Evaluation of the Health Risks
Associated with the Treatment and
Disposal of Municipal Wastewater and Sludge
Cincinnati Univ., OH
Prepared for
Health Effects Research Lab,
Cincinnati, OH
Mar 81
PB81-175945
U.S. DEPARTMENT OF COMMERCE
National Technical Information Service
-------
EPA 600/1-81-030
March 1981
EVALUATION OF THE HEALTH RISKS ASSOCIATED WITH THE
TREATMENT AND DISPOSAL OF MUNICIPAL WASTEWATER
AND SLUDGE
C.S. Clark1,. H..S. Bjornson2, J.W. Holland2, V.J. E.lia1, V.A. Majeti1,
C.R. Meyer*, W.F. Balistreri3, G.L. Van Meer1. P.S. Gartside1.
B.L. Specker1, C.C. Linnemann, Jr.4, R. Jaffa^, P.V. Scarpino^,
.. K, Brenner5, W.J. Davis-Hoover5, 6.W, Barrett6,
T.S. Anderson6, and D.L. Alexander1
Departments of Environmental Health^, Surgery2, Pediatrics^,
Medicine4 and Civil and Environmental Engineering5
University of Cincinnati
and
Institute of Environmental Sciences6
Miami University
Grant No. R805445
Project Officer
Herbert R. Pahren
Epidemiology 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
-------
NOTICE
THIS DOCUMENT HAS BEEN REPRODUCED
FROM THE BEST COPY FURNISHED US BY
THE SPONSORING AGENCY. ALTHOUGH IT
IS RECOGNIZED THAT CERTAIN PORTIONS
ARE ILLEGIBLE, IT IS BEING RELEASED
IN THE INTEREST OF MAKING AVAILABLE
AS MUCH INFORMATION AS POSSIBLE.
-------
TECHNICAL REPORT DATA
/Please read /ni/rurr.tvti on the rettnt bcforr completing!
1. REPORT NO.
EPA-600/1-81-030
3. RECIPIENT'S ACCESSION NO.
17594 5
4, TITLE AND SUBTITLE
Evaluation of the Health Risks Associated with the
Treatment and Disposal of Municipal Wastewater and
Sludge
S. REPORT DATE
March 1981
6. PERFORMING ORGANIZATION CODE
1. AUTMOR(S)
8. PERFORMING ORGANIZATION REPORT NO.
C. S. Clark, et al.
B. PERFORMING ORGANIZATION NAME AND ADDRESS
Departments of Environmental Health, Surgery, Pediatrics
Medicine and Civil and Environmental Engineering
University of Cincinnati
Cincinnati, Ohio 45267
1O. PROGRAM ELEMENT NO.
CAYB1B
11. C6NTRACT/GRANT N6.
R805445
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency
Health Effects Research Laboratory
26 W. St. Clair St.
Cincinnatir Ohio 45268
13. TYPE OF REPORT AND PERIOD COVERED
Final 2/22/78 - 5/21/80
14, SPONSORING AGENCY CODE
16. SUPPLEMENTARY NOTES
In this study, started in 1977, clinical and serologic evaluations of workers involved in
composting of wastewater treatment plant sludge by the aerated pile method was initiated
to evaluate the potential health effects of exposure to Aspergi1 las f urni gatus and other
viable and nonvtable components of sludge. A health study consisting of analysis of blood
specimens for liver and kidney function parameters, determination of serum and urine bile
acid concentrations, physical examinations, interviews, and air and water monitoring was
conducted of residents exposed to carbon tetrachloride and other toxic chemicals in
drinking water as a result of contamination of domestic wells by a nearby hazardous waste
dump. A study was undertaken to determine if evidence of exposure to hexachlorocyclo-
pentadiene (HEX), hexachlorobicycloheptadiene (HEX-BCH), heptachlorobicycloheptane, and
chlordene could be detected by urine analysis of workers exposed to chlorinated
insecticide substances discharged to the sewer by a pesticide formulator. In order to
assess the potential for health risks associated with the spray irrigation of wastewater,
a clinical and virus serologic evaluation of workers and wastewater and air monitoring
for viruses, pathogenic bacteria, and volatile organic chemicals was conducted. A field
ecosystem study was carried out to evaluate the effects of land application of heat dried
municipal wastewtaer treatment plant sludge (Milorganite) on vegetation, insects and
meadow voles. The potential health effects associated with sludge incineration and
pyrolysis are also presented.
IT.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTiriERS/OPEN ENDED TERMS
COSATI Field/Group
21. NO. OF PAGES
It. DISTRIBUTION STATEMENT
Release to Public
1». SECURITY CLASS fThu Rtpoftl
Unclassified
2O. SECURITY CLASS (Tltu puftl
Unclassified
22. PRICE
f»tm 2220—1 (R»v. 4-77) PKBVIOUI EDITION i» OBSOLETE
-------
DISCLAIMER
This report has been reviewed by the Health Effects Research Laboratory,
U.S. Environmental Protection 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
commercial products constitute endorsement or recommendation for use.
-------
FOREWORD
The U.S. Environmental Protection Agency was created because of increasing
public and governmental concern about the dangers of pollution to the health and
welfare to the American people. Noxious air, foul water, and spoiled land are
tragic testimony to the deterioration of our national 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 the valuable health information generated by
these activities, 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 contains several diversified health related evaluations
relating to the treatment and:disposal of wastewater and sludge. Hopefully, the
information will be of value to those planning, operating, or regulating such
facilities so that health effects may be minimized.
Garner
Director
Health Effects Research Laboratory
lii
-------
ABSTRACT
Information concerning the nature and extent of human health risks involved
in the treatment and disposal of municipal wastewater and sludge is scarce. A
major research program to fill some of the gaps has been undertaken at the
University of Cincinnati, in the Department of Environmental Health, since
1975, which involved collaboration with investigators in other Departments of
the University and at Miami University. The first phase of this effort focussed
on occupational exposure to viruses and bacteria associated with sewer
maintenance and conventional activated sludge wastewater treatment. The
second phase of this effort, starting in 1977, included study of exposure to
different agents: organic chemicals, fungi and endotoxins; and different
processes: composting of wastewater treatment plant sludges, hazardous waste
disposal, the spray irrigation of wastewater, and the land application of
sludge. The various projects carried out during the second phase of the
program are described in this report.
A study of workers involved in composting of wastewater treatment plant
sludge was initiated to evaluate the potential health effects of exposure to
Aspergillus fumigatus and other viable and nonyiable components of sludge in
three eastern cities. The study consisted of clinical and serologic evaluation
of workers five times a year, physical examinations, and environmental
monitoring. The A. fumigatus spores from oropharyngeal and anterior nares
cultures of compost workers were isolated with comparatively high frequency
compared to control groups. Preliminary data also indicated an apparent
antibody response in some of the workers to lipopolysaccharide prepared from
compost samples.
A health study was conducted of the residents of a small community exposed
to toxic chemicals in drinking water as a result of contamination of their
domestic wells by a nearby toxic wastes dump. Testing performed in November
1978 after most use of the contaminated water had ended indicated elevated
levels of the serum enzymes alkaline phosphatase, serum glutamic p'yruvic
transaminase, and serum glutamic oxaloacetic acid in many of the residents.
Two months later many of the persons with abnormal test results had results in
the normal range. Six exposed individuals had slightly enlarged livers
compared to one in the intermediate group. The nonfasting serum concentrations
of the bile acid sulfated conjugates of lithocholate (SLCC) were significantly
lower in the exposed group than in the controls in January 1979. Air samples
collected in some of the homes showed the presence of hexachlorocyclo-
pentadiene.
Workers exposed to chlorinated insecticide intermediates discharged
to the sewer by a pesticide formulator were studied at the North Wastewater
iv
-------
Treatment Plant, Memphis, TN. The study was undertaken to determine if evidence
of exposure to toxic chemicals could be detected in the workers. The study
showed that the wastewater and air samples at the North Plant contained
several contaminants including hexachlorocyclopentadiene (HEX), hexachloro-
bicycloheptadiene (HEX-BCH), heptachlorobicycloheptene, and chlordene.
A number of workers at the North and Maxson (control) plants had HEX and
HEX-BCH in their urine specimens. However, urinary excretions of HEX-BCH
were significantly higher in North Plant workers than in Maxson Plant workers.
Also, urinary excretion of HEX-BCH was found to increase during the work
shift for workers at the North Plant, whereas it decreased during the work
shift at the Maxson Plant. The significance of the presence of these
chemicals in the urine of the workers and the potential long-term health
effects of exposure to these chemicals are not known.
The Muskegon County wastiwater spray irrigation system study consisted
ot clinical and virus serologic evaluation of the workers and environmental
monitoring tor viruses, pathogenic bacteria, and volatile organic chemicals in
wastewater and air samples. Although viruses were present in influent waste-
water and sometimes, in the storage lagoon, no animal viruses were detected
either in the lagoon wastewater just prior to spray application or in the
aerosol samples collected at the aeration basin. Klebsiella was found to
be the most predominant of the gram-negative rod pathogens detected in air
samples. The concentrations of selected volatile organic chemicals detected
downwind of the ;.spray irrigation rigs were, much below the recommended
occupational standards. Clinical evaluation of the workers did not show any
differences in illness and virus isolation rates compared to a control group.
The study showed that the antibody titers to coxsackievirus B5 were sig-
nificantly higher in spray irrigation nozzle cleaners, the workers with the
greatest and most direct exposure to wastewaters. No other differences in
exposed and control workers were detected in levels of antibody to seven other
viruses.
A field ecosystem study was carried out at Miami University, Oxford, Ohio,
to evaluate the effects of land application of heat dried municipal waste-
water treatment plant sludge (Milorganite) on vegetation, insects and
meadow voles. The study concluded that the old-field community is more
stable than the wheat field and better suited for sludge application than
the wheat fields. The survivorship, longevity, percentage of breeding adults
and recruitment rates of voles were not affected by sludge treatment.
Application rates of Pb, Zn, Cd, and Cu were much higher in the sludge-treated
fields than in those treated with a nitrogen and phosphorous-equivalent
commercial fertilizer. There were no differences in tissue concentrations
of Pb, Zn and Cu of meadow voles from any of the fields. However, Cd
concentrations were elevated in kidneys and livers of all voles from
sludge-treated plots for both annual and perennial fields. Although there
were no differences in liver degeneration between voles from the sludge
and commercial fertilizer field plots, a definite effect is present in the
sludge and fertilizer-exposed group compared to the control group.
Reports have been prepared from an evaluation of pertinent literature on
potential health effects from viable and nonviable emissions (EPA-600/
1-81-006) and persistent organics (EPA-600/1-80-025) associated with waste-
-------
water treatment plants and land application sites. The potential health
effects associated with sludge incineration and pyrolysis are presented in
this report. :
This report was submitted in fulfillment of-Grant No. R 805445 by the
Department of Environmental Health, University of Cincinnati, under the
sponsorshop of the U.S. Environmental Protection Agency. This report covers
the period February 22, 1978 to May 21, 1980.
VI
-------
CONTENTS
Foreword ill
Abstract , i v
Fi gures x
Tables..... xli
Acknowledgments . xix
1. Introduct i on ..,. 1
2. Conclusions... 5
3. Recommendations.........., 12
4. Occupational Hazards Associated With Sludge Composting 14
Background 14
Environmental monitoring.... 17
Potential health effects of Aspergillus fumigatus... 17
Elements of compost workers study 19
Population groups selected 19
Methods 21
Clinical and serological 21
Environmental monitoring... 23
Results 24
Populations recruited............................... 24
Air sampling 25
A. fumigatus in study participants 30
Liver function and total immunoglobulin
determination 32
Delayed hypersensitivity to aspergillus antigens 32
C-reactive protein (CRP) 32
Eosinophil count 32
Chest x-ray findings 33
Antibody to A. fumigatus 33
. Antibody to Tipopolysaccharide (IPS) 33
Health effects associated with composting 33
Discussion and conclusion 34
5. An Environmental Health Survey of Drinking Water Contamination
by Leachate from a Pesticide Waste Dump - Hardeman County,
Tennessee 36
Background 36
Methods and procedures 37
Results - November 1978 42
Summary : 49
Results - January 1979 49
Discussion and conclusions 64
vii
-------
6. Evaluation of Worker Exposure to Pesticide Chemicals at a
Memphis, Tennessee, Wastewater Treatment Plant..... 68
Study design. 69
Analytical methods 72
Statistical methods 73
Results.. 73
Urine analysis and illness symptoms. 73
Summary of results of analysis of urine data and
illness symptoms 86
Environmental monitoring 86
Medical 88
Discussion.... ...' 88
7. Viruses in Aerosols and Wastewater at a Wastewater Spray
Irrigation System 94
Site description 94
Methods 96
XM2 biological sampler/collector 96
Collection liquids 96
Measurement of the air velocity of the sampler
air unit 96
Cleaning and sterilization procedures.. 100
Sterility tests 100
Heathkit portable weather station, model ID-1290 102
Sampling procedure . 102
Sample processing and virus assay........ 106
Coliphage assay 107
Bacterial assay 107
• Results and discussion 108
8. Bacterial Aerosol Enumeration and Identification at a Waste-
water Spray Irrigation System . 122
Materials and methods 122
Air sampling 122
Sampling sites 124
Enumeration and gram-negative bacteria isolation
and identification 124
Gram-positive bacteria identification 124
Results 127
Discussion 135
9. Selected Organic Chemicals in Aerosols and Wastewater at a
Wastewater Spray Irrigation System 138
Experimental methods 139
Sampling and analysis procedures 139
Results and discussion 140
10. Sero-epidemiologic Survey of Workers at a Wastewater Spray
Irrigation System 148
Methods 148
Results 149
Volunteers recruited 149
Reported illnesses 149
Virus isolations 149
Virus serology 153
Summary !!!!!!.... 161
viii
-------
11. Effects of Sewage Treatment Plant Sludge on the Structure
and Function of Two Grassland Ecosystems 162
Methods 164
Land treatment 164
Sludge and fertilizer analyses 166
Ecological methods 166
Histopathology 167
Resul ts 167
Metal content of sludge and fertilizer 167
Primary productivity and diversity 170
Arthropods 170
Pathology report on voles 181
Meadow vole populations..... 185
12. Potential-Health Effects from Viable and Nonviable Emissions
Associated with Sludge Incineration and Pyrolysis 191
Introduction , .... 191
Incineration. 194
Air pollution standards......... 196
Emissions from incineration.... 197
Potential health effects of incineration 202
Control of air pollution from incineration 202
Residue disposal 205
Pyrolysis 205
Air pollution from pyrolysis..... 210
Conclusions and recommendation 212
References. 213
Appendix —
A. Monthly illness history...... 227
ix
-------
FIGURES
Number Page
1 Location of contaminated private wells near Hardeman County
toxic waste dump (designated by X) •.„' 39
2 Location of homes of January 1979 control group in relation
to dump site (designated by letter C).... 41
3 Location of Memphis wastewater treatment plants, pesticide
manufacturer and Memphis Light, Gas and Water 70
4 Gas chromatograms of extracts of influent wastewater samples... 87
5 Muskegon County Wastewater Management System No. 1 95
6 XM2 sampler air unit.. 97
7 Schematic of sample gas flow through the XM2 biological sampler/
collector (from the XM2 Operator's Manual)... 98
8 Cross-section of the air collar of the sampler showing the
location of the air flow readings 99
9 Sterile trap system for introducing air into lower tubing
of the sampler 101
10 Location of the XM2 biological sampler/collector during the
sampler runs 103
11 Schematic of aeration basins #1 and #2 showing the location
of the XM2 biological sampler/collector during the specified
days of the sampling runs 104
12 Schematic showing the position of the XM2 biological sampler/
collector in relation to the edge of the wastewater 105
13 Schematic diagram of a six-stage Andersen sampler 123
14 Map of Muskegon Wastewater Management System NO. 1 showing
the air sampling sites 125
15 Flow diagram for enumeration and gram-negative bacteria
isolation and identification scheme 126
-------
FIGURES (continued)
jjumber
16 Flow diagram of the replication and identification scheme
for gram-positive bacteria 128
17 Relative proportions of respirable bacteria identified downwind
of aeration basin in 2.06 cubic meters of air 130
18 Relative proportions of respirable bacteria identified upwind
of aeration basin in 2.01 cubic meters of air 131
19 Relative proportions of respirable bacteria identified downwind
of field rigs in 3.97 cubic meters of air..... 132
20 Relative proportions of respirable bacteria identified upwind
of field rigs in 3.97 cubic meters of air 133
21 Relative proportions of respirable bacteria identified at
other sources in 7.46 cubic meters of air 134
22 Schematic arrangement of individual study plots 165
23 Shannon-Weaver's index in the annual wheat field 173
24 Shannon-Weaver's index in the perennial grass field............ 174
25 Productivity in the annual wheat field.. i..... 179
26 Productivity in the perennial grass field. 180
27 Hydropic degeneration of hepatic cells 182
28 Hydropic and fatty changes in hepatic cells 182
29 More severe hydropic changes in hepatic cells 183
30 Meadow vole population densities in the wheat field 186
31 Meadow vole population densities in the old-field 187
XI
-------
TABLES
Number . Page
1 Scheduling of Elements of Compost Workers Study............ 20
2 Size and Type of Treatment Facilities from Which
Participants Were Recruited. 21
3 Camden Workers: (Age, Race, Sex and Exposure) 26
4 BeltsviHe Workers: (Age, Race, Sex and Exposure) 26
5 Blue Plains Workers: (Age, Race, Sex and Exposure). 26
6 Piscataway Workers: (Age, Race, Sex and Exposure) 26
7 Summary of 1979 Air Monitoring for Respirable Concentrations
of A. fumlgatus and Other Thermophilic Microorganisms
(Colony Forming Units Per Cubic Meter, CFU/M3) 27
8 Summary of 1979 Air Monitoring for Respirable Concentrations
of Fecal Streptococci and Fecal Coliforms (Colony Forming
Units Per Cubic Meter, CFU/M3) 28
9 Summary of 1979 Air Monitoring for Respirable Concentrations
of Coliforms and Total Bacteria (Colony Forming Units Per
Cubic Meter, CFU/M3) 29
10 Distributions and Analyses of Study Participants According
to the Highest Value of Aspergillus fumigatus (AF)
Colonies Observed on Cultures Taken During 1979 31
11 Contaminants Detected in Private Wells in Toone-Teague
Area of Hardeman County, Tennessee 38
12 Age and Sex Profile of Hardeman County Exposed Study Group
and Memphis Control Group (November 1978) 44
13 Hepatic Profile Comparison of Hardeman County Exposed Group
and Memphis Control Group (November 1978) 45
14 Variation in Hepatic Profile With Age and Sex, Hardeman
County - Memphis Comparison (November 1978) 46
xn
-------
TABLES (continued)
Number Page
15 Concentrations of Selected Organic Compounds in Air
Samples (1978) 47
16 Concentrations of Selected Organic Compounds in Water
Samples (1978).... 48
17 Age and Sex Profile of Exposed, Intermediate-Exposed and
Control Groups in Hardeman County Study (January 1979)... 50
18 Hepatic Profile Comparison of Hardeman County Exposed,
Intermediate-Exposed and Control Group (January 1979).... 51
19 Comparison of Hepatic Profile Test Results for Exposed
Participants in Both November 1978 and January 1979
Studies............... 52
20 Transformation and Significant Covariables in the
Statistical Analyses of Liver Function, Bile Acid and
Renal Function Test Data 54
21 Comparison of Mean Bile Acid Levels + Standard Error) in
Fasting and Nonfasting Serum and Urine Samples Collected
From Participants in January 1979 Study...... 55
22 Comparison of Serum Bile Acid Levels in November 1978
Samples With Those in Serum Samples Collected From the
Same Individuals in January 1979 56
23 Comparison of Mean Values (± Standard Error) of Renal
Profile Test Results Between Exposed and Control Groups
Participating in January 1979 Study 57
24 Comparison of Reported Serious Illnesses in Prior Six
Months 58
25 Concentration of Selected Organic Compounds in Water
Samples and Air Samples for Residences of Study
Participants (January 1979) 60
26 Concentration of Selected Organic Chemicals in Water and
Air Samples for Residents of Study Participants
(March 1979) 62
27 Comparison of Water and Air Samples of Residences of
Study Participants (January and March 1979) 65
xm
-------
TABLES (continued)
Number Page
28 Basic Elements of Experimental Design 71
29 Age Distribution of Participants in May and June 1978
Urine Screenings ..... 74
30 Race and Sex Distribution of Participants in May and June
1978 Urine Screenings... 75
31 Age Distribution of Participants in September 1978
Urine Screenings 76
32 Race and Sex Distribution of Participants in September
1978 Urine Screenings. 76
33 Hexachlorobicycloheptadiene (HEX-BCH) Concentrations in
Urine Samples of Memphis Wastewater Treatment Plant
Employees Second Half of Work Shift, May 1978 77
34 Hexachlorocyclopentadiene (HEX) Concentrations in Urine
Samples of Memphis Wastewater Treatment Plant Employees
Second Half of Work Shift, May 1978... 77
35 Hexachlorobicycloheptadiene (HEX-BCH) Concentrations in
Urine Samples of Memphis Wastewater Treatment Plant
Employees, June 1978 79
36 Hexachlorocyclopentadiene (HEX) Concentrations in Urine
Samples of Memphis"Wastewater Treatment Plant Employees,
June 1978 79
37 Prevalence of Self-Reported Symptom During May and June
Surveys 80
38 Average Number of Symptoms Per Worker Plant and Shift,
May-June 1978 81
39 Results of Analyses Urine Collected From Various Groups in
Memphis, Tennessee During July 1978 83
40 Comparison of Results of HEX-BCH Analyses of Urine Specimens
During July 1978 From North Plant Workers and Sampling
Crew With Those From Maxson Plant Workers and Other
Control Workers 84
41 Urine Samples of Wastewater Treatment Plant Employees
Collected at the End of Shift in September 1978 84
xiv
-------
TABLES (continued)
Number Page
42 Prevalence of Self-Reported Symptoms During September
1978 Survey 85
43 Concentrations of Selected Organic Compounds in Influent
Wastewater at Memphis North Treatment Plant, 1978 89
44 Area Air Samples Collected at the Memphis North Plant, 1978.. 90
45 Personal Air Monitoring of Workers at the Memphis North
Wastewater Treatment Plant 91
46 Virus Recovery From Muskegon Air and Water Samples 109
47 Virus Recovery From Muskegon Raw Influent Samples.... 110
48 Comparison of the Coliphage Recovery From Muskegon Air
Samples Using the Soft Agar Overlay Method and Kott's
MPN Method Ill
49 Summary of Weather Conditions at Muskegon During Air Sampling
Runs. .... 113
50 Coliphage Recovery From Muskegon Water Samples Using the
Soft Agar Overlay Method ...... 114
51 Coliphage Recovery From Muskegon Foam Samples Using the Soft
Agar Overlay Method 115
52 Recovery of Bacteria From Muskegon Air Samples 117
53 Standard Plate Counts of Muskegon Water Samples 118
54 Comparison of Standard Plate Counts on Muskegon Foam and
Aeration Basin Raw Influent Samples 119
55 The Advantages and Disadvantages of the XM2 Biological
Sampler/Col lector 120
56 Enumeration of Respirable Colony Forming Units Recovered at
Various Sites 129
57 Initial Air Sampling Surveys at Muskegon County Wastewater
Management System 141
58 Daily Air Concentrations Near Aeration Basins and Associated
Wastewater Concentrations 142
xv
-------
TABLES (continued)
Number
59 Daily Air Concentrations at Aeration Basins and Influent
and Effluent Wastewater Concentrations of the Aeration
Basins 143
60 Daily Concentrations of Selected Chlorinated Organics Near
Spray Irrigation Rigs ....;• 144
61 Air Concentrations of Selected Chlorinated Organics at
Various Locations. ».-.... 146
62 Air Samples Collected at a 30 Million Gallons Per Day Primary
Wastewater Treatment Plant 146
63 Air Concentrations of Selected Organics Collected at Various
Locations at a 300 Million Gallons Per Day Activated
Sludge Wastewater Treatment Plant.. 147
64 Age, Race, Sex and Length of Time on the Job for Spray
Irrigation System and Road Commission Study Participants... 150
65 Age, Race, Sex and Length of Time on the Job for Study
Participants Divided According to Exposure Category 151
66 Comparison of Illness for Spray Irrigation and Road
Commission Workers by Illness Type, Worker Group, and
Exposure Categories 152
67 Comparison of Prevalence of Antibody at Titer Level 2 or
Greater for All Spray Irrigation and Road Commission
Workers During the Month of July 154
68 Comparison of Number of Fourfold or Greater Titer Level
Increases in Spray Irrigation and Road Commission Workers
Using Results From First and Last Serum Collected From
Each Participant 155
69 Comparison of Hepatitis A Antibody Tests for Spray Irrigation
and Road Commission Workers at Both Age 30 and Age 40 156
70 Comparison of Prevalence of Antibody at Titer Level 2 or
Greater for High Exposure Spray Irrigation Workers and
Indoor Road Commission Workers During Month of June 158
71 Comparison of Number of Fourfold or Greater Titer Level In-
creases in High Exposure Spray Irrigation Workers and In-
door Road Commission Workers Using Results From First and
Last Serum Collected From Each Participant 159
xvi
-------
TABLES (continued);
Number Page
72 Comparison of June Coxsackie B5 liter Levels for Nozzle
Cleaners With Those off Various Other Groups of Study
Participants. ........ 160
73 Attributes of Three Stages, of Ecosystem Development Rated
According;to,Their Expected;Capacity for Processing Sewage
Sludge*....... . 163
74, Metal; Content of Sludge, 1978 and 1979 ,., 168
75. Metall Content of Perti 1 izer, 1978 and 1979 169
76 Amounts of the Heavy Metals Applied to the Sludge and
Fertilizer Treated Plots in 1978 and 1979 (Kilograms) 171
77 Shannon Weaver's Diversity Index, Margelef's Richness Index,
and Pielou's Eveness Index in the Annual and Perennial
Field Based on Peak Biomass of Each Species. 172
78 Peak Bioraass, Litter, Standing Dead and Daily Productivity
in the Annual and Perennial Field 175
79 Peak Biomass of Species With a Peak Greater than 5 g nr2 in
the Annual Field 176
80 Peak Biomass of Species With Peak Greater Than 5 g m-2 in
the Perennial Field 177
81 Mean + S.E. for Grain Production in the Wheat Field 178
82 Experimental Groups of Voles and Their Exposure 181
83 Liver Degeneration in Voles on Agricultural Fields 184
84 Liver Degeneration in Voles on Fallow Fields 184
85 Cadmium Concentrations in Voles From the Wheat Field (1978)
and First-Year Old-Field (1979) (jjg/g Wet Weight) 188
86 Cadmium Concentrations in Voles From the Fourth-Year (1978)
and Fifth-Year (1979) Old-Fields Oug/g Wet Weight) 189
87 Characteristics and Potential Impacts of Thermal Reduction
Methods 192
88 Analysis of Trace Elements in Municipal Sludges From 10 U.S.
Cities 195
xvii
-------
TABLES (continued)
Number •.''.: . , . ..,-...•; :/ " . Page
89 Emission Factors for Sewage Sludge Incinerators............. 198
90 Health Effects of Air Pollutants............^..v,,......... 203
91 Solid and Liquid Waste Classification....................... 206
92 Classification of Waste Disposal Sites.................. 207
93 Trace Metal Concentrations in Product Gas Combusion
Products 211
xviii
-------
ACKNOWLEDGMENTS
Volunteers for the compost workers study were recruited in Camden, N.J.,
from the Camden County Municipal Utilities Authority, Mr. John G. Stroka, Chief
Engineer and Ms. Joanne D. Ochs, Compost Superintendent; in Beltsville, MD,
from the operator of the compost facility, the Maryland Environmental
Services, Mr. Clinton R. Albrecht, Chief, Design and Construction and Mr.
Charles Schriver, Compost Superintendent; in Washington, D.C., from the
District of Columbia Department of Environmental Services, Dr. Bailus Walker,
Jr., former Environmental Health Scientist-Administrator, Mr. John Thomas,
Superintendent Blue Plains Water Pollution Control Plant and the TYROC Waste
Management, Inc., Mr. James A. Madison, Executive Vice President, former
operator of compost operations at the Blue Plains Plant and in Piscataway, MD,
from the Washington Suburban Sanitary Commission, Mr. Richard Hocevar,
Director of Maintenance and Operations and Mr. Chris Bratina, Superintendent
Piscataway Water Pollution Control Plant. The cooperation of the above
persons, numerous others associated with their organizations and the vol-
unteers themselves, is sincerely appreciated. The cooperation of Dr. Paul
Marsh, Dr. Pat Millner and Mr. George Wilson of the U.S. Department of
Agriculture, Beltsville, MD, is also appreciated.
The Hardeman County toxic waste dump study and the Memphis water pollution
control plant study were ably supported by Ms. Virginia Boyle, R.N. and Ms.
Sandra Russell, R.N. of Memphis, TN, who were involved in volunteer re-
cruitment, specimen collection and many other activities. Ms. Kathy Hunninen,
M.S. of the TN Poison Control Center helped in the environmental monitoring
activities. The support of the Region IV Atlanta, GA, Office of the U.S.
Environmental Protectection Agency and in particular, Ms. Kitty Taimi, former
Environmental Engineer, is acknowledged. Drs. Larry Lowry and Mitchell Singal
and Mr. Anthony Smallwopd of the National Institute for Occupational Safety and
Health, Cincinnati, Ohio, provided valuable guidance for these two studies.
Workers for the Memphis study were recruited from the Memphis Department of
Public Works, Mr. Joe Taylor, Superintendent and Mr. Tom Anderson, former
Superintendent, Memphis North Plant, Mr. Maynard Stiles, Director and from the
Memphis Light, Gas and Water Division, Mr. W. R. Gardiner and Mr. Bill Taylor.
The support of the volunteers who participated in these studies is appreciated.
Volunteers from the Muskegon County spray irrigation exposure study were
recruited from the Muskegon County Wastewater Management System, Dr. Y. W.
Demirjian, Director and from the Muskegon County Road Commission, Mr. Chuck
Gil Ian, Superintendent.
The capable efforts of Mr. John D. Sedlacek and Mr. Warren G. Taylor,
formerly graduate students at the Miami University, in various aspects of the
ecosystem study is acknowledged.
xix
-------
Special appreciation is expressed to my secretary, Ms. Jane Onslow, for
her very capable support throughout the research described in this report.
xx
-------
SECTION 1
INTRODUCTION
Contact with wastewater, wastewater sludge, and waters receiving such
discharges have long been regarded as a public health risk. Assumptions
concerning these risks have served as a basis for the promulgation of
standards for recreational water quality and for the construction and
operation or facilities for wastewater and wastewater sludge treatment.
However, there is a paucity of documented evidence concerning the nature and
extent of the risks involved (1). The increased pace of construction of new
wastewater and sludge treatment facilities in recent years, the improvement
of existing facilities and the emphasis on the land application of the
residuals produced, lend impetus to the need to evaluate the risks involved.
One approach is to investigate the health conditions of individuals engaged in
wastewater and wastewater sludge treatment and disposal activities.
Since 1975 a major research program has been underway to evaluate the
health risks involved. This research has been centered in the Department of
Environmental Health but has involved extensive collaboration with in-
vestigators elsewhere in the University of Cincinnati and at Miami Uni-
versity. -The initial effort focussed on exposure to viral and bacterial
hazards associated with sewer maintenance and conventional activated sludge
wastewater treatment (2). Beginning in 1977 this effort was expanded to
include exposures to different substances: organic chemicals, fungi and
endotoxins; and different processes: composting of wastewater treatment
plant sludges, hazardous waste disposal and the spray irrigation of waste-
water.
Sewage sludge composting by the windrow method has been practiced for a
number of years by the Los Angeles County Sanitation District. More recently
the development of the aerated pile composting method by the U.S. Department
of Agriculture, Agricultural Research Station at Beltsville, MD, has resulted
in considerable expansion of the use of sludge composting. Currently,
Philadelphia, PA, Camden, N.J., Washington, D.C., Windsor, Ontario, Bangor
and Portland, ME, as well as other smaller cities are engaged in municipal
sludge composting. Many other cities, including New York City, are actively
considering this method. A number of factors are apparently responsible for
the expansion in the use of sludge composting. Among them are the prohibition
of ocean dumping of sludge after 1981, the increase in the amount of sludge
being produced nationally and the growing emphasis on the land application of
wastewater and wastewater sludges. Municipal wastewater contains a variety
or patnogenic microorganisms, many of which are concentrated in the sludge
that is produced. Composting is a thermophilic process and the heat generated
1
-------
helps inactivate the microorganisms present in sludge. However, the heat
generated during composting also encourages proliferation of thermophilic
fungi, in particular, Aspergillus fumiqatus, and aetinomycetes. Aspergillus
fumiqatus has been shown to be one of the most prevalent fungi during the
sludge composting process (3).-. .Aspergi11us fumiqatus 'is 'an opportunistic
pathogen capable of infecting individuals whose defenses have been weakened.
Aspergillus fumigatus can cause severe allergic (asthmatic) reactions in
atopic individuals and aspergillosis and invasive aspergillosis in immuno-
suppressed patients. The workers involved in the compost ing process would be
exposed to potential health risks from exposure to."A. fumigatus and other
viable components of compost. Because they are heat resistant, bacterial
endotoxins are also present in composted sludge. A prospective health study
was initiated in 1979 to determine if workers at municipal sludge composting
facilities are subjected to an increased health risk from exposure to viable
fungal particles, actinomycetes, and endotoxins in addition to the pathogenic
microorganisms that are normally present in municipal wastewater and sludge.
A description of this study, performed under the direction of H.S. Bjornson,
M.D., Ph.D., of the Department of Surgery, and a preliminary assessment of the
results obtained in 1979 are presented in Section 4: Occupational Hazards
Associated with Sludge Composting.
Improper disposal of hazardous chemical waste is increasingly being
recognized as a source of potential human health hazard. A health study of the
residents of a small community exposed to toxic chemicals in drinking water as
a result of contamination of their domestic wells was initiated in 1978. The
community under consideration is in Hardeman County, TN. Residents of this
community began noticing unpleasant chemical tastes and odors from their
water. Preliminary environmental monitoring indicated that the residents
have been exposed to carbon tetrachloride (CC14) and other chlorinated
organic contaminants from their private water supply wells and possibly in
ambient air. The source of these contaminants in groundwater is most probably
from leachates from a chemical waste landfill formerly used by a Memphis, TN,
pesticide formulator which manufactures chlorinated insecticides. In No-
vember 1978, the residents of the affected community were advised by U.S.
Environmental Protection Agency (EPA), Region IV, Atlanta office, to stop
using the water because monitoring by EPA showed high concentrations of CC14
(up to 10,000/ug/L) in the water samples. A study to determine if the
residents of the Toone-Teague community, Hardeman County, TN, showed any
detectable health effects from exposure to toxic organic chemicals in their
drinking water was conducted during the Fall and Winter of 1978-1979. W. F.
Balistreri, M.D., of the Department of Pediatrics, assisted members of the
Department of Environmental Health with this study. • Section 5: An
Environmental Health Survey of Drinking Water Contamination by Leachate from.
a Pesticide Waste Dump - Hardeman County, Tennessee, is a description of this
study and its results.
During the prospective seroepidemiolpgical study of the bacterial and
viral disease risks associated with municipal wastewater treatment (2), an
opportunity developed to investigate exposure to toxic organic chemicals
emitted from the wastewater during the treatment process at one of the plants
in the study. Workers at this plant, the Memphis North Wastewater Treatment
Plant, had been complaining in early 1978 of acute symptoms of respiratory
-------
distress that they associated with periods of intense chemical odor at the
plant. Other symptoms noted were dizziness, headache, and irritation of the
eyes, nose, throat, lungs and skin. The treatment plant is located near a
manufacturer that produces and utilizes several chlorinated organic inter-
mediates for the formulation of flame retardants and chlorinated insecticides
(notably isodrin, endrin, chlordane and heptachlor). Chemical waste from
this manufacturer is discharged into a sewer that flows to the Memphis North
Wastewater Treatment Plant. During 1978 exposure to several chlorinated
organic compounds at this treatment plant was assessed by evaluating urinary
excretion, illness symptoms and air and wastewater. A description of this
study is presented in Section 6: Evaluation of Worker Exposure to Pesticide
Chemicals at a Memphis, Tennessee, Wastewater Treatment Plant.
As a result of legislative actions, such as the 1972 Clean Water Act and its
1977 Amendments, land application of municipal wastewater and sludge is
becoming increasingly popular as an alternative means to the more con-
ventionally used disposal methods, such as ocean and surface water dumping,
and incineration. Land application represents a recycling process in which
water and plant nutrients are returned to the soil. However, a large variety
of potential disease-causing microorganisms are present in municipal waste-
water and conventional wastewater treatment does not completely remove the
microorganisms. Spray or sprinkler application is one of the commonly used
methods of land application of wastewater, but spray application also
promotes aerosol formation. The volatile organic chemicals and micro-
organisms present in wastewater may become airborne during spray application.
Aerosolization of volatile organic chemicals and microorganisms is also
favored near aeration basins of the activated sludge treatment units and
trickling filters at wastewater treatment plants. The workers involved with
the application of the wastewater and sludge may be exposed to microorganisms
and volatile organic chemicals in aerosols. In order to assess the potential
health risks of workers from exposure to volatile organic chemicals and
viruses in aerosols, the ambient air environment and the workers at the
Muskegon County Wastewater Management System, Michigan, were evaluated. This
evaluation is described in the following four sections of this report:
Section 7 (Viruses in Aerosols), Section 8 (Bacterial Aerosols), Section 9
(Organic Chemical Aerosols) and Section 10 (Virus Serology Survey of
Workers). Sections 7 and 8 were performed under the direction of P. S.
Scarpino, Ph.D., of the Department of Civil and Environmental Engineering,
and Section 10 under the direction of C. C. Linnemann, Jr., M.D., of the
Department of Internal Medicine.
To date most studies of the potential harmful effects of land application
of wastewater and sludge have been concerned with uptake and accumulation of
heavy metals by food crops or forage plants (4,5). Some laboratory feeding
experiments have demonstrated uptake and accumulation of heavy metals by
consumption of sludge (6,7). It has not been adequately shown in field
studies whether animals resident on sludge-treated land concentrate heavy
metals in their tissue and whether the animals are adversely affected by
sludge. A field ecosystem study was carried out at Miami University, Oxford,
OH, under the direction of 6. Barrett, Ph.D., Institute of Environmental
Sciences, to evaluate the effects of land application of municipal sludge on
vegetation, insects and meadow voles (Section 11).
-------
Reports have been prepared from an evaluation of pertinent literature
on potential health effects from viable and nonviable emissions (EPA-600/-1-
81-006) and persistent organics (EPA-600/1-80-025) associated with wastewater
treatment plants and land application sites. The potential health effects
associated witn sludge incineration and pyrolysis are presented in this
report (Section 12).
-------
SECTION 2
CONCLUSIONS
Occupational Hazards Associated With Sludge Composting
1;. Aspergillus futtngatus concentrations in air at the Piscataway Plant and
at areas of the Blue Plains Plant remote from the compost pit were
clearly lower than levels reached at the Beltsville and Blue Plains.
"composting sites. Overall, about six percent of the A. fumtgatus
colonies were /ound on Andersen stage six plates representing samples
thought to be small enough to reach the alveoli.
2. Fecal streptococci were detected in air samples at each of the areas
where they were measured and were generally higher at compost sites.
3. Results of the environmental monitoring demonstrated that workers at
each of the compost sites were exposed to markedly elevated numbers of
A. fumigatus spores. This observation was further supported by the
comparatively high frequency of isolation of A. fumigatus from
oropharyngeal and anterior nares cultures of compost workers when
compared to the intermediate-exposed and control groups.
4. Preliminary analyses of the highest results obtained for each
participant on tests for the enzymes serum glutamic oxaloacetic
transaminase (SGOT), serum glutamic pyruvic transaminase-(SGPT),
measures of liver function, and levels of total IgG, IgE, and IgM, did
not reveal any differences among exposure groups at any of the sites.
5. Preliminary analysis of the highest C-reactive protein (CRP) value for
each individual was performed en two groupings of the data: (1) ^1
mg% and> 1 mg% and (2) positive and nonreactive. The only significant
difference was found among Beltsville workers. Four of nine compost
workers had positive CRP values conpared to none of eight intermediate-
exposed workers.
6. Preliminary analyses of the highest absolute eosinophil count for each
of the participants did not reveal any differences among the exposure
groups at the various sites, separately or combined.
7. Chest X-rays were obtained from 74 participants and reviewed by a
radiologist at the University of Cincinnati. Of these, one compost
worker's x-ray exhibited an abnormality compatible with an occupa-
tional ly-related disorder.
-------
8. Specific IgG directed towards A. fumigatus, as determined by the
enzyme- linked immunosorbent assay (ELISA-), was compared by exposure
group for each site separately. At no site did the compost workers
..exhibit statistically higher values.
9. Titer levels of antibody directed against lipopolysaccharide (LPS) pre-
pared from site-specific compost were not found to be higher among
Camden compost workers than in control workers. However, in Beltsville
and Blue Plains antibody titers among compost workers were higher than
in their corresponding lower exposure groups.
10. One worker in the study, employed at a compost site, was found to have
a local infection involving his right ear. This infection, caused by
A_. niger gradually progressed over a period of several months. Despite
treatment, there was erosion of 70% of the right tympanic membrane.
An Environmental Health Survey of Drinking Water Contamination by Leachate
From a Pesticide Waste Dump - Hardeman County, Tennessee
11. The residents of Toone-Teague Road in Hardeman County, TN, were exposed
-to carbon tetrachloride and other chlorinated organic chemicals in
their drinking water and. possibly in ambient air.
12. The concentrations of carbon tetrachloride, a suspected carcinogen,
found in the contaminated well water (up to 18,700^ug/L) were far
above the level of 2.6/jg/L that has been proposed by the-4J.S.
Environmental Protection Agency as a safe level for water.
13. Concentrations of carbon tetrachloride in the contaminated wells were
of the order of 5000jug/L. Assuming a consumption of one liter per
day, the individuals using the contaminated water had a dose of carbon
tetrachloride of about 5 mg/day.
14. There is a possibility of future contamination of the much deeper
artesian aquifer, which is used at another location by Memphis, TN.
15. Air samples collected in some of the Hardeman County homes showed the
presence of HEX, carbon tetrachloride and tetrachloroethylene.
16. Results from the initial screening in November 1978 revealed that the
concentrations of the serum enzymes alkaline phosphatase SGPT and SGOT
were elevated in the group of residents who had previously consumed the
contaminated water when compared to the control group. The differences
were statistically significant (p /, 0.05) and appear to be associated
with the ingestion of well water contaminated by leachate from a toxic
waste landfill. Two months later many of the persons with abnormal
test results had results in the normal range.
17. Analysis of urine samples for selected organic compounds (HEX, HEX-BCH,
heptachlorobicycloheptene, chlordene and heptachlor) did not show the
presence of any of these chemicals in the exposed group.
-------
18. No evidence was found for carbon tetrachloride in the urine specimens
of a select group of individuals judged to have relatively high potential
for exposure to carbon tetrachloride.
19. During the January 1979 survey, six individuals from the exposed group
and one from the intermediate-exposed were found to have enlarged livers.
20. The mean nonfasting serum bile acid, sulfated conjugates of lithocholate
(SLCC), was significantly lower in the exposed groups than in the control,
21. For those persons in the exposed group who were tested in both November
1978 and January 1979 nonfasting serum concentrations of SLCC and
cholyglycine (CG) were significantly higher in November 1978 than in
January 1979.
22. The residents of Toone-Teague Road, of Hardeman County, TN, have ex-
perienced acute symptoms such as skin and eye irritation, weakness in
the upper and lower extremities, upper respiratory infection, shortness
of breath and severe gastrointestinal symptoms including nausea. These
symptoms usually subsided once the consumption of the contaminated
water was stopped.
23. The potential long-term health effects of exposure to these chemicals
in drinking water is not known.
Evaluation of Worker Exposure to Pesticide Chemicals at Memphis, Tennessee,
Wastewater Treatment Plant
24. Influent wastewater from the North Plant, and air samples collected at
the wet well and grit chamber of the North Plant, contained several
contaminants including HEX, HEX-BCH, heptachlorobicycloheptene and
chlordene.
25. Influent wastewater and air samples obtained at the grit chamber of the
Maxson Plant, the control plant, did not contain the type of substances
found in the samples obtained at the North Plant.
26. Many workers (North and Maxson Plant) had HEX or HEX-BCH in their urine
specimens in May and/or June 1978. However, urinary excretions of
HEX-BCH were significantly higher in North Plant workers than in Maxson
Plant workers in both May and June 1978.
27. The presence of the organic chemicals in the urine samples of Maxson
Plant workers may possibly be due to (a) intermittent discharge into
the sewer system by either a user of these chemicals or a waste hauler,
(b) discharge from a buried dump site containing these chemicals, or
(c) ambient air pollution in the Memphis area.
28. Urinary excretion of HEX-BCH was found to increase during the work
shift for workers at the North Plant in the June 1978 survey, whereas
it decreased during the work shift at the Maxson Plant. The percent of
-------
North Plant urine samples with HEX-BCH concentrations greater than 3
./jg/L increased from 7 to 41 during the work shift while in Maxson Plant
samples the percent decreased from 9 to 0, during the June 1978 screening.
29. The .significance of the presence of these chlorinated organic chemicals
in the urine of workers is not known.
30. The workers at the North Plant experienced acute symptomatology
potentially related to the toxic agents in the influent. The symptoms
decreased or disappeared when the workers were removed from the
contaminated environment.
31. The potential long-term health effects of exposure to these chemicals
is not known.
Viruses in Aerosols and Wastewater at a Wastewater Spray Irrigation System
32. No animal viruses were detected in air samples collected at the Muskegon
County Wastewater Management System using the Army prototype XM2 Bio-
logical Sampler/Collector.
33. Animal viruses were detected in raw influent samples but decreased in
concentrations as the wastewater was aerated and stored in the lagoons.
None were detected in wastewater at the pump station just prior to
distribution to the spray irrigation rigs.
34. Using the centrifugation-filtration method all raw influent samples
were found to contain viruses, ranging from 50 to 400 plaque-forming
units/liter.
3b; Coliphage were removed from air samples at the aeration basins in con-
centrations ranging from 0-9 per cubic meter of air. E_. cpli 13706
coliphage were recovered more often than the other two tested, 15597
and 11303.
36. Kott's MPN method was found to be more sensitive and consistent in de-
tecting low levels of coliphage in air samples than the soft agar over-
lay method.
37. Total bacteria concentrations at the aeration basins ranged up to 7,100
colony forming units per cubic meter of air.
Bacterial Aerosols Enumeration and Identification at a Wastewater Spray
Irrigation System
38. The aeration basin was a source of the following bacteria:
Aeromonas hydrophi la
fc'scnericma coll
Enteropacter agglomerans
other Enterobacter spp.
-------
Klebstella spp.
Pasteurella spp.
Sa1monelTa~spp.
Serratia~spp.
39. -The field rigs were a source of:
Aeromonas hydrophila
Enterobacter aggVomerans
other Entrobacter spp.
Klebsiella spp..
Pasteurella spp.
Pseudomona spp.
SerratiaTpp.
Staphylococcus aureus
40. Four of the bacteria identified are in the Class 2 group of the Center
for Disease Control's "Classification of Etiologic Agents"*
Klebsiella spp.
PasteurefTa spp.
SalmpnelTa~SPP.
Staphy1ococcus aureus
(By use of API-20E system for identification 19 of the 32 Klebsiella
spp. were determined to be Klebsiella pneumoniae.)
41. The quantity of bacteria in the air downwind of treatment sources at
the Muskegon wastewater site contained higher total numbers of bacteria
and higher percentages of gram-negative bacilli, fecal-indicator bacteria,
and pathogenic bacteria than upwind air.
42. The mean respirable concentration of total airborne bacteria found one
meter downwind of the aeration basin at the Muskegon wastewater site
was 2800 Colony Forming Units (CFU) per m3 which was significantly
higher than that found 18 meters downwind of the spray irrigation rigs
(i.e., 700 CFU per m3). Concentrations one meter upwind of the aeration
basin and 18 meters upwind of the field rigs were 490 CFU/m3 and 660
CFU/m3, respectively.
Selected Organic Chemicals in Aerosols and Wastewater at a Wastewater Spray
Irrigation System
43. Concentrations of trichloroethane, trichloroethylene and tetrachloro-
ethylene at the Muskegon County Spray Irrigation System at the downwind
edge of the aeration basins ranged up to 90>ug/L, 73jug/L and 46>ug/L,
respectively. Adjacent to the spray irrigation rigs the highest con-
centrations reached for the same compounds were 2.7, 9.3 and 8.6,
respectively. All of these concentrations were well below the 8-hour
*Class 2 agents are those of ordinary potential hazard, and includes agents
which may produce disease of varying degrees of severity.
-------
occupational standards of 45,000/jg/L, 535,000/jg/L and 670,000 jug/L,
respectively.
44. Chloroform was not detected in the air adjacent to the spray irrigation
rigs but was detected downwind of the aeration basins at concentrations
ranging up to 202.yug/m3 (8-hour occupational limit 50,000>ug/m3). In
the scrubber house chloroform was detected at 3380>ug/m3.
45. Comparing upwind and downwind air concentrations with influent and
effluent wastewater concentrations at the aeration basins showed that
significant concentrations of the volatile substances measured are
stripped from the wastewater during the aeration process.
Seroepidemiologic Survey of Workers at a Wastewater Spray Irrigation System
46. Illness and virus isolation rates were not significantly different in
the study group of workers engaged in the spray irrigation of waste-
water compared to a control group of road commission workers.
47. Antibody titers to coxsack-ievirus B5 were significantly higher for one
subgroup of wastewater workers, the spray irrigation nozzle cleaners.
This suggests that there may be a risk of viral infection only in those
with the greatest and most direct exposure to wastewater.
48. The prevalence of Hepatitis A antibody correlated with age, as would be
expected in normal populations, and there was no increase in the
prevalence of Hepatitis A antibody in those exposed to spray irrigation.
49. Antibody titers to poliovirus 1, 2 and 3, coxsackievirus B2 and echovirus
7 and 11 were not different between the wastewater and road commission
worker groups.
Effects of Sewage Treatment Plant Sludge on the Structure and Function of
Two Grassland Ecosystems
50. When heat-dried Milwaukee, Wisconsin wastewater treatment plant sludge
(Milorganite) and a commercial fertilizer were applied so that the
amounts of added nitrogen and phosphorous were equivalent, the ratio of
the amount of the following heavy metals in Milorganite to that in
fertilizer was 495 for lead, 167 for cadmium, 953 for copper and 280
for zinc in 1978. In 1979 the ratios were 209, 205, 1163 and 334,
respectively.
51. The Shannon-weaver diversity index was lower in the wheat fields re-
ceiving Milorganite than in the wheat fields receiving commercial
fertilizer and the controls. No significant differences were observed
in this index in the o>d grass fields suggesting that the perennial
community is more stable than the annual field and better suited for
application of sludge than the annual field.
10
-------
52. There was a significant increase in productivity in both the perennial
and annual fields in those receiving fertilizer and sludge treatments,
however, the increase was larger and occurred earlier in the
fertilizer-treated fields. Estimates of net primary productivity in
all of the annual fields were more than 50 percent higher than those
for the perennial fields. The treatment effect on plant community
structure was greater in the annual field than in the perennial field,
suggesting again that the perennial fields are more stable and better
suited for sludge application than annual fields.
53. For arthropods the fertilizer and sludge plots exhibited consistently
higher species richness values than did the control plots in both types
of fields. The opposite was true with respect to apportionment values.
54. No pathologic alterations were seen in the lungs and kidneys of the
meadow voles from the fertilized, sludge-treated and control plots of
both the perennial or annual fields.
55. For both the annual and perennial fields the incidence of liver de-
generation in the meadow voles did not differ between the fertilized
and sludge treated plots but was lower in the control plots.
56. Meadow voles from the sludge-treated annual field showed a more severe
liver degeneration than those from the sludge-treated perennial fields.
57. Vole population densities were affected by treatment only in the first
year wheat fields where sludge treatment plots had higher densities
than plots with any of the other treatments in the two years.
58. Survivorship, longevity, percentage of breeding adults and recruitment
rates were not affected by sludge treatment.
59. There were no differences in tissue concentration of Pb, Zn and Cu of
meadow voles from any of the fields. However, Cd concentrations were
elevated in kidneys and livers of voles in both the annual and perennial
sludge-treated fields.
Potential Health Effects From Viable and Nonviable Emissions Associated With
Sludge Incineration and Pyrolysis
60. A literature evaluation indicated that incineration and pyrolysis
processes may lead to air pollution from volatilization of heavy metals
and incomplete combustion of organic chemicals, respectively. The air
pollution can be controlled by the use of appropriate control measures
in both incineration and pyrolysis processes and also by afterburning
in case of pyrolysis.
11
-------
SECTION 3
RECOMMENDATIONS
1. Compost workers should be observed over an extended period of time to
reveal abnormalities which may be related to the length of exposure.
2. The preliminary data on antibody response of the compost workers to
lipopolysaccharide prepared from compost samples suggests that it may
be advisable to take precautions to reduce exposure such as the use of
respirators by compost workers and periodic water spraying of the
compost sites to reduce dust.
3. A method should be developed to quantify the presence of endotoxin in
compost and in the air.
4. Long-term medical follow-up of the residents of Toone-Teague Road,
Hardemah County, TN, should be initiated.
5. Necessary precautions should be taken, if possible, to prevent the
contamination of the artesian aquifer by the toxic chlorinated organic
chemicals since it is used as a water supply for the City of Memphis.
6. The closure plan for the Hardeman County dump site should be designed
to prevent an increased exposure, to nearby residents and to those
downstream and downwind, to contaminants associated with the dump.
7. Industrial discharge of toxic chemical waste into municipal sewerage
systems should be controlled and monitored.
8. A follow-up study of the Memphis workers involved should be conducted
to determine if urinary excretion of HEX and HEX-BCH remains at the low
levels detected in September 1978 and if any health effects are
apparent.
9. The XM2 Biological Sampler/Collector should be used for a sampling
period of at least several days to determine if animal viruses can be
detected in aerosols sampled by this method.
10. The virus content of solid material at the bottom of the lagoons and
aeration basins at the Muskegon spray irrigation site should be
determined to see if future disposal of the solid material would be a
problem.
12
-------
11. The Kott's MPN method is recommended for use when coliphage levels are
expected to be very low.
12. Because of problems with the XM2 sampler and sterilization and arcing
problems in other large volume samplers, an improved large volume
biological air sampler should be developed.
13. Resistant aerosolized bacteria such as Klebsiella should be explored as
possible indicators of bacterial air contamination from fecal sources.
14. An examination of the serology of workers at the Muskegon spray
irrigation site for antibodies to Klebsiellae should be initiated as
soon as possible.
15. Routine air monitoring for selected organic compounds at the Muskegon
spray irrigation site should be performed downwind of the aeration
basins and in the scrubber house.
16. The observation of an apparent increased risk in nozzle cleaners at the
Muskegon spray irrigation site should be confirmed. The number of
available nozzle cleaners is small, but a serologic survey of former
nozzle cleaners could be conducted.
17. The hypothesis that wastewater workers may be at risk of Legionnaire's
disease could be evaluated further by testing for antibody to
Legionella pneumophila which has been related to blue green algae.
Wastewater irrigation workers may be exposed to wastewater with higher
algal content than ordinary wastewater treatment plant workers.
18. For minimizing effects on small mammal populations mid-successional
old-fields should be used as sites for the land application of sludge
rather than early successional stages or agricultural fields.
19. Research should continue to more completely characterize the effect of
sewage sludge application on mammal populations, especially to evaluate
if Cd concentrations tend to increase in the kidneys and livers of
animals functioning within sludge-treated plant communities.
20. Afterburning should be used in pyrolysis process to assure complete
combustion of organic chemicals in sludge.
13
-------
SECTION 4
OCCUPATIONAL HAZARDS ASSOCIATED WITH SLUDGE COMPOSTING
Workers engaged in the handling of municipal wastewater treatment
plant sludge are exposed to a variety of biological, physical and chemical
factors which may present potential health risks. These risks depend upon
the nature of the area served by the treatment plant and the type and con-
dition of sludge treatment process(es) in use.
This section will focus on potential health effects of exposure to
microbial agents associated with the composting of municipal wastewater
treatment plant sludge. Following a brief review of the literature, pre-
liminary results will be presented of an ongoing health study of workers
engaged in municipal sludge composting at several sites in the Eastern U.S.
BACKGROUND
Sewage sludge composting by the windrow method has been practiced for
a number of years by the Los Angeles County Sanitation District. More
recently the development of the aerated pile composting method by the U.S.
Department of Agriculture, Research Station at Beltsville, Maryland, has
resulted in considerable expansion of the use of sludge composting. Cur-
rently, Philadelphia, Pennsylvania; Camden, New Jersey; Washington, D.C.;
Windsor, Ontario; Bangor and Portland, Maine; as well as other smaller
cities are engaged in municipal sludge composting by the aerated pile
method. Many other cities, including New York City, are actively con-
sidering this method of sludge treatment. A number of factors are respon-
sible for the expansion in the use of sludge composting. Among them are
the prohibition of ocean dumping of sludge after 1981, the increase in the
amount of sludge being produced nationally, and the growing emphasis on the
land application of wastewater and wastewater sludges.
Municipal wastewaters may contain a variety of potentially pathogenic
microorganisms. Conventional wastewater treatment does not completely
destroy these microorganisms, many of which are concentrated in the sludge.
Therefore, sludge should be stabilized by a method such as composting,
prior to its application on land. Composting is a thermophilic aerobic
decomposition process. The heat generated during the composting process
has been shown to effectively reduce the numbers of viable microorganisms
present. Two types of composting processes are currently in use -- windrow
and forced aeration pile systems (8,9). The windrow system consists of
long, low piles which are aerated by periodic turning. The forced aeration
pile system consists of a stationary compost pile constructed over a network
14
-------
of porous pipe attached to a blower which draws air through the pile.
Temperatures in the range of 55°-65°C are usually attained during the com-
posting process (8) provided the mixing or aeration is efficient. One of
the most important objectives of composting is to obtain these high uniform
temperatures throughout the system for sufficient duration so as to penetrate
the entire mass. When this objective is fulfilled the composting process
will inactivate most microorganisms including viruses.
The heat generated during composting results in temperatures ideal for
the proliferation of many thermophilic microorganisms such as actinomycetes,
murcorales and in particular, Aspergillus fumigatus. Therefore, compost
workers potentially may be exposed not only to the enteric pathogenic micro-
organisms present in raw sludge (also referred to as primary pathogens
since they are capable of initiating an infection in an apparently healthy
individual) but also to the thermophilic fungi and actinomycetes that pro-
liferate during composting. Some of the thermophilic fungi are capable of
infecting individuals whose defenses have been compromised (these.fungi are
referred to as secondary pathogens). The dust at composting sites may also
contain significant quantities of lipopolysaccharide (IPS or endotoxin)
derived from viable and nonviable gram-negative microorganisms which are
present in sludge. In addition, the fungus Aspergillus flavus which pro-
duces alfatoxin, one of the most potent known human carcinogens, may increase
in numbers during composting and the aflatoxin produced by this fungus may
present an additional hazard for compost workers. Detroy et al. (10), have
shown that the optimal conditions for aflatoxin production (i.e., moisture
content, humidity, temperature, incubation time, aeration and nitrogen and
carbohydrate content) are simmilar to conditions present in portions of the
aerated compost pile (11). It has also been shown that aflatoxin is not
destroyed by temperatures of 60-80°C and therefore would not be detoxified
by the temperatures generated during composting (10). The long term health
effects of chronic exposure to the combination of. microbial pathogens,
microbial toxins and IPS which may be present in the work environment at
composting sites have not been investigated.
Rylander et al., studied various parameters of acute and chronic in-
flammation in workers exposed to dust arising from heat treated sludge at a
sewage treatment plant in Gothenburg, Sweden (12,13,14). These investigators
observed a significant elevation in immunoglobulins (IgG, IgM, and IgA),
leukocytes and platelets in workers at the sewage treatment plant as
compared to a group of age matched control workers at a neighboring oil
refinery. In addition, elevated levels of C-reactive protein and fibrinogen
degradation products were observed in significantly greater numbers of the
sewage treatment plant workers as compared to the control population.
Workers at the sewage treatment plant were also reported to experience the
following clinical symptoms, apparently related to heavy dust exposure:
a) fever, b) purulent discharge from the eyes, c) diarrhea, and d) fatigue.
The investigators postulated that the serologic changes and clinical
symptoms observed in the sewage treatment plant workers may be related to
endotoxin which is present in the dust arising from the heat treated sludge.
They expressed concern regarding the potential health risks associated
with the chronic exposure to endotoxin containing dusts in the work en-
vironment.
15
-------
Rylander et al., also studied the airway immune response to inhaled
endotoxin in laboratory animals (15). Rats were exposed to an aerosolized
endotoxin solution (Escherichia coli 025;B6) daily for 10 days. The dose
of IPS deposited in the lungs of each rat was estimated to be 0.3>Ajg per
day. After the 10-day exposure, elevated levels of IgG, IgM and IgA antibodies
specific for IE. coli 025:B6 endotoxin were demonstrated in the sera of the
exposed rats. Bronchial washings from the exposed rats contained IgG and
IgA antibodies directed against the £. coli 025:B6 endotoxin. In addition,
the bronchial lavage fluid was also observed to contain an increased number
of polymorphonuclear leukocytes (PMN) following endotoxin challenge. At
cessation of exposure, the number of PMN's in the bronchial lavage fluid
decreased and reached values comparable to those observed in control rats
within three days. These observations suggested that inhalation of
endotoxin is capable of producing an acute inflammatory response in the
lungs of laboratory animals and that long term exposure to dusts containing
endotoxins may lead to pathologic changes in the pulmonary parenchyma
caused by the persistent inflammation. McGuire et al. (16) recently re-
ported acute pulmonary inflammation in rhesus monkeys induced by in-
travenous infusion of purified bacterial LPS. These investigators have
demonstrated the presence of an enzyme which cleaved components of the
contact and complement systems in the lungs of the LPS challenged animals.
Rylander and his colleagues have extended their studies of the immune
response to inhaled endotoxin to include workers exposed to cotton dust
in cotton mills in England and Sweden (14,15). Workers with a history of
exposure to cotton dust and subjective pulmonary symptoms of chest tight-
ness were shown to have, in their nasal secretions, elevated levels of
and IgA antibodies specific for antigens prepared from a gram-negative
bacteria isolated from cotton plants being processed in the mills. No
elevation of IgG or IgA antibody levels with specificity for the antigen
preparations employed were observed in nasal secretions obtained from a
control population of workers with no history of exposure to cotton dust.
Rylander postulated, based on his human and animal data, that chronic in-
halation of endotoxin with the accompanying inflammatory response in
the lungs, may play a critical role in the development of Byssinosis.
Dutkiewicz (17,18) conducted an immunologic survey of grain handlers
in Poland who were exposed to high concentrations of the gram-negative
bacterium Erwinia herbicola (synonym Enterobacter agqlomerans) in their
work environment. Workers exposed to dusts containing large numbers of £•
herbicola (5 x 104 to 1 x 105 cfu/m3) had a significantly higher frequency
of serum precipitating antibody directed against E. herbicola antigen
preparations than unexposed individuals. Furthermore, intradermal skin
testing with antigens prepared from E_. herbicola revealed a higher incidence
of positive skin reactions in workers exposed to E_. herbicola than in un-
exposed individuals. Grain handlers exposed to dust containing E_. herbicola
and who complained of respiratory symptoms such as cough, dyspnea, amd"
wheezing, were shown to have a higher incidence of positive skin and
precipitin reactions to £. herbicola antigens than asymptomatic exposed
workers. This study provides further evidence to suggest that exposure to
gram-negative LPS by inhalation is capable of inciting a systemic immune
16
-------
response and that it may play a primary role in the development of chronic
pulmonary disease in exposed individuals.
Lundholm and Rylander (19) recently reported on a study of 11 workers
at a plant where municipal sewage sludge and household garbage were
crushed, miled, and allowed to compost for six months. Gram-negative
bacteria were reported to be present in large numbers where garbage was
loaded onto the conveyor belts (28,000 cfu/m3) and even higher near the
mill outlet (25,000-500,000 cfu/m3). Elsewhere at the plant they ranged
from an average of 330 to 11,000 cfu/m3. At the central water treatment
plant levels averaged 20 cfu/m3. Six of the 11 compost plant workers
reported symptoms of nausea, headache, fever or diarrhea compared to
only two of the 41 water treatment plant employees who served as controls.
Environmental Monitoring
Mi liner-, Bassett and Marsh (20) recently reported on their monitoring
of airborne spores of A. funngatus; at two compost sites at Beltsville,
Maryland and at the Blue Plains Wastewater Treatment Plant in Washington,
D.C. By volumetric sampling they were able to determine that the dis-
persal of the spores from composting sewage sludge and wood chip
mixtures behaved as a Gaussian plume. A major source of airborne spores
was found to be the front-end loader which was estimated to aerosolize
4.6 x 10° A. fumigatus spores/second when moving and dropping compost.
The sampling instrument used was the six-stage Andersen sampler with the
sampling orifice directed into the wind and.with the top cone removed to
collect more nonrespirable particles J21). Other investigators^) in- .
eluding us, (23) have neither removed the top cone nor .tilted the sampler in
their sampling procedure. At the Beltsville compost site airborne con-
centratTbns of A. furni gatus between three and 30 meters*downwind of a
compost pIIe~beTng ag i ta ted by a front-end;loader were reported to range
from 1400 to 3100 cfu/m3. Fifteen minutes after agitation, levels were not
above 39 cfu/m3.
Potential Health Effects of Aspergillus fumigatus
Aspergillus species are ubiquitous in the environment of most countries
of the world.The fungus grows well on a variety of substrates, including
stored hay or grain, decaying vegetation, soil, and dung. A. fumigatus grows
well at 45°C or even higher, making it one of the most common microorganisms
found in compost sites (3). A_. fumigatus, and others of the A. species,
have been shown to be capable of causing disease in both normal and compromised
individuals. Therefore, workers involved in the composting process may be
exposed to potential health risks due to their exposure to A. fumigatus.
The term "Aspergillosis" has been used to describe illness attributed to
antigenic stimulation, colonization, or tissue invasion by A. species.
Aspergillosis is usually acquired by inhalation of airborne spores.
These spores (conidia) are small enough (2.5-3.Qium for A. fumigatus)
to reach alveoli or to gain entrance to paranasal sinuses.The disease
varies in severity from an incidental, saphrophytic relationship with
the host to a fulminating, fatal infection.
17
-------
The dose (number of cfu), portal of entry, and the immune status of the
host are thought to be major determinants in the course of the infection
caused by /\. species. Immunosuppressed patients are at a greater risk to
infection by A. fumigatus (24). Serious and often fatal invasive in-
fections caused by A. fumigatus have been reported in immunosuppressed
patients following kidney transplanation and in patients with leukemia or
lymphoma receiving chemotherapy. Individuals receiving antibodies or
adrenal cortical hormones have also been shown to have a higher incidence
of mycotic infections. Additional predisposing factors to aspergillosis
appear to be the presence of malignant or other debilitating disease,
leukopenia or granulocytopenia, other infections, pneumonitis and underlying
pulmonary disease (24).
Exposure of atopic individuals, who have a history of asthma, to spores
of /\. fumigatus may result in a disease known as allergic bronchopulmonary
asperillogis.Colonization of the bronchi in these patients with
Aspergillus results in episodic bronchial plugging which appears to lead to
areas of sacular bronchiectasis. These patients may produce sputum plugs
which often reveal fungal mycelia on microscopic examination. Laboratory
abnormalities in these patients include significant eosinophilia of blood
and sputum, marked elevation of total serum IgE, and serum precipitating
antibody to Aspergillus antigens. These patients usually have an immediate
type skin response to Aspergillus antigen. Persistent bronchopulmonary
aspergillosis may result in irreversible complications such as bronchiectasis
and pulmonary fibrosis.
The pathophysiology of allergic bronchopulmonciry aspergillosis is
thought to be mediated by a combination of both Type I and Type II immuno-
logic reactions. IgE-sensitized mast cells in the bronchi react to anti-
gens from Asp_ejxnJ_lus colonies growing in the bronchi. These mast cells
release histamine, slow-reacting substance of anaphylasix and eosinophilic
chemotactic factor leading to bronchospasm, increased permeability of the
bronchial mucosa, absorption of Aspergi11 us antigen into the circulation,
and pulmonary and peripheral blood eosinophilia. The absorbed Aspergillus
antigen can react with IgG, resulting in the formation of antigen-antibody
complexes and complement fixation. This in turn, may lead to chronic in-
flammation in the bronchi and peribronchial tissues with eventual
bronchial destruction, bronchiectasis, and pulmonary fibrosis.
Due to the wide distribution of A. species in the environment, ex-
posure to Aspergillus must be nearly universal, but disease, either in-
vasive or allergic, is uncommon. As indicated above, a complex interaction
between host factors and the challenge dose of A_. species spores appears to
determine the course of events which follow exposure to Aspergillus.
To date, no known adverse health effects have been detected among
compost workers due to exposure to thermophilic fungi and/or actinomycetes.
The Los Angeles County Sanitation District has been composting sewage
sludge for many years using the windrow method. The compost produced is
marketed by Kellogg Supply Co., Inc., for use in a wide variety of
applications. In the several decades of the operation of this system,
18
-------
there have been no reports of adverse health impacts (9), however, due to
the apparent transient nature of the work force the efficiency of the re-
porting system is probably low.
Composting by the forced aeration method has been carried out in
Beltsville since March 1973, and for shorter periods of time in other
locations. In the accumulated time of system operation and worker and user
exposure, there have thus far been no reported cases of disease resulting
from exposure to compost.
ELEMENTS OF COMPOST WORKERS STUDY
This investigation was designed to evaluate the health effects related
to exposure to nonviable substances and viable microorganisms present in
dust arising from the composting of sewage sludge. The clinical and
serologic evaluation of workers exposed to dust at the composting site in-
cluded: 1) comprehensive history and physical examination; 2) health
questionnaires; 3) illness .monitoring; 4) liver and kidney function pro-
files; 5) anterior nares and oropharyngeal swab cultures; 6) chest X-ray;
7) complete blood count with differential; 8) determination of antibodies
directed against A. fumigatus; 9) determination of antibodies directed
against 1ipopolysacchariaeTTCPS) present in compost samples; 10) skin
testing with A. fumigatus antigen; 11) pulmonary function tests; 12)
quantitation of total IgG, IgM, IgA, and IgE; and 13) determination of
C-reactive protein and levels of C3 and CHgQ. Health questionnaires, an-
terior nares and oropharyngeal swab cultures and sera for each of the
serologic tests were obtained five times during 1979, the first year of
the investigation. Environmental monitoring for viable particles was also
conducted several times during the first year. The scheduling of the above
elements of the study protocol is outlined in Table 1.
Population Groups Selected
Workers at sludge composting facilities in Camden, N.J., Beltsville,
M.D. and Washington, D.C. were selected because of their geographic
closeness and the willingness of their organizations to cooperate with the
study. Control workers were recruited either from sewage treatment
plants producing the sludge, if the job locations were judged to be
sufficiently distant from the compost operations, or from other nearby
sewage treatment plants. Compost treatment facilities in Camden, N.J.
and Washington, D.C. are operated by the Camden County Municipal Utilities
Authority and by the District of Columbia Department of Environmental
Services, respectively. In Camden the compost unit is located at the main
wastewater treatment plant and in Washington, D.C. at the Blue Plains Waste-
water Treatment Plant. Permission was not received from the U.S. De-
partment of Agriculture to recruit their compost-exposed employees at the
Beltsville Agricultural Research Center, but it was received from the Mary-
land Environmental Services which employs many of the workers involved
with the Beltsville compost-related activities. Nonexposed workers in the
Camden area were recruited from a small secondary wastewater treatment plant
in Camden, the Baldwin Run Plant. In the Washington, D.C.-Beltsville, MD
19
-------
TABLE 1. SCHEDULING OF ELEMENTS OF COMPOST WORKERS STUDY
Study element
Comprehensive history
Physical examination
Illness monitoring
Anterior nares and oropharyngeal
swabs for A. fumigatus
Chest X-ray
1
X
X
X
Month
5 T
X X
X X
(ONCE)
9
X
X
X
12
X
X
Complete blood count with
differential
Fungal serology (A, fumigatus,
A. carneus^ A. nTg"er and A.
f 1avusT*
Determination of antibody
directed against Lipopoly-
saccharide (IPS) prepared
from compost
Skin testing with A. fumigatus
antigen preparation** X
Pulmonary function testing X X XX
Immunochemical determination of
CRP, C3,-and immunoglobulins
(IgM, IgG, and IgA) x X X X
IgE determination
Environmental monitoring XXX
Renal and liver profiles X
X
X
X
X
x
*Aspergillin, Meridian Diagnostics, Cincinnati, Ohio.
**A. fumigatus allergic extract for scratch test (1-10 W-V)
Hollister-Stier Lab., Spokane, Washington.
20
-------
located about 10 miles down the Potomac River from the Blue Plains Plant.
The Beltsville compost facility treats a small portion of the sludge pro-
duced at the Blue Plains Plant. Another portion of the Blue Plains Plant
sludge is trenched elsewhere in Maryland. The Camden compost facility
treats sludge from the treatment plant at which it is located as well as
that from the smaller Baldwin Run Plant. The size and type of treatment
facilities from which these study participants were recruited are given in
Table 2. A total of 170 workers were recruited from these plants for the
study that is now being reported. About 100 additional low exposure workers
have been recruited from the Blue Plains Plant.
TABLE 2. SIZE AND TYPE OF TREATMENT FACILITIES FROM WHICH
PARTICIPANTS WERE RECRUITED
Location
-.-... wastewater
Size (MGD) Type
Compost Date
Size (dry tons/day) Started
Camden, NO
Main Plant 30 Primary
Baldwin Run . 4 Secondary
Beltsville, MD . None——
Washington, D.C.
(Blue Plains) 300 Secondary
P i scataway, MD 30
15
•-None-
15
75
5/78
3/73
2/79
•None-
Compost workers and control subjects in Philadelphia, PA (30) and Bangor
and Portland, ME (20), have also been recruited. Results from these
workers will be included in a future report.
METHODS
Clinical and Serological
The study was designed to evaluate the participants for objective and
subjective symptoms and signs possibly related to exposure to dust associated
with the composting operation. Particiants received physical examinations
once during the first year of the study. In addition, a detailed medical
and occupational history, which included reference to contact with sewage
sludge composting operations, or any other occupational dust and/or waste
exposure was obtained from each participant. This data base allowed
ranking of the participants in this investigation in terms of- length and
severity of exposure to the environmental dust and other factors associated
with composting. A health questionnaire, specifically designed to determine
the occurrence of hypersensitivity, respiratory, gastrointestinal and
mucocutaneous disorders associated with dust exposure, was obtained from
21
-------
the participants during the first, fifth, seventh, nine and twelfth months of
1979. IPS (endotoxin) was extracted from bulk compost samples from each
composting site by the method of Westphal (25). The serum from the study
population was tested for the presence of antibodies directed against each of
the IPS preparations by the enzyme-linked immunosorbent^assay (ELISA). Be-
cause of the difficulty in demonstrating infection with'members of
Aspergillus species, four parameters were employed to detect infection with
these fungi: 1) determination of precipitating antibody directed against A.
fumigatus, A. carneus, A. niger and A_. flavus in the serum of the study pop-
ulation, 2) determination of skin test reactivity to an extract of A.
fumigatus, 3) semi-quantitative sputum cultures for A. fumigatus,an? 4)
evidence of transient or migratory infiltrates on chest X-ray.
Precipitating antibody to A. fumigatus, A_. flavus, A. carneus and A_.
niger (Aspergillin, Meridian Diagnostics, Cincinnati, Ohio) in the serum of
the study population was measured semi-quantitatively by titration of anti-
body using the counter immunoelectrophoretic technique (26). Antigens of A.
fumigatus used in the determination of specific IgG antibody by the ELISA
were prepared by the method of Coleman and Kaufman (28) and further purified
by gel filtration on Sephadex G-200. The first peak obtained by gel
filtration of the antigen preparations were be used for determination of
precipitins in the serum samples tested. Sera positive for /\. fumigatus were
kindly provided by Dr. 0. W. Rippon, University of Chicago Hospital, Chicago,
IL, and were used as positive controls for precipitin determinations. Serum
samples for fungal precipitin determination were obtained from the study popu-
lation during the first, fifth, seventh, ninth and twelfth months of the first
year of the investigation.
Skin.reactivity to Aspergullus antigen was determined by scratch test,
using Aspergillus antigen (Hoilisten-Stein Corp., Spokane, WA) in 1:10
weight/volume concentration. The skin test was performed during the twelfth
month of the study. Determination of total IgE titers were performed by the
ELISA method using sera collected during December 1979.
Anterior nares and oropharyngeal swabs were obtained from the study pop-
ulation during the fifth, seventh, ninth, and twelfth months of the first year
of the investigation and cultured for A. fumigatus. Sputum specimens were ob-
tained during the first month but were discontinued because of the difficulty
in obtaining adequate samples. Due to the widespread distribution of A.
fumigatus, semi-quantitative determination of A. fumigatus in the samples
obtained from the study and control groups was performed. Quantitation
of the number of A. fumigatus in the samples may provide an index of
the degree of colonization between the groups participating in the
study.
Transient or migratory pulmonary infiltrates are frequently observed
in individuals with hypersensitive lung diseases. A chest X-ray was
obtained on participants once during the first twelve-month period
and any infiltrative process or other lung pathology was recorded. In
the future, prior chest X-rays of the study participants will be re-
viewed, if possible, and any positive findings will be correlated
22
-------
with the clinical diagnosis made at the time the X-ray was taken and with
periods of exposure to the composting operation.
Chronic exposure to microbial antigens present in the dust arising from
the composting of sewage sludge may induce a local and/or systemic in-
flammatory response as well as an elevation of one or more classes of circu-
lating immunoglobulins. C-reactive protein and C3, serum proteins which are
often elevated during inflammatory processes, were quantitated by radial
immunodiffusion (28). Hemolytic complement was also assayed (26)since it may
be reduced in immunological phenomena involving antigen-antibody complexes.
The concentration of total IgG, IgM, and IgA was quantitatively determined in
the sera of the study population by radial immunodiffusion (28).
The study population was also evaluated for evidence of hypersensitive
pulmonary disease or other lung diseases which may be associated with pro-
longed exposure to the dust arising from the composting operation. Each
participant in the proposed investigation underwent pulmonary function
testing. When possible, pre- and post-shift testing was performed. It was
felt that pulmonary function testing in this manner would detect bronchial
sensitivity associated with exposure to dusts in the work environment.
Environmental Monitoring
Viable particle collection from air samples was performed using the
six-stage Andersen cascade impactor designed for that purpose. Air was drawn
through the samplers by 12 volt D.C. pumps, which were powered by 12 volt
motorcycle batteries. The pumps, attached to an assembled sampler loaded with
petri dishes containing agar, were calibrated against a dry gas meter to pull
one cubic-foot of air per minute (CFM), as recommended by the manufacturer.
The samplers were mounted on tripods such that samples were collected at a
height of five feetj which approximate the breathing zone of the average
worker. Sampling times were adjusted in an attempt to obtain colony numbers
which were optimal for counting. The volume of air sampled was calculated
from the sampling time and the flow rate of the pumps. Total colony counts for
bacterial and fungal plates were obtained by direct colony count, unless
excessive colony numbers precluded this procedure. If colony numbers pre-
cluded counting all colonies, the colonies on 1/4 or 1/2 the plate were
counted and the count was multiplied by the appropriate number to obtain a
'total plate count.
Bacterial samples were collected on Trypticase soy agar (TSA) manu-
factured by Baltimore Biological Laboratories (BBL), Cockeysville, MD. The
medium was prepared according to the manufacturers instructions, autoclaved at
121°C for 15 minutes, cooled to 45-50°C and dispensed into petri dishes.
Prior to use, TSA plates were incubated at 36°C for 24-48 hours to detect
contamination and minimize surface moisture due to condensation.
Fungal samples were collected on modified Czapek-Dox medium. The medium
was prepared by adding 35 grams of Czapek- Dox broth (Difco Laboratories,
Detroit, MI), 20 grams agar (Difco), and 15 grams Oxgall (Matheson, Coleman
and Bell, Cincinnati, OH) to 1000 milliliters distilled water, adjusting the
23
-------
pH to 7.3 and autoclaving at 121°C for 15 minutes. Prior to dispensing, the
medium was cooled to 45-50°C and 50/ig/ml Streptomycin, 50>ug/ml Chloram-
phenicol and 20 units/ml Penicillin were added to retard bacterial growth.
These plates were incubated at room temperature or occasionally at 36°C to
minimize moisture.
Aerosol samples obtained on ISA plates were shipped directly (within 24
hours) to the Research Surgical Bacteriology Laboratory at the University of
Cincinnati, where they were incubated at 36°C for 24 hours. At the end of the
incubation period, total bacterial counts were made from the TSA plates which
were then replicate plated onto three different types of media: KF strep-
tococcal agar (BBL), M - FC agar (BBL), and M-Endo agar (LES) (BBL) (30).
KF streptococcal agar is a selective medium which supports the growth of
fecal streptococci (Lancefield's serological groups D and Q) while inhibiting
the growth of most gram-negative and other gram-positive bacteria. The
presence of pink/red colonies after 48 hours at 36°C is indicative of fecal
streptococci. All growth on KF streptococcal agar was re-replicated to
Bile-Esculin agar (Difco) and tested for catalase activity. Gram-positive
cocci which grew as brown/black colonies on Bile-Esculin agar, and which
failed to decompose a three percent solution of hydrogen peroxide, were con-
sidered to be confirmed fecal streptococci.
The purpose of replication onto M-FC agar was to determine the presence
or absence of fecal coliforms. These plates were incubated at 44.5°C for 24
hours and the blue to blue green colonies were transferred to a Lauryl sulfate
broth (BBL) which was incubated 24-48 hours at 37°C and to EC broth (BBL)
which is incubated for 24 hours at 44.5°C. Growth with gas in these media
confirmed ..the presence of fecal coliforms.
In order to determine total coliform counts, the TSA plates were rep-
licated onto M-Endo agar (BBL) and incubated at 37°C for 24 hours. The
colonies with a green sheen were picked to Lauryl sulfate broth and Brillant
green bile broth (BBL). These media were incubated at 37°C for 48 hours, + 3
hours. Growth with gas confirmed the presence of coliforms. ~
Fungal plates, upon reaching the laboratory, were incubated at 42.5 to
44.5°C for 48 hours and total plate counts were performed. The microorganisms
were separated into A. fumigatus and "other thermophilic microorganisms" on
the basis of microscopic examination and colony color and morphology.
RESULTS
In This report only a preliminary analysis of a portion of the data from
participants at Camden, Beltsville, Washington, D.C. (Blue Plains) and
Piscataway is available. Data from workers in Philadelphia, PA, Bangor and
Portland, ME and from the additional lower exposed Washington, D.C. volunteers
will be presented in a future report.
Populations Recruited
A summary of the 173 workers recruited from Camden, N.J., Beltsville, MD,
24
-------
Washington, D.C. (Blue Plains) and Piscataway, MD is presented in Tables 3, 4,
5 and 6, respectively. Initial recruitment began in late January, 1979 in
Camden, N.J. and in March for Beltsville, MD, Washington, D.C. and
Piscataway, MD. The participants from each of these areas have been assigned
compost exposure categories: high, intermediate, or control. Assignment to a
particular category was based on interviews and direct observations. Workers
who are directly involved in compost operations as their major activity were
assigned to the high exposure category. The intermediate category applies to
workers either routinely working within about 100 meters of a compost operation
or who are sometimes involved with composting operations but not as the
major part of their work. These categories are relative ones and are area
specific; that is an "intermediate exposure" workers at the Camden compost
plant would not necessarily have the same exposure as an "intermediate
exposure" worker at Blue Plains.
In Camden the intermediate exposure category was used for workers at
the Main Plant who were not involved in the compost operation. Control
workers in Camden were from the Baldwin Run Plant. There were 21 Camden
participants in the high exposure category, 22 in the intermediate and 14
in the low exposure group. At Beltsville, MD there were 10 people in
the high exposure group working at the compost site 'and-there were eight in the
intermediate group who primari-ly worked with compost samples at a research
laboratory some distance from the compost and on occasion visited the
compost site for sample collection and other purposes. Included in the
high exposure group was one person whose job was based in a trailer
located at the compost site. In Washington, D.C. 14 compost workers have
been recruited in the high exposure group. An intermediate exposure
group consisted of 27 workers whose jobs occasionally required them to
repair equipment in the Blue Plains compost pit and on occasion at the
BeltsYiHe compost site. A control group of 11 workers was recruited ,
who report to a pumping station several nil!es from the Blue Plains Plant but
spend a portion of their time at the Blue Plains Plant maintaining the
pumps in the raw sewage inlet station and in the air blower building,
both of which were several hundred meters from the compost pit. The 40
participants recruited at the Piscataway, MD plant were engaged in various
operational and laboratortory functions. All have been assigned to the control
category since no composting operations are involved at their plant. The median
age for the Camden groups ranged from 28 to 37 years, at Beltsville, from
27 to 47, at Blue Plains from 31 to 42, and at Piscataway the median age
of the one group was 30 years. The gradations of exposure at Camden,
Beltsville and Blue Plains allowed for comparison of results between
workers at these locations. In addition, Blue Plains workers were com-
pared to the control group at nearby Piscataway, MD.
Air Sampling
During the period May-December, 1979 viable particle air sampling was
conducted at several locations at each of the treatment facilities involved
in the study. Results are summarized for /\. fumigatus and "Other Thermophilic
Microorganisms" in Table 7, for Fecal Streptococci and Fecal Coliforms in
Table 8, and for Total Coliform and Total Bacteria in Table 9. At the Blue
25
-------
TABLE 3. CAMDEN WORKERS: (AGE. RACE. SEX AND EXPOSURE)
Exp.
High
Int.
Cont.
No.
19(1)
19
7
Whites
Med.
28
26
27
Range
19-61
19-70
23-61
Non-whites
No.
2
9
7
Med.
55
48
43
Range
48-61
25-68
22-64
Total
No.
21(1)
28
14
Med.
29
28
37
•
Range
19-61
19-70
22-64
Number of which are females appear in parentheses.
TABLE 4. BELTSVILLE WORKERS: (AGE, RACE, SEX AND EXPOSURE)
EXD.
High
Int.
Cont.
No.
9U)
8(2)
0
Whites
Med.
50
27
Range
29-59
25-65
No.
1
0
0
Non-whites
Med.
43
Range
,_ Total
No.
10(1)
8(2)
0
Med.
47
27
Range
29-59
25-65
Number of which are females appear in parentheses.
TABLE 5. BLUE PLAINS WORKERS: (AGE. RACE. SEX AND EXPOSURE)
Exp.
High
Int.
Cont.
No.
1
5
2
Whites
Med.
32
35
36
Range
26-45
29-42
Non -whites
No.
13(1)
22(6)
9
Med.
37
31
42
Range
20-55
23-49
23-51
Total
No.
14(1)
27(6)
11
Med.
37
31
42
Ranae
20-55
23-49
23-51 __
TABLE 6. PISCATAWAY WORKERS:
Exp.
High
Int.
Cont.
No.
0
0
18(1)
Whites
Med.
28
Range
22-63
Non-whites
No.
0
0
22(6)
Med.
31
Range
25-47
Total
No.
0
0
40(7)
Med.
30
Ranqe
22-63 '•
Plains and Main Camden plants results were summarized for samples collected in
the vicinity of the compost area and at locations more distant from the com-
post areas. Because of the much larger size of the Blue Plains Plant, "Other
Areas" for it were at least 150 meters from the compost area while they were
only at least 50 meters away at the Camden plant. (At the Camden plant, all
areas of the plant were within 150 meters from the compost area.) Samples
were collected at each plant on from three to six days except for Beltsville
where samples were collected on only one day. Only colonies from plates 3-6
(respirable size range) from the Andersen samplers were included in Tables
26
-------
ro
7. SUMMARY OF 1979 AIR MONITORING FOR RESPIRABLE CONCENTRATIONS OF A. FUMIGATUJ
AND OTHER THERMOPHILIC MICROORGANISMS {COLONY FORMING UNITS PER CUBIC MTTE^
A. fumiqatus
Area No.
Metro. Washington, D.C.
Beltsville, Compost
Blue Plains, Compost
Blue Plains, Other Areas*
Piscataway Plant
Camden, N.J.
Main Plant, Compost
Main Plant, Other Areas**
Baldwin Run Plant
samples
4
25
14
15
13
6
11
Range
48-131
0-475
0-21
0-18
0-2940
0-126
0-52
Average
81
52
3
3
918
24
14
Concentrations
CFU/M-3)
Other thermophilic microorganisms
Range
0-34
0-12,000
0-246
0-5
0-580
0-7
0-7
Average
17
1060
73
<1
146
2
1
*At least 150 meters from compost area.
**At least 50 meters from compost area.
-------
PO
CO
TABLE 8. SUMMARY OF 1979 AIR MONITORING FOR RESPIRABLE CONCENTRATIONS OF FECAL
STREPTOCOCCI AND FECAL COLIFORMS (COLONY FORMING UNITS PER CUBIC METER, CFU/M3)
Area
Metro. Washington, D.C.
Beltsville, Compost
Blue Plains, Compost
Blue Plains
Piscataway
;amden, N.J.
Main Plant,
Main Plant,
Baldwin Run
, Other Areas*
Compost
Other Areas**
Plant
No. samples
4
4
0
6
8
2
6
Concentrations
Fecal streptococci Fecal conforms
ange
0-23
11-79
0-6
0-482
0-5
0-25
Average
6
48
1
85
3
8
Kange
0
n
0-3
o
0-5
0-21
Avarage
0
1
3
5
*At least 150 meters from compost area.
**At least 50 meters from compost area.
-------
V£>
TABLE 9 SUMMARY OF 1979 AIR MONITORING FOR RESPIRABLE CONCENTRATIONS OF COLIFORMS AND
TOTAL BACTERIA (COLONY FORMING UNITS PER CUBIC METER.
Concentrations
Total conforms Total bacteria
Area
.Metro. Washington, O.C.
Beltsville, Compost
Blue Plains, Compost
Blue Plains, Other Areas*
Piscataway, Plant
Camden, N.J.
Main Plant, Compost
Main Plant, Other Areas**
Baldwin Run Plant
No. samples
4
4
0
6
8
2
6
Range
0
0
0-27
0-56
0-10
0-35
Average
0
0
.12
8
5
10
Range
5, 550- 16 i 800
8,070-18, 100
564-1330
590-5170
443-524
196-3380
Average
10,700
14,600
815
2720
484
1460
*At least 150 meters from compost area.
**At least 50 meters from compost area.
-------
7-9. Concentrations of A. fumigatus varied considerably and on average were
higher in compost areas Than in noncompost areas. Concentrations at areas at
least 150 meters from the compost pit at the Blue Plains Plant were about the
same as at the Piscataway Plant where no composting occurs. At the Main
Camden Plant, A. fumigatus levels in areas at least 50 meters from the compost
pit were on average much lower than concentrations in the compost area, but
were on occasion higher than those at the Baldwin Run Plant. Concentrations
of A. fumigatus at Camden were generally higher than those at Beltsville and
Blue Plains. These differences may be due in part to differences in compost
activities underway at the time of the sampling or to differences in pre-
cipitation levels prior to sampling. Two days prior to one of the Blue Plains
sampling days, about four inches of rainfall occurred in the vicinity of the
plant as tropical storm David passed through the D.C. area. About six percent
of all colonies of A. fumigatus were found on plate 6 and were thus thought to
be small enough to reach the alveoli. This compares to less than one percent
found to be in that size range by Mi liner et al. (20). About 80 percent were
found to be in the respirable size range (plates 3-6) which is practically the
same as that reported by Mi liner et al. (20), 79 percent.
In Camden the two treatment plants are clearly distinguisable from each
other with respect to A. fumigatus levels. At the main Camden plant A.
fumigatus levels were considerably higher at compost areas than elsewhere in
the plant where samples were collected. Similarly, A. fumigatus concen-
trations at the Piscataway Plant and at areas of the Blue Plains Plant remote
from the compost pit were clearly lower than levels reached at the Beltsville
and Blue Plains composting sites.
Concentrations of other thermophilic microorganisms varied widely but
were generally higher at the compost sites.
Fecal streptococci were detected at each of the areas where they were
measured and were generally higher at the compost areas (Table 8). Fecal
coliforms were infrequently detected (Table 8). Coliforms (Table 9) were not
detected in samples collected at Beltsville and Blue Plains. Concentrations
were about the same at Piscataway as they were at the Camden sampling
locations (averages ranged from 5 to 12 cfu/M3). Concentrations of total
bacteria ranged somewhat higher at compost sites than at other sites but were
above 10^ cfu/M3 in all samples.
A. fumigatus in Study Participants
Culturing for of A. fumigatus colonies was performed on sputum specimens
or from swab cultures of the anterior nares and oropharynx obtained from
study participants. Since not all participants were available for testing
during each of the five visits, between one and five determinations were made
for each individual. A summary of the highest value for each individual,
according to location and exposure-category, is presented in Table 10. The
number of colonies observed are expressed as either zero or greater than or
equal to one. Statistical analyses by the chi-square test, or Fisher's exact
test ^1J when appropriate, were performed between results for the warion^ exoosur
groups and combinations of exposure groups in each city and betweln groups for
30
-------
co
TABLE 10 DISTRIBUTIONS AND ANALYSES OF STUDY PARTICIPANTS ACCORDING TO THE HIGHEST VALUE
OF ASPERGILLUS FUMIGATUS (AF) COLONIES OBSERVED ON CULTURES TAKEN DURING 1979
Exposure Categories
1 2 3
Camden
AF^l 16
AF r 0 5
Blue Plains
AF >1 8
AF* 0 5
Blue Plains &
Piscataway
AF s«l 8
AF= 0 5
Beltsvllle
AFstl 7
AF =0 3
All Sites Combined
AF * 1 31
AF=-0 13
5 1
20 12
7 0
20 11
7 2
20 48
0
8
12 3 •'•"
48 60
Exposures
Compared
1:2
1:3
1:2,3
2:3
1:2
1:3
1:2,3
2:3
1:2
1:3
1:2,3
2:3
1:2
ill
1:2,3
2:3
Testing gCHypql-hesis of Independence
\P values J
X2 Test Fisher's Exact Test
<.0005
<.0005
<.05
X..005*
<..05
<..005*
<".0005
«..01
14
-^ .002
.070
^..0001
— .007
^w.004
*Small cell expected value renders the validity of the Xz test "borderline."
-------
all areas combined. The null hypothesis tested is that the probability of a
positive count is independent of exposure cagegory. Statistically significant
differences were found between many of the groups with those in exposure
category one having more positive determinations than either those in category
two or category three. Workers in category 2 had more positive values than
those in group three and the results were statistically different for Blue
Plains and Piscataway combined and for all sites combined.
The differences among the groups in A. fumigatus colonies verified the
overall validity of the exposure categories. Even though viable A. fumiqatus
(AFJ spores are ubiquitous in the environment, the increase, above" ambient
levels in their concentration at or near compost sites was apparently large
enough to result in a higher degree of incidence among workers with compost
exposure than in those without. Combining all sites, the percentages of study
participants exhibiting at least one. positive value for AF colonies, distri-
buted by exposure category, were category one (high exposure 70%: category
two (intermediate exposure) 20%; category three (control) 5%.
Liver Function and Total Immunoglobulin Determination
Preliminary analyses of the highest results obtained for each participant
on tests for the enzymes serum glutamic oxaloacetic transaminase (SCOT), serum
glutamic pyruvic transaminase (SGPT), measures of liver function, and levels
nLnncaLl9G' lf^nA !§M' did not ^^ *n* differences among exposure
groups- at any of the sites. Combining data by exposure group for all sites
again did not reveal any significant difference among exposure groups.
A more detailed examination of the data may reveal differences In mean values
tor the various exposure groups, however.
Delayed Hypersensitivity to Asoergillus Antigens
n nhar? A'Per^111us ant19en was determined on 109 participants
one intPrmJ^ 0nly/1Ve, persons tested Positive, three compost workers,
were
C-Reactive Protein (CRP)
Derormeonwn n °Vhe highest CRP value for each individual was
statist callv siSniJiJ.nt no;r?act™e results revealed only one borderline
of n ne comiXst SrklrJitT?!* ?.= :°53) by the Fisher's e*act test: Four
noneTf »^£4»^ ">r CRP compared to
Eosinophil Count
the piciS K££ "*&*»*«* *osinophil «unt for each of
the various sites, separately or combined **'"* ^ eXP°SUre 9I"°UPS
32
-------
Chest X-Ray Findings
X-rays were obtained from 74 participants and were reviewed by a radio-
logist at the Un.iversity of Cincinnati. Although a total of 17 abnormal
findings were observed, only two were suggestive 'of occupational exposure or
smoking, one a compost worker and one a control.
Antibody to A. funrigatus
Specific IgG directed towards A. fumigatus, as determined by the ELISA,
was compared by exposure group for each site separately. Statistical analyses
were performed by the Mann-Whitney one-tailed ranking test. At no site did
the compost workers exhibit statistically higher values.
Antibody to Lipopolysaccharide (IPS)
IPS was prepared from compost samples obtained from the Camden site, and
from the Beltsvilie-Blue Plains sites combined, by the Westphal method and
tested against workers sera to determine possible responses to IPS present in
the compost material. Specific IgG directed against the IPS was determined by
the ELISA. Statistical analysis was by the Mann-Whitney one-tailed ranking
test. Antibody titers in the Camden compost workers (group one) directed
against IPS prepared from compost obtained from Camden were not higher than in
the control workers (group three). However, in Beltsvilie, the compost
workers had higher antibody titers against IPS prepared from compost obtained
from the Blue Plains-Beltsville compost sites, than the intermediate group
(p = .04). At Blue Plains, antibody titers in the compost workers were higher
than in the intermediate group (p = .025); when group three (control) workers
at Piscataway and Blue Plains were included the significance was still
p = .02.
Health Effects Associated with Composting
A review of the medical histories, physical examinations and chest X-rays
of compost workers participating in the study has thus far revealed one worker
with several health problems which may be associated with exposure to micro-
bial pathogens and dust present at the composting site. Approximately one
year after starting employment at a compost site, the worker developed
chronic otitis media of the right ear, with sclerosis of the right mastoid and
erosion of the posterior wall of the right external canal demonstrated on
X-rays of the mastoid. Over a period of several months the infection grad-
ually progressed, with erosion of 70% of the right tympanic membrane, despite
topical antibiotics, local debridement, and a right tympanomastoidectomy.
Aspergillus niger, a fungus which has been reported present in compost, was
the only pathogen isolated from the purulent discharge present in the right
ear. This worker also has small irregular shadows in both lungs suggestive of
parenchymal disease. Since this individual states that he has never smoked,
the pulmonary abnormalities observed on the chest X-ray may be job related.
Except for evidence of the right chronic otitis, the physical examination of
this worker was normal, as were his pulmonary function studies on three
33
-------
different occasions. The worker also complained of symptoms suggestive of
chronic sinusitis, however, the relation of the onset of these symptoms and
his exposure to composting could not be definitely determined.
DISCUSSION AND CONCLUSION
Workers at sludge compost sites are exposed to a variety of fungal and
bacterial pathogens, as well as toxins of microbial origin, in their work
environment. No information is currently available regarding the potential
long-term effects of this exposure on the health of these workers. This re-
port has reviewed current literature related to host responses to micro-
organisms and microbial toxins known to be present at compost sites and has
presented preliminary results of a study designed to evaluate potential health
effects of exposure to microbial factors present at compost sites.
Results of the environmental monitoring demonstrated that workers at each
of the compost sites studies were exposed to markedly elevated numbers of A.
fumigatus spores. This observation was supported by the high frequency of
isolation of A. fumigatus from oropharyngeal and anterior nares cultures ob-
tained from the compost workers when compared to the other two exposure
groups. However, the presence of A. fumigatus in the anterior nares and
oropharyngeal cultures did not correlate with antibody response directed
against A. fumigatus in those workers with positive cultures or among the_
three exposure groups. One worker in the study, employed at a compost site,
was found to have a local infection involving his right ear caused by A. nl2££
and chest X-ray changes compatible with an occupationally-related disorder.
It has not been possible to document whether the ear infection or the chest
X-ray findings were directly related to exposure to microbiolic agents present
at the compost site.
Due to the large numbers of gram-negative bacteria in the sewage sludge
which is composted, it was assumed that workers at compost sites would be
exposed to relatively large amounts of IPS {endotoxin) in the dust at the
compost site. The only means by which this exposure could be documented was
by the viable counts of gram-negative microorganism (Fecal Coliforms) col-
lected during environmental monitoring. These levels may be artifically low
since a major component of the LPS in dust may be comprised of nonviable
bacteria. The LPS (endotoxin) used for determination of antibody titers in
the study participants was prepared from compost obtained from the Camden and
Beltsville-Blue Plains compost sites. Levels of specific IgG antibody di-
rected against LPS prepared from Beltsville-Blue Plains compost were signif-
icantly higher in the Beltsville and Blue Plains compost workers than in the
nonexposed controls in the Washington, D.C. area. No difference in specific
IgG antibody levels directed against LPS prepared from Camden compost was
observed in exposed and unexposed workers in the Camden, NO area. The immune
response observed in the Blue Plains-Beltsville compost workers to challenge
with LPS by the respiratory route was similar to that reported by Rylander et
al. for workers in cotton mills exposed to gram-negative bacteria in cotton
dust and rats challenged with purified E. coli 025:66 LPS (14 15)
34
-------
Preliminary analysis of other parameters studied, such as C-reactive
protein, total immunoglobulin (IgG, IgM, IgA, IgE) levels, liver function
tests, skin test reactivity to A. fumigatus, chest X-rays and absolute eo-
sinophilcounts, did not reveal significant differences among the three ex-
posure groups. When the available data is subjected to a more complete
analysis, incorporating such factors as age, race, precise length of exposure,
the preliminary findings may require revision. In addition, an extended
period of observation may reveal abnormalities which are related to the length
of exposure. The preliminary data on antibody response of the compost workers
at the Beltsville-Blue Plains sites to IPS suggest that it may be advisable to
take reasonable precautions to reduce exposure such as the use of respirators
by compost workers and periodic water spraying of the compost sites to reduce
dust.
35
-------
SECTION 5
AN ENVIRONMENTAL HEALTH SURVEY OF DRINKING WATER CONTAMINATION BY
LEACHATE FROM A PESTICIDE WASTE DUMP - HARDEMAN -COUNTY, TENNESSEE
Toxic waste dumps are a major source of environmental pollution in
many areas of the country and are being recognized with increased frequency.
In Hardeman County, Tennessee, wastes from a Memphis, Tennessee, pesticide
manufacturer have contaminated the groundwater aquifer serving about 60
nearby residents. During the years of operation, 1964-1972, an estimated
300,000- 500,000 barrels of liquid and solid waste were buried in shallow
trenches dug into the ridges at the approximately 200 acre dump site.
The dump was closed by state authorities in 1972 because contamination
had been detected in test wells close to the burial areas (32,33). The
analyses performed at that time did not reveal any contamination in the
three private water wells closest to the dump which were being monitored.
However, during the fall of 1977, nearby residents became alarmed upon
detection of unusual and unpleasant tastes and odors in their well water.
Within a short time, the residents reported an unusually high number of
illness symptoms including skin and eye irritation, weakness in the upper
and lower extremities, upper respiratory infection, shortness of breath
and severe gastrointestinal symptoms including nausea, diarrhea and abdominal
cramping. In general, these symptoms appeared to subside once ingestion
of the contaminated water was stopped. In an effort to determine the extent
and possible etiology of the signs and symptoms of illness, a limited health
screening study was conducted by investigators from the University of
Cincinnati in November 1978. The results from this study showed biochemical
changes suggestive of liver dysfunction in some of the residents whose
water was contaminated. A second more extensive evaluation was carried out
in January 1979. A 1978 investigation by the U.S. Geological Survey con-
firmed the residents' observations that contamination had occurred (34).
BACKGROUND
The Hardeman County toxic waste dump first came to the attention of
the investigators during the course of a seroepidemiologic research study
of wastewater treatment plant personnel in Memphis, Tennessee, which is
located about 60 miles from the dump site. One of the two treatment plants
involved in this study served an area which included the pesticide manu-
facturer whose waste had formerly been deposited in the Hardeman County
toxic waste dump. Chemical wastes from this manufacturer had apparently
been routinely discharged into the Memphis sewers and flowed into the Memphis
North wastewater treatment plant beginning with its operation in August
1977. A portion of these wastes became airborne at the treatment plant
36
-------
because of the aeration process utilized and because of the open basins
that are generally used in such facilities. Workers at the North Plant
began complaining of eye irritation and respiratory distress in the spring
of 1978 during periods of time when chemical odors at the treatment plant
were unusually Intense. In conjunction with the ongoing study at the treat-
ment plant, a urine screening study of the workers was conducted in May and
June 1978, and air samples were collected at various locations in the treat-
ment plant. Hexachlorocyclopentadiene (HEX) and hexachlorobicycloheptadiene
(HEX-BCH), two compounds produced by the pesticide manufacturer, were
detected in air, wastewater and urine specimens obtained at that time. In
the June testing workers at the Memphis North Treatment Plant were found to
have an increased concentration of the HEX-BCH compound in comparison to
urine specimens collected early and late in the work shift (35). Because
the same chemical company was involved in the Memphis sewage treatment
plant situation and in the disposal of toxic waste at the landfill, an
effort was made to learn whether any exposure or ill health effects had
been detected in the community whose well water was contaminated.
The U.S. Environmental Protection Agency and the State of Tennessee,
by the latie part of 197 •'d detected more than one dozen chlorinated
organic compounds in * of individuals near the dump site (36).
Many of these chemic * were on the National Priority Pollutant
Consent Decree List -achloride was the most abundant con-
taminant detected. *rumental in bringing about the
health study whir First of all, on investigation
of this problem "1th study had been conducted
nor was one pi the users of this con-
taminated wat osticide compounds in
the sewage > '*heir symptomatology,
it was fel4
-------
TABLE 11. CONTAMINANTS DETECTED a IN PRIVATE WELLS IN TOONE-TEAGUE
AREA OF HARDEMAN COUNTY. TENNESSEE
Compound
Highest level detected '(ppb)
Eenzeneb
'" k p
.Carbon Tetrachloride0 (CC14)
Chlordene
Chlorobenzene.b
Chloroformb (CHC13)
Hexachlorobutadiene-b
Hexachloroethaneb
Methylene Chloride b
Napthaleneb
Tetrachloroethyleneb
Toluene'*
Xylenes .
15
• ' '
18,700
0.81
41
1,890
.2.53
4.6
2;2
160
$.7
2,405
52
1.6
aAdapted from U.S. Environmental Protection Agency, Region IV, March 9,
1979, Summary of USEPA and State of Tennessee Chemical Analyses (36).
bOn National Priority Pollutants Consent Decree List (37).
trations at which they were found. These results were compared to a control
group from Memphis which had been collected two months earlier.* The
location of the affected wells in relation to the dump site is shown in
Figure 1. ,
The blood specimens that were drawn were centrifuged and the serum was
frozen and shipped to Cincinnati;. for performance of routine liver function
determinations including serum garama glutamic transaminase (SGGT), serum
glutamic oxalic transaminase (SGOT), and serum glutamic pyruvic transamiase
(SGPT), alkaline phosphatase and, total bilirubin. These tests were per-
formed in the Clinical Laboratory of Cincinnati General Hospital. The
Hardeman County specimens were analyzed in November and the Memphis specimens
in September and January. The urine specimens and water samples were
*The November 1978 results were later compared to those from a local control
group obtained in January 1979.
38
-------
Fiaure 1 Location of contaminated private wells near
Hardeman County toxic waste dump (designated
by X).
39
-------
frozen immediately after collection and an aliquot of each was saturated
with sodium chloride and extracted with equal volume of petroleum ether.
The petroleum ether extracts were stored in a freezer until time of analyst^-
The air samples were collected using glass sampling tubes containing 150
mg of preextracted chromosorb 102 and desorbed with petroleum ether. AH o1
these specimens were analyzed by electron capture gas chromatography using
glass columns, packed with OV-10 or OV-17-QF1, at 17S°C and were performed
at the laboratories of the Department of Environmental Health at the
University of Cincinnati, College of Medicine, Cincinnati, Ohio.
The statistical analyses of the liver function tests were performed on
the controls using age, sex, and a factor for maturity level as variables,
covariant adjustments from this analysis were then used in analysis of
variance for the entire group of data.
When the preliminary results from the initial study were analyzed, it was
apparent that there were serum enzyme levels suggestive of liver dysfunctio
present in serum of those consuming the contaminated water. As a result, a
decision to expand the November 1978 study was made and this was conducteo
in January of 1979. The follow-up study consisted of a more detailed ques-
tionnaire including past medical history, alcohol intake, drug c0"5^^^^
history of hepatitis and other liver abnormalities, as well as general nea
survey questions. In addition to this, a clinical examination was PeJ"l.onJ
to assess blood pressure, liver size and other physical parameters. Blooa
was obtained while fasting for determination of serum bile acids, a liver
profile, a renal profile and the hepatitis serology. The hepatitis t^st^
were included to evaluate whether any differences in liver enzyme tests we
related to previous infection. Postprandial specimens were collected two
hours after a breakfast consisting of eggs, bacon or sausage, juice, toast
and coffee or milk for analyses for serum and urine bile acids. The stu°{
was conducted at the Chickasaw State Park, located about 10 miles from the
dump site. The study group was expanded to include not only area residents
whose wells were contaminated but a number of other people from local civj\i
organizations and church groups were invited to participate as controls who
lived in the same geographic area as those whose water supplies were con-
taminated. There was also another group which included those who were fre-
quent visitors to the households of those persons with contaminated water
or who were close neighbors whose water was not significantly contaminated
and were considered to represent an intermediate exposure population. A map
showing the location of the residences of the controls recruited for the
January 1979 study and there relation to the toxic waste dump is shown in
Figure 2. Persons with abnormal liver function tests in January 1979 had
repeat determinations performed on serum collected in March 1979.
Participants who had occupational exposure to pesticides, who had con-
sumed alcohol 24 hours prior to the collection of the blood specimens or WHO
had a history of a liver disorder were deleted from the analysis of liver
and renal profiles and serum and urine bile acids. The dependent variables
were then tested for normality and transformations were made when necessary-
40
-------
Figure 2. Location of homes of January 1979 control group in relation to dump site
(designated by letter C).
-------
Analysis of covariance was used to assess the effects of age, sex, monthly
alcohol consumption and their interactions on the dependent variables of the
controls. Adjustments were then made for those covariables which were found
to be significant. One-way analysis of variance was used to test whether
there were significant differences between the exposed, intermediate-exposed,
and control groups in the January 1979 study. When all three groups were
tested simultaneously, analysis of variance with 2 degrees of freedom was
used. Since this analysis involved multiple comparisons, the significance
level was set at 0.02. The exposed group from the November 1978 study was
also compared to the control group from the January 1979 study.
Representative air and water samples were collected from residences of
the exposed, intermediate-exposed and control groups. In some cases, indoor
air samples were collected in bathrooms with either the bathtub or the
shower tap running.
In order to test whether the 30 people in the study group who were
tested in both November 1978 and January 1979 showed a significant change in
their hepatic profile, the differences between the results of the two periods
were analyzed. These differences were found to be normally distributed
except for S6GT which required a log transformation of its substraction from
a constant. A t-test was then performed on the differences.
Results - November 1978
Water Use--
During the period from May to December 1978 a trailer-mounted water
tank served as a temporary supply for kitchen and drinking uses. Periodi-
cally the tank was refilled in a nearby community. A second temporary supply
piped from an underground holding tank to a number of the residences was
installed in December 1978. A permanent water supply from the village of
Toone was connected to most of the residents in 1979. On the day of the
collection of blood and urine from Hardeman County residents, many of the
people in the affected area had received certified letters from the U. S.
Environmental Protection Agency (Region IV) advising them not to use the
water for any purposes because of high amounts of carbon tetrachloride
found. Some of the residents had stopped using the water for potable uses
(drinking and cooking) beginning as early as the fall of 1977. A large
group stopped such use in May 1978 after being advised by public agencies
that the water was unsafe. By August 1978 practically all had ceased
potable use of the contaminated water. Thus, any abnormalities reflected by
the specimens collected in this survey would for the most part be due to
current exposure from washing and toilet uses and prior exposure from
drinking and cooking.
Population Characteristics--
A total of 36 exposed Hardeman County people were available for com-
parison with a total of 53 people in the Memphis control group. These groups
were composed of 14 males and 22 females from Hardeman County, and 23 females
and 30 males from Memphis. All participants were white. An age and sex
42
-------
profile for the two groups is shown in Table 12. Four additional Hardeman
County residents provided urine specimens only.
Liver Function Tests—
The results of the comparisons of the Hardeman County and Memphis group
are presented in-Tables 13 and 14. Significant differences were found for
several of the tests, alkaline phosphatase, S6PT and SGOT levels were
significantly higher in the Hardeman County specimens (p = .017, .039 and
.0001, respectively). Albumin and total bilirubin were lower in the Hardeman
County population (p = .0002 and .0001, respectively). Examination of the
variation of results for age and sex (Table 14) revealed that for SGOT the
geometric mean for Hardeman County males, 22.0, was at the upper limit of
the normal reference range of 8-22. The geometric mean for Hardeman County
persons under 12 years of age was 23.4 was slightly above the limit for
the normal range and that for persons 12-21 years was slightly below this
limit, 21.6.
Analysis of Urine Samples for Selected Organic Compounds--
In November 1978, urine specimens from 96 participants (43 exposed
Hardeman County residents and 53 Memphis controls) were screened for five
chlorinated organics HEX, HEX-BCH, heptachlorobicycloheptene, chlordene
and heptachlor. None of the samples from the Hardeman County residents
showed the presence of any of these substances. However, urine from six
of the adult males in Memphis contained HEX-BCH in amounts ranging from
0.3 to 1.0 ng/ml, probably due to the proximity of their location of work
to the pesticide manufacturer.
Environmental Data--
Environmental data from air and water samples collected from several
locations in the two communities are presented in Tables 15 and 16. Air
and water samples were not taken in the homes of the Memphis families used
in these comparisons. However, such sampling was performed elsewhere in
Memphis at about the time of the collection of their blood and urine specimens.
Of particular interest is the air data for August 23, which was taken in
the bathroom of one residence in Hardeman County, the first sample was
collected with no water running and the second with the shower on and the
door closed. Levels of carbon .tetrachloride increased by over a factor of
100 between these two samples. The higher value is approximately five percent
of the ACGIH recommended occupational standard for work place air (38). The
reduction in tetrachloroethylene (TCE) in the sample taken with the shower
running may be due to more effective collection and retention of carbon
tetrachloride by the sampling sorbent or displacement of TCE by carbon tetra-
chloride in the sampling material. The only air sample during this time
period which was associated with a noticeable air odor was the August 23,
1978 sample collected with the shower running.
Detectable levels of HEX in air were found in three of the five Hardeman
County home samples in November 1978. The highest concentration was 0.10
/Jg/m^. in comparison, air levels at the Memphis North Treatment Plant
ranged up to 39 and 278/jg/m3 for HEX and HEX-BCH, respectively. Concentration
of these same two compounds in urine collected from treatment plant workers
ranged up to 3.9 and lO.Sjug/L, respectively (39).
43
-------
TABLE 12. AGE AND SEX PROFILE OF HARDEMAN COUNTY EXPOSED STUDY
GROUP AND MEMPHIS CONTROL GROUP (NOVEMBER 1978]
Age, Range Years
Location and Sex
Less than 12 years
12-21 years
More than 21 years
Totals
Hardeman County
Males Females
5
2
_7
14
7
4
r\_
22
Memphis
Males
2
5
23
30
Females
6
7
I°_
23
44
-------
TABLE 13. HEPATIC PROFILE COMPARISON OF HARDEMAN COUNTY EXPOSED GROUP
AND MEMPHIS CONTROL GROUP (NOVEMBER 1978)
»
Parameter
Alkaline Phosphatase
(32-72 mU/ml adults 21
25-150 mU/ml
childreru21)
»•
Serum gamma glutamic
transaminase (SGGT)
(5-29 mU/ml)
Albumin
(3.5-5.0 gr/dl)
Total Bilirubin
(O.l-l.i mg/dl)
Serum glutamic pyruvic
transaminase (SGPT)
(5-25 mU/ml) '
Serum glutamic oxalic
transaminase (SGOT)
8-22 mU/ml) :
Hardeman County Memphis
Mean**-
, Range
No. Above
Normal
Total Tested
Mean*
Range
No. Above
Normal
Total Tested
Mean
Range.
No . Above
Normal
Total Tested
Mean*
Range
No. Above
Normal
Total Tested
Mean*
Range
No. Above
Normal
Total Tested
Mean*
Range
No. Above
Normal
Total Tested
88". 1
34-360
17/36,
9.47
2-54
3/36 '
4.35
3.9-4.8
0/36
,240
0.1-0.8
0/31
15.9
9-50
5/36
19.5 ,
12-36
11/36
59.9
29-191
6/53
11.23
4-68
4/51
4.53
3.7-5.7
16/49
.-:.-. .490 .. .
0.1-2.0
3/40
13.3
4-60
3/53
14.9
3-35
8/51
Significance
of
Difference
(t-test)
.017
.574
.0002
.0001
"•..'-
.039
.t)001
*Normal range indicated in parenthesis
* *Geometric mean
45
-------
TABLE 14. VARIATION IN HEPATIC PROFILE WITH AGE AND SEX, HARDEMAN COUNTY - MEMPHIS
COMPARISON (NOVEMBER 1978)
(Mean Values)
Age
Less Than 12 Years 12-21 Years
More Than 21 Years
Hardeman Co. Memphis Hardeman Co. Memphis Hardeman Co. Memphis
Age (N)
Alk. Phos.* (N)
SGGT* (N)
Albumin (N)
Total Bili.* (N)
SGPT* (N)
SGOT* (N)
(12)
(12)
(12)
(12)
(11)
(12)
(12)
9-1 (S)
187.3 (8)
7.28 (8)
4.35 (8)
0.21 (3)
15.4 (8)
23.4 (8)
8.5 (6) 15.8
151.7 (6) 94.6
5.90 (6) 6.17;
5.24 (6) 4.48
0-16 (5) 0.20
18.2 (6) 19.2
21.8 (6) 21.6.'
(12)
(10)
(10)
(10)
(7)
(12)
(12)
16.9 (18)
63.2 (18)
8.87 (18)
4.97 (18)
0.40 (15)
13.8 (18)
13.2 (18)
37.0 (33) 38.5
52.0 (33) 47.1
.13,0 (33) 14.1
4.30 (33) 4.23
0.28 (30) 0.57
15.3 (33) 12.2
16.9 (33) 14.2
Sex
Males
Hardeman Co. Memphis
Age (N)
Alk. Phos.* (N)
SGGT* (N)
Albumin (N)
Total Bili.* (N)
SGPT* (N)
SGOT* (N)
(14) 26.5
(14) 87.5
(14) 10.2
(14) 4.40
(12) 0.20
(14) 18.8
(14) 22.0
(30) 34.1
(29) 56.4
(29) 16.1
(29) 4.20
(26) 0.60
(30) 14.1
(30) 15.2
Females
Hardeman Co. Memphis
(22) 22.7
(22) 88.5
(22) 9.04
(22) 4.31
(19) 0.27
(22) 14.4
(22) 18.1
(23) 22.6
(22) 65.0
(22) 7.00
(22) 4.97
(14) 0.33
(23) 12.4
(?3) 14.4
*Geometric mean
(N) = Number of individuals in group.
-------
TABLE 15. CONCENTRATIONS OF SELECTED ORGANIC COMPOUNDS IN AIR SAMPLES (1978)
Sample Location
Hardeman County Exposed Group
Residence 1 - bathroom
(shower off)
Residence 1- bathroom
(shower on)
Residence 1 - kitchen
Residence 2 - bathroom **
Residence 3 - living room
Residence 4 - kitchen
Residence 5 - living room
Other;
Date
8/23/78
8/23/78
11/6/78
11/6/78
11/6/78
11/6/78
11/6/78
Memphis Residences 9/5-9/8/78
Memphis Industrial Area ***
Memphis Library
Memphis Residence
9/6/78
9/13/78
11/6/78
HEX
.07
n.d.
0.10
.09
n.d.
.06-
n.d.
n.d.
n.d.
n.d.
n.d.
HEX-BCH
.05
0.5
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
0.05
n.d.
n.d.
HEX-VCL*
n.d.
n.d.
n.d.
n.d.
n.d.
0.6
n.d.
n.d.
0.3
n.d.
n.d.
Chlordene
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
CClj
23
3600
3.6
10
8.8
36
41
n.a.
n.a.
n.a.
1.2
Tetrachl oroe thyl ene
302
120
3.6
2.4
2.5
5.7
n.d.
n.a.
n.a.
n.a.
2.7
n.d. » none detected
n.a. = not analyzed
*Heptachlorobicycloheptene
** Also contained heptach1or(2.2 ng/1/
*** located near pesticide manufacturer
-------
-p.
00
TABLE 16. CONCENTRATIONS OF SELECTED ORGANIC COMPOUNDS IN WATER SAMPLES (1978)
Sample Location
Hardeman County
Residence 1
Pugh Creek
Residence 1
Other:
vug/1
Date HEX HEX-BCH HEX-VCL Chlordene CC14
Exposed Group
10/2/78 0.72 0.25 n.d. n.d. n.a.
10/2/78 n.d. n.d. n.d. n.d. n.a.
11/7/78 n.d. 0.06 n.d. n.d. 618
Three residences (two in 10/2/78 n.d. n.d. n.d. n.d. n.a.
Memphis and one in Bolivar, TN)
n.d. = none detected
n.a. = not analyzed
-------
Medical Histories--
Medical questionnaires focused on illnesses during the 12 month period
prior to the study, revealed that there were 11 hospitalizations of the
Hardeman County residents. Symptoms included gastrointestinal abnormalities,
respiratory difficulties, neurological symptoms, muscular complaints, and
high fever. Many other persons reported becoming ill with symptoms such as
dizziness, headache, skin rashes, nausea and vomiting, numbness of limbs,
and menstrual irregularities. During this same time period the only birth
was a low birth weight infant with gastrochisis and eventration of intestines.
Seven adults and five children reported vision problems.
Summary
This survey revealed that concentrations of the enzymes alkaline phos-
phatase,SGPT and S60T were statistically higher in the residents who drank
the contaminated water group than in the control group. This difference
existed even though the adult males from the Memphis control group were
exposed to some air contamination by the "HEX compounds" as revealed by the
air sample from a,Memphis industry (Table 15) and by the presence of HEX-BCH
in some of the specimens. The differences appear to be associated with well
water contaminated by the toxic waste landfill. Residents and others have
noticed chemical-Tike odor emanating from the dump site during the period of
its operation and on more recent occasions indicating that the exposure
undoubtedly was not due entirely to the direct contact with contaminated
water. Based on the results of the November 1978 survey it was also con-
cluded that a more extensive survey was warranted.
Results - January 1979
Water Use—
At the time of this study all persons in the exposed group were using
the temporary piped water supply. Two families in the intermediate-exposure
group were using well water containing about 50 ppb of carbon tetrachloride
(36). Carbon tetrachloride was not detected in any other water supplies
tested (see Environmental Monitoring Results).
Population Characteristics—
The age and sex distribution for the exposed, intermediate-exposed and
control groups are presented in Table 17.
Liver Function Tests-
Results of comparisons between the January 1979 study groups appear in
Table 18. There were no significant differences detected between groups for
the liver function tests. The 30 people who were tested in both November
1978 and January 1979 showed a statistically significant change for all six
variables with alkaline phosphatase, S66T, S6PT and S60T all decreasing from
November 1978 to January 1979 while albumin and total bilirubin increased
(Table 19). The mathematical transformations used in the statistical analyses
of liver function test data and the significant covariables are presented in
49
-------
TABLE 17. AGE AND SEX PROFILE OF EXPOSED, INTERMEDIATE-EXPOSED
AND CONTROL GROUPS IN HARDEMAN COUNTY STUDY
(JANUARY 1979)
Age
0-14
15-24
25-34
35-44
45-54
55-64
65 and over
Total
Exposed
Male Female
6
2
5
3
0
3
1
20
13
3
8
1
1
1
2
29
Intermediate-Exposed
Male Female
3
0
5
0
0
1
1
10
12
2
7
0
0
1
1
23
Control
Male Female
4
1
7
3
2
1
1
19
7
4
10
4
2
7
3
38*
*0ne missing age
50
-------
TABLE 18. HEPATIC PROFILE COMPARISON
INTERMEDIATE-EXPOSED AND CONTROL
OF HARDEMAN COUNTY EXPOSED i
GROUP (JANUARY 19791
Groups
Exposed Intermediate-exposed Control
Alkaline
phosphatase
SGGT
Albumin
Total
bilirubin
Direct
bilirubin*
SGPT
SCOT
Mean
Range
No. above
normal
Total tested
Mean
Range
No. above
normal
No. tested
Mean
Range
No. above
normal
Total tested
Mean
Range
No. above
normal
Total tested
Mean
Range
No. above
normal
Total tested
Mean
Range
No. above
normal
Total tested
Mean
Range
No. above
normal
Total tested
76.33
29-450
18/48
12.27
5.51
6/48
4.80
3.0-7.6
16/48
0.49
0.2-1.4
4/47
0.12
0.1-0.2
0/47
12.45
2-49
1/48
16.91
10-41
11/48
79.02
34-186
15/33
10.32
5-35
1/33
4.81
4-5.6
9/32
0.51
0.2-1.6
3/28
0.12
0.1-0.2
0/26
14.08
7-27
1/33
16.31
11-25
5/33
61.49
31-220
8/56
11/56
4-56
3/56
4.93
4.2-6.2
23/57
0.51
0.2-1.7
4/52
0.13
0.1-0.2
0/52
14.25
6-70
4/56
16.08
9-140
7/56
*Normal range 0.1 - 0.4 mg/dl
51
-------
01
I\J
TABLE 19. COMPARISON OF HEPATIC PROFILE TEST RESULTS FOR EXPOSED -
PARTICIPANTS IN BOTH NOVEMBER 1978 AND JANUARY 1979 STUDIES
Variable
Hepatic Profile:
Alkaline phosp.
SGGT (log (5-SGGT)
Albumin
Total bilirubin
SGPT
SCOT
N
31
3]
31
26
31
31
Mean Difference*
13.3
-2.6
-.5
-.2
3.6
4.4
Standard
Error
^6.3
1.1
.13
.03
1.2
1.0
2
2
3.
6.
3.
3.
t
.125
.302
,922
405
044.
989
Degree of
Freedom
30
30
30
25
30
30
i
P
P'< .0419
P<.0284
p<.0004
P < .0001
p< .0048
P< .0003
*November results minus January results.
-------
Table 20 along with similar information for the bile acid and renal function
test data.
Bile Acids-
Results of analyses of fasting and nonfasting serum and urine samples
for specific bile acids - sulfated conjugates of lithocholate (SLCC) and
cholyglycine (CG) appear in Table 21. The mean nonfasting serum SLCC values
were significantly lower in the exposed group than in the control, however,
the clinical significance of this finding is unknown, since SLCC increases
in most liver diseases.
The mean change in SLCC and CG from the fasting to the nonfasting serum
specimens was positive for all groups as expected, however, in urine it was
negative for the exposed group and positive for the other two. These changes
in urine CG were statistically different for the exposed and control groups.
Bile acid determinations were also made on the nonfasting serum samples
collected from the exposed group in November 1978. Twenty-two of these in-
dividuals were retested in January 1979. A comparison of the November 1978
and January 1979 analyses (Table 22} revealed that serum concentrations
for SLCC were significantly higher in November 1978 than in January 1979.
Renal Function Tests--
Results of comparisons between the January 1979 exposed and control
groups appear in Table 23. Differences were found for total carbon dioxide
with the exposed group having somewhat lower total carbon dioxide levels
than the control group. Renal function tests were not performed on the
intermediate-exposed group.
Hepatitis-.Serology—
In order to determine if the results of the tests of liver function
were due to a prior history of infection with hepatitis, antibodies to hepa-
titis A and hepatitis B were determined. A log-linear model, taking age
into account, was used to determine that the incidence of positive hepatitis
serology was not significantly different between the groups.
Physical Examination—
During the physical examination 7 individuals were found to have border-
line enlargement of their livers, 6 being in the exposed group and 1 in the
intermediate group. Discriminant analysis was used to determine whether
there were differences in liver enlargement among the exposed, intermediate-ex-
posed and control groups. A significant difference between the groups was
found.
Illness--
Study participants were asked if they had any "serious illnesses" in
the prior six months. The subjective complaints were classified as either
respiratory, gastrointestinal or renal. Comparison of the results of the
illness interviewing (Table 24) indicates borderline differences for respiratory
illnesses (p =0.08) and all illnesses combined (p = 0.06). In each case
the intermediate-exposed groups had a higher illness rate than the exposed
groups which had slightly higher rates than the control groups.
53
-------
TABLE 20. TRANSFORMATION AND SIGNIFICANT COVARIABLES IN THE STATISTICAL
ANALYSES OF LIVER FUNCTION, BILE ACID AND RENAL FUNCTION TEST DATA
Variable
Transformation Used
Liver Function Tests
Alkaline phosphatase
SGGT
Albumin
Total bilirubin
Direct bilirubin
SGPT
SGOT
Serum Bile Acids
SLCC-fasting
SLCC-nonfasting
SLCC-difference
CG-fasting
CG-nonfasting
CG-difference
Urine Bile Acids
log
log
log
log
log
log
log log
log
log
square root square
root
square root
Significant Covariabjes^
age
age, sex
age, sex, age-sex
interaction
sex
sex
bLCC-fastinq
SLCC-nonfasting
SLCC-difference
CG-fasting
CG-nonfasting .:
CG-difference
Renal Function Tests
Na
K
Total C02
Glucose
BUN
Creatinine
square root
square root
log
square root
square
log
square root
— m.
log
age, sex, age -sex
interaction
sex
age, monthly alcohol
consumption, age-
alcohol consumption
interaction
age
age
age, sex
54
-------
01
en
TABLE 21. COMPARISON OF MEAN BILE ACID LEVELS (f STANDARD ERROR) IN
FASTING AND NONFASTING SERUM AND URINE SAMPLES COLLECTED FROM
PARTICIPANTS IN JANUARY 1979 STUDY
Serum Bile Acids
SLCC-fasting
SLCC-nonfasting
SLCC-difference
CG- fas ting
CG-nonfasting
CG-difference*
Urine Bile Acids:
SLCC-fasting
SLCC-nonfasting
SLCC-difference
CG-fasting
CG-nonfasting
CG-di f f erence*
Mean
0.769 +
1.106 +
+ .466 +
0.268 +
1.229 +
+ 1.987 +
1.607 +
1.513 +
- .078 +
.660 +
.645 +
- .028 T
Exposed
Standard
Error
.091 (48)
.078 (42)A
.071 (42)
.046 (48)
.135 (42)
.123 (42)
.209 (41)
.181 (44)
.155 (38)
.095 (44)
.074 (44)
.-101 (41 )B
Groups
Intermediate-Exposed
Control
Standard
Mean
0.878 +
1.490 T
+ 0.628 +
.255 +
1.0817
+ 0.861. +_
1.781 +
1.712 +
+ .054 +
C.635 T
0.766 +
+ .125 T
Error
0.136 (
0.220
,157
0.053
0.133
29)
24)
24)
?9)
24)
,157 (24)
.216 (28)
0.273 (24)
.204
.140
.126
.082
24)
(28)
(25)
(25)
Mean
0.843 +
1.526 +
+1.683 +
0.382 +
1.277 +
+ .906 ±
1.825 +
2.047 +
+0.281 +
0.521 +
0.^04 +
+0.288 +
Standard
Error
.065 (59)
.109 (58)A
.620 58)
.062 59)
.102 57)
.104 (57)
.187 (56)
.171 (51)
.135 (50)
.070 (55)
.072 (54)
.063 (52)B
Values in parenthesis are number of observations.
Letters depict significance between groups at p^-0.02.
*Nonfasting result minus fasting result.
SLCC - sulfated conjugates of lithocholate
CG - cholyglycine
-------
en
TABLE 22. COMPARISON OF SERUM BILE ACID LEVELS IN NOVEMBER 1978
SAMPLES WITH THOSE IN SERUM SAMPLES COLLECTED
FROM THE SAME INDIVIDUALS IN JANUARY 1979
Serum bile Acids:
SLCC-nonfasting
(log (SLCC +
1.5))**
CG-nonfasting
N
23
23
Mean
Difference*
0.22
0.32
Standard
Error
0.09
0.23
t
2.475
1.401
Degrees of
Freedom
22
22
P
p<.0211
p >. 1745
*November results minus January results.
**For SLCC the data required a log transformation of the serum concentration
plus a constant in order to produce a normal distribution.
-------
TABLE 23. COMPARISON OF MEAN VALUES (± STANDARD ERROR) OF RENAL
PROFILE TEST RESULTS BETWEEN EXPOSED AND CONTROL GROUPS
PARTICIPATING IN JANUARY 1979 STUDY
Renal Profile*
Sodium (Na)
(133-145 mEq/1)
Potassium (K)
3.5-5.0 mEq/T)
Chlorides (Cl)
(95-110 mEq/1)'."
Total Carbon Dioxide
(Total CO?)
(21-27 mEq/1)
Glucose
(60-95 mEq/1 )
Blood Urea Nitrogen
Groups
Exposed
Standard
Mean Error
141.97 + .51 (35)
4.606 '+ .077 (35)
103.50 + .38 (32)
23.13 .+ .96 (32)A
. 77.68 ±2.35 (34)
13.38 V .75 (34)
Control
Standard
Mean Error
141.26 + .53 (42)
4,54 + .07 (42)
104.59 + .54 (41)
26.27 + .64 (41 )A
82.05 +1,77 (42)
16.48 + .93 (42)
(BUN)
( 5-17 mg/dl)
Creatinine
(0.7-1.4 mg/dl)
0.72 + .03 (34)
0.84 + .03 (42)
Letter depict significant differences between groups (^-0.02)
*Normal ranges indicated in parenthesis.
57
-------
TABLE 24. COMPARISON OF REPORTED SERIOUS ILLNESSES IN PRIOR SIX MONTHS
(PERCENT SHOWN IN PARENTHESIS)
Respiratory
Number with illness
Number without illness
.Exposed
6(11)
47(89)
X? = 5.23, p = O.OS
•
Gastrointestinal
Number with illness 3(6)
Number without illness 50(94)
X? = 0.88, p = 0.65
groups
Intermediate-Exposed
10(23)
33(77)
4(9)
39(91)
Control
5(8)
56(92
3(5)
58(95)
Renal
Number"with illness 1(2)
Number without illness 52(98)
X2 = 0.23, p = 0.88
H2)
42(98)
2(3)
59(97)
All Illnesses Combined
Number with illness 10(19)
Number without illness 43(81)
X2 = 5.74, p = 0.06
13(30)
30(70)
7(12)
54(88)
58
-------
Extensive review of the medical histories obtained at the time of the
indepth investigation as well as review of hospital records supplied by
patients who had undergone hospitalization in the last several years was
performed. Hospital records showed no indication of any disease trend patterns
or clustering of diseases that could be attributed to consumption of con-
taminated water. Many residents, however, did report nausea, vomiting and
mucous membrane irritation and associated these symptoms with an unusual
chemical-type odor in the water used for consumption.
Analysis of Urine Samples for Selected Organic Compounds--
Urine specimens from 137 participants from the three groups of participants
were screened for the presence, of HEX, HEX-BCH, hexachlorobicycloheptene,
chlordene and heptachlor. None of the samples showed the presence of the
selected chlorinated organics. A select group of 9 individuals judged to
have relatively high potential for exposure to carbon tetrachloride were
sampled for determination of carbon tetrachloride in urine. Of the 9 persons,
5 were from the exposed group and 4 from the intermediate-exposed group.
These intermediate-exposed participants included individuals who were using
water containing about 50jug/l of carbon tetrachloride. No evidence was
found for carbon tetrachloride in any of these urine specimens. These findings
with human urine specimens is consistent with animal studies (40,41) which
show that carbon tetrachloride administered to mammals is largely excreted
through the lungs,.that only a small fraction of absorbed material is meta-
bolized, and that it is not excreted as the parent compound in the urine.
The primary metabolites include chloroform, hexachloroethane, and carbon
dioxide. It was, therefore, concluded that analysis of additional urine
specimens;, for carbon tetrachloride was not warranted at this time.
Environmental Monitoring--
Representative water samples were analyzed from all of the sources of
domestic water for the homes of the study participants. The monitoring was
conducted in both January and March 1979. The March monitoring included
some of the same homes as those sampled in January. For a number of the
homes indoor and/or outdoor air samples were collected.
Tables 25 and 26 provide a comparison of air and water samples col-
lected in January 1979 and March 1979, respectively for the exposed, inter-
mediate-exposed and control groups. Water samples were designated as tem-
porary (T), individual wells (I) and public (P). A temporary water supply
was in use for many homes along the Toone-Teague Road including all homes of
exposed participants and some homes of the intermediate-exposed participants.
Inside air samples were collected in the bathrooms with the shower or bathtub
tap running. The water samples containing the highest concentrations of
carbon tetrachloride and tetrachloroethylene were from the group one and two
homes. In general, the samples from the control group had typical back-
ground levels (42,43). Of the six control homes using a public water supply
one had an inside air sample containing greater than 1 >ug/m3 of carbon tetra-
chloride. Low levels of carbon tetrachloride, up to l.l^ug/L, were detected
in some of the public water supply samples which were routinely chlorinated.
59
-------
TABLE 25. CONCENTRATION OF SELECTED ORGANIC COMPOUNDS IN WATER SAMPLES AND
Group
and
sample
Exposed
R-ld
R-6
Type of
Water
Supply'3
T
T
Water Concentration
yug/l
Carbon
ND
ND
L '
Tetrachloride
Air Concentration /tig/ro3
Outside0 Inside^ .
74.6 0.6
0.5 6.3
Intermediate-Exposed
R-7
R-8.
R-9f
R-10
R-ll
R-12
R-13
Control
I
I
I
I
I
I
I
R-14
R-15
R-16
R-17
R-18'...
R-19
R-20
R-21
R-22
R-23
R-24 to
R-339
I
I
I
I
I
I
P
P
P
P
I
Exposed
R-l T
R-6 T
Intermediate-Exposed
R-7
R-8
R-9
R-10
R-ll
R-12
R-13
I
I
I
I
I
I
I
ND
ND'
ND
3.7
0.7
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
Tetrachloroethylene
ND
ND
ND
ND
ND
ND
ND
ND
ND
(continued)
0.4
NSe
NS
NS
NS
NS
NS
NS
0.
NS
0.
NS
1.
0.
0.
0.
NS
NS
0.4
ND
0.3
NS
NS
NS
NS
NS
NS
176
NS
NS
NS
NS
NS
NS
NS
ND
NS
0.2
0.3
NS
0.4
0.2
ND
NS
NS
1.2
1.0
11.6
NS
NS
NS
NS
NS
NS
60
-------
TABLE 25 (continued)
Group Type of
and Water Water Concentration Air Concentrationyug/m3
sample Supply" jug/L Outsidec Inside0
Control
R-14
R-15
R-16
R-17
R-18
. R-19
1 R-20
R-21
R-22
R-23
R-24 to
R-33
I
I
I
I
I
I
P
P
P
P
I
ND
ND
ND
ND
ND
ND
ND
ND
ND
NO
ND
NS
0.3
NS
0.3
NS
0.5
0.2
0.2
0.2
NS
NS
NS
0.3
\J m *J
NS
0.4
0.6
NS
0.2
0.2
0.2
NS
NS
aAll samples were analyzed for: hexachlorocyclopentadiene (HEX), hex-
achlorobicycloheptadiene (HEX-BCH), heptachloronorbornene, chlordene,
heptachlor, carbon tetrachloride and tetrachlorethylene.
The types of water supplies in use were: T - temporary supply provided for
homes with contaminated wells; I - individual wells; and P - public supply.
clnside samples were collected in a bathroom with water running. Outside
samples were collected within 10 meters of the house.
dThe outside air sample also showed 3.2/jg/m3 of HEX-BCH.
eNS - indicates that no sample was obtained.
*
This household was using a hand carried water supply, source unknown,
for potable purposes.
9Water samples only were obtained from these ten residences in the control
group. None of the substances analyzed in this study were found in any
of these samples.
61
-------
TABLE 26. CONCENTRATION OF SELECTED ORGANIC CHEMICALS IN WATER AND AIR
SAMPLES FOR RESIDENTS OF STUDY PARTICIPANTS (MARCH 1979)a
Group
and
sample
Exposed
R-l
R-6
R-346
Type of
Water
Supply^
T
T
I
flater Concentration
>ug/L
Carbon Tetrachloride
0.3
ND
29,450
i/.
Air Concentration/jg/fn01'
Outside
2.1
3.6
326
Inside,^.
1.4
31.8
NSf
Intermediate-Exposed
R-7
R-9
R-10
R-n
R-35
R-36
'• R-37
R-38
Control
I
I
I
I
I
I
I
P
ND
ND
96
96
0.3
ND
ND
ND
9.3
NS
3.1
19
0.7
ND
ND
NS
1.6
31.09
139
681
NS
1.8
0.6
0.7
R-19 .
R-20d
R-23
R-39
R-40J
R-41d
Exposed
R-l
R-6
R-34
I
P
P
P
I
P
T
T
I
Intermediate-Exposed
R-7
R-9
R-10
R-ll
R-35
R-36
R-37
R-38
I
I
I
I
I
I
I
P
ND 11.2
1.1 13.5
0.8 1.2
ND 1.3
ND ND
0.7 1.4
Tetrachloroethylene
0.8 1.4
ND 1.2
9.0 1.2
ND 2.0
ND NS
6.4 3.7
2.5 7.2
0.2 2.9
ND ND
ND 1.9
NO NS
(continued)
62
ND
63.1
ND
0.8
0.6
ND
3.3
8.0
NS
1.7
14.39
1.0
1.1
NS
2.7
2.6
3.0
-------
Group
and
sample
Control
R-19
R-206
R-23
R-39
R-40
R-416
Type of
Water
Supplyb
I
P
P
P
I
- P ••,-..
Water Concentration Air Concentration>ug/m31
Aig/L Outside Inside
ND
1.2
0.4
ND
ND
1.4
3.0
11.8
1.4
1.6
2.1
1.1
0.7
17.0
40.8
2.0
0.8
24.5
aAll samples were" analyzed for: hexachlorocyclopentadiene (HEX), hexachloro-
bicycloheptadiene (HEX-BCH), heptachloronorbornene, chlordene, heptachlor,
carbon tetrachloride and tetrachloroethylene.
bT - Temporary supply; I - Individual wells; and P - Public supply.
clnside samples were collected in a bathroom with water running. Outside
samples were obtained within 10 meters of the house.
'•'This sample was obtained from a well recognized to be contaminated and
thus no longer in use The sample was also found to contain 2.5jug/L
HEX-BCH and 390 mg/L chloroform. The air sample, collected during and
after water was drawn from the well, showed 0.6jug/m3 HEX-BCH.
indoor air samples had 4.6 and 1.5jug/m3 of heptachlor for R-20 and
R-41, respectively. The source is unknown.
- Indicates that no sample was obtained.
9An air sample from a second bathroom in this home contained only 0.8>og/m3
of carbon tetrachloride and 0.6/jg/m3 of tetrachloroethylene.
63
-------
No carbon tetrachloride was detected in any Individual wells that were not
on the Toone-Teague Road.
A summary of the air and water sampling results for January and March
1979 combined are presented in Table 27. Carbon tetrach'Toride and tetra-
chloroethylene were detected in most air samples, regardless of the group
tested. Individual private well samples, except those along Toone-Teague
Road did not contain detectable amounts of either compound.
DISCUSSION AND, CONCLUSIONS .
The concentrations of the suspected carcinogen carbon tetrachloride
found in the contaminated well water (up to about 30,000 ppb) are far above
the level of 2.6 ppb that has been proposed as a safe level for water (42).
A survey of 80 municipal .water supplies conducted by the U.S-. Environmental
Protection Agency in 1975 revealed that about 10% of the supplies had de-
tectable levels of CC14 ranging from 2 to 3 /jg/L (ppb) (40). Another study
of 113 public water supplies reported in 1977 confirmed the earlier survey
(10% positives range; 2. 4-6.4 /jg/L) (41). Concentrations of carbon tetra-
chloride in the public water supplies used by some of the control group in
this study ranged up to 1.0 ppb. The releases of CC14 into the Ohio River in
Feburary 1977 resulted in surface water concentrations as high as 340/jg/L
(ppb) (42,43). Concentrations in the contaminated wells have frequently
been of the order of 5,000/ig/L. The normal background atmospheric concen-
trations of CC14 found in the continental air mass ranges from 0.78 to 9.1
^ig/m3 (ng/1) (42,44). Although some urban areas experience higher levels;
for example, in Bayonne, N.J. levels as high as 117Aig/m3 have been found
(45) /.The highest level observed in. an outside sample in this study was 326
Carbon tetrachloride :is regarded as a highly toxic :substance (46). It
is absorbed through the lungs, gastrointestinal tract, and. the skin. Although
gastrointestinal and hepatorenal injuries and nervous system symptoms result
from both ingestion and inhalation, gastrointestinal and hepatorenal in-
juries are most prominent in poisoning from ingestion, and nervous symptoms
predominate in inhalation poisoning. In all poisonings the liver is the
most drastically affected organ but kidney injury may be more prominent
clinically. The lowest concentration in air that has produced any toxic
effect in man is 130 mg/m3.
Lauria and Bogden (47) have recently reviewed the adverse effects from
limited exposures to CCla. They concluded that exposure to low levels of
CC14 in air (63-157 mg/m3) is not toxic but that at greater exposure concen-
trations, nausea, anorexia, headache, dizziness, lassitude, and vomiting
were observed. Liver function abnormalities (elevation of transaminase and
LDH) were also noted. The symptoms disappeared promptly when removed from
the source of exposure. At increased exposures, nausea and vomiting were
more prominent, accompanied by severe abdominal pain. It is also believed
that any exposure to air concentrations above 628 mg/m3 can be asssociated
with severe liver and/or kidney damage, and that far less severe disease may
be induced by exposures over 157 mg/m3. The review also showed that acute
liver damage can be induced readily in a variety of animal species. The
64
-------
TABLE 27. COMPARISON OF WATER AND AIR SAMPLES OF RESIDENCES
OF STUDY PARTICIPANTS (JANUARY AND MARCH 1979)
Group
and
Exposed
7.
I
Carbon
Water
(Range) c
1/4
(0.3)
1/1
(29,450)
Tetrachloride3
Air Samples
Outside
(Range)
4/4 d
(0.5-75)
1/1
(326) '
Inside
Np/r
(Range)
4/4
(0.6-31.8)
NS
Tetrachloroethylene2
Water
(Range)
1/4
(0.8)
.1/1-
(9.0)
Air Samples
Outside Inside
(Range) (Range)
3/4 4/4
(1.2-1.4) (3.3-8.0)
1/1 NS
(1.2) NS
Intermediate-Exposed .
I '
P
Control
I
P
5/14
(0.3-96)
0/1
0/18
3/8
(0.7-1.1)
5/7
(0.4-19)
NS'
NS
4/5
(0.2-11.2)
7/7
(0.1-13.5)
8/8
(0.6-681)
1/1
(0.7)
3/5
(0.2-0.6)
4/7
(0.2-63.1
3/14
(0.2-6.4)
0/1
0/18
3/8
) (0.4-1.
5/7 8/8
(0.3-7.2) (0.6-14.3)
NS 1/1
NS ' (3.0)
5/5 4/5
(0.3-3.0) (0.3-0.8)
7/7 7/7
4) (0.2-11.8) (0.2-4Q.8)
aWater concentrations are ng/L; air concentrations are^jg/m3.
bT designates temporary water supply.; I - individual well; P - public supply.
CN, designates total number of samples collected; Np designates the
, number with detectable levels.
dOne air sample also showed 3.2^g/m3 of HEX-BCH.
eNS indicates no sample.
65
-------
s
«np S M! t0 be °rdlnarily repaired if the insult is limited in
dosage and the repair may be complete even if necrosis and fibrosis have
occurred From animal feeding studies, 10 mg/kg body weight per dav was
consld r d^ "f6 ?' the ^ and a d°Sage o? ^ PP-^
SS r. ;%!MedgSSds Yof^ £3? •iB$!d
from human experience did not indicate that exposureto sma 11 qwntitiw of
tt^toTS uce6 e:PlhVhreSh?ld ^°dUCing >*»*** necrosls'h ^ apa-
d d not indicate th*t rri " ?? 61ther liver or kidne^ Human experience
Academy of Scienc^/Lf?n4 !V ^^ carcin°9en. A committee of the National
ra/t^?^
Sl^V9e^ J" P^^MeSeTS i? ^g ^i ''ambient
eeexarft ° have been dumPed at the
(HEX-BCH ?he ccJS5I?nS 1° ^P6"*8^6"6 HEX) a™* hexachlorobicycloheptadiene
-
ara« Ks
and uper re p ^tory traci anS h^H h"6 exPerienced irritation of the eye
burns upon contact with thp'^n^H f I?es1upon exP°sure to HEX vapors; and
term exSosureS may result f n a^ lp/nf al Stud^es have Indicated that long
(51). There is no Smaiion avaitahf nonsP^lfic adverse health effects
chronic exposure to HEX The?e hill h re9ardln9 the health effects of
of «rkersPwho are occupatl ly' p *3 g ^°^ of ep1de»1olog1c studies
mat-ion available concerninq the ootPntHi K i!J (H"53)> There 1S no infor"
chronic environmental exposure tS HEX v'a ^na]^h.effe^S. resulting from
Even less is known of the toxic effects of S?y S^atKd.a1r and/or water' - c
that HEX-BCH may be as toxic as HFX (ll\ ^X-BCH> but there are indications
the Threshold Limit Value TLV) for HF^RPH" S?i 1* may be est1mated that
for HEX (0.1 mg/m3) Tn orirtivl h C? W0uld be about ec1ual to the TLV
two or more haSirtois b " SSlr^^V^H^0811™ l6V6lS ^
additive unless other information ic 1 ••> !^ts should be considered to be
for complex exposure station i e llll^ .™S practice 1mP1ies that
contamination of a groundSatS? suiil^bl ! S u** might be enc°unt^ed with
leachate, determination of S5o!u?e leSeU ^Ul-eomP°nent toxic dumP
UT exposure levels of a single chemical may not
66
-------
adequately reflect the actual exposure burden experienced by an individual.
In this case, consideration of the exposure levels of several substances is
recommended.
Exposure to many agents including drugs, alcohol, and a variety of
chemicals as well as disease states including hepatitis can cause liver
damage. However, based on the results of this study, which excluded many of
these potential causes, the conclusion is reached that the most likely reason
for the presence of liver abnormalities discovered in the inhabitants of the
Toone-Teague Road area in November 1978 was the consumption of water from
wells contaminated by the leaching of material from the chemical land dump
located in that area. The acute symptomatology and the abnormal liver function
tests have in all but a few cases returned to normal and with no further
contaminations, should remain normal. However, it is impossible at this
time to predict whether there will be any future chronic effects as a result
of this exposure. Six of the seven individuals found to have enlarged livers
were in the exposed group. A medical follow-up.of the people studied in this
investigation is warranted.
Preliminary toxicity and mutagenicity testing (55) has shown that con-
tamination of some of the well water samples was sufficient to cause a toxic
response and, in one, a slight mutagenic reponse. The well water samples
were tested for this biological activity without any concentration step in
the laboratory procedure. Municipal drinking water from Cincinnati or New
Orleans requires concentration of the organic components to elicit a bio-
logical response. HEX-BCH, reported to cause a mutagenic reponse in the
Ames assay (56), was detected in the contaminated well water as were the
suspected carcinogens, chloroform and carbon tetrachloride.
The water sample collected from a heavily contaminated well in March
1979 contained the highest concentration of carbon tetrachloride found in
that area up to that time. This result indicates that contamination of the
aquifer is increasing even with decrease in the overall usage of the aquifer
as a water supply. Future contamination of the much deeper artesian aquifer,
which is used at a location about 50 miles away by Memphis, Tennessee, is a
distinct possibility because abandoned well shafts to this aquifer could be
plugged at the present time and, therefore, not be detectable by the static
head tests performed by the U.S. Geological Survey (57).
67
-------
SECTION 6
EVALUATION OF WORKER EXPOSURE TO PESTICIDE CHEMICALS AT A
MEMPHIS, TENNESSEE, WASTEWATER TREATMENT PLANT
During a prospective study aimed at evaluating the health risks from
bacteria and viruses associated with the treatment of municipal wastewater,
an opportunity developed to investigate human exposures to toxic organic
chemicals emitted from the wastewater being treated at one of the plants in
this study. The particular plant in question was the North Wastewater Treat-
ment Plant in Memphis, Tennessee.
This plant, which began operation during the summer of 1977, is located
five miles from a manufacturer that produces and utilizes several chlorinatea
organic intermediates for the synthesis of flame retardants and pesticides
(notably isodrin, endrin, chlordane and heptachlor). Waste from this manu-
facturer is discharged into a sewer that flows to the Memphis North Treatment
Plant. Prior to construction of this treatment plant, wastewater from the
areas served by this plant flowed directly to the Mississippi River or to one
of its tributaries (60). Concern for potential exposure of treatment plant
employees to toxic chlorinated organics prompted an investigation (61) during
August and September 1977 to evaluate the potential hazards that might result
from the initial diversion of industrial wastewater to the newly completed
municipal wastewater treatment plant. The preliminary investigation revealed
that air in the wet well of the treatment plant caused eye irritation and had
a strong chemical odor similar to that at the pesticide manufacturing plant.
Concentrations of hexachlorocyclopentadiene (HEX) in the air samples analyzed
were below the detection limits of 0.008/jg/l for the analytical procedures
used. At the request of Region IV, U.S. Environmental Protection Agency, tne
National Institute of Occupational Safety and Health (NIOSH) made an evaluate'
of the potential problems associated with allowing HEX to flow into the Memphis
North Treatment Plant. Concern over exposing workers to this toxicant was
expressed, and it was recommended that, "The workers should not deliberately
be used as test subjects in evaluating the toxic hazard from HCCPD." (62)
HCCPD is an alternative abbreviation to hexachlorocyclopentadiene.
In the early part of 1978, following the initial evaluation, workers at
this same plant complained of acute symptoms of respiratory distress,
dizziness, headache, and irritation of the eyes, throat, nose, lung and skin.
It was again noted that the plant had an unusual odor characteristic of in-
dustrial chemical wastes. The incidence of these acute symptoms was reported
to be associated with periods of more intense chemical odor. In March 1978,
during an apparent slug release of the pesticide materials, six workers on
the night shift became ill with symptoms of loss of equilibrium, severe
68
-------
headaches, and nausea. Two were taken to a local emergency room, examined
and released with no definitive diagnosis.
Although it has been generally recognized that workers in sewage treat-
ment plants may be at an increased risk of exposure to infectious agents (1),
only recently have studies revealed incidences of exposure of sewage
treatment plant workers to potentially toxic chemicals (64-66). Workers
may have direct contact with chemical waste materials during solids removal
and other operations; however, the primary route of exposure of treatment
plant workers to these substances is probably by inhalation. Chemical wastes
enter the treatment plant either in a soluble form or adsorbed to suspended
materials in the wastewater. Aeration during the treatment processes can
cause aerial dispersion of these substances as aerosols and vapors.
The symptoms reported by the workers in the present study were similar
to those experienced by workers approximately a year earlier at the Morris
Forman Wastewater Treatment Plant in Louisville, Kentucky (64,65). In that
incident, exposure was determined to come from pesticide intermediates,
hexachlorocyclopentadiene (HEX) and octachlorocyclopentene (OCTA), which were
illegally dumped into the municipal sewer system. Ambient air levels of HEX
several days after the plant was closed ranged from about 3,000>jg/m3 to
10,000>ug/m3, which far exceeded the recommended maximum eight-hour time-
weighted average exposure level of 100jug/m3 (67). Because of the similar-
ities in these events in Louisville and Memphis, an investigation to determine
the degree of exposure to these chemical contaminants at the Memphis sewage
treatment plant was undertaken.
STUDY DESIGN
In view of the intermittent occurrence of acute symptoms experienced by
the workers, a study was undertaken during May to September 1978, to de-
termine if evidence of exposure to the toxic chemicals and absorption of same
could be detected by use of biological testing. For comparison purposes, a
second plant in the Memphis area, the Maxson Wastewater Treatment Plant, was
used. The sewer from the pesticide manufacturer was not connected to the
Maxson Plant so it was felt that exposure there should be low to nonexistent
from that route of entry. The location of these two wastewater treatment
plants and the pesticide manufacturer are shown in Figure 3. Three studies
were performed during the May-September 1978 period. The protocol for these
involved four elements which are illustrated in Table 28 and are elaborated
as follows: (1) illness symptom questionnaire, (2) analysis of urine
specimens for selected organic compounds, (3) liver and bile acid determinations,
and (4) air and wastewater monitoring. In addition, during July an oppor-
tunity arose to collect urine specimens from several groups of workers in-
cluding some workers at the North Plant during a period of chemical odors.
The initial objective of this study was to establish a baseline chemical
exposure, if any, of the wastewater treatment plant workers during normal
operational conditions so that if a spill incident occurred or employees
experienced acute symptoms an assessment of a potential chemical exposure
could be more readily made. Urine samples were collected during the second
69
-------
Figure 3. Location of Memphis Wastewater Treatment Plants.
pesticide manufacturer and Memphis Light, Gas
and Water.
70
-------
TABLE 28. BASIC ELEMENTS OF EXPERIMENTAL DESIGN
Time of Study
Element May 1978 June 1978 September 1978
Illness symptom
questionnaire X X X
Urine analysis -
early in shift X
late in shift X X X
Liver and bile acid
determinations X
Wastewater monitoring X X X
Air monitoring
area
personal
Groups Tested
Memphis North Plant
Maxson Plant
MLG&W Workers
Mill Creek Plant
X
X
X
X
X
X
X
X
X
. • ,. x.-, ....'
X
71
-------
half of the work shift in May 1978 for this purpose. In addition, a limited
environmental monitoring survey was performed to determine a) if the influent
wastewater contained selected organic contaminants, and b) if so, were these
substances being released into the air during the treatment processes.
Follow-up urine screenings were conducted during June and September
1978. Urine samples at both the start and end of the work shift were collected
in June 1978. During the September 1978 screening end of shift urine samples
were also obtained from a group of workers at the Mill Creek Treatment Plant
in Cincinnati, Ohio, and from workers at the Memphis Light, Gas and Water
(MLG&W) Company. The MLG&W workers served as a control population for in-
fectious disease studies of Memphis sewage treatment plant workers and thus,
were included in this study as a population of workers with no association
with the sewage treatment plants. The Cincinnati workers served as an
additional control. In conjunction with the collection of urine specimens
each employee was administered a questionnaire eliciting a number of symptoms
and potential work-related problems. The interview related to the May urine
collection was administered at the same time as that for June, requiring an
approximately 30-day recall. Liver function tests and serum bile acid
analyses were performed on blood specimens of participants in the ongoing
seroepidemiological study to aid in a determination of possible chemical
toxicity involving the liver.
During both June and September 1978 area air sampling was performed to
provide an estimate of exposure levels of workers and measurements were
made of the influent wastewater to determine the concentrations of selected
organic contaminants present. Personal air monitoring of a group of workers
at the Memphis North and Maxson wastewater treatment plants was conducted in
September 4978. Eight-hour samples were collected at their work site and
also while they were away from work.
Analytical Methods
The urine specimens were frozen immediately after collection and were
kept frozen until analyzed. The frozen urine specimens were thawed in a cold
water bath. A 5.0 mL aliquot of urine was placed in a screw cap tube, satu-
rated with sodium chloride and mixed vigorously for 5 minutes with 5.0 mL of
pesticide grade petroleum ether. The petroleum ether extract was dried over
anhydrous sodium sulfate and stored in a freezer until analyzed. The extracts
SSnJ!nlJZ?n nr HEX and hf achlorobicycloheptadiene (HEX-BCH) by gas chro-
fnlSS Sh( i } UST dn electron capture detector. The analyses were per-
formed with glass columns packed with OV-101 and OV-17/QF-1 at 175°C. Spehar
et al employed a similar extraction and GLC analysis procedure for measuring
samplesC(S7j °n °f HEX' HEX'BCH and ^Ptachlorobicycloheptene in water
consisted of drawing air through glass sampling tubes con-
°f .Pre-extrac1*d Chromosorb 102 (69,70). DesoVption of the
SiSS
72
-------
the same as the sampling and analysis procedure for HEX recommended by NIOSH
(71) except that Chromosorb 102 was used as the sorbent rather than Porapak
N. Commercially available Chromosorb 102 gave numerous peaks in the gas
chromatograms under the desorption and analysis conditions. Removal of these
impurities was achieved by successive extraction of the sorbent in a Soxhlet
extractor with methanol, acetone, and petroleum ether. The sorbent was dried
in a vacuum oven and pre-tested immediately before use.
The concentrations of the chlorinated organics in wastewater samples
were determined by saturating an aliquot of the wastewater with sodium
chloride, extracting with an equal volume of petroleum ether. A 5/iL aliquot
of the extract was injected into the gas chromatograph (72).
Statistical Methods
The distributions of the measurements of hexachlorocyclopentadiene and
hexachlorobicycloheptadiene were skewed to the right. Following the use of
the logarithmic (base 10) transformation, these variables now possessed
unimodal, symmetric, bell-shaped distributions. The numbers of symptoms were
transformed by taking their square roots. In order to evaluate the relation-
ships among the above two variables, the symptoms of interest, and the co-
variables age, race, sex, date of measurement, work shift, time of measurement
within work shift, distance from North Plant of residence, several analyses
of covariance were performed. The dependent variables were the symptoms of
interest and the above two variables. Each of these analyses of covariance
were done in the following sequential manner due to the fact that several
workers had one or more measurements of coyariables missing from the data.
Initially, all independent variables were included in the covariance model.
Following this first analysis, those independent variables which were clearly
nonsignificant (p?0.50) were excluded from the model. Following the second
analyses, those variables which were nonsignificant (p>0.25), were excluded
from the model. In the final analysis, those variables with p<0.05 were
taken to be significant.
RESULTS
Urine Analysis and Illness Symptoms
Age, race and sex characteristics of the participants in the May and June
1978 screenings are presented in Tables 29 and 30 and those for the September
1978 screening in Tables 31 and 32.
An examination of the results of the urine analyses for the May col-
lections, during the second half of the work shift only, revealed that con-
centrations of HEX-BCH in specimens from North Plant workers tended to be higher
than those from Maxson Plant workers (Table 33). No differences were detected
In the HEX concentrations (Table 34). HEX-BCH was present in 84% of the North
Plant samples and in only 34% of those from the Maxson Plant. Sixty- nine
percent of North Plant samples contained greater than 3;ug/l of HEX-BCH, while
the percent of South Plant samples in this concentration range was 10% (p ^
0.001). Concentrations of HEX in urine were not statistically different between
the two plants.
73
-------
TABLE 29. AGE DISTRIBUTION OF PARTICIPANTS IN MAY AND JUNE 1978
_^ URINE SCREENINGS
Age Range, Years
15-24 25-34 33-44 45-54
May 1978
North Plant*
Maxson Plant
Total
June 1978
North Plant**
Maxson Plant
Total
Number of individuals
tested at least once
North Plant
Maxson Plant
Total
9
4
13
20
3
23
23
4
27
28
19
47
27
12
39
29
23
52
2
3
5
6
4
10
7
6
13
1
3
4
4
2
6
4
3
7
1
0
1
1
3
4
1
3
__— i— —
. •— '
*Age not available for one person.
**Age not available for two persons.
74
-------
TABLE 30.
RACE AND SEX DISTRIBUTION.OF PARTICIPANTS IN MAY AND
en
North Plant
White
Black
Total
Maxson Plant
White
Black
Total
Both Plants
White
Black
Total
•— ^— •—
uun
— ii ..I
It. 1J/V '
Mav 1978
Male
30
6
36
16
7
23
46
13
59
Female
3
3
6
1
5
6
4
8
12
Total
32
9
42
:i7
12
29
50
21
71
..I i •
Male
43
10
53
10
9
19
53
19
72
—
June 1978
Female
2
5
7
1
4
5
3
9
12
Total
45
15
60
1 1
13
24
56
28
84
Number
tested
Male
48
10
58
19
14
33
67
24
91
of Individuals
one or both dates
Female
3
5
8
1
5
6
4
10
14
Total
51
15
60
20
19
39
71
&
-------
TABLE 31. AGE DISTRIBUTION OF PARTICIPANTS IN SEPTEMBER 1978
URINE SCREENING
North Plant
Maxson Plant
MLG&W
15-24
14
7
2
Mill Creek Plant** 5
All Locations
28
*Age of one participant
Age Range,
25-34 35-44
26 5
16 2
11 3
12 2
65 12
unknown.
Years
45-54 55-64
3 5
2 2
3 6
1 0
9 13
Total .
53
30*
25
20
128
**Screening occurred in October 1978.
TABLE 32.
RACE AND SEX DISTRIBUTION
OF PARTICIPANTS
IN
SEPTEMBER 1978 URINE SCREENING
Male Female
North Plant
Maxson Plant
MLG&W
Mill Creek
Plant*
All Locations
White
Black
Total
White
Black
Total
White
Black
Total
White
Black
Total
White
Black
Total
41
7
48
17
10
27
15
9
24
15
4
20**
88
30
119**
2
3
5
1
2
3
0
1
1
0
o
0
3
6
9
Total
43
10
53
18
12
30
15
10
25
15
4
20
91
36
128
*Screening occurred in October 1978,
**Race of one participant not known.
76
-------
TABLE 33. HEXACHLOROBICYCLOHEPTADIENE (HEX-BCH) CONCENTRATIONS IN URINE
SAMPLES OF MEMPHIS WASTEWATER TREATMENT PLANT EMPLOYEES
SECOND HALF OF WORK SHIFT. MAY 1978
Plant
North*
Southa
Total
Samples
32
29
Range
/ig/L Below
* 0.3-8. 4
<.0.3-7.5
Urine
Detection
5
19
Concentrations.
less than or
equal to 3
5
7
, -Ajg/L
greater
than 3
22
3
a North Plant results higher than South Plant results (p<,.001).
TABLE 34. HEXACHLOROCYCLOPENTADIENE (HEX) CONCENTRATIONS IN URINE
SAMPLES OF MEMPHIS WASTEWATER TREATMENT PLANT EMPLOYEES
•_:•. SECOND HALF OF WORK SHIFT. MAY 1978
Range
PI ant b . ; NP/NTa _ jug/L
North 5/32 *0.8 - 2.0
South 2/29 <0.8 - 1.4
= Number of samples containing the compound; NT = Total number of samples.
bNo significant difference between results from the two plants.
77
-------
The results of the analysis of urine specimens collected in June 1978
divided according to whether collected during the first half of the work
shift or second half of the work shift are presented in Table 3-5 for HEX-BCH
and Table 36 for HEX. A number, of /the urine;samples obtained early in the
work shift"had detectable amounts of both H€X-BCH and HEX; In the case of
North Plant samples some exposure could have occurred prior to obtaining
the specimens. The HEX-BCH concentrations for the South Plant samples had
one value of 12>ig/L which was higher than the other samples. The values for
the other nine positive samples ranged from 0.4 to 3.5./ig/L, while the values
for the North Plant sample ranged from 0.4 to 7*5>ug/L. End of the work
shift samples (Table 35) indicated that urinary excretion of HEX-BCH increased
during the work shift for individuals at the North Plant (p^ 0.0001). HEX-BCH
excretion decreased during the work shift at the Maxson Plant and HEX excretion
did not change significantly at either plant. Comparison of the results
collected during the second half of the work shift in the May and June screening
indicated that at the North Plant excretions of HEX-BCH were higher in May
than in June (p = <0.010) and excretions of HEX were not significantly dif-
ferent (p = 0.051). Thirty-three percent of the early in the shift samples
had detectable concentrations of HEX-BCH. This increased to 76 percent for
the end of shift samples and the mean for these was 4.1/ig/L.
The prevalence of self-reported illness symptoms during the May and
June surveys, by plant and by shift, are presented in Table 37. The two
symptoms of eye irritation or burning watery eyes; and fatigue or malaise
were positively related to concentrations of HEX-BCH (p = 0.002 and 0.01,
respectively). For the May and June surveys there were also a number of
Siiqnificant.statistical relationships among prevalence of various symptoms
and plant, shift worked, age, sex, race, date of-. specimen, distance from
.North Plant and plant-shift interaction. The average number of symptoms per
worker was 1.26 at: the; North; pi ant; and 0.10 at ;the^Maxson Plant (p X 0.003).
The individual symptoms: headache, dizziness/"light headedriess, vomiting/dry
heaves and sleeplessness were more frequently reported in North Plant interviews.
Interviews conducted during the day shift revealed an average of 0.43 symptoms;
afternoon shift interviews, 1.50, and night shift interviews, 3.30 : (p =
0.0007). The average number of symptoms per worker, by shift and by plant, are
shown in Table 38. The plant-shift interaction symptoms was significant (p -
0.04). A plant-shift interaction was also found for the individual symptoms
headaches (p = 0.3), dizziness and light headness"(p = .0008) and tingling/
numbness of mouth and throat (p =0.0001). These symptoms were reported in-
frequently or not at all at the Maxson Plant. While at the North Plant they
were reported more frequently during the afternoon shift than during the day
shift and for two of the symptoms they were reported most frequently during the
night shift (Table 37). For 9 of the 16 symptoms there was a significant
difference between shifts in the reported rates.
Presence of fatigue, malaise was positively correlated with age (p =
0.003). In sixteen percent of the interviews with male participants, nausea,
lack of appetite, upset stomach or diarrhea were reported while none were
reported in interviews with females (p = 0.03).
The average number of total symptoms per worker was lower for black
.workers 0.29, than for white workers, 1.07 (p = 0.01). There were fewer re-
ports of the three individual symptoms of eye irritation, burning or watery
78
-------
TABLE 35. HEXACHLOROBICYCLOHEPTAOIENE (HEX-BCH) CONCENTRATIONS IN URINE
SAMPLES OF MEMPHIS WASTEWATER TREATMENT PLANT EMPLOYEES, JUNE 1978
Plant
North
North
Maxson
Maxson
Collection
Time
1st half
shift9
2nd half
shifta>b
1st half
shift
2nd half
shiftb
Total
Samples
54
54
23
19
Range
^g/L
<0.3-7.5
<.0.3-10.3
^TO.3-12
<0.3-1.6
Urine
Below
Detection
36
13
14
16
Concentration,>ug/L
less than
or equal
to 3
14
19
7
3
greater
than 3
4
22
2
0
a. 2nd half shift samples greater than 1st half samples (pug/L
<0.8-3.7
•CO. 8-3. 9
<0.8-4.8
<0.8-3.9
aNP » Number of samples containing HEX; NT = Total number of samples.
bThere were no statistically significant differences between plants or between
1st anHnd naif shift samples at eicner plant.
79
-------
TABLE 37. PREVALENCE OF SELF-REPORTED SYMPTOM
DURING MAY AND JUNE SURVEYS
May
Shift 1 Shift
Symptoms da* aft" ^htj day aft.
Number of interviews 36 3 3 I 23 5
(number of workers) (36) (3) (3) (23) (5)
1. Eye irritation, iff) lff?l(l?'d
burning watery
eyes
2. Fatigue, malaise 1051
a,b,c a,b,c a.b.c
3. Headache 1(1) 1(1) 2(1) '
4. Cough 1(1) i '
a :
5. Chest discomfort 1(1)
a,byc
6. Dizziness, light- 1(1) :.
headedness
a
7. Wheezing, short- 1(1) j
ness of breath i
difficulty in
breathing i
8. Fainting
a,d a,d ;
9. Nausea, lack of 1(1) 1(1)
appetite upset
stomach, diarrhea
a a a
10. Vomiting, dry 1(1) 1(1) 1(1)
heaves
a a
11. Irritation of 1(1) 1(1)
upper respitory,
sore throat
12. Gagging 1(1)
a,d
13. Tingling/numbness 1(1)
of mouth & throat
14. Loss of memory ;
15. Irritability [
a,b :
16. Sleeplessness l(i) ,
: " June _— .
— — North^Plant ; ^^ftfT^I^-
night day aft. night off day aft^J
-------
TABLE 38. AVERAGE NUMBER OF SYMPTOMS PER WORKER
PLANT AND SHIFT, MAY-JUNE 1978
Shift
Day Afternoon Night Off-Work
North Plant 0.63 2.43 3.44 2.33
(number of workers) , (54) " (7). (9) (3)
Maxson Plant 0.09 0.20 0.00
-(-number of workers) (33) (5) (1) (0)
81
-------
eyes; fatigue, malaise; and tingling/numbness of the mouth and throat, from in-
terviews with black workers than with white workers.
A higher rate of symptom prevalence was reported in June than in May (P =
0.0053). However, this maybe due to the May survey of illnesses being con-
ducted one month later. The prevalence difference also, existed for the
individual symptoms of eye irritation, burning watery eyes (p = 0.04), fatigue*
malaise (p = 0.008), and nausea, lack of appetite, upset stomach, diarrhea (P "
0.005). There was a higher reported rate of the symptoms of headache, cougn,
vomiting and dry/heaves, gagging and sleeplessness for people living farther
from the North Plant than among those living closer.
During July 1978 an additional 62 urine specimens were collected from
several groups in Memphis, totalling 40 people. These groups consisted of
North and Maxson plant employees, a crew of city employees sampling a sewer
from the pesticide manufacturer and an additional control group not involved
with either treatment plant. Results of analyses of these specimens are
presented in Table 39. Six specimens were collected from four North Plant
workers during a period of intense chemical odor. Two of the workers re-
ported symptoms of dizziness, headache and drowsiness. Urine from one of
these two workers contained 2.0,/jg/L HEX-BCH three hours into the work shift
and urine from the other contained 2.9jug/L HEX at the end of the work shin-
Only one other urine specimen of those evaluated in July 1978 contained a
detectable amount of HEX. The sample was an end-of-shift specimen during
the same period. HEX-BCH was detected in urine from four of the other 20
specimens collected from Noth Plant workers during July 1978 and in five of
^0 specimens collected from the sewer sampling crew. One of the seven urine
specimens collected from Maxson Plant workers and one of the nine from the
Crve£ru°ntT?ls from e1sewhere in Memphis contained detectable amounts of
Htx-BLH. The positive sample from the Maxson Plant worker was collected at
the start of the work shift, an end-of-shift sample from the worker did not
S?5 S™ dete^able/oncentrations of HEX-BCH. The other control worker with
HEX-BCH was the wife of an agricultural chemical company employee. Combining
Jh! ^-SKi/T the North Plant workers and the sampling crew and comparing
them with the Maxson Plant workers and the other controls, the results from
the former groups are significantly higher (Table 40).
cond.JtPri^nVT eHted1l5 Tab1e 41 9ives the results of a urine screening
rniWMn ! Se?Jember 1978 at the time of the regular quarterly specimen
had dPtPrSJT he sei;oeP1def"i°l°9ic study. One individual at the North Pljf1
had detectable concentration of urinary HEX-BCH. HEX was not found in any
the North Plant urine samples and neither HEX nor HEX-BCH was found in tne
samples of individuals from the Maxson Plant or from the Mill Creek Plant "
Cincinnati, Ohio. However, HEX-BCH was detected in the range of 0.3-l.O^jj'1
G&W
, e range ..
°f ft MtS&W ^^^ tested. An ^sample at MLG&W
ivelv Thv i>J9^m Of HEX'BCH and heptachlorobicycloheptene, respect
views'arl shown {iT^^Wi!"15 rep°rted dur1n9 the September^ 1978 inter-
re urn9 e eptemer
wor woA The avera9e number of symptoms reported per
(Cincinnati^ n n? T^ °<29; MaXS°n P1ant' °'48' MLG&W> °'16 a"d Mi11 ?"
0 48 f?SS liv ^l\ 56 Iatf f°r the Maxson Plant ^creased from 0.08 to
0 49 to 0 29 September while that for the North Plant decreased fr*
82
-------
TABLE 39. RESULTS OF ANALYSES URINE COLLECTED FROM VARIOUS
GROUPS IN MEMPHIS, TENNESSEE DURING JULY 1978
HEX
Group
North Plant Worked
N°rth Plant Workers
Sampling Crew
Maxson Plant Workers
Otter Controls
Np/NTa
2/6(2/4)
0/20(0/12)
0/20(0/11)
0/7(0/5)
0/9(0/8)
Rangeb
tug/L)
1.5-2.9
zO.8
xiO.8
^0.8
±0.8
HEX-BCH
NP/NTa
3/6(3/4)
4/20(4/12)
5/20(4/11)
l/7
-------
TABLE 40.. COMPARISON OF RESULTS OF HEX-BCH ANALYSES OF URINE
SPECIMENS DURING JULY 1978 FROM NORTH PLANT WORKERS..AND
SAMPLING CREW WITH THOSE FROM MAXSON PLANT WORKERS
AND OTHER CONTROL WORKERS
Urine Concentration/jg/L
Worker Group
Total
Workers
Range
Below
Detection
Less than
or Equal
to 3
Greater
than 3
Combined North
plant and
sampling crew
27 /.. 8-4.0
16
Maxson Plant
and other
controls
13
. 8-1.3
11
TABLE 41. URINE SAMPLES OF WASTEWATER TREATMENT PLANT EMPLOYEES
COLLECTED AT THE END OF SHIFT IN SEPTEMBER 1978
Plant
North
Maxson
MLG&W
Mill Creek
Cincinnati , Ohiob
Np/Nra
0/55
0/32
0/29
0/20
HEX
Range
<0.8
^0.8
<0.8
<0-8
HEX-BCH
Np/N
1/55
0/32
7/29
0/20
Range
'<0.3-0.8
<0.3
<0. 3-1.0
Np - Number of samples containing the compound; NT - Total number of
samples. ..
DCollected in October 1978.
84
-------
TABLE 42. PREVALENCE OF SELF-REPORTED SYMPTOMS
Symptom
(Number of Workers)
1. Eye irritation, burn-
ing watery eyes
2. Fatigue, malaise
3. Headache
4. Cough
5. Chest discomfort.
6. Dizziness, light-
headedness
7. Wheezing, short-
ness of breath,
difficulty in
breathing
8. Fainting
9. Nausea, lack of ap-
. petite, upset sto-
mach, diarrhea
10. Vomiting, dry
heaves
11. Irritation of upper
respiratory system,
sore throat
12. Gagging
13. Tingling/numbness
of mouth & throat
14. Loss of memory
15. Irritability
16. Sleeplessness
17. Other
Plant
North Maxson MLG&W Mill Creek
45 31 34 15
2
2 1
! 4 2
1
1 2
1
V '*.'
42 1 !
2 J =
TOTAL 13 J5 | 0>07
AVERAGE RATE 0.29 0-^8 U-1D
85
-------
Summary of Results of Analysis of Urine Data and Illness Symptoms
Urine concentrations of HEX-BCH were significantly hiqher in North Plant
workers than in Maxson Plant workers in both May and June 1978, The col-
lection of specimens early and late in the work shift during June revealed an
increase in urinary excretion during the work shift among North Plant workers
lh °Surat the Maxson Plant' No Differences between
for HEX. Between the May-June collections and the
» H°iJvCtT ^er* was an overa11 decline in urine concentrations of
and HEX. It should be noted that worker awareness of the presence of
tnese^compounds increased considerably after the June collections as the
result of meetings and other communications -and they were advised to leave
work areas when the chemical odors were intense. ««Jvi>ea LU
f ithe K^,?]8"* reP°rted * hi9ner rate of illness symptoms
andK0une than dld Maxson P1«nt workers, The presence of eye
or burning, watery eyes and fatigue/malaise were positively
Unni concentrations of HEX-BCH. If the statistically significant
onrrholenCeS °I ^me °f the sWtoms detected are not due to chance
while'nnt St ^SU?SeStKthat When far enou9h awa* from ai^ contamination
cause thl JnS rAn ? ^°me Te suscePtible to them when at work. Be-
1 mits for I i ^ 5? ntr;tlo"s of, HEX and HEX-BCH were below detectable
limits for all and almost all workers, respectively in S^ntPmbPr it is not
'"beali groups
Environmental Monitoring
lecte^afthe £?h JX^S*1 9aVhromatograms of influent wastewater col
stSdv The North Han? ?IS?n ^eatf nt Pla"ts duHng the time of this
s 1n!ludin« 2?{ lr^ u C]ea[ y shows ^e' presence of several con
°r VaP°rS bef°re Slgn1f icant «P«ure to workers
c™berarenan appeancTioY^ 'f ttd,at the Wet wel 1 and grU
HEX HFX RPH hint, ki apP?ara"ce to the chromatograms of the wastewater. .
sa^l ^ atH't eP or 0nantyCl?h-Ptene 3nd Chlordene were p"esent in th£ &^
,,,,'S'!.S - ''™ - ''
, .t -s ''
86
-------
o
C£
O
15 MIN
NORTH PLANT
15 MIN,
MAXSON PLANT
Figure 4. Gas chromatograms of extracts of
influent wastewater samples.
87
-------
The concentrations of four chlorinated organics found in the influent
wastewater of the Memphis North Plant during the time of air sampling and
urine screening studies are given in Table 43. Although these values do not
represent the values for the complete month, they are indicative of the con-
centrations at various times. The concentration of HEX-BCH was generally
about 300 ppb, although much lower levels were measured during October and
November. Heptachlorobicycloheptene and chlordene concentrations seem to^be
more variable ranging from about 17 to 1980 ppb. One sample collected during
September had a chlordene concentration of 1980 ppb. The September samples
listed in Table 43 were collected at the time of a reported "spill" which
occurred after the urine screening that month. The analysis of wastewater
and air samples indicated that heptachlorobicycloheptene was at least one of
the major constitutents in the "spill."
Area air samples collected at various locations in the treatment plant
provided information about the magnitude of aerial dispersion of these semi-
volatile compounds. The concentrations of these substances found in air
samples collected in the wet well, a building with a ventilation system, and
the grit chamber, are presented in Table 44. The levels at the wet well
were generally higher than at the grit chamber. HEX was found at levels as
high as 39jug/m3. The highest levels of HEX-BCH and heptachlorobicyclo-
heptene were 280>jg/m3 and 200/ig/m3, respectively.
Personal air monitoring of 11 workers at the North Plant and 9 at the
Maxson Plant was carried out in September 1978. Eight-hour personal air
samples were collected during the work shift and during the workers' off-
snm time (home sample). The work shift sample for most of the North
Plant workers contained HEX, HEX-BCH, heptachlorobicycloheptene and chlordene
(Table 45). There was no obvious relationship between work area and the .
extent of exposure. However, for North Plant workers the air concentrations
for the four compounds found from personal air monitoring during the work
shift were in good agreement with the concentrations determined from area
air samples collected at the grit chamber. Several of the home samples of
North Plant workers also had detectable amounts of some of these compounds,
but the levels were considerably lower than the work shift sample. The
samples of the workers at the Maxson (South) Plant were below detection
limits except for one work shift sample which contained a trace of HEX-BCH.
All urine samples contained less than detectable concentration of HEX, HEX-
and heptachlorobicycloheptene.
Medical
Liver function tests were performed in November 1977 and in November
1978. No differences between values for workers at the North Plant, Maxson
Plant and MLG&W were detected for either year. Total serum bile acid de-...r.nt
terminations performed on the November 1978 serum did not reveal any sigmfic
differences between the groups.
DISCUSSION
Hexachlorocyclopentadiene (HEX) is a semi-volatile, highly reactive
chlorinated compound which is only slightly soluble in water. Information
-------
TABLE 43. CONCENTRATIONS OF SELECTED ORGANIC COMPOUNDS IN
INFLUENT WASTEWATER AT MEMPHIS NORTH TREATMENT PLANT, 1978
n Concentration, jmg/L
^il "HEX HEX-BCH HEX-VCI* Chlordene
June 3 334 57 87
Au9"St <0.8 329 115 216
September 4 292 668 58
\c0.8 11 17 32
*Heptachlorobicycloheptene
89
-------
TABLE 44. AREA AIR SAMPLES COLLECTED AT THE MEMPHIS
NORTH PLANT, 1978
A. WET WELL
Date
May
June
September
October
November
B. GRIT CHAMBER
Date
May
June
July
September
October
November
HEX*
^0.03
18
8
15
39
HEX
<0.03
6.3
^0.03
40.03
0.04
12
Concentration.
HEX-6CH i
219
278
25
2
7
Concentration,
HEX-BCH
4.1
65
0.5
0.5
1.2
2.6
juq/m3
HEX-VCt*
87
15
200
1
85
vuq/m3
'HEX-VCL
1.9
1.5
0.7
1.1
1.0
4.3
Chlordene
45
16
44
0.1
7.8
.
"TFTofainf
0.9
5.3
2.3
2.7
0.8
=^=====^
threshold Limit Value for eight hour sample is 110>ug/m3 (10 ppb).
**Heptachlorobicycloheptene
90
-------
TABLE 45. PERSONAL AIR MONITORING OF WORKERS AT THE
MEMPHIS NORTH WASTEWATER TREATMENT PLANT
Concentration,^ig/m3
Worker
1
2
3
4
5
6
7
8
9
10
:==r:
HEX
Wa Ha
0.1 " — b
NSC
0.31
NS
«'
NS
0.42
NS
/•--..;.. NS
NS
0.08
HEX-BCH
W H
0.12
0.02
0.40
0.46
0.05
0.22
0.33
0.37
0.24
0.48
0.10
0.02
NS
—
NS
NS
0.03
NS
NS
NS
--
HEX-VCLa
W H
0.09
--
0.16
0.31
0.02
0.10
0.21
0.21
. • — : , • .
0,22
0.02
0.04
NS
—
NS
—
NS
0. 10
NS
NS
NS
Chlordene
W H
0.20
—
0.21
1.25
—
3.32
1.19
.-- .
0.94
—
__
NS
»
NS
• «
NS
—
NS
NS
NS
--
a
W - designates the work shift sample, H - designates the home sample.
(•-)
NS indicates that no sample was obtained.
indicates the sample was below detection limit.
Heptachlorobicycloheptene.
91
-------
concerning human health effects of exposure to HEX is very limited. What
oilH nr OS" ab°UH ^Ute human toxic1t* 1s based largely upon isolated
nfhn 2 accidents involving pesticide workers, laboratory workers or
SeriencedaVi>?i?$HEnatlfnta; C°ntact ?th HEX' ^trial workers have ex-
perienced irritation of the eyes and upper respiratory tract and headaches
upon exposure to HEX vapors; and'burns upon contact with liquid1 (73-75).
a Sinate^!eH(76) jnve^ifted the absorption, metabolism and excretion of
(ma e9ratO A of radiolabelled HEX ( JC.HEX) in laboratory animals
urine duH™ >£ffOX1?a^ 29% °f tota1 C dose was eliminated in the .
9 fe flrs^ 24 hours after administration. Other routes of elim-
were feces and probably the respiratory tract. The metaboic pathway
5Ut-the Mehendale "tudy suggests the possibnity of
exposed to HExhvh f;00109^'1^^0"15 have beerl observed in mammals
y alaton' dermal and oral routes of administration
1on9 term exposures may
hat E BCH
'
Ot?er compounds but there are indication
that thelLV FV RrM°X1Ci?SKHEX (68)' Hypothetical ly, if it is assumed
tnat tne ILV for HEX-BCH would be equal to the TLV for HEX MOO Wm3 or 10
w d barCea°nS lS the Wet ^1 duri ' ^^une 1978
exposure levels
single
may not adequate! /reflect the actual
1
vapor!. dl
chemical contaminants as aerosols
92
-------
is clear from the data collected during the study that these compounds
not present at the Maxson Plant at the same levels as at the North Plant.
'He small quantities of HEX-BCH detected in the urine of some MLG&W employees
could have been due to the proximity of MLG&W to the pesticide manufacturer
°r to ambient levels at the sites of their work that day away from their
central facility.
.. ^At the present time there is inadequate information available regarding
tneTiealth affects of chronic exposure to HEX and related chlorinated
compounds. Epidemiologic studies of workers who are occupationally ex-
Posed to HEX are lacking and there is no information available concerning
the potential health effects resulting from chronic environmental exposure
to HEX via contaminated air and/or water. The Louisville, Kentucky ep-
isode (64,65) demonstrates a case of acute exposure to toxic contaminants
'i"om wastewater. The situation described in this report is a case of
essentially continuous chronic exposures with intermittent acute exposures.
unfortunately, only limited information is available on the toxicity and
wealth effects of these chlorinated organics. Thus, the significance of
urinary excretion of these substances and the possible relationship to
"ealth effects is not known.
93
-------
SECTION 7
VIRUSES IN *i A MAS™ATER
The purpose of this project was to collect, enumerate, and identify
viruses in aerosols at a spray irrigation site, the Muskegon County, Michigan,
Wastewater Management System Number 1, and to examine wastewater samples for
the presence of coliphage and animal viruses in order to compare airborne
levels to the levels in the wastewater. The presence of animal viruses in
aerosols would indicate a potential health risk for treatment plant workers.
Standard Plate Count bacterial analyses were also perf armed for comparison
purposes. The sampler used in this study was the Army prototype XM2 Bio-
logical Sampler/Collector.
SITE DESCRIPTION
The Muskegon County Wastewater Management System Number 1 was a lagoon-
impoundment, spray irrigation treatment facility which serviced 13 munici-
palities and five major industries (77,78). The system served a population
of approximately 180,000. The average daily flow of wastewater was 106,000
M3 (28 million gallons). Approxmately 50% of this flow came from a paper
mill and another 5% from chemical manufacturers. The system consisted of a
4,450 hectare (11,000 acre) site which contained three aerated biological
treatment cells (aeration basins), two storage lagoons of 345 hectares (85U
acres) each with a total storage capacity of 19.3 million cubic meters (5-1
billion gallons); and 2,270 hectares (5500 acres) of land irrigated by 54
center-pivot irrigation rigs (Figure 5).
Raw waste entered aeration basins number 2 and 3 which were operated in
series. Aeration basin #1 was not in use during this study. Each aeration
basin had 3.2 hectares (8 acres) of surface area and was equipped with 12
mechanical surface aerators and 6 mixers. A settling pond and an
outlet lagoon, located adjacent to the aeration basins, were also not in use
during the study. After an average detention time of three days, the efflu~
ent flowed to the storage lagoons. The lagoons provided additional time for
stabilization and settling of wastes and for storage of wastewater during
the fall and winter seasons when irrigation was not practiced. The waste-
water was not disinfected. Wastewater flowed from the lagoons via open
channels to the two pump stations for distribution to the 54 irrigation
rigs. These rigs, equipped with downward-directed aerated high pressure
spray nozzles, have a radius of 229 to 396 meters (750 to 1300 feet) and a
rotation rate that varied from 1 to 7 days. The wastewater application rate
was approxmately 5700 M3/day/rig during the growing season. An underdrainage
94
-------
TO K
TO KOKA LAKE
Figure 5. Muskegon County Wastewater Management System no. 1.
95
-------
system collected leachate from the irrigated fields for discharge into nearby
waterways .
METHODS
XM2 Biological Sampler /Collector
The Army prototype XM2 Biological Sampler/Collector, oh loan from the
Edgewood Arsenal, was manufactured by the Environmental Research Corporation.
This sampler/collector removed airborne particles from the air flow, and
concentrated (in an 8:1 ratio) classified particles (2-12 m) in a smaller
volume of air before moving them into a collection liquid via a combination
of impingement and scrubbing. This apparatus could store discrete 10 ml
portions of the spent collection liquid in a temperature-controlled en-
vironment or in a large reservoir, depending on which option was chosen. A
schematic of the sampler air unit is shown in Figure 6. The sampler pro-
cessed 1050 liters of air per minute and could run continuously for periods
of time up to 24 hours without the addition of more collecting liquid (79).
Sampling liquid was collected at a rate of approximately 2.0 ml per minute.
A schematic of the sample gas flow through the XM2 Sampler/Collector is
shown in Figure 7.
Collection Liquids
le "Jlection liquids used were Earle's Lactalbumin Hydrolysate
-ann510t^S' ELH without antibiotics, and distilled water. Samples
^U J? J an^biotics were assayed for viruses. Samples col-
u LH Wlthout antibiotics and distilled water were assayed for bacteria
coiiprvage.
de1onl7PriWrf5c?J?i HP U+ing 102 S 10x ELH 'Stock. (GIBCO), 870 ml sterile
Wpr2 JJoJn }l H ied Water' a2?,30 m1 7'5% NaHC°3 stock! All stock solutions
STloilS1ii"d.di!e8:eSSUre flltrati°n- Whe" ^tibiotics were required, the
, ' 3 soc s
flltrati°n- Whe" ^tibiotics were required,
1>0 m1 p!nV-!!-n"rtreKmrin stock> "Gaining 100 Units/ml of
n 9 mi £ Cl1hn.G and 10° u9/m1 Dihydro-streptomycin
0.2 ml Amphotericin B stock (1 jug/ml)
0.5 ml Tetracyline stock (12.5/jg/ml).
Measurement of the Air Velocity of the Sampler Air Unit
DrohPT?vnf1^l0Ya^ throu9h,tne sampler air unit was checked using a
Sboarfro I," S«"^-mSter,(M°de1 441 ' Sierra Instruments, Inc.). A
to th! «mSii • . ,19h 3nd 4'8" in d1amet^ was constructed and attached
an oSen?naP n th 'n'^i Readin9S were taken ^ inserting the probe into
iStSK Thl fn ° I3" °Cated °ne f1fth tne Co11ar ^ngth above the air
see FiaurS R! readin9s' ^aken at different locations within the collar.
obtain til a?i'u!?re-;Vera?fd and multip1ied by the cross-sectional area to
the air SSinMtw 1? J J' • ^ *^}er air velocity was within + 10-15* of
the air velocity stated in the Operator's manual (1050 liters/minute) at the
96
-------
Figure 6. XM2 sampler air unit.
97
-------
IO5O UT»M SAMPLE
IM
AMO
S*PA»ATION AND
C0NCCKITAATIAN
PARTICLES
10
09
f
/
1
S-
<,
L{
III
I
IOOO LPM
|7zsu««-
r:
iL*
••• «•«!<
1
I5UPH
FLOW
j—
1
(
•
"* * "
VJKT
AMD AinV
LIQUID
1
»-
FL«vl
OM
|«:
1
HO
WM
FMM
SCPM
PLOW
AIR PLOvl WITH
. PA^TI£LKft<£^.
J675LPM-HIOLFM)
/
y.. 50 UPM SAMPLE GAS FLOW WITH A14.
/^ "PARTicuss OF SAMT»UE ^AS > 12.^*.
30 LTT1
/ \\ C.A»OUT
^-
-------
Area =» "
^cation
Nuraber
1
2
I
Area
Distance Prom Probe
Openlng(X)
11/32*
1 9/16"
I 7/16"
r = PI + r2
r » 2.U* or 0.2'
r, = 1.7"
r = 0 7"
* n V i (
Area = trr^
Area = T<(0.2 ft.j2
Area = 0.126 ft.2
Figure 8. Cross-section of the air collar of the sampler
showing the location of the air flow readings.
99
-------
settings recommended by the Edgewood Arsenal personnel. Variation in reading
were due primarily to fluctuations in the blower motor speed.
Cleaning and Sterilization Procedures
The sampler tubing was sterilized by fining it with 100-200 mg/1 Clorox
solution, prepared in sterile distilled water, and allowing it to sit over-
night. After flushing the tubing with freshly prepared Clorox solution, the
tubing ends were immersed in sterile containers containing 100-200 mg/1
Clorox solution. After a 1-hour contact time, the tubing ends were rinsed
with sterile distilled water, and the tubing was flushed with sterile dis-
tilled water until a negative Clg spot test using DPD#1 Powder (LaMotte
Chemical Co.) was obtained. Aliquots of the upper and Tower distilled water
rinses were then collected in sterile containers to check for tubing steril-
ity. These distilled water rinses were checked for the presence of bacteria,
fungi, viruses, and coliphage. If any were found, the cleaning procedure
was repeated. Sterile air was drawn into the lower tubing via a sterile
trap system (see Figure 9). Sterile distilled water was left in the upper
tubing until the sampling day. The ends of the tubing were given a final
1-hour contact time in Clorox solution, rinsed with sterile distilled water,
and covered with sterile foil.
The sampler air unit was sterilized immediately after every run and
again after cleaning. The air unit, wrapped in brown kraft paper, was ex-
posed to ethylene oxide gas (12% by weight) for 4 hours in an Amsco Cryotnen >
with 30-50% humidity. The air unit was then aerated for 2 hours in an Amsco
Ethylene Oxide Gas Aerator. The air unit shroud was sterilized similarly.
The wet collector of the air unit was cleaned after every sampling run, ana
the entire air unit was dismantled for cleaning every other run. Sterility
of the air unit and shroud was checked by ethylene oxide sensitive tape ana
by the use of Bacillus subtil is, var. niger spore strips (Raven Biological
Labs, Inc.), taped inside the air unit and shroud prior to sterilization.
These were removed at the sampling site during sampler assembly, placed in
the envelope with the positive control, and inoculated into thioglycollate
broth upon our return to Cincinnati.
Sterility Tests
The distilled water rinses from the sampler tubing were checked for the
presence of bacteria by inoculating several ml of each into Trypticase Soy
Broth (TSB) tubes and incubating at 37°C for one week. The tubing was con-
sidered free of bacteria if there was no growth in TSB at the end of that
time. At least 2 ml of each rinse were put on the surface of Trypticase boy
. ur
Agar (TSA) plates (1 ml/plate), and the inoculum was spread by sterile n°J^
sticks. The TSA plates were observed for fungal growth after room temperatui
incubation for 1 week. Aliquots of both distilled water rinses were assaye
for viruses and coliphage by the usual procedure, described elsewhere.
The air unit, shroud, and control spore strip envelopes were cut with
scissors that had been dipped in ethanol and flamed. The spore strips were
poured aseptically into tubes of fluid thioglycollate that had been heated
to drive off any absorbed oxygen and then cooled. The tubes of broth were
100
-------
AIR
IOO-20Q PPM CLOROK
ABSORBEMT CCfTTCH
9. Sterile trap system for introduciog air Into lower
tubing of the sampler.
101
-------
incubated for one week at 37°C and checked for growth daily. Growth in the
positive control tube and absence of growth in the other two indicated that tne
air unit and shroud were sterile. The air unit and shroud were found to oe
sterile in all runs.
Heathkit Portable weather Station, Model ID-1290
This portable weather station was used to monitor weather parameters
(temperature, relative humidity, barometric pressure, wind direction ana
velocity) during all air sampling runs. Readings were generally taken
10-15 minutes. Cloud cover and other general weather conditions were
recorded.
*
Sampling Procedure
Samples were collected during six sampling trips from April to October.
1979. All aerosol samples were collected with the XM2 Biological Sampler/
Collector at aeration basin #2, downwind of an operating surface aerato
whenever possible. Figures 10 and 11 show the location of the sampling site s.
These sites were upwind of the aeration basin when the wind was from the west
or northwest.
The sampler was placed about 1 meter from the aeration basin wall
(about 0.6 meters high) which was 2.5-3 meters from the edge of the basin w
water. The wastewater surface was usually about 2 meters lower than the
of the wall. The sampler air intake was located above wall height, 0.7 ®
from the ground or about 2.7 meters above the basin wastewater surface
Figure 12). The sampler was located 20-25 meters from the center of the neares
operating surface aerator on all sampling dates, except 5/15/79, when it was
located approximately 90 minutes down the road between aeration basin #1 a"
#2. In that position, it was 70-73 meters from the center of the neares
operating aerator.
After the weather station was set up at the sampling site the XM2 air unit
was unwrapped and placed in the sampler. The sterile tubing ends wer
connected to the wet collector liquid inlet and outlet ports. Sampler hours
were recorded, and the airflow settings were checked before the '
,
collecting liquid was pumped into the tubing. The sampler case was
closed, and the shroud was put into place at the air inlet. Two 2-hour s
were collected; the first in ELH without antibiotics, and the second m tun
19, 1979, an additional sample was collected J'
r
with antibiotics. On June 19, 1979, an additional sample was colec
distilled water prior to collecting the two samples in ELH. Weather parameter
were monitored during .all sampling runs.
As soon as possible after sampling, water samples were collected at the
aeration basin manhole (raw influent), the west lagoon, and the ditch at in
north pumping station, before and after the location where the liquid nitroge
was added to the wastewater. In addition, foam that accumulated on the surraj
of the aeration basin was repeatedly scooped into a sterile beaker and ali°w*,
to settle. When a sufficient quantity of settled foam was obtained, a smai
quantity of ELH without antibiotics, distilled water, or 10% beef extract, pn
7.0, prepared in Mcllvaine's citric acid-sodium phosphate buffer (80), was
102
-------
AERATION BASINS
o
oo
"t
WEST
STORAGE
LAGOON
A, MAY 15,1*17*? ONUY
* JUNE 19,1979 ONLY
n JULY n,i
-------
mn
\
WEATHER
STATION-
CONSOLE
WEATHER
STATION_
ANENOMETER
AERATION
BASIN *1
(NOT- IN
OPERATION)
.SCALE
0 10 20
y
^x
1
• !'•• ^
m*m+m
BER c
ING
6/19/7'
7/17/79
LO/2/79*
/21/79.
5/15/79
ROAD
WAD
t
^ ^-^^
,V^ N
( £!*£> OPERATOR
^*> AERATOR
AERATION BASIN £
» AERATOR NOT
IN OPERATION
^OA
/T '0? OHEKATING
A Cr^tf* AERATOR
WEST-LAGOON
Figure 11. Schematic of aeration basins #1 and #2
showing the location of the XM2 biological
sampler/collector during the specified days
of the sampling runs.
104
-------
AIR
SAMPLER WALL
J7 r-
0. 7
METEKS
•£, n
,
_ 0.6
METERS
KUAU
SCALE
0 I
METERS
AERATION BASIN #2
2.5-3 METERS
\ WASTEWATER
\
V
Figure 12. Schematic showing the position of the XM2
biological sampler/collector in relation
to the edge of the wastewater.
105
-------
added. All samples were iced immediately and frozen until assayed at tn
University of Cincinnati. Sediments of deposited foam, found on the slde.?
the aeration basin and on the road adjacent to it, were collected in steri
beakers using sterile spatulae and forceps. The samples were iced and stored a
10°-12° until assayed.
Sample Processing and Virus Assay
Qfi
One hundred-mi water and 50-ml foam samples were centrifuged. for
minutes at 1800-2000 rpm in an International Centrifuge, Size 2, ancIt«
supernatant liquid was syringe-filtered through a 10% beef extract (0*° .-iii.
Lemco Powder), pH 7.0-pretreated O.AB^/um and 1.2/jnv membrane filter (Mi" "
pore) sandwich in a Millipore Swinnex 47-mm filter holder. The beef extra
was prepared in Mcllvaine's citric acid-dibasic sodium phosphate buf
7.0 (7-4). The filters were then eluted with 5 ml of the beef extract,
to ten ml of the beef extract was added to the centrifuged sediments an
with a Vortex mixer. The mixture was allowed to stand overnight at ,
before centrifuging for 30 minutes at 1800-2000 rpm. The supernatant I1>e*'
sediment eluate) was syringe-filtered through a 10% beef extract, pH 7'0~p!V
treated O.AS^m membrane filter. In addition, air samples in ELH with ant
biotics were syringe-filtered and the filters eluted by the same met"
described for the water and foam samples.
Five-mi aliquots of the water and air sample filtrates and 0.5-ml
of the filter eluates, sediment eluates, and foam filtrates were ir.-- .
into 6-ounce prescription bottles containing the continuous line of BGM cei | »
derived from primary African green monkey kidney cells. After an incubati •
period of two hours at 37°C, the bottles were overlaid using the pjjg.
technique of Dulbecco and Vogt (81), as modified by Hsiung and Melnick (oc->'
In addition, aeration basin raw influent samples were processed by
second method, a modification of the low pH membrane filter adsorption-elUj-1
method (83), as previously described (80,84). Samples were clariflf.ni.
pressure filtration through a 127-mm Gelman Metrigard prefilter and a ^1'
pore 0.22-xim membrane filter in a Millipore 142-mm stainless steel _fi '*
holder. The filter and prefilter had been pretreated with 500 ml of 0.1* TWLe
80 and washed with 2 liters of sterile distilled water. After eluting *
clarifying filter and prefilter with 3% beef extract (40 ml/liter of samp
clarified) and collecting the eluate by pressure filtration, the filt;£r rer
prefilter were aseptically removed for a second elution in a sterile bea*
with enough 3% beef extract to cover. The 3% beef extract had been
in
with enough 3% beef extract to cover. The 3% beef extract had been preparea
Mcllvaine's citric acid-dibasic sodium phosphate buffer, pH 7.0 (0.12 g ci"\
acid, 1.34 g NaHP04.7H20, and 3.0 g beef extract in 100 ml distilled watery
The second eluate was poured off and centrifuged for 30 minutes at 180lr>e)
'
The second eluate was poured off and centrifuged for 30 minutes at 180
rpm prior to 'iltration through a 0.22-and 1.2-jum membrane filter (MiU
sandwich, .contained in a Swinnex 47-mm membrane filter holder. These eiuat-
were designated clarifying filter eluates one and-two. MgCl? (I-2 9/11tSr-aS
clarified sample) was added to the clarified wastewater sample, and the pH JJ«
adjusted to 3 with concentrated HC1. The sample was then pressure fiJ^JJJ
through a second 0.22->um Millipore filter in a 142-mm holder. After elu* Jg
the viruses from this membrane filter for 30 minutes with 3% beef extract I*
ml/liter of clarified sample), the eluate was forced through the filter by a
106
-------
pressure and collected in a sterile container. The acid filtrate was
neutralized to pH 7.0 with NaOH, and a portion (100 ml) was later assayed along
with the concentrated sample viruses and clarifying filter eluates 1 and 2.
The concentrating membrane filter was then eluted a second time for 30 minutes
in a sterile beaker with enough 3% beef extnact to cover. The eluate was poured
off and filtered through a 0.22-um membrane filter in a Swinnex 47-mm filter
holder. Five-mi aliquots of the neutralized acid filtrate and 1-ml aliquots of
the concentrated sample viruses, concentrating filter eluate, and clarifying
filter eluates were inoculated into 6-ounce prescription bottles, containing
BGM cells, and assayed as .previously described.
Well-isolated plaques were picked from tissue culture bottles inoculated
with the July 17, 1979 raw influent sample, processed by the membrane filter
adsorption-elution method. After the picked plaques were placed in ELH and
inoculated into BGM tube cultures, the cultures were incubated at 37°G for up
to 10 days. The cultures were checked daily for cytopathic effect (CPE), and
those exhibiting 75% or greater CPE were stored at -70°C until further passage
and identification could be done by the Clinical Virology Laboratory in the
University of.Cincinnati College of Medicine. The enterovirus isolates were
typed with Lim-Benyesh-Melnick serum pools.
Sediments of deposited foam, collected from the side of the aeration basin
and the road adjacent to :it, were weighed and placed in sterile beakers. For
every 15 g of solids, 40ml of 10% beef extract, prepared in Mcllvaine's citric
acid-dibasic sodium phosphate buffer, pH 7.0, was added to produce a slurry.
More extract was added if necessary to allow proper-mixing (79). The slurries
were mixed for 30 minutes on a magnetic stirrer and centrifuged at 17,100 x
g in a Sorvall SS-3 Automatic'Superspeed Centrifuge for 30 minutes. The
supernatant* were filtered through a 0.45-and 1.2-jum membrane filter sandwich
in a Swinnex:47-p membrane filterholder. The filtrates (i.e^, foam sediment
eluates) were assayed for viruses, 0.5 ml per bottle, in the usual manner.
Coliphage Assay
Untreated air, water, and foam samples were assayed by the soft agar
overlay method (85), using two E. coli hosts (ATCC numbers 13706 and 11303).
Sample dilutions were made in O.D5 M potassium phosphate buffer (5.17 g KHpPOA
and 9.56 g K£HP04 in 2 liters distilled water). Assays using a third E. coli
host, ATCC number 15597, were also done using the soft agar over lay "method
described by Safferman and Morris (84). Centrifuged, nonfiltered foam
sediment eluates were assayed for coliphage by the same methods. In addition,
a portion (*-170 ml) of each air sample was syringe-filtered through a 10% beef
extract, pO.O-pretreated 0.45-dum membrane filter before assaying by the Kott
MPN method (86).
Bacterial Assay
Using Standard Methods Agar, Standard Plate Counts, as described in
Standard Methods for the Examination of Water and wastewater, 14th edition
(30), were done on all water and foam samples, as well as the air samples
collected in ELH without antibiotics.
107
-------
RESULTS AND 'DISCUSSION
No animal viruses were detected in the -air samples c°11efr*ed Animal
Muskegon County Wastewater Management System Number 1 .(Table 46). * •
viruses, found in the water samples (Table 46), decreased as the wast.ew°di-
progressed through the facility, due to aeration, detention, ana
mentation. Because no viruses were detected in the pump station water j ^
prior to distribution by the irrigation rigs, no air samples were collet ^
the fields near the rigs. Fourteen viral isolates picked from the July '/> cflX.
raw influent sample were identified as follows: two Poliovirus 3, nin h
sackievirus B2, and 3 echovirus 7. One isolate contained an eriterovirus.
was riot typable with the pools, and two did not contain enteroviruses.
ipc
Using the centrifugation-filtration method, all raw influent sample
found to contain viruses, ranging from 50 to 400 plaque-forming units
(PFU) per liter (Table 47). Most of the viruses recovered were present •
sample supernatant liquids. Viruses were found in the sediment eluates
two of the samples. No viruses were detected in the filter eluates. i" Of
out of four samples, the centrifugation method recovered higher |m°" ad-
virus in the supernatants than in those done by the membrane f,
sorption-elution method. Since the centrifugation-filtration method
within one week of sample collection, there was less chance of virus a
to wastewater solids than in the other method, done at a later date
Using the membrane filter adsorption-elution method, viruses f°"nc* o
from 0 to 456 PFU per liter (Table 47). When the filters were eluted, mj£j.ane
the viruses in the raw influent samples were found adsorbed to the mem ^e
filter and/or the particulates separated out during clarification °ters
samples. Lack of sample prevented a full-scale examination of all Par les by
involved in the concentration of the viruses in these wastewater samp'
this method.
Aeration basin surface foam and aeration basin and road foam seen
eluates were found to be toxic to the BGM cells. An attempt to seParat
e
toxic material in the foam, by diluting the sample 1:100 with distil \Q
water and using the membrane filter adsorption-elution concentration P Of
for viruses, proved unsuccessful. Therefore, the presence or absen
animal viruses in foam and foam sediments could not be determined.
h
Although no animal viruses were found, coliphage, present in muc n
quantities in the wastewater, were recovered in the air sampled ^,706
aeration basins, using three different £. coli hosts (ATCC numbers. ' ^
15597, and 11303). Coliphage levels rangedTrom 0-9 per cubic meter °i
sampled (Table 48).
No coliphage of any type were recovered from the air sample col1f^on) the
5/15/79. On this date, the sampler was located about 70-73 meters Tr" the
center of the nearest operating surface aerator. On all other runVating
sampler was located 20-25 meters from the center of the nearest °PVe air
surface aerator. Since at least one type of phage was recovered from
samples collected in ELH on each of these runs, the greater
aerator to sampler was probably responsible for the absence of
the 5/115/79 upwind sample.
108
-------
TABLE
ample
Hafp
/15/79
/19/79
/17/79
J/21/79
10/2/79
46. VIRUS RECOVERY
/U'r samp/es
(PFU/m3 air sampled)5
Sampler^
position
Upwind6
Downwind
Upwind
Downwind
Downwind
Upwind*1
FROM MUSKEGON AIR*
Filtered Filter
supernatant eluate
0*
0
0
0
0
0
0
0
0
0
0
o
AND HATER SAMPLES
•• Water Samples1-
(PFU/liter)
Raw influent
Filtered Sediment
supernatant eluate
50
60
360
267
80
0
o
40
0
27
Filtered
supernatant
0
40
0
27
14
PIMMD station
Filtered
supernatant
0
0
0
0
9
aThe XM2 Biological Sampler/Collector processes 126 cubic meters of air in a 2-hour sampling period
bPFU, Plaque-forming units.
cSamples were assayed by the Centrifugation-Filtration Method.
dThe sampler was located 20-25 m from'the center of the nearest operating surface aerator on all
sample dates, except as noted.
eThe sampler was located 70-73 m from the center of the nearest operating surface aerator.
fO indicates no viruses detected.
9No sample collected.
Sampling run conducted at night (11:00 P.M. to 3:00 A.M., 10/3/79).
-------
TABLE 47. VIRUS RECOVERY FROM MUSKEGON RAW INFLUENT SAMPLES8
Sanple
Date
5/15/79
6/19/79
7/17/79
8/21/79
10/2/79
Centrifugation-Filtration Method15
(PFU/liter)d
Filtered
Supernatant
50
60
360
267
BO
Filter Sediment
Eluate Eluate
0
0
0
0
0
0
0
40
0
27
Total
50
60
400
267
107
Menbrane
Processed
Clarified
Filtrate
Only
Not Done
14
198
26
127
Clarifying
Filter Eluate
1 2
0
28
310
14
Filter fldsorption-Elution
(PFU/liter)
Processed Neutralized
Clarified ftcid
Filtrate Filtrate
0 0
0 13
0
0
Methodc
Concentrating
Filter
Eluate
0
0
=====
Total
0
41
— i— CF1IK PFTPRIOR/VTED
42 78
0 7
10
0
16
0
456
21e
Samples were stored at -70°C until assay could be done. The centrifugation-filtration method was perfonrBd within one week
of the return to Cincinnati. The adsorption-elution method was done at a later date.
F PfT'e?t ^Bef fxtract (0xDla Lst} Lanco Powder) in Mcllvaine's citric acid-dibasic sodium phosphate buffer, pH 7.0, was used
ror aii elutions in this method.
CThree percent beef extract (Oxoid Lab Lanco powder) in Mcllvaine's citric acid-dibasic sodium phosphate buffer, pH 7.0, was
used for all elutions in this method, except as noted. . «~ *• **
. "•
TTU, Plaque-forming units.
0 indicates no viruses recovered.
** Mcllva^e>s citrlc acid-dibasic sodim pl^sphate buffer, pi, 8.0, was
-------
TABALE 48. COMPARISON OF THE COLIPHAGE RECOVERY FROM MUSKEGON AIR SAMPLES3 USING THE SOFT AGAR
OVERLAY METHOD AND KOTT'S MPN METHOD
Sample
date
5/15/79
6/19/79
7/17/79
8/21/79
10/2/79
Sampler
position*'
Upwinde
Downwind
Downwind
Upwind
Downwind
Downwind
Upwind^
Collecting
liquid
ELHf
DWn
ELH
ELH
ELH
ELH
ELH
I , — — *_—
E.
strain C
Over 1 ay
method0
09
0
2.08
0
1.59
0
0.79
CoT/phage recovered using phage host:
coli 13706
, ox-174 host)
MPN
method^ *
0
0
9.0
0.42
2.22
2.04
1.17
E. col
(C-3000,
Overlay
method*
0
0
0
0
1.59
0.86
0
i 15597
n host)
MPN
methodd
0
0
0.033
0
0.13
0.85
0.19
E. coli
(strain B,
Overlay
method0
0
0
0
0
0
0
0
11303
Tl-7 host)
MPN
method^
0
0
0
0
0.11
0.21
0.31
aihe XM2 Biological Sampler/Collector processes 126 cubic meters of air in a 2-hour sampling period.
bjhe sampler was located 20-25 m from the center of the nearest operating surface aerator on all
sampling dates, except as noted.
cValues are expressed as plaque-forming units per cubic meter of air samples.
^Values are expressed as Most Probable Number per cubic meter of air sampled.
eThe sampler was located 70-73 m from the center of the nearest operating surface aerator.
^Sterile Earle's Lactalbumin Hydrolysate without antibiotics.
90 indicates no coliphage detected.
"Sterile distilled water.
^Sampling run conducted at night (11:00 P.M. to 3:00 A.M., 10/3/79).
-------
More E. coli 13706 coliphage were recovered than either of the other-
two types." ThlTTargest amount of coliphage (9 MPN/m3 air sampled, June i»»
1979 sample) was recovered in ELH, using the MPN test and this host.
this air sampling run, the sampler was positioned downwind approximately w
of the time (Table 49), in a fine mist that could be felt. Using this host,
all three downwind MPN levels from samples collected in ELH were higher tna
the three upwind samples. The results were more erratic with the overlay
method, with phage recoveries from 2 downwind samples and V upwind sample-
However, the upwind sample was taken at night (October 2-3, 1979) when en-
vironmental stresses (high temperature, solar radiation, increased wind
velocity, dessication, etc.) would be less than encountered during the day.
Using E. coli 15597, coliphage were recovered in the MPN test from an
three downwind samples collected in ELH. The only upwind sample in whicn
they were detected was the sample collected at night, when environmental
conditions would be more favorable for survival than during the day.. Con-
phage were detected only in downwind samples with the overlay metnoa.
Using E_. coli 11303, coliphage were recovered only using the MPN tes •
The upwind n igfvt~s ample (October 2-3, 1979) had a slightly higher level tna
the downwind sample collected earlier that same day. Because these conPn »
were present in lower amounts in the wastewater than the other two types*
lower amounts would be expected in the aerosols. Although the E_.
11303 levels were lower than the E_. coli 13706 levels, they were v
to those of IE. coli 15597 coliphage. In only one case was the E.. coli
coliphage level considerably higher. This may indicate that E_. coTTlag
coliphage have a greater die-off rate due to environmental stresses and/or
collection and freezing than E_. coli 11303 coliphage.
Kott's MPN method was found to be more sensitive and consistent in
tecting low levels of coliphage in the air samples than the soft agar
method. No E_. coli 11303 coliphage were recovered using the overlay me
but in 3 out 7 cases, they could be recovered with the MPN method. This
probably due to the growth of the coliphage by overnight incubation witn
host prior to assay on freshly seeded plates.
All three types of coliphage were found in all raw influent hie
samples, but levels decreased in the lagoon and pump station samples Uf^ n
50). In general, the quantity of each type of coliphage present in MusKeg
foam (Table 51) and wastewater samples can be expressed in the following
relationship:
NUMBER OF
E. COLI
"13706
j;OLIPHAGL
NUMBER OF
E. COLI
15597"
.COLIPHAGE
NUMBER OF
E. COLI
~1130T
COLIPHAGE.
amount
Concentrations of coliphage in foam samples varied, depending on the am°" ^
of foam collected and the type and amount of diluent, but the undiluted t .
sample, assayed using E. coli 13706, 15597, and 11303, contained 2.85 x i« '
1.8 x 103, and 4.0 x 102 MTper ml coliphage, respectively. Although i»
112
-------
TABLE 49. SUMMARY OF WEATHER CONDITIONS AT MUSKEGON DURING AIR SAMPLING RUNS
'Sample oale
(and lime)
V 15/79
(9:42 AM to
1:50 PM)
6/19/79
(2:00 to
8:10 PM)
7/17/79
(3:45 to
7:50 PM)
8/21/79
( 10:47 AM
2:53 PM)
10/2-3/79
(4:11 to
8:00 PM)
(II:UO PM
3:00 AM)
Sampler
pus i i ton
Upwind
about 9UX
of the
time
Downwind
doout OCX
of the
time, in
the mist
Upwind
about 95X
of the
time
Downwind
about 90%
10 of the
time, in
the mist
Downwind
almost
all of
the time
to Upwind
most of
the time
Prevail iwj
Wind direction
From the NW
From the SE
From the NW
From the NE
to SE
From the E
to NE
From the N
to NW
RJIHJP of
wind velocity
IU-2U mph with
stroinj gusts
to 3D mph
10-20 mph
with many
yusls
0-10 mph, in-
creasing to
10-20 mph
later
0-13 mph,
occasional
yusts
0-10 mph,
occasional
gusts
Almost
none
: Average d.ii ly1
R.iwji.' of barometric
Temperature rot.il.ive pressure -
range (°F) Inimidity (X) (inches of lly)
bO-07 3'J-56 29,-W
01-85 4b-!>2 29.43
76-01 38-52 29. 511
79-83 5U-G3 29.40
69-72 63-74 29.04
About 'jO 74-79 29.04
•-
Additional weather
observations
Partly cluudy early with
w.jrm, sunny and bright.
conditions Liter; nearby
irrigation rigs may h.ive
had some effect on sample.
Sunny, hut, wind/, bright
with slight haze; irrigation
rigs had no effect on sample.
Sunny, bright, hot, some
clouds; irrigation rigs had
a questionable effect on
sample.
Sunny, bright, hot, slight
haze, few clouds; irrigation
rigs had little effect on
sample.
Heavy rain in very early
morning, cloudy, overcast,
and warm later; irrigation
rigs may have had some
effect on sample.
Cloudy, cold, some fog.
aiaken from the monthly data sheets of the National Weather Service office at the Muskeyon County Airport.
-------
TABLE 50. COLIPHAGE RECOVERY FROM MUSKEGON WATER SAMPLES USING THE SOFT AGAR OVERLAY METHOD
*
Sarrple
Site
Cation
rs.sin (Raw
en.
V.'OTt
L.ioon
riryp Station
• Nitrogen
' Added)
i Purrp Station
! (After
j Nitrogen
. Added)
Phage Host
Used
(E. coli
A1OC No.)
13706
15597
11303
137C'
15597
11303
13706
15597
11303
13706
15597
11303
Coliphage Recovered {PFU/liter)a Fran
4/5/79
1.45xl05
1.22xl05
b
4. 77x1 O4
6.45xl04
b
d
d
d
d
d
. d
5/15/79
2.05xl05
l.llxlO5
4.50xl03
S.OOxlO2
2.00xl03
0
d
d
d
d
d
d
6/19/79
S.OSxlO5
8.80xl04
3.25xl03
l.OOxlO3
0
0
S.OOxlO2
0
0
l.OOxlO3
0
0
Taken On:
7/17/79
3.68xl05
8.85xl04
l.OSxlO4
oc
0
l.OOxlO3
0
0
0
l.OOxlO3
S.OOxlO2
0
Sanple
8/21/79
1.84xl05
4.90xl04
e.eoxio3
3.00X103
3.30xl02
0
d
d
d
S.OOxlO3
0
0
10/2/79
5.53xl05
1.32xl05
8.03xl04
3.00xl03
0
0
d
d
d
d
d
d
f Plaque-forming units.
done.
C0 indicates no colip^age detected.
t~No sanple collected.
-------
TABEL 51. COLIPHAGE RECOVERY FROM MUSKEGON FOAM SAMPLES USING THE SOFT AGAR OVERLAY METHOD
.,— —
Sample
Date
• • •
5/15/79
6/19/79
7/17/79
8/21/79
10/2/79
Type
Suspending
Liquid
ELHC
DW
-------
eluate of the foam sediment from the road adjacent to the aeration basin
contained no coliphage, the eluate of the foam sediment inside the aeration
basin contained 3.7 x 104 PFU £. coli 15597 coliphage per gram of sediment.
Bacteria, ranging from 86-7143 colony-forming units (CPU) per cubic
meter of air sampled, were recovered from the air samples (Table 52). The
largest amount (August 21, 1979 sample) was recovered during a run when the
sampler was positioned downwind approximately 90% of the time in a mist that
could be felt. The next largest (5254 CFU/m^ air sampled, May 15, 1979
sample) was obtained during a run when the sampler was upwind most of the
time. However, the three irrigation rigs north of the aeration basin (and,
therefore, upwind of the sampler) were operating during this run, and there
were frequent large gusts of wind, up to 30 mph or more. In addition, both
samples were collected, for the most part, in the morning when aerosolized
microorganisms would have less exposure to heat, desiccation, and solar
radiation than at later times in the day. The upwind night run, collected
on October 2-3, 1979, was slightly higher than the downwind run done earlier
that same day. This is probably due to the much cooler temperatures and the
absence of solar radiation. Since all of the plate counts were done on
frozen air samples, some bacteria were probably lost The lowest amounts
recovered (upwind and downwind, October 2-3, 1979) were probably due, in
part, to the cleansing action of a heavy early morning rain on the air.
_ ELH appeared to be a better collection liquid than distilled water.
The sample collected in ELH on June 19, 1979 (Table 52) was found to contain
nearly 70% more bacteria than that collected in distilled water that same
day. No coliphage (Table 48) were recovered from the distilled water sample.
? nil Lu types of coliphage were recovered from the sample collected
in ELH. ELH appeared to help bacteria and coliphage survive the stresses of
sample collection and freezing. survive
As in the case with virus and coliphage levels, total bacterial counts
decreased as the wastewater progressed through the treatment facility to the
irrigation rigs (Table 53). In 4 out of 5 cases, standard plate counts on
diluted foam samples were only one log lower than those of the corresponding
se o e
(Table 54)' One additional diluted foam sample was
» 7nS,ai?e amount of bacteria> and an undiluted sample con-
?nf? *X I0 V™^1' which 1s 3.5 logs higher than its corresponding
f H n oLSf Ple' Considei"in9 the fact that the diluted samples con-
tained 10-20% foam or less, it appears that the aeration basin foam may
either concentrate bacteria from the wastewater or may exert a protective
effect on bacteria in the frozen samples.
are l^d'in^hi "« di:?ajvanta9es of the XM2 Biological Sampler/Collector
r£Ml5 h JE Table 55. A though particles ^2/um in diameter were not
«UP LrJ laK-er' S^P^e were collected from the aerosols at this
site. Because of this and because some coliphage are similar in size to
some enteroviruses (e.g., polio is similar to fl coliphage (87). it aPPear$
that the sampler would collect animal viruses if the aerosol levels were
large enough and/or if the sampling periods were long enough.
116
-------
JABLE 52. RECOVERY OF BACTERIA FROM MUSKEGON AIR SAMPLES9
Samp 1 e
date
5/15/79
6/19/79
7/17/79
8/21/79
10/2-3/79
3TL. . ~ •
Sampler^
position
Upwind^
Downwind
Downwind9
Upwind
Downwind
Downwind
_ Upwind"
Standard plate count0
(CFU/ml s ample )d
3310
127
64
124
4500
33
42
CFU/m3air
sampled6
5254 (149)
212 (6.0)
126 (3.6)
305 (8.6)
7143 (202)
86 (2.4)
99 (2.8)
he XM2 Biological Sampler/Collector processes 126 cubic meters of air in a
'-nour sampling period.
y>e sampler was located 20-25 m from the center of the nearest operating
CR ce aerato>" on all sample dates, except as noted.
sacteriai assays were done on frozen samples which had been collected in
„ ?-n1e Earle's Lactalbumin Hydrolysate without antibiotics, except as
noted. . . . •
Q^ Ci i • r
erp ^^"y-forming units.
fTh Sir samples snown in parenthesis. :
he_sampler was located 70-73 m from the center of the nearest operating
aerator.
Sample was collected in sterile distilled water.
Jj^ sampling run was conducted at night (11:00 P.M., 10/2/79, to 3:00 A.M.,
117
-------
00
TABLE 53. STANmRO PIATE COUNTS OF MUSKBOON WATCH SAMPLES9
Sanple
Date
4/5/79
5/15/79
6/19/79
7/17/79
8/21/79
10/2/79
Bacterial Counts (CFU/ml)b At These Sampling Sites:
Aeration Basin
Raw Influent
4.65 x 105
4.73 x 105
1.76 x 106
3.19 x 105
2.75 x 107d
3.35 x 105
8.90 x 106df
1.67 x 104
West Lagoon
1.10 x 106
3.25 x 104
1.94 x 105
4.10 x.102
3.50 x 10M
e
6.73 x lO3^
2.15 x 102
Pump Station
Before
Nitrogen
Addition
c
c
6.00 x 103 1
3.75 x 10? 5
c 4
c i
c
c
After
I'i trogen
Addition
c
c
.83 x 104
.50 x 102
.60 x 10*1
.11 x 103
c
c
Assay done in Cincinnati, Ohio, on frozen sanples, except as noted.
CFU, Colony-forming units.
So sample collected.
Assay done in Muskegon, Michigan, 03 scon ag possible after sample collection.
wot done.
Plates counted at 90 hours.
-------
TABLE 54. COMPARISON OF STANDARD PLATE COUNTS3 ON MUSKEGON FOAM
AND AERATION BASIN RAW INFLUENT SAMPLES
Sample
date
5/15/79
6/19/79
7/V7/79
8/2-1/79'
10/2/79
Suspending Final volim
liquid of foam san
of foam (ml)
ELHd
DWe
ELH.
10* BE, pH 7.0f
ELH
None (undiluted)
ELH
50
50
100
150
1 50
9
150
neb
iple Bacterial counts (CFU/ml)c
Foam
1.19 x 10*
3.9 x 103
2.55 x 104
3.25 x 105
4,8 x 104
7.65 x TO7
3.46 x 103
Raw
4.73
1 .76
3.19
3.35
1.67
Influent
x iq5
x 106
x TO5
x 105
x 104
aAssay was dotie on frozen raw influent and foam samples "in Cincinnati, Ohio.
''Foam samples were collected :in a -s'terl le beaker ,: Sl:1oweil:tb settle, and
were made up to a final volume with the chosen suspending liquid.
CCFU, Colony-forming units.
^Sterile Earle's lactalbumin hydrolysate without antibiotics.
eSteri'le distilled water.
beef extract in Mcllvaine's citric acid-dibasic sodium phosphate
buffer, pH 7.0.
applicable.
119
-------
TABLE 55. THE ADVANTAGES AND,DISADVANTAGES OF THE XM2
BIOLOGICAL SAMPLER/COLLECTOR
Advantages
1. The XM2 Biological Sampler/Collector processes a large volume of
air (1050 liters/minute).
2. The air unit can be removed from the sampler for ethylene oxide
sterilization.
3. Many of the particles in the respirable range can be collected.
4. Particles are collected in liquid, thereby minimizing the
dehydration of airborne viruses and bacteria and increasing their
chances of survival.
5. Particles are collected by a combination of scrubbing and
impingement in the wet collector of the sampler.
6. No arcing or other electrical problems occur with this sampler.
Disadvantages
1. Sterilization procedures cannot be done quickly.
2. The air unit cannot be autoclaved.
3. Sterilization cannot be done in the field.
4. The sampler is complicated, and the adjustment of the peristaltic
pump is difficult. Considerable time is necessary to become
familiar with the sampler.
5. The sampler does not collect the particles <2 /im or >12yum in
diameter.
6. Two people are needed to lift the cumbersome and heavy sampler
in and out of the transporting behicle. However, it can be moveo
quite easily once it is situated on a wheeled platform at the
sampling site.
7. An adequate supply of electricity is needed at the
120
-------
Possible explanations for the failure to recover animals viruses are
following:
!• Raw influent virus levels may be too low to produce the aerosol
levels necessary to be collected with this sampler.
2- Viruses may not be aerosolized due to adsorption to wastewater
particulates and/or entrapment by the blanket of foam on thesurface
of the aeration basin.
3- Viruses may not survive aerosolization by the aerators, dessication
and debilitation by environmental factors, and damage during the
passage through the sampler air unit and tubing to the collection
liquid.
4- Viruses may be present in the particles not retained by the XM2
Sampler; i.e., in those <2>um and >12/unfTn" diameter. In order to
collect these, the sampler design would have to be altered, thereby
affecting the air flow through the air unit and the separation of
the 2-12 ,um particles. Further, as currently designed the air
passages for these particles (<2/jm and 7-12>um) cannot be
sterilized.
5- A longer sampling period (perhaps several days to several weeks)
may be necessary to prove the presence or absence of animal
viruses. An alternative to this would be to pool air samples from
multiple samplers and/or sampling runs.
6- Cell systems, other than' BGM, may .give better recovery of viruses
at this site.
7- Loss of virus may have occurred during storage and assay pro-
cedures. For example, viruses may adhere to sample containers and
Pipettes, become entrapped on membrane filters, in spite of pre-
treatment, or may die during freezing and thawing of the samples.
121
-------
SECTION 8
BACTERIAL AEROSOL ENUMERATION AND IDENTIFICATION AT A
WASTEWATER SPRAY IRRIGATION SYSTEM
The purpose of this project was to collect, enumerate and identify
bacteria in aerosols at the Muskegon County, Michigan Wastewater Manageme
System Number 1. The description of this facility is presented in Sectio
/ •
MATERIALS AND METHODS
Air Sampling
Aerosols are particulate materials in either solid or liquid form
and may also include gases and vapors that are adsorbed or contained in
airborne particles or liquid droplets. Inhalation is a possible route OT
infection because viruses and most pathogenic bacteria are in the res-
pirable size range. The health hazard posed by aerosolized particles
depends in part on their ability to deposit in the lungs. The most im~ i
portant factor in lung deposition is the size of the particle. The part1
size is usually expressed as mass median or aerodynamic diameter. The
aerodynamic diameter is a function of both the physical diameter and the
density of the particle. It is defined as the diameter of a unit dernsw
sphere having the same settling velocity as the particle in question ot
whatever shape and density. Usually particles with an aerodynamic
greater than 30jum do not enter the nasal passage, those with an a
diameter ranging from 5-30 jum are deposited in the naso-pharyngeal regio •
Particles ranging in aerodynamic diameter from 1-5 Aim are usually deposi
in the tracheobronchial region by sedimentation. Particles less than '
urn in aerodynamic diameter are deposited in the pulmonary or alveolar ti
region by diffusion. It is the last category of particles that may cons
a health hazard by inhalation. Particles deposited in the tracheobroncm
region can be removed by muco-ciliary action (spiral movement of the
mucus by ciliated epithelium) toward the trachea and pharynx where the
material is swallowed or expectorated. The swallowed particles then may
pose a health hazard via ingestion by exposing the gastrointestinal trac
to the pathogens.
The Andersen six-stage cascade impactor (87,88) was used in this
project (Figure 13). The Andersen air sampler collects and separates
particles into different size ranges and thus provides a good size dle
bution of the particles. The Andersen sampler is used only when partic' ters/
concentrations are high, since it has a limited sampling range of 28.3
122
-------
STAGE NO.
\\\\ /777
0.0465" DIA.
3.54 FT/SEC
STAGE 2
0.0360" CIA.
5 S9 FT/SEC
STAGE 3
CC280" DIA.
9 74 FT/S£:
STAGE 4
0 G2IO" DIA.
17.31 FT/SEC
STAGE 5
C.OI35" DIA.
41.92 FT/SEC
STAGE 6
C.OIOO" DIA.
76-40 FT/SEC
Figure 13. Schematic diagram of a six-stage Andersen sampler.
123
-------
min. The perforations become smaller with each succeeding descending
stage, so that as a pump pulls one cubic foot of air per minute through
the sampler the particles will be sized aerodynamically such that the
respirable (stages 3-6) versus nonrespirable (stages 1-2) particles can
be separated.
The samplers were loaded using aseptic technique and autoclaved be-
tween use to minimize carry-over contamination. The closed samplers were
then moved to the sampling sites.
Sampling Sites
The sampling sites were chosen to evaluate the contribution of various
treatment units and to determine aerosol levels in worker areas. Upwind_
and downwind samples were taken in the irrigated fields and at the aeration
basins throughout the growing season. Air samples were also taken at the
pump stations, the farm, the administration building and also at the
nearby County Road Commission just outside the Wastewater Management
System (Figure 14). The samplers were run at one cubic foot per minute
for ten, five, or two minutes depending on the number of colony forming
units (CPU) found in previous runs.
Enumeration and Gram-Negative Bacteria Isolation and Identification
The primary cultivation medium used in the Andersen sampler was
Trypticase Soy Agar (TSA), a nutritional medium. The exposed plates we£e,
incubated for 24 hours at 35°C. The colonies on these plates were counted,
and plates three through six were replicated using velvet onto two more
TSA plates-which were again incubated for 24 hours at 35°C. These submaste
plates were then each replicated onto three different media: sheep blood
and MacConkey's agars for identification; and KF Streptococcal, M-endo,
M-FC, and XLD agars for enumeration of fecal indicator organisms (30).
The colonies which grew on MacConkey's agar were considered to be gram-
negative colonies, and the API-20E system of 21 biochemical tests were useo
to identify the isolates. The colonies which did not grow on MacConkey s,
but did grow on sheep blood agar, were considered to be gram-positive,
tentatively, and worked up in another fashion. A flow diagram for this
enumeration, isolation and identification is presented in Figure 15.
Gram-Positive Bacteria Identification
There was an initial problem with the identification of the gram-posit1
bacteria, since the soil organisms present in the air were interfering
with the tests. This interference was avoided by using selective media
to eliminate the soil bacteria. Thus, KF-Streptococcal Agar was used to
select out for streptococci, and Mannitol Salt Agar was used to select
out for staphylococci (30).
B and 0 disks were used to identify group A beta streptococcus an(?..tion
Streptococcus pneumoniae, respectively, after they showed rings of innib,JL
on Sheep Blood Agar at 35°C for 24 hr. Any negative cultures were streaKeu
onto Bile Esculine Agar and incubated for 35°C for 3 days, checking daily-
124
-------
I KILE
Adm.
T.P.
SC
R.C.
GR
FR
I
II
Administration Building fit Stumo
Trailer Park
School
Road Commission
Grainery
Farm Equipment Storage
Pump I
Pump II
Figure 14. Map of Muskegon Wastewater Management System
no. 1 sbbwing the air sampling sites.
125
-------
ro
en
TRirr.JCASE SOY AOAR
I
REPMCATK OH
I
TRIPTICASE SOY AGAR
REPLICATE ON
SHEEP BLOOD AGAR
I
TRITTTCAriF. ROY AOAR
——RFn.lCATK ON——
KF STREPTOCOCCAL AGAR
(Fecal Streptococci)
m-ENDO AGAR
(Total Coliforms)
xr.D AGAR
(Shiegella)
MAC CONKEY AGAR
m-FC AGAR
(Fecal Coliforms)
IDENTIFY USING
API SYSTEM
(Gram-Negative)
FURTHER
IDENTIFICATION
(Gram-positive, see
Figure 16)
Figure 15. Flow diagram for enumeration and gram-negative bacteria isolation and
identification scheme.
-------
Inf., • e Escul™e positive cultures were inoculated into Brain Heart
innr !°n Broth and incubated at 35°C for 24 hr> The 24 hr. culture was
for S d 1nto Bra1n Heart Infusion-Agar.with 6.5% salt and incubated
tn K • 7 days at 35°c> If there was 9rowth here, the culture was considered
tu oe either Streptococus faecal is or faecium.
. Any culture which grew on the Mannitol Salt Agar was inoculated into
<>rain Heart Infusion Browth at 35°C for 24 hr. A drop of this culture
1.1 K *•••**•* iv*( V I W ** WI I U w w */ w I VI fc» ~ III* n VHWV VI v II I •? WUIwUfC
in o^nd to 0-49 m^ °f rabbit coagulase plasma, covered, and incubated
a 3/oc water bath for 24 hr. If the tube was clotted, the culture was
led Staphylococcus aureus.
A flow diagram of the replication and identification scheme for
•positive bacteria is shown in Figure 16.
RESULTS
at 'ne proportions of the respirable CPU recovered on the various media
.the treatment plant sites sampled and the mean respirable CPU at these
sites are shown in Table 56.
n The downwind-air from the aeration basin area contained the highest
moers of colonies on M-Endo, M-FC, and MacConkey agars, and Colony
rmng Units (CPU) in the respirable size range. Although the samples in
e fields were taken upwind and downwind, there appeared to be a definite
factS"°Ver of air from anotner or the same rig. This can be seen in the
as H- tlie uPw^nd an^ downwind rig sample values were closer and not
Different as the values found upwind and downwind of the aeration
bepn"15' Ttie background values taken in the fields after the rigs had
nums °Ut °^ °Peration for several weeks were.much lower. The proportional
i °f ttie various respirable bacterial organisms identified in the
'es grouped according to location are presented in Figures 17 to 21.
Presented in these figures represent different number of samples
ed and because each sample collected represented a particular
of air, the total volume of air, M3, from which the identifications
made is included in each figure.
t There was found to be an increase in the number of bacteria found in
numKSUrroundin9 a1r as a result of the wastewater site. The increased
roDers of bacteria found downwind of the aeration basin and the irrigation
conf 1n the fields, suggested that these areas were sources of bacterial
of ruination of the air. The population of aerosolized bacteria downwind
aeration basin and the irrigated field rigs did have a higher
age of gram-negative bacilli than the upwind samples, suggesting
the areas were also a source of gram-negative bacilli.
hi The population of aerosolized bacteria in the downwind air had a
In A percentage of fecal-indicating bacteria than the control samples.
"addition, the population of aerosolized bacteria in downwind samples
bartnave a higher percentage of pathogenic bacteria (defined as gram-negative
°cteria and gram-positive cocci) except for Staphylococcus aureus and
127
-------
GRAM POSITIVE
00
TRYPTICASE SOY AGAR
1
KF STREPTOCOCCAL AGAR MANNITOL
B DISK
POSITIVE
GROUP A
BETA
STREPTOCOCCUS
BILE
1
O DISK BRAIN HEART 1
1
POSITIVE
STREPTOCOCCUS COAG
PNEUMONIAE
NEGATIVE 1
1 NEGATIVE
ESCULIN AGAR (UNIDENTIFIED)
BHI
BHI & SALT
SALT AGAR
^FUSION BROTH
LJLASE
1
POSITIVE
STAPHYLOCOCCUS
AUREUS
NEGATIVE POSITIVE
(UNIDENTIFIED) STREPTOCOCCUS FAECALIS
STREPTOCOCCUS FAECIUM
Figure 16. Flow diagram of the replication and identification scheme for
gram-positive bacteria.
-------
TABLE 56. ENUMERATION OF, RESPIRABLE COLONY FORMING-UNITS
RECOVERED AT VARIOUS SITES
~~~~weans or total colony forming units ot
respirable size that were followed per
cubic meter of air
Media/ type of bacteria
M-Endo/total M-FC/fecal -'McConkey/ TSA/
Source* coliforms coliforms -qr am- negative total
Aeration basin (1) upwind** ""
( 1) downwind**
Field. rigs ( 18.3) upwind
(18. 3)1 down wind
.U&.3.) background
East & west lagoon (1)
Farm storage (200)
Pump station I (6)
Pump station II (6)
Administration Building (400)
Road Commission (400)
Trailer park (200)'
Grainery (400)
Inn (17500)
School (440)
51
1100
21
35
0
0
0
0
0
0
o
;is
0
0
100
15
1600
28
35
0
100
0
0
18
35
0
18
70
0
0
140
2200
110
290
70
100
18
35
100
70
11.
260;
0
70
140
490
2800
660
700
320
380
230
620
350
500
370
380
320
240
350
*Approximate distance (meters) from edge of nearest source (aeration basin
or spray boom - see Figure 14).
**0ne sample collected at 18.3 meters was in the same range as the others.
129
-------
Staphvtococci
aureus 8
Pasteurella spp. 2
Serratia
spp. 4
Salmonella'
spp. 2
Citrobacter
spp.12
Pseudomonas spp 21
Efltarobacter
Enterobacter cloacae 38
Klebsiella spD. 25
Other
Pnterobacter
spp.10
Figure 17.
Relative proportions of respirable bacteria
identified downwind of aeration basin in 2.06
cubic meters of air.
130
-------
Cltrobacter
spp. 1
Streptococcus
faecium or
faecatis 1
Staphvlococgng
aureus 1
Pseudomonas spp. 4
Enterobacter cloacae 4
Group A Beta
Streptococcus
Klebsiella spp. 2
Figure 18. Relative proportions of respirable
bacteria identified upwind of aeration
basin in 2.01 cubic meters of air.
131
-------
gqrratta spp. 1
Aeromonas
hydrophlla
Streptococcus
.faecium or
faecalis 4
Staphvlococcus
a u re us 2
Enterobacter aaalomerans 7
r cloacae 2
'-^
Other Enterobacter spp.
Pseudomonas spp. 13
Group A
Beta Strep 2
Klebsiella SPP. 3
Pasteurelia
spp. 2
Figure 19. Relative proportions of respirable bacteria
identified downwind of field rigs in 3.97
cubic meters of air.
132
-------
Group A Beta Streptococcus 3
Enlerobacter cloacae 1
Streptococcus faecalls or
^•™»-"^^» MHH^MM
faecium 7
Figure 20. Relative proportions of respirable bacteria
identified upwind of field rigs in 3.97
cubic meters of air.
133
-------
Aeromonas
hydrophila
" Enterobacter
aqqlornerans
4
Streptococcus faecalis
or iaecium 8 | / / Enterobacter
cloacae. 3
Staohvlococcus aureus 2
Group A Beta Streptococcus 7
Pseudomonas spp. 18
Figure 21. Relative proportions of respirable bacteria identifie
at other sourcesa in 7.46 cubic meters of air.
a. East and West lagoon, farm storage, pump I, pump II, administration
stump, road commission, trailer park, grainery, inn, and school.
134
-------
Streptococcus faecal is or faecium. .Pseudomonas spp. was relatively more
aBunaiffl! at sampling sites"awayTrom the aeration basin and tne neiub
(Figure 21).
DISCUSSION
. Focussing on the results from others, Fair ^.^"f^Sn^anks^nd
total bacteria count of 787 CFU/m3 at activated sludge aeration tan
tackling filters using the Wells Air Centrifuge Usjng Andersen
sampler at a trickling filter, Albrecht (91) found 50bU uu/m a
JFU/m3 of total bacteria at 15.2 meters down^"J-?7^fl?Fm at an activated
Andersen samplers found total bacteria counts of ^ ^ 5150 CFU/m3
sludge unit/1000 to 3040 CFU/m3 at a pickling filter and bio
at a nrax^ln^ ,,«^ rarnnw et al. (93) found J/D tru/m ui
a preaeration unit. Carnow et al. (93) l°u™ *' ° ^ ' Andersen samplers
, °
r; t •
Cr? ifornia, Johnson et al. (95) Using high v ol«« W le^Q CFU/m3 Of
CFU/m3 of total bacteria, 43 CFU/m3 Of total col forms, o. ^^^
fecal coliforms, 2.3 CFU/m3 Of feca streptococci 240 uu^m^
80 CFU/m3 of ^cobacterium, 4.3 CPU n,3 of. Clos^ndium S^-^^^.^
C?^/m3 of Streptococcus aerus and .4.1 CFU/mj JjgBSjlL^ ^ (96)
20 m from the wet-line etfgTof spray ^rigators. eaus lla us1ng
.Jjund .10.000 CFU/m3 of total .bacteria and 50 CFU/m °' 17^^with
Andersen samplers 46 m downwind at a golf course spr a/
wastewater.
- -'^
Jn activated sludge site and found that 6X of "" sampl^rTToUnT
^2542 CFU/m3).9 Cronholm (98) «i"9 2-stage Anderson P^ ^^
^at 18% of the enteric isolates were K. fiS|gg"g * 3.
The maximum total enteric counts averaged about 8UU per
.. The traditional coliform indicators for water P°JJuJ0JnSurvive as
Jicators of bacterial air contamination Because tney Johnson et al.
fi" in air as, for example, the encapsulated ™3£ZL- 1table 1n-
(89) recommended that fecal streptococci would-be a J0. It can
f cator, while Randall and Ledbetter (97) ^commena
be lt human bre athing
caor, while Randall and Lee Tnffoar per
b,e calculated that an average adult human bre athing rf Qf th
b/eath and at a rate of 20 breaths per minute, stana 3 ^ gram_ gative
n basin would inhale about 28 total ^spira grOT.negative
ble bacteria per minute. s^enteen perc ew dose for man of
would be Klebsiella spp. The minimum inrec
spp. is not known.
„ , The most significant finding of ^ibciella^pp."'?^ the atmosphere
Relatively high levels of respirable ^J^f^fr^Pother recent studies
downwind of wastewater sources compareo^to severa.
-------
(93-95). Levels found in this study were comparable to those of Cronholm
(98) but lower than those of Ledbetter (97).
Klebsiella pneumoniae itself can be found in wet areas around the
home such as kitchen sinks and drains (99). It has been found in wooden
red wood reservoirs containing finished drinking water and from freshly
cut logs (100). Sixty to eighty (60-80) percent of the total col i forms
found in raw wastes from pulp mills were identified as Klebsiella ^ ted
The source of Klebsiella at Muskegon is very likely the paper irnTl 'oca
in Muskegon.
It has been found that Klebsiella pneumoniae can survive much longer
than the traditional indicator Escherichia coli and even Proteus wUSifcrr-^
in gas-free mineral water (102). The factors which influenced the mult p
cation and behavior of p. aeroginosa and K. pneumoniae which were corm°"Jjae
isolated from sink tops were studied in FFance (103). Klebsielja pngHi!^T
was more resistant to drying and survived longer than one week! This w
mean that Klebsiella pneumoniae survives longer in the environment incr
the robability f nfecting the human population. It has also been fou.
that bacteria which are found to be resistant 'to certain metals, chemj[p
or antibiotics are also now likely to be resistant to other metals, cne
or antibiotics. The lead-resistant species of Klebsiella isolated from
domestic sewage, for example, was found to have simultaneous resistance
to other inorganic metals and antibiotics (104). Unfortunately contact
with metals, chemicals, and antibiotics and thus resistance to them can
be easily achieved by bacteria at a wastewater treatment plant and the
more antibiotics the bacteria become resistant to the harder they are TU
the clinician to treat.
Health Effects to thelmmunologically Compromised Patient— ,.,„,
It is important to realize that the health effects of any bacter,l"niae
are directly related to the patient's immune status. Klesblelja £n§JfflSU^-
has been found in local infections of patients with both acute leukemia
and granulocytopenia (105). Klebsiella oxytoca was isolated from the -\
blood of a leukemia patient in Japan where it was reported that thein;y
pathogenicity" of the bacterium was observed (106). Gross et al. (J-u/;
performed experiments on animals with complement deplection caused by
cobra venom factor. The experiments suggested that hypocomplementemia
predisposes to bacterial pneumonia caused by K. pneomoniae and other
bacteria. Gross thought that this might explain the unusually high 1
of pulmonary infection which exists in patients who have complement
tothP Snril1' (1??)/?un(? -th* decreasing the amount of available
to the body resulted in impairment of lung clearance of Klebsiella
This was thought to be due to killing by the alveolar macropKSgTor
^e*^ amount of phagocytosis. K1ebsiella has not only been ^ d
in infection of the lungs, but also the ears, skin, horse's genitals an
even gun shot wounds (109). All of these sites a?e sSsceptable to expos"
by crop spraying of wastewater.
136
-------
Ot[ier Health Effects-
the i^ottl?r important point which must be discussed is the possibility of
5e ,£Jj§b_iJ£lla bacterium being an allergen. An allergen doesn't have to
ajip in9» so the bacterial counts might underestimate the amount of
asth 6n' *n studying 19 patients with allergic rhinitis and bronchial
that t 3nd ^king for allergies to Klebsiella pneumonoiae it was found
(llm bacterial allergy was more important than the bacterial infection
susr Jhus tnere is a need for more studies on the immune status of the
SD cePtible human population in general terms, in terms of allergic re-
J]°nse of the bacterium Klebsiella, and in terms of the population's
5Ceptability towards Klebsiella.
137
-------
SECTION 9
SELECTED ORGANIC CHEMICALS IN AEROSOLS AND WASTEWATER AT A
WASTEWATER SPRAY IRRIGATION SYSTEM
Community and occupational health concerns regarding wastewater treat-
ment facilities have generally focused on aerial dispersion of pathogenic
viruses and bacteria and odor discharge from the treatment processes. Re-
cently, accidental or intentional discharges of chemicals from industrial
sources have drawn attention to the presence of toxic chemicals in waste-
water and their potential release into the atmosphere (64,65). The waste
effluents from many industrial installations eventually enter municipal
treatment facilities and eventually are discharged to streams, lakes or
oceans. These effluents can contain volatile contaminants emanating from
raw materials, products, by-products and cleanup operations of manufacturing
processes. Regardless of the source, it is highly probable that some quantity
of volatile or semi-volatile contaminants will enter the waste stream of
many wastewater treatment facilities. The importance of air release of
toxic chemicals from wastewater at treatment plants needs evaluation.
The increasing quantity of wastewater sludges being produced and the
prohibition of ocean dumping of such sludges have led to renewed interest in
wastewater treatment methods that provide reclamation and renovation of
wastewater and/ or sludge by land application. One type of wastewater treat-
ment involving land application of wastewater is currently being practiced
at the Muskegon, Michigan, Wastewater Management System No. 1 (MWMS). A de-
scription of this facility is presented in Section 7 of this report. This
system would be predicted to have a high potential "for release of volatile
chemicals into the atmosphere.
The purpose of this study was to evaluate the release of volatile organic
compounds during the treatment of wastewater at the MWMS, to determine if
significant exposure of chemicals to the workers at the facility occurred
and to determine if significant amounts of chemicals are released to the
atmosphere of the nearby community.
Air release of volatile organics from wastewater treatment facilities
can occur by three mechanisms: evaporation, air stripping and aerosol for-
mation (111,112,113). The treatment processes at the Muskegon system where
air release may occur are aeration, lagoon impoundment and spray irrigation.
All three mechanisms contribute to release at the aeration cells where air
enters the wastewater by means of mechanical surface aerators. Release
the storage lagoons will occur by evaporation. Aerosol formation and
138
-------
evaporation mechanise are responsive for air concentration during the
spray irrigation operation.
EXPERIMENTAL METHODS
Air release of volatile organic Chemicals from wa^ewater^t^eacj
ment process was assessed by c?"curr^^Later samples were used to determine
wastewater at each treatment site. "«^™ available for release into the
the amount of specific chemicals Pg^JJ^S'S each treatment process
air. Collection of influent and effluent sarnies achieved by the
also provided information on the decrease in cone ^ ^ downwind of
particular treatment. Air samP1" w^eenabled determination of the amount
each treatment process. These jampie fr0m the treatment process.
of contaminant in the air that was released
Sampling and Ana ly^ Procedures
mpng an na . effluent of
Wastewater grab samples fr*c°"?^
the aeration basins, at one °ft^% ^Sainage ditch. The wastewater
to irrigation rigs), and from the field ^ainag tewater W1th a gal-
" lss bottle
the aeration basins, a one t ainage ditch. The wasew
to irrigation rigs), and from the field ^ainag tewater W1th a gal-
samples were collected close ^^^"ly transferred into glass bottles
vanized bucket. The samp es were «^^inim1ze the head space. The
aluminum foi,,: and
nmze
• aluminum foi,,
stored under. refrigeration , .
»• «sffasrsi-
rs
Proximately 1 meter above the water surr to tripods located ap
the water surface, or by ""J^^LlJ surface and 2 to 3 ^L^rigs
Proximately 1.5 meters above the water su "ear^the irr gatjon 9
the water. The location of air samples co ^ wd tsSParound Upwind
varied because of the rotation of the r ^ mete ab the grouna ^
lected 5 to 50 meters from the ngs an hg Hgs and 1.5 fflttcr
samples were collected 30 to 50 meter*
the ground. rontaining 100 mg of
samples and
ibrated before and after
139
-------
The sorbents were desorbed by placing the sorbent in 2 raL of petroleum
ether and allowing the mixture to stand for one hour with occasional shaking.
A Bjjl aliquot was then analyzed by gas chromatography with electron capture
detector.
RESULTS AND DISCUSSION
Initial air sampling surveys were conducted during April and May 1979.
The April survey was limited to monitoring near the aeration basins and out-
side the Administration Building located on the site. Spray irrigation was
not being practiced at that time since the fields were being prepared for
planting. The aeration basins and the spray irrigation operations were con-
sidered to have the highest potential for aerial release of chemical con-
taminant from the wastewater. Upwind and downwind samples were obtained at
each treatment process in order to evaluate the extent of aerial release of
substances from the wastewater. Due to fluctuating wind directions, aerial
transport of contaminants may occasionally cause them to reach the upwind
sampling locations.
Table 57 presents the results from the initial air monitoring surveys.
Three chlorinated organics, chloroform, carbon tetrachloride and tetra-
chloroethylene, were found in the samples. The downwind air samples col-
lected at the aeration basins showed significant concentrations of these
substances. In the May survey samples were obtained downwind of spray
irrigation rigs and the storage lagoons. Table 57 shows that only trace
amounts of these substances were present in the air at these locations. Wastewater
samples were not obtained duringthesepreliminary surveys so comparison of
the air concentrations with wastewater levels cannot be made with these
samples.)
Table 58 and 59 show the results of detailed study of the aerial re-
lease of four volatile chlorinated organics from wastewater at the aeration
basins. Influent and effluent wastewater samples as well as upwind and
downwind air samples were obtained during four sampling periods of one to
three days duration each from June through October 1979. The aeration
process markedly reduces the wastewater concentrations of chloroform and
trichloroethane. Trichloroethylene and tetrachloroethylene do not always
appear to be as easily removed. The downwind air concentrations show that
significant quantities of these volatile substances are stripped from the
wastewater during aeration. The influent and effluent wastewater concen-
trations represent the daily means during the air sampling period, and it
should be recognized that the retention time in the aeration basins is several
days. Thus, the difference between influent and effluent concentration may
not necessarily represent the actual amount of material stripped, but it at
least provides an index of estimation.
Air concentration of the chlorinated organics around the spray irrigation
operation are given in Table 60. Relatively low levels of trichloroethane,
trichloroethylene and tetrachloroethylene were found. Chloroform was not
found in either upwind or downwind air samples. In general, the air concen-
trations reflect the relatively low concentration of these substances in the
140
-------
TABLE 57. INITIAL AIR SAMPLING SURVEYS AT MUSKEGON
COUNTY WASTEWATER MANAGEMENT SYSTEM
Concentrat1on,/jg/m3
.Location Chloroform Carbon Tetrachl or ide Tetrachloroethylene
April 1979
Aeration Basin
(Upwind) 17 13 53
Aeration Basin
(Downwind) 189 1098 . 252
Outside Admin. Bldg. 6 32 17
May 1979
Irrigation Rig — 0.3 1.3
Storage Lagoon — NDa 3.1
Outside Admin. Bldg. : -- ND ND
Perimeter of Site — 0.2 0.5
—Not analyzed.
aND - None detected, concentrations are less than 0.1/jg/m3.
141
-------
TABLE 58. DAILY AIR CONCENTRATIONS NEAR AERATION BASINS AND
ASSOCIATED WASTEWATER CONCENTRATIONS
Chloroform
Wastewater Cone., a
,/jg/L
Influent
>/ll/79 892
Effluent
Air Conc.,d
.Aig/m3
Upwind
68 0.5
Downwind
39b
6/12/79
6/13/79
1180
1520
82
75
0.5
0.5
36'
7/16/79
7/17/79
7/18/79.
670
480
800
28
18
22
0.5
0.5
0.5
96 c
202 d
124 d
aThe values are individual samples or means of 2 to 4 samples. •
bChromosorb 102, high flow, basin-3.
cCharcoal, low flow, basin-3.
dCharcoal, low flow, basin-2.
142
-------
.£»
co
TABLE 59. DAILY AIR CONCENTRATIONS AT AERATION BASINS AND INFLUENT AND EFFLUENT WASTEWATER
CONtENlRATlUNS OF THE AERATION BASINS"
Trichloroethane Trichloroethylene Tetrachloroethylene
Hastewater Cone.,
Influ.
7/16/79 56
7/17/79 67
< 7/18/79 60
i
8/20/79 NO
8/21/79 72
10/17/79 4.4
10/17/79 3.5
Efflu.
NDC
12
17
NO
3.0
0.3
0.4
Air Co
>jg/
pwind
0.2
4.6
3.2
7.3
NO
4.7
3C"
ownwind
10
22
23
90
77
17 d
33
astewater Cone.,
jug/L
nflfi:
1.8
69
118
NO
30
7,5
6.7
tfflu.
NO
110
140
ND
12
0.7
1.0
Aiir Conc.,b
/jg/m3
pwind
ND
NO
2.5
0.9
0.8
2.4
3.0
ownwind
11.5
3
73
17
14
8.8
40
Wastewater Cone.,
>*g/L
InfluV
3.0
63.3
92.5
ND
25
7.7
8.9
Efflu.
ND
60.6
88
ND
1.0
0.4
1.0
Air Co
>ug/
pwind
ND
ND
9.4
6.5
0.6
1.8
8.6
nc.,b
ownwind
6.8
34
34
23
8.7
46
American Conference of Governmental Industrial Hygienists (ACGIH) has reconwended a Threshold Limit Value
(TLV) of 1,900 mg/m3 for trichloroethane, 270mg/m3 for trichloroethylene and 670 mg/m3 for tetrachloroethylene.
bAir samples were collected using charcoal and Chromosorb 102 sampling tubes at flow rates of 25 to 60 mL/min.
The values are means of 2-5 samples.
CNO - designated not detected; concentrations are less than O.ljig/ro3; wastewater are less than 0.1/jg/L.
dThis value is the mean of 10 samples with a range of 0.2 to 103/ig/m3.
-------
TABLE 60. OAlir tUNUtfOHflTlUH:> UF SELECTED CHLUKlNATtt) OHGflNlCS NEAR SPRAY IRRIGATION RIGS
irichloroethane
wastewater Cone.,
jug/L
N.P.d
Stat.
7/16/79 Rig 11 1.3
Rig 12 1.3
7/17/79 Rig 11 11
7/18/79 Rig 11 5
11 5
8/20/79 Rig 12 0.2
8/21/79 Rig 12 3
Field"
Efflu.
NDC
NO
5
6
6
0.2
30
Air Cone.,
,/ig/m3
Down
Upwind wind
NO NO
NO 0.2
NO NO
0.8 1.3
0.7 1.3
1.9 2.3
2.8 2.7
Irichloroethylene
Wastewater Cone.,
jug/L
N.P.
Stat.
1
1
122
68
68
ND
11
Field
Efflu.
ND
ND
33
79
79
ND
25
Air cone.,
vug/m3
Down
Ip wind wind
ND ND
0.4 2.6
ND ND
2.2 9.3
0.7 5.0
0.1 0.4
0.5 0.6
ietrachloroethylene
Wastewater Cone.,
Jjg/L
N.P.
Stat.
NO
ND
72
40
40
ND
1
Field
Efflu.
ND
ND
7
43
43
ND
26
«ir tone. ,
/ig/m3
Down-
Upwind wind
ND ND
1.2 2.1
ND ND
1.8 8.6
0.6 4.6
0.3 0.5
0.1 0.1
aSpray rig influent wastewater collected at North Pump Station.
*>Field effluent water collected from drainage ditch that collects leachate from irrigated fields for discharge
to surface waterways. These samples were collected at the same time as the pumping station samples and
therefore represent wastewater sprayed onto the fields prior to the time ot collection ot the pumping station
sample.
CND - designates not detected; air concentrations are less than 0.1/jg/m3; wastewater are less than
0.1/ug/L.
-------
water being applied. The highest concentration in the water being spray
irrigated was 122 ppb for tnchloroethylene. The highest air concentration
was 9/jg/m3 for trichloroethylene.
, highest air concentrations were obtained inside the scrubber house
at the aeration basins. As shown in Table (51 chloroform was found at a
•eye] of 3380;ug/m3 (approximately 3.4 mg/m3). The results for air samples
obtained at three off-site locations are also shown in Table 61. One of
the off-site locations (lakeshore) had measurable amounts of the chlorinated
compounds. This sample was collected near the paper mill whose discharge
enters MWMS.
. Preliminary studies of air levels of chloroform, trichloroethane,
trichloroethylene and tetrachloroethylene at other wastewater treatment
Jollities indicates that the levels found at the aeration basins of the
Muskegon Treatment facility are comparable to or in some cases less than air
'evels found at these other types of wastewater treatment facilities (Tables
62 and 63).
.... AH air concentrations measured were well below recommended occupational
limits (38). In the event of accidental discharges of large amounts of
these chemicals, areas near the aeration basins, and in particular in the
scrubber house, would be expected to experience the highest air concen-
lrations.
145
-------
TABLE 61. AIR CONCENTRATIONS OF SELECTED CHLORINATED ORGANICS
AT VARIOUS LOCATIONS
Concentration, jug/tn3
Inside Scrubber House
Off-Site Locations;
Road Commission
- Cottage Grove
Lakeshore
tnioroform
3380
NO*
NO
24
irictuoro-
ethane
805
ND
ND
3.1
iricnioro-
ethylene
306
ND
ND
1.9
ietrach loro-
ethylene
240
ND
ND
1.2 __
aND - None detected, concentrations are less than 0.1/jg/m3.
TABLE 62. AIR SAMPLES COLLECTED AT A 30 MILLION GALLONS PER
DAY PRIMARY WASTEWATER TREATMENT PLANT
Concentration, >ug/m3a
Trichloro-c Trichloro-c Tetrachloro-c
Location Chloroform*3 ethane ethylene ethylene
Top of Sed. Basin 0.5 1.0 0.3 1.4
Upwind-NE Fence 1.6 0.4 0.1 0.4
Inside Sludge Dewatering
Bldg. 1.2 5.3 7.5 12.3
V
Enclosed Grit Chamber
(Detroiter Tank) 18 61 10 36
Bar Screen (Wet Well) 53 325 66 145
aData represent single samples at each location.
^Collected on charcoal tubes.
^Collected on Chromosorb 102 tubes at flow rate of 30-50 mL/min.
146
-------
TABLE 63. AIR CONCENTRATIONS OF SELECTED ORGANICS COLLECTED AT
VARIOUS LOCATIONS AT.A 300 MILLION GALLONS PER DAY
ACTIVATED SLUDGE WASTEWATER TREATMENT PLANT
Concentration^g/mS*
Location
Trichloro-c Trichloro-0 Tetrachloro-c
Chloroform'5 ethane ethylene ethylene
Enclosed Aerated
Grit Chamber
Wet Well Pumping
Bldg.
Secondary Aeration
Basin
Inside-Sludge*
Proc. Bldg.
84
47
11
46
20
26
15
15
10
214
139
14
aData represent single samples at each location.
Collected on charcoal tubes.
Collected on Chromosorb 102 tubes at flow rate of 30-50 mL/min.
147
-------
SECTION 10
SERO-EPIDEMIOLOGIC SURVEY OF WORKERS AT A WASTEWATER
-SPRAY IRRIGATION SYSTEM
Spray application, one of the commonly used methods of land appli-.
cation of wastewater, promotes aerosol formation. The viruses present in
wastewater may become airborne during spray application and are thus a
source of exposure to the workers involved. In order to assess the po-
tential health risks to workers from exposure to viruses, virus antibody
levels, isolations and illness rates of the workers at the Muskegon Waste-
water Management System, Michigan,, were evaluated over a 5-month period and
compared with a control group of road commission workers.
METHODS
The populations compared were workers at the Muskegon County Wastewater
Management System No. 1 and the Muskegon County Road Commission in Michigan.
A description of the spray irrigation treatment system has been presented
in Section 7. The Muskegon County Road Commission facilities are located
on the north side of Apple Avenue, east of Maple Island Road (see Figure
5).
Serum specimens and throat and rectal swabs were obtained from the
study volunteers on a monthly basis between June and October 1979. Illness
symptoms experienced during the prior month were obtained during the July-
October visits with the study participants. The form used for this purpose
is shown in Appendix A.
Viral cultures were inoculated into three types of cell cultures,
including primary African green monkey kidney, Vero monkey kidney and WI-3°
cells. These were incubated at 35°C, and examined daily for evidence of
viral cytopathic effect. Viruses, including those recovered from wastewater,
were identified by neutralization with Lim-Benyesh-Melnick antisera pools.
Antibody to enteroviruses was measured by a microneutralization assay.
Iir,riS!S S1ec?«for the survey were P°li°virus types 1, 2 and 3, coxs
II™ K a1d B5' and ech<>virus types 7 and 11. These viruses were
chosen because .hey were isolated from the study population and wastewater
samples, or were the most common enteroviruses in the Muskegon area during
radioiSnoa y t0 hepatitis A virus was measured by a competitive
148
-------
RESULTS
Volunteers Recruited
A total of 35 workers at the wastewater spray irrigation system and 41
at the Road Commission were recruited into the study. The age, race, sex
and length of time on the job for these volunteers is summarized in Table
64. The median age of the Road Commission workers is 10 years higher than
that of the spray irrigation workers. The difference in number of years on
the job reflects the fact that the spray irrigation system began operation
in 1973 with the first workers hired in 1972 whereas the Road Commission
was established in- 1894.
On the basis of interview and job observation data, workers at both
the spray irrigation and road commission facilities were divided into three
relative exposure categories. The spray irrigation workers in the high
exposure category had routine exposure to both wastewater and-aerosols.
Within this group, there was a subgroup of six past and present nozzle
cleaners who were judged to have the highest exposure. Nozzle cleaners are
college-age students hired for the summer vacation months for the purpose
of regularly cleaning nozzles on the spray irrigation rigs and for performing
other tasks. The intermediate exposure spray irrigation system workers had
some exposure to either wastewater or aerosols but not to both. The low
exposure spray irrigation system workers had no exposure to wastewater and
no exposure to aerosols except that which might occur in driving on the
roadway leading to the administration building. This roadway passes about
100 meters from several of the irrigation rigs. The road commission workers'.
were divided into two outdoor 'groups; and one indoor group. The age, race,
sex and length of time on the job for study participants, divided according
to exposure: categ^^
the youngest group, with a median age of 2T'years.
Reported Illnesses
Illness symptoms reported during the monthly interview were tabulated
in the categories: respiratory, gastrointestinal, respiratory plus gastro-
intestinal and other. A comparison of the number of illnesses by illness
type for the spray irrigation and road commission workers, by total group
and by exposure category (Table 66) revealed no differences. The overall
illness rate/worker month was practically identical for the spray irrigation
workers and the road commission workers, 0.54 and 0.58, respectively.
Virus Isolations
Coxsackie B5 virus was isolated from both the throat and rectal swabs
collected in August from one road commission participant. An extensive
interview with this participant failed to detect any evidence of symptoms.
No other isolates were obtained from throat and rectal swabs during the
study.
149
-------
TABLE 64. AGE, RACE, SEX AND LENGTH OF TIME ON THE JOB FOR SPRAY
IRRIGATION SYSTEM AND ROAD COMMISSION STUDY PARTICIPANTS
Spray IrrigationRoad Commissionm
System
Total Number*
Age, years
Median
Range
Race
White*
Non-white*
Length of time on job, years
Range
35(2)
28
19-61
33(2)
2
-------
TABLE 65. AGE, RACE, SEX AND LENGTH OF TIME ON THE JOB FOR STUDY
PARTICIPANTS DIVIDED ACCORDING TO EXPOSURE CATEGORY
" .. -. Spray Irrigation System Road Commission
ExposureJDategory
Total Number***
Age, years
Median
Range
Race
White***
Non-White***
Length of time on job,
years
High*
12
27
19-61
11
1
•«.;
Int.
10
32
24-49
9
1
<,-7
Low
13(2)
27
22-61
13(2)
0
<,-7
Out- "
side •!**
18
34
29-62
15
3
-------
TABLE 66. COMPARISON OF ILLNESS FOR SPRAY IRRIGATION AND ROAD
COMMISSION WORKERS BY ILLNESS TYPE, WORKER GROUP,
AND EXPOSURE CATEGORIES
No. Illnesses
No. Worker Respira- Gastroin- Resp. &
Worker Group Months tory testinal Gast.ro. Other Total
Spray Irrigation 112 33 18 1 9 61
High Exposure 35 10 5 0 4 19
Int. Exposure 39 11 5 0 4 20
Low Exposure 38 12 8 1 1 22
Road Commission 130 52 12 3 8 75
Outdoor-soil
contact 54 23 6 0 4 33
Outdoor-Other 17 10 0 01 I"1
Indoor 59 19 63 3 31
152
-------
Fourteen viral isolates picked from a sample of the July 17, 1979 raw
influent to the aeration basins were identified as follows: two poliovirus
Hi, nine coxsackievirus B2 and three echovirus 7.
Seroloqv
A comparison of the prevalence of antibody for those viruses tested
was made between the spray irrigation and road commission workers for that
month for which the most persons were active in the study, July. The results
of these comparisons are presented in Table 67. Only for Coxsackie 85 was
the difference significant with the spray irrigation worker group having a
Higher proportion of titers^2.
Increases of fourfold or greater in titer level of a particular antibody
(2 or more serum dilutions) indicate infection in the individual involved.
In order to evaluate differences in the number of fourfold or greater titer
level rises results from the first and last serum from each participant
were compared. Comparisons were made for all spray irrigation and road
commission workers. Results, presented in Table 68, reveal no differences
for any of the viruses for either of the groups compared.
The titer level of coxsackie B5 antibody of the workers from whom
coxsackie B5 virus was isolated increased f rom < 2 at the tme of the isolation
to >128 one month later.
Anti-HAV sera from 8 of the 35 spray irrigation workers tested positive
while for the road commission workers the proportion was 17 of 41. A com-
parison of the results divided according to age, *. 30 years, 3 30 years,
MO years and •> 40 years (Table 69) indicate no. significant differences
(chi-square, X^O.IO).
In an effort to further evaluate the effect of age on antibody titer
levels a group of 31 spray irrigation and road commission workers were
selected. This group is composed of 13 low or non-exposed spray irrigation
system workers and the 18 indoor road commission workers. Together their
a9es ranged from 22 to 69. Using Pearson correlation coefficients as well
as X2 and Fisher's exact test, age was found to be directly (and categor-
ically) related to the presence of antibodies to Coxsackie B5 as well as
Hepatitis A. As age increases the probability of finding a "positive (>
2) titer level increases. Exemplary X2 tables are:
Titer Levels to
Cosxackie B5
Test for Antibodies to
Heptatis A
Negative
Positive
Age
<30
> 30
<30
Age
^30
p-UJ.Ol «rby Fisher's exact test -> p<0.005
153
y
8
i
13
IU
10
0
11
-------
TABLE 67." COMPARISON OF PREVALENCE OF ANTIBODY AT TITER LEVEL 2
OR GREATER FOR ALL SPRAY IRRIGATION AND ROAD COMMISSION
WORKERS DURING THE MONTH OF .JULY
liters
SI 1
Negative Positive
Echo 11
Coxsackie B5
Polio I
Polio II
Polio III
Coxsackie B2
Echo 7
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
11
21
11 25
13
24
4
4
0
1
1
3
11
13
27
25
19
12
28
32
32
35
31
33
21
23
5
11
p > .10
P - -03
p > .10
p > . 10
p > . 10
p > • 10
P > . 10
P by X2 test or Fisher's exact test wherever appropriate.
154
-------
TABLE 68. COMPARISON OF NUMBER OF FOURFOLD OR GREATER TITER LEVEL
INCREASES IN SPRAY IRRIGATION AND ROAD COMMISSION
WORKERS USING RESULTS FROM FIRST AND LAST SERUM
COLLECTED FROM EACH PARTICIPANT
_ Virus
Polio I
Polio II
Polio III
Coxsackie B2
Coxsackie B5
Echo 7
Echo IT"
Total
Group
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
Spray Irrigation
Road Commission
No.
Participants
33
40
' 33
40
33
40
33
40
33
40
33
40
33
40,
33
40
No. Fourfold
and Greater Titer
Level Increases
5
5
2
2
3
1
2
5
3
1.
1
1
1
, . o ' ,- ,
17
15
155
-------
TABLE 69. COMPARISON OF HEPATITIS A ANTIBODY TESTS FOR SPRAY
IRRIGATION AND ROAD COMMISION WORKERS
AT BOTH AGE 30 AND AGE 40
Anti-HAV
Spray Irrigation
Road Commission
18
4
3
1
Anti-HAV
Age<40 Negative Positive
Spray Irrigation
toad Commission
23
19
4
7
Age >30
Spray Irrigation
Road Commission
9
20
5.
16
Age>40
Spray Irrigation
Road Commission
4
5
4
10
-------
.._ Using age 30 for partitioning seems crucial in establishing the age
aifference for Coxsackie 85 as not statistically significant (p 0.10) dif-
ference manifests itself if we use age 40. For Hepatitis, however, the
significant difference is unaffected by a choice of age 30 or 40. Although
no significant age effect was statistically discernable for the other viruses
studied (Echoviruses 7 and 11, Coxsackie B2, and Polios I, II and III) they
may, in fact, exist. For our small number of individuals they did not
reveal themselves.
Antibody prevalence comparison between the high exposure spray irrigation
system workers and the indoor road commission workers (Table 70) revealed
no significant differences for the month in which the highest number of
nigh exposure workers were tested (June). In order to evaluate differences
ln the number of fourfold or greater titer level rises, results from the
first and last serum were compared for the high exposure spray irrigation
workers and the indoor road commission workers. Results, presented in
Table 71 reveal no differences for any of the viruses for either of the
groups compared.
Included in the group of 12 participants in the high exposure spray
irrigation workers is a sub-group of nozzle cleaners who are thought to
have the highest exposure. Nozzle cleaners are part-time employees hired
for the summer and are generally college students. Five current and one
former nozzle cleaners were participants in the study. Four of the six had
previously spent at least one summer as a nozzle cleaner, the other two had
°een on the job for a month before the collection of the first specimen for
the study. To obtain statistically significant differences (at say p^O.05)
for only 6 individuals requires a near total polarity in data. This did
occur, in the case of the virus Coxsackie B5. All 6 individuals had positive
(>2) results for all bloods drawn with the minimum titer. being 16. Since
it has previously been shown that age is directly related to the probability
°f an individual having a positive antibody titer to coxsackie B5 and the
nozzle cleaners are aged 19-28 (all under 30), a positive titer would be
expected to occur by chance in only one out of ten persons.
Because one nozzle cleaner quit work after only 1 month of the study
and other summer nozzle cleaners were back in school by September and October
the "optimum" month for comparison was June since that was the only month
when all were in the study. A comparison was made of June Coxsackie B5
titer levels between the nozzle cleaners and various other worker groups:
all road commission workers, outdoor road commission workers with soil
exposure, indoor road commission workers, all spray irrigation workers,
other high exposure spray irrigation workers, other high exposure spray
irrigation workers and intermediate exposure spray irrigation workers, and
low exposure spray irrigation workers. For all these comparisons the
significance levels, determined by Fisher's exact test were: £ 0.05 (Table
?2). All 6 of tlie past and present nozzle cleaners showed negative results
for anti-HAV of the remaining 6 high exposure spray irrigation workers,
only 2 individuals were positive.
157
-------
TABLE 70. COMPARISON OF PREVALENCE OF ANTIBODY AT TITER LEVEL 2
OR GREATER FOR HIGH EXPOSURE SPRAY IRRIGATION WORKERS AND INDOOR
ROAD COMMISSION WORKERS DURING MONTH OF JUNE
— —
Titer:
<2
Echo 11 Spray Irrigation 2
High Exposure
Coxsackie B5
Polio I
Polio II
Polio III
Coxsackie B2
Echo 7
Road Commission 5
Indoor Workers
Spray Irrigation 4
High Exposure
Road Commission 8
Indoor Workers
Spray Irrigation 3
High Exposure
Road Commission 2
Indoor Workers
Spray Irrigation 0
High Exposure
Road Commission 1
Indoor Workers
Spray Irrigation 1
High Exposure
Road Commission 1
Indoor Workers
Spray Irrigation 3
High Exposure
Road Commission 8
Indoor Workers
Spray Irrigation 11
High Exposure
Road Commission 10
Indoor Workers
^2
10
D>0.10
9
8
P>0.10
6
9
p>0.10
12
12
p>0.10
13
11
p>0.10
12
1 9
D>9.10
I 6
1
piO.10
I 4
P by Fisher's exact test.
158
-------
TABLE 71. 'COMPARISON OF NUMBER OF FOURFOLD OR GREATER TITER LEVEL
INCREASES IN HIGH EXPOSURE SPRAY IRRIGATION WORKERS AND
INDOOR ROAD COMMISSION WORKERS USING RESULTS FROM FIRST
AND LAST SERUM" COLLECTED FROM EACH PARTICIPANT
Virus
Polio I
Polio II
Polio III
Coxsackie B2
Coxsackie B5
Echo 7
Echo 11
Cumulative
Group
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
High Exposure Spray
Irrigation
Indoor Road Commission
No.
Participants
11
17
11
17
11
17
11
17
11
17
11
17
11
17
1^
1
17
No. Fourfold and
Greater Titer Level
Increases
2
2
0
0
1
1
2
2
'.'".•• • -
1'
•-..•:;
1
0'
0
7
/
7
159
-------
TABLE 72. COMPARISON OF JUNE COXSACK1E B5 TITER LEVELS FOR NOZZLE
CLEANERS WITH THOSE OF VARIOUS OTHER GROUPS
OF STUDY PARTICIPANTS
June
Coxsackie B5
Tilers
<2 »2
Nozzle cleaners
Outdoor Road
Commission Worker
Workers-soil
contact
Negative
0
' 9
Positive
6
6
June
Coxsackie B5
liters
42 >2
Nozzle cleaners
Other Spray
Irrigation High
Exposure
Negative
0
4
Positive
6
2
0.017
0.030
Nozzle cleaners
Road Commission
Indoor Workers
0
8
6
6
0.024
Nozzle cleaners
Spray Irrigation
Low Exposure
0
7
6
2
0.006
Nozzle cleaners
All Road Com-
mision
0
19
6
15
0.014
Nozzle cleaners • 0
,i
Spray Irrigation
Other High Ex-
posure and Int.
Exposure
9
6
6
P ' O.Q17
Nozzle cleaners
All Other Spray
Irrigation
Workers
0
16
6
8
Critical levels (p) are determined by Fisher's exact test.
160
-------
It is apparent that further investigation is warranted in the case of
the nozzle cleaner category of workers. These individuals are reported to
become frequently soaked when back pressure remains in the nozzles. It is
also likely that infection with the virus may occur symptom-free, as was
the case with a road commission worker whose titer level increased from
to >_]28 in one month from August to September with confirming throat and
rectal swab tests but with no noticeable symptoms.
No differences in illness and virus isolation rates were detected be-
tween the 35 spray irrigation workers and the 41 road commission workers.
The only virus isolated by routine screening was Coxsackie B5 which was
identified in both the throat and rectal swab of an asymptomatic road
commission .worker whose titer rose from less than 2 to greater than or
equal to 128. The spray irrigation workers had elevated levels of antibody
to coxsackvirus B5 compared to the control group of road commission workers.
Levels of antibody to Coxsackie B5 in a group of six present and former
nozzle cleaners were significantly higher than for the other spray irrigation
workers, the road commission workers or for several subgroups based on
relative exposure. This difference was not due to age differences between
the groups, because although the prevalence of the virus tended to increase
with age, the spray irrigation workers were, on average, younger than the
road commission workers. No differences were detected in levels of antibody
to Poliovirus 1, 2 and 3, Coxsackie B2, Echo 7 and Echo 11. Presence of
Hepatitis A virus was found to be positively correlated with age, as has
been found by others (2,_n4,jl5} but there were no differences between
groups when corrected for age.
161
-------
•SECTION 11
EFFECTS OF SEWAGE TREATMENT PLANT SLUDGE ON THE STRUCTURE
AND FUNCTION OF TWO GRASSLAND ECOSYSTEMS
Numerous studies have been concerned with the effects of sludge on the
cellular (116), histological (117), and organismic (118) levels of biological
organization. Most of these studies have focused upon individual plant or
animal species or their respective component parts (e.g., concentration of
heavy metals within various tissues and organs or effects of certain sub-
stances upon individual growth and/or reproductive rates). However, since
all species including man, function within natural ecological systems (eco-
systems), it is unfortunate that the higher levels of biological organi-
zation (i.e., the population, community, and ecosystem levels of integration)
have received little attention.
Odum, Barrett, and Pulliam (119) have pointed out that problems often
arise when stressors are tested at one level (e.g., individual) and then
used without further study at another level (e.g., ecosystem). It is also
clear that only by considering the whole ecosystem, as outlined in this
report, will man be better able to predict the long-range results of environ-
mental contamination (120). Further, Barrett, Van Dyne, and Odum (120) have
outlined specific guidelines for testing stressors at the ecosystem level.
We believe "field ecosystem testing" is not only feasible but may also be
much -less expensive and more realistic than laboratory tests of each species
or population separately. It is an ecological axiom that the sum of the
parts does not equal the whole (122).
Woodwell (123) challenged engineers and planners to convert cities from
open systems which take in fresh water and excrete wastewater to closed
systems which reuse their water resources. A possible means of accomplishing
this goal is to filter sewage wastes through managed ecosystems. Effective
planning should locate such systems in the vicinities of municipalities
according to the compartmentalization concept of Odum (124). Indeed, the
feasibility of this approach regarding forest and cropland ecosystems has
already been tested (125). The "best compartments," (i.e., the most efficient
ecosystems) may be an intermediate (natural) old-field plant community in
the 3-10 year of old-field succession which would naturally decompose and
assimilate the greatest amount of sludge on a per-unit (ha) basis. The
reasons for this are related to Table 73. Old-field communities are highly
productive (126), exhibit closed inorganic nutrient cycles (127), make ef-
fective use of intrabiotic detritus (128), and are considered to be eco-
logically stable (129). Such early developmental communities could also
handle additional nutrient input (extrabiotic input) and still maintain the
162
-------
TA3LE 73. ATTRIBUTES OF THREE STAGES OF ECOSYSTEM DEVELOPMENT
RATED ACCORDING TO THEIR EXPECTED CAPACITY FOR PROCFSSIN6
SEWAGE SLUDGE (Adapted From Odum, 1969) (124)
1.
z.
3.
4.
5.
6.
7.
8.
Ecosystem Attributes
Community energetics
Gross production/standing
crop biomass
Net community production
Community structure
Inorganic nutrients
Life history
Life cycles
Nutrient cycling .
Mineral cycles
Nutrient exchange rate
Role of detritus
Overall homeostasis
Stability
Young (Wheat)
Agricultural
Stage
High
High
Extrabiotic
Simple
Open
Rapid,
Unimportant
Poor
Intermediate*
(Old-field)
Stage
High
High
Intrabiotic
Intermediate
Closed
Rapid
Important
Good
Mature
(Forest)
Stage
Low
Low
Intrabiotic
Complex
Closed
Slow
Important
Good
*proposed most efficient ecosystem for sewage disposal,
163
-------
above-mentioned characteristics. In addition, the energy subsidies which
operate or drive these systems should be much less as compared to either
agricultural or forested systems which frequently require human (energy
intensive) management. The producers (plants) are naturally seeded and
maintained and the primary consumers, dominated by arthropods, are natural
grazers containing high protein content for higher trophic levels. By intro-
ducing meadow voles (Microtus pennsylvanicus) into these communities the
rate of nutrient recycling on a low energy basis would be greatly increased.
The voles are effective plant consumers |30 ) and would also be important
warm-blooded monitors concerning growth, reproduction, and population
dynamics change.
The objectives of this study were as follows: (a) to measure and evaluate
the effects of chronic sludge application on several different levels of
biological organization, (b) to compare two different community-types (mono-
culture vs. old-field) treated in an identical manner, (c) to analyze dif-
ferent structural (e.g., biomass and species diversity) and functional (e.g..
energy flow and community resilience) ecosystem parameters—measurements
which may be indicative of total system response, (d) to evaluate sludge
effects (via ecosystem food chains) on a natural small mammal population
(Microtus pennsylvanicus) functioning within these community-types, and (e)
to develop a feasible ecosystem approach, including the identification of
"white mice" field parameters, for testing stressors (e.g., sludge) on intact
ecosystems.
METHODS
Land Treatment
The study made use of landspreading techniques to examine the effects of
chronic dosages of sludge to compare an old-field plant community with an
agricultural (wheat) field community. The experimental system consisted of
sixteen one-tenth hectare enclosures of galvanized steel located at the
Ecology Research Center at Miami University. The individual plots were
randomly selected for treatment with sludge (S), fertilizer (F) or were used
as controls (C) as indicated in Figure 22. The sludge used was Milorganite,
an anaerobically-digested, heat-dried municipal sludge commercially marketed
by the City of Milwaukee, Wisconsin. The sludge was applied to each of a
total of six plots at the rate of 181 Kg per month during May-September in
1978 and 1979. Its elemental composition was 6-2-0 (N-P-K).
Commercial fertilizer of composition (34.2-11.3-0) was applied at the
rate of 32 Kg per plot per month to provide equivalent added amounts of
nitrogen (N) and phosphorous (P) as received by the sludge amended plots.
Eight of the plots were planted in Kentucky Bluegrass (Poa sp_) in 1974 and
were in the fourth year of old-field succession in 1978"The other eight
were planted in winter wheat in fall of 1977. The wheat matured in July
1978 was not harvested, and was left fallow thereafter. The wheat field had
previously been treated as follows:
164
-------
o
CO
LL=m
Um
CO
1
CO
o
U=m
CO
CO
o
o
Urn
CO
"."-•'.:
Li-
ca
S: Sludge
F: Fertilizer
C: Control
Figure 22. Schematic arrangement of individual study plots,
165
-------
20 October 1977--plowed and disked;
24 October 1977—fertilized with 300 Ibs/acre (336 Kg/ha) commercial
fertilizer (12-12-12) and seeded with winter wheat
(Triticum sp.).
A modified garden manure-spreader was used to dispense the dried sludge and
fertilizer.
Contamination of control plots by sludge from adjacent experimental
plots was minimal and was restricted to the lower soil horizons. Steel en-
closure walls surrounding the plots extended 20 inches (60 cm) below the
ground surface, thus restricting any lateral flow of nutrients. The only
possible source of contamination was through deep root extraction of nutrients
leached to lower soil layers.
Sludge and Fertilizer Analyses
Composite samples were made for each month of sludge and/or fertilizer
application for 1978 and 1979. Five gram portions from subsamples of the
same month were combined and shaken vigorously (six plots, two bags per
plot). This composite, which was felt to be representative of the month was
then analyzed for Pb, Cd, Cu and Zn in triplicate using 1 gm samples. There
were no sludge samples received for the months of June 1979, and there was
only an annual composite fertilizer sample for 1979. In this latter case
six replicate 1 gm samples were taken.
Sample preparation for analysis by flame atomic absorption spectroscopy
involved the use of a wet-ash technique using nitric (HNOs) and perchloric
(HC104) acids. Five ml of distilled concentrated nitric acid was added to
the sample and the sample was heated gently to dryness. After cooling 10 mL
HMOs and 10 ml HClfy were added. The samples were then heated to fumes of
HC104. After cooling, the samples were retreated with additional 5 ml HN03.
The samples were again heated and this time allowed to go to dryness. Addi-
tional treatment was felt necessary, especially for the sludge samples;
consequently, 5 ml portions of each acid were added, and the samples were
reheated strongly to dryness. Final dissolution consisted of adding 10 ml
of acid solution 5% (v/v) nitric acid and 5% (v/v) hydrochloric acid in
demineralized H20, concentrating to approximately 2 ml, transferring to 10
ml graduated cylinder, making fresh up to the original volume, and shaking
vigorously. The undissolved material was allowed to settle out prior to
analysis. Reagent controls were also carried with the samples.
Ecological Methods
Five breeding pairs of meadow voles (Microtus pennsylvanicus) were
IX™° T! 1nt° each of the 16 quarter-acre plots during June of 1978 and
is/9. The total of 320 voles was captured from wild (untreated) populations
in nearby hayfields. iff
ma« KThe^tUJy/uCusfd on three asPects of the grassland ecosystems which
may be affected by sludge deposition: structure and energetics of the total
166
-------
ecosystems, population dynamics of small mammals, and population dynamics of
arthropods.
Comparison between control and experimental plots in each grassland
type provided information regarding the effects of sludge on the ecosystem
as a whole. Comparison of the experimental plots between the two grassland
types provided information as to which ecosystem is better able to process
sludge most efficiently.
The effects of sludge on vegetative community structure was considered
°y comparison of species diversity between control and experimental plots.
^oductivity was measured by the short term harvest method. Diversity was
Determined by the number of species per 0.25 m~2, Margalef's richness index,
Cannon-Weaver's function, and Pielou's evenness index.
Vole population dynamics in the control and experimental systems were
compared on the basis of age structure, turn-over rate, population growth
rate, and carrying capacity. Vole population densities were related to
vegetation nutrient content in a test of Pitelka and Shultz (131) theories
Of nutrient-recovery and nutrient cycling effects.
Arthropod response to treatment was evaluated for all major orders in
terms of density and biomass. One order (Homoptera) was analyzed in detail
regarding species richness and apportionment.
During November of 1978 and 1979 the voles were caught in the morning,
numbered, and delivered to the Pathology Section in the Department of Environ-
mental Health at the University of Cincinnati for sacrifice and autopsy.
tach day the animals were randomly selected from different fields. A complete
autopsy was performed on each vole. Tissues were taken from each organ and
Jixed in neutral formalin. Histopathology was performed on liver, lung,
kidneys, and testes or ovaries. The remaining organs were stored in the
fixative in case more extensive examinations are desired.
The tissues were embedded in paraffin, cut at 6 urn thickness, and stained
yith hematoxylin and eosin. They were histologically examined without know-
ledge of the treatment. The findings were recorded on the autopsy records.
After completion, the code of the different groups was received and the
Pathology was tabulated.
RESULTS
Content of Sludge and Fertilizer
The-results of the analysis of the heat-dried sludge (Milorganite) and
tne commercial fertilizer for lead, cadmium, copper and zinc are presented
in Tables 74 and 75 respectively. For each metal the concentration in the
sludge was higher than in the fertilizer. The ratio of the concentration in
the sludge to the concentration in the fertilizer in 1978 was 88 for Pb, 30
167
-------
TABLE 74. METAL CONTENT OF SLUDGE, 1978 AND 1979
1978
Concentration,jug/g, mean
Pb Cd Cu
May
June
July
August
September
May- September
448 + 10
487 + 4
488 + 13
476 + 10
467 + 5
473 + 17
60 + 1
59 + 1
60 + 2
59 + 1
60 + 1
59 + 1
»
341 + 4
325 + 5
308 + 7
313 + 6
315 + 4
320 + 13
+ S.D.a
Zn
1012 + 20
1088 + 12
1 162 + • 80
1075 +13
1062 + 93
1081 + 67
1979 .
Concentration, /ig /g, mean +_ S.D.
May
dune
July
August
September
May-September
Pb
454 +
Cd
14
No samples
449 +
467 +
466 +
459 +
43
18
28
25
60+1
No
57
57
57
58
samples
+ 5
± 3
+ 2
+ 3
Cu
341 +
2
No samples
313
330
331
329
+
+
+
+
38
8
16
21
Zn
1007 +
28
No samples
926 +
959 +
978 +
968 +
144
22
38
72
aThe values represent triplicate determinations.
168
-------
TABLE 75. METAL CONTENT OF FERTILIZER, 1978 AND 1979
19783
May
June
July
August
September
1978
Concentration, jug/g, Mean + S.D.
Pb Cd Cu Zn
1.8+0.2 1.6+0.1 1.4+0.0 20.8+0.4
5.7+7.0 2.2+0.2 1.5+0.1 19.5+0.7
3.0+0.8 2.0+0.1 2.6+0.7 26.8+1.6
4.9 + 6.8 2.3 + 0.8 2.2 +_ 0.4 24.5 + 6.4
10.3+15 1.7+0.1 1.4+0.2 16.7+0.4
May-September 5.4 + 7.6 2.0 + 0.4 1.9+0.6 21.8 ±4.6
Annual
Composite 12.4+12.0 1.6 ±0.4 1.6 + 0.4 16.4 + 4.0
Triplicate determinations.
be-
i-|x replicate determinations.
169
-------
for Cd, 168 for Cu and 50 for Zn; for 1979 the ratios were were 37, 36, 205
and 59, respectively. Since the rate of sludge application, by weight, was
5.6 times that of the fertilizer, appreciably more of these metals were
applied to the sludge-treated plots than was indicated by the concentration
ratios alone as shown in Table 76.
Primary Productivity and Diversity
Shannon-Weaver's function and evenness in the annual field were signifi-
cantly lower in the fertilizer than in the sludge treatments (Table 77 and
Figure 23) (132,133). No significant differences due to treatment were
found in species richness in the annual field or in any diversity index in
the perennial field (Table 77 and Figure 24).
There was a significant increase in productivity in both fields in the
fertilizer and sludge treatments, however the increase was larger and occurrea
earlier in the fertilizer treatments (Tables 78 to 81 and Figures 25 and
26). Estimates of net primary productivity in the annual field were 1690 g"
2 yr"1 + 84 S.E. in the fertilizer treatment, 1240 g-2 yr"1 + 58 S.E. in the
sludge treatment, and 772 g~2 yr'1 + 3 S.E. in the control. Estimates of
net primary productivity in the perennial field were 1110 g'2 yr'1 + 39 S.t.
in the fertilizer treatment, 684 g'2 yr'1 + 69 S.E. in the sludge treatment,
and 424 g"2 yr-1 +_ 37 S.E. in the control. It appears that the treatment
effect on plant community structure was greater in the annual field than in
the perennial field, suggesting the perennial community is more stable than
the annual field and better suited for application of sludge than the annual
field.
Arthropods
Arthropod density and biomass increased in both communities following
treatment with fertilizer or sludge (134). The Coleoptera, Diptera, Hemiptera
and Homoptera depicted total density and biomass arthropod response to nutrient
enrichment in both communities. Significant density differences occurred
later in the growing season in the old-field plots as compared to the agri-
cultural plots; the agricultural plots exhibited greater resilience to
nutrient perturbation whereas the old-field plots exhibited greater resistance.
Diptera, Hemiptera, and Homoptera in the old-field community exhibited
significant increases in biomass as a result of nutrient enrichment. All
orders except the Homoptera and Araneida in the agricultural community showed
significant increases in biomass as a result of nutrient enrichment. Homoptera
and Araneida densities were signficantly greater in the old-field plots
whereas Hemiptera densities were greater in the agricultural plots. Also,
all orders except the Hemiptera and Araneida showed greater biomass values
in the old-field as compared to the agricultural community.
The Homoptera, a dominant group of insect herbivores, was used for an
analysis of species/unit area, species richness (D = S - 1/ln N) and species
apportionment (e = H'/ln S). Fertilizer and sludge plots exhibited con-
sistently higher species richness values than did the control plots in both
170
-------
TABLE 76. TOTAL AMOUNTS OF THE HEAVY METALS APPLIED TO THE SLUDGE AND
FERTILIZER TREATED PLOTS IN 1978 AND 1979 (GRAMS)
Sludge
Metal
Lead
Cadmium
Copper
Zinc
1978
2568.
320,
1737
'5869
39
.37
.60
.83
1979
2492.
314.
1786
5256
Fertilizer
1978
37
,94
.5 :
.24
5.
1.
1.
20
18
92
82
.93
Sludge/Fertilizer
1979
11.
1.
1
15
9
,53
.53
.74
1978
495
167
953
280
Sludge/Fertilizer
1979
209
205
1163
334
-------
TABLE 77 SHANNON WEAVER'S DIVERSITY INDEX, MARGELEF'S RICHNESS INDEX, AND PIELOU'S EVENESS INDEX
IN THE ANNUAL AND PERENNIAL FIELD BASED ON PEAK BIOMASS OF EACH SPECIES
ro
Annual Field
Fertilizer
Sludge
Control
Shannon Weaver's index Mar^elef a richness
1.29+0.016' 3.1910.239
1.61+0.237 3-04+0. 39*»
1.59+0.080 , 2.93*0.56
Pielou'e evenness
0.40+0.006 .
0.52+0.055
0.51+0.012
Perennial Field
Fertilizer
Sludge
Control
1.66+0.080
1.61+0.097
1.63*0.125
1.07+0.217
2.3^+0.681
1.82+0.202
0.5^+0.081
0.59+0.075
0.67*0.060
-------
Ul
(O
c
rt)
IS}
co
1.2 5 _
co
31 £=
»* Z
I | 1.0
ro1 m
(i) ™^-i
Q.
(D
X
3 O
0.75-
Oi
c
Oi
a>
Qi
53
3
C
OI
O)
n>
Q.
0.5 _
0.25..
0.0
FERTILIZER-
SLUDGE _
CONTROL
\
4/2B
5/26 6/16 7/7
DATE
7 ;!
8/18
9/15 10/6
-------
(£>
-t
O)
ro
GO
fa
3
O
3
I
CD
0»
-J
«/>
3
a.
a>
x
3 I
-s
ro
CK
-S
o»
in
1.25-
g 1.0
CD
m
0.75-
3 °'s
-a
n>
n>
3
2 0.25,
-s
a>
• 0.0
FERTILIZER
SLUDGE
CONTROL
9/16 10/7
-------
TMLE P™L
Live blotnasg
Annual Field
Fertiliser
Sludge
Control
Perennial Field
Fertilizer
Sludge
Control
938
613
396
(8
(8
(7
533 (7
18)
18)
7)
19)
1*2? (7 19 )
230 (8 19)
4
Litter
25U
131
117
211
(9 15)
(10
(10
(6
232 (7
309 (7
6)
6)
17)
8)
29)
.
Standing dead
507
167
1?8
316
(10 6)
(9 15)
(10 6)
(^ 29)
333 (»* 29)
' ('* 29)
Productivity
21 (6
16 (8
8 (6
16 (8
16)
18)
16)
19)
1U (8 19)
7 (8 19)
-------
TABLE 79. PEAK BIOMASS OF SPECIES WITH A PEAK GREATER THAN 5 q m'
IN THE ANNUAL FIELD. VALUES ARE EXPRESSED AS g rrr2 YR'1. VALUES
IN THE FERTILIZER AND SLUDGE TREATMENTS ARE MEANS OF THREE
REPLICATE PLOTS AND THE CONTROL IS MEAN OF TWO PLOTS
Ambrosia artemieiifolia
Triticuffi aestivum
Chenopodium album
Cirsiura arvense
Setaria laberjj.
Trlfoliun pratense
Polygomm pennsylvanicua
Phlevm Tiratense
Dactylie ploraerata
Me 11 lotus officina.lis
Tri folium hybridum
30 others
Total
Fertilizer
877-88
536^6^
56-26
48±Ji3
21^26
l^f-9
11^7
10-13
at ip
5*-2
1^1
100
1690-146
Sludge
456+U2
326^25
5*-7
52^28
153-83
38^15
10-7
0
8tn
18^8
15-6
1?9
1240-101
Control
151*20
364-1
6*-5
89-69
8t3
110-40
0
0
1-1
26-9
5-2
12
772-5
176
-------
TABLE 80. PEAK BIOMASS OF SPECIES WITH PEAK GREATER THAN 5 g m-2 IN
THE PERENNIAL FIELD. VALUES ARE EXPRESSED AS g nr2 YR"1. VALUES
IN THE FERTILIZER AND SLUDGE TREATMENTS ARE MEANS OF THREE
REPLICATE PLOTS AND THE CONTROL IS A MEAN OF TWO REPLICATE
PLOTS
Species
Zestuca elatior
Zhalaris arundinaceae
?oa pra tense
Solidaeo gpp.
Pactylis g lorn era ta
Meliletus officinalis
ghleua pra tense
Setari a faberii
A^lopereuK pratensis
1? others
Fertilizer
250-54
270*85
120-14
120-91
105-86
55-78
15-12
llt-14
62
Sludge
177*16
200-47
100-27
112^22
48-32
2^2
6^6
13217
13^3
14
Control
125^18
119-4
75^5
2ltl3
• 29-21 .
9^5
-
-
2-1
44
Total
1110-67
684-119
424-52
177
-------
TABLE 81. MEAN + S.E. FOR GRAIN PRODUCTION IN THE WHEAT FIELD
Seeds nT
Heads m~2
Weight (g m"2)
Seeds head"
Seed weieht
Fertilizer
+
1238*110
600^53
18-11
0.04^0.09
Sludee
14980^1491
864^179
426-48
18^11
0.03-0.002
Control __
1140-1077
780-110
288-28
16-13
0.03-0.001
178
-------
to
c
-s
n>
ro
en
-a
-s
o
o.
o
25
O
CD
C
O
2 20
VO
rt
ro
Ol
£
ro
o>
n>
ex
15.
M
Q.
O
10
3
C
CD
(U
(D
Q.
FERTILIZER
SLUDGE
CONTROL
I
4/28
\
' \
/ \
/ \ '.
1 .\ /
// \/'x
/^f
/ •'/ ^\
// ^^_
y \-
\
'^
y \\
\
\
~ * «.
I
\v
II I
5/28 8/18 7/7
7/28 8/18
9/15
DATE
-------
00
O
C
o
CL
n
r+
<'
-J.
r+
<<
«J.
3
(D
•a
a>
ro
(O
-5
CU
fD
a.
O
a
CO
M
0.
-5
tt>
3
3
CT>
-5
fD
CL
20 -
16
12
~ 8
FERTILIZER —
SLUDGE —
CONTROL —
4/29
T
T
5/27 6/17
T
T
7/8
DATE
7/29 8/19
-n 1
9/16 10/7
-------
community types. The opposite was true with respect to apportionment values.
pathology Report on Voles
A total of 16 experimental groups were brought to the laboratory for
examination. Table 82 gives the summary of treatment.
TABLE 82. EXPERIMENTAL GROUPS OF VOLES AND THEIR EXPOSURE
Treatment'Groups
Fertilized agricultural field 01, 02, 04
Sludge agricultural field 03, 07, 08
Control agricultural field 05, 06
Fertilized fallow field - 09, 13, 14
Sludge fallow field 11, 12, 15
.Control fallow field 10, 16
—————— •
No pathologic alterations were seen in the lungs and kidneys of the
voles -- controls as well as treated (135). The ovaries were also of usual
structure. However, some of the animals in each group showed ovaries that
had not fully matured, whereas, others showed recent or. recent and old corpora
futea. A similar situation was present in the males. Some testes were
matured, to different degrees. Since the maturation of the gonads reflects
the age of the animals, the length of exposure was hard to assess. This was
supported to some extent by the histologic findings in the livers, the organ
that had cytologic alterations attributable to exposure.
The least severe hepatic change is depicted in Figure 27. The cells
were swollen and contained multiple vacuoles. These were filled with a
|luid. Their size varied to some extent. The nuclei showed no alterations.
Ih1s was the most common lesion. It is a hydropic degeneration which is
reversible.
. ' A combination of hydropic vacuoles and larger sharply delineated ones
"•s demonstrated in Figure 28. The latter vacuoles appear to be empty; they
actually contain lipid material. Again, the hepatic cells are swollen. The
boundaries between the cells are obscured. This type of degeneration is
more severe and not easily reversible.
The most severe alterations are presented in Figure 29. The hepatic
ceils are ill-defined. Hydropic and fatty changes are seen. Additionally,
some nuclei are small and pyknotic, whereas, others are broken up in pieces.-
lne nuclear membrane is irregular in outline.
, The hepatic changes are summarized in Table 83 for those groups of
v°'es that had been living on agricultural fields. The groups were combined
181
-------
deger,
-------
«•"**
*^t_
^ 1* -^ 4
*«K a *2
*v <•
.
, •
-
Figure 29.
More'detail in' text7 ^Hewtoxylin and
eosin; 250x.
183
-------
since no substantial differences could be detected. The total number of
males and females are given. There seems to be no significant difference in
the incidence of lesions between fertilized and sludge fields. Generally
TABLE 83. LIVER DEGENERATION IN VOLES ON AGRICULTURAL FIELDS
Treatment
Fertilized
Sludge
Control
Number
Males
22
18
13
of Voles
Females
15
18
11
Hydropic Fatty
(f
11
10
4
$ o* £
8 1
9 3
3 1
Nuclear
,7
6
2
-
£
2
2
the alterations are of mild degree. A definite effect is present in the
exposed groups over the control. The more severe changes are not present in
the untreated voles, especially when one considers that the fatty degeneration
occurred in a male without testes.
Table 84 contains the data on voles maintained on fallow fields. Most
frequent was the hydropic degeneration. The incidence seemed to be slightly
higher in the female sludge groups. The control groups had a significantly
lower occurrence of degeneration.
TABLE 84. LIVER DEGENERATION IN VOLES ON FALLOW FIELDS
Treatment
Fertilized
Sludge
Control
Number
Males
19
18
15
of Voles
Females
17
18
6
Hydropic Fatty
cf £ d* £
14
13
4
7 -
11
2 -
Nuclear
d* ?
3 1
2 - ____-
-
Comparing the agricultural groups with the fallow ones, there was a
lower incidence of severe (fatty and nuclear) cell degenerations in the
fertilized fallow field animals (4 degenerations in 36 voles) than in the
fertilized agricultural field ones (9 degenerations in 37 voles). Voles on
the sludge treated agricultural fields (7 degenerations in 36 voles).show a
more severe degeneration than on the sludge-treated fallow fields (no de-
generations in 36 voles). One has to conclude that the agricultural fields
are slightly more hazardous than the fallow fields to the'voles. This,
however, should be considered with caution because the endpoint of toxicity
was measured only on the pathology in the livers. A more sound evaluation
could have been done on the basis of pathohistologic examination of all
organs.
184
-------
Meadow Vole Populations
Meadow vole (Microtus pennsylvanicus) populations were monitored two
years by live-trapping (136). Vole population densities were affected by
treatment only in the first year wheat fields, where sludge treatments had
higher densities than any of the other treatments in the two years (Figures 30
and^l). The fertilizer treatments in the first year wheat fields had low den-
sities due to poor habitat quality as a result of domination of the plant
community by ragweed. In the wheat field vole populations responded to changes
""i plant community structure induced by fertilization, but snowed no apparent
negative reaction to the sludge £er jse. The data points shown are the means of
the treatment replicates on each trapping date. The sludge plots were
significantly higher than the fertilized plots and controls after 21 August
(Duncan's multiple range test at the 95% level of significance). After 4
September all three treatments were significantly different from each other.
Although the fertilized plots had the highest Net Primary Productivity (NPP)
they were so dominated by ragweed after the wheat had matured that much of the
ground cover was shaded out, resulting in poor vole habitat. Ragweed made up
52% of NPP in the fertilized plots, and no other species besides wheat
comprised greater than 3% of NPP. These plots had the lowest Shannon-Weaver
diversity of the three treatments. In the sludge plots ragweed comprised only
34% of NPP, and foxtail and red and sweet clover were relatively productive as
9round cover. Ragweed comprised 15% of NPP in the control plots. Vole
Populations were highest in the sludge plots because the fertilizing effect of
the sludge increased plant production, but at a slower rate than the
fertilizer, allowing lower-growing species to become established and compete
with the ragweed. Survivorship, longevity, percentage of breeding adults, and
recruitment rates were not affected by sludge treatment. Adult sex ratios
favored females in nutrient-subsidized treatments and males in controls. This
'.appears to have been related to .differences in food quality. Juvenile sex
ratios were equal.
There were no treatment differences in tissue concentration of Pb, Zn, or
Cu (135). Cd concentrations were elevated in kidneys and livers of all sludge-
treated voles. The mean liver-and kidney cadmium concentrations in voles from
the control, fertilized and sludge-treated fields for 1978 and 1979 are shown in
Tables 85 and 86. The differences between males and females, and between age
groups are also included in Tables 85 and 86. Cd concentrations were high in
the kidneys of all sludge-treated voles regardless of community-type. Al-
though vole population dynamics were not adversely affected, there exists the
Possibility for long-term, low-dose Cd effects in sludge-treated plots. No
organ or whole weight differences were noted between treatments. There was
slight liver pathology in voles from both nutrient-subsidized treatments.
This may have been due to social stress and not directly related to treatment.
No treatment differences were observed in learning ability or memory as
Measured by the swim-escape test. The mid-successional old-fields appear to be
Preferable to the early stages of succession or to the agricultural fields as
land application sites for sludge disposal in terms of minimizing effects on
small mammal populations.
In the old-field the plant species showed no differences in diversity or
composition between treatments as was previously discussed. Vole populations
185
-------
00
en
Number
Present
40-
35-
30-
25-
20-
15-
-./
^ • * s^r AA^
( VI— -»^ /•
10 5 14 21 2 16 25 1 16
Jun Jul Aug Sept
w ^
•'
A--x.^
••••*.—•
sludge
fertiiizea
control
23 7 18 30
Oct
1978
Figure 30. Meadow vole population densities in the wheat field.
-------
20-
Old Field
oo Number
Present
sludge
fertilized
control
10
Jun
Jul
20 10 22 5 15
Aug Sept
1978
3 10
Oct
30
Figure 31. Meadow vole population densities in the old-field,
-------
CO
00
TABLE 85. CADMIUM CONCENTRATIONS* IN VOLES FROM THE WHEAT FIELD (1978) AND FIRST-YEAR
OLD-FIELD (1979) (jug/g WET WEIGHT)
,
Liver
adult males
adult females
sub.adult males
cub. Add It females
K Idiwy.t
adult unties
«dnlt fnnalrs
sub. adult males
sub. adult frmnlca
*Means of treatment
Control
1978
0.01 (3) e
0.0« (1) e
n.Ol ('I) e
0.02 (3) e
0.01 (1) f
0.20 (2) f
o.oi (2) r
o.os (?) r
replicates
OrtUlKCil
1979
0.03 (3) e
0.06 (?) de
0.02 (3) e
O.OM (2) e
0.16 (2) f
0. 59 (?) ff
O.I? (?) f
0.11 (I) f
: the number
19 7»
(1.03 (5) e
0.09 (5) lif
0.03 (G) e
O.OM (i|) e
0. r
O.OT (?) f
of samples
1979
O.C'i (3) f
a. 09 CO >if
0.05 (5) e
O.OU (3) e
o.3i (?) r
O.RB (3) ilef
o.ro (3) r
n.i7 (i) r
Sludge
1978
0. 1'l (•!) ctle
0.5G (5) b
0.20 (5) cde
0.29 (G) c
0.90 (3) drf
2.10 (3) be
0.03 (2) of
1.37 (3) nln
is in parentheses; means
1979
0.28 (<1) cd
0.90 ('I) a
O.'l7 ('!) «
0.83 CO n
1.72 (3) lied
3,50 (3) n
1.71 (3) brd
2. 3'l (3) b
within
each organ followed by the same letter are not significantly different, between different
age-sex groups and years, and among the three treatment fields (p>0.05, Duncan's multiple
range test).
-------
CO
vO
TABLE 86 CADMIUM CONCENTRATIONS* IN VOLES FROM THE FOURTH-YEAR (1978) AND FIFTH-YEAR
(1979) OLD-FIELDS (jjg/g WET WEIGHT)
, — _ — , — ' - —
Liver
adult males
adult females
sub. adult males
sub-adult female*
Kidneys
adult males
adult females
sub-adult males
sub-adult females
*Means of treatn
Control
0.02 (1) d
0.26 (1) lied
O.ll (3) cd
O.l«l CO cd
0.15 (2) e
-
0.15 (2) e
0.70 (2) cde
lent replicates
lowed by the sai
1979
O.O'I (2) cd
O.'l'l (3) b
0.02 (3) d
O.O'I (3) cd
O.'IO (2) de
1.61 (2) bcde
0.01 (2) e
O.JB (2) e
; the number
ne letter art
Fertilized sl"«&
1978 1979 197B
0.03 (7) d O.O'I 11) cd 0.17 (5) cd
O.ll (6) cd 0.12 CO cd O.'IS (5) b
0.01 («) d 0.06 (5) cd 0.07 (") cd
0.05 ('0 cd 0.07 (") cd 0.23 (") <**
O.l«l (3) e O.'lt (3) de O.Bl (3) cde
0.59 (3) cde 1.30 (3) bcde 2.27 (3) b
0.3H (2) de 0.33 (3) e 0.39 (2) de
0.12 (2) e 0.55 (2) cde I. 10 (2) bcde
1979
0.27 C') >«
1.00 (") "
O.'l'l (M) h
O.'IO C1) b
l.BO (3) bed
6.57 (3) n
1.93 (3) be
2.2H (3) b
of samples is in parentheses; means with in
2 not significantly different, between diftereni
L_ — 4. «^4-~«^,+. fi^"\Ar- / n % n OR niinran'c miiltin
range test).
-------
were not significantly different until after 18 September, when the sludge
plots were slightly lower than the fertilized and controls. Since there were
no distinct differences in the plant communities between treatments this may
reflect a negative sludge effect. However it is also very possible that this
is due to slight differences in plant species composition, or to chance.
In summary, vole populations in the wheat fields showed no apparent
negative response to sewage sludge application. Populations in the sludge
plots were higher than controls due to the fertilizing effect of the sludge.
Populations in the fertilized plots were low due to sparse ground cover re-
sulting from ragweed dominance. Vole populations in the old-fields responded
similarly in all treatments, except for a slight decline in the sludge popu-
lations in late summer. In comparing between communities these results reflect
the greater stability of the old-fields and the ability of the perennial
grasses to resist domination by opportunistic annuals. Research is continuing
along several lines in an attempt to more completely characterize the effect of
sewage sludge application on mammal populations.
190
-------
SECTION 12
POTENTIAL HEALTH EFFECTS FROM VIABLE AND NONVIABLE EMISSIONS
ASSOCIATED WITH SLUDGE INCINERATION AND PYROLYSIS
INTRODUCTION
Combustion is a rapid exothermic oxidation of combustible substances.
J^ls a simple and a quick way of reducing the bulk of human wastes. The
Principal methods for thermal oxidation or degradation of wastewater sludge
now in use or under serious development are incineration (conventional
multiple-hearth furnace or fluidized bed), wet oxidation and pyrolysis. In
some circumstances, sludge may be combined with municipal refuse, for coin-
cineration or co-pyrolysis. The principal characteristics of these methods
are summarized in Table 87 (137,138). Combustion of sludge is a two-step
Process involving drying followed by burning. It is usually accomplished by
Teeding waste into an incandescent chamber (furnace). However, it is usually
aifficult to burn sludge because of its higher moisture content and the
tendency of the wet sludge to resist mixing with combustion air. The primary
Products of combustion are water, CO? and ash. The advantages of combustion
°r thermal degradation of sludge include:
a) nearly complete combustion of organics
t>) large reduction of sludge volumes
0 relatively easy disposal of an inert ash, and
d) the destruction of pathogenic microorganisms.
However, if not properly controlled, thermal degradation can cause air pol-
lution because of smoke, odor and dust emissions.
Incineration is a thermal degradation of a substance in the presence of
oxygen at high temperature and has been used by many cities for a number of
years. Drying and combustion may be accomplished in separate units or suc-
cessively in the same unit, depending upon temperature constraints and control
parameters. Incineration is complete combustion. It is a complex process
involving thermal and chemical reactions which occur at varying times, tem-
peratures, and locations in the furnace. Multiple hearth furnaces are the
roost widely used sludge incinerators in the United States.
Pyrolysis is a destructive distillation of sludge in the absence of ox-
ygen at temperatures somewhat lower than those needed for incineration. In
the pyrolysis process, the volatile matter will distill leaving carbon and
inert residue behind.
191
-------
TABUE 87. CHARACTERISTICS AND POTENTIAL IMPACTS OF THERMAL REDUCTION METHODS (137,138)
Process
Theory and operation
Comments
Potential impacts
VD
ro
Multiple
hearth
incineration
Flash drying
incineration
Fluidized
bed
incineration
Wet oxidation
Counter-current downward flow,
by gravity through hot combus:
tion gases from oil or gas
burners. Rabble arms gently
agitate thin sludge layers
on each successive hearth
to promote drying, then
burning.
Spraying of sludge into large
drying chamber enables sub-
sequent choice of either
burning or utilization of
dried sludge as low-grade
fertilizer.
Upward flow of combustion air
through incandescent bed of
inert particles gives ex-
cellent mixing, good
particle-gas contact.
Use of elevated pressure
and temperature in closed
reactor tank to promote
oxidation of sludge.
Consumes scarce oil or gas,
Inherently entrains fine
flyash in upward-flowing
flue gases. Counter-
current principle very
effective for drying
wet sludge prior to
ignition
Justified only where
markets for low-grade
fertilizer seem
promising.
Inherently entrains
fine flyash in upward-
flowing flue gases.
Reduces volatilization
of metals and organics
in sludge. Operation
requires uncommon skill.
Fine flyash emission which
should be controlled by
available dust collection.
Same as above.
Same as above.
Minor.
(continued)
-------
TABLE 87 (continued)
Process
Theory and operation
Comments
Potential impacts
Combined
refuse/sludge
Pyrolysis
Mixing of sludge with munic-;
ipal refuse and then burning
mixture on travelling or
moving grate.
Thermal decomposition of
carbonaceous and organic
material in a deficiency
of air.
Same as incineration above.
Many full scale attempts
have failed primarily from
problems of nonuniform
burning because of difficulty
of mixing the two dissimilar
components.
Until demonstrated success- Reduce flyash emission less
fully on a full scale plant, volatilization of metals;
the viability of this process smoke and organic emis-
for sewage sludge remains sions have caused some
uncertain. problems.
Reprinted from "Multimedium Management of Municipal Sludge," by National Academy of Sciences,
National Research Council, Analytical Studies for the U.S. Environmental Protection Agency, Volume
IX, 1978, with permission of National Academy of Sciences (l38)«
-------
The potential health effects to sludge disposal workers and the general
population from exposure to pathogens, gaseous and particulate emissions and
ash from incineration and pyrolysis, are considered in this section.
INCINERATION
Incineration converts the organic fraction of the sludge into primarily
C02 and H20, and the inorganic fraction into vapors and ash. The ashes
(mostly containing heavy metal oxides) are usually disposed by landfilling.
Approximately 35% of total sewage sludge is currently incinerated at over
100 facilities in the United States. There are several types of sludge
incinerators commercially available. They are multiple health furnace,
fluidized-bed furnace, electric furnace and single hearth cyclone furnace.
In all furnaces, the drying and combustion processes follow the same phases
- raising the temperature of the feed sludge to 212°F (100°C), evaporating
water from the sludge, increasing the temperature of the water vapor and
air, and increasing the temperature of the dried sludge volatiles to the
ignition point. They differ primarily in the internal arrangement of the
incinerator for the reception and exposure of the material to be burned.
The multiple-hearth furnace is the most popular type of sludge incinerator
in the United States. In this system, sludge is passed through three zones
or hearths. Incineration temperatures range from 800° to 1000°F in the top
hearth where the sludge is dried; 1,400 to 1,800°F in the middle hearth
where volatile gases and solids are burned; and 500 to 600°F in the bottom
hearth. A small proportion of sludge incinerators are fluidized-bed incin-
erators in which sludge is introduced into a bed of sand which has been
preheated to approximately 1,300 to 1,500°F. Combustion in a multiple-hearth
incinerator can destroy nearly 100% of the organics. Disadvantages gen-
erally inv.olve capital cost, ash, odor nuisance and possible explosions.
Sludge parameters that are most important to incineration are moisture,
volatile materials, inert materials and thermal value (139-141). Of these,
moisture is particularly important because of the thermal load it imposes on
the incineration process. Moisture content of sludge is usually reduced by
mechanical dewatering techniques prior to incineration. Volatile and inert
materials affect the heat value of the sludge. Almost all of the com-
bustible substances in sludge are volatile.
Incineration has the advantage of destroying the organic matter present
in sludge, leaving only an odorless sterile ash, as well as reducing the
solid mass by about 90%. Disadvantages include the ash disposal problem and
the potential for air pollution. During the wastewater treatment, the heavy
metals are concentrated in the sludge unless removed prior to incineration.
The metals are not destroyed by incineration but are redispersed into the
atmosphere and also concentrated in the ash. When the ash is landfilled,
these metals may get into groundwater through leaching.
Trace metal composition of sewage sludges from 10 cities in the United
States is shown in Table 88 (142). As can be seen, the concentration of
heavy metals in sewage sludge varies widely from treatment plant to treatment
plant. The concentration also varies with time.
194
-------
TABLE 88. ANALYSIS OF TRACE ELEMENTS IN MUNICIPAL SLUDGES FROM 70 U.S. CITIES
(pprn, dry weight basisj
Element
Arsenic
Barium
Cadmium
Chromium
Cobalt
Copper
Mercury
Manganese
Molybdenum
Nickel
Lead
Zinc
Atlanta
3.6
535
104.2
1320
9.4
1463
6.9
267
6.0
169
1445
2838
Chicago
29
623
14.8
207
3.7
578
6.1
95
10.8
51.2
605
1160
Houston
20.4
485
111.8
3480
6.3
1560
3.8
102
9.8
102
2236
2560
Los
Angeles
26
1066
171.4
4925
16.8
2890
7.1
116
40
402
3065
4590
Miami
10
938
149.6
1430
17.6
1200
15.5
32
37
453
1467
1400
Milwaukee
3.3
344
443,7
14,000
4.3
1288
3.4
134
32.9
360
2253
1370
New
York
8.0
417
29.7
646
6.2
1890
15
152
5.0
140
1976
1340
Philadelphia
16.2
533
192.4
2320
15.6
2680
4.7
95
8.1
432
7627
6890
San
Francisco
6.4
542
8.5
1500
5.3
900
18
113
2.2
223
2521
601
Seattle
30
504
64
1320
9.4
1170
8.2
350
2.3
153
2411
1830
aAdapted from A. K. Furr et
Sewage Sludges of American
al. Multielement and Chlorinated Hydrocarbon Analysis of Municipal
Cities. Env. Sci. and Tech. 10(7):683-687, 1976 (142).
-------
Incineration poses two potential sources of risks to public health - 1)
air pollution from increased ambient air concentration of gaseous and par-
ticulate matter due to emissions from sludge combustion, and 2) possible
groundwater contamination 'from heavy metal leachates from landfilled ash.
The rate of emissions from an incinerator varies widely depending on the
quantity and composition of the sludge, type of the incinerator and the
efficiency of the air pollution control equipment. The degree of public
health hazard posed by emissions from an incinerator will be site-specific
and will depend on the nature of emissions, local meteorological conditions,
ambient air quality levels, etc.
Air pollution is a major concern in any combustion process and may be
the most difficult and expensive environmental consideration to correct.
Incineration might contribute to air pollution because of incomplete com-
bustion and formation of intermediate combustion products (143). The U.S.
Environmental Protection Agency (EPA) has established standards of per-
formance for municipal solid waste incinerators and wastewater sludge incin-
erators (139). These guidelines indicate that new sludge furnaces will have
to comply with the air pollution standards.
Air Pollution Standards
National Ambient Air Quality Standards (NAAQS)-State Implementation Plans
Federal air quality regulations are derived from the Clean Air Act
Amendments of 1970, the Energy Supply and Environmental Coordination Act of
1974, and most recently, the Clean Air Act Amendments of 1977. The NAAQS
established threshold levels of air pollutants below which no adverse effects
would occur. These levels were designed to provide an adequate margin of
safety so as to protect the public health.
Air pollutants are classified into two groups: primary pollutants and
secondary pollutants. Primary pollutants are those emitted directly from
sources, while secondary pollutants are formed by chemical and photochemical
reactions of primary pollutants with the atmosphere. Primary pollutants in-
clude carbon monoxide (CO), hydrocarbons, oxides of nitrogen (NOX), sulfur
dioxide (S02), total suspended particulates (TSP) and lead (Pb). Photo-
chemical oxidants and nitrogen dioxide (N02) are the principal secondary
pollutants. The quantity of secondary pollutants is dependent on the avail-
ability of sunlight as much as on the availability of primary pollutants.
The 1970 Amendments to the Clean Air Act required each state to develop
its own State Implementation Plans (SIP) to meet the federal standards by
1975 or 1977, the date dependent on the severity of the state air quality
problems. For areas already meeting NAAQS standards, implementation plans
must include a program to prevent significant deterioration of air quality.
National Emission Standards for Hazardous Air Pollutants (NESHAPS)--
Subpdrt A of NESHAPS (40 Code of Federal Regulations (CFR)) (63) com-
prises general provisions covering definitions, applications, reporting, and
196
-------
waivers. Subpart E deals with mercury emissions and applies to all operations
that burn or dry wastewater sludge. The NESHAPS standard is currently seven
pounds of mercury (3.2 kg) per 24-hour period for any source (144).
Standards of Performance for New Stationary Sources (NSPS)--
Subpart A of NSPS (40 CFR 60} involves general provisions covering
definitions, performance tests, authority, and monitoring requirements.
Subpart 0 is applicable to incinerators that burn municipal wastewater sludge
and requires that particulates discharged cannot be in excess of 1.30 pounds
per ton (0.65 kg/t) of dry sludge feed and that the gas discharged shall not
have more than 20 percent opacity (145).
New Source Review Standards (NSR)--
This regulation, 40 CFR 51.18, requires a preconstruction review of all
new or modified stationary sources to determine if the source will meet all
applicable emission requirements of the State Implementation Plans and the
USEPA's Emission Offset Policy.
State and local authorities may mandate a stricter criterion. In ad-
dition, the lowest achievable emission rate is required for any regulated
source that mandates Best Available Control Technology (BACT).
Prevention of Significant Deterioration (PSD)--
This regulation, 40 CFR 52.21, limits increases in particulate and
sulfur dioxide concentrations to specified increments above base levels
measured in attainment areas. Data on total emissions for the entire air
basin are required in order to evaluate incremental increases in specific
emissions due to operation of any new or modified furnaces. If the po-
tential emission rate of a regulated pollutant(s) exceeds 250 tons per year
and the allowable emission rate exceeds 50 tons per year then this regu-
lation must be used and public notice is required.
Emi s s 1 ons From _I^nc_rneratj_on
The mixing of air with sludge at high temperature in the incinerator
allows the noncombustible fraction of sludge to become airborne and to be
carried away in the exhaust gases. Thus sludge incineration emits signifi-
cant quantities of fine-grained fly ash (146). In order to be able to meet
the air quality standards all sludge incinerators must be equipped with
scrubbers. Particulate and gaseous emissions for sludge incinerators with
and without scrubbers are listed in Table 89 (138,147). Emission levels for
sludge incinerators with scrubbers may not necessarily meet air quality
standards because the emissions are a function of the collector efficiency.
Several studies have been conducted in recent years on the emission levels
and rates from a number of sludge incinerators. Some of the findings are
summarized below briefly.
Gaseous Emissions— Gaseous pollutants that could be released into the
atmosphere are SO?, nitrogen oxides (NOX), carbon monoxide (CO) and hydro-
chloric acid (HC1). The sulfur content in municipal content is relatively
low, less than 0.1% on a dry weight basis, hence SOg emission is not expected
to be a serious problem. The temperatures employed in sludge incinerators
197
-------
Pollutant
^articulates
Sulfur dioxide
Carbon monoxide
Nitrogen oxides (as NC>2)
Hydrocarbons
Hydrogen chloride gas
Emissions
Uncontrol
Ib/ton
100
1
Neg
6
1.5
1.5
(per unit of dry solids fired)
led After scrubber
kg/MT Ib/ton kq/MT
50
0.5
Neg
3
0.75
0.75
3
0.8
Neg
5
1
0.3
1.5
0.4
Neg
2.5
0.5
0.15
.-_r. ,..„_„ . i VIM • '<*< i w i Mi>wsd i fui iiutiu^wm^iiw \j i riutiiuifJQI J IUUUC » U V ING L I Ul ICE, I
Academy of Sciences, National Research Council, Analytical Studies for the
U.S. Environmental Protection Agency, Volume IX, 1978, with permission of
National Academy of Sciences (138).
198
-------
too low to form appreciable NOX, no more than 200 ppm. Carbon monoxide
is not considered to be a problem if the incinerator is properly operated.
' ne emission of HC1 depends on the amount of combustible plastics in the
sludge. Average levels of approximately 1.0 ppm - 10.0 ppm have been re-
ported for HC1 (146). There are no federal standards for S02 and NOX
emissions from sludge incinerators at present.
, Toxic Organic Chemicals—Toxic organic chemicals can be released from
sludge incineration because of the presence of pesticides and other persistent
organic compounds in sludge. A number of these compounds may concentrate in
tne sludge at wastewater treatment plants. Gabler and Neyland (148) have
snown that pesticides and other organic chemicals can satisfactorily be
aestroyed in a multiple-hearth incinerator.
In a study of incineration of sludge containing low levels of PCBs (1.2
to 2.5 ppm), PCBs were not found in the ash nor in the inlet or outlet
scrubber water. The study showed that 99% of PCBs were destroyed at 1600°
to 1800°F in 2.0 seconds. Ninety-five percent destruction of PCBs was
achieved in a multiple-hearth furnace with no afterburning at normal exhaust
temperatures of about 700°F. The study also showed that complete destruction
PCBs was possible when oxidized with an exhaust gas temperature of about
(146).
Particulates— Available data indicate that on the average stack gases
°T uncontrolled sludge incinerators contain about 1.0 gr/dscf (grains of
Particulates per standard cubic foot of dry gas) or about 45 Ibs of partic-
uiates per ton of sludge burned (141). The federal standard for discharge
or particulates from new and modified municipal sludge incinerators requires
M™Particu1ates discharged cannot be more than 0.65 g/Kg dry sludge input
U.30 lb/ton dry sludge input) (145). This standard applies only to incin-
erators constructed after March 1974 .when the standard became effective.
^articulate control devices must be able to remove approximately 96 to 97
Percent of particulates in incinerator emissions in order to be able to
comply with the standard (148). Many incinerators constructed before the
standard became effective discharge considerably higher levels of air pol-
lutants. in many cases, present sludge incinerator particulate control
systems do meet local and state particulate standards, which are more
stringent than the federal standards.
Heavy Metals— Heavy metals are concentrated in sludge during wastewater
treatment. They are not removed by incineration. They are, however, converted
'nto the less soluble oxide form. Metals that are of most concern are zinc,
cadmium, lead and mercury. Some metals such as lead, mercury and perhaps
cadmium volatilize at sludge incineration temperatures and therefore can be
emitted in gaseous and particulate forms. Even with efficient air pollution
control devices some volatile metals and particulates will be emitted be-
cause of their extremely small submicron sizes. But the impact of these
emissions on the ambient environment is considered to be relatively small
because their initial quantities in sludge are small (138).
Mercury— Mercury presents special problems during incineration. High
combustion temperatures decompose mercury compounds to volatile mercuric
199
-------
oxide or metallic mercury. The Federal emission standard for mercury as a
hazardous air pollutant resulting from sludge incineration established in
1975 limits mercury emission to a maximum of 3,200 g/24-hr period (0.29
lb/hr) (144). Several studies have been conducted to determine the per-
centage of mercury in sludge that leaves the incinerator in the stack gases
(146, 149,150), which indicate that anywhere from 68 to 98 percent of mercury
in sludge will enter the atmosphere. Using an average emission rate of 1.65
g mercury/ metric ton dry solids (146), Los Angeles/Orange County Metro-
politan Area (LA/OMA) engineers (151) calculated that a facility processing
1,939 metric tons of sludge per day would approach the limit. No existing or
planned incinerators approach this size. Thus it is doubtful that any sewage
sludge incinerator would emit more than the federal limit of 3200 gms in 24
hours (152).
Soldano et al. (153) have measured airborne concentrations' of inorganic
and organic mercury compounds near wastewater treatment facilities in several
cities. Both inorganic and organic mercury compounds were detected in a
wide range of concentrations. Although concentrations in downwind samples
were usually higher than those found in upwind samples, frequently concen-
trations increased as downwind distance from the plants increased. Concen-
trations of inorganic mercury were lower than that of organic mercury. In
Louisville, KY, and in Memphis, TN, the levels of both inorganic and organic
compounds were found to be considerably higher than the U.S. Environmental
Protection Agency upper limit of 1000 ng/nn of air. In addition, upwind
samples often had levels as high or higher than downwind to the plant. The
data presented by Soldano, therefore, suggest a source or sources in addition
to the wastewater treatment plants. The presence of the power plants near
the wastewater treatment plants in some cities and other industrial plants
located in the area are probably the sources of the higher mercury levels
found in the ambient air. The report did not specify whether the plants
surveyed had sludge incinerators but it is known from the list of the plants
that some of them do not have incinerators.
Dewling et al. (157) have conducted field studies to measure the emission
of heavy metals at the fluidized bed sludge incinerator, operated by the
Northwest Bergen Sewer Authority, Waldwick, NJ. Their study showed that
about 98% of the mercury in the feed sludge was emitted to the atmosphere
when combustion temperature, averaged 788°C and only 0.4% was retained in
the incinerator ash. Although most of the mercury contained in the feed
sludge was emitted into the atmosphere, the Northwest Bergen facility was
not found to violate the EPA mercury emission limit.
Lead—From a study of emissions from three sludge incinerators, Farrell
et al. (154) showed that under certain conditions as much as 30% of the lead
in sludge can pass through air pollution control devices as fine particulate
matter. Using a plume dispersion model they calculated that the ground
level concentrations of lead from the incinerators were the same order of
magnitude as the annual average ambient concentrations in the cities where
the plants were located. It should be noted that all three incinerators
under study were installed before federal New Source Performance Standards
went into effect.
200
-------
Whitmore and Durfee (150) showed that less than 15% of the lead appeared
in the flue gases of the multiple-hearth incinerator in Palo Alto, California.
Dividing the total daily lead emission by the population served by the in-
cinerator (250,000), Jacknow (155,156) calculated that the per capita dis-
charge of lead was equivalent to driving an automobile using leaded gasoline
with a lead concentration of 2 g/gal less than 200 feet per day (at an assumed
15 mpg rate).
Cadmium—Farre11 et al. (154) showed that under some conditions, up to
30 percent of cadmium in sludge can escape as fine particulate matter. At
the present time, there is no federal ambient air quality standard for cadmium
and the public health implications of elevated cadmium levels associated
with sludge incineration are not known.
Ashi--Incineration is not an ultimate disposal process. About 10-30% of
the total dry mass of sludge is converted to ash which is usually disposed
in a landfill. Heavy metals which usually concentrate in sludge will also
sometimes be concentrated in the ash. Heavy metals from the ash can leach
into groundwater. The impacts of heavy metals in sludge ash in an actual
disposal situation have not been studied yet.
Studies carried out by Dewling et al. (157) showed that Cd and Pb mostly
remained in the ash (80% and 87%, respectively). They did not find evidence
to suggest significant amounts of Pb or cadmium in the off-gas and par-
ticulates emitted from the incinerator.
Gray et al. (158) analyzed incinerator ash from seven municipalities.
'he ashes consisted mainly of oxides of silicon, aluminum, calcium and iron.
AH had basic pH (10.7 to 12.8) and a solubility of 1 to 2 percent dry weight
of ash when leached with about 20 times their mass in water, indicating a
low leachate potential in neutral or slightly alkaline groundwater.
Havens and Emerson (159) have conducted laboratory studies on the leach-
ability of various trace metals in sludges before and after incineration
using a standard citrate insolubility test to simulate the weak organic acid
environment created by soil microorganisms. Compared to digested sludge,
ash showed increased solubility of cadmium and copper, but decreased sol-
ubility of chromium, nickel and zinc. In both sludge and ash, lead showed
no detectable solubility.
Pathogens—High temperatures used in incineration effectively destroy
all pathogens in sewage sludge, leaving a sterile ash. Some incinerators
use secondary effluent from wastewater treatment plants as scrubber water.
Since gases passing through the scrubber water are at temperatures of 150 to
180°F, the temperature at which most viruses are destroyed, the possibility
exists that a few viruses that may be present in the scrubber water might
survive the 150 to 180°F temperature of stack gases and be transported out
°f the incinerator with particulate emissions. EPA tested stack gases of a
single sewage sludge incinerator using a wet scrubber with chlorinated
secondary effluent and found no viruses (152). However, it should be noted
201
-------
that current methodology is not capable of routinely detecting airborne
viruses even at wastewater treatment plants.
There1 may be a potential health hazard from exposure to pathogens in
sludge to workers involved in the several dewatering processes which may
precede sludge incineration.
Potential Health Effects of Incineration
, In addition to the potential health hazard from exposure to pathogens
in sludge to workers involved in the dewatering operations prior to incin-
eration, another potential health risk from incineration is due to air pol-
lution. As discussed in the previous section, the emission of heavy metals
such as mercury, lead, cadmium, arsenic, etc. is not high enough to signifi-
cantly alter the ambient air quality from sludge incineration (146). Incin-
eration also nearly destroys pesticides and persistent organic chemicals
present in sludge, therefore health risks from organic chemicals due to
sludge incineration would be negligible.
Other possible problems are emissions of oxides of nitrogen, particulates,
and gases. Incinerators are not expected to .be a significant source of
atmospheric NOX. The health impact of particulates and gaseous emissions
should be negligible if incinerators are consistently operated such that the
New Source Performance Standards are met. The,potential health effects of
air pollutants are listed in Table 90 (139,160).
.Control of Air Pollution from Incineration
Procedures for curtailing emissions from sludge incinerators include
reducing incineration temperature,^ as in fluidized bed .purhing, \t6 decrease
volatilization of metallic compounds; arid installation- of equipment to
capture the emissions. To meet the new fly ash emission standards, new in-
cinerators must employ high efficiency collectors such as electrostatic
precipitators, wet scrubbers or fabric filters. Some states such as New
York and California require afterburning, i.e., the addition of fuel to
raise the final exhaust temperature to 1,400 to 1,500°F. This destroys
odor-causing compounds and ensures more complete degradation of hydro-
carbons, pesticides and polychlorinated biphenyls.
The New Source Performance Standards (NSPS) set by the USEPA limit
particulate emissions from new sludge incinerators to no more than 70 mg/Nm3
(0.031 grains/standard cubic foot) and less than 20% opacity (145). Meeting
the particulate performance standards requires a fly ash collector, because
simple water sprays and baffle or settling chambers are not efficient enough
to collect such fine dust. Accordingly, high-energy scrubbers, bag houses,
or electrostatic precipitators are needed to achieve compliance with the
standards.
The EPA Sewage Sludge Incineration Task Force (161) concluded that
existing well-designed and properly operated municipal wastewater sludge
incinerators have the capacity to meet the most stringent particulate emis-
sion control regulation existing in any state or local control agency. The
202
-------
o
CO
TABLE 90. HEALTH EFFECTS OF AIR POLLUTANTS (139.160)
Pollutant levels
Air quality . Health
level effect
Significant
harm
General Health Effects
Precautions
ISP*
(24-hour)
jug/m3
S02.
(24-hour)
jug/m3
CO
(8-hour)
1,000 2,620 57,500
Premature death of ill and elderly
adverse symptoms that affect their
All persons should remain indoors,
°3
(1-hour),
jug/m*
1,200
. Healthy people
normal activity.
keeping windows
N02
(1-hour)
/jg/m3
3,750
will experience
and doors closed.
Emergency Hazardous
General Health Effects
Precautions
Warning
Very un-
healthful
General Health Effects
Precautions
All persons should minimize physical exersion and avoid traffic.
875
2,100
40,000
1,000
3,000
Premature onset of certain diseases in addition to significant
aggravation of symptoms and decreased exercise tolerance in
healthy persons.
Elderly and persons .with existing diseases should stay indoors and
avoid physical exersion. General population should avoid outdoor
activity.
625
1,000
34,000
800
2,260
Significant aggravation of symptoms and decreased exercise tolerance in
persons with heart or lung disease, with widespread symptoms in the
healthy population.
Elderly and persons with existing heart or lung disease should stay
indoors and reduce physical activity.
(continued)
-------
TABLE 90 (continued)
ro
o
S02 CO 03 N02
Air quality Health (24-hour), (24-hour), (8-hour), (1-hour), (1-hour),
level effect jug/m3 jug/rrH _/jg/mJ /Jg/mJ ./jg/m-3
Alert Unhealthful 375 800~~17,000 400^1,130
General Health Effects Mild aggravation of susceptible persons, with irritation symptoms
in the health population.
Precautions Persons with existing heart or respiratory ailments should reduce
physical exersion and outdoor activity.
NAAQSC
50 percent
of NAAQSC
Moderate
Good
.260
75^
365
80e
10,000
5,000
240
120
d
d
aTotal suspended particulates.
b400/jg/m3 was used instead of 03 Alert Level of 200/ig/m3.
cNational Ambient Air Quality Standards.
dNo index values reported at concentration levels below those specified by "Alert Level" criteria,
eAnnual primary NAAQS.
-------
newly promulgated federal New Source Performance Standards are based on
demonstrated performance on an operating facility; thus, proper emission
controls and proper operation of the incineration system must be used to
meet all existing regulations of particulate matter.
Residue Disposal
The residues remaining after sludge combustion (ash, particulates from
dry scrubbing, etc.) are typically placed in landfills. The ash from the
combustion of municipal wastewater solids is free of pesticides, viruses and
pathogens. If wastewater effluent is used as scrubber water, it may still
contain low levels of viruses after use. The ash, however, generally contains
high concentrations of trace metals in oxide form. Most of the heavy metal
oxides are less soluble in water except for chromium compounds. Chromium
will be converted to. Cr+6 compounds in the incineration process which are
soluble in water and may cause groundwater pollution from leaching of land-
fill ash. Leachate from sites where incinerator ash is landfilled should be
controlled to prevent contamination of groundwater by metals. Many states
are beginning to classify disposal sites according to their relationship to
nearby groundwater and the material to be landfilled. Tables 91 and 92
describe methods used by the State of California for classifying waste
materials and disposal sites (162). Typically, wastewater sludge incinerator
ash requires a "protected" Class II-l site and municipal refuse incinerator
ash requires a hazardous fill site (139). These are described in Table 92.
Since the sludge ash concentrates the trace metals present in municipal
sludge, recovery of these metals from the ash may be cost effective in
certain special cases. A nationwide study of the concentrations of valuable
metals in..municipal sludges showed that the sludge from Palo Alto, California,
contained some of the highest concentrations of gold, silver, copper and
Platinum of all the locations studied. The treatment plant in Palo Alto,
receives the wastewater from many electronics firms. A private contractor,
through a patented smelting process, has been reported to have extracted
about a half-ounce of gold and 15-20 ounces of silver from each ton of
sludge ash (163).
PYROLYSIS
Pyrolysis is a destructive distillation of sludge in the absence of
oxygen at temperatures of about 800°F which are somewhat lower than those
used in incineration (approximately 1400°F). In the pyrolysis process the
organic portions of the sludge are converted into lower molecular weight
compounds. The products of pyrolysis processes are combustible gases, tar
and oil, and solid char. The gases generated in the pyrolysis process can
be combusted for steam generation thus making it possible to burn sludge.and
other solid wastes with less excess air compared to incineration. This also
results in lower gas volumes for cleaning and higher thermal efficiency.
Classical pyrolysis process is the destructive distillation, reduction
or thermal cracking and condensation of organic matter under heat and/or
Pressure in the absence of oxygen. Complete absence of oxygen requires a
205
-------
TABLE 91. SOLID AND LIQUID WASTE CLASSIFICATION (139,162)
Group 1
Group 2
Group 3
ro
O
CTt
Consist of or contain toxic sub-
stances and substances which
could significantly impair the
quality of usable waters.
Examples include:
• Saline fluids from water
or waste treatment pro-
cesses
. Community incinerator
ashes
• Toxic chemical toilet
waste
• Industrial brines
. Toxic and hazardous fluids
. Pesticides or chemical
fertilizers or their dis-
carded containers
. Other toxic wastes.
Consist of or contain chemically
or biologically decomposable
material which does not include
toxic substances nor those
capable of significantly im-
pairing the quality of usable
waters.
Examples include:
. Garbage
• Rubbish
. Construction debris such as
paper, cardboard, rubber,
etc.
. Refuse such as yard, clippings,
litter, glass, etc.
. Dead animals
. Abandoned vehicles
. Sewage treatment residue such
as solids from screenings and
grit chambers, dewatered sludge
and septic tank pumpings.
. Infectious materials from
hospitals or laboratories.
Consist entirely of nonwater
soluble, nondecomposable inert
solids.
Examples include:
• Construction and
demolition debris,
asphalt paving, inert
plastics, etc.
. Vehicle tires
. Industrial wastes such as
clay products, glass,
slags, tailings, etc.
-------
TABLE 92. CLASSIFICATION OF WASTE DISPOSAL SITES (139,162)
Class I
Class II
Class III
Class III disposal
sites are those at
which protection is
provided to water
quality from Group 3
wastes by location, con-
struction, and operation
which prevent erosion of
deposited material.
ro
o
Class I disposal sites are those at
which complete protection is provided
for all time for the quality of
ground and surface waters from all
wastes deposited therein and
against hazard to public health
and wildlife resources. The
following criteria must be met
to qualify a site as Class I:
(a) Geological conditions are
naturally capable of pre-
venting vertical hydraulic
continuity between liquids
and gases emanating from
the waste in the site and
usable surface or groundwaters.
(b) Geological conditions are
naturally capable of preventing
lateral hydraulic continuity
between liquids and gases
emanating from wastes in the
site and 'usable surface or
groundwaters, or the disposal
area has been modified to
achieve such capability.
(c) Underlying geological for-
mations which contain rock
fractures or fissures of
questionable permeability
must be permanently sealed
to provide a competent
barrier to the movement of
liquids or gases from the dis-
posal site to usable waters.
Class II disposal sites are those at
which protection is provided to water
quality from Group 2 and Group 3
wastes. The types of physical features
and the extent of protection of
groundwater quality divides Class II
sites into the two following
categories;
Class II-l sites are those overlying
usable groundwater. Geologic con-
ditions are either naturally capable
of preventing lateral and vertical
hydraulic continuity between liquids
and gases emanating from the waste in
the site and usable surface or
groundwaters, or the disposal area
has been modified to achieve such
capability.
Class II-2 sites are those having
vertical and lateral hydraulic con-
tinuity with usable groundwater
but for which geological and
hydraulic features such as soil
type, 'artificial barriers, depth to
groundwater, and other factors will
assure protection of the quality
of usuable groundwater underneath
or adjacent to the site.
The following criteria must be met
to quality a site as Class II;
(continued)
-------
TABLE 92 (continued)
Class I
~[cTJinundation ot disposal areas shall not
Class II -
~[aTJ Disposal areas shall be protected by
natural or artificial features so as
to assure protection from any washout
and from inundation which could occur
as a result of tides or floods having
a predicted frequency of once in 100
years.
(b) Surface drainage from the tributary areas
shall not contact Group 2 waters in the
site during disposal operations and for
active life of the site.
(c) Gases and leachate emanating from waste in
the site shall not unreasonably affect
groundwater during the active life of
the site.
(d) Subsurface flow into the site and the
depth at which water soluble materials
are placed shall be controlled during
construction and operation of the site
to minimize leachate production and
assure that the Group 2 waste material
will be above the highest and anticipated
elevation of the capillary fringe of the
groundwater. Discharge from the site
shall be subject to waste discharge
requirements.
occur until the site is closed in
accordance with requirements of the
regional board.
(e) Disposal areas shall not be subject
to washout.
(f) Leachate and subsurface flow into the
disposal area shall be contained within
the site unless other disposition is made
in accordance with requirements of the
regional board.
(g) Sites shall not be located over zones of
active faulting or where other forms of
geological change would impair the com-
r^ petence of natural features or artificial
o° barriers which prevent continuity with
usable waters.
(h) Sites made suitable for use by man-made
physical barriers shall not be located
where improper operation or maintenance
of such structures could permit the
waste, leachate, or gases to contact
usable ground or surface water.
(i) Sites which comply with a, b, c, e, f,
g, and h, but would be subject to
inundation by a tide or a flood of
greater than 100-year frequency may
be considered by the regional board
as a limited Class I disposal site.
-------
sealed chamber with external indirect firing and special construction
materials which make the process expensive and inefficient. Partial pyrolysis
or starved-air combustion is incomplete combustion and occurs when sufficient
oxygen is provided to satisfy the combustion requirements. More conventional
equipment can be used in this process. A number of different reactor types
can be used for pyrolysis. There are two basic types, direct fired and
indirect fired. In an indirect fired reactor, the feed stock materials are
contained in an enclosed vessel and do not come in direct contact with the
heating gases. The retort furnace and indirect fired rotary kiln are ex-
amples of this type of reactor. In a direct fired reactor a fraction of the
stoichiometric oxygen required by the waste is admitted directly into the
fuel fed to liberate the heat'required for pyrolysis or gasification. Al-
ternatively, hot combustion gases may be admitted to the reactor to heat the
feed stock materials. Almost any incinerator reactor can be operated in a
direct fired pyrolysis mode by simply reducing the air supply to less than
the stoichiometric requirement. A multiple hearth furnace is an example of
a reactor which can be operated in a combustion or direct fired pyrolytic
mode. Most of the research and development in the field of pyrolysis of
waste has been directed toward processing of municipal solid waste or
copyrolysis of municipal solid waste and municipal sewage sludge.
At present there are very few pyrolysis projects under development in
the United States that use municipal sludge alone. Several large scale
projects have been proposed and some are in early operation, most work to
1979 has been on laboratory scale (139). Consequently, there is not very
much information available on health impacts of pyrolysis of municipal sludge.
Starved-air combustion, a partial pyrolysis process, had had a number of
successful tests, such as those conducted at the Central Contra Costa
Sanitary 'District (164,165), and several modular combustion units have used
municipal solid waste, sewage sludge, and/or agricultural wastes (166).
Copyrolysis of sewage sludge and municipal solid waste in multiple
hearth furnaces was recommended to the Central Contra Costa Sanitary District
in California, by Brown and Caldwell (164,165). Brown and Caldwell Con-
sulting Engineers conducted pilot tests in a multiple hearth furnace located
in Concord, California, which was once used for sewage sludge incineration
and had been shut down because of high energy costs. The test studies showed
that pyrolysis was easier to control than incineration. The Concord project
also used an afterburner at temperatures in excess of 1400°F'with excess
Pyrolysis gas. This would result in almost complete combustion of organic
chemicals.
The Orange County Sanitation Districts, California, have operated a
Pilot pyrolysis plant using the Activated Carbon Treatment System (ACTS)
developed by the Jet Propulsion Laboratory, Pasadena, California. In this
Process, dewatered raw sludge was converted into a form of activated carbon.
Copyrolysis of municipal refuse and dewatered sewage sludge was carried
°ut in a test program using a PUROX system developed by Union Carbide Cor-
poration in South Charleston, West Virginia (167). The PUROX system consists
of an oxygen-fed shaft furnace in which the solid waste is dried, pyrolyzed,
combusted, and the inorganic residues slagged.
209
-------
Arlington County Water Pollution Control Plant, Virginia, is considering
the use of a multiple hearth incinerator/pyrolyzer combination unit for the
disposal of sewage sludge (168). The pyrolysis system is scheduled to be
completed in 1980.
Starved-air combustion has also had some failures such as the demon-
stration plant in Baltimore, Maryland, which used only municipal solid waste
(139,169). Baltimore used the Monsanto-Landgard process. It was a direct
fired rotary kiln reactor and was used to pyrolyze shredded municipal refuse.
The furnace in Baltimore, Maryland, is now being modified for further testing
and use (139).
Air Pollution from Pyrolysis
The fact that pyrolysis operates without excess air and produces smaller
volume of gas for cleaning than does incineration would indicate that the
emissions from pyrolysis would be much lower than that from incineration
(170). The behavior of the harmful substances in the pyrolysis process in a
pilot experiment were analyzed by Majima et al. (170). Their study showed
that results comparable to that of incineration regarding weight reduction
of sludge can be achieved when pyrolysis is allowed to proceed for an
adequate time period. But they also found that it takes a relatively long
period of time to terminate the gasification reaction at approximately 800°F
because of its low reaction rate.
Since the temperatures used in pyrolysis process are lower than those
needed in incineration, the pyrolysis process may lead to incomplete com-
bustion, leading to gaseous emissions, dust and polycyclic aromatic hydro-
carbons. ..The pyrolysis process may fail to oxidise completely and thus
destroy organics and pesticides. Afterburning, a form of fuel-supported
gaseous incineration, would be required to assure thorough destruction of
such substances.
Heavy Metal s—
The vaporization of the heavy metals would be less than that in in-
cineration, most metals would be concentrated in the char, and as such would
not be of concern in emissions from pyrolysis process (171,172). The trace
metal levels found in the particulates remaining in the product gas after
cleaning with an electrostatic precipitator, in South Charleston, WVA, are
shown in Table 93 (167). These levels are compared with time-weighted average
(TWA) threshold limit values (TLVs) proposed by the American Conference of
Governmental Industrial Hygienists (38). As can be seen from Table 93, heavy
metal emissions from refuse only or from refuse and sludge operation were
far lower than the threshold limit values (TLVs) established for occupational
exposure. Mercury may volatilize at the temperatures used in pyrolysis and,
therefore, should be removed by scrubbers.
In the pyrolysis process, Cr+3 will not be ozidized to Cr+6 since there
is very little air or 02. Cr+3 compounds are insoluble in water and, there-
fore, will not cause groundwater pollution from leaching of landfilled ash
(173).
210
-------
TABLE 93. TRACE METAL CONCENTRATIONS IN PRODUCT GAS
COMBUSTION PRODUCTS* (167)
Metal
Cd
Cr
Cu
Fe
Hg
Pb
Zn
Refuse*5
0.0022
0.0090
0.079
1.62
0.0084
0.038
0.047
Refuse + Sludgec»d
0.0001
0.0007
0.0097
0.03
below detection limit
0.002
0.009
TLV-TWA
0.05
0.5
0.2
5
0.05
0.15
5
Computed from metal content in isokinetically collected particulate in
the product gas.
Refuse-only with particulate recycle and non-optimum gas cleaning.
cBased on average of measurements using primary and mixed sludges with
more optimum gas cleaning performance.
Approximately 5% sludge-dry-solids/refuse.
211
-------
Participates-- . , . . ,
The dust particles generated in the pyrolysis process, in general, are
larger than those from incineration and, therefore, can be removed more
efficiently.
Toxic Organic Chemicals and Pesticides—
The lower temperatures employed in pyrolysis process compared to in-
cineration would lead to incomplete combustion of organic chemicals and may
cause air pollution. Appropriate control measures should be employed for
the control of such emissions.
Pathogens--
Complete destruction is possible at the temperatures employed in
pyrolysis process.
CONCLUSIONS AND RECOMMENDATIONS
Incineration is one of the sludge disposal methods which is capable of
completely destroying pathogens and persistent organic chemicals. Incin-
eration, however, can lead to air pollution because elements such as sulfur,
nitrogen and some volatile metals like mercury and arsenic are converted to
gaseous compounds.
The degree of public health hazard caused by the air pollutants gen-
erated by sludge incinerators will be site specific and will depend on the
nature of emissions, local meteorological conditions, ambient air quality
levels, emissions from other sources, and geography of the region. The
amounts of sulfur and nitrogen oxides produced by sludge incinerators, how-
ever, are'usually small and therefore will only have a minor impact on am-
bient air quality. Also, the air pollution can be reduced or eliminated by
using appropriate control measures. It is therefore concluded that sludge
can be successfully incinerated if the incinerators are properly operated.
At present there are very few pyrolysis projects under development that
use municipal sludge alone in the United States. Several large scale projects
have been proposed and some are in early operation, most work to 1979 has
been on laboratory scale. Consequently, there is not very much information
on health impacts of pyrolysis. However, since pyrolysis process uses lower
temperatures than incineration and since pyrolysis process operates without
excess air, the airborne emission problem from pyrolysis is believed to be
generally less difficult to manage than that from incineration. This would
be true of heavy metals but the organic chemicals in sludge may only undergo
incomplete combustion at the temperatures used in the pyrolysis process
producing emissions requiring appropriate control measures for successful
operation. After burning, a form of fuel-supported gaseous incineration,
if used, can assure complete combustion of such substances.
212
-------
REFERENCES
1. Clark, C. S., E, J. Cleary, G. M. Schiff, C. C. Linnemann, Jr., J. P.
Phair and T. M. Briggs. Disease Risks of Occupational Exposure to
Sewage. J. Environ. Eng. Div. Am. Soc. Civil Engineers, 102(EE2):
375-388, 1976.
2. Clark, C. S. et al. Health Risks of Human Exposure to Wastewater.
Final Report, U.S. Environmental Protection Agency, Health Effects
Research Laboratory, Grant No. R 803643, Cincinnati, Ohio, 1980.
3. Mi liner, P. D., P. B. Marsh, R. B. Snowden and J. F. Parr. Occurrence
of Aspergillus fumigatus During Composting of Sewage Sludge. Appl.
Environ. Microbiol. 34(6):767-772, 1977.
4. Boswell, F. C. Municipal Sewage Sludge and Selected Element Appli-
cations to Soil: Effect on Soil and Fescue. J. Environ. Qual. 4:
267-272, 1975.
5. Furr, A. K., G. S. Stoewsand, C. A. Bache and D. J. Lisk. Study of
Guinea Pigs Fed Swiss Chard Grown on Municipal Sludge Amended Soil:
Multi-element Content of Tissues. Arch. Environ. Health 31:87-91,
1976.
6. Hinesly, T. D., R. L. Jones, J. J. Tyler and E. L. Ziegler. Soybean
Yield Responses and Assimilation of Zn and Cd from Sewage Sludge
Amended Soil. J. Water Poll. Con. Fed. 48:2137-2152, 1976.
7. Kinzell, J. H., P. R. Cheeke and R. W. Chen. Tissue Heavy Metal
Accumulation, Pentobarbital Sleeping Times and Multigeneration Re-
productive Performance of Rats Fed Activated Sewage Sludge. Nutr.
Rep. Intern. 15:645-650, 1977.
8. Process Design Manual. Sludge Treatment and Disposal. Municipal
Environmental Research Laboratry, U.S. Environmental Protection
Agency Report, EPA-600/1-79-011, 1979.
9. Environmental Resources Company. Workshop on the Health and Legal
Implications of Sewage Sludge Composting, Massachusetts, December
18-20, 1978.
10. Detroy, R. W. et al. Aflatoxin and Related Compounds. In: Microbial
Toxins: A Comprehensive Treatise, Vol. VI, Fungal Toxins, A. Ciegler,
S. Kadis and S. J. Ajl, Eds., New York, Academic Press, Inc., 1971.
213
-------
11,. Rylander, R., K. Andersson, L. Bel in, G. Berglund, R. Bergstrom, L.
Hanson, M. Lundholm and I. Mattsby. Studies on Humans Exposed to
Airborne Sewage.,Sludge. Schweir. Med. Wschr. 107:182-184, 1977.
12. Mattsby, I. and R. Rylander. Clinical and ImmunoTogical Findings in
Workers Exposed to Sewage Oust. Jour, of Occup. Med. 20(10):690-692,
1978.
13. Rylander, R., K. Andersson, C. Belin, G. Berglund, R. Bergstrom, C.
Hanson, M. Lundholm and I. Mattsby. Sewage Workers Syndrome. Lancet
28:478-479, 1976.
14. Rylander, R. et al. Local and Systemic Immune Response to Cotton
Dust Bacteria. Paper presented at the 3rd Special Session on Cotton
Dust, The Beltwide Cotton Conference, Phoenix, Arizona, 1979.
15. Rylander, R. Exposure to Gram-Negative Bacteria and the Development
of Byssinosis. Paper presented at the XIX International Congress
on Occupational Health, Dubrounik, Yugoslavia, 1978.
16. McGuire, W. et al. Activation of Plasma Contact System in In-
flammatory Lung Disease. Federation Proceedings 39:906, 1980.
17. Dutkiewiez, J. Exposure to Dust-Bome Bacteria in Agriculture. I.
Environmental Studies. Arch. Environ. Health 33(5}:250-259, 1978.
18. Dutkiewiez, J. Exposure to Dust-Borne Bacteria in Agriculture. II.
Immunological Survey. Arch. Environ. Health 33(5):260-270, 1978.
19. Lundholm, B. and R. Rylander. Occupational Symptoms Among Compost
Workers. J. Occup. Med. 22(4):256-257, 1980.
20. Millner, P. D., D. A. Bassett and P. B. Marsh. Dispersal of
Aspergillus fumigatus from Sewage Sludge Compost Piles Subjected to
Mechanical Agitation in Open Air. Appl. Environ. Microbiol. 39(5):
1000-1009, 1980.
21. May, K. R. Calibration of a Modified Andersen Bacterial Aerosol
Sampler. Appl. Microbiol. 12(l):37-43, 1964.
22. Passman, F. J. Energy Resources Co. Personal Communication, November
12, 1979.
23. Clark, C. S., H. S. Bjornson, J. W. Holland, T. L. Huge, V. A. Majeti
and P. S. Gartside. Occupational Hazards Associated with Sludge
Handling. Chapter 10 in G. Bitton, B. L. Damron, G. T. Edds and J. M.
Davidson, Eds. Sludge-Health Risks of Land Application, Ann Arbor
Science, Ann Arbor, Michigan, 1980.
214
-------
24. Rippon, J. W. Medical Mycology: The Pathogenic Fungi and the
Pathogenic Actinomycetes, W. B. Saunders Co., Philadelphia, Pennsylvania,
1974.
25. Westphal, 0. and K. Jans. Bacterial Lipopolysaccharides. Extraction
with Phenol-Water and Further Applications of the Procedures. In:
Methods in Carbohydrate Chemistry, Vol. 5, R. L. Whistler and M. L.
Wolfrom, Eds., New York, Academic Press, Inc., 1965, pp. 83-91.
26. Gewurz,.H. and L. A. Sujehira. Complement. In: Manual of Clinical
Immunology, N. R. Rose and H. Friedman, Eds., American Society for
Microbiology, Washington, D.C., 1976, p. 36.
27. Engrail, E. and P. Perlmann. Enzyme-Linked Immunosorbent Assay. III.
Quantitation of Specific Antibodies by Enzyme-Labeled Anti-Immunoglobulin
in Antigen Coated Tubes. J. Immunol. 109:129-134, 1972.
28. Coleman, R. M. and L. Kaufman. Use of the Immunodiffusion Test in
the Serodiagnosis of Aspergillus. Appl. Microbiol. 23(2):301-308,
1972.
29. Mancini, G., A. D. Carbonera and J, F. Heremans. Immunochemical
Quantitation of Antigens by Single Radial Immunodiffusion. Immunochem.
2:235-239, 1965.
30. APHA, AWWA, WPCF. Standard Methods for the Examination of Water and
Wastewater. 14th Edition, American Public Health Association,
Washington, D.C., 1976.
31. Conover, W. J. Practical Nonparametric Statistics, Wiley and Sons,
New York, New York, 1971.
32. Rima, D. R., E. Brown, D. F. Goerlitz and L. M. Law. Potential Con-
tamination of the Hydrologic Environment from the Pesticide Waste Dump
in Hardeman County, Tennessee. Administrative Report of the U.S.
Geological Survey to the Federal Water Pollution Control Administration,
August, 1967.
33. Tennessee Stream Pollution Control Board. Transcript of Hearing on
the Velsicol Chemical Company Waste Disposal Site in Hardeman County.
Jackson, Tennessee, March 4, 1971.
34. Sprinkle, C. L. Leachate Migration from a Pesticide Disposal Site in
Hardeman.County, Tennessee. U.S. Geological Survey, Open File Report
78-128, Nashville, Tennessee, 1978.
35, Clark, C. S., V. A. Majeti and V. J. Elia. Urine Screening of Workers
Exposed to Toxic Waste Chemicals in a Municipal Wastewater Treatment
Plant. University of Cincinnati, Department of Environmental Health
Annual Report, Cincinnati, Ohio, 1979, pp. 224-227.
215
-------
36. U.S. Environmental Protection Agency, Region IV. Summary of USEPA and
State of Tennessee Chemical Analysis. Atlanta,' GA, March 9, 1979.
37. Keith, L. H. and W. A. Telliard. Priority Pollutants I - A Perspective
View. Environ. Sci. Technol. 13(4):416-423, 1979.
38. Threshold Limit Values for Chemical Substances and Physical Agents in
the Workroom Environment with Intended Changes for 1977. American
Conference of Governmental Industrial Hygienists, Cincinnati, Ohio,
1979.
39. Elia, V. J., C. S. Clark and V. A. Majeti. Evaluation of Worker Ex-
posure to Organic Chemicals at Municipal Wastewater Treatment
Facilities. University of Cincinnati, Department of Environmental
Health Annual Report, Cincinnati, Ohio, 1979, pp. 228-232.
40. Robbins, B. H. The Absorption, Distribution, Excretion of Carbon
Tetrachloride in Dogs Under Various Conditions. Jour. Pharmacol.
37:203, 1929.
41. Barrett, H. M., J. G. Cunningham and 0. H. Johnson. Study of Fate in
Organisms of Some Chlorinated Hydrocarbons. Jour. Ind. Hyg. Toxicol.
21:479, 1939.
42. U.S. Environmental Protection Agency. Preliminary Assessment of
Suspected Carcinogens in Drinking Water. Office of Toxic Substances,
U.S. EPA, Washington, D.C., 1975.
43. U.S. Environmental Protection Agency. Preliminary Assessment of
Suspected Carcinogens in Drinking Water. Appendices. EPA 560/4-75/
003A. U.S. Environmental Protection Agency, Washington, D.C., 1977.
44. U.S. Environmental Protection Agency. OWPS Criterion Document-on
CC14, 1978.
45. American Chemical Society. Carbon Tetrachloride Spill Causes Stir
Along the Ohio River. Chem. Engg. News 55:7, February 28, 1977.
46. National Research Council. Nonfluorinated Halomethanes in the En-
vironment. National Academy of Sciences, Washington, D.C., 1978.
47. Lillian, D. et al. Atmospheric Fates of Halogenated Compounds.
Environ. Sci. & Technol. 9(12):1042-1048, 1975.
48. National Academy of Sciences. Chloroform, Carbon Tetrachloride and
Other Halomethanes: An Environmental Assessment. Washington,'D.C.,
1978.
49. Louria, D. B. and J. D. Bogden. The Dangers from Limited Exposure to
Carbon Tetrachloride. CRC Critical Reviews in Toxicology 7(2):177-
188, 1980.
216
-------
50. Ingle, L. The Toxicity of Chlordane Vapor. Science 118:213, 1953.
51. Carter, M. R. The Louisville Incident. Internal Report (unpublished).
Surveillance and Analysis Division, Region IV. U.S. Environmental
Protection Agency, Athens, GA, 1977.
52. National Institute for Occupational Safety and Health. Registry of
Toxic Effects of Chemical Substances. 403, 1978.
53. Treon, T. F., (v. P. Cleveland and J. Cappet. The Toxicity of
Hexachlorocyclopentadiene. Indust. Health 11:459-472, 1955.
54. U.S. Environmental Protection Agency. Review of the Environmental
Effects of Pollutants. XI. Hexachlorocyclopentadiene. EPA-600/1-
78-047, Cincinnati, Ohio 1978.
55. U.S. Environmental Protection Agency. Hexachlorocyclopentadiene.
Ambient Water Quality Criteria, Washington, D.C., 1979.
56. Spehar, R. L. et al. Toxicity and Bioaccumulation of Hexachloro-
cyclopentadiene, Hexachloronorbornadiene and Heptachloronorbornene in
Larval and Early Fathead Minnows. Pimephales Promelas, Bull. Environ.
Cont. Toxicol. 21:576-583, 1979.
57. Cody, T. E., V. J. Elia and C. S. Clark. Preliminary Toxicity and
Mutagenicity Evaluation of Groundwater Contaminated by an Abandoned
Landfall. University of Cincinnati, Department of Environmental
Health Annual Report, Cincinnati, Ohio, 1979, pp. 202-207.
58. Simmon, V. F. Affidavit U.S. District Court, Western District of
Tennessee, Western Division, U.S.A. vs. Velsicol Chemical Corporation,
C.ivil Action No. C-78-2335, July 1978.
59. Rima, D. R. Susceptibility of the Memphis Water Supply to Contam-
ination from the Pesticide Waste Disposal Site in Northeastern Hardeman
County, Tennessee. U.S. Geological Survey, Open File Report 79-750,
Nashville, Tennessee, 1979.
60. Mississippi-Arkansas':Tennessee Council of Governments, Memphis Delta
Development District. Section 208 Areawide Waste Treatment Management
Plan, Waste Treatment Facilities Plan, Vol. 1, Chapter 1, History of
Wastewater Collection System Development and Existing Wastewater Dis-
charge Information, March 4, 1977.
61. Environmental Management Planning and Engineering, Inc. Influent Hazard
Analysis, Memphis North Sewage Treatment Plant, Memphis, Tennessee,
August-September 1977.
62. Communication, Augu.st 30, 1977 from Acting Chief, Hazard Evaluation and
Technical Assistance Branch, National Institute of Occupational Safety
and Health to Mayor, City of Memphis, Tennessee.
217
-------
63. Morse, D, L.t J. R. Kominsky, C. L. Wissemann, III and P. J. Landrigan.
Occupational Exposure to Hexachlorocyclopentadiene: How Safe is Sewage?
JAMA 241:2177-2179, 1979. ... •
64. Kominsky, J. R., C. L. Wisemann, III and D. 1. Morse. Hexachloro-
cyclopentadiene Contamination of a Municipal Wastewater Treatment Plant.
Am. Ind. Hyg. Assoc. Jour. 41(8):552-556, 1980.
65. Elia/'V. J., C. S. Clark, V. Majeti, T. Macdonald and N. Richdale.
Worker Exposure to Organic Chemicals at an Activated Sludge Wastewater
Treatment Plant. In: Proceedings of U.S. Environmental Protection
Agency Symposium on Wa-stewater Aerosols and Disease, Cincinnati, Ohio,
September 1979.
66. Majeti, V. A. and C. S. Clark. Health Risks of Qrganics in Land
Application. J. Environ. Eng. Div. Am. Soc. Civil Engineers (accepted
for publication, 1980).
67. Threshold Limit Values for Chemical Substances and Physical Agents in
the Workroom Environment with Intended Changes for 1977. American
Conference of Governmental Industrial Hygienists, Cincinnati, Ohio,
1979.
68. Spehar, R. L. et al. Toxicity of Bioaccumulation of Hexachlorocyclo-
pentadiene, Hexachloronorbornadiene and Heptachloronorobornene in
Larval and Early Juvenile Fathead Minnows. Bull. Env. Cont. Toxicol.
21:576-583, 1979.
69. Thomas, T. C. and J. N. Seiben. Chromosorb 102, An Efficient Medium
for Trapping Pesticides from Air. Bull. Environ. Cont. Toxicol. 12:
17-25, 1974.
70. Neumeister, C. E. and R. W. Kurimo. Determination of Hexachlorocyclo-
pentadiene and Octachlorocyclopentene in Air. Presented at American
Industrial Hygiene Conference, May 7-17, Los Angeles, CA.
71. U.S. Department of Health Education and Welfare: NIOSH Manual of
Analytical Methods, Vol. 5. NIOSH, Cincinnati, Ohio, 1977.
72. Burgessar, A. J. and J. F. Colaruotolo. Extraction of Semi- and Non-
volatile Chlorinated Organic Compounds from Water. Anal. Chem. 51:
1588-1589, 1979.
73. Ingle, L. The Toxicity of Chlordane Vapor. Science 118:213, 1953.
74. Treon, T. F., F. P. Cleveland and J. Cappet. The Toxicity of Hexa-
chlorocyclopentadiene. Indust. Health 11:459-472, 1955.
75. Carter, M. R. The Louisville Incident. Internal Report (unpublished),
Surveillance and Analysis Division, Region IV, U.S. Environmental Pro-
tection Agency, Athens, GA, 1977.
218
-------
76. Mehendale, H. M. The Chemical Reactivity-Absorption Retention,
Metabolism and Elimination of Hexachlorocyclopentadiene. Environ.
Health Perspect. 21:275-278, 1977.
77. Demerjian, Y. A., D. R. Kendrick and T. R. Westman. Muskegon County
Wastewater Management System: Report on Major Research Activities,
1972-1978. Muskegon County Department of Public Works, Muskegon.
78. The Muskegon County Plan of Wastewater Reuse. Public Works, October
1973.
79. Environmental Research Corporation. Biological Sampler/Collector
XM2 Operator's Manual, December 18, 1974, St. Paul, MN.
80. Dahling, D. R. and R. S. Safferman. Survival of Enteric Viruses
Under Natural Conditions in a Subarctic River. Appl. and Environ.
Microbiol. 38(6):1103-1110, 1979.
81. Dulbecco, R. and M. Vogt. Plaque Formation and Isolations of Pure
Lines with Poliomyelitis Viruses. J. Exp. Med. 99:167-182, 1954.
82. Hsiung, G. D. and J. L. Melnick. Plaque Formation with Polio-
myelitis, Coxsackie and Orphan (ECHO) Viruses in Bottled Cultures
of Monkey Epithelial Cells. Virology 1:533-535, 1955.
83. Rao, V. C., U. Chandorkar, N. U. Rao, P. Kumaran and S. B. Lakhe. A
Simple Method for Concentrating and Detecting Viruses in Wastewater.
Water Res. 6:1565-1576, 1972.
84. Safferman, R. S. and M. Morris. Assessment of Virus Removal by a
Multi-Stage Activated Sludge Process. Water Res. 10:413-420, 1976.
85. Adams, M. H. Bacteriophages. Interscience Publishers, New York,
1959.
86. Kott, Y. Estimation of Low Numbers of Escherichia Coli Bacteriophage
by Use of the Most Probable Number Method^Appl. Micro. 14(2):141-
144, 1966.
87. Scarpino, P. V. Bacteriophage Indicators. In: Indicators of
Viruses in Water and Food, 6. Berg, Ed., Ann Arbor Science Publishers,
Inc., Ann Arbor, MI, 1978.
88. Air Sampling Instruments for Evaluation of Atmospheric Contaminants.
4th Edition, American Conference of Governmental Industrial
Hygienists, Cincinnati, Ohio, 1972.
89. Andersen, A. A. New Sampler for the Collection, Sizing and
Enumeration of Viable Airborne Particles. Jour. Bacteriol. 76(5):
471-488, 1958.
219
-------
90. Fair, G. M. and W. F. Wells. Measurement of Atmospheric Pollution and
Contamination by Sewage Treatment Works. Proceedings of 19th Annual
Meeting New Jersey Sew. Works Assn., Trenton, N.J. 20, 1934.
'91. ATbrecht, C. R. Bacterial Air Pollution Associated with the Sewage
Treatment Process. M.S. Thesis, University of Florida, Gainesville,
August 1958.
92. Ladd, F. C. Airborne Bacteria from Liquid Waste Treatment Units. M.S.
Thesis, Oklahoma State University, Stillwater, 1966.
93. Carnow, B., R. Northrop, R. Wadden, S. Rosenberg, J. Holden. A. Neal,
L. Sheaff, P. Scheff and S. Meyer. Health Effects of Aerosols Emitted
from an Activated Sludge Plant. EPA-600/1-79-019, U.S. Environmental
Protection Agency, Cincinnati, Ohio, May 1979.
94. Johnson, D.E., D. E. Camann, H. J. Harding and C. A. Sorber. En-
vironmental Monitoring of a Wastewater Treatment Plant. EPA-600/1-79-
027, U.S. Environmental Protection Agency, Cincinnati, Ohio, August
1979.
95. Johnson, D. E., D. E. Camann, J. W. Register, R. E. Thomas, C. A.
Sorber, M. N. Guentzel, J. M. Taylor and H. J. Harding. The Evaluation
of Microbiological Aerosols Associated with the Application of Waste-
water to Land: Pleasanton, California. EPA-6QO/1-80-015, U.S. En-
vironmental Protection Agency, Cincinnati, Ohio, 1980, 191 pp.
96. Bausum, H. T., S. A. Schaub and C. A. Sorber. Viral and Bacterial
Aerosols at a Wastewater Spray Irrigation Site. NSF/RA-761319, Pre-
sented at 76th Annual Meeting, American Society for Microbiology,
Atlantic City, N.J., May 3, 1976, U.S. Army Medical Bioengineering
Research and Development Laboratory, Fort Detrick, Frederick, MD.
97. Randall, C. W. and J. 0. Ledbetter. Bacterial Air Pollution from
Activated Sludge Units, Amer. Indus. Hyg. Assoc. Jour. 27:(6):506-
519, November-December 1966.
98. Cronholm, L. S. Emission of Microbial Aerosols from Polluted Waters
in Densely Populated Regions. Research Report No. 117, University
of Kentucky, Lexington, KY, U.S. Department of the Interior, 1978.
99. Finch, J.E., J. Prince and M. Hawksworth. A Bacteriological Survey of
the Domestic Environment. Microbiol. Lab. Pharmaceutical Div.,
Reckitt and Colman Ltd, Hull HU8 70S, UK, J. Appl. Bacteriol. 45(3):
357-364, 1978.
100. Bagley, S. T., R. J. Seidler, H. W. Talbot, Jr. and J. E. Morrow.
Isolation of Klebsielleae from Within Living Wood. Appl. Environ.
Microbiol. 36(1):178-185,1978.
220
-------
101. Km'ttel, M.D., R. J. Seidler, C. Eby and L. M. Cabe. Colonization of
the Botanical Environment by Klebsiella Isolates of Pathogenic Origin.
Appl. Environ. Microbiol 34(5):557-563, 1977.
102. Ducluzeau. R. S. Hudault and J. V. Galpin. Longevity of Various
Bacterial Strains of Intestinal Origin in Gas Free Mineral Water.
Eur. J. Appl. Microbiol. 3(3):227-236, 1976.
103. Gevaudan, M.-J., A. Blancard, C. Gulian and M.-N. Mallet. Resistance
of Pseudomonas Aeruginosa and Klebsiella Pneumoniae in the Exterior
Environment. Mediterr. Med. 3(74):13-16, 1975.
104. Varma, M. M., W. A. Thomas and C. Prasad. Resistance to Inorganic
Salts and Antibiotics Among Sewage-Borne Enterobacteriaceae and
Achromobacteriaceae. J. Appl. Bacteriol. 41(2):347-349, 1976.
105, Levi, J. A., S. C. Schimpff, R. G. Slawson and P. H. Wiernik. Evaluation
of Radiotherapy for Localized Inflammatory Skin and Perianal Lesions
in Adult Leukemia: A Prospectively Randomized Double Blind Study.
Cancer Treat. Rep. 61(7):1301-1305, 1977.
106. Umenai, T., Y. Saitoh and M. Chiba. Septicemia Caused by Klebsiella
oxytoca. Tohku J. Exp. Med. 124(4):393-394, 1978.
107. Gross, G. N., S. R. Rehm and A. K. Pierce. .The Effect of Complement
Depletion on Lung Clearance of Bacteria. J. Clin. Invest. 62{2):
373-378, 1978.
108. Harris, G. D., W. G. Johanson, Jr. and A. K. Pierce. Determinants
of Lung Bacterial Clearance in Mice After Acute Hypoxia. Am. Rev.
Respir. Dis. 116(4):67i-677, 1977.
109. Flint, L. M., Jr., C. R. Voyles, J. D. Richardson and D. E. Fry.
Missile Tract Infections After Transcolonic Gunshot Wounds. Arch.
Surg. 113(6):727-728, 1978.
110. Baergaluppi, J. F., R. Negrom and H. M. de Severino. Ann. Allergy
42(2):95-98, 1979.
111. Mackay, D. and A. W. Wolkoff. Rate of Evaporation of Low-Solubility
Contaminants from Water Bodies to Atmosphere. Environ. Sci. Technol.
7:611-614, 1973.
112. Dilling, W. L.} N. B. Teforttiller and C. J. Kallos. Evaporation Rates
and Reactivities of Methylene Chloride, Chloroform 1,1,1-Trichloro-
ethane, Tetrachloroethlene and Other Chlorinated Compounds in Dilute
Aqueous Solutions. Environ. Sci. Technol. 9:833-873, 1975.
113. Thibodeaux, L. J. and J. D. Millican. Quantity and Relative De-
sorption Rates of Air-Strippable Organics in Industrial Wastewater.
Environ. Sci. Technol. 11:879-883, 1977.
221
-------
114. Maynard, J. E., D. W. Bradley, C. L. Hornbeck, R. M. Fields, I. L.
Doto and F. B. Hollinger. Preliminary Serologic Studies of Antibody
to Hepatitis A Virus in Populations in the United States. J. Infect.
Dis. 134:528-530, 1976.
115. Hall, W. T., F. K. Mundon and D. L. Madden. Antibody to Hepatitis A
in Mentally Retarded Inpatients. The Lancet, Arpil 1977, pp. 78-9.
116. Rose, F. L. and J. C. Harshbarger. Neoplastic and Possibly Related
Skin Lesions in Neotenic Tiger Salamanders from a Sewage Lagoon.
Science 196:315-317, 1977.
117. Tipton, I. H. and M. J. Cook. Trace Elements in Human Tissue: Part
II. Adult Subjects for the United States. Health Physics 9:103,
1963.
118. Kinzell, J. H., P. R. Cheeke and R. W. Chen. Tissue Heavy Metal
Accumulation, Pentobarbital Sleeping Times and Multigeneration Re-
productive Performance of Rats Fed Activated Sewage Sludge. Nutr.
Rep. Intern. 15:645-650, 1977.
119. Odum, E. P., G. W. Barrett and H. R. Pulliam. Testing Pesticides at
the Ecosystem Level. ASB Bull. 15:48, 1968.
120. Barrett, G. W. The Effects of an Acute Insecticide Stress on a Semi-
Enclosed Grassland Ecosystem. Ecology 49:1019-1035, 1968.
121, Barrett, G. W., G. M. Van Dyne and E. P. Odum. Stress Ecology. Bio-
science 26:192-194, 1976.
122. Odum, E. P. The Emergence of Ecology as a New Integrative Dis-
cipline. Science 195:1289-1293, 1977.
123. Woodwell, G. M. Recycling Sewage Through Plant Communities. Amer.
Sci. 65:556-562, 1977.
124. Odum, E. P. the Strategy of Ecosystem Development. Science 164:
262-270, 1969.
125. Sopper, W. E. and L. T. Kardos. Recycling Treated Municipal Waste-
water and Sludge Through Forest and Cropland. Penn State University
Press, University Park, PA, 1973, 494 pp.
126. .Odum, E. P. Organic Production and Turnover in Old-Field Succession.
Ecology 41:34-49, 1960.
127. Golley, F. B. Structure and function of an Old-Field Broomsedge
Community. Ecol. Manage. 35:113-131, 1965.
128. Wiegert, R. G. and F. C. Evans. Primary Production and the Dis-
appearance of Dead Vegetation on an Old-Field in Southeastern Michigan.
Ecology 45:49-63, 1964.
222
-------
129. Hurd, L. E., M. V. Mellinger, L. L. Wolf and s. J. McNaughton.
Stability and Diversity at Three Trophic Levels in Terrestrial
Successional Ecosystems. Science 173:1134-1136, 1971.
130. Krebs, C. J., B. L. Keller and r. H. Tamarin. Microtus Population
Biology: Demographic Changes in Fluctuating Population of M.
pchrogaster and M. pennsylvanicus in Southern Indiana. Ecology 50:
587-607, 1969.
131. Pitelka, F. A. and A. M. Schultz. The Nutrient-Recovery Hypothesis
for Arctic Microtine Cycles. In: Population Ecology, L. Adams, Ed.,
Dickerson Pub. Co., Belmont, CA, 1970, 160 pp.
132. Taylor, W. G., G. W. Barrett and T. J. Anderson. The Effects of
Sewage Sludge and Fertilizer on Primary Productivity and Diversity
in an Annual and a Perennial Grassland Community. In review, Amer.
Midi. Nat.
133. Carson, W. P., T. A. Boggs and G. W. Barrett. The Effects of Dried
Sewage Sludge on a 1-Year and a 5-Year Old-Field. In preparation,
J. Environ. Mgmt.
134. Sedlacek, J. D. and G. W. Barrett. Effects of Dried Sewage S'iudge
on Density and Diversity of Anthropod Communities in an Annual and a
Perennial Grassland. In Preparation.
135. Anderson, T. 0., G. W. Barrett, K. L. Stemmer, C. S. Clark, V. J. Elia,
V. A. Majeti and T. E. Cody. Tissue Metal Concentration and
Pathology in Meadow Voles from Sewage Sludge-Amended Fields. In
preparation.
136. Anderson, T. J. and G. W. Barrett. Effects of Dried Sewage Sludge
on Meadow Vole Populations in Two Grassland communities. In review,
J. Appl. Ecol.
137. Burd, R. S. A Study of Sludge Handling and Disposal. Federal Water
Pollution Control Research Service, Publication No. WP-20-4. Wash-
ington, D.C., Federal Water Pollution Control Administration, 1968.
138. National Academy of Sciences. Multimedium Management of Municipal
Sludge. National Research Council. Analytical Studies of the U.S.
Environmental Protection Agency, Volume IX, 1978.
139. U.S. Environmental Protection Agency. Sludge Treatment and Disposal.
EPA-625/1-79-011, Cincinnati, Ohio, 1979. Chapter 11.
140. Russell, R. A. Theory of Combustion of Sludge. Sludge Concentration-
Filtration and Incineration. School of Public Health, University of
Michigan, Continued Education Series No. 113, Ann Arbor, Michigan,
University of Michigan (out of print), 1964, pp. 152-157.
223
-------
141. Shen, T. T. Air Pollution from Sewage Sludge Incineration. Pro-
ceedings of the American Society of Civil Engineers, Jour, of Env. Eng.
Div. 105(EEl):61-74, 1979.
142. Furr, A. K. et al. Multielement and Chlorinated Hydrocarbon Analysis
of Municipal Sewage Sludges of American Cities. Env. Sci. and Techno!.
10(7):683-687, 1976.
143. Water Pollution Control Federation. Design of Wastewater Treatment
Plants.. Washington, D.C., Water Pollution Control Federation, 1977.
144. Federal Register 40(199):48292, October 14, 1975.
145. Devitt, T. W. and N. J. Kulujian. Sewage Sludge Incinerators-In-
spection Manual for the Enforcement of New Source Performance
Standards. EPA-340/1-75-004, U.S. Environmental Protection Agency,
Washington, D.C., February 1975.
146. U.S. Environmental Protection Agency. Air Pollution Aspects of
Sludge Incineration. Office of Technology Transfer, Seminar Publi-
cation, EPA-625/4-75-009, Washington, D.C., 1975.
147. U.S. Environmental Protection Agency. Land Treatment of Municipal
Wastewater Effluents: Design Factor-I. EPA-625/4-76-010, Vol. 1.
January 1976, PB-259955.
148. Gabler, R. C., Jr. and D. L. Neyland. A Study of Pesticide Disposal
in a Sewage Sludge Incinerator. Versar, Inc., Monthly Progress Re-
port, EPA Contract No. 68-01-1587, U.S. Environmental Protection Agency,
Washington, D.C., September 1974, PB-253 485/7BE.
149. U.S. Environmental Protection Agency. Background Information on
National Emission Standards for Hazardous Air Pollutants - Proposed
Amendments for Asbestos and Mercury. EPA-450/2-74-0099, 1974a.
150. Whitmore, F. C. and R. J. Durfee. Lead and Mercury Balance at the
Palo Alto Incinerator. Versar, Inc., 1974.
151. Los Angeles/Orange County Metropolitan Area (LA/OMA). Sludge Pro-
cessing and Disposal - A State-of-the-Art Review. Regional Wastewater
Solids Management Programs, California, 1977.
152. Workshop on the Health and Legal Implications of Sewage Sludge Com-
posting. Vol. 2, Chapter 9, Environmental Resources company, December
18-20, 1978, Cambridge, Massachusetts.
153. Soldano, B. A., P. Bien and P. Kwan. Airborne Organo Mercury and
Elemental Mercury Emissions with Emphasis on Central Sewage Facilities.
Atmospheric Environment 9:941-944, 1975.
224
-------
154. Farrell, J. B., H. 0. Wall and b. A. Kerdolff. Air Pollution from
Sewage Sludge Incinerators. Unpublished Report. U.S. Environmental
Protection Agency, Cincinnati, Ohio 1978.
155. Jacknow, J. Environmental Aspects of Municipal Sludge Incineration.
In: Acceptable Sludge Disposal Techniques: Cost, Benefit, Risk,
Health, and Public Acceptance, 47-51, Information Transfer, Inc.,
Rockville, Maryland, 1978,
156. Practical, Available Technology (PAT) Report. Sludge Incineration.
Env. Sci. and Technol.lO(12):1080-82, 1976.
157. Dew!ing, R. T., R. N. Manganelli and G. T. Baer, Jr. Fate and Behavior
of Selected Heavy Metals in Incinerated Sludge. Jour. WPCF 52(10)=2552-2557
1980.
158. Gray, D. H. and C. Penessis. Engineering Properties of Sludge Ash.
Jour. WPCF 44(5):847-858, 1972.
159. Havens and Emerson. Proposed Sludge Management Plan. Metropolitan
District Commission, Boston, 1973.
160. U.S. Environmental Protection Agency. Environmental News, August 23,
1976.
161. U.S. Environmental Protectior, Agency. Sewage Sludge Incineration.
Report of the Task Force for the Office of Research and Monitoring.
EPA-R2-72-040, Washington, D.C., U.S. Environmental Protection Agency,
PB-211 323, Springfield, Virginia, National Technical Information
Serv.nce, 1972.
162. Califronia Administrative Code: Title 23. Chapter 3, State Water Re-
sources Control Board, Subchapter 15, Waste Disposal to Land, March
10, 1972 (updated on June 7, 1980).
163. Palo Alto Finds Gold in its Sludge Ash. Water Pollution Control Fed.
Highlights 17(2}:2, 1980.
164. Brown and Caldwell Consulting Engineers. Solid Waste Resource Re-
covery Full Scale Test Report. Central Contra Costa Sanitary District,
California, Volumes One and Two, March 1977.
165. Sieger, R. B. and B. D. Bracken. Sludge, Garbage May Fuel California
Sewage Plant. The American City and County, p. 37, January 1977.
166. Huffman, G. L. and W. W. Liberick, Jr. EPA's R & D Program in Pyrolytic
Conversion of Waste to Fuel Products. In: Solid Wastes and Residues.
Conversion by Advanced Thermal Processes. J. L. Jones and S, B. Radding,
Eds., ACS Symposium Series 76, American Chemical Society, 1978.
225
-------
167. Moses, C- T., K. W. Young, G. Stern and J. B. Parrel. Co-Disposal
of Sludge and Refuse in a Purox Converter. In: Solid Wastes and
Residues. J. L. Jones and S. B. Radding, Eds., ACS Symposium Series
76, American Chemical Society, 1978.
168. Galandak, 0. and M. Racstain. Design Considerations for Pyrolysis
of Municipal Sludge. Jour. WPCF 51(2):370-376, 1980.
169. Metz, W. C., J. Shyer and K. Edgecomb. Preliminary Assessment of
the Prospects for Use of Refuse - Derived Fuel in Maryland. Brookhaven
National Lab., Upton, N.Y., February 1979, 83 pp.
170. Mazima, T., K. Tadao, M. Naruse and M. Hiraoka. Studies on the
Pyrolysis Process of Sewage Sludge. Prog. Water Tech. 9:381-396, 1977.
171. Olexsey, R. A. Pyrolysis of Sewage Sludge. Proceedings of the 1975
National Conference on Municipal Sludge Management and Disposal,
Anaheim, California, August 18-20, 1975.
172. Lewis, F. M. Sludge Pyrolysis for Energy Recovery and Pollution
Control. Proceedings of the Second National Conference on Municipal
Sludge Management and Disposal. Anaheim, California, August 18-20,
1975.
173. U. S. Environmental Protection Agency. Proceedings of Fifth United
States/Japan Conference on Sewage Treatment Technology, Chapter 4,
EPA-600/9-77-027, April 18-22, 1977, Tokyo, Japan.
226
-------
APPENDIX A
Musjc-gon_ 6/4/79
Date
Monthly Illness History
T.D. Number
1. Have you been sick in any way in the past month? Yes _ No _
If "yes" then answer -the following:
a. Briefly describe the illness, when it occurred, and how many days it
lasted:
b. Indicate which of the following you had:
Yes No * days
1. Fever (list highest recorded temperature)
2. Chills
3. Eye inflamaation (red eye)
4. Runny nose ____
5. Sneezing _____ _ ______
6. Sore throat
7. Coughing
8. Shortness of breach
9. Chest pain
10. Pneumonia
11. Nausea
12. Vomiting
13. Stomach pain _____ _____ ^^^^
14. Diarrhea '
15. Dermatitis (skin rash or infection) _____
227
-------
APPENDIX A (continued)
- 2 -
2. Have you seen a doctor in the past month? Yes No_
If "yes" then answer the following:
a. What doctor did you see?
b. What was his diagnosis?
c. Did he prescribe any medicine?
3. Have you taken any medicine such as aspirin, cold remedies, allergy or
diarrhea medicines in the past month? Yes No
If "yes" then answer the following:
a. What were the medicines?
b. When did you take them (approximate dates)_
c. How many days did you take them?
4. Has any of the following changed in the past month - job?, address?, telephone
number?, household size?, etc. Yes No
If so, please specify_
5. How many times have your clothes become soaked or heavily soiled at work
during the past month?
6. Have we obtained all your specimens (blood, throat and rectal swabs) this
month? Yes No
228
------- |