/ml
5 Day .Plaque Count, pfu/ml
Coliphase
Coliphage Count, pfu/ml
COMMENTS:
- 1-7
Kcports to: f)nv:1tt C.'ihi.inti, San Antonio
James HagIstor,
pfu/1
*#
S.Ox
\.SxiO
/- SOP fati}
o.o
-------
TABLE A-2 (continued)
...£. °*
Kurt No.
flerosol Analyses:
Sampler
SwRI Label Code
Bacteria
Proteus, cfu/lOOml
Psuedomonas, cfu/lOOml
Streptococcus, cfu/lOOml
Salmonella, cfu/lOOml
Shigclla, cfu/lOOml ^C5g^(i>lfc3 ^-Soo fab)
•Viruses
3 Day Plaque Count, pfu/ml ^
5 Day Plaque Count, pfu/ml ^O.MgJD L.O.
Coliphage
Coliphage Count, pfu/ml
COMMENTS:
NOTATION: TNTC-too numerous to count
ND-none detected
CS-contaminated sample
277
-------
TABLE A-3 HOUSEHOLD HEALTH SURVEY
Form Approved
OMB 158-R0117
STAFF USE ONLY
Cols 1-4
10 #
HOUSEHOLD HEALTH SURVEY
Expires July 1977
Cols 5-30
Cols 31-50
Cols 51 -65
Cols 66-70
Cols 71-77
Col 80
m
Col 5
3. How many persons reside in your household? .
For each person i
to the youngest:
your household, inctuding yourself, please indicate the age and tex beginning with the oldest and proceeding
Cols6-7 Cots 8-9 Cols 10-11 Cols 12-13 Cols 14-15 Cols 16-17 Cols 18-19 Cols 20-21 Cols 22-23 Cols6-23
Person , 2 3456789
Col 24 Col 25 Col 26 Col 27 ColjS Col 29 Col 30
5. Has any one of your household ever been diagnosed as having any of the following chronic illnesses:
Col 31 Cot 32 Cols 24-32
D D D
(Person in Household)
3 4 56 7
tuberculosis
malignancies
asthma—hay fever
diabetes '" '"
heart conditions
hypertension
chronic sinusitis & bronchitis
arthritis & rheumatism
rheumatic heart disease
thyroid ditease (specify type)
liver disease (specify type)
kidney disease (specify type)
6. During the past year, has anyone of your household been diagnosed as having any of the following diseases?
n Household)
5 6
polio
infectious jaundice
pneumonia
worms
skin disease
pleurisy
spinal meningitis
influenza
cioop
sleeping sickness
dysentery
empyema
Cols 40-49
Cols 50-59
Cols 60-69
Cob 70-79 Col 60
Cols 10-19 [5]
Cols 20-29
Cols 30-39
Cols 40-49
Cols 50-59
Cols 60-69
Cols 70-79 Col 80
Cols 10-19 [~3l
Coli 20-29
Cod 30-39
Coll 40-49
Cols 50-59
Col. 60-69
Coli 70-79 Col 80
Cols 10-19 [4]
Cols 20-29
Coli 30-39
Coll 4(M9
Coti 50-59
Co I, 60-69
Coll 70-79 coigQ
7. Has anyone in your household experienced any of the following symptoms during the past three months:
SI
(Person in Household)
34561
severe headache not relieved by aspir
severe dizziness
severe pain in bones and joints with
high fever
severe weight loss
severe night sweats
hemorrhagic rash
canker sores around mouth
yellow eyeballs
sore throat
cough
cold
Cols 10-19
Cols 20-29
Colt 30-39
Cols4O-49
Cols 50-59
Cols 70-79 Col 80
Cols 10-19 [6]
Cols 20-29
Cols 30-39
Cols 4049
(continued)
278
-------
TABLE A-3 (continued)
bodv
diarrhea
bloody diarrhea
bloody urine
nausea
vomiting
pain in chest on deep breathing
weakness o< arms or legs
cough up blood
stiff neck with lever
stiff neck with rash
general weakness
draining ear
fever above 103*F
severe trouble with teeth
colicky pains in abdomen
brown urine
shortness of breath
convulsion)
unconsciousness (not due to blow o
head)
yellow skin
How long have you:
Lived at your present address
Months
51 ?:11
"3 4
more than No
24-35 3ytys Response
"
Coll 50-59
Col! 60-69
Coll 70-79
Coll 10-19
Coll 20-29
Coll 30-39
Coll 40-49
Coll 50-59
Coll 60-69
Coll 70r79
Coll 10-19
Coll 20-29
Coll 30-39
Coll 4049
Coll 50-59
Coll 60-69
Coll 70-79
Coll 10-19
Coll 20-29
Coli 3039
Coll 40-49
Coll 50-59
Cols 60-69
Coll 70-79
Col 80
13
Col 80
©
Col 80
IS
Col 80
m
is have you and your family changed living quarters during the last five years?
1-No
Ji-Yes, window only
11. How many bedroi
n your home?
A--
12. What is the usual oc
Occupation
Professional
Administrative
Technical
Clerical (sec'y, ' 'le
clerk, cashier)
Sales
cupation of the head of your household?
Code
01
02
03
04
05
Occupation
F orema n/Craf tsman
Laborer (kitchen,
custodial, maintenance)
Student
Volunteer Worker
Waiter, waitress
Code
06
13. What educational level has been completed by the head of your household:
1-less than 8(h grade
2 -8th grade
3-high school-incomplete
4-high school-completed
5-college-incomplete
6-college-completed
7-graduate school
Occupation
Teacher
Librarian
Housewife
Public assistance
(policeman, fireman)
Retired
14. Distance from John Egan Treatment Plant (refer to map for coding A-K)
a. freeway, expressway or turnpike
b. other major multilane traffic artery
c. large industrial operation
17. IMNOWO 123
less than
2blocks
less than
1 mile
279
-------
TABLE A-4 HEALTH SURVEY PARTICIPANT QUESTIONNAIRE
Fonli A;»[iruvt!£) ROI IV
HEALTH SURVEY PARTICIPANT QUESTIONNAIRE
STA
ID#
FF USE ONLY
Cols 1-4
0
(p
s
a
of Birth_
/o
_Month_
I 9
4. What is your sex?
5. What is your marital status?
1-male
'TWie
•—'
2-married
3-separated
-Day .
£n
Year
4-divorced
5-widowed
Jflhich of these best describes your present occupational status:
/l^nployed fulltime {including self-employed) 5-student
^•employed part-time 6-play/nursery school
3-unemployed 7-pre-schoo)
4-housewife
7. What is your usual occupation? (please specify) _
8-retired A y^) /^ I
8. If you are employed, what is the nature of the company for which you work? (please specify) .
9.
10.
m
How many hours of the day do you normally spend more than 2 miles from your home?
What is the natural color of your hair? 1-brown (4^lond
2-black 5-gray
3-red
Have you ever smoked as many as five packs of cigarettes, thaxjj, as many as 100 cigarettes during your
entire life? 1-yes |2-r»
Do you now smoke cigarettes?
1-yes
1.1
15.
How many years have you lived in your present city or town? \& tylQ • vears
At your present address? \@ Ml^fV 'years
How many times have you changed living quarters during the last five years? _
Do you plan to move during the coming year? 1-yes f^"^
Have you ever been diagnosed as having any of the following chronic illnesses:
No
Presently receiving medication
Yes (please specify type)
Ast'ima-Hay Fever
(continued)
Cols b-30
Cols 31 50
Cols 51-65
CoIsG6-'/0
Cols 71-7 /
0 I i
LU
Col cl.'i
Col, Si l.i
Col 1 1
Cui \:-.
Cols M-lb
Cot» ic-i;
Cols 1K-10
Cois 2:1:'«
Cols 27-:'.-:
coi ro
CoK 30 :.i
280
-------
TABLE A-4 (continued)
Presently rficeivirvj modiciitiun
(please specify lype)
, •
\
1
H or to inn \
vr
1
\
\
17. Is your home air conditioned ? 1-no 2-yes, window only 3-yts, central
^•s /*\ ^*"1O t^ '
19. What educational level has been completed by the head of your household:
. i
,/-
,-.
,'
S'
1
1
x(
4
f'
f.
v"
0.
Jf
1
•\
Cois ID <\ i
Cot". f.'.' 44
Col-J 4't 4 /
Cols T: l,(j
Coii M •••.•;
Cols 'i--1.lii,
CoK b7-5'l
Coh 60 ii >
Coh C'l f,',
Col 60
Cols C7-Go
Col 69
2-8th grade 6-college completed
3-high school-incompleted /v-Jaduate school
4-high school-completed
YOU HAVE COMPLETED THE QUESTIONNAIRE
THANK YOU FOR YOUR COOPERATION
20. Distance '/ ': K">V c.. ,
21. Sector
22. Distance to nearest:
a. freeway, expressway or turnpike
b. other major multilane traffic artery
c. hrtjti industrial ope.'ation
23. IMNOWO 1 2 3 4\516
INTERVIEWER NOTE AND RECORD
less than 2 blocks less than 1 mile
one rrule or more
I \3\
Col 70
Col 7?
Coi '/':.
Col 7-t
• •
Col SO
281
-------
TABLE A-5 CURRENT HEALTH STATUS QUESTIONNAIRE
Form Approtwd
OMB No. 158-R0117
1. NvnadnfuU)
2. A»a you prnantly raMlvwiB
(continued)
282
-------
TABLE A-5 (continued)
5. Have you iiperienced any oi tht following lymptomt during: the p
Exwritf
(1 three month*' At promt?
ed during put E
wvere headache not relieved by Mprin
tevere diuinei*
lavare weight ton
cankar torn around mouth
l.3i;H ^13-14
I 0 I 0 I Coh 15-16
J^ I P I Coii 17-18
j~p~[ T^l Cot* 19-20
f/ 1 QJ Cwli 31-22
f 0 I 0 1 Colt 23-24
I | I J| I Coli25-2fi
Colt 29-X
Colt 31 32
Cod 33 34
Con 35-36
Coll 37-38
vomiting
ytllowikin
Dain in chMi on OMP brwlhing
wwknMt ol armi or )*p*
nilt r«ck wiih taw
draining wr
ftvw «bov« 103*F
MVtra troubU with tMth
colicky paint in abdomen
brown urine
ahonnetl ot breelh
corwurtiom
uneomekM*neci (not due to blow on heed)
Cob 61-62
Col* 63 64
Colt flS-66
Co H 67 -68
Colt 60-70
Col* 71-72
YOU HAVE COMPLETED THE QUESTIONNAIRE-THAN t( VOU FOR VOUR COOPERATION
283
-------
TABLE A-6 METAL ANALYSIS CODING FORM
1 HAIR
Cot O./T>
Pi a- S7
H* 0-17
METAL ANALYSIS CODING FORM
IS
i
?£
P
M
M
M
M
P
?
r-
M
F
M
F
F
M
F
Is
M
M
M
1?
Label Code
ID
umber
1 3
CO
0
0
0
0
0
0
/
/
/
/
/
/
/
/
/
. -0
•4
J
9
?
1^2
4
/
2
3
4
5
6
•7
*7
/
2
3
f
_f
£
/
9
i
9
(;
/
3
5
Sample medium |
5
rf
\
\
I
/
//
Sample analysis |
6
M
I
\
\
i
f
/
//
Site Code
7
/O
I
\
\
I
/
/
C
B <
//.
/
/
\
\
1
H.
Subper/Sample * I
10
5/
/
\
I
f /
Sub-lubperiod |
"
!!
4
1
'3
O
14
O
1
\
1
1
I
1
1
II
|
4-
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TABLE A-7 PATHOGEN ANALYSIS CODING FORM
ID j
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-------
TABLE A-8 PATHOGEN ANALYSIS DATA FORM
01-4007 PATHOGEN ANALYSIS P - POJ
N - NE(
Tissue: rec.e-«;
Analysis:
Participant
(W
4-^r/'
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(continued)
286
-------
TABLE A-8 (continued)
0 1 -4007 PATHOGEN ANALYSIS
P - POSITIVE
N - NEGATIVE
Tissue!
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Analysis:
•
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Specific
rrn-t?.U5
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Period #2 Period #4
Chance
ID 9
Oct. '74 Oct. '76
+/-
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117
131
141
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
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287
-------
APPENDIX H
INDIVIDUAL PARTICIPANT DATA TABLES
Table Title
A-9 BACTERIAL ISOLATIONS IN FECES SAMPLES FROM INDIVIDUAL PARTICIPANTS
A-10 BACTERIAL ISOLATIONS IN THROAT SWABS FROM INDIVIDUAL PARTICIPANTS
A-11 PARASITE ISOLATIONS IN FECES SAMPLES FROM INDIVIDUAL PARTICIPANTS
A-12 VIRAL ISOLATIONS IN FECES SAMPLES AND THROAT SWABS FROM INDIVIDUAL PARTICIPANTS
A-13 COXSACKIEVIRUS AND POLIOVIRUS ANTIBODY TITERS FROM ALL INDIVIDUAL PARTICIPANTS
A-14 ECHOVIRUS ANTIBODY TITERS BY HEMAGGLUTINATION INHIBITION FROM ALL INDIVIDUAL
PARTICIPANTS
A-15 ADENOVIRUS ANTIBODY TITERS BY SERUM NEUTRALIZATION FROM 100 INDIVIDUAL PARTICIPANTS
A-16 COXSACKIEVIRUS ANTIBODY TITERS FROM 100 INDIVIDUAL PARTICIPANTS
A-17 ECHOVIRUS ANTIBODY TITERS BY SERUM NEUTRALIZATION FROM 100 INDIVIDUAL PARTICIPANTS
A-18 ECHOVIRUS ANTIBODY TITERS BY HEMAGGLUTINATION INHIBITION FROM 100 INDIVIDUAL
PARTICIPANTS
A-19 REOVIRUS ANTIBODY TITERS BY HEMAGGLUTINATION INHIBITION FROM 100 INDIVIDUAL PARTICIPANTS
A-20 TRACE METAL CONCENTRATIONS IN BLOOD SAMPLES FROM INDIVIDUAL PARTICIPANTS (^g/100 ml)
A-21 TRACE METAL CONCENTRATIONS IN FECES SAMPLES FROM INDIVIDUAL PARTICIPANTS (jig/g)
A-22 TRACE METAL CONCENTRATIONS IN HAIR SAMPLES FROM INDIVIDUAL PARTICIPANTS (>ig/g)
A-23 TRACE METAL CONCENTRATIONS IN URINE SAMPLES FROM INDIVIDUAL PARTICIPANTS (/»g/l)
A-24 HEMATOCRIT VALUE OF BLOOD SAMPLES FROM INDIVIDUAL PARTICIPANTS
288
-------
SYMBOLS USED IN REPORTING INDIVIDUAL PARTICIPANT DATA:
P Positive
N Negative
NSC No sample was collected
IS Insufficient sample for analysis
NA Sample was not analyzed
U Unsatisfactory because nonspecific inhibitors were not removed by
kaolinizing and absorbing with human type "O" erythrocytes
289
-------
TABLE A-9 BACTERIAL ISOLATIONS IN FECES SAMPLES FROM INDIVIDUAL PARTICIPANTS
t-J
\O
O
Participant
I.D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
144
(continued)
Period
1
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
P
N
N
N
P
N
N
NSC
P
NSC
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
P
2
N
NSC
NSC
N
N
NSw
NSC
N
N
N
N
N
P
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
4
N
N
P
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
NSC
P
P
N
5
~N~
N
N
N
NSC
N
N
N
N
N
N
N
NSC
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
P
N
N
NSC
Pseudoranas
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
P
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
Period
2
N
NSC
NSC
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
P
N
N
P
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
1,
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
5
N
N
N
N
NSC
N
P
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
Perl£>d
1
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
TT
NSC
NSC
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
4
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
5
TT
N
N
N
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
Qiterobacter
Period
1
P
P
N
P
P
N
N
N
N
N
P
P
P
N '
N
P
P
N
P
N
N
N
NSC
N
NSC
P
P
P
P
P
P
P
P
P
P
N
P
P
P
N
N
N
P
N
P
N
N
2
P
NSC
NSC
P
N
NSC
NSC
N
N
N
N
P
N
P
N
P
N
P
N
N
NSC
N
P
P
N
NSC
N
NSC
N
P
N
N
N
N
N
N
N
N
N
P
NSC
NSC
P
N
N
P
P
4
P
P
P
P
P
P
P
NSC
N
P
P
N
P
N
P
N
P
P
N
P
P
N
N
P
N
P
P
P
P
P
N
P
P
P
N
N
N
NSC
NSC
NSC
NSC
NSC
N
NSC
P
P
N
5
P
N
P
P
ICC
N
N
N
N
N
P
N
-CC
P
N
N
N
N
P
P
P
N
N
P
P
P
P
P
N
P
P
N
P
P
P
N
P
NSC
NSC
NSC
NSC
NSC
P
N
N
P
NSC
-------
TABLE A-9 (continued)
to
VD
Participant
I.D.
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
241
(continued)
Proteus
Period
J^
N
N
N
N
N
N
N
N
N
N
P
P
N
N
P
N
N
N
N
N
NSC
N
P
P
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
2
N
N
P
N
N
NSC
P
N
N
P
P
P
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
P
N
P
N
N
N
P
N
N
N
N
N
P
P
N
N
4
N
N
N
N
NSC
NSC
N
N
N
NSC
N
N
P
NSC
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
P
NSC
NSC
N
N
N
N
N
N
N
N
5
N
N
N
N
N
P
N
N
N
NSC
P
N
N
P
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
P
P
N
N
N
N
N
P
N
N
N
Pseudononas
Salmonella
Period
1
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
2
N
N
N
N
N
NSC
N
P
N
NSC
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
P
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
5
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
2
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
5
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
Enterobacter
Period
1
P
P
N
P
P
P
N
P
N
P
P
P
P
P
N
P
N
N
P
P
NSC
P
N
N
N
P
P
P
P
P
P
P
N
N
P
P
N
N
P
P
N
P
P
P
NSC
N
P
_2_
N
P
P
N
P
NSC
N
P
N
P
P
P
P
N
N
P
N
N
P
P
N
N
P
N
P
P
P
N
NSC
N
N
P
N
N
N
N
P
P
P
N
N
P
N
N
N
N
N
4
N
P
N
P.
NSC
NSC
N
N
P
NSC
P
N
P
N
N
P
P
P
N
NSC
NSC
NSC
NSC
NSC . .
NSC
NSC
N
P
P
NSC
NSC
N
N
P
N
N
P
NSC
NSC
N
N
P
N
N
P
P
P
5
P
P
P
P
N
N
N
N
N
NSC
P
P
N
P
N
N
P
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
P
NSC
NSC
N
P
P
P
P
N
P
N
N
N
P
N
N
N
P
N
-------
TABLE A-9 (continued)
to
~-o
K)
Participant
I.D.
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
348
349
350
353
354
Period
1
N
N
N
N
N
N
N
N
N
P
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
2
P
N
NSC
P
N
N
N
N
N
N
N
N
N
N
N
P
N
P
N
N
N
NSC
N
N
N
N
N
N
P
N
P
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
4
P
N
N
NSC
N
N
N
NSC
N
N
N
N
N
NSC
N
N
NSC
P
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
5
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
P
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
Pseudcrronas
Period
1
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
P
N
N
P
2
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
P
NSC
N
N
N
N
N
N
N
N
N
P
4
TT
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
5
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
P
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
Salmonella
Biterobacter
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
4
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
5
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
P
N
N
N
NSC
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
NSC
N
Period
1
P
N
P
N
N
P
P
P
P
N
P
P
P
P
NSC
N
P
P
P
N
N
P
P
N
P
P
N
P
P
N
N
N
N
N
N
N
N
N
P
N
P
N
N
P
N
P
N
2
P
N
NSC
N
N
N
N
N
P
P
N
N
N
N
N
N
N
N.
N
N
N
NSC
P
P
P
N
P
P
P
N
P
N
N
N
N
P
NSC
N
N
N
N
P
N
N
N
N
N
4
N
N
P
NSC
P
P
N
NSC
P
N
N
N
N
P
P
N
N
P
P
N
NSC
NSC
P
P
N
P
N
P
P
P
N
P
P
N
N
P
N
N
N
NSC
NSC
N
P
N
N
N
P
5
P
NSC
N
NSC
NSC
NSC
NSC
NSC
P
N
P
P
N
P
N
N
N
P
P
P
NSC
N
N
P
N
N
NSC
N
P
P
N
N
N
N
P
N
N
P
N
P
P
P
P
N
N
NSC
N
(continued)
-------
TABLE A-9 (continued)
Participant
I.D.
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
413
414
415
416
417
418
(continued)
Proteus
Period
1
N
N
N
P
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
P
N
N
N
N
N
NSC
N
N
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
P
P
P
P
NSC
N
N
N
N
NSC
N
NSC
N
N
P
N
N
N
NSC
N
N
NSC
N
N
N
N
P
P
P
P
N
4
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
NSC
NSC
N
N
N
N
N
5
N
N
N
P
N
NSC
N
N
N
N
N
P
P
N
P
N
N
NSC
NSC
N
N
NSC
P
N
N
N
N
NSC
NSC
NSC
NSC
N
N
P
NSC
N
N
N
N
N
NSC
NSC
N
NSC
N
NSC
NSC
Pseudccnonas
Period
1
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
2
TT
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
P
N
NSC
NSC
N
NSC
N
N
N
P
NSC
N
NSC
N
N
N
N
N
N
NSC
N
N
NSC
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
5
~N~
N
N
N
N
NSC
N
N
N
N
N
N
P
N
N
N
N
NSC
NSC
N
N
NSC
N
N
P
N
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
P
P
N
NSC
NSC
N
NSC
N
NSC
NSC
Salmonella
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
NSC
N
NSC
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
NSC
N
N
NSC
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
'N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
5
N
N
N
N
N
NSC
N
N
P
N
N
N
N
N
N
N
N
NSC
NSC
N
N
NSC
N
N
N
N
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
NSC
N
NSC
NSC
Enterobacter
Period
1
N
P
P
N
P
N
P
P
P
N
N
P
P
P -
P
P
P
P
P
P
P
P
NSC
NSC
P
P
P
N
N
P
P
P
P
P
P
N
P
P
P
P
P
NSC
P
P
N
P
N
2
~N~
N
P
P
N
N
P
P
N
' N
N
P
N
NSC
N
N
N
N
N
N
NSC
N
N
P
N
NSC
N
NSC
N
P
P
N
P
P
NSC
P
P
NSC
N
P
N
N
P
P
P
P
N
4
P
N
P
P
N
NSC
P
N
N
P
N
N
N
P
N
P
NSC
NSC
NSC
N
N
P
N
P
N
N
N
N
P
P
P
N
N
P
NSC
P
P
P
NSC
P
NSC
NSC
N
N
N
N
N
5
N
N
P
N
N
NSC
N
N
P
N
N
N
N
N
P
N
N
NSC
NSC
P
N
NSC
P
N
P
P
N
NSC
NSC
NSC
NSC
N
P
N
NSC
N
N
N
N
N
NSC
NSC
N
NSC
N
NSC
NSC
-------
TABLE A-9 (continued)
Participant
I.D.
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
468
500
501
502
503
504
505
506
507
508
509
510
512
(continued)
Proteus
1
P
P
P
NSC
N
P
N
N
N
P
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
HSC
NSC
NSC
Per:
2
N
P
N
N
P
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
N
N
NSC
NSC
P
N
N
N
N
LOd
4
N
N
N
.N
N
N
N
N
N
NSC
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
17
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
KSC
N
N
N
N
N
NSC
5.
P
N
N
P
N
N
N
N
N
NSC
N
N
P
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
NSC
N
N
NSC
NSC
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
Pseudcmonas
Enterobacter
Period
1
P
P
N
NSC
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
N
NSC
NSC
NSC
NSC
NSC
2
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
SSC
NSC
N
NSC
P
N
NSC
NSC
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
5
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
NSC
N
N
NSC
NSC
N
NSC
NSC
NSC
NSC
N'
N
N
NSC
N
. N
N
N
1
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
N
NSC
NSC
NSC
NSC
NSC
Period
2
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
NSC
N
N
NSC
NSC
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N.
N
N
N .
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
5
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
NSC
N
H
NSC
NSC
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
Period
1
N
N
N
NSC
P
N
P
N
N
P
P
P
N
N
P
P
P
P
P
N
N
N
P
N
N
N
P
P
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
NSC
NSC
NSC
2
N
P
P
N
N
P
NSC
P
P
N
P
N
NSC
NSC
N
N
P
P
N
N
N
N
N
N
N
N
N
P
N
P
P
P
NSC
N
NSC
N
N
NSC
NSC
N
N
P
P
P
4
-r
P
P
P
N
N
N
P
N
NSC
N
P
P
N
N
N
N
N
1
N
N
N
P
P
N
P
N
N
N
NSC
NSC
N
NSC
NSC
NSC
NSC
P
N
N
N
N
P
N
N
5
N
N
P
P
N
N
N
N
N
NSC
P
P
P
P
NSC
P
NSC
NSC
NSC
NSC
NSC
P
N
N
P
N
NSC
N
N
NSC
NSC
N
NSC
NSC
(EC
NSC
P
N
N
NSC
N
P
N
N
-------
TABLE A-9 (continued)
to
^o
Ln
•Participant
I.D.
600
601
602
603
604
605
607
608
609
610
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
628
629
630
631
632
633
634
637
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
Proteus
Period
3
N
N
N
N
N
N
P
N
N
NSC
N
N
P
N
N
N
N
N
N
N
N
N
N
P
N
P
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
P
N
N
N
N
N
N
NSC
IBC
5
TT
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
NSC
N
N
N
NSC
NSC
NSC
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
P
P
NSC
N
N
N
N
N
Pseudonunas
3
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
P
N
N
N
N
N
N
NSC
NSC
Period
5
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
NSC
N
N
N
NSC
NSC
NSC
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
Salmonella
Period
3
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N"
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
5
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
N
N
N
N
N
NSC
N
N
N
NSC
NSC
NSC
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
P
N
N
N
Enterobacter
Period
3
N
N
P
P
N
N
P
P
P
NSC
P
P
P
P
P
P
N
N
P
P
P
N
N
NSC
P
P
N
N
P
N
N
N
N
P
NSC
P
N
P
P
N
P
P
P
N
N
P
P
NSC
NSC
5
N
P
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
NSC
P
P
N
NSC
NSC
NSC
N
N
N
N
N
NSC
N
NSC
N
N
P
N
N
N
N
N
N
NSC
N
N
P
P
N
-------
TABLE A-10 BACTERIAL ISOLATIONS IN THROAT SWABS FROM INDIVIDUAL PARTICIPANTS
Participant
I.D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
144
(continued)
Streptoaoccus-alpha
Period
1
T ~
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
2
~N~
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N .
N
P
N
N
N
N
N
N
N
N
N
N
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N •
N
N
N
N
N
N
N
N
P
N
N
P
N
NSC
NSC
NSC
NSC
' NSC
P
N
P
P
P
5
N
P
P
N
P
N
N
N
N
N
.N
•N
N
N
N
P
N
P
P
N
N
N
N
N
N
N
N
P
.N
N
N
P
P
P
P
P
P
NSC
NSC
NSC
NSC
NSC
P
P
P
P
P
Streptococcus-beta
Period
1
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
NSC
P
NSC
P
P
P
P
P
P
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
- TT— —
NSC
NSC
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
—
N
P
N
N
N
P
N
P
P
P
N
P
P
P
P
P
P
P
P
P
N
P
N
N
N
P
P
P
P
P
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
5
T5
P
P
p
p
p
p
p
p
P
p
p
P
p
p
p
p
P
p
p
P
P
p
p
p
p
p
N
P
P
P
P
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
Streptccoocus-ganma
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
use
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
P
N
P
N
P
2
. N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
P
N
P
N
N
N
P
N
P
N
P
N
P
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
P
P
P
P
N
NSC
NSC
NSC
^
NSC
N
N
N
P
P
5
N
N
N
N
N
N
N
N
N
P
N
N
N
N
P
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
P
P
P
N
NSC
NSC
NSC
NSC
NSC
N
P
P
P
N
Staphylococcus aureus
Period
1
N
N
P
N
N
N
N
P
P
N
P
N
N
N
N
N
P
P
P
P
N
N
NSC
N
NSC
P
N
P
N
P
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
P
NSC
NSC
N
N
N
N
N
N
P
P
N
P
P
N
N
P
P
P
P
N
P
P
P
N
N
N
N
P
N
P
P
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
4
P
N
N
N
N
N
N
P
P
P
N
P
N
N
N
N
P
P
P
P
P
P
N
N
N
N
P
P
P
P
P
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
P
N
N
N
P
5
P
N
N
P
N
P
P
P
P
N
N
P
P
P
N
N
P
N
N
P
P
P
P
N
P
P
P
N
P
P
P
N
P
N
N
N
P
NSC
NSC
NSC
NSC
NSC
P
N
N
N
P
-------
TABLE A-10 (continued)
Participant
I.D.
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
241
(continued)
Streptooo
ccus-alphe
1
Period
1
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
NSC
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
NSC
P
P
2
P
P
P
P
P
P
P
P
P
NSC
N
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
P
P
N
P
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
4
P
P
P
P
P
N
N
N
N
NSC
P
P
P
P
P
P
P
N
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
P
P
NSC
NSC
P
P
P
P
N
N
P
P
P
P
N
P
P
P
P
P
5
P
P
P
P
P
P
N
P
P
NSC
P
P
P
P
P
P
P
P
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
P
P
NSC
NSC
P
P
P
P
P
P
P
P
P
P
N
N
P
P
P
P
Streptoooccus-beta^
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
Period
2
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
5
P
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
P
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
Streptococcus-gang
Period
1
N
P
P
P
N
N
P
N
P
N
P
N
N
N
N
P
N
N
P
P
NSC
N
P
N
P
P
P
P
N
N
N
N
N
N
P
P
P
P
N
N
P
N
N
N
NSC
P
N
2
N
N
N
P
P
P
P
N
N
NSC
N
P
P
P
P
P
P
N
N
P
N
P
N
N
N
N
P
N
N
P
P
N
N
P
N
P
P
P
N
N
P'
P
P
N
P
P
P
4
N
N
N
P
P
P
P
N
N
NSC
P
P
P
N
N
P
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
P
P
NSC
NSC
P
N
P
N
N
N
P
P
N
P
N
N
P
N
N
P
5
N
N
P
P
P
N
N
P
P
NSC
P
P
N
P
P
N
P
P
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
P
P
NSC
NSC
N
P
P
N
P
P
N
P
P
P
N
N
N
P
P
P
Staphylpcoocus aureus
Period
1
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
'N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
2
N
N
N
N
N
N
P
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
P
N
N
N
N
N
P
N
N
P
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
P '
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
P
N
P
N
5
N
P
N
N
N
P
N
N
NSC
N
N .
P
N
N
P
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
P
N
N
P
N
N
N
P
N
P
N
N
N
-------
TABLE A-10 (continued)
Participant
I.D.
242
243
244
254
255
256
257
258
260
262
263
266
267
269
273
275
276
278
284
290
293
294
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343 .
348
349
350
353
354
Streptococcus-alpha
Period
1
p
N
P
P
P
P
P
P
P
P
N
P
P
P
NSC
P
P
P
N
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
2
P
P
P
NSC
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P,
P
P
P
P
P
P
P
P
P
P
4
P
N
P
NSC
P
P
P
P
P
N
P
N
P
N
P
P
P
N
P
N
NSC
N
P
N
P
P
P
P
N
P
P
P
P
N
N
N
N
P
N
N
NSC
N
N
P
P
P
N
5
P
N
P
NSC
NSC
NSC
NSC
NSC
P
P
P
P
P
P
P
P
P
N
P
P
NSC
P
P
P
P
P
P
N
P
P
P
P
N
P
N
N
P
P
P
P
P
P
P
P
P
P
N
Strep toooocus-beta
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
N
P
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N .
N
N
N
N
N
4
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
P
N
N
N
N
5
N
P
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
jStxejatoooccus-gamna
1
P
N
N
N
P
N
P
P
P
N
P
P
P
N
NSC
P
N
N
P
P
N
P
N
P
P
N
N
P
P
N
N
P
N
N
P
P
P
P
N
N
P
N
P
N
N
N
N
2
P
P
P
NSC
P
P
P
P
P
P
P
N
P
P
P
P
N
P
N
P
P
P
N
P
P
P
P
N
N
P
P
P
P
P
P
P
N
P
P
P
P
N
P
P
P
P
P
Period
4
P
N
P
NSC
P
N
P
P
P
P
N
P
N
P
P
N
P
N
N
N
NSC
P
N
P
P
P
P
P
N
P
P
P
P
N
N
N
N
P
P
N
NSC
N
P
P
P
N
P
5
P
N
P
NSC
NSC
NSC
NSC
NSC
P
P
N
N
P
P
P
P
P
N
P
P
NSC
P
P
P
P
N
P
N
P
P
P
P
N
P
N
N
P
N
P
N
N
P
P
P
N
P
N
Stuttjiyloooccus aureus
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
2
N
P
P
NSC
N
N
P
N
N
N
P
N
N
N
N
N
P
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
5
N
P
N
NSC
NSC
NSC
NSC
NSC
N
N
P
P
N
N
N
N
N
P
N
N
NSC
N
N
N
N
P
N
P
N
N
N
N
N
N
P
P
N
N
N
N
N
N
N
N
P
N
P
(continued)
-------
TABLE A-10 (continued)
Participant
I.D.
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385 '
386
387
388
390
391
392
393
394
395
396
397
398 .
399
400
402
403
405
406
407
408
412
413
414
415
416
417
418
(continued)
Streptococcus-alpha
Period
1
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
NSC
P
P
P
P
P
P
P
NSC
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
2
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
P
NSC
NSC
NSC
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
N
P
P
P
P
P
P
4
P
P
N
P
P
P
P
P
P
P
P
P
P
P
P
P
NSC
NSC
NSC
P
P
N
N
N
N
P
P
N
N
P
N
N
P
P
NSC
N
P
N
P
P
NSC
NSC
P
P
• N
P
P
5
N
P
P
P
P
N
P
P
P
P
P
P
P
P
P
P
P
NSC
NSC
P
P
P
N
P
P
P
P
NSC
NSC
NSC
NSC
N
P
P
NSC
P
P
P
P
P
NSC
NSC
P
P
P
P
P
S treptpooocus-beta
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
"N
N
N
N
N
N
N
N
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
P
P
P
5
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
P
NSC
NSC
P
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
N
NSC
NSC
P
N
N
P
N
Streptococcus-gamna
Period
1
N
P
N
P
P
N
P
N
N
N
P
N
N
P
P
P
N
N
P
N
P
N
P
NSC
N
N
N
N
P
P
P
NSC
P
N
P
N
N
N
P
P
N
N
P
N
P
N
P
2
P
P
N
P
P
P
P
N
P
N
P
N
N
P
P
N
NSC
NSC
NSC
P
N
N
P
P
N
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
N
4
— '
P
N
N
N
N
P
P
N
P
P
P
P
P
P
P
NSC
NSC
NSC
P
P
N
N
N
N
P
N
N
N
N
P
N
P
P
NSC
N
N
N
P
P
NSC
NSC
P
P
N
P
P
5
N
P
P
P
P
N
P
P
P
P
P
N
P
P
N
P
N
NSC
NSC
N
P
P
N
P
P
P
N
NSC
NSC
NSC
NSC
N
P
P
NSC
P
P
P
P
N
NSC
NSC
N
P
P
N
P
Staphylocopcu% ,aureu_s_
Period
1
N
N
N
N
N
P
N
P
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
-------
TABLE A-10 (continued)
Participant
I.D.
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454 .
455
457
458
459
460
461
464
405
466
467
500
501
502
503
. 504
505
506
507
508
509
510
512
Streptococcus-alpha
Period
1
P
p
P
NSC
P
P
P
P
p
P
P
P
p
P
P
P
P
P
P
P
P
p
P
p
P
P
P
P
P
p
P
NSC
P
P
P
P
P
P
P
P
P
P
f
2
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
P
P
P
N
P
P
P
NSC
P
P
P
P
NSC
N
P
P
P
P
P
4
P
N
N
P
P
P
P
P
P
NSC
P
P
P
P
P
P
P
P
P
P
P
N
P
P
N
P
P
P
P
NSC
NSC
P
NSC
NSC -
NSC
P
P
N
N
N
P
P
P
5
P
P
P
P
P
N
N
P
P
NSC
P
P
P
P
P
P
P
P
NSC
NSC
NSC
P
P
P
N
P
P
N
P
NSC
NSC
P
NSC
NSC
NSC
P
N
P
P
N
N
P
P
Streptococcus-beta
Period
1
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
NSC
N
N
N
P
P
N
N
P
N
N
N
N
N
N
P
N
N
N
N
NSC
NSC
N
NSC
NSC
NSC
N
N
N
N
P
N
N
N
5
N
N
N.
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
NSC
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
P
N
jtreptoooccus-gamna
Period
1
p
N
P
NSC
P
P
N
N
N
N
N
P
P
N
P
N
tl
N
N
P
N
N
P
N
P
P
N
P
P
N
P
NSC
N
N
N
P
N
N
N
N
P
N
N
2
N
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
N
N
P
P
P
P
N
N
NSC
P
N
N
P
NSC
N
P
N
P
P
P
4
N
N
N
N
N
N
N
N
P
NSC
N
P
N
P
P
N
P
P
P
P
P
P
N
N
P
N
P
P
P
NSC
NSC
P
NSC
NSC
NSC
N
P
N
N
P
P
P
P
5
P
P
P
P
P
N
N
P
N
NSC
P
P
N
N
P
P
P
N
NSC
NSC
NSC
P
P
P
N
N
P
N
P
NSC
NSC
N
NSC
NSC
NSC
N
N
P
P
N
N
N
P
Staphyloooccus aureus
1
N
N
N
NSC
N
N
N
N
N
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
P
N
N
N
Period
2
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
P
P
N
N
N
N
N
N
N
N
P
N
N
N
N
NSC
N
N
N
N
NSC
P
P
P
N
N
N
4
N
N
N
N
N
N
N
N
P
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
5
N
N
N
N
N
P
N
N
P
NSC
N
N
P
N
N
N
N
P
NSC
NSC
NSC
N
N
N
P
P
N
P
N
NSC
NSC
N
NSC
NSC
NSC
P
P
N
N
P
N
N
N
(continued)
-------
TABLE A-10 (continued)
Participant
I.D.
600
601
602
603
604
605
606
607
608
609
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
628
629
630
631
632
633
634
637
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
655
Streptococcus-alpha
Period
3
P
p
N
'N
p
P
p
p
P
N
N
N
N
N
P
P
P
P
P
P
N
P
P
P
P
P
P
P
P
N
P
P
NSC
P
P
P
N
P
P
NSC
P
P
P
P
P
P
P
N
N
P
5
P
P
P
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
P
P
P
NSC
N
NSC
P
P
P
NSC
NSC
NSC
P
P
P
p
P
P
P
NSC
P
P
P
P
P
P
P
P
P
P
P
. NSC
P
P
P
NSC
Streptoaoccus-beta
Period
3
N
N
N
N
N '
N
P
N
N
N
N
N
N
N
N
N
N-
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
NSC
N
N
N
N
N
N
NSC
N
N
P
N
N
N
N
P
N
N
5
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
N
NSC
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
P
N
N
P
N
N
NSC
N
N
N
NSC
Streptococcus-ganna
Period
3
P
P
N
N
N
N
P
N
N
N
N
P
P
N
P
P
P
N
P
N
N
N
N
N
N
N
N
P
N
P
N
N
NSC
N
N
N
N
P
N
NSC
N
N
N
P
N
P
N
P
P
P
5
P
P
P
P
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
P
P
P
NSC
N
NSC
P
P
P
NSC
NSC
NSC
P
P
P
N
P
N
P
NSC
P
P
N
N
P
N
N
P
N
P
P
NSC
P
P
P
NSC
Staphyloooccus
Period
3
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
P
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
aureus
5
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
P
N
N
N
NSC
N
NSC
N
N
N
NSC
NSC
NSC
N
N
N
P
N
P
N
NSC
N
N
N
P
N
N
P
N
N
N
N
NSC
N
N
N
NSC
-------
TABLE A-l 1 PARASITE ISOLATIONS IN FECES SAMPLES FROM INDIVIDUAL PARTICIPANTS
Participant
I.D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
29
30
31
32
33
45
49
50
74
75
76
77
78
79
80
82
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
(continued)
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
N
NSC
NSC
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
4
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
NSC
N
NSC
NSC
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
NSC
N
N
5
N
NSC
N
NSC
NSC
N
N
N
N
NSC
N
NSC
NSC
..N
NSC
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
302
-------
TABLE A-ll (continued)
Participant
I.D.
134"
145
146
157
158
159
160
162
165
166
171
172
177
178
179
180
181
182
183
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
239
240
241
242
243
244
252
254
255
256
257
(continued)
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
p5
N
N
N
NSC
N
NSC
N
N
2
N
NSC
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
NSC
N
NSC
N
N
N
4
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
NSC
NSC
P6
N
N
N
NSC
NSC
NSC
N
N
5
NSC
N
N
N
N
N
N
NSC
N
N
N
NSC
N
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
p5
N
NSC
N
NSC
NSC
NSC
NSC
NSC
303
-------
TABLE A-ll (continued)
Participant
I.D.
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
348
349
350
•353
354
355
356
359
361
362
363
364
365
366
367
(continued)
1
N
NSC
N
N
N
NSC
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
Period
2
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
N
N
N
N
N
N
P2
N
N
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
4
N
N
NSC
N
N
N
NSC
N
N
N
N
N
N
NSC
NSC
N
N
NSC
N
N
N
P2
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
5
NSC
N
NSC
N
N
N
N
NSC
NSC
NSC
NSC
N
N
NSC
N
N
N
N
N
NSC
N
p2
N
N
N
N
N
N
P5
N
N
NSC
N
NSC
N
N
N
N
NSC
N
• N
• N
Pi
N
NSC
NSC
N
N
.N
N
304
-------
TABLE A-ll (continued)
Participant
I.D.
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
413
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
(continued)
1
N
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
p3
N
N
N
N
N
N
N
N
N
NSC
N
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
Period
2
N
N
N
NSC
N
N
NSC
NSC
NSC
N
NSC
N
N
N
N
N
N
NSC
N
N
N
N
N
N
NSC
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
4
N
N
N
P2
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
NSC
N
N
N
5
N
NSC
N
NSC
NSC
NSC
N
NSC
NSC
N
N
NSC
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
NSC
N
N
N
N
N
NSC
NSC
N
NSC
N
NSC
NSC
N
N
N
NSC
N
N
NSC
N
N
NSC
N
N
N
305
-------
TABLE A-ll (continued)
Participant
I.D.
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
512
Period
1
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
NSC
N
NSC
NSC
N
N
NSC
N
N
N
NSC
2
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
NSC
N
N
N
N
N
4
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
P"
5
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
NSC
N
N
NSC
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
NSC
N
N
N
N
Period
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615
(continued)
3
N
N
N
N
N
N
NSC
N
N
N
NSC
N
N
N
N
N
5
N
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
NSC
NSC
NSC
N
N
306
-------
TABLE A-l 1 (continued)
Participant
I.D.
616
617
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
635
636
637
638
639
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
Period
3
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
NSC
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
5
NSC
N
N
N
NSC
N
N
N
NSC
NSC
NSC
NSC
N
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
iChilonHstax mesnili
2Entamoeba coli
3Entamkoeba hartananni
''Endolimax nana
5Giardia-lambia
6Trichononas
307
-------
TABLE A-l 2 VIRAL ISOLATIONS IN FECES SAMPLES AND THROAT SWABS
FROM INDIVIDUAL PARTICIPANTS
Participant
I.D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
(continued)
Feces (Pioornaviruses)
Period
1
~W
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
—
NSC
NSC
P
' P
N
NSC
N
N
N
N
N
N
N
N
N
N.
N
N
N
N
N
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
4
-P~
P
P
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
5
IT
p
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
NSC
N
N
NSC
NSC
Throat Swab (All viruses)
Period
1
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
•N
N
N
N
N
2
-FT
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
• N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N .
N
4 .
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
• N
N
N
NSC
N
N
N
N
N
N
N
• N
N
NSC
NSC
5
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
• N
N
N
N
N
NSC
• NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
NSC
308
-------
TABLEA-12 (continued)
Participant
I.D.
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
241
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
(continued)
Feces (Pioornaviruses)
Period
1
N
N
M
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
NSC
N
N
2
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
P
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
NSC
N
N
N
4
NSC
NSC
NSC
NSC
NSC
' N
N
N
NSC
NSC
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
5
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
P
N
P
N
N
N
N
N
N
N
NSC
N
NSC
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
N
NSC
N
P
N
NSC
N
N
NSC
N
N
NSC
N
N
N
N
P
N
N
N
N
N
N
N
N
N
N
Throat Swab (All viruses)
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
4
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
5
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
- N
N
N
N
N
N
N
.N
NSC
NSC
NSC
NSC
NSC
N
NSC
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
309
-------
TABLE A-12 (continued)
Participant
I.D.
348
349
350
353
354
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
363
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
413
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
(continued)
Feces (Pioomaviruses)
Period
1
N "
N
N
N
N
N
N
N
N
N
N
N
N
NSC.
N
N
NSC
N
N
NSC
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N .
N
N
N
N
N
N
N
N
N
NSC
N •
NSC
N
N
N
NSC
N •
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
N
N
2
N
N
N '
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
NSC
NSC
N
NSC
N
N
N
N
N
N
NSC
N
N
N
N
N .
N
NSC
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N •
N
NSC
NSC
N
N
4
N
N
N
N
• N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
P
N
NSC
NSC
N
N
N
N
N
P
P
N
N
N
N
N
N
P
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
5
N
N
. N
NSC
NSC
N
N
N
N
N
NSC
N
N
NSC
N
N
N
N
N
N
NSC
N
NSC
NSC
N
N
NSC
N
N
N
NSC
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
NSC
N
NSC
NSC
NSC
P
N
N
P
N
NSC
N
N
NSC
N
N
N
N
NSC
N
Throat Suab (All viruses)
Period
1
N
N
N
N
N
N
N
N
N
N
N
N ,
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
N
N
N
N
N
N
N
N
N
2
N .
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N .
N ' '
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N '
N
N
NSC
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
NSC
5
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
.N
N
N
NSC
NSC
N
N
N
N
N
N
N
N
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
N.
. NSC
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
•N
310
-------
TABLEA-12 (continued)
Participant
I.P..
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
512
Feces (Picomaviruses)
Period
m
N
N
N
N
N
N
N
N
.N
N
N
N
NSC
N
N
NSC
N
NSC
NSC
N
N
NSC
NSC
N
N
N
2
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
NSC
N
N
N
N
N
4
TT
N
N
N
N
' N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
sic
NSC
NSC
NSC
NSC
N
N
N
N
N
NSC
N
N
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
NSC
N
N
N
N
Throat Swab (All viruses)
Period
1
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
2
IT
N
N
N
N"
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
NSC
N
N
N
N
N
N
4
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
5
N
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
N
NSC
N
NSC
NSC
NSC
N
N
N
N
N
N
N
N
Period
600
601
602
603
604
605
606
607
608
609
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
637
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
3
N
N
N
N
NSC
N
NSC
N
N.
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
NSC
NSC
5
NSC
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
N
NSC
N
N
N
NSC
NSC
NSC
NSC
N
N .
N
N
N
NSC
N
NSC
N
N
N
N
N
N
N
N
N
NSC
N
N
N
P
N
TEeriod
3
TT
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
NSC
N
5
TT
N
N
N
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
N
N
N
N
NSC
N
NSC
N
N
N
NSC
NSC
NSC
NSC
N
N
N
N
N
N
N
NSC
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
311
-------
TABLE A-l 3 COXSACKIEVIRUS AND POLIOVIRUS ANTIBODY TITERS FROM ALL INDIVIDUAL PARTICIPANTS
Participant
I.D.
1
2
3
4
5
6
9
10
•11
12
13
14
15
16
18
19
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
144
Coxsackievirus B-l*
Poliovirus Type It
10
NSC
NSC
10
10
40
10
10
20
10
NSC
10
40
5
NSC
10
10
20
10
10
10
10
NSC
NSC
NSC
NSC
NSC
10
NSC
NSC
10
NSC
NSC
NSC
NSC
NSC
NSC
Period
1
10
100
100
100
100
100
10
100
100 •
10
10
10
10
10
100
10
10
10
100
100
100
100
100
100
100
100
100
NSC
100
100
100
NSC
100
10
10
100
100
10
100
100
10
100
100
10
10
100
100
2
10
NSC
NSC
100
100
100
10
100
100
10
10
10
100
<10
100
10
10
10
100
100
100
100
100
.100
100
100
100
NSC
100
100
100
100
100
10
10
100
100
10
100
100
10
100
100
100
10
100
100
4
10
100
100
100
100
100
NSC
100
100
10
10
10
100
10
100
10
10
10
100
100
100
100
100
100
100
100
100
NSC
100
100
100
100
100
10
10
100
100
NSC
NSC
NSC
NSC
NSC
100
100
10
100
100
5
NSC
100
100
100
100
100
10
100
100 •
10
10
100
100
10
100
10
10
10
100
100
100
100
100
100
100
100
NSC
NSC
100
100
100
100
100
10
10
100.
100
NSC
NSC
NSC
NSC
NSC
NSC
100
10
100
100
PolicR/irus Type
Period
1
100
'100
100
• 10
100
100
100
100
100
10
10
10
10
100
100
10
10
100
100
100
100
100
100
100
10
100
10
NSC
100
100
100
NSC
100
10
10
10
100
10
100
100
100
100
100
100
10
100
100
2
"100
NSC
NSC
10
100
100
100
100
100
10
100
10
100
100
100
10
10
100
100
100
100
100
100
100
10
100
10
NSC
100
100
100
100
100
10
10
10
100
10
100
100
100
100
100
100
10
100
100
4
100
100
10
10
100
10
NSC
100
100
10
100
10
100
100
100
10
10
100
100
100 .
100
100
100
100
100
100
10
NSC
100
100
100
100
100
10
10
10
100
NSC
NSC
NSC
NSC
NSC
100
100
10
100
100
• 5
NSC
10
10
10
100
100
100
100
10
10
100
10
100
100
100
10
10
100
100
. 100
100
100
100
100
10
100
NSC
NSC
100
100
100
100
100
10
10
10
100
NSC
NSC
NSC
NSC
NSC
NSC
100
10
100
inn
Poliovirus
TVpe3t
Period
1
100
10
10
10
10
10
10
10
100
<10
10
100
10
100'
100
10
10
100
100
100
100
100
100
10
100
100
NSC
100
100
100
NSC
10
10
10
100
100
10
100
100
100
100
100
100
10
100
100
2
100
NSC
NSC
10
10
10
10
10
100
10
10
100
10
100
100
10
10
100
100
100
100
100
100
10
100
100
NSC
100
100
100
100
10
10
10
100
100
10
100
100
100
100
100
100
10
100
100
4 .
100
10 .
10
10
10
10
NSC
10
100
<10
10
100
10
100
100
10
10
100
100
100
100
100
100
10
100
100
NSC
100
100
100
100
10
10
10
100
100
NSC
NSC
NSC
NSC
NSC
100
100
1C
100
100
5
NSC
10
10
<10
10
10
10
10
100
• 10
100
100
10
100
100
10
10
100
100
100
100
100
100
10
100
NSC
NSC
100
. 100
100
100
10
10
10
100
100
NSC
NSC
NSC
NSC
NSC
NSC
100
10
100
.100
(continued) .
-------
TABLEA-13 (continued)
Participant
I.D.
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
241
Coxsackievirus B-l*
PolioviruB Type
-go
NSC
10
2
To
10
NSC
20
NSC
10
NSC
NSC
NSC
10
10
10
20
20'
NSC
40
4
.40
10
NSC
40
10
-fo
NSC
40
10
10
:10
40
40
=10
10
:10
20
:10
=10
:10
=10
:10
NSC
NSC
NSC
NSC
NSC
NSC
NSC
10
<10
-------
TABLEA-13 (continued)
u>
£
Participant
I.D.
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
348
349
350
353
354
Coxsackievirus B-l*
2~
20
10
NSC
20
10
Period
~4
20
NSC
NSC
<20
10
10
5
20
NSC
NSC
NSC
NSC
NSC
10
10
10
NSC
20
NSC
10
NSC
10
10
NSC
NSC
NSC
NSC
NSC
NSC NSC <10
<10 NSC NSC
NSC <10 <10
10
10
Poliovirus Type 1"*"
Period
1
100
100
100
100
100
100
NSC
10
10
100
100
100
100
100
100
100
100
100
100
100
100
100
NSC
10
100
100
NSC
100
100
100
100
100
100
100
10
100
10
NSC
NSC
NSC
100
100
100
10
100
100
2
100
100
100
NSC
100
100
10
10
10
100
100
100
100
100
100
100
100
100
100
100
100
100
10
10
100
100
100
100
100
100
100
100
100
100
10
100
10
NSC
100-
100
100
NSC
100
10
100
100
4
loo
100
100
NSC
100
100
10
10
10
100
100
100
100
100
100
100
100
100
100
NSC
100
100
10
10
100
100
100
100
100
100
100
100
100
100
10
100
10
NSC
100
NSC
100
100
100
10
100
100
5
100
100
100
NSC
NSC
NSC
NSC
NSC
10
100
100
100
100
100
100
100
100
100
100
NSC
100
100
10
10
100
NSC
100
100
100
100
100
100
100
100
10
100
10
10
100
NSC
100
100
100
10
100
100
t
Polioviirus TVps 2
Period
1
Too
100
100
100
100
100
NSC
10
10
100
100
100
10
100
100
100
100
100
10
10
100
100
100
NSC
100
100
100
NSC
100
100
100
100
100
100
100
<10
100
10
NSC
NSC
NSC
100
100
100
10
10
10
2
100
100
100
NSC
100
100
100
10
10
100
100
100
10
100
100
100
100
100
10
10
100
100
100
10
100
100
100
10
100
100
100
100
100
100
100
10
100
10
NSC
100
100
100
NSC
100
10
10
10
4
100
100
100
NSC
100
100
100
10
10
100
100
100
10
100
100
100
100
100
10
10
NSC
100
100
10
100
100
100
10
100
100
100
100
100
100
100
10
100
10
NSC
100
NSC
100
100
100
10
10
10
5
100
100
100
NSC
NSC
NSC
NSC
NSC
10
100
100
100
10
100
100
100
100
100
10
10
NSC
100
100
10
100
100
NSC
10
100
100
100
100
100
100
100
10
100
10
10
100
NSC
100
100
100
10
10
10
Poliovirus
Type 3f
Period
1
100
100
100
100
100
100
NSC
10
10
10
100
100
100
100
100
100
100
100
100
100
100
100
NSC
10
10
100
NSC
100
100'
100
100
100
100
100
10
100
10
NSC
NSC
NSC
100
100
100
10
10
10
2
100
100
100
NSC
100
100
100
10
10
10
100
100
100
100
100
100
100
100
100
100
100
100
10
10
10
100
100
100
IOC
100
100
100
100
100
10
10
100
NSC
100
100
100
NSC
100
10
10
10
4
100
100
100
NSC
100
100
100
10
10
10
100
100
100
100
100
100
100
100
100
NSC
100
100
10
10
10
100
100
100
100
100
100
100
100
100
10
10
100
NSC
100
NSC
100
100
100
10
10
10
5
100
100
100
NSC
NSC
NSC
NSC
NSC
10
10
100
100
100
100
100
100
100
100
100
NSC
100
100
10
10
10
NSC
100
100
100
100
100
100
100
100
10
100
10
10'
100
NSC
100
100
100
10
10
10
(continued)
-------
TABLEA-13 (continued)
Participant
I.D.
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
413
414
415
416
417
418.
10
NSC
20
<10
•<10
NSC
NSC
Coxsackievirus B-l*
Period
2 4
10 tiSC
NSC <10
Polioyirus Type 1
t
20
NSC
NSC
NSC
NSC
NSC
10
NSC
NSC
NSC
NSC
NSC
NSC
10
10
10
NSC
NSC
10
10
NSC
NSC
10
NSC
NSC
NSC
NSC
10
NSC
NSC
10
Period
1
loo
100
100
10
<10
NSC
100
100
100
100
100
100
10
NSC
10
NSC
100
100
100
100
100
10
100
100
10
100
100
100
10
100
100
100
100
100
100
100
100
10
100
100
100
NSC
100
100
100
10
2
100
100
100
10
<10
NSC
100
100
100
100
100
100
10
NSC
10
NSC
NSC
NSC
NSC
100
100
10
100
100
10
100
100
100
10
100
100
100
100
100
100
100
100
<10
100
100
100
NSC
100
100
100
10
4
loo
100
NSC
10
<10
100
100
100
100
100
100
100
10
100
10
100
NSC
NSC
NSC
NSC
100
10
100
100
10
100
100
100
10
100
100
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
5
100
100
100
10
NSC
100
100
100
100
100
100
100
10
100
10
100
100
NSC
NSC
100
100
10
100
100
10
100
100
NSC
NSC
NSC
NSC
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
Poliovirus Type 2f
Period
1
150"
100
100
100
100
NSC
100
100
100
100
100
..10
100
NSC
10
NSC
100
100
100
100
100
100
100
100
10
100
100
10
10
100
100
100
100
100
100
100
100
10
100
100
100
NSC
100
100
100
10
2
100"
100
100
100
100
NSC
100
100
100
100
100
10
100
NSC
10
NSC
NSC
NSC
NSC
100
100
100
100
100
10
100
100
10
10
100
100
100
100
100
100
100
100
10
100
100
100
NSC
100
100
100
10
4
156"
100
NSC
100
100
100
100
100
100
100
100
.10
100
10
10
100
NSC
NSC
NSC
100
100
100
100
100
10
100
100
10
10
100
100
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
5
1(50"
100
100
100
NSC
100
100
100
100
100
100
10
100
10
10
100
100
NSC
NSC
100
100
100
100
100
10
100
100
NSC
NSC
NSC
NSC
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
Poliovirus Type 3*
Period
1
TOTI
100
100
100
100
NSC
10
100
100
100
100
100
10
NSC
10 •
NSC
100
100
100
100
10
10
100
100
10
100
100
10
10
100
100
100
100
100
100
100
100
10
100
100
100
NSC
100
100
100
10
2
IDO"
100
100
100
100
NSC
10
100
100
100
100
100
10
NSC
10
NSC
NSC
NSC
NSC
100
10
10
100
100
10
100
100
10
10
100
100
100
100
100
100
100
100
10
100
100
100
NSC
100
100
100
10
4
TOO"
100
NSC
100
100
100
10
100
100
100
100
100
10
100
10
100
NSC
NSC
NSC
NSC
10
10
100
100
10
100
100
10
10
100
100
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
5
TOT
100
100
100
NSC
100
10
100
100
100
100
100
10
100
10
100
100
NSC
NSC
100
10
10
100
100
10
100
100
NSC
NSC
NSC
NSC
100
100
100
NSC
100
100
10
100
NSC
NSC
100
100
100
100
10
(continued)
-------
TABLEA-13 (continued)
U)
OS
Participant
I.D.
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
512
Coxsackievirus B-l
Period
1 2 4 5
NSC
NSC
Poliovirus Type
<10 NSC <10
10 10 10
20
NSC
NSC
NSC
10
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
NSC
10
NSC
NSC
NSC
NSC'
NSC
Period
1
To
100
100
100
10
10
10
100
NSC
10
100
100
100
100
100
100
100
100
100
100
100
100
100
<10
10
100
100
100
100
100
100
100
100
100
100
100
NSC
100
100
100
10
100
NSC
2
in
100
100
100
10
10
10
100
100
10
100
100
100
100
100
100
NSC
100
100
10
100
100
100
<10
10
100
100
100
100
100
100
NSC
NSC
100
100
100
NSC
100
100
100
10'
100
100 '
4
10
100
100
100
10
10
10
100
100
NSC
100
100
100
100
100
100
100
100
100
100
100
100
100
10
10
100
NSC
100
100
NSC
NSC
NSC
NSC
NSC
NSC
NSC
100
100
100
100
10
100
100
5
To
100
100
100
10
10
10
100
NSC
NSC
100
100
100
100
100
100
100
100
NSC
NSC
NSC
100
100
10
100
100
100
100
100
NSC
NSC
NSC
NSC
NSC
NSC
NSC
100
100
100
100
10'
100
100
Poliovirus Type 2T
Period
1
10
10
100
100
10
10
10
10
NSC
10
100
100
100
100
100
100
100
100
10
100
100
100
10
10
100
100
100
100
100
100
100
10
100
100
100
100
NSC
100
100
100
10
10
NSC
2
10
10
100
100
10
10
10
10
100
10
100
100
100
100
100
100
NSC
100
10
100
100
100
01
10
100
100
100
100
100
100
100
NSC
NSC
100
100
100
NSC
100
100
100
10
10
100 .
4
10
10
100
100
10
10
10
10
100
NSC
100
100
100
100
100
100
100
100
10
100
100
100
10
10
100
100
100
100
100
NSC
NSC
10
NSC
NSC
NSC
100
100
100
100
100
10
10
100
5
10
10
100
100
10
10
10
10
NSC
NSC
100
100
100
100
100
100
100
100
NSC
NSC
NSC
100
10
10
100
100
100
100
100
NSC
NSC
10
NSC
NSC
NSC
100
100
100
100
100
10
10
100
Poliovirus Type 3^
Period
1
10
100
100
100
10
10
10
10
NSC
10
100
100
100
100
100
100
100
10
10
100
100
100
10
10
10
100
100
100
100
100
100
10
100
100
100
100
NSC
100
100
100
10
10
NSC
2
To
100
100
100
10
10
10
10
100
10
100
100
100
100
100
100
NSC
10
10
100
100
100
10
10
10
100
100
100
100
100
100
NSC
NSC
100
100
100
NSC
100
100
100
10
10
100
4
To
100
100
100
10
10
10
10
100
NSC
100
100
100
100
100
100
100
10
10
100
100
100
10
10
10
100
NSC
100
100
NSC
NSC
10
NSC
NSC
NSC
100
100
100
100
100
10
10
100
5
To
100
100
100
10
10
10
10
NSC
NSC
100
100
100
100
100
100
100
10
NSC
NSC
NSC
100
10
10
10
100
100
100
100
NSC
NSC
10
NSC
NSC
NSC
100
100
100
100
100
10
10
100
(continued)
-------
TABLE A-13 (continued)
Participant
I.D.
600
601
602
603
614
615
616
617
619
621
622
623
628
629
630
631
632
633
634
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
Coxsackievirus B-l*
Period
3 5
<10 20
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 *-10
<10 • <10
NSC <10
NSC <10
<10 <10
NSC <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
20 20
10 10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
<10 <10
NSC NSC
NSC NSC
NSC NSC
<10 <10
<10 <10
<10 <10
Poliovirus Type lf
Period
3
155
100
100
100
100
10
10
10
100
NSC
NSC
10
NSC
100
10
100
100
100
100
100
10
100
10
10
100
100
10
100
100
100
10
100
10
5
T65
100
100
100
100
10
10
10
100
10
10
10
10
100
10
100
100
100
10
100
100
100
10
10
100
100
10
100
100
100
10
10
10
Poliovirus Type
Period
3
TOO
100
100
10
100
10
10
10
10
NSC
NSC
100
NSC
100
10
100
100
100
100
10
100
10
100
10
100
100
100
100
100
100
100
100
2*
5
155
100
100
10
100
10
10
10
10
NSC
NSC
100
NSC
100
10
100
100
100
100
10
100
10
100
10
100
100
10
100
100
100
100
100
Poliovirus Type
Period
3
100
100
100
100
100
10
10
10
100
NSC
NSC
100
NSC
100
10
100
100
100
100
100
100
100
10
10
100
100
100
10
100
100
100
100
100
3+
5
155
100
100
100
100
10
10
10
100
100
100
100
100
100
10
100
100
100
100
100
100
100
10
10
100
100
100
10
100
100
100
100
100
*by Hanagglutinatlon Inhibition
tby Serum Neutralization
-------
TABLE A-14 ECHOVIRUS ANTIBODY TITERS BY HEMAGGLUTINAT1ON INHIBITION
FROM ALL INDIVIDUAL PARTICIPANTS
Echovirus 3
Participant
I. D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
Period
1
80
<10
10
10
<10
<10
20
<10
<10
U
160
80
40
10
<10
<10
<10
< I Q
<10
< 1 0
<10
<10
10
80
<10
<10
10
NSC
160
10
10
NSC
10
<10
<10
10 .
10
10
<10
<10
<10
<10
2
<10
NSC
NSC
<10
<10
<10
< 1 0
<10
< 1 0
80
<10
80
160
20
<10
10
<10
<10
<10
<10
<10
<10
20
20
<10
<10
<10
NSC
>320
160
<10
<10
10
<10
<10
<10
<10
20
<10
<10
<10
<10
4
20
<10
U
<10
<10
<10
NSC
<10
<10
U
160
40
160
10
10
<10
< 1 0
<10
< 1 0
<10
<10
<10
20
10
<10
<10
<10
NSC
>320
10
<10
<10
10
<1 0
<10
<10
<10
NSC
NSC
NSC
NSC
NSC
5
NSC
<10
<10
<10
<10
<10
<10
<10
20
U
160
20
160
<10
< 10
<10
<10
<10
< 10
<10
<10
<10
40
<10
<10
<10
NSC
NSC
40
10
. 20
<10
10
10
<10
<10
<10
NSC
NSC
NSC
NSC
NSC
1
10
<10
10
<10
<10
< 10
10
<10
40
U
80
<10
20
<10
10
< 1 0
<10
< 1 0
<10
< 1 0
<10
<10
10
<10
20
10
20
NSC
160
<10
10
NSC
<10
< 10
<10
<10
< 1 0
<10
^10
<10
<10
<10
Echovirus 7
Period
2
<10
NSC
NSC
<10
<10
< 10
<10
<10
20
10
10
<10
20
<10
<10
10
<10
<10
<10
<10
<10
<10
<10
<10
20
10
10
NSC
80
<10
<10
<10
<10
< 1 0
< 1 0
<10
< 1 0
10
<10
<10
<10
<10
4
10
<10
U
< 10
<10
< 10
NSC
<10
20
U
10
<10
20
<10
40
10
< 10
<10
<10
<10
< 10
<10
<10
<10
10
<10
10
NSC
40
< 10
<10
<10
<10 •
10
<10
<10
<10
NSC
NSC
NSC
NSC
NSC
5
NSC
< 10
< 10
< 10
<10
< 10
< 10
< 10
40
U
20
*10
20
<10
10
10
< 10
<10
<10
<10
<10
<10
<10
<10
20
< 10
NSC
NSC
40
< 10
<10
< 10
<10
< 10
<10
<10
<10
NSC
NSC
NSC
NSC
NSC
Echovirus 1 1
Period
1 2
<10 <10
<10 NSC
<10 NSC
< 10 < 10
10 < 10
10 < 10
10 < 10
10 < 10
10 < 10
U 10
80 40
< 10 < 10
80 80
<10 10
< 10 < 10
< 10 < 10
< 10 < 10
< 10 <10
< 10 < 10
<10 < 10
<10 < 10
<10 < 10
<10 10
<10 <10
<10 < 10
<10 < 10
20 <10
NSC NSC
160 80
<10 < 10
<10 < 10
NSC . <10
10 • 10
< 10 < 10
< | Q ^10
< 1 0 ^10
< 1 Q ^10
< 1 0 < 1 0
<10 <10
< 1 0 ^10
< 1 0 ^10
<10 <10
4
<10
< 10
U
< 10
< 10
< 10
NSC
< 10
< 10
U
40
< 10
160
< 10
10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
20
< 10
20 .
NSC
40
< 10
«10
< 10
10
< 10
< 1 0
< 10
< 1 0
NSC
NSC
NSC
NSC
NSC
5
NSC
< 1 0
< 10
< 1 0
< 10
< 10
< 10
< 10
< 10
U
160
< 10
160
< 10
< 10
< 10
< 10
< 10
< 10
< 10
<10
< 10
20
<10
< 10
<10
NSC
NSC
20
< 10
< 10
< 10
10
< 10
<10
< 10
<10
NSC
NSC
NSC
NSC
NSC
1
<10
< 10
< 10
< 10
< 10
< 10
20
< 10
40
U
40
< 10
< 10 •
< 10
< 10
< 10
< 10.
< 10
< 10
< 10
< 10
< 10
20
20
< 10
< 10
<10
NSC
160
< 10
< 10
NSC
< 10
< 10
10
10
10
10
10
10
10
10
Echovi
rus 12
Period
2
< 10
NSC
NSC
< 10
< 10
< 10
10
< 10
10
10
10
< 10
10
10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
10
< 10
10
< 10
< 10
< 10
NSC
160
< 10
< 10
< 10
10
< 10
< 10
< 10
<10
10
<10
<10
< 10
< 10
4
10
< 10
U
< 10
< 10
< 10
NSC
< 10
20
U
10
< 10
10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
10
< 10
< 10
< 10
NSC
40
< 10
< 10
< 10
10
< 10
< 10
< 10
< 10
NSC
NSC
NSC
NSC
NSC
5
NSC
10
10
10
10
10
10
< 10
20
U
10
< 10
20
< 10
< 10
< 10
< 10
< 10
< 10
< 10
< 10
10
10
< 10
< 10
10.
NSC
NSC
40
10
10
10
10
10
10
< 10
<10
NSC
NSC
NSC
NSC
NSC
(continued)
-------
TABLE A-14 (continued)
Participant
I.D.
131
141
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
Cchovirus 3
Period
1 2 4
<10 10 <10
20 20 20
10 <10 <10
<10 < 10 < 1 0
<10 < 10 <10
<10 <10 <10
<10 <10 <10
<10 < 1 0 < 1 0
<10 < 1 0 < 1 0
<10 <10 10
NSC <10 <10
10 20 20
<10 <10 <10
<10 <10 <10
<10 NSC NSC
NSC <10 <10
<10 10 10
<10 20 10
10 <10 <10
<10 <10 <10
<10 <10 <10
<10 <10 <10
<10 <10' <10
NSC <10 <10
40 20 NSC
10 20 NSC
<10 <10 NSC
<10 <10 NSC
<10 <10 NSC
<10 <10 NSC
NSC <10 NSC
20 10 10
<10 <10 <10
10 10 <10
<10 <10 NSC
NSC <10 NSC
80 40 20
10 <10 <10
<10 ^10 ^10
< 1 0 < 1 0 ^10
<10 ^10 ^10
<10 10 10
<10 <10 <10
5
NSC
40
10
< 10
< 10
< 10
<10
< 10
<10
<10
10
40
10
10
NSC
<10
<10
10
<10
<10
<10
<10
<10
<10
NSC
NSC
NSC
NSC
NSC
NSC
NSC
10
NSC
<10
NSC
NSC
40
<10
<10
-------
TABLE A-14 (continued)
1
<10
<10
40
<10
Echovirus 3
Period
2 4
<10 <10
10 10
10 10
<10 NSC
5
<10
10
10
NSC
Echovirus 7
Period
1245
<10 <10 <10 <10
<10 <10 <10 <10
<10 <10 <10 <10
<10 <10 NSC NSC
Cchovirus 11
Period
1245
<10 <10 10 <10
<10 <10 <10 <10
<10 <10 <10 <10
<10 10 NSC NSC
Echovirus 12
Period
1245
< 1 0 < 1 0 < 1 0 ^10
<10 ^10 ^10 ^10
< 1 0 < 1 0 ^10 ^10
<10 <10 < 10 <10
Participant
I.D.
227
228
229
230 '
232 NSC NSC <10 <10 NSC NSC <10 <10 NSC NSC <10 <10 <10 <10 <10 <10
235 NSC NSC NSC 20 NSC NSC NSC 10 NSC NSC NSC <10 <10 <10 <10 <10
242 10 20 20 20 <10 <10 <10 <10 <10 <10 10 10 <10 10 <10 10
243 < 10 <10 < 10 < 10 <10 <10 < 10 <10 <10 <10 <10 <10 < 10 <10 < 10 ~ < 10
244 < 10 < 10 < 10 < 10 <10 <10 < 10 <10 <10 <10 <10 <10 <10 <10 <10 < 10
254 <10 NSC NSC NSC <10 NSC NSC NSC <10 NSC NSC NSC <10 <10 NSC NSC
255 <10 <10 <10 NSC <10 <10 <10 NSC <10 <10 <10 NSC <10 <10
-------
TABLE A-14 (continued)
Echovirus 3 Echovirus 7 Echovirus 1 1 Echovirus 12
Participant Period Period Period Period
I.D. I 2 4 5 1 . 2 4 5 1245 124
338 <10 <10 <10 10 <10 <10 <10 <10 <10 <10 <10 <10 <10 <10 < I 0 <10
339 <10 <10 <10 <10 <10 <10 < 10 <10 <10 <10 < 10 <10 <10
-------
TABLE A-14 (continued)
Echovirus j , Echovirus 7 Echovirus 11 ~^^ EC ho virus. 12
Participant Period __ Period Period Period
I.D. 1 2 4 5 1 2 4 5 I 2 4 5 1 2 4
396 10 <10 <10 NSC <10 ^10 <10 NSC *10 <10 <10 NSC <10 *! 0 <10 ^10
397
-------
TABLE A-14 (continued)
Echovirus 3
Echovirus 7
Echovirus 1 1
Echovirus 1Z
Participant
I.D.
459
460
461
464
465
466
467
500
501
50Z
503
504
505
506
507
508
509
510
51Z
Period
1 Z 4
80 80 80
<10 <10 <10
<10 <10 NSC
<10 <10 <10
<10 < 1 0 *• 10
10 10 NSC
<10 <10 NSC
<10 NSC <10
<10 NSC NSC
<10 <10 NSC
<10 <10 NSC
<10 <10 <10
NSC NSC 10
<10 <10 40
<10 <10 10
<10 <10 <10
<10 <10 <1 0
<10 <10 <10
NSC <10 <10
5
160
<10
<10
<10
<10
NSC
NSC
<10
NSC
NSC
NSC
<10
ZO
40
<10
<1 0
<10
<1 0
10
Period
1 Z 4
<10 10 10
<10 <10 <10
<10 <10 NSC
<10 <10 <10
<10 <10 <10
<10 <10 NSC
<10 <10 NSC
<10 NSC <10
<10 NSC NSC
<10 <10 NSC
<10 <10 NSC
<10 <10 <10
NSC NSC <10
<1 0 ^10
-------
TABLE A-15 ADENOVIRUS ANTIBODY TITERS BY SERUM NEUTRALIZATION FROM 100 INDIVIDUAL PARTICIPANTS
Participant
I.D.
2
3
4
11
12
13
14
15
16
29
33
49
50
74
75
76
83
84
97
101
102
141
' 143
145
146
157
158
159
162
165
166
172
177
178
181
182
196
199
206
207
213
214
226
227
228
229
231
233
241
242
244
260
284
Adenovirus 1
Adenovirus 2
Adenovirus 3
Adenovirus 4
Adenovirus 5
•ao
10
10
10
10
10
10
10
10
10 10
10 10
10
10
10
10
10
10 10
<10 <10
<10 <10
<10 10
(continued)
-------
TABLEA-15 (continued)
Participant
I.D.
290
294
312
324
325
372
374
386
391
441
443
44S
446
457
464
465
500
504
510
Adenovirus 1
Period
Meroyirus 2
Period
Adenoyirug 3
Period
Adejiovirus 4
10
5
10
10
Period
Aienovirus 5
5
no
Period
5
no
10
10
10
Period
603
614
615
616
617
619
621
628
630
631
632
633
634
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
3
10
10
10
10
10
10
Period
3 5
TO no
10
Period
10
10
10
Period
3 5
no no
10
Period
10
10
10
10
10
10
-------
TABLE A-16 COXSACKIEVIRUS ANTIBODY TITERS FROM 100 INDIVIDUAL PARTICIPANTS
Participant
I.D.
2
3
4
11
12
13
14
15
16
29
33
49
50
74
75
76
83
84
97
101
102
141
143
145
146
157
158
159
162
165
166
172
177 '
178
181
182
196
199
206
207
213
214
226
227
228
229
231
233
241
242
244
260
284
290
Coxsackievirus A-7t
Period
1 5
<]_0
-------
TABLEA-16 (continued)
Participant
I.D.
~Z94~
312
324
325
372
374
386
391
441
443
445
446
457
464
465
500
504
510
603
614
615
616
617
619
621
628
630
631
632
633
634
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
Ooxsackievirus A-?*
Period
1 5
10
10
10
10
10
10
10
Period
10
<10
10
10
10
10
20
20
10
10
20
Coxsackievirus A-9t
Period
10 10
10 10
10 10
Period
3 5
<10
-------
TABLE A-l 7 ECHOVIRUS ANTIBODY TITERS BY SERUM NEUTRALIZATION
FROM 100 INDIVIDUAL PARTICIPANTS
Participant Ecftovirus 4 Echovirus 8 Echovirus 33
I.D. PeriodPeriodPeriod
1 5 15 1 5
2 <10 <10 <10 <10 <10
11
-------
TABLEA-17 (continued)
Participant Echovirus 4 Bchovicus 8 Echovirus 33
I.D. Period Period Period
1 " 5 J^_ 5 1 5
284 ^TO ^10 <10 710 <10 <10
290 <10 <10 <10 <10 <10 <10
294 <10 <10 <10 <10 <10 <10
312 <10 <10 <10 <10 <10 <10
324 <10 <10 <10 <10 <10 <10
325 <10 <10 <10 <10 <10 <10
372 <10 20 <10 <10 <10 <10
374 <10 <10 <10 <10 <10 <10
386 <10 <10 <10 <10 <10 <10
391 <10 10 <10 <10 <10 <10
441 <10 <10 <10 <10 <10 <10
443 <10 <10 <10 <10 <10 <10
445 20 10 <10 <10 10 <10
446 <10 <10 <10 <10 <10 <10
457 <10 <10 <10 <10 <10 <10
464 <10 <10 <10 <10 <10 <10
465 10 10 <10 <10 10 <10
500 <10 <10 <10 <10 <10 <10
504 <10 <10 <10 <10 <10 <10
510 <10 <10 <10 <10 <10 <10
Period Period Period
_3_ _5_ _3_ _§_ 3 5
603 <10 <10 <10 <10 <10 <10
614 <10 <10 <10 <10 <10 <10
615 <10 <10 <10 <10 <10 <10
616 <10 <10 <10 <10 <10 <10
617 <10 <10 <10 <10 <10 10
619 <10 <10 <10 <10 <10 <10
621 <10 <10 <10 <10 <10 <10
628 <10 <10 <10 <10 <10 <10
630 <10 <10 <10 <10 <10 <10
631 <10 10 <10 <10 <10 <10
632 <10 <10 <10 <10 <10 <10
633 10 <10 <10 <10 <10 <10
634 10 10 10 10 10 10
640 <10 <10 <10 <10 <10 <10
641 <10 <10 <10 <10 <10 <10
642 <10 <10 <10 <10 <10 <10
643 <10 <10 <10 <10 <10 <10
644 <10 <10 <10 <10 <10 <10
645 <10 <10 <10 <10 <10 <10
646 <10 <10 <10 <10 <10 <10
647 <10 <10 <10 <10 <10 <10
648 <10 <10 <10 <10 <10 <10
649 <10 <10 <10 <10 <10 <10
650 <10 <10 <10 <10 <10 <10
651 20 10 10 10 10 10
652 <10 <10 <10 <10 <10 <10
653 <10 <10 <10 <10 <10 <10
654 <10 <10 <10 <10 <10 <10
329
-------
TABLE A-18 ECHOVIRUS ANTIBODY TITERS BY HEMAGGLUTINATION INHIBITION
FROM 100 INDIVIDUAL PARTICIPANTS
Participant
I.D.
2
3
4
11
12
13
14
15
16
29
33
49
SO
74
75
76
83
84
97
101
102
141
143
145
146
157
158
159
162
165
166
172
177
178
181
182
196
199
206
207
213
214
226
227
228
229
231
233
241
242
244
260
Echovirus 6
Echovirus 13
Echovirus 19
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20
10
10
no
10
10
Echovirus 21
Period
1 5
no ~TO
10
Echovirus 25
Echovirus 29
1
10
To
i
10
10
10
10
10
10
20 10
10 40
'10
10 10
10
10
10 10
-10 10
10
10
10
10 10
10
10
10
10
10
10
10 10
10
10
10
10
20
20
(continued)
-------
TABLEA-18 (continued)
Participant
I.D.
284
290
294
312
324
325
372
374
386
391
441
443
445
446
457
464
465
500
504
510
603
614
615
616
617
619
621
628
630
631
632
633
634
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
Echovirus 6
Echovirus 13
Echovirus 19
Echovirus 21
~ 5
710
Period
1
716"
10
5
TIo
10
OO
10
Echovirus 25
Period
1 5
00 TTo
10
10
Period
3 5
<10 Oil
10
00
00
10
00
OO
00
00
Period
3 5
710 715
10
Period
3 5
715 7TO
00 00
10
10
10
10
00
10
00
10
710
00
00
10
00
Period
oTJ
10
715
OO
00
oo
00
oo
oo
10
oo
oo
oo
10
10
10
10
10
10
10
10
10
10
10
10
10
10
00
10
00
'.10
Echovirus
29
Period
1
710
OO
00
00
10
00
00
00
oo
00
10
00
oo
00
00
oo
00
oo
10
oo
Period
3
TIB
OO
00
oo
00
oo
00
1 n
-------
TABLE A-19 REOVIRUS ANTIBODY T1TERS BY HEMAGGLUTINATION INHIBITION
FROM 100 INDIVIDUAL PARTICIPANTS
Participant
I.D.
2
3
4
11
12
13
14
15
16
29
33
49
50
74
75
76
83
84
97
101
102
141
143
145
146
157
158
159
162
165
166
172
177
178
181
182
196
199
206
207
213
214
226
227
228
229
231
233
241
242
244
260
284
(continued)
Reovirus 1
Period
Reovirus 2
Period
Reovirus 3
Period
10
10
332
-------
TABLEA-19 (continued)
Participant Reovirus 1 Reovirus 2 Reovirus 3
I.D. PeriodPeriodPeriod
_1_ 5 1 5 1 5
290 <10 10 <10 <10 <10 <10
294 <10 <10 <10 <10 <10 <10
312 <10 <10 <10 <10 <10 <10
324 <10 <10 <10 <10 <10 <10
325 <10 <10 <10 <10 <10 <10
372 <10 <10 <10 <10 <10 <10
374 <10 <10 <10 <10 <10 <10
386 <10 <10 <10 <10 <10 <10
391 <10 <10 <10 <10 <10 <10
441 <10 <10 <10 <10 <10 <10
443 <10 <10 <10 <10 <10 <10
445 <10 <10 <10 <10 <10 <10
446 <10 <10 <10 <10 <10 <10
457 <10 <10 <10 <10 <10 <10
464 <10 <10 <10 <10 <10. <10
465 <10 <10 <10 <10 <10 <10
500 <10 <10 <10 <10 10 10
504 <10 <10 <10 <10 <10 <10
510 <10 <10 <10 <10 <10 <10
Period Period Period
_3_ _5_ _3_ _5_ 3 5_
603 <10 <10 <10 <10 <10 <10
614 <10 <10 <10 <10 <10 <10
615 <10 <10 <10 <10 <10 <10
616 <10 <10 <10 <10 <10 <10
617 <10 <10 <10 <10 <10 <10
619 <10 <10 <10 <10 <10 <10
621 <10 <10 <10 <10 <10 <10
628 <10 <10 <10 <10 <10 <10
630 <10 <10 <10 <10 <10 <10
631 <10 <10 <10 <10 <10 <10
632 <10 <10 <10 <10 <10 <10
633 <10 <10 <10 <10 <10 <10
634 <10 <10 <10 <10 <10 <10
640 <10 <10 <10 <10 <10 <10
641 <10 <10 <10 <10 <10 <10
642 <10 <10 <10 <10 <10 <10
643 <10 <10 <10 <10 <10 <10
644 <10 <10 <10 <10 <10 <10
645 <10 <10 <10 <10 <10 <10
646 <10 <10 <10 <10 <10 <10
647 <10 <10 <10 <10 <10 <10
648 <10 <10 <10 <10 <10 <10
649 <10 <10 <10 <10 <10 <10
650 <10 <10 <10 <10 <10 <10
651 <10 <10 <10 <10 <10 <10
652 <10 <10 <10 <10 <10 <10
653 <10 <10 <10 <10 <10 <10
654 <10 <10 <10 <10 <10 <10
333
-------
TABLE A-20 TRACE METAL CONCENTRATIONS IN BLOOD SAMPLES
FROM INDIVIDUAL PARTICIPANTS (us/100 ml)
Participant
I. P.
1
2
3
4
5
6
9
10
11
1Z
13
14
15
16
18
19
21
29
30
31 •
32
33
49
50
74
75
76
77
78
'83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
Cadmium
Period 1
0.093
0.092
0. 060
0.077
0.081
0.051
0. 060
0.040
0.051
0. 164
0. 075
0.058
0.324 .
0. 100
0.049
0.090
NSC
0.048
0.094
0.305
0.065
0.094
0. 063
0.051
0.075
0. 048
IS
IS
NSC
0.096
0. 155
0.068
- is
NSC
0. 182
0.040
0. 146
IS
0.033
0.040
IS
0.037
0.086
0. 033
0. 101
0. 051
Period 5
0. 105
0. 149
0. 049
0. 124
0. 042
0. 107
NSC
0. 064
0. 080
0. 061
0. 089
0. 180
0.089
0. 094
NSC
0. 063
NSC
NSC
0.077
0. 054
NSC
NSC
0. 097
0. 180
0. 087
0. 106
0. 132
NSC
NSC
0. 101
0. 166
0. 123
0.057
NSC
0. 082
0. 101
0. 092
0. 152
NSC
NSC
NSC
NSC
NSC
NSC
0.' 105
0. 149
110.8
70. 1
81. 8
106. 1
80.9
119.7
81.8
113.2
94.9
71.2
92.3
76.5
82.4
72.6
89.6
106. 0
NSC
111. 2
115.3
119.7
121.7
114.4
129.3
88.4
103.6
98. 1
100.3
116.7
NSC
129.3
116.4
112.9
105. 1
105. 5
NSC
108.6
99.9
121.7
76.6
99.9
102.6
111. 9
102. 1
96.7
122. 5
' 119. 1
Lead
Period 1
8. 1
5.4
12. 1
. 9.5
8.9
16.1
6.7
9.2
13.8
12.0
19.7
12.7
8.0 ;
9.0
12.0
6.4
.NSC
8.5
10.2
8.8
13.2
12.5
6.1
10.1
18.6
NSC
16.8
11.0
NSC
11.0 .
10.4
21.2
NSC
NSC
7.3 •
22.0
8.0
7.6
9.1
16.5
8.6
14.4
12.6
7.5
13.4
10.6
Period 2
10.8
NSC
NSC
9.9
11.5
11.8
10.3
8.9
13.9
13.0
21.6
15. 1
10.2
10. 2
12.6
8.9
NSC
7.3
9.7
7.0
9.8
17.3
7.5
11.6
14.6
9.7
NSC
NSC
NSC
11. 0
13.2
15.2
6.3
NSC
8.9
20.9
6.8
6.9
11.3
11.2
7.2
13.2
13.4
5.9
9.5
7.9
Period 5
12.0
9.7
9.7
7.9
7.9
10.8
NSC
7.6
8.4
9.5
20.4
17.9
18.8
14.3
NSC
9.4
NSC
NSC
10.5
8.0
NSC
NSC
8.6
11.4
13.2
8.6
13.5
NSC
NSC
14.0
10.5
13.8
5.5
NSC
11.8
19. 1
4.6
6.8
NSC
NSC
NSC
NSC
NSC
NSC
11.1
8. 1
Mercury
Period 1
0.23
0. 09
0.20
0.86
0.07
0. 57
0. 16
0. 82
0.47
0.65
0. 20
0.45
0.34
0. 57
0. 16
0.33
NSC
0.34
0.24
0.37
0.26
0. 57
0.69
0.33
0. 16
0. 16
0.40
0.20
NSC
0.45
0.44
0.26
0.27
NSC
1.07
0. 36
0.74
0.58
0.37
0.33
1. 13
<0.01
0.22
0.07
0.77
0. 19
Period 5
0.30
0. 18
0. 28
0. 16
0. 17
0.20
NSC
0. 12
1.70
0.31
0.38
0.31
0.28
0.60.
NSC
0.24
NSC
NSC
0. 13
0.26
NSC
NSC
0.48
0.05
0.24
0.30
0. 16
NSC
NSC
0. 19
0.01
0.28
0.31
NSC
0.40
0.42
0.39
0.38
NSC
NSC
NSC
NSC
NSC
NSC
0.35
0.44
Zinc
Period 1
496
407
701
654
509
393
423
675
489
421
736
612
649
503
476
538
NSC
579
435
482
675
728
530
556
526
649
503
632
NSC
566
619
660
455
NSC
544
719
877
737
596
632
570
455
649
658
• 737
579
(continued)
-------
TABLE A-20 (continued)
Participant
I. D.
143
144
145
146
157
158
159
160
16Z
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
Cadmium
Peri- 1
IS
0. 068
0. 103
0. 044
0. 137
0.015
NSC
NSC
0. 296
0.036
0. 191
NSC
NSC
NSC
0. 129
0.215
0. 152
0.-122
IS
0. 075
NSC
0.051
0. 077
0. 061
0. 063
NSC
0. 067
NSC
0. 113
0.030
0. 037
0. 129
NSC
0. 109
0. 103
IS
0.114
0.069
0. 081
IS
0. 137
0. 133
0.054
IS
0. Ill
NSC
0. 071
Period 5
0.044
0.045
0. 105
0. 068
0. 277
0.085
0. 124
0. 130
0.094
0.042
0. 135
NSC
0. 073
0. Ill
0. 188
0. 154
0. 178
0.092
0. 070
0.102
0.070
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0.089
NSC
0.092
NSC
NSC
0.220
0. 136
0.223
0. 125
0.094
0.099
0. 106
0. 151
0. 151
0.035
NSC
0. 068
0. 105
0. 073
103.9
129. 5
105.8
93.3
88.6
122.8
NSC
NSC
170.4
94.2
• 99. 5
80.5
NSC
NSC
114.9
93.3
95. 5
120.3
77.8
95.2
NSC
88.3
93.6
91. 5
92.3
NSC
98.6
NSC
65. 1
81.0
139.4
66.2
NSC
80. 2
116.8
91.2
109.0
108.6
93.3
80.2
96.2
50.6
108. 4
127.0
140.4
NSC
125.3
Lead
Period 1
11.6
7.6
18.8
5.6
17.4
7.2
NSC
NSC
5.2
9.0
5.2
5.6
NSC
NSC
10. 1
9.9
6.3
9.5
30.6
14.7
NSC
13.9
4.1
5.8
7.9
NSC
7.3
NSC
16.7
10.5
21.4
7.7
NSC
13.9
8.5
16.3
10. 1
11.4
8.0
6.4
16.9
7.8
13.2
14.7
8.6
NSC
5.0
Period 2
5.6
7.2
15.5
5.7
16.3
6.8
NSC
NSC
8.0
11.3
5.4
NSC
10.7
10.7
13.8
9.4
7.0
10.4
17.2
7.2
11.7
11. 1
5.3
5.5
8.3
13.2
6.8
NSC
11.3
13.5
13.2
12.2
11. 1
9.5
8.0
9.8
7.0
10.8
10.4
5.7
12.9
8.5
22.8
NSC
7.5
NSC
9.0
Period 5
8.7
5.0
12.9
6.7
16.6
6.8
6.8
7.5
9.2
8.1
9.9
NSC
8.0
7.7
13.0
10.6
8.6
11.4
14.8
8.6
9.7
NSC
NSC
NSC
NSC
NSC
NSC
NSC
9.2
NSC
15.8
NSC
NSC
11. 1
8.6
9.2
9.8
9.2
8.7
6.4
12.0
7.4
15.8
NSC
10. 0
9.9
8. 5
Mercury
Period 1
1. 41
0. 57
1.43
0. 71
0.28
0. 18
NSC
NSC
0.69
0.86
0.22
0. 71
NSC
NSC
0. 22
0.30
0. 11
0. 35
0.61
0. 13
NSC
0.27
0.30
0.22
0. 09
NSC
0.65
NSC
1.48
0.04
0. 51
0. 05
NSC
0.65
0.84
0.34
0.21
0.37
0.24
0. 22
0.22
0.65
0.49
8. 11
1. 02
NSC
0. 04
Period 5
0.30
0. 39
0.73
0. 37
0.32
0.36
0.25
0.22
0. 18
0.38
0. 18
NSC
<0. 01
0. 11
0. (2
0.05
0.56
0. 14
0.45
0. 26
0.06
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0. 50
NSC
0.37
NSC
NSC
0. 49
0.23
0.25
0. 23
<0.01
0. 10
0.32
0. 07
0.36
0. 30
NSC
0. 82
0.03
0.30
Zinc
711
833
591
641
566
509
NSC
NSC
939
798
448
660
NSC
NSC
490'
522
614
523
614
702
NSC
606
484
503
553
NSC
373
NSC
746
667
489
537
NSC
579
711
728
592
723
540
649
482
428
708
503
606
NSC
537
(continued)
-------
TABLE A-20 (continued)
Participant
I. D.
235
241
242
Z43
244
254
Z55
Z56
257
258
260
262
263
266
267
268
273
275
276
278-
Z84
290
293
294
295
30Z
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
348
(continued)
Cadmium
Period 1
NSC
0.111
0. 026
0.034
0.058
0.090
NSC
0. 101
0.076
0. 1Z9
0. 103
0.055
0. 071
0. 09Z
IS
0. 122
0. 210
0.054
0.091
0.072
0. 079
0.081
0.323
IS
0. 094
0. 116
0. 148
0.050
0.068
0.150
NSC
0. 123
IS
0.295
0.046
0. 155
0.087
0.040
0.054
IS
0. 142
NSC
NSC
NSC
0. 131
0. 103
Period 5
0.072
0.059
0.051
0.086
0.080
NSC
NSC
NSC
NSC
NSC
0.057
0. 128
0.136
0.044
0. Ill
0.080
0. 191
0.042
0.330
0.089
NSC
0.089
NSC
NSC
NSC
0.066
0.232
0.073
0. 101
0.054
NSC
0. 105
0. 123
0. 146
0. 135
0.249
0.049
0.072
0.064
0.064
0.095
0.086
0. 180
NSC
0. 123
0.059
NSC
93.3
98.6
104.6
105. 1
125.6
NSC
96.4
83. 1
84.4
114. 9
124.0
123. 8
97.4
86. 1
50.6
103. 9
93.6
96.2
109.7
101. 1
173.8
102.6
100.4
72.9
86.4
82.8
90.4
110.8
63.9
NSC
62.1
108.2
105. 8
97.0
123.4
80.6
115.7
90.7
153.3
95.7
NSC
NSC
NSC
96.4
99. 2
Lead
Period 1
NSC
4.8
13.7
4.8
6.3
15.3
NSC
7.8
13.4
12. 1
13.8
15.2
13.0
11.2
8.9
7.5
11.5
8.5
6. 1
10.1
15.6
10.9
19.9
25. 1
10.3
11.0
6.7
21.8
20.9
7.8
NSC
10.7
7.8
7.2
11.2
17.8
11.3
15.7
17.4
10.
11.8
NSC
NSC
NSC
12.6
6.9
Period 2
NSC
5.9
14.7
6.8
8.0
NSC
10.2
7.0
14.8
11.0
11.2
13.4
13.6
9.8
6.9
9.0
12. 1
5.5
5.5
13.3
16.7
6.3
15.2
17.2
NSC
10.2
8.1
27.2
13.9
7.7
10.4
7.5
7.8
8.3
13.5
25.8
13.2
13.3
13.0
NSC
18.0
NSC
12.0
7. 5
10.4
NSC
Period 5
9.0
8.4
16.0
6.5
8.6
NSC
NSC
NSC
NSC
NSC
9.0
14.9
12.6
11.8
11.2
11.6
12.5
6.5
8.0
9.5
NSC
6.2
NSC
NSC
NSC
9.9
11.8
14.4
17.4
NSC
9.2
6.2
6.2
6.8
11.2
14.2
12.3
9. 1
10.7
11. 1
16.0
9.5
7.4
NSC
8.9
5.6
Me rcury
Pe riod 1
NSC
0. 16
0.68
0.61
0.68
0.66
NSC
0.43
0. 19
0.33
0.61
0.95
0.64
2.40
1.71
0. 12
0.68
0.20
0.31
0. 12
0.06
1.28
0. 09
0.28
0.18
0. 15
0.07
0.80
0.44
0. 50
NSC
0.82
0.39
0.45
1.07
0.47
0.54
0.69
0.74
1.19
1. 15
NSC
NSC
.NSC
0.54
0.46
Period 5
0.09
0. 18
0. 27
0. 11
0. 13
NSC
NSC
NSC
NSC
NSC
0. 16
0.37
0.24
0.44
0.78
0.09
0.27
0. 08
0.04
0.21
NSC
0.39
NSC
NSC
NSC
0. 50
0.59
0. 10
0. 14
NSC
<0. 01
0.38
0. 17
0.27
0.39
0.09
0. 19
0.08
0.26
0. 10
0.46
0.08
0.05
NSC
0.37
0. 15
Zinc
NSC
684
540
428
441
641
NSC
523
570
647
714
614
570
572
798
708
693
517
469
414
628
675
452
519
667
670
740
638
579
386
NSC
638
698
554
584
647
439
605
612
451
541
NSC
NSC
NSC
562
564
-------
TABLE A-20 (continued)
Participant
I. D.
349
350
353
354
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
Cadmium
Period 1
0.091
0. 124
0. 087
0. 118
0.092
IS
0.053
IS
IS
NSC
0. 099
0.099
IS
IS
0.098
0.040
IS
NSC
0. 159
NSC
0.092
0. 078
0.031
0. 129
0.044
0.047
0.044
0.030
IS
0.060
0.044
IS
0. 073
IS
0. 041
0. 048
0. 143
0. 066
0.062
0. 108
0.020
0.052
0.058
0.070
0. 102
Period 5
NSC
NSC
0. 185
0. 127
0. 035
0. 078
0. 138
0. 030
0. 197
0. 044
0. 106
0.042
0. 130
0.211
0. 086
0.054
IS
0. 074
0. 125
0.204
0.205
NSC
NSC
0.044
0.042
0. 121
0. 184
0.088
0.087
0. 133
NSC .
NSC
NSC
NSC
NSC
0. 078
0. 118
0. 231
NSC
0. 082
0.094
0. 101
0.089
0.068
NSC
55.6
98.6
95.4
99. 2
120. 0
110. 0
130. 1
116. 4
94. 8
NSC
95.7
125.6
98.6
81.4
111. 4
125. 1
125.6
NSC
97. 0
NSC
64.2
119.9
51.6
73.3
94. 8
138.6
117. 2
94. 0
110. 0
97. 0
109. 7
85.7
118.7
121. 9
155.0
104.6
124. 1
85.7
96.4
128.2
92.3
88.2
128. 0
100. 5
101. 7
Lead
Period 1
7.4
9.8
13.9
17. 1
11.9
13.0
13.7
1.9
7. 1
NSC
17.5
8.9
9.9
5.6
6.0
7.7
9.7
NSC
16.8
NSC
22.4
14.1
21. 5
11.8
8.6
8.3
12.7
13.8
18. 1
9.8
8.3
13.4
13.0
12. 1
10.6
10. 1
9.4
15.3
15.9
13. 5
14.4
11.4
7.6
11.8
15.2
Period 2
7.0
5. 1
9.0
9. 1
19.7
12.0
7.4
7.6
9.1
NSC
11. 0
8.2
8.0
5.3
8.5
6.8
9.7
NSC
18 9
NSC
NSC
NSC
NSC
12.8
4.1
9.7
13. 1
10.2
NSC
8.6
9.4
7.4
9.7
NSC
5.4
7.8
7.2
10.0
12.6
16.3
7.6
8.2
6.1
12.0
12.2
Period 5
NSC
NSC
8.6
11. 1
14.9
12.7
9.4
3.8
6.8
12.0
11.1
10.3
10.4
6.3
9.5
10.3
9.9
5.5
18.8
10.5
18.0
NSC
NSC
10.2
9.6
9.9
12.9
9.9
14. 1
12.3
NSC
NSC
NSC
NSC
NSC
5.8
8.0
12.3
NSC
8.0
9.2
5.6
5. 1
12. 5
NSC
Mercury
Period 1
0.35
0. 11
0. 24
0. 24
0.38
6.36
0. 06
1.25
0.38
NSC
0.05
0. 22
0.52
1. 20
0.92
0. 07
0.31
NSC
0.38
NSC
0.70
0.28
0.28
0.22
0.42
0.31
0. 76
0.74
2. 17
0. 19
0. 49
0.20
1. 51
0.35
0.80
0.44
0.77
1.04
0.01
0.60
0. 58
0.21
0.24
0.24
1. 18
Period 5
NSC
NSC
0. 16
0.34
0.32
0. 22
0. 21
0.74
0. 06
0. 24
0.08
0. 17
0.22
0. 32
0.03
0. 18
IS
0. 17
0.42
0. 05
0. 15
NSC
NSC
0.20
0. 43
0.28
0.21
0.36
0. 03
0. 09
NSC
NSC
NSC
NSC
NSC
0.38
0.48
0.82
NSC
0. 55
0. 27
0. 23
0. 15
0. 20
NSC
Zinc
565
544
693
572
474
494
604
456
425
NSC
638
613
612
813
548
477
605
NSC
658
NSC
598
689
526
746
632
466
585
559
783
612
536
736
404
456
725
638
740
695
623
690
571
596
553
638
690
(continued)
-------
TABLE A-20 (continued)
Participant
1. D.
413
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467 •
500
501
502
503
504
505
506
507
(continued)
Cadmium
Period 1
0. 101
NSC
0. 174
IS
120
\076
0. 125
0.044
0.032
0.094
0.208
0.068
0. Ill
0.312
NSC
IS
IS
0.040
IS
0.059
IS
IS
IS
NSC
IS
0.051
0.090
0.078
0.047
0. 040
0.029
0.060
IS
IS
0. 121
0.030
0.049
IS
0.045
0.099
0.090
0.070
NSC
IS
IS
Period 5
NSC
0.066
0. 155
0.063
0. 197
0.040
0. 125
NSC
0.040
0. 048
0.048
0.067
0.062
0.330
NSC
NSC
0.080
0. 095
0. 133
0. 143
IS
0.230
0.063
NSC
NSC
NSC
NSC
0.054
0. 057
0. 154
0.072
0. 101
0.087
0. 171
0. 061
NSC
NSC
0. 216
NSC
NSC
NSC
0.086
0. 021
0. 072
0.062
123. 8
NSC
90. 8
96.4
98. 5
86. 1
92.9
96.4
110. 5
88. 2
96.4
83.2
115. 9
108.7
NSC
146.7
126.2
127.7
130. 8
81.6
124.2
79.7
97.6
NSC
98.0
101.0
172.2
129. 2
139.8
115.7
104.6
79.2
105. 5
89.7
37. 5
77.2
95. 0
147.8
98. 9
63.9
107.3
89.7
NSC
141.4
100.3
Lead
Period 1
9.2
NSC
17. 5
13. 5
26. 1
20.2
18.9
15. 9
10.3
10.3
7.8
8.9
11.6
11.9
NSC
8.8
6.0
12.5
9.9
6.7
12.0
25.4
9.5
NSC
11.8
13.4
12.6
20.4
9.3
9.9
7.6
7. 5
6.4
10. 1
10.
12. 5
13.6
17.2
14. 1
8.6
12.0
8.7
NSC
13.9
12.6
Period 2
7.3
NSC
14. 5
15.0
29. 0
25.8
16.5
11. 1
7.2
11.0
6.6
8.5
12. 5
8.9
NSC
13. 5
5.9
8.2
8.5
9.2
11.9
23. 5
NSC
NSC
10. 5
11.9
11. 5
18. 1
9.0
10. 1
9.9
5.7
7.8
18.9
14. 5
6.5
12.7
NSC
NSC
10. 1
11.7
9.1
NSC
14.3
7.7
Period 5
NSC
9.9
10.3
13.3
18. 4
19.3
13.5
NSC
7.6
10.9
7.6
8.0
8.0
8.6
NSC
NSC
11. 1
'7.4
9.9
9.0
9.9
21.4
8.3
NSC
NSC
NSC
NSC
14.0
8.0
7.9
7.2
4.8
3.7
12.2
11.0
NSC
NSC
11.4
NSC
NSC
NSC
8.2
12.3
7.6
9.5
Mercury
Period 1
0.24
NSC
0. 01
0. 16
0. 44
0. 20
0.06
0.62
0. 36
0.64
0.43
0.61
0.28
0.36
NSC
0.88
0. 51
0.29
0. 16
0.28
0. 21
0. 17
8. 14
NSC
0. 51
0.65
0.71
0.27
0. 51
0.48
0. 15
0. 15
0.67
0. 19
0. 11
0. 18
0. 16
0.48
0.30
0.99
0.07
0.29
NSC
0.33
0.61
Period 5
NSC
0.23
0. 18
0. 15
0. 13
0. 25
0. 28
NSC
0. 53
0.45
0.09
0. 10
0. 16
0. 16
NSC
NSC
0.41
0.09
0.43
0. 10
IS
0.34
0.09
NSC
NSC
NSC
NSC
0.39
0. 16
0.08
0. 19
0.23
0.37
0.28
.0. 17
NSC
NSC
0. 12
NSC
NSC
NSC
0. 12
0. 12
0. 12
0. 13
632
NSC
391
323
622
664
585
502
630
638
439
537
687
681
NSC
658
711
488
693
638
491
640
373
NSC
529
478
612
784
701
351
460
723
647
640
. 526
613
587
719
664
720
562
417
NSC
597
675
-------
TABLE A-20 (continued)
Participant
I. D.
508
509
510
511
512
Cadmium
0. 169
0.057
0.050
NSC
NSC
0.251
0.064
0.078
NSC
0. 091
119.4
149. 2
101. 0
NSC
NSC
Lead
Period 1
12.9
10. 1
14. 1
NSC
NSC
Period 2
14.5
10.2
15.5
NSC
15.3
Period 5
12. 1
10.5
18.0
NSC
10.5
Period 1
0. 51
0.80
0. 56
NSC
NSC
Mercury
Period 5
0.27
1.06
0.28
NSC
0.09
Zinc
605
748
660
NSC
NSC
-------
TABLE A-21
TRACE METAL CONCENTRATIONS IN FECES SAMPLES
FROM INDIVIDUAL PARTICIPANTS (Mg/g)
Participant
I.D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
21
29
30
31
32
33
49
50
70
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
(continued)
Cadmium
Period 1
0. 138
0. 082
0. 134
0. 119
0. 154
0. 1E3
0. 243
IS
0. 112
0. 122
0. 307
0. 061
0. 125
0. 184
0. 163
0. 265
0. 155
0. 295
0.355
0. 121
0. 131
0. 130
NSC
0. 086
NSC
0. 164
0.051
NSC
0. 207
IS
0.090
0. 161
0.087
IS
0. 188
0. 148
0.223
0.373
0. 17Z
0.331
0. 210
0.040
0. 240
0. 154
0. 097
0. 180
Period 5
0. 180
0. 155
0. 086
0. 069
0. Z08
0. 235
0. 148
0. 210
0. 114
0. 155
0. 030
0. 061
NSC
0. 193
0. 511
0. 127
NSC
0. 210
0. 098
0. 136
0. 029
0. 144
0. 083
0. 121
NSC
0. 054
0. 150
0. 148
0. 130
0. 200
0. 261
0. 038
0. 073
0. 318
0. 157
0. 116
0. 054
0. 106
0. 269
NSC
NSC
NSC
NSC
NSC
0. 146
0. 340
Lead
11.3*
11.9
17. 1
5.7
14. 2*
5.2
15.4
16. 0
12. 1
12. 5
18.9*
. 1*
.3
16. 1
7. 2
9.8
6.0
17.6
18.6
31.
18.
25.3
27.3*
7. 0
NSC
12.9*
NSC
22.4
5.3
NSC
14. 0
32. 7
10.2
13. 9
10.6
16. 0
Z3.9
24.6.
27.8
28.8*
28.8*
15. 7
14.7
17. 7
18.3
22.2
13.0
13.2
Period 1
0. 94*
1. 75*
1. 04
0. 93
1. 15*
1.09
0.86
0. 83
1. 09
0.88
3.69*
0. 63
0. 20
0. 43
0,48
2.04
0. 90
2.38*
2. 07
1.70
2. 08*
1. 11*
NSC
0. 58*
NSC
2. 15
1. 16
NSC
0. 75
1. 14
0. 53
0. 82
0. 29
0.68
0. 80
1. 27
0.89
0.60*
1. 11*
1.46
2. 18
1. 10
2.49
1. 56
0.85
1.29
Period 5
0.32
0. 56
0.77
0. 15
0.66
0. 51
1. 20
0. 45
1. 10
0.38
0.69
0.35
NSC
0.33
0. 73
1.49
NSC
1.53
0.98
0.61
1. 29
0.72
0.68
0.38
NSC
0. 24
0. 55
1. 78
1. 24
1. 06
0. 94
0. 62
0.55
IS
0. 56
IS
0.37
0. 40
1. 16
NSC
NSC
NSC
NSC
NSC
0. 53
IS
Mercury
Period 1
0. 17
0. 10
0. 05
0.04
0. 06*
<0. 04
0. 11
0. 19
0. 12
0. 26
0.64
0. 41
0. Z2
0. 20*
c. :3*
0. 27
0. 04
0. 16
<0. 04
0. 14
<0. 04
<0. 04
NSC
0. 20*
<0. 04
0. 09
<0. 04
NSC
<0. 04
0.06*
<0. 04
<0. 04
0. 14
0. 09
<0. 04
0.04
<0.04
0.35
0. 17*
<0. 04*
0. 14
-------
TABLE A-21 (continued)
Participant
I.D.
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
177
178.
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
Cadmium
Period 1
0.246
0. 161
0. 184
0. 161
0.214
0.201
0.055
0. 135
IS
0. 176
0. 666
0. 180
0.258
0. 173
IS
0. 074
0.030
0. 087
0. 142
0. 105
0.213
0.318
0.299
0. 131
0. 172
IS
0. 276
0. 298
0. 192
IS
0.358
0. 080
0. 085
0. 127
0. 184
0. 266
0. 204
0. 237
0.079
0. 064
0. 067
0. 169
0. 112
0.205
NSC
0. 103
Period 5
0.023
0. 029
NSC
0. 079
0. 140
0. 070
<0. 014
0. 072
0. 086
0. 042
0. 038
0. 193
NSC
0.488
0.034
0. 057
0. 114
<0. 014
0. 024
0. 075
0. 127
0. 150
NSC
NSC
NSC
NSC
NSC
NSC
NSC
<0.014
0. 091
0.035
NSC
NSC
0.098
0.065
0. 053
0. 188
0. 121
0. 173
0. 106
0. 099
0. 106
0. 442
0. Z88
0. 094
Lead
14.2*
9.6*
15.5
21.3
21.3
11.8
16. 1
6.5
5.3
10. 9
26.9
18.2
33.
17.
11.
. 5
.2
.2
7. 5*
27. 0
23.0
26.0
11.7
17.5*
8.2
21.3*
14.2*
18.2
11.9
16.8
14. 1
22.6*
14.8
14.5
19. 1
20.4
23.4
32.5
26.0
13.1
21.5*
19.6
24.3
23.2
15.3
14.4
26.5
NSC
26.9
Period 1
1. 22*
0. 46*
1/16
1. 93*
1. 12
0.38
0.6Z
0. 19
0.49
0.34
I. 27
0. 80
0.85
1.02
0. 29
0.33
0. 76
1. 18
2. 85
0. 17
0.38*
0. 70
1. 54*
0. 54*
0. 87
0. 46
1.36
0.66
1.67*
1. 40
1. 04
2.39
0. 82 .
1. 34
<2. 05*
2. 55
2.68
3. 12*
0. 12
0. 93*
1. 21
1. 06
0. 28
•1. 98*
NSC
0.39
Period 5
1. 25
IS
NSC
0.31
0. 70
0.79
3. 72
1.46
1. 99
0. 21
0.49
0. 46
NSC
0.73
0. 50
0.47
0.61
0. 98
0. 59
0.24
0. 53
1. 10
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0.31
0. 93
0.48
NSC
NSC
0.24
0.43
0.32
0.38
0. 41
0.35
0. 81
0. 13
0. 18
0. 79
11.3
0.47
Mercury
Period 1
0. 53
0. 05*
0. 20
0.30
0. 15
0. 13
0. 29
0. 05
0. 51
0.31
0.63
0.08
0.88
0.22
1. 57*
0.09
<0. 04
<0. 04
0. 19
<0. 04
0. 17*
<0. 04
0. 07*
0. 11*
0. 18
0.07
<0. 04
0. 11
•=0. 04
0. 19
<0. 04
0. 19
0. 13
<0. 04
0. 12*
0.29
0.06
0. 40
<0.04
<0. 04
0. 26
0. 17
0.49
0. 12*
NSC
0. 12
Period 5
0. 16
IS
NSC
0. 04
0. 06
0. 17
0.07
0.04
< 0. 04
0.07
0.61
< 0. 04
NSC
<0.04
0.08
<0. 04
0. 06
0. 09
0.22
<0. 04
0.21
<0. 04
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0.09
<0. 04
0. 05
NSC
NSC
0. 27
0.05
<0. 04
<0. 04
<0. 04
<0.04
0.42
0.22
0.36
0. 12
<0. 04
0. 15
Zinc
t'eriod 1
62*
49*
55
117
97
130
83
61
1298*
96
183
104
88
36
53
• 54
130
125
126
126
120*
91
186*
60*
130*
51
129
121
147*
85
128
145
73
149
188*
124
68
171*
73
98
104
153
96
175*
NSC
138
(continued)
-------
TABLE A-21 (continued)
Participant
I.D.
232
233
235
239
241
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
295
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
Cadmium
Period 1
0.057
0.052
0. 135
0. 147
0.086
0. 151
0. 112
0. 169
0. 134
0.301
0. 106
0. 141
0.049
0.086
0.039
0.326
0.163
0. 115
0. 168
0. 112
0. 170
0.062
NSC
0.085
0. 114
NSC
NSC
NSC
0.048
IS
0. 131
0. 143
0.385
0. 139
0.085
0. 106
0. 114
0. 233
0. 118
0. 141
0. 140
0. 176
0. 160
NSC
0. 172
NSC
0. 056
Period 5
0. 196
0. 254
0. 149
NSC
0. 115
0. 051
NSC
0. 113
NSC
NSC
NSC
NSC
NSC
0. 244
0. 034
0. 058
0. 078
0. 090
0. 060
0. 059
0. 237
0. 178
0. 051
0. 014
0. 035
NSC
0. 125
NSC
0. 098
0; 057
0. 052
0. 267
NSC
0.253
0. 204
0. 101
0. 049
0.093
IS
0. 133
0. 026
0. 026
0. 069
0. 261
0. 036
0. 016
0. 045
Lead
27.2
24.0
12.8*
27. 1
12.3
17.4
14.2
22.4*
12.2
20.3*
22.7
NSC
18.2
19.3
4.4*
13.6
10.7
13.2
19.9
6.0
17.5
15.2
NSC
12.0
9.6
NSC
NSC
NSC
17. 4
17. 0
8.8
4.7
13.3
9.7
10.0
8. 1
13.3
10. 1
8.7
16.7
15.9
10.7
17.7*
NSC
14.7
NSC
15.2
Period 1
1. 14
2.39
2.85*
0.62
0.60
1.81
0.48
1. 26*
0.60
11.98*
3.32*
NSC
2.76
0.40
1.65*
2.36
0.62
1.37*
0.90
0.25
0.54
0.98
NSC
0.51
2.01
NSC
NSC
NSC
0.79
0.37
1.06
0.28
0.60
1. 99
1. 19
0. 56
1. 13
0.69
0. 59
0.70
1.01
0.88
1.01*
NSC
2. 06
NSC
0.42
Period 5
1.28
0.61
0.63
NSC
0.67
0.61
NSC
0.75
NSC
NSC
NSC
NSC
NSC
0. 18
0. 52
0.91
0.56
0.85
1.06
0.45
0. 51
0.93
0. 18
0.84
0. 59
NSC
0.75
NSC
1. 02
1. 51
0.72
0.48
NSC
0.91
0. 5.9
1. 14
0.45
1.57
0.73
0. 15
0. 27
0.56
0.41
0.61
1.09
0. 73
0. 59
Mercury
Period 1
<0. 04
0.25
< 0. 04
0. 07
0.09
0.06
0. 06
0. 26*
0.05
0.07
0. 13*
NSC
0.09
< 0.04
0. 16*
< 0.04
< 0.04
0.38
< 0. 04
< 0. 04
< 0.04
0.06
NSC
0. 10
0.39
NSC
NSC
NSC
< 0. 04
NSC
< 0.04
< 0. 04
<0.04
<0. 04
<0.04*
<0. 04
3. 51*
0.06
0.20
0.05
<0.04
0.05
< 0. 04*
NSC;
0. 18
NSC
<0. 04
Period 5
< 0. 04
0. 16
< 0. 04
NSC
0. 12
0.09
NSC
0. 10
NSC
NSC
NSC
NSC
NSC
< 0. 04
< 0.04
< 0. 04
< 0.04
0. 13
< 1. 14
IS
< 0. 04
< 0. 04
< 0. 04
0. 05
0.31
NSC
' 0. 08
NSC
0. 09
0. 11
0. 05
0.09
NSC
< 0. 04
< 0. 08
0. 08
0.23
0.05
IS
< 0. 04
0.04
<0. 04
<0.04
0.31
<0. 04
0.04
0. 07
Zinc
Period 1
74
138
89*
95
83
74
111*
73*
82
74*
92*
NSC
79
• 70
71*
151
58
179*
49
65
96
110
NSC
69
103
NSC
NSC
NSC
110
82
74
50
85
75
97
48
66
38
99
82
62
74
69*
NSC
59
NSC
106
(continued)
-------
TABLE A-21 (continued)
Participant
I.D.
343
348
349
350
353
354
355
356
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
(continued)
Cadmium
Period 1
0. 110
0. 118
0.063
0. 130
0. 176
0. 106
0.094
0. 258
0. 150
IS
0. 062
0. 147
0. 202
0.331
NSC
0. 144
IS
NSC
0. 137
NSC
NSC
0. 382
0. 076
0. 090
0.210
NSC
0. 058
0.237
0. 080
0. 260
0.089
0. 209
0. 052
0. 190
0. 184
0. 255
0.223
0. 060
0. 155
0. 106
IS
IS
0. 187
0. 191
0. 048
NSC
Period 5
0. 098
NSC
0.030
0. 109
NSC
0.226
0. 217
0.306
0.026
<0.014
NSC
NSC
0. 283
0. 117
NSC
0. 113
0. 131
0.230.
0. 102
0. 119
0. 110
NSC
0. 134
NSC
NSC
0. 109
0. 184
NSC
0. 014
0.049
0. 048
0.019
0.017
NSC
NSC
NSC
NSC
0. 049
0. 165
0. 113
NSC
0. 027
0. 174
0. 254
0. 065
0. 219
Lead
8.4
10. Z
26.9
12. 3*
11.4
17.3
11.3*
9.3*
29.4*
26.0
14.6*
19.3
7.3
18. 5
NSC
18.8
18.1
NSC
1Z.9
NSC
NSC
24.6
21.6
16.5
8.9*
NSC
5.3
29.6
26. 1
32.6*
9.7
47.3
32.4
9. 1
14.3
12. 4
12.5
26.0
12.7
30.8
19.8*
27.9
14.0
21.6
18.8
NSC
Period 1
0.63
0.43
0. 57
0. 51*
0.67
1. 18
0. 88
2. 43*
0. 52*
0. 60
0. 88*
0. 91
0. 46
1. 03
NSC
0. 74
0.66
NSC
1.32
NSC
NSC
2. 43
1. 10
3. 06*
0. 55*
NSC
0. 27
1. 72*
1.79
1. 17*
1.69
2.89
2.20
0.39
1.37
4. 91*
3.04
1.27
0. 95
1.68 .
2. 19*
1. 56
1. 54
1. 53
0. 82
NSC
Period 5
< 0. 10
NSC
IS
<0. 10
NSC
0. 56
0. 15
0.67
0. 73
0.48
NSC
NSC
0. 85
2.48
NSC
0.85
0. 95
1.88
0.30
0.83
0.45
NSC
0.24
NSC
NSC
0.43
0. 87
NSC
0. 54
0.35
1.28
0.32
0. 72
NSC
NSC
NSC
NSC
0. 98
0. 35
0. 96
NSC
0.45
0. 17
0.35
0.24
0. 19
Mercury
Period 1
0.07
<0.04
0. 13
0. 06*
0. 24
0.49
0:08
<0.04
0.22
0. 17
<0.04*
< 0. 04*
<0. 04
0.05
NSC
0.26
1. 04*
NSC
< 0.04
NSC
NSC
<0. 04
<0. 04
0. 13*
0.05*
NSC
0. 10
0. 14*
0. 48
< 0. 04
<0. 04
0. 16
<0. 04
0.46
0. 11
<0. 04
0. 31
0.43
0. 23
0.62
0. 13
NSC
0.21
0. 13
0. 07
NSC
Period 5
0.05
NSC
IS
< 0. 04
NSC
0.06
0. 04
0. 28
0.35
0. 21
NSC
NSC
<0.04
< 0. 04
NSC
0. 11
0. 05
0.70
<0. 04
<0. 04
0.31
NSC
< 0. 04
NSC
NSC
0. 04
0. 10
NSC
0.08
0. 13
< 0. 04
0. 05
<0. 04
NSC
NSC
NSC
NSC
0.37
0.28
0. 59
NSC
0. 19
<0.04
0. 17
0. 15
0. 14
Zinc
Period 1
102
73
95
86*
72
93
52*
44*
114*
79
64
88
110
• 119
NSC
96
130
NSC
107
NSC
NSC
156
70
166*
56*
NSC
52
167*
149
85*
78
187
195
60
72
67*
100
103
103
181
205*
289*
106
69
96
NSC
-------
Participant
I.D.
412
413
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
W 449
£ 450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
512
(continued)
Period
0. 272
NSC
NSC
0. 195
0. 087
NSC
0. 089
0. 155
IS
0.226
0. 036
0.217
0. 090
0.070
0. 109
0.089
0. 114
0.094
0.097
0.255
0. 144
0. 281
0. 211
NSC
0. 075
0.239
0.052
0. 181
0. 076
0. 164
0. 175
0. 134
0. 149
0.201
0. 119
IS
NSC
0.047
0. 126
0. 091
0. 170
NSC
NSC
0. 240
0.226
NSC
NSC
0.099
0.088
NSC
Cadmium
1 Period 5
NSC
NSC
NSC
NSC
0.200
NSC
0. 133
0. 057
< 0. 014
0. 126
0. 102
0. 243
0. 067
0. 155
0. 126
0. 059
NSC
0. 06.8
0. 174
0. 166
0. 184
NSC
0. 127
NSC
NSC
NSC
NSC
NSC
0. 033
0. 057
<0.014
0.078
0. 075
NSC
0. 029
0. 108
NSC
NSC
NSC
NSC
NSC
NSC
0. 151
0. 102
0. 187
NSC
0.370
0.336
0. 136
0.245
TABLE A-21
Copper
Period 1
15.0*
NSC
NSC
12.8*
14.6
NSC
30.0
8.7
8.2
12.9
15.4
10. 1
3.7
2.8
11.7
10.8
9.7
17. 1
9.7
14.2
12.2
10.6
9. 1
NSC
7.7
11.2
17.2
13.8
14.0
11.2
10.0
20.8
25.7
35.8
13.8
19.1
NSC
3.1*
24.6
13.0
13.6
NSC
NSC
20.8
21.7
NSC
NSC
22. 1
7.2
NSC
(continued)
Lead
Period 1
0. 82*
NSC
NSC
3.06*
1.94
NSC
2. 19
0. 90
0. 82
0.87
1. 56
0.48
0.33
1.04
0.86
1.22
0.61
0. 92
0. 81
1. 14
0.71
2. 19
1.37
NSC
0.69
0.70
1.39
2.41
0. 76
0. 55*
0. 97
0.89
1. 12*
0.84*
0.83
1.36
NSC
0. 18*
Z. 72*
0. 92
0.71
NSC
NSC
1.97
1.62
NSC
NSC
1. 54*
1.47
NSC
Period 5
NSC
NSC
NSC
NSC
0.29
NSC
0. 54
0. 38
0. 61
0.64
1. 84
0.29
1.28
1.63
0. 16
0.45
NSC
2.24
IS
0.84
0.46
NSC
0.89
NSC
NSC
NSC
NSC
NSC
0.62
1. 23
1. 25
0.27
0.29
NSC
1.20
1.05
NSC
NSC
NSC
NSC
NSC
NSC
1. 04
1.58
0. 19
NSC
0. 53
0.97
0. 15
0.68
Mercury
Period 1
0.07*
NSC
NSC
< 0.04
< 0. 04
NSC
0.20
0. 10
10.39*
0. 15
< 0.04
0. 14
0.09
0.06
0. 16
< 0.04
0. 15
0. 13
< 0. 04
0.08
0. 18
< 0.04
0.43
NSC
< 0.04
0.33
0. 13
0.29
0. 04
0. 15
0. 14
0.04
0. 18
0. 18*
0.06
0.06
NSC
0.07*
<0. 04
0. 13
0.23
NSC
NSC
0.06
<0.04
NSC
NSC
0. 52
<0.04
NSC
Period 5
NSC
NSC
NSC
NSC
< 0.04
NSC
0. 08
< 0.04
0.31
0. 10
< 0. 04
0. 12
< 0.04
0. 07
0. 12
< 0.04
NSC
0. 36
IS
0.31
< 0.04
NSC
0. 10
NSC
NSC
NSC
NSC
NSC
< 0.04
0.06
< 0.04
0. 16
0.23
NSC
0.08
0. 13
NSC
NSC
NSC
NSC
NSC
NSC
<0. 04
< 0. 04
< 0. 04
NSC
0. 10
0. 25
<0. 04
0.05
Zinc
Period 1
126*
NSC
NSC
196*
142
NSC
151
89
80
81
96
108
70
' 59
81
63
153
151
61
96
113
97
102
NSC
46
77
124
149
113
144*
97
96
97*
125*
85
120
NSC
39*
66*
68
85
NSC
NSC
108
110
NSC
NSC
174*
99
NSC
-------
TABLE A-21 (continued)
Participant Cadmium
I.D. Period 5
600 0.157
601 0. 165
60Z 0. 066
603 0. 169
614 0.033
615 0.071
619 0.029
621 0.205
622 0. 048
623 0. 057
628 0.034
630 • 0.054
631 0.113
632 '0.211
634 ' 0.287
640 - 0.016
641 0. 068
642 0. 127
643 0. 143
644 0.014
645 0.125
646 0. 159
647 0.067
648 0. 048
650 0. 199
651 0. 105
653 0.027
654 0. 105
-------
TABLE A-22 TRACE METAL CONCENTRATIONS IN HAIR SAMPLES
FROM INDIVIDUAL PARTICIPANTS Og/g)
Participant Cadmium Copper Lead Mercury Zinc
I. D. Period 1 Period 5 Period 1 Period 1 Period 5 Period 1 Period 5 Period 1
1 0.68 0.91 26.4 12. 1 13.2 0.67 0. 8Z 307
2 <0. 13 <0. 13 12.0 4.6 1.7 0.42 0.27 232
3 0.41 0.31 15.5 12.0 11.4 0.45 IS 155
4 1.48 1.06 18.1 24.4 Z9. 1 0.72 IS 108
5 0.48 0.53 NSC NSC 17.4 0.67 IS 123
6 0.47 1.43 34.9 5.6 17.6 0.36 0.64 226
9 0.93 0.25 36.7 12.5 6.8 0.62 IS 110
10 0.63 0.52 25.0 12.7 7.4 0.24 IS 162
11 0.43 <0. 13 14.2 4.8 9.4 1.34 IS 143
12 0.26 <0. 13 12.7 13.6 9.5 0.50 , IS 342
13 2.10 0.25 56.8 24.9 48.7 0.19 IS 770
14 0.18 0.43 12.6 7.1 4.8 0.46 IS 221
15 0.27 0.13 12.0 2.9 5.0 0.70 IS 192
16 0.48 1.83 15.5 15.8 75.8 1.37 IS 273
18 0.81 1.03 21.4 24.7 29-1 0.73 0.33 91
19 0.38 0.17 19.2 6.6 6.5 0.54 IS 127
21 NSC NSC NSC NSC NSC NSC NSC NSC
29 0.58 0.32 67.8 5.9 6.2 0.49 IS 162
30 0.42 0.59 31.6 7.9 <0. 9 0.38 0.21 112
31 0.32 0.51 35.0 4.0 4.3 0.48 <0. 17 162
32 0.83 0.36 22.9 11.2 7.7 0.50 IS 98
33 0.95 1.03 21.0 14.4 16.1 0.37 0.27 45
49 0.15 NSC 149.3 5.2 NSC 0.65 NSC 223
50 0.68 2.18 56.4 10.1 7.8 0.38 IS 188
70 NSC NSC NSC NSC NSC . NSC NSC NSC
74 1.21 0.33 15.2 11.4 11.3 0.60 IS 116
75 <0. 13 <0. 13 93.7 2.2 3.4 0.95 0.59 186
76 1.20 0.96 24.3 39.0 19.3 0.84 0.34 189
77 0.55 0.15 22.4 23.8 5.1 0.47 IS 68
78 2.20 3.14 20.6 11.7 26.6 1.79 0.42 102
83 <0. 13 0.14 52.7 2.9 1-0 <0. 17 <0. 17 38
84 <0. 13 <0. 13 28.1 3.6 4.6 0.19 0.35 169
97 1.41 0.43 51.1 20.6 18.7 0.41 0.31 176
98 1.37 1.71 19-5 16.6 22.0 0.55 IS 85
99 1.39 1.14 60.0 26.2 24.5 1.12 0.47 148
100 1.25 0.20 43.1 8.9 5.2 1.69 IS 150
101 0.60 0.72 27.4 37.8 358.8 0.52 0.58 102
102 0.43 0.15 22.1 4.6 2.8 1.22 0.90 183
103 1.20 0.46 63.4 11.8 5.3 0.35 0.47 128
113 0.55 NSC 52.7 5.4 NSC 0.47 NSC 165
114 0.63 NSC 96.9 9.5 NSC 0.55 NSC 125
115 0.36 NSC 52.4 5.0 NSC 0.22 NSC 126
116 0.83 NSC 71.6 7.7 NSC 0.27 NSC 140
117 3.94 ' NSC 71.6 4.0 NSC 0.81 NSC 144
131 1.60 0.29 72.0 12.6 3.4 0.64 0.99 150
(continued)
-------
TABLE A-22 (continued)
Participant
I. D.
141
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
(continued)
Cadmium
Period 1
1.42
1.24
0. 93
0. 64
<0. 13
<0. 13
1. 14
<0. 13
0.78
1. 19
<0. 13
0. 57
0. 16
0. 18
1.46
1.41
1.45
1.51
3.53
3.29
0.71
<0. 13
0.92
NSC
0. 14
0. 62
0.78
0.41
4.05
1.91
1.76
1.01
0.37
<0. 13
1.27
1.08
0.26
2.28
0.88
<0. 13
0.38
3.90
0.26
0. 13
0. 64
0.48
Period 5
0.74
0. 53
0.84
0.24
0. 59
0. 13
0. 34
<0. 13
0.83
0.41
0. 30
0.24
0. 13
NSC
1. 31
1.28
0.88
0.20
2.20
0.86
0.43
<0. 13
0. 60
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0. 59
0.88
0. 18
NSC
NSC
0. 17
0.29
0.42
0.27
0. 18
14.39
<0. 13
0. 19
0.26
0.23
1. 10
Lead
38.4
50.0
33.2
48.7
185.7
97.7
50. 1
28. 9
17.3
24.3
128.2
25.2
19.2
32.3
59. 9
32. 5
60. 6
196.7
182.2
74.4
44.0
31.3
23.7
NSC
113.6
14.4
39. 1
15.7
18.8
20.3
35.2
29.5
57.0
44.2
26.6
119.4
88.2
84.7
55. 1
.66.5
39.0
37.2
49. 6
71. 5
55.2
66.2
Period 1
24.7
31. 1
17.9
6.6
9.0
4.7
14. 5
8.0
9-7
18. 2
4. 7
11.1
0. 9
9.9
29. 1
10. 5
17. 0
6.2
31. 6
19.2
18. 6
<0. 9
17.4
NSC
2.0
14.9
15.2
17. 6
10. 9
83.2
18. 1
27.9
7.7
1.0
15.9
8.9
4. 3
39.7
12.0
<0. 9
9.8
4. 5
3. 9
4. 6
31. 3
11. 6
Period 5
12.2
13. 5
8.0
6.2
7. 5
1.9
9.5
5.8
20.4
20.9
3.8
5.9
3.7
NSC
30.0
8. 7
18. 5
3.0
10. 5
15.3
11.8
1. 7
13. 7
NSC
NSC
NSC
NSC
NSC
NSC
NSC
6.0
12.0
5.2
NSC
NSC
9.9
6.4
11. 3
4. 3
3. 9
2.3
2. 1
4. 6
5.8
9. 3
11. 3
Mercury
Period 1
1.40
1. 14
1. 19
1. 17
1.47
0.72
0. 59
0.49
0. 47
1. 21
0.37
1. 31
0. 69
0.40
1. 11
0.49
0. 51
0. 81
1. 31
0. 32
0. 35
0.24
0.32
NSC
0.28
0.29
0.28
0.70
0. 19
0. 27
1.09
1. 13
1.69
0.76
1.52
0. 59
0. 65
0.35
0. 40
0.71
0.34
<0. 17
0. 17
0. 24
<0. 17
<0. 17
Period 5
IS
0. 69
0. 68
IS
IS
0.46
IS
IS
IS
IS
IS
0. 17
0. 17
NSC
IS
IS
IS
0. 17
0.48
IS
0.26
IS
IS
NSC
NSC
NSC
NSC
NSC
NSC
NSC
0.49
0. 54
IS
NSC
NSC
0. 32
0. 55
0.43
0.21
IS
0.21
IS
IS
IS
0. 33
1. 10
Zinc
Period 1
141
336
141
147
365
126
140
147
73
71
171
139
172
122
70
145
128
191
163
62
139
165
43
NSC
169
53
57
195
2, 132
960
93
166
109
196
481
177
160
125
130
188
99
2,006
122
186
133
93
-------
TABLE A-22 (continued)
Participant
I. D.
231
232
233
235
239
241
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
295
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
(continued)
Cadmium
Period 1 Period 5
<0. 13
0.66
4. 69
2.07
NSC
1.41
0. 53
0. 36
0. 65
NSC
1.96
0.36
1.24
<0. 13
1.44
0.36
0.70
1.05
0.23
0.79
2.21
1.86
2.44
3.49
1.45
0.46
0. 16
0.32
0.86
0.46
1.77
1. 19
2.94
0. 13
1. 13
<0. 13
2.33
<0. 13
1. 12
0.82
0.43
1.75
0.73
1.43
0.32
0. 16
Z. 11
<0. 13
1.39
NSC
<0. 13
0.23
0.21
1.86
NSC
NSC
NSC
NSC
NSC
0. 68
0. 15
0. 51
0.51
0.29
0.20
0. 51
0. 69
0.35
3.07
1.88
0.35
NSC
0.35
NSC
0.70
0.31
5.31
2.75
0.76
0.46
0. 18
<0. 13
0.25
0.55
1.03
0. 15
0.2
0.44
1.94
0.27
Lead
71.2
17.9
71.2
46.3
NSC
158.2
36.6
22.3
28.2
•NSC
108.2
161. 1
52.3
94.5
34.4
20.4
16.3
15. 5
33.7
47.2
25. 6
34.6
37.9
39.8
20. 6
11.9
71.2
14.0
13. 6
29.3
13.7
19.0
37.6
17. 1
11.6
11.9
16.5
10. 1
18.7
41.2
60.0
75.4
26.9
22. 5
123.9
Period 1
1.7
13.0
3.3
22.4
NSC
8. 1
3.2
8.4
17.8
NSC
31.2
4. 6
18.0
47.0
26.7
164.6
223.3
4. 6
6.6
6. 1
306.0
9.7
13.3
21.9
17. 1
12. 1
6.3
9^ 6
8. 1
2.6
2.2
29.0
32.4
<0.9
13.4
3.7
2. 6
4.7
5.5
25.4
11.8
10.0
6.8
12.7
13. 1
Period 5
1.9
21.4
4. 1
21.3
NSC
4.3
6.3
11.3
29.0
NSC
NSC
NSC
NSC
NSC
56.6
68.3
13. 1
7.0
2.0
5. 1
3.0
5.7
3.7
14.7
20.0
11.0
NSC
5.8
NSC
18.7
9.0
91.3
98.0
8. 1
6.5
4.3
1.8
5.2
3.9
48.7
3.5
1.6
3.9
16.3
24.2
Mercury
Period 1
0.27
0. 19
<0. 17
0. 45
NSC
0. 61
1.04
0.29
0.22
NSC
1.46
0.21
1.46
0.44
0.29
0. 56
0.46
0.31
0. 58
0.41
0.45
0.77
0. 56
0.33
0.36
0.72
0.93
0. 45
0.20
0.21
<0. 17
0. 90
0.38
0. 55
0. 55
0.43
0. 59
0. 55
0.24
0.43
1.03
36.05
1.87
2.66
2. 15
Period 5
2. 50
IS
<0. 17
IS
NSC
0. 58
IS
IS
IS
NSC
NSC
NSC
NSC
NSC
0. 19
0.86
0.21
IS
0.84
0.21
IS
0. 63
0.22
<0. 17
IS
0. 64
NSC
IS
NSC
0. 20
IS
0.41
IS
0.30
IS
IS
IS
IS
IS
0.22
0. 38
0.26
IS
0.80
0. 29
189
112
,408
135
NSC
119
157
123
134
NSC
117
272
124
165
114
163
293
163
167
161
57
208
225
159
116
137
176
, 546
171
151
,031
165
534
141
390
396
109
191
151
160
144
129
142
101
109
-------
TABLE A-22 (continued)
Participant
I. D.
340
341
342
343
348
349
350
353
354
355
356
358
359
361
362
363
364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
(continued)
Cadmium
Period 1
0.79
0. 68
1.41
1.28
<0. 13
<0. 13
NSC
1.54
1.63
1. 68
0. 94
1.00
5.83
<0. 13
0.49
<0. 13
0.45
9.59
1. 14
1. 60
<0. 13
0.71
0.26
2.30
2.30
0.63
0.32
0.46
5. 10
0.33
<0. 13
0. 13
0.65
1.81
0.24
0.60
1.57
0.47
3.46
0.77
0.87
0.35
0.20
0.26
0.45
Period 5
0.41
0.87
0. 16
0. 89
0. 62
<0. 13
<0. 13
0. 17
0.85
0.81
0.37
NSC
<0. 13
0.23
0.49
0. 57
C.29
4.48
6.23
0.33
1. 15
0.75
0. 14
0.42
1.56
1.23
0. 50
NSC
NSC
0.39
0.29
<0. 13
1.74
1. 15
. 1. 13
0.29
0.47
NSC
NSC
NSC
NSC
0. 31
<0. 13
0.22
NSC
Lead
11.8
14. 6
28.4
21.3
13.0
10. 5
NSC
28. 1
17. 9
13. 9
14.8
16.4
27.8
75.2
15.2
21.3
34.0
20.2
20. 6
40.7
13.6
11.8
14.0
96.5
37.0
43.8
11.4
16. 6
10. 5
78.3
19.6
38.4
12.9
28.6
16.4
82.2
27. 1
14.2
34. 6
19-2
33.2
42.4
30.3
19.4
99.9
Period 1
16. 1
20. 2
42. 2
71. 3
6.8
3. 1
NSC
3.4
17. 7
14. 9
20.8
1. 4
398.0
2. 9
14. 1
8. 2
11.9
24. 0
3. 0
22. 1
5.7
18. 6
4. 0
9.8
12. 1
14. 7
12. 0
8. 6
9.6
4. 5
7. 3
10. 5
8. 7
11.8
16. 5
3. 6
8. 8
5. 5
13. 4
14. 9
8. 3
18. 6
<0. 9
2.4
5. 5
Period 5
7.4
9.3
<0.9
21.4
21. 5
<0. 9
<0.9
<0.9
16.8
12.4
11.8
NSC
1.7
2.9
<0. 9
11.2
4.3
27.9
13.7
4.3
13.0
5.9
1.6
3.3
20.0
6.0
11.2
NSC
NSC
3.5
15. 5
2.3
14. 3
2.7
12. 1
5.9
6.7
NSC
NSC
NSC
NSC
9.8
<0. 9
5.7
NSC
Mercury
Period 1
1.20
1. 12
0. 28
0. 52
0. 59
0.25
NSC
0. 57
0.73
0. 59
0.44
0. 34
0. 60
1.91
1. 32
0. 54
1.22
0. 37
0.35
0. 50
0. 18
0. 36
0.22
0.33
0. 68
0.49
0. 54
0. 96
0.24
0.25
0. 65
0.32
0.26
0. 36
0. 29
0. 17
<0. 17
0. 34
0. 34
0. 36
0. 39
1. 34
0.81
0.37
0.24
Period 5
0.25
0. 39
IS
0. 72
0.22
IS
0.40
<0. 17
IS
0.84
0.39
NSC
0.75
IS
IS
IS
<0. 17
0.48
0.22
IS
0. 32
IS
0.47
0.27
IS
0. 94
IS
NSC
NSC
IS
IS
IS
IS
0. 52
0.28
IS
0. 22
NSC
NSC
NSC
NSC
0. 50
1. 60
0. 30
NSC
Zinc
Period 1
45
42
989
115
296
123
NSC
651
76
96
141
670
2,591
170
66
65
149
4,306
264
669
180
114
143
153
873
142
Z33
130
2,939
191
115
159
177
595
6,339
237
185
229
269
72
472
221
199
163
191
-------
TABLE A-22 (continued)
Participant
I. D.
403
405
406
407
408
41Z
413
414
415
416
417
418
419
4Z1
422
423
424
426
428
OJ 429
0 43°
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
(continued)
Cadmium
Period 1
0.26
<0. 13
<0. 13
<0. 13
0.46
0.52
0.36
0.86
0.22
0.86
<0. 13
3.58
0.26
0.68
0.48
<0. 13
1.66
0.40
0.36
0. 62
3. 16
<0. 13
1.26
2. 67
3. 14
2.49
3.77
2. 38
1.47
9-94
1.43
0.39
0.23
2.33
1. 14
2.79
1.00
0. 18
0.21
0.41
3.09
0.35
<0. 13
1.06
Period 5
0. 18
1. 76
0. 31
0. 40
0. 37
NSC
NSC
0.86
0.26
0.47
<0. 13
3. 13
0.23
0.84
2. 52
0. 50
2.91
2. 04
0. 31
<0. 13
0.89
NSC
1.45
IS
1. 51
1. 18
1. 37
2. 51
1. 60
1. 55
NSC
NSC
NSC
1. 19
<0. 13
0.48
0. 50
0. 54
<0. 13
<0. 13
<0. 13
NSC
NSC
0. 50
Lead
32.8
14.8
24.0
22. 5
20. 5
28.8
95.0
16.7
56.6
16.0
24. 1
21.3
48.5
27. 1
25.7
25.6
11.3
31.2
28. 2
13.6
13.4
30.2
24.8
18.8
14.8
16.0
17.3
49. 6
80. 5
120.0
60.6
23. 1
30.8
14.6
33.7
22.9
27.6
16. 6
21.2
14.6
33. 1
11.9
23.9
14.9
Period 1
1. 3
11. 9
2.4
1.0
15. 9
9.0
21.6
15.2
7. 1
16. 6
8.4
67.8
5.4
13. 1
2.4
5.4
27.0
5.2
6.5
13.3
20. 6
2. 9
20.3
23.8
39.2
27. 6
43. 9
19. 3
12.6
241. 6
4. 1
10.7
9.2
30. 6
24.4
25. 1
11.8
1. 6
2.2
6.4
5.8
20.0
5.2
23. 5
Period 5
6.9
8. 5
<0. 9
3.3
17.9
NSC
NSC
18. 5
10.8
10.4
2.9
50. 6
2.4
8. 5
17. 5
<0. 9
19. 1
1.3
4. 5
2. 9
13. 1
NSC
5.8
IS
14.9
6.0
42.8
42.2
18.7
18.4
NSC
NSC
NSC
12.7
3.7
10.2
14.3
2.4
<0. 9
12.9
2.0
NSC
NSC
19. 5
Mercury
Period 1
0.35
0. 62
0.22
0. 18
0. 35
<0. 17
0.21
0. 67
0. 34
0. 52
0.48
0. 28
0.46
0. 39
0.81
0. 46
1.30
1. 13
0. 56
0. 44
0. 20
0. 90
2.27
2.48
1.07
1. 33
1.47
0. 26
0.89
IS
0. 54
0.47
0. 67
0.47
0. 50
0.37
0.23
<0. 17
1. 19
0. 20
0.26
0.77
0. 50
0.83
Period 5
IS
0. 47
IS
<0. 17
IS
NSC
NSC
0. 88
0. 48
0. 66
IS
0. 91
0. 30
0. 92
IS
IS
IS
<0. 17
0. 17
<0. 17
<0. 17
NSC
IS
IS
IS
0. 25
0. 38
IS
IS
IS
NSC
NSC
NSC
IS
IS
IS
<0. 17
0.33
IS
0.48
0.21
NSC
NSC
0. 85
Zinc
Period 1
130
142
187
174
127
228
188
92
233
90
179
653
176
50
184
132
65
129
152
132
99
153
394
187
167
266
275
215
582
106
289
138
216
114
143
186
97
160
118
142
383
132
150
104
-------
TABLE A-22 (continued)
Participant
I. D.
501
502
503
504
505
506
507
508
509
510
512
Cadmium
Period 1
0.43
0.74
1.00
0. 17
1. 15
0.91
0.23
1. 65
0.42
3.32
NSC
Period 5
NSC
NSC
NSC
0. 16
0. 61
3.01
0.27
1.30
0. 15
0.47
0. 68
Lead
7.9
105.7
85.8
14.9
30.8
24.2
17.9
31. 6
55. 1
12. 6
NSC
Period 1
9.4
9.9
8.6
2. 1
7.9
20.6
13.8
18.2
5.8
29- 1
NSC
Period 5
NSC
NSC
NSC
3.2
8.3
35.8
9.8
18.4-
3.7
5.9
6.0
Mercury
Period 1
0.41
0.39
0.62
<0. 17
1.06
1.87
0.80
1.81
0.76
0. 31
NSC
Period 5
NSC
NSC
NSC
IS
0.45
0. 29
IS
1.40
IS
<0. 17
0. 56
Zinc
Period 1
56
203
220
143
39
40
79
172
112
122
NSC
-------
TABLE A-23
Participant
I. D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
21
29
30
31
32
33
49
50
70
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
(continued)
Cadmium
Period
1. 00
O.S1
0.74
0. 86
0. 84
0. 98
0.62
1. 05
0. 18
0. 70
0. 91
0. 92
0. 24
0.72
0.41
0. 56
0. 18
1. 10
0.33
0. 59
1. 05
0. 55
0.61
0. 57
NSC
0. 59
0.26
0.32
0.72
NSC
1. 19
0. 79
0.85
0. 15
1. 18
0.39
0.63
0.75
1. 23
NSC
0. 28
1. 26
0.47
0. 15
0.38
0. 44
0. 59
1 Period 5
0.32
0.72
. 0.44
1. 53
0. 18
0.24
0.36
1. 04
0. 90
0.42
0. 60
0.61
0.33
1.40
0.22
0.63
NSC
0.79
0.37
0. 17
0.42
< 0. 13
0.02
0. 57
NSC
1.15
0.60
0. 55
0. 97
0. 24
< 0. 13
0. 51
0.71
0. 23
0. 25
0.28
1. 14
0.22
0.88
NSC
NSC
NSC
NSC
NSC
0.39
0.38
0.83
TRACE METAL CONCENTRATIONS IN URINE SAMPLES
FROM INDIVIDUAL PARTICIPANTS (jug/1)
Copper Lead
11.2
12.4
15.0
10. 1
9.9
10. 1
7. 1
18.0
5. 4
7.4
9.7
8.4
3. 2
9.8
11.3
12.4
9.2
4.6
11.7
11.4
6.7
15.4
9.2
10.3
NSC
9.6
13.7
9.2
14. 5
NSC
12.6
9.0
11.2
8.3
11.0
2.4
9.2
9.5
14.9
NSC
3.7
10.4
16.2
4.3
8. 1
15.2
17. 2
Period 1
10.5
14.0
40.3
7.7
6.7
14. 2
8.8
8.6
3.7
10. 5
12. 1
11.6
3. 5
11.7
10.5
5.7
10.3
6. 1
14.0
6.8
23.9
14.9
9.2
10. 3
NSC
9.6
13. 7
9.2
14. 5
NSC
12.6
9:0
11.2
8.3
11.0
2.4
9.2
9.5
14. 9
NSC
3.7
10. 4
16.2
4.3
8. 1
6. 5
9.8
Period 5
<2. 1
10. 1
4.5
4.9
<2. 1
2.4
9. 1
7. 1
4.4
4. 0
5.6
9. 1
2. 4
6. 5
8.2
5.6
NSC
5. 1
4.8
4.2
<2. 1
3.2
8.8
5.3
NSC
11.3
3. 1
5.6
5. 0
4. 4
<2. 1
<2. 1
4.2
6.8
<2.09
8. 1
27.7
3.0
10.6
NSC
NSC
NSC
NSC
NSC
3.3
5. 3
<2. 1
Mercury
Period 1
8.29
9. 33
10.37
8.29
9.85
5.47
8.81
16. 59
9.85
9.85
9.85
8.81
7. 78
7.26
10.89
10.37
10.37
8.81
10.37
7.26
7.78
8.81
9. 85
4.77
NSC
8.29
4. 15
8.24
10.89
NSC
7.78
8.81
8.81
8. 29
7.26
1. 17
2.97
12.85
12.85
NSC
2.97
1.89
1.35
3. 14
5.02
8. 54
19. 07
Period 5
< 1. 14
2.88
1.81
< 1. 14
< 1. 14
2.63
3. 54
3.37
1.81
< 1. 14
<1. 14
2.24
<1. 14
2.89
4.64
5. 24
NSC
5. 08
1.91
1.67
2. 21
1.47
1.83
4. 10
NSC
1.38
< 1. 14
<1. 14
< 1. 14
< 1 . 14
< 1. 14
<1. 14
<1. 14
1. 30
< 1. 14
2.66
<1. 14
5.46
<1. 14
NSC
NSC
NSC
NSC
NSC
2. 15
2. 24
1. 89
Zinc
Period 1
365
410
265
287
391
954
205
520
137
397
442
128
301
556
456
237
465
132
328
511
465
451
267
1000
NSC
322
207
437
354
NSC
598
529
203
202
460
483
497
675
560
NSC
138
364
331
189
331
595
489
-------
TABLE A-23 (continued)
Participant
I. D.
143
144
145
146
157
158
159
160
162
165
166
167
171
172
177
178
179
180
181
182
183
189
190
191
192
193
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
(continued)
Cadmium
Period 1
0.39
0. 28
0.35
0.61
0. 73
0.62
0. 20
0.89
0.60
0. 9.8
0.33
0.30
0. 48
0.45
1.46
0.83
0.71
0.36
2.48
1.38
0. 50
0.61
0. 14
0. 21
0. 15
0. 18
0. 57
0. 57
0.48
0. 44
0. 27
0.28
< 0. 13
0.48
0. 51
0. 19
0.63
0.73
0.72
0. 42
1.07
0. 90
0.39
0. 49
0.28
< 0. 13
0.62
Period 5
0. 50
0. 58
0. 14
0. 91
2. 06
0. 27
0.73
0.51
0. 50
0. 50
1. 10
NSC
1. 16
0.23
1. 15
0.78
0.37
0.34
1.63
0.63
0.66
NSC
NSC
NSC
NSC
NSC
NSC
NSC
1. 03
0. 42
0. 82
NSC
NSC
0. 47
0. 72
0.38
1.31
0. 76
0. 96
1.45
1.39
0. 97
0.80
1.42
0. 20
1. 21
0. 23
Lead
8.6
16.8
2.9
7.8
8.9
18. 0
7. 0
43.8
11. 5
13.0
13.8
10.8
7. 1
5. 5
7. 1
2. 7
10.4
15.4
20.8
16. 8
6.4
14. 1
5. 2
5.4
10. 5
10. 8
18. 0
17.0
8. 7
12. 7
7.9
15.9
15. 2
13.8
14.9
12.-5
11.6
10. 5
5. 5
13.9
11.7
12. 1
11.5
14. 2
11.1
17. 2
6. 1
Period 1
10. 5
6.8
3. 5
2. 5
20. 8
9. 5
4. 8
6.8
4. 0
4. 0
12.9
<2. 1
8.7
7. 7
5.9
<2.1
14. 1
11.8
22.6
12. 5
5.0
18. 7
3.6
4. 8
8.7
8.7
13. 1
8.7
7. 1
10. 1
2.3
2.6
11.0
7. 5
17.7
11.9
17. 0
11.0
12. 5
11.0
23.3
15.2
10. 1
6.6
8.3
8. 4
2. 5
Period 5
2.4
4.6
<2. 1
3. 1
13.3
3.7
<2. 1
4.9
3.4
6. 1
6. 1
NSC
3. 5
9.2
3.6
4.2
<2. 1
8.4
15.6
6.8
11. 5
NSC
NSC
NSC
NSC
NSC
NSC
NSC
7.3
9.5
4.0
NSC
NSC
<2. 1
17. 5
<2. 1
10.2
5. 5
4.6
12. 1
18.8
7. 5
12.9
9. 9
<2. 1
4.2
2. 5
Mercury
Period 1
3.09
2.39
2.07
8.3Z
< 1. 14
2.67
1. 55
3.09
< 1. 14
6.36
2.93
4.64
< 1. 14
2.93
< 1. 14
< 1. 14
2.39
1.96
2.39
10.27
.9.78
4.35
5.38
5.87
3.09
6.85
< 1. 14
2.39
8.32
10.27
9.78
8.32
9.29
10. 89
10.37
3.37
3.08
12. 44
6.85
12.23
7.83
5.41
4.44
2. 12
2. 50
<1. 14
3.67
Period 5
< 1. 14
1.91
1. 72
1.30
1.89
< 1. 14
< 1. 14
< 1. 14
2. 32
7. 20
1.91
NSC
< 1. 14
2.68
1.26
<1. 14
1.21
3. 13
1.24
6.41
1.64
NSC
NSC
NSC
NSC
NSC
NSC
NSC
6.41
1.67
1.59
NSC
NSC
< 1. 14
2.64
< 1 . 1 4
<1. 14
<1. 14
<1. 14
8.54
1. 59
4. 02
2.32
2. 15
2. 58
1. 59
1.83
Zinc
Period 1
427
812
450
115
473
950
312
7406
441
473
363
393
560
386
175
142
312
268
672
622
346
986
134
489
521
504
844
986
346
480
286
253
515
281
557
871
543
379
271
290
489
337
529
454
337
206
84
-------
TABLE A-23 (continued)
Participant
I. D. .
235
239
241
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
295
302
303
312
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
344
(continued)
Cadmium
Period 1
1. 17
NSC
0.46
0.33
2. 51
0.33.
0.33
< 0. 13
0.37
0.35.
1. 00
0.36
1. 11
0. 52
0:35
0.82
0.77
1. 88
0.28
0.45
0.78
0.42
0.38
1. 40
1.38
NSC
0.40
1.26
0.63
0.81
4. 12
0.64
0. 58
1.67
0. 24
0.98
NSC
0.87
0. 57
0. 19
0.36
0.71
0. 71
0.41
0.69
0.60
1. 26
Period 5
0.31
NSC
0.48
1.37
0. 80
0.98
NSC
NSC
NSC
NSC
NSC
0.91
1. 15
0.55
1.37
0. 17
< 0. 13
0.98
<0. 13
0.22
1. 00
1.29
0.39
NSC
1.85
NSC
0.30
1.24
0. 55
0. 28
0.86
0.44
0.94
1. 15
0.88
0.98
0.44
0. 18
0.35
0. 22
<0. 13
0. 95
0. 97
1. 22
0.49
0.38
NSC
Lead
16. 5
NSC
16.4
13.8
7.4
29.7
10.2
4.3
11.4
4. 5
5.0
12. 1
16.2
6.7
6.4
11.7
18.9
19.5
5.5
5. 5
11.9
2.8
8.4
14.6
11.4
NSC
5. 1
4. 5
7. 1
12.9
19.1
16.8
7.5
9.5
5.7
8.2
NSC
16.9
15.4
10. 1
14. 1
6.7
7.8
18. 5
8.4
7.8
15.4
Period 1
16.7
NSC
10.6
14. 1
4. 2
16.3
11.9
19.7
9.4
8.4
6.0
10.6
30.8
9.4
6.0
15.2
10. 1
17.4
7.5
7.5
6.6
7.5
13.3
28.2
4.8
NSC
4.3
8. 5
22.4
10.4
6.7
14.9
2.4
9.2
4.2
4.8
NSC
7.6
9.2
10. 1
14. 1
6.7
7.8
18.5
8.4
7.8
15.4
Period 5
3.0
NSC
?.9
3. 1'
3.8
7.3
NSC
NSC
NSC
NSC
NSC
7.6
12.1
11.9
8.0
5.6
5;3
9.6
<2.1
2.4
7. 2
10. 9
3. 1
NSC
6.5
NSC
7.8
8.4
8.4
2.9
5.0
8.5
3. 5
8.0
4.6
13. J
10.7
14. 1
8.0
8.0
3.4
2.1
8.0
7. 1
<2. 1
4.3
NSC
Mercury
Period 1
2.89
NSC
3.86
12.72
4.63
4.82
6.56
2. 12
2.70
1.93
3.09
3.09
13.74
2.51
<1. 14
21.99
10.74
2.67
1.54
1.68
5.38
1.54
4.24
7.72
3.28
NSC
3.28
2.70
3.28
3.09
2.51
<1. 14
1.93
3.90
4.44
2.70
NSC
6.45
3.67
2.70
7.91
2.83
3.09
2.70
2.51
5.31
2.39
Period 5
<1.14
NSC
<1. 14
3.47
<1. 14
<1. 14
NSC
NSC
NSC
NSC
NSC
<1. 14
2.81
1.24
1. 55
1.75
<1. 14
<1. 14
<1. 14
<1.14
<1. 14
1.79
2.07
NSC
<1. 14
NSC
2. 15
1.26
2.00
3.05
<1. 14
1.30
<1. 14
< 1 . 1 4
2. 15
2.40
1. 21
<1. 14
1.30
1.24
1.55
<1. 14
<1. 14
1.81 .
1.55
<1. 14
NSC
Zinc
Period 1
477
NSC
285
529
134
471
424
375
564
328
92
880
844
342
308
755
1412
1291
360
456
447
190
377
638
295
NSC
524
181
421
746
481
614
143
557
562
114
NSC
1351
561
538
561
357
482
491
419
3?7
982
-------
TABLE A-23 (continued)
Participant
I. D.
348
349
350
353
354
355
356
359
361
362
363
364
365
'366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
407
408
412
413
(continued)
Cadmium
Period 1
1.20
0. 82
0.35
0. 72
0. 90
0.25
5. 05
1. 72
0.70
0. 59
0.89
2.43
0.26
1. 05
0.69
0.48
0. 83
0. 76
0. 50
0.80
0.60
2.42
1.07
1.62
0.61
0.50
0.47
0. 97
0.85
0. 97
0.31
0.64
0.60
0. 52
0.58
0. 80
0. 18
0. 90
0. 22
1. 13
2. 28
0.22
0. 56
1. 51
1.83
0.76
0. 56
Period 5
1.41
0. 77
0.67
0. 15
< 0. 13
0.33
0. 23
o;66
0. 49
< 0. 13
0. 20
0. 17
0. 15
1.32
0. 77
0. 47
1.02
0.86
< 0. 13
1.03
< 0. 13
1.07
NSC
NSC
0.38
0.34
1. 52
0.82
0. 19
0.76
0.78
0.88
NSC
NSC
NSC
NSC
0. 74
0.83
0. 57
NSC
0. 74
0. 26
0.30
0.85
0.37
NSC
NSC
Lead
21.7
19.6
14.6
5. 1
10. 2
9. 7
24.4
12.2
13.6
15.7
13.6
13. 8
7.8
12.3
7.8
12.9
8. 1
13.2
5. 5
11.5
7. 0
8.9
8.9
14.6
14. 5
7.0
7. 5
11.5
15.2
13. 1
9.4
14.2
15.2
<2. 02
20.9
10.6
5. 1
14.7
4.9
14.9
10.6
7. 2
12. 5
18.9
19.3
8.2
6.6
Period 1
21.
19.'
14.
5.
10.
9.
24.
12.
13.
15.
13.
13.
7.
12.
7.
12.
8.
13.
5.
11.
7.
8.
8.
14.
14.
7.
7.
11.
15.
13.
9.
14.
15.
<2.
20.
10.
6.
15.
6.
20.
15.
11.
10.
10.
27.
5.
10.
7
6
6
1
2
7
4
2
6
7
6
8
8
3
8
9
1
2
5
5
0
9
9
6
5
0
5
5
2
1
4
2
2
02
9
6
8
6
2
1
6
Z
5
9
9
5
5
Period 5
4.5
10. 1
5.3
<2. 1
<2. 1
5.4
8.0
3.3
9.0
2.4
3.9
<2. 1
2.6
8.4
7.6
6.0
5.2
14.3
3.3
5. 1
3. 9
6.7
NSC
NSC
13. 7
7.9
3. 5
7.6
6.0
22. 1
2.8
5.0
NSC
NSC
NSC
NSC
6.8
5.5
8. 1
NSC
11.6
4. 2
<2. 1
4.6
3.7
NSC
NSC
Mercury
Period 1
4.
8.
3.
2.
3.
5.
5.
1.
3.
1.
2.
4.
6.
7.
6.
5.
10.
1.
8.
7.
10.
1.
1.
2.
2.
2.
<1.
< 1.
2.
3.
Z.
2.
1.
1.
1.
6.
<1.
6.
16.
8.
<1.
6.
11.
2.
9.
2.
<1.
05
68
09
51
67
89
4
54
09
42
70
24'
56
79
17
40
61
42
15
72
27
54
35
89
70
27
14-
14
83
47
51
81
26
54
54
87
14
02
24
15
14
73
08
48
21
27
14
Period 5
1. 89
6.62
2.21
1.65
1.34
2.31
1.89
<1. 14
2.78
<1. 14
< 1. 14
<1. 14
< 1 . 1 4
1.38
1.91
<1. 14
1.65
1.21
<1. 14
1.37
1.55
<1. 14
NSC
NSC
2.58
2.63
<1. 14
<1. 14
3.26
<1. 14
1.47
< 1. 14
NSC
NSC
NSC
NSC
4.20
4.22
5.25
NSC
3.89
2.75
3. 19
1.91
3.26
NSC
NSC
Zinc
Period 1
798
620
509
536
285
912
719
256
329
439
536
667
430
541
570
441
329
461
560
538
306
264
774
817
589
403
422
435
468
454
556
380
422
460
669
491
269
482
314
694
420
565
455
678
950
212
159
-------
TABLE A-23 (continued)
OS
Participant
I. D.
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
512
Cadmium
Period 1
1. 07
1.43
0.43
0.66
0.31
1. 22
0.81
1.99
0.69
1. 17
2.60
0.38
1. 11
0.95
1. 44
1.05
0.45
0.63
0.37
NSC
0. 59
1. 17
NSC
0. 18
1.06
0.77
1. 00
0. 95
0.47
1.08
0.47
1.35
1. 17
0. 72
0.30
0.88
NSC
0. 56
0. 56
0. 18
0.87
0. 17
0. 51
0.90
NSC
0.39
0.73
NSC
Period 5
0. 49
0. 16
0.66
0. 77
0. 84
0. 85
0. 77
0.62
1. 91
0. 20
0. 70
0. 43
0.60
0. 14
NSC
0. 38
0. 18
0. 19
0. 28
0.79
0. 81
0. 47
NSC
NSC
NSC
NSC
0. 87
0. 76
0. 14
0. 22
0. 14
0. 21
0. 96
0. 17
NSC
NSC
1.82
NSC
NSC
NSC
0. 84
0. 21
0. 47
<0. 13
0. 36
NSC
0. 55
0. 28
Lead
9.9
13. 1
10.3
4.9
7. 2
13. 1
6.7
9.7
12.8
18.3
32.6
15. 1
16.8
12.2
12.2
12.2
14.8
11.3
12. 5
NSC
5. 1
7.8
NSC
3.5
14.2
8.5
11.0
5.3
10.8
13. 1
13.8
12.8
12. 5
6.7
6.5
6.3
NSC
7.9
11.2
6.9
9.9
10.8
15.4
12.9
NSC
4.9
7. 1
NSC
Period 1
7.6
18.9
7.9
6.6
6.9
21.3
6.6
14.6
9. 1
8. 1
9. 9
4.0
15.9
14.6
25. 1
9.0
5.5
15.2
8.0
NSC
5. 5
10.3
NSC
4.2
14.0
9.1
17.7
10.3
6.2
10. 9
9.9
4.7
15.6
17.7
6.1
6.6
NSC
<2. 1
6.9
5.5
18.9
8.3
14. 1
8.5
NSC
10.3
14. 0
NSC
Pe riod 5
10.9
<2. 1
18.6
2.6
10.7
9.9
6.6
6.9
8.6
5.3
8.9
7.3
5.6
8.5
NSC
5.0
4. 1
2. 4
2.3
11.4
14.3
5.3
NSC
NSC
NSC
NSC
6. 1
2.3
5.3
<2. 1
2.7
<2. 1
8.0
4.6
NSC
NSC
6.5
NSC
NSC
NSC
-------
TABLE A-24 HEMATOCRIT VALUE OF BLOOD SAMPLES
FROM INDIVIDUAL PARTICIPANTS
Participant
I. D.
1
2
3
4
5
6
9
10
11
12
13
14
15
16
18
19
21
29
30
31
32
33
49
50
74
75
76
77
78
83
84
97
98
99
100
101
102
103
113
114
115
116
117
131
141
142
143
144
145
146
157
158
159
160
162
165
166
167
171
172
(continued)
Period 1
43
38
42
47
42
43
40
40
38
46
43
44
39
40
40
43
NSC
42
41
41
42
38
37
41
45
40
38
37
NSC
40
41
48
NSC
NSC
39
44
45
39
38
39
42
39
38
40
40
40
39
36
45
43
48
47
NSC
NSC
42
45
38
41
NSC
NSC
Hematocrits
Period 2
42
NSC
NSC
47
42
42
42
42
39
41
46
44
43
40
39
43
NSC
42
38
38
40
37
41
48
46
42
NSC
NSC
NSC
43
47
48
40
NSC
40
42
41
43
38
40
46
41
39
38
41
39
39
41
47
44
47
42
NSC
NSC
43
45
37
NSC
37
38
Period 5
43
36
44
45
43
46
NSC
42
NSC
45
46
44
42
42
NSC
38
NSC
NSC
41
43
NSC
NSC
41
52
44
43
39
NSC
NSC
44
42
52
38
NSC
42
49
40
43
NSC
NSC
NSC
NSC
NSC
NSC
39
41
39
42
48
44
48
44
39
38
40
45
35
NSC
38
41
357
-------
TABLE A-24 (continued)
Participant
I. D.
177
178
179
180
181
182
183
189
190
191
192
1.93
194
195
196
198
199
201
202
206
207
208
209
213
214
226
227
228
229
230
231
232
233
235
241
242
243
244
254
255
256
257
258
260
262
263
266
267
268
273
275
276
278
284
290
293
294
295
302
303
• 312
(continued)
Period 1
41
39
34
34
32
39
NSC
46
43
37
38
NSC
38
NSC
44
38
39
43
NSC
43
37
43
39
45
40
39
46
38
40
37
41
NSC
45
NSC
42
43
39
40
41
NSC
39
41
52
47
35
40
44
39
41
42
43
36
36
42
35
38
47
45
46
40
44
Hematocrits
Period 2
47
43
42
42
48
39
35
47
43
36
39
38
38
NSC
43
39
38
41
41
43
36
44
38
45
41
39
47
43
42
NSC
38
NSC
43
NSC
39
44
38
37
NSC
42
38
39
47
44
39
37
47
39
47
42
40
37
37
48
42
41
44
NSC
49
44
44
Period 5
47
43
44
41
45
39
35
NSC
NSC
NSC
NSC
NSC
NSC
NSC
43
NSC
41
NSC
NSC
48
37
43
38
44
40
37
44
42
42
NSC
35
38
44
38
43
46
39
41
NSC
NSC
NSC
NSC
NSC
44
41
40
47
41
46
48
38
37
38
NSC
39
NSC
NSC
NSC
52
41
47
358
-------
TABLE A-24 (continued)
Participant
I. D.
313
314
315
316
324
325
330
334
335
336
337
338
339
340
341
342
343
348
349
350
353
354
355
356
359
361
36Z
363
. 364
365
366
367
368
370
372
373
374
375
378
383
384
385
386
387
388
390
391
392
393
394
395
396
397
398
399
400
402
403
405
406
(continued)
Period 1
NSC
38
NSC
42
42
38
47
34
34
38
38
39
39
NSC
NSC
NSC
50
37
NSC
41
41
38
38
36
40
38
38
NSC
48
37
39
42
34
38
43
NSC
41
NSC
49
38
45
35
39
37
38
37
39
36
42
41
37
36
43
41
44
45
44
41
48
43
Hematocrits
Period 2
38
42
38
40
43
42
49
47
41
43
41
NSC
41
NSC
38
42
47
NSC
46
41
39
38
38
37
43
38
40
NSC
48
36
38
40
40
38
43
NSC
46
NSC
NSC
NSC
NSC
36
40
39
38
37
NSC
35
42
43
38
NSC
44
38
44
43
45
42
45
40
Period 5
42
NSC
39
40
42
42
48
48
40
43
41
40
40
37
39
NSC
49
36
NSC
NSC
41
39
41
40
42
37
37
36
48
36
38
42
43
40
38
38
46
38
47
NSC
NSC
40
41
38
36
37
35
39
NSC
NSC
NSC
NSC
NSC
39
42
44
NSC
38
46
38
359
-------
TABLE A-24 (continued)
Participant
I. D.
407
408
41Z
413
414
415
416
417
418
419
421
422
423
424
426
428
429
430
438
441
442
443
445
446
449
450
451
452
453
454
455
457
458
459
460
461
464
465
466
467
500
501
502
503
504
505
506
507
508
509
510
511
512
Hematocrits
Period 1
35
43
45
40
NSC
41
38
44
49
45
39
44
NSC
39
40
38
47
NSC
45
45
38
34
39
36
44
35
NSC
42
45
43
43
38
36
38
39
44
44
33
46
41
32
43
36
38
33
NSC
36
32
44
36
43
NSC
NSC
Period 2
41
45
37
38
NSC
44
37
43
51
45
38
44
47
38
42
41
43
NSC
45
43
38
36
41
35
45
NSC
NSC
40
43
43
42
39
38
42
47
42
48
41
45
39
NSC
NSC
42
43
39
NSC
35
38
46
45
45
NSC
41
Period 3
38
43
NSC
NSC
41
43
41
42
46
43
NSC
43
44
38
39
NSC
NSC
NSC
NSC
42
39
35
39
35
49
36
NSC
NSC
NSC
NSC
43
39
38
41
43
42
49
43
NSC
NSC
34
NSC
NSC
NSC
41
40
34
38
48
42
45
NSC
41
360
------- |