September 1975
Environmental Protection Technology Series
A VIRUS-IN-WATER STUDY OF
FINISHED WATER FROM
SIX COMMUNITIES
Municipal Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development,
U.S. Environmental Protection Agency, have been grouped into
five series. These five broad categories were established to
facilitate further development and application of environmental
technology. Elimination bf traditional grouping was consciously
planned to foster technology transfer and a maximum interface in
related fields. The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ENVIRONMENTAL HEALTH EFFECTS
RESEARCH STUDIES series. This series describes projects and studies
relating to the tolerances of man for unhealthful substances or
cdhditions. This work is generally assessed from a medical view-
point, including physiological or psychological studies. In addition
to toxicology and other medical specialities, study areas include
biomedical instrumentation and health research techniques utilizing
animals - but always with intended application to human health measures
This document is available to the public through the National
Technical Information Service, Springfield, Virginia 22151.
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EPA-600/1-75-003
September 1975
A VIRUS-IN-WATER STUDY OF
FINISHED WATER FROM SIX COMMUNITIES
by
Elmer W. Akin
David A. Brashear
Norman A. Clarke
Water Quality Division
Program Element No. 1CA046
HEALTH EFFECTS RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
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DISCLAIMER
This report has been reviewed by the Health Effects
Research Laboratory, U.S. Environmental Protection Agency,
and approved for publication. Mention of trade names or
commercial products does not constitute endorsement or
recommendation for use.
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FOREWORD
Man and his environment must be protected from the adverse effects of
pesticides, radiation, noise and other forms of pollution, and the unwise
management of solid waste. Efforts to protect the environment require a
focus that recognizes the interplay between the components of our physical
environment—air, water, and land. In Cincinnati, the Environmental
Research Center possesses this multidisciplinary focus through programs
engaged in
• studies on the effects of environmental contaminants on
man and the biosphere, and
• a search for ways to prevent contamination and to recycle
valuable resources.
The Health Effects Research Laboratory conducts studies to identify
environmental contaminants singly or in combination, discern their
relationships, and to detect, define, and quantify their health and
economic effects utilizing appropriate clinical, epidemiological,
toxicological, and socio-economic assessment methodologies.
Enteric viruses are an environmental contaminant through the sewage
system. This continuing study was designed to determine if human enteric
viruses could be found in treated drinking water and to field test virus
concentrating equipment and procedures as they were improved by ongoing
research in the Health Effects Research Laboratory.
R. J. Garner
Acting Director
ill
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ACKNOWLEDGMENTS
We thank Charles Mayhew and Melvin Sparks for collecting the samples
and Theadore Ericksen for performing the bacteriological tests.
iv
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INTRODUCTION
Six sites in three States were selected for a virus-1n-water study,
with the assistance of EPA regional staff and State health department
(or State EPA) personnel. The sites selected were: (1) Columbus, Ohio
(Dublin Road Plant). (2) Sidney, Ohio, (3) Seymour, Indiana, (4) Muncie,
Indiana (5) St. Joseph, Missouri and (6) Kansas City, Missouri. The
primary criterion for selection of sites was based on a treatment plant's
use of surface source water having domestic contamination as indicated
by high fecal coliform counts. An effort was also made to select sites
that used conventional flocculation or softening procedures. In addition,
plants of varying sizes were chosen as indicated by water output volumes.
The study had a two fold purpose: (1) to determine if human enteric
viruses could be detected in treated finished water with the equipment
and procedures presently available, and (2) to field test virus-sampling
equipment and procedures as they were improved by ongoing research in
this laboratory.
MATERIALS AND METHODS
The types of treatment and the fecal coliform densities of the source
waters at the six sites are shown in Table 1. Plant capacity ranged from
3
5,600 to 795,000 m per day (1.5 to 210 million gallons per day) and
treatment included both conventional alum flocculation and lime-soda
softening. The range and geometric means of the fecal coliform densities
in the source waters indicated that relatively heavy pollution was
occurring intermittently at the Ohio and Indiana sites and almost constantly
1
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at the Missouri sites.
A flow-through virus-adsorbent system was used throughout this study.
Water samples were adjusted to pH 3.5 and in the initial samples, divalent
cations were added to enhance virus adsorption (it was later shown that
the addition of cations did not enhance virus recovery). Sodium thio-
sulfate was also added to neutralize residual chlorine in the water
samples. After the desired volume of water had been processed, the virus
adsorbant was then transported to the laboratory on ice and eluted with
one liter of eluting fluid. A secondary concentration step was conducted
to further reduce the sample (eluate) volume. The reconcentrated sample
(10-30 ml) was then distributed equally among four cell types: (1) primary
African green monkey kidney (AQMK), (2) primary human embryonic kidney (HEK),
(3) continuous porcine kidney and (4) continuous monkey kidney (VERO).
These inoculated cell monolayers were observed for cytopathic effects
(CPE) for 14 days; if the results were negative, two consecutive blind
passages were conducted before declaring a sample negative for viruses.
Cell monolayers manifesting CPE were confirmed as viral CPE and the
serotype identified by serum neutralization test.
Sampling was begun in November 1973 with the objective of collecting
and processing 380 liters (100 gal) of water per sample. The first 11
samples were collected using a 0.45-vm porosity cellulose nitrate
cylindrical filter (Millitube) as the virus adsorbent. The filter was
eluted with 5x nutrient broth pH 9.0 and this eluate was reconcentrated
by an aqueous polymer 2-phase procedure. Premature plugging, eluate
leakage and other problems led to the discontinuation of this procedure.
Laboratory experimentation had previously indicated that a stack of three
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293HTDH cellulose nitrate membrane disks (8, 1,2 and ,45-nm porosities)
was a better system. Beginning with sample number 12, this system
replaced the Millitube as the primary adsorbent. In addition, the
nutrient broth eluent was replaced with glycine buffer pH 11.5 (later
reduced to pH 11.1) and the polymer 2-phase reconcentration procedure
was replaced with a second virus adsorbent system^a stack of three
small diameter (47 iron) epoxy^fiberglass^asbestos disks of 5, 1 and .45-ym
porosity. Premature plugging occurred on occasions with the 293-mm
membrane disk.
Further experimentation in our laboratory with an epoxy-fiberglass
filter tube of 8.0-ym porosity had previously indicated that a larger
sample volume (1,900 liters) could be collected with a greater poliovirus
recovery sensitivity when three filter tubes in parallel were used as the
virus adsorbent1. This unit also had advantages in size and ease of
handling and was adopted in July 1974 as the primary virus adsorbent
system. Premature plugging has not been experienced with this virus
adsorbing filter.
A positive control system was included in this study to keep a
continual check on the virus recovery capability of the procedure.
Ampules of sterile nutrient broth were prepared in the laboratory and
low levels of selected enteric virus serotypes were introduced into a
number of these ampules. The ampules were then coded, randomized by
number and the content of a single ampule was introduced into each field
sample as it was being processed. Neither the sample collecting team
nor the person "reading" the cell culture for CPE knew which ampules
contained a virus. The project officer monitored the distribution of
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the positive controls and also conducted the serological test that
confirmed the recovery of a positive control when CPE was observed in
the cell culture system. The selection of different virus serotypes
as positive controls provided an opportunity to evaluate the virus
detection efficiency of the procedure for representatives of the
various subgroups within the enteric virus group.
Samples were collected to test for the presence of coliform bacteria
at the time that water samples were taken for virus testing. This
procedure was followed to attempt to relate any virus isolations to the
presence of the coliform indicator of microbial pollution. The presence
of coliform bacteria was determined by the membrane filter procedure
with 0.1, 1.0 and 5.0 liters of sample and by two experimental procedures.
The first experimental procedure was a modified MPN test in which membrane
filters were used and up to 5.5 liters of water were examined; the other
was a large-volume sampler (LVS) technique that utilized epoxy-fiberglass
filter tubes whereby 380 liters of water were filtered and the coliform
densities determined by the MPN procedure. The LVS was under development
at the time of this study and was not available for field use until June
1974. The inclusion in this study of 1 and 5 liter sample volumes for
the membrane filter procedure and the two experimental procedures reflected
the concern that 100-ml standard portions may be insufficient volumes to
examine if the coliform test is to be a satisfactory indicator of viral
contamination of a water supply. If viruses had been isolated from the
finished water samples taken in this study, an attempt would have been
made to correlate these isolations with coliform densities derived from
the three bacteriological procedures.
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RESULTS
Between November 1973 and February 1975 seventy nine samples were
collected from the six water treatment plants at a point after final
treatment and disinfection. Pertinent data on the samples collected at
each of the six sites are shown in Tables 2 through 7- Turbidities were
generally very low (<1.0 unit) and free chlorine residuals were over 1.0
mg/1 in most samples from all sites except Kansas City which added ammonia
to produce chloramines. All samples were alkaline and approached pH 10
at the plants using a lime softening procedure (see Table 1). Difficulties
were experienced in processing 15 of the 79 samples. The processing of
these samples is recorded as unsatisfactory in the tables because it was
felt that virus detection could have been significantly compromised. Of
the remaining 64 samples, eight received the positive control viruses as
a check on the virus recovery procedure. The control viruses were recovered
in six of the eight samples (Table 8). Two virus serotypes (Reo 1 and
Echo 27) were not recovered when introduced at a concentration of one
infectious unit per 19 liters. Other viruses were recovered at this
low level and studies are now underway to determine the cause of our
failure to recover these two viruses. The remaining 56 samples represent
the study samples from the six sites.
Cell culture changes indicative of viral replication were observed
in AGMK cells that had received two of the study samples. These two
isolates were subsequently identified, by an independent laboratory and
confirmed in our laboratory, as a monkey papovavirus (SV 40) that is
commonly indigenous to primary monkey kidney cells. None of the other
-------
three cell types used manifested CPE, It was concluded that these two
viral isolates were cell culture contaminants and that therefore, no
viruses had been isolated from any of the study samples.
The source water quality at each of the six sites was very poor at
certain times during the study period. At least one sample from each site
exceeded the maximum fecal coliform densities recommended for source water^
The two Missouri sites used the Missouri River as their water source, a
river that is well recognized as having a heavy microbial pollution load.
Fecal coliform densities were consistently high at the intake of these
two water treatment plants (Table 1).
Bacteriological tests for coliform organisms were conducted on 71
(37 with the LVS) of the 79 finished water samples. Coliform-group
organisms or fecal coliforms were detected in 25 of these samples by
at least one of the three procedures used (Table 9). Using the membrane
filter procedure with up to 5 liters of sample, coliforms were detected
in seven samples, one of which also yielded fecal coliforms. Using the
modified MPN procedure with 5.5 liters of sample, coliforms were detected
in 11 samples, two of which also yielded fecal coliforms. Using the
LVS and examining 380 liters of water, coliforms were detected in 17
samples, eight of which also yielded fecal coliforms. In only three
samples (all from Seymour) were coliforms detected by all three procedures.
Since these samples were collected and partially processed in the field
under somewhat adverse conditions, it is possible that some of the positive
samples represent'contamination. In all cases, the coliform densities
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were extremely low and all samples met the 1962 U.S. Public Health
Service Drinking Water Standard.
DISCUSSION
The health risk of human viruses in treated water supplies in this
country has not been established. In fact, the studies by this
laboratory are believed to be the first attempt to use a sensitive,
large-volume virus recovery procedure to examine drinking water for
the presence of viruses at plants that use domestically polluted surface
water as a source? In the absence of background surveillance data, the
volume of each sample, the frequency of sampling and the number of samples
have not been established for determining the viral quality of a water
supply. The virus recovery efficiency of our procedure had been evaluated
in the laboratory with sample volumes only up to 1,900 liters (500 gal).
Therefore, we made no attempt to exceed this volume with the field samples.
The frequency of sampling and the total number of samples taken was
primarily determined by staff and facilities available for the study and
distance of the site from the laboratory. It was felt that more than one
sample was required from each site, because of the fluctuations in the
quality of source waters and the variations in the day to day operation
of water treatment plants, before a definitive conclusion could be drawn
as to the virological quality of the water.
In this study, three to 14 samples per site were collected and
successfully processed. At this point in the study of the virus-in-water
question, we believe that fewer than five samples are an insufficient
-------
number to evaluate a water supply. Therefore, sampling will continue at
the Kansas City site (4 satisfactory study samples collected) and the
St. Joseph site (3 satisfactory study samples collected) before an
evaluation will be made. Nine to 14 samples were successfully processed
from each of the Ohio and Indiana sites. Analysis of the waters at the
time of sampling indicated that treatment for removal of pathogenic
microorganisms was very good, i.e., high free chlorine residuals and low
turbidities. Therefore, in the absence of virus isolations, sampling at
these four sites will not be continued as more appropriate sites are
located.
The isolation of fecal coliforns from six of the Seymour, Indiana,
samples with the LVS is of special interest (Table 9). The significance
of low numbers (7 to > 43/1,000 liters) of these organisms in treated
drinking water with free chlorine residuals >1.0 mg/1 cannot be evaluated
in the absence of virus isolations. Continued sampling with the LVS
and a thorough evaluation of its recovery efficiency is certainly in
order and will be carried out.
The results of this study of finished waters produced by conventional
treatment methods from surface waters of a rather poor quality have shown
that if viruses were present in these waters, the numbers were below the
detectable level of a sensitive virus recovery procedure. Laboratory tests
have shown that the sensitivity of the virus recovery procedures used in
this study was such that poliovirus could consistently be detected at
contamination levels of 3 to 5 units per 380 liters when 1,900 liters
were sampled!* Through the evaluation of positive controls, field tests
have recovered six other members of the enteric virus group at concentrations
8
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of 100 to 1,000 virus units per 380 to 1,900 liters. Therefore, we
assume that if viruses were present in the study samples, the concentra-
tion was below one virus unit per 3.8 liters (gal) of finished water.
The fact that these water treatment systems were challenged by very
poor source water on numerous occasions and yet produced a finished
water in which no viruses could be recovered seems to confirm the
adequacy of good conventional treatment for virus removal.
In light of the failure to recover virus from up to 1,900 liters
(500 gal) of finished water from these sites, it now seems desirable to
study water plants or water systems that have compromised or completely
omitted conventional treatment of a source water known to have recently
received domestic waste. We are presently seeking such sites. Documen-
tation of the level of viral contamination in the source water of some
sites may also be desirable as a basis for evaluating the viral removal
efficiency of the treatment systems. Procedures for collecting and
processing large volumes of untreated surface water for virus isolation
are presently being evaluated in our laboratory.
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REFERENCES
1. Clarke, N. A., Hill, W. F. Jr., Jakubowski, W.; "Detection of Viruses
in Water: Tentative Standard Method"; AWWA Tech. Conf. Proc., Dec. 3
and 4, 1974, Dallas, Texas (1975).
2. Water Quality Criteria 1972, Report of the Committee on Water Quality
Criteria, National Academy of Science and National Academy of
Engineering, Washington, D.C. pg 57 and 58 (1973).
3. Clarke, N. A., Akin, E. W., Liu, 0. C., Hoff, J. C., Hill, W. F. Jr.,
Brashear, D. A., and Jakubowski, W.; "Virus Study for Drinking-Water
Supplies"; Jour. AWWA 67; 192-197 (1975).
4. Hill, W. F- Jr., Jakubowski, W., Akin, E. W., Clarke, N. A.; "Detection
of Virus in Drinking Water: Sensitivity of the Tentative Standard
Method"; Abs. Annual Mtn.-1975, Am. Soc. for Microbiol. (1975).
10
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Table 1. TREATMENT PROCESSES EMPLOYED AND FECAL COLIFORM
DENSITIES OF SOURCE WATER AT SIX STUDY SITES
Item
Treatment:
Potassium permanganate
Prechlorinatlon
Presedimentatlon
Ammonia
Alum
Lime
Carbon
Polymer
Lime-soda softening
Flocculation
Sedimentation
Chi ori nation
Recarbonation
Sand filtration
Post chlorination
Contact time (hr)
Capacity/mean output (10,000 m3/day)
Fecal conform MPN/100 ml of
source water (range)
Geometric mean
Columbus
X
X
X
X
X
X
X
X
X
26
26.5/13.6
38-3,000
280
Sidney
X
X
X
X
X
X
X
X
7
1.4/1.1
40-2,800
263
Treatment
Site
Muncie Seymour
Xa
X
X
X
X
X
X
X
X
8
6.0/4.9 0
50-2,200
210
X
X
xa
X
X
X
X
14
.56/0.45
11-14,000
536
Kansas City
X
X
X
X
xa
Xa
X
X
X
X
X
X
22
79.5/41.5
1,700-4,300
3,010
St. Joseph
Xa
Xd
X
X
X
X
X
X
24
11/5
280-7
1,890
.3
,000
a Added as needed
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Table 2. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT COLUMBUS, OHIO
Date Volume Turbidity
collected collected (liters) (FTU)
12/5/74
1/21/74
3/11/74
3/25/74
4/16/74
5/1/74
5/15/74
5/29/74
6/26/74
7/10/74
8/14/74
9/10/74
10/2/74
10/16/74
11/6/74
265
380
380
380
380
380
380
380
380
1,900
1,900
1,900
1,900
1,900
1,900
.08
.49
.07
.09
.03
.04
.05
.05
.07
.28
.72
.04
.09
.04
.02
Re
Residual chlorine (mg/1) sa
Free
2.6
1.5
1.5
1.4
1.4
1.4
1.8
1.5
1.7
1.4
1.4
1.2
1.6
1.3
1.3
Combi ned
.38
.45
.40
.35
.20
.15
.00
.32
.40
.27
.20
.20
.20
.10
.40
pH pr
9.5
9.4
9.3
9.5
9.2
9.6
9.8
10.1
9.6
9.6
10.2
9.9
10.1
10.2
10.1
isult of
imple
•ocessing
S
S
S
PC
S
S
S
S
S
S
S
S
S
S
S
S, successful; PC, positive control.
12
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Table 3. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT SIDNEY, OHIO
00
Date Volume Turbidity
collected collected (liters) (FTU)
11/26/73
1/16/74
3/12/74
3/26/74
4/17/74
4/30/74
5/14/74
5/28/74
6/25/74
9/11/74
10/1/74
10/15/74
11/5/74
570
380
380
380
380
380
380
380
380
1,900
1,900
1,900
1,900
.00
.02
.03
.02
.03
.02
.02
.01
.05
.00
.08
.04
.05
Result of
Residual chlorine (mg/1) sample
Free Combined pH processing
2.0
1.3
1.4
0.9
1.3
1.4
1.6
2.2
1.9
1.3
1.9
1.6
1.4
1.1
1.8
0.6
1.0
0.7
0.5
1.4
0.5
1.0
0.4
0.9
0.3
0.8
9.0
8.8
9.3
9.3
8.8
9.2
9.7
9.7
9.2
9.7
9.8
9.1
9.9
US
S
S
US
S
PC
S
S
S
S
PC
S
PC
a S, successful; US, unsuccessful; PC, positive control.
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Table 4. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT MUNCIE, INDIANA
Date Volume Turbidity
collected collected (liters) (FTU)
12/18/73
1/14/74
2/26/74
3/19/74
4/10/74
4/24/74
5/8/74
5/22/74
6/5/74
7/3/74
7/17/74
7/24/74
9/4/74
9/26/74
10/9/74
10/31/74
11/12/74
12/3/74
285
380
380
380
380
285
170
380
380
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
0.26
0.04
0.76
0.41
0.28
0.14
0.51
0.10
0.08
0.18
0.36
0.11
0.33
0.56
0.71
1.00
0.30
0.56
Result of
Residual chlorine (mg/1) sample
free Combined pH processing
1.2
1.8
1.6
1.9
1.5
2.0
1.0
1.6
1.5
1.0
1.4
1.7
1.4
1.3
2.0
1.1
1.7
2.2
0.1
0.3
0.3
0.5
0.4
0.2
0.4
0.2
0.1
0.3
0.3
0.3
0.3
0.1
0.2
0.2
0.3
0.1
7.7
7.4
7.1
7.4
7.3
7.4
7.8
7.6
7.7
7.2
8.4
8.2
8.5
8.9
9.2
8.1
8.4
7.7
S
US
S
S
S
us
us
S
S
S
S
S
S
S
us
S
S
PC
S, successful; US, unsuccessful; PC, positive control.
14
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Table 5. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT SEYMOUR, INDIANA
Date Volume Turbidity
collected collected (liters) (FTU)
11/27/73
1/6/74
2/12/74
3/18/74
4/9/74
4/23/74
5/7/74
5/21/74
6/4/74
7/2/74
7/16/74
7/23/74
8/8/74
9/5/74
9/25/74
10/8/74
10/30/74
11/11/74
12/2/74
1/4/75
1/15/75
2/12/75
2/13/75
95
240
380
380
380
380
380
380
380
1,900
1,900
1 ,900b
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
1,900
.60
.14
.36
.19
.43
.08
.30
.34
.83
.11
.14
.03
.03
.36
.08
.38
.40
.10
.44
1.50
1.00
.45
.44
Residual
Free
0.4
0.7
1.0
1.0
0.7
0.8
0.7
0.4
0.2
0.4
0.3
2.5
1.7
2.4
1.5
1.8
1.6
1.7
1.9
1.7
2.0
2.4
2.7
Re:
chlorine (mg/11 sai
Combined
0.2
0.1
0.3
0.2
0.3
0.3
0.2
0.2
0.0
0.5
0.0
0.3
0.3
0.2
0.3
0.1
0.2
0.3
0.1
0.4
0.1
0.1
0.1
pH pri
7.3
7.6
7.6
7.4
7.2
7.6
8.2
7.5
7.2
7.2
7.8
8.2
7.9
8.7
9.1
8.8
8.0
8.3
7.3
7.0
7.1
8.1
8.4
suit of
nple
ocessing
S
US
S
S
S
PC
S
S
S
US
us
us
S
S
S
us
us
S
S
us
S
PC
S
* Successful; US, unsuccessful; PC, positive control.
" Began sampling at new site in plant.
15
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Table 6. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT KANSAS CITY, MISSOURI
Date
collected
12/12/73
6/12/74
7/31/74
8/21/74
9/18/74
11/20/74
Volume
collected (liters)
270
380
1,140
1,900
880
1,900
Turbidity
(FTU)
.05
.07
.03
.02
.03
.71
Residual
Free
.03
.00
.02
.00
.00
.00
chlorine (mg/ll
Combined
1.2
0.6
0.8
1.0
1.0
0.8
Result of
sample
pH processing
9.5 S
10.5 PC
10.2 US
9.7 S
9.9 S
9.8 S
a S, successful; US, unsuccessful; PC, positive control.
Table 7. VOLUME, WATER QUALITY DATA, AND PROCESSING RESULTS OF
SAMPLES COLLECTED AT ST. JOSEPH, MISSOURI
Date
collected
12/12/73
6/12/74
7/30/74
8/20/74
Volume
collected (liters)
260
380
1,900
1,900
Turbidity
(FTU)
.28
.09
.63
.08
Residual
Free
2.0
1.9
1.7
1.9
chlorine (mg/1)
Combined
0.1
0.3
0.3
0.2
PH
7.9
8.0
8.0
8.4
Result of
sample
processing
S
US
S
S
a S, successful; US, unsuccessful.
16
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Table 8. VIRUS INOCULATED INTO RANDOMLY SELECTED SAMPLES
AS A CONTROL ON THE VIRUS RECOVERY SENSITIVITY
OF THE SAMPLING! PROCEDURE
Sampling
site
Columbus
Seymour
Sidney
Kansas City
Sidney
Sidney
Muncie
Seymour
Sample
volume (liters)
380
380
380
380
1,900
1,900
1,900
1,900
Date
3/25/74
4/23/74
4/30/74
6/12/74
10/1/74
11/5/74
12/3/74
2/12/75
Virus
Type
Echo 24
Coxsackie
Adeno 15
Echo 13
Reo 3
Coxsackie
Echo 27
Polio 1
Input
Amount^
100
A21 1 ,000
100
100
100
B6 100
100
160
Result of
isolation
procedure b
R
R
R
R
NR
R
NR
R
The poliovirus was quantitated by the plaque method; all others
by the tissue culture Infectious Dose 50% method.
b R, virus recovered; NR, virus not recovered
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Table 9. POSITIVE COLIFORM/FECAL COLIFORM FINDINGS FOR
FINISHED WATER SAMPLES FROM SIX STUDY SITES
Sample site
and
date collected
Membrane filter3
Total Fecal
coliforms coliforms
Modified MPIT
Total Fecal
coliforms coliforms
Large volume sampler
Total Fecal
coliforms coliforms
Columbus:
12/5/73
1/21/74
10/2/74
11/6/74
Sidney:
9/11/74
10/8/74
11/5/74
Muncie:
7/3/74
9/4/74
9/26/74
10/31/74
Seymour:
7/23/74
9/5/74
9/25/74
10/8/74
10/30/74
11/11/74
12/2/74
1/14/75
1/15/75
2/12/75
2/13/75
Kansas City:
12/12/73
6/12/74
St. Joseph:
6/12/74
+
+
+
+
+
ND°
ND
ND
ND
Up to 5 liters were tested; negative findings indicate a value <0.02; the largest
positive value was 0.12 organism/100 ml.
b 5.5 liters were tested; negative findings indicate a value <0.02 organism/100 ml;
the largest positive value was 0.23 organism/100 ml.
c 380 liters were tested; negative findings indicate a value <0.00059 organisms/100 ml;
the largest positive value was >0.0043/100 ml.
d ND, not done.
18
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
, REPORT NO.
EPA-600/1-75-003
2.
3. RECIPIENT'S ACCESSIOONO.
4. TITLE AND SUBTITLE
A VIRUS-IN-WATER STUDY OF FINISHED WATER FROM
SIX COMMUNITIES
5. REPORT DATE
September 1975 (Issuing Date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Elmer W. Akin, David A. Brashear, and
Norman A. Clarke
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Health Effects Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
10. PROGRAM ELEMENT NO.
1CA046; ROAP 21 APX; Task 08
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Same as above
13. TYPE OF REPORT AND PERIOD COVERED
In-house
14. SPONSORING AGENCY CODE
EPA-ORD
15. SUPPLEMENTARY NOTES
16. ABSTRACT
Fifty-six finished water study samples up to 1900 liters were collected
and successfully processed for virus isolations from six communities. Eight
additional samples were inoculated with low levels of virus as a check (positive
control) on the sensitivity of our procedure. Six of the eight positive control
viruses were recovered. Two virus serotypes were not recovered at an input level
of one infectious unit per 19 liters. No viruses were isolated from the study
samples. Bacteriological tests with experimental large volume procedures showed
that coliform bacteria were present in 25 of 71 samples. The coliform densities
were very low and in all cases were within the limits of the 1962 USPHS Drinking
Water Standards.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
COSATl Field/Group
Water treatment
Microorganism control (water)
Viruses
Coliform bacteria
Potable water
Occurrence
Enteric viruses
13B
6M
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
UNCLASSIFIED
21. NO. OF PAGES
23
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
19
•ftUSGPO: 1975 — 657-695/5310 Region 5-11
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