IN WASTE,
RENOVtATID,
A MO OTHER WATERS
1976
jJTERATUMABSTRACTS
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0.
U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Research and Development
Environmental Research Center
Cincinnati, Ohio
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EPA-600/9-77-028
August 1977
VIRUSES IN WASTE,
RENOVATED,
AND OTHER WATERS
Editor: Gerald Berg, Ph.D.
Editorial Assistant: F. Dianne White
1976
BIOLOGICAL METHODS BRANCH
ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
OFFICE OF RESEARCH AND DEVaOPMENT
ENVIRONMENTAL RESEARCH CENTER
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO
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AUTHOR AND SUBJECT INDEXES
VIRUSES IN WASTE, RENOVATED, AND OTHER WATERS
now contains an Author Index and a Subject Index. Both listings
he found at the end of this volume. The Greek letters that ap-
pear at c\’e ach abstract are the coding for the Subject Index. An
explanation for their meanings is given on the first page of that
index.
DISCLAI MER
This report has been reviewed by the Environmental Research
Center—Cincinnati, USEPA, and approved for publication. Mention
of trade names or commercial products does not constitute en-
dorsement or recommendation for use.
II
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1976
VIRUSES IN WASTE, RENOVATED, AND OTHER WATERS
7T O
Aizen, MS., Pille, ER. (1976). Role of Food Products in the
Spread of Viral Infections. ZH MIKROBIOL EPIDEMIOL IMMU-
NOBIOL, O(3):12-7. Russian.
Outbreaks of shellfish-borne hepatitis A, and outbreaks of
other food-borne viral diseases in which water played some role
are briefly discussed within a review of food-borne viral diseases.
Akin, E.W., Hill, W.F., Jr., Cline, GB., Benton, W.H. (1976). The
Loss of Poliovirus 1 Infectivity in Marine Waters. WATER RES,
1O(1):59-63.
Three logs of infectivity of poliovirus 1 were lost in 5 to 6 days
at 24 C in raw, filter-sterilized, and autoclaved marine waters. This
loss of infectivity apparently did not result from aggregation of
viruses or from adsorption of viruses to containers. Two-com-
ponent curves often occurred with the loss of virus; a resistant
population in the seed virus that might account for such curves
was not found.
Loss of infectivity occurred when the virus was suspended in
artificial seawater of 1, 10, and 20 g/kg salinity. No explanation for
this loss, other than inactivation of the virions, was found. The
specific component(s) of marine water responsible for the inac-
tivation was not identified.
1
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A
Albano, A., De Donato, S. (1974). A Simple Procedure for De-
tecting Viruses in Sewage. BOLL 1ST SIER, 53(1):16-23. Italian.
A procedure that consisted of passing settled sewage through
filter paper and eluting adsorbed and trapped viruses from the tnt-
urated paper with 10% calf serum in borate buffer (pH 9) was
about equal in effectiveness to the gauze pad procedure and con-
siderably superior to the polyelectrolyte procedure for recovering
viruses from sewage.
Ii
Alexandre, D., Gevaudan, P., Charrel, J., Gulian, C., Mallet, M.N.,
Blancard, A. (1975). Study of the Survival of Microorganisms in
Wastewaters During Purification by Physico-Chemical Treatment.
TECH EAU ASSAINISSEMENT, 345:31-8. French.
From 37 to 94% of seeded poliovirus 3 was removed from
sewage by flocculation with alum A 1 2 (S0 4 ) 3 at pH 9. Flocculation
at pH 11.5 removed 99.87% of the seeded virus or more. The
quantity of viruses removed remained unchanged when carbon
was added to the flocced sewage.
The alum concentration in all experiments was 200 gm/rn 3 .
The pH levels were achieved with the addition of lime [ Ca(OH) 2 ].
Activated carbon when used was applied in a concentration of 150
gm/rn 3 at pH 9 and at a concentration of 100 gm/rn 3 at pH 11.5.
Bachmann, P.A. Epidemiological and Epizootiological Significance
of Pathogenic Microorganisms in Surface Water and Sewage. In
“Munich’s Contributions to Sewage, Fishery and Stream Biology,
Vol. 26, Water for Land Reclamation Areas.” Oldenbourg Verlag,
Munich, West Germany (1975), 155-65. German.
The transmission of viral diseases by water is reviewed in re-
lation to the frequency and types of viruses found in water, the
survival of viruses in surface waters and sewages, and what con-
stitutes the minimal infective dose for viruses.
A
Belfort, G.rn, Rotem, V., Katzenelson, E. (1976). Virus Concentration
Using Hollow Fiber Membranes. II. WATER RES, 1O(4):279-84.
Poliovirus 1 was concentrated in asymmetric polysuiphone
hollow fiber membranes. Average recovery efficiencies of 61 and
77% were achieved from 5- and 50-liter quantities of deionized wa-
ter, respectively, and an average recovery efficiency of 52°Io was
achieved from 50 liters of tap water. From 22 to 45% of the recov-
eries were obtained from filter backwash.
In earlier experiments with symmetric semi-dense cellulose
acetate hollow fibers, an average of 84% of the poliovirus was re-
covered from 5 liters of deionized water, 36% from filter backwash.
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Effectiveness of the membranes was unaffected by the initial
virus concentrations which spanned a five-log range.
The asymmetric membranes exhibited higher virus rejections
before they were backwashed, a higher water permeation coeff I-
dent, and suffered less penetration of virus into the membranes
than the symmetric membranes did.
Bell R.G. (1976) The Limitation of the Ratio of Fecal Co/if orms to
Total Co/iphage as a Water Pollution Index. WATER RES,
1O(8):745-8.
The ratios of fecal coliforms (MPN) to coliphage (of
Escherichia co/i B, ATCC 11303-1) in raw sewage, sewage lagoon
effluent, and in river water were 87:1, 4.2:1, and 0.15:1, respectively.
The ratio of fecal coliforms to coliphage in stored raw sewage de-
creased from 87:1 to about 1:1 within 7 days at 20 C and within 28
days at 4 C.
The ratio of fecal coliforms to coliphage is not a reliable index
of fecal pollution because the ratio is affected by environmental
conditions.
Berry 1 S.A., Noton, BG. (1976). Survival of Bacteriophages in Sea-
water. WATER RES, 1O(4):323-7.
In laboratory studies at 4 C and 12 C, coliphage T2 titers were
stable in artificial seawater for 48 days. In unpolluted seawater,
about 90% of the phage was inactivated in 48 days at 4 C, and
about 95% of the phage was inactivated in 48 days at 12 C. In
heavily polluted seawater, about 99% of the phage was inactivated
in 48 days at 4 C and more than 99.99% was inactivated in 48 days
at 12C. In 14 days, dialysed in tubing against normal seawater at
12 C, more than 99.99999% of the phage was inactivated in pollut-
ed seawater, and more than 9 9.9% of the phage was inactivated in
normal seawater.
The phage survived longer in centrifuged, membrane-filtered,
or autoclaved seawater than in natural seawater, probably because
organic and microbial material were removed from or destroyed in
the centrifuged, membrane filtered, and autoclaved waters.
The phage, in artifical seawater or in polluted seawater (in di-
alysis bags), was inactivated more rapidly in shallow (0-8.7 feet)
than in deep (4.6-15 feet) seawater, possibly the result of exposure
to sunlight.
Coliphage T2 survived longer in seawater in the laboratory
than in seawater (in dialysis tubes) in a bay.
3
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a
Bitton, G., Masterson, N., Giftord, G.E. (1976). Effect of a Second-
ary Treated Effluent on the Movement of Viruses Through a Cy-
press Dome Soil. J ENVIRON QUAL, 5(4):370-5.
The adsorption of coliphage T2 and poliovirus 1 onto sandy
cyprus dome soil in columns was substantially lower in soil treat-
ed with secondary effluent than in soil treated with tap water. The
secondary effluent also could desorb viruses from soil particles.
It appears that organic materials in waste water effluents may
interfere with the interaction between viruses and soil particles.
Ij
Bitton, G., Pancorbo, 0.., Gifford, G.E. (1976). Factors Affecting
the Adsorption of Polio Virus to Magnetite in Water and Waste-
water. WATER RES, 1O(11):973-80.
The adsorption of poliovirus 1 to magnetite was enhanced by
cations and conformed to the Freundlich isotherm.
At pH levels of 5 to 9, greater than 99% of the virus adsorbed
to the magnetite when the concentration of magnetite exceeded
300 mg/liter and the salt concentrations were 60 mg/liter or
greater.
Bondarenko, V.1., Grigoryeva, L.V. (1975). Isolation of Entero-
viruses from the Surfaces of Vegetables and Fruit. MICROBIOL
ZH, 37(2):244-7. Ukrainian.
Enteroviruses were recovered from the surfaces of vegetables
and fruits grown in fields irrigated with sewage.
Echoviruses 7, 8, 11, and 19, coxsackieviruses A9, B3, and B4
and several unidentified viruses were recovered.
Eighteen percent of 82 carrots and 12°/a of 67 potatoes tested
yielded viruses. Lower recoveries were obtained in tomatoes, rad-
ishes, fruits, berries, lettuce, and cucumbers. Viruses were not re-
covered from onions, parsley, cabbage, or dill suggesting possible
antiviral activity of these vegetables.
The viruses were recovered from anion exchange resin con-
centrated tap water washings of the surfaces of the fruits and
vegetables.
Bouwer, H. (1976). Use of the Earth’s Crust for Treatment or Stor-
age of Sewage Effluent and Other Waste Fluids. CRIT REV ENVI-
RON CONTR, 6(2):111-30.
In a review of the treatment of sewage and the disposal of ef-
fluent on land and by deep well injection, the movement and sur-
vival of viruses in soil and the problems associated with spreading
and spraying effluents on the land are discussed.
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E
Brown, ER., Dolowy, W.C., Sinclair, T., Keith, L., Greenberg, S.,
Hazdra, J.J., Beamer, P., Callaghan, 0. (1976). Enhancement of
Lymphosarcoma Transmission in Esox lucius and its Epidemio-
logic Relationship to Pollution. BIBL HAEMATOL, (43):245-51.
The Mulcahy lymphosarcoma was transmitted by an apparent
cell-free homogenate giving credence to a viral etiology for this
tumor. However, Millipore-filtered material failed to produce tu-
mors in young fish. Fish size, the site of inoculation, temperature
of the water, and age of fish recipients affected the rapidity of
growth of tumor transplants.
Epidemiologically, more and more evidence links organic
chemical pollution to the frequency of tumors. That human enteric
viruses have been detected in fish indicates the seriousness of
these environmental problems.
p.
Buras, N. (1976). Concentration of Enteric Viruses in Wastewater
and Effluent: A Two Year Survey. WATER RES, 1O(4):295-8.
Enteric viruses were recovered throughout the year during a
two-year survey of the wastewater and effluent of the Haifa (Is-
rael) sewage treatment plant. The numbers of enteric viruses in
the wastewater and effluent fluctuated with the seasons and were
usually lowest during the winter. The greatest numbers of viruses
were recovered between the months of April and September. A
peak of 106 x 10 PFU/100 ml occurred in wastewater during one
July, and a peak of 94 x 10 PFU/100 ml occurred in effluent dur-
ing one August.
Carison, S. (1974). Earlier Detection of Virus Epidemics Through
Sewage Investigations. BUNDESGESUNDHBL, 17(25-26):403-5 .
German.
Viruses are present in all sewage effluents. Most of the viruses
found are the same as those circulating in the population and ex-
creted with the feces. From the distribution and concentration of
different virus types in sewage, epidemiological inferences may be
drawn.
y
Carison, S., Hässelbarth, U., Sohn, F.W. (1976). Studies on Virus
Inactivation by Chlorine During Water Disinfection. ZBL BAKT
HYG, 162(3-4)320-9. German.
The ability of free chlorine (HOCI and/or OC ) to inactivate
viruses is better indicated by the oxidation-reduction potential
(ORP) of the free chlorine than by its concentration. A greater
ORP and a longer period of contact is needed to inactivate viruses
than to inactivate bacteria.
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To insure inactivation of polioviruses in water that contains
organic material, an ORP of +750 mV (0.3-0.6 mg/liter of free
chlorine) must be maintained for 15 to 30 minutes.
Adenoviruses and polioviruses are almost equally resistant to
inactivation by chlorine.
Carison, S., H sseIbarth, U.., Sohn, F.W. (1976). Virus Inactivation
by Chlorine—Literature Review. ZBL BAKT HYG, 248:193-215.
German.
The inactivation of viruses by chlorine is extensively reviewed.
Caroli G., Avio, CM. (1975). Isolation of Mycobacteriophages
from Surface Water. ANN SCLAVO, 17(4):568-70.
Mycobacteriophages were recovered from 5 of 15 water sam-
ples taken from canals surrounding the town of Pisa. Eleven of the
samples contained atypical mycobacteria.
CllHe G.G. Current Technology and Advances in True Re-Use of
Effluents. In “Radiation for a Clean Environment.” Proceedings of
the International Symposium on the Use of High-Level Radiation
In Wastewater Treatment—Status and Prospects, Munich, March
1975. InternatIonal Atomic Energy Agency, Vienna, Austria (1975),
29-43.
The removal of viruses from sewage by treatment processes is
briefly discussed within a review of renovation and reuse of
sewage.
rp
Cliver, D.O. (1975). Virus Association with Wastewater Solids.
ENVIRON LETTERS, 1O(3):21 5-23.
Reoviruses and at least five different enteroviruses were re-
covered from the solids of an urban wastewater treatment plant.
Viruses were recovered from sludge digested anaerobically at 30
to 32 C and from grit.
Until reliable means for inactivating viruses in sludge and grit
have been developed and implemented, great care must be taken
in disposing of these solids.
p
Cliver, D.O. Surface Application of Municipal Sludges. In “Virus
Aspects of Applying Municipal Waste to Land,” Symposium Pro-
ceedings, edited by LB. Baldwin, J.M. Davidson, and J.F. Gerber,
Center for Environmental Programs, University of Florida, Gain-
esville (1976), 77-81.
Viruses in wastewater tend to sediment with the sludges that
are formed in treatment. Sludges that contain viable viruses
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should be disposed to soil in a way that precludes run-off. Once
sludges have been properly disposed of at a properly selected
site, the viruses present in the sludges probably represent relative-
ly little risk to public health.
Given the possibility of human error, however, a community
that cannot exert direct control over the surface application of its
sludges is probably well-advised to modify its processes to insure
that all viruses are inactivated before the sludges leave the treat-
ment plants.
iT
Couch, J.A. (1976). Attempts to increase Baculovirus Prevalence
in Shrimp by Chemical Exposure. PROG EXP TUMOR RES,
20:304-14.
In laboratory studies, pink shrimp (Penaeus duorarum)
harvested from areas with various enzootic levels of natural
Baculovirus infection were exposed to low levels of Aeroclor 1254,
a polychlorinated biphenyl (PCB), Mirex, cadmium, and
methoxychior.
No consistent pattern of increase in prevalence of viruses and
no indication of tumor induction was detected.
IT
Couch, J.A., Summers, M.D., Courtney, L. Environmental Signif i-
carice of Bacufovirus Infections in Estuaririe and Marine Shrimp.
In “Pathobiology of Invertebrate Vectors of Disease.” ANN NY
ACAD SCI, 266, New York Academy of Sciences, New York
(1975), 528-36.
A Baculovirus enzootic in populations of penaeid shrimps in
the northern Gulf of Mexico has been discovered. The rod-shaped
virus, both free and occluded, was detected in polyhedral inclu-
sion bodies in the nuclei of host hepatopancreatic cells. The virus
produced striking cytopathologic effects, but induced no gross
signs.
Latent infections have been detected in from 0 to 50% of the
samplings of pink shrimps taken since 1971. The virus has not yet
been grown in established insect cell lines.
The virus was recovered from shrimps taken from waters of
Apalachee Bay, Port St. Joe, and Pensacola, all in Florida. In the
laboratory, the incidence of Baculovirus infections in shrimps was
increased by maintaining the shrimps in large numbers in small
aquariums.
The effect that the Baculovirus has on populations of shrimps
in nature, whether pollutant chemicals in coastal waters affect the
virus in shrimps, and whether the occurrence of this Baculovirus
in a crustacean relates to the development of insect Baculovirus
biopesticides are matters currently under investigation.
7
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y
Cramer, W.N., Kawata, K , Krus , C.W. (1976). Chlorination and
Iodinatson of Poliovirus and f2. J AMER WATER WORKS ASSOC,
48(1):61 -76.
Poliovirus 3 (Leon) was slightly more sensitive to HOCI and
OC and considerably more sensitive to chioramines, particularly
dichloramine, than coliphage f2.
Under dynamic test conditions, in which the disinfectant was
mixed rapidly into the sterilized sewage that contained the virus,
ooliphage f2 and poliovirus 3 were equally and highly sensitive to
iodine at pH levels of 6 and 10. At pH 4, both viruses, especially
the cot Dhage, were destroyed more slowly.
In prereacted studies, in which the disinfectant was mixed
with the sterilized sewage before the virus was added, both viruses
ee rapidly inactivated by iodine at pH 6 (when an iodine resid-
ual persisted), but were unaffected at pH 10 (when an iodine
residual was not detected).
In this study, coliphage f2 was equally or more resistant than
poliovirus 3 to chlorine and iodine.
At pH levels and KI concentrations that favored iodination of
tyrosine and histidine, both viruses were inactivated rapidly mdi-
cating that the mode of action of iodine was similar with both
viruses. At high hydrogen ion concentrations, where the iodination
of tyrosine and histidine is inhibited, both viruses were inactivated
slowly by iodine.
0
Craun, G.F., McCabe, L.J., Hughes, J.M. (1976). Waterborne Dis-
ease Outbreaks in the US—-1971-1974. J AMER WATER WORKS
ASSN, 68(8):4106-10.
Thirteen outbreaks of waterborne hepatitis A, in which 351
people became ill, occurred from 1971 to 1974.
Epidemiologically, hepatitis A has been implicated in 66 out-
breaks of waterborne disease since 1946. Twenty-two of these
outbreaks involved municipal systems, three with inadequate or
interrupted disinfection, five that used contaminated, untreated
surface or ground waters, and eleven that were contaminated by
cross connections or back-siphonage.
Forty-six outbreaks of gastroenteritis of unknown etiology
also occurred from 1971 to 1974. Almost 8,000 cases of illness
resulted.
Waterborne giardiasis, shigellosis, salmonellosis, typhoid fever,
and chemical poisonings that occurred from 1971 to 1974 are also
noted.
Ii.
Darteveile, Z., Desmet, L. (1975). Shigella Research in the Sea and
in an Estuary. Part I. Frequency of Bacteriophages in Polluted
Water. ANN MICROBIOL (Paris), 126B(1):95-7. French.
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This paper was listed in the 1975 edition of these abstracts. A
translation was not then available.
Shigella phages and coliphages were recovered from sewage-
polluted waters of the Gulf of Trieste.
Derbyshire, J.B. (1976). Fate of Animal Viruses in Effluent from
Liquid Farm Wastes. J MILK FOOD TECHN, 39(3):214-7.
Seventeen of 22 samples of liquid manure from a swine fat-
tening house y eIded enteroviruses, adenoviruses, and cor-
onaviruses. An enterovirus was recovered from one of six samples
of waste from a swine farrowing house, but no viruses were recov-
ered from 18 samples of liquid cattle manure from a dairy farm.
Swine enteroviruses were recovered from surface soil samples
collected up to eight days after liquid manure had been spread on
agricultural land. Swine enteroviruses were recovered also from 2
of 26 samples of surface run-off collected from sites at which l q-
uid pig manure had been spread routinely on agricultural land. A
swine enterovirus was recovered from one of 33 samples of sur-
face water collected in areas in which liquid pig manure had been
spread routinely on farm land. No viruses were recovered from
any of 36 samples of ground water collected from such areas.
Field and laboratory experiments indicate that enteroviruses
are more rapidly inactivated in aerated liquid manure than in un-
treated manure.
Dienstag, J.L., Gust, ID.., Lucas, C.R., Wang, D., Purcel%, R.H.
(1976). Mussel-A ssociated Viral Hepatitis, Type-A: Serological
Confirmation. LANCET, 1(7959):561-4.
An epidemiological investigation incriminated incompletely
cooked mussels (Mytilus edulis) as the vehicle of hepatitis A
transmission in seven members of a family of 14 who developed
hepatitis one month after a family outing.
The etiological role of hepatitis A virus was serologically con-
firmed by rises in serum-antibody titers to hepatitis A antigen in
immune adherence hemagglutination tests.
1 .
Dor, I., Schechter, I-I, Shuval, HI. (1976). Biological and Chemical
Succession in Nahal Soreq: A Free-Flowing Wastewater Stream. J
APPL ECOL, 13(2):475-89.
About half of the 21,000 m 3 of undiluted raw sewage that en-
ters the Nahal Soreq (a free-flowing wastewater stream) in Israel
daily is lost by seepage (82%) and evaporation (18°/o). The remain-
der of the sewage undergoes progressive purification in the river
bed.
In a recent study during the dry season, the concentration of
enteric bacteria was reduced by 9 9 % and the concentration of en-
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teric viruses by 95% after 45 km of river flow at a flow rate ot
about 1 km/hour. Concurrently, the BOD was reduced by 97%, the
COD by 90%, and the total suspended solids by 91%.
The purifying capability of the river did not diminish during
the winter except during storms.
a
Dubolse, SM., Moore, B.E,, Sagik, B.P. Surface Application of Mu-
nicipal Effluents. In “Virus Aspects of Applying Municipal Waste to
Land,” Symposium Proceedings, edited by L.B. Baldwin, J.M.
Davidson, and J.F. Gerber. Center for Environmental Programs,
University of Florida, Gainesville (1976), 69-76.
In laboratory studies, poliovirus 2 (Chat) moved for consid-
erable distances through non-sterile soil core samples after peri-
ods of simulated rainfall. In these studies, periods of simulated
rainfall were alternated with periods of application of de-
chlorinated activated sludge effluents.
a
Duboise, S.M., Moore, B.E., Sagik, B.P. (1976). Poliovirus Survival
and Movement in a Sandy Forest Soil. APPL ENVIRON MICRO-
BIOL, 31(4):536-43.
Applications of water and activated sludge effluent (to simu-
late cycles of rainfall and effluent application) onto nonsterile
sandy forest soil produced ionic gradients that affected movement
through the soil of poliovirus 1 that had been seeded into the ef-
fluent. These studies indicated that poliovirus in effluents applied
to soil moved considerable distances through this soil after
rainf aIls.
The capability of the virus to migrate through the soil at 4 C
and 20 C remained unchanged for at least 84 days.
DuPont, H.L. “Culture-Negative” Acute Diarrhea. In “Gastro-Intes-
t lnal Emergencies,” edited by H.R. Clearfield and V.P. Dinoso,
Jr. Grune and Stratton, New York (1976), 269-74.
The association of rotaviruses with more than half of the in-
fantile gastroenteritis that occurs worldwide is noted within a con-
cise review of the microbial etiology, epidemiology, and clinical
concerns in diarrheal disease.
eo o
Earampamoorthy, S., Koff, R.S. (1975). Health Hazards of Bivalve-
Mollusk Ingestion. ANN INTERN MED, 83(1):107-10.
Bivalve mollusks (oysters, clams, and mussels) filter large
quantities of water unselectively and thereby concentrate a variety
of aquatic contaminants pathogenic for man within edible shellfish
viscera.
10
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The recognized bacterial diseases associated with ingestion of
contaminated bivalves include typhoid fever, cholera, and Vibrio
parahemolyticus gastroenteritis. The major known shellfish-asso-
ciated viral diseases are hepatitis A and possibly viral
gastroenteritis.
The ingestion of bivalves that have fed on the toxic dino-
flagellates that produce red tides is responsible for paralytic shell-
fish poisoning. But, outbreaks of airborne respiratory irritation in
populations exposed to red tides may be the most common public
health problem associated with red tides.
The hazards to human health that result from exposure of bi-
valves to industrial, agricultural, and oil pollution in coastal waters,
and the hazards from improper handling of bacterially-con-
taminated mollusks remain to be defined.
V
Engelbrecht, R.S., Amirhor, P. Disposal of Municipal Solid Waste
by Sanitary Landfill. In “Virus Aspects of Applying Municipal
Waste to Land,” Symposium Proceedings, edited by L.B. Baldwin,
J.M. Davidson, and J.F. Gerber. Center for Environmental Pro-
grams, University of Florida, Gainesville (1976), 59-67.
Disposable diapers, other soiled hygiene products, dog and
cat litter, and household products that contact human body open-
ings are sources of viruses in municipal solid wastes. Viruses may
be recovered from landfills and leachates.
Viruses adsorb to solids and other substances in landfills and
are destroyed in leachates by heat (temperatures may reach 55 to
60 C for short periods), generated by aerobic digestion in young
landfills when moisture content is sufficient (40 to 60%), and by
toxic substances that occur in leachates in aging landfills (75 to
215 days).
Some evidence exists which suggests that low molecular
weight fatty acids and heavy metals (iron and zinc) may be re-
sponsible for some leachate toxicity.
Y IP
Epp, C. Experience with a Pilot Plant for the Irradiation of Sewage
Sludge. Experiments on the Inactivation of Viruses in Sewage
Sludge After Radiation Treatment. In “Radiation for a Clean Envi-
ronment.” Proceedings of the International Symposium on the Use
of High-Level Radiation in Wastewater Treatment—Status and
Prospects, Munich, March 1975. International Atomic Energy
Agency, Vienna, Austria (1975), 485-90.
Viruses were detected in eight of 33 samples of sewage
sludge. After exposure to 300 krad from a 6oco source, viruses
were recovered from only two of the 33 samples. Approximately
similar recoveries were attained with poliovirus 1-seeded sludge.
When poliovirus 1 was irradiated similarly in ampules sus-
pended in sludge, about 90% of the virus was inactivated.
11
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3 /p
Epp C. Viro/ogical Studies on the Ozone Apparatus Manufactured
by the Demag Company at the Gross/a ppan Experimental Sewage
Treatment Facility. In “Munich’s Contributions to Sewage, Fishery
and Stream Biology, Vol. 26, Water br Land Reclamation Areas.”
Oldenbourg Verlag, Munich, W. Germany (1975), 191-4. German.
Fourteen grams of ozone/rn 3 of activated sludge effluent de-
stroyed seeded poliovirus in 12 minutes. Eight to nine grams of
ozone/rn 3 of activated sludge effluent did not destroy all of the po-
liovirus in 24 minutes.
a
Fannin, K.F., Spendlove, J.C., Cochran, K.W., Gannon, J.J. (1976).
Airborne Coliphages from Was tewater Treatment Facilities. APPL
ENVIRON MICROBIOL, 31(5):705-10.
The mean levels of coliphages in the airborne emissions of
trickling-filter beds and activated sludge treatment plants (that
contained phage) were 2.84 x 10’ and 3.02 x 10.1 MPN/m 3 , respec-
tively. The coliphage levels in the sewage were about 5 x 10 to
3 x 106 PFU/liter.
Wind velocity and temperature did not affect the airborne con-
centration of coliphages, but airborne coliphage concentrations
increased as humidity increased.
Darkness may have also favored survival of airborne
coliphages.
All air samples were taken 2 to 15 meters from the edge of the
plants at a height of 1 .5 meters.
27
Parley, CA. (1976). Ultrastructural Observations on Ep zootic Neo-
p/asia and Lytic Virus Infection in Bivalve Mollusks. PROG EXP
TUMOR RES, 20:283-94.
Epizootic neoplasia occurs in marine bivalve mollusks.
Epidemic sarcomas occurred in oysters and mussels from
Oregon in incidences of 12 to 40% in the fall, winter, and spring.
A recent epizootic in clams (Macoma balthica) from Maryland
exhibited the same epizootiologic seasonality and occurred in a
localized geographic area. Lesions developed in the gills and in-
vaded tissues throughout the animal. Ultrastructural evidence indi-
cated epithelial origin of the neoplastic cells. Degenerative and
preneoplastic changes in the epithelium were also evident.
A virus morphologically similar to papillomavirus has been
discovered that replicates in germ cells of oysters (Crassostrea
virginica).
3 /
Farooq, S., Chian, E.S.K. (1976). Discussion—/nactivation of Polio-
virus in Water by Ozonat,on, J WATER POLLUT CONTRL FED,
48(3) :593-8.
12
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This paper is a discussion of a study by Majumdar, Ceckler,
and Sproul (Majumdar, S.B.., Ceckler, W.H., Sproul, O.J. (1973).
inactivation of Poliovirus in Water by Ozonation, J WATER POL-
LUT CONTRL FED, 45(12):2433-43; Majumdar, S.B., Ceckler,
W.H., Sproul O.J. (1974). Communication—Inactivation of Polio-
virus in Water by Ozonation, J WATER POLLUT CONTRL FED,
46(8):2048-53) in which the data on the kinetics of the inactivation
of poliovirus 1 by ozone are reanalyzed.
See 1973 and 1974 Literature Abstracts.
A
Farrah, S.R., Gerba, C.P., Wallis, C., Melnick, J.L. (1976).
Concentration of Viruses from Large Volumes of Tap Water Using
Pleated Membrane Filters. APPL ENVIRON MICROBIOL,
31(2):221 -6.
Poliovirus 1 was recovered from seeded tap water (pH adjust-
ed to 3.5 and AICI 3 to 0.0005 M final) by adsorbing the virus onto
a 10-inch (25.4 cm) fiberglass depth cartridge and a 10-inch
pleated epoxy-fiberglass filter arranged in series in a Wallis-Mel-
nick concentrator. Flow rates up to 37.8 liters/minute were
achieved.
Adsorbed viruses were eluted from the filters with glycine
buffer (pH 10.5), the eluate was reconcentrated by an aluminum
flocculation process, and the viruses were eluted from the alumi-
num floc with glycine buffer (pH 11.5).
With this procedure, an average of 40 to 50% of the viruses in
1,900 liters (500 gallons) of tap water were recovered in a period
of three hours.
A
Farrah, S.R., Goyal, S.M., Gerba, C.P., Wallis, C., Shatter, P.T.B.
(1976). Characteristics of Humic Acid and Organic Compounds
Concentrated from Tapwater Using the Aquella Virus Concen-
trator. WATER RES, 1O(1O):897-901.
Humic acids and other organic compounds in eluates from fil-
ers used in processing large volumes of tap water with the Wallis-
Melnick virus concentrator adsorbed to membrane filters at low
pH, eluted from membrane filters at high pH, interfered with virus
adsorption to membrane filters, and formed flocs at low pH. The
hurnic acids were removed from the eluates by anion-exchange
resins.
A
Farrah, S.., Wallis, C., Shatter, P.T.B., Melnick, J.L. (1976).
Reconcentration of Poliovirus from Sewage. APPL ENVIRON
MICROBIOL, 32(5):653-8.
Certain soluble components in sewage adsorb on virus ad-
sorbent filters and are removed along with the viruses by elutants.
Upon acidification of eluates to reconcentrate the viruses on
13
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smaller membrane surfaces, flocs form that interfere with the
reconcentration process. The interfering sewage components may
be removed from eluates by activated carbon and ion-exchange
resins.
A
Fatlal, B., Katzenelson, E. (1976). Evaluation of Gauze Pad Method
to Recover Viruses from Water. WATER RES, 1O(12):1 135-40.
In field studies, the gauze pad method was more effective for
detecting enteroviruses in a sewage stream than two-liter volume
grab samples concentrated by the phase separation method.
In laboratory experiments, the recovery efficiency of the
gauze pad method for concentrating viruses from tap water was
very low and depended upon sample volume—the larger the vol-
ume the lower the efficiency.
y
Floyd, R., Johnson, JO., Sharp, 0G. (1976). Inactivation by Bro-
mine of Single Poliovirus Particles in Water. APPL ENVIRON
M1CROBIOL, 31(2):298-303.
Quantitative electron microscopy showed that in Freon-ex-
tracted poliovirus 1 preparations velocity banded in a sucrose
gradient, more than 95% of the virions were present as single
particles.
At 2 C, these well-dispersed virions were inactivated in rapidly
flowing 2.2 M bromine at pH 7 at the rate of one log 10 unit/lU
seconds.
The rate of inactivation of the virions increased with in-
creasing temperature. At 10 C and 30 C, the rate of inactivation in-
creased approximately linearly with increasing bromine concen-
tration. At 2 C, a linear relationship did not occur.
Fox, J,P. Human-Associated Viruses in Water. In “Viruses in Wa-
ter,” edited by G. Berg, H.L. Bodily, EH. Lennette, J.L. Melnick,
T.G. Metcalf. American Public Health Association, Washington,
D.C. (1976) 39-49.
Contaminated water may be a vehicle for transmitting viruses
to man when the water is used for drinking, washing eating uten-
sils, dishes, and food to be consumed without cooking, brushing
teeth, bathing, swimming, and irrigation of vegetables. Shellfish
harvested from sewage-polluted waters may also be a vehicle for
transmitting viruses to man.
Feces and urine are the most important sources of human
viruses in waters. Enteroviruses and adenoviruses, hepatitis A vi-
rus, and the viruses of acute infectious non-bacterial gastro-
enteritis are among those most likely to be transmitted by water.
In developed countries, transmission of these agents through
drinking water supplies is likely only if breaks occur in the treat-
14
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ment procedures. In undeveloped countries, waterborne spread is
presumedly extensive and may constitute safe natural immuniza-
tion when it occurs early in life.
Freytag, B. Hygienic Aspects of Swimming in Lakes. In “Munich’s
Contributions to Sewage, Fishery and Stream Biology, Vol. 26,
Water for Land Reclamation Areas.” Oldenbourg Verlag, Munich,
West Germany (1975), 111-12. German.
The significance of viruses in bathing waters is evaluated
within a broad discussion of the sanitation of swimming lakes.
U
Gilbert, R.G., Gerba, C.P., Rice, R.C., Bouwer, H., Wallis C., Mel-
nick J.L . (1976). Virus and Bacter,a Removal from Wastewater by
Land Treatment. APPL ENVIRON MICROBIOL, 32(3):333-8.
Neither viruses nor Salmonella were detected in renovated
waters from four wells at the Flushing Meadows Wastewater Reno-
vation Project near Phoenix, Arizona. The waters were assayed bi-
monthly in 1974 during flood periods.
Filtration of wastewater through nine meters of soil decreased
the numbers of fecal coliforms, fecal streptococci, and total bac-
teria in the renovated well waters by about 99.9%.
0
Gilbert, R.G., Rice, R.C., Bouwer, H., Gerba, C.P., Wallis, C., Mel-
nick, J.L. (1976). Wastewater Renovation and Reuse: Virus Re-
moval by Soil Filtration. SCIENCE, 192(6):1004-5.
Effluent from an activated sludge sewage treatment plant at
the Flushing Meadows Wastewater Renovation Project near Phoe-
nix, Arizona, sampled bimonthly, yielded an average of 2,118 virus
infective units/100 liters (range 158 to 7,475 infective units/lOU
liters).
Viruses were not detected in wells at depths of 6 to 9 meters
in sand infiltration basins during the second seven days of 14-day
effluent charge periods. The sand appeared to remove at least
99.99% of the viruses from the effluent.
The viruses were received by membrane adsorption-elution vi-
rus concentrator technics.
Globa, L.I., Rotmistrov, M.N., Bember, V.E., Grebenyuk, V.D.,
Gvozdyak, P.1 (1975). Freeing Water from Viruses by Electro Dia-
lysis. ELEKTRON OBRAB NATER, 4:57-9. Translation presently
not available. Russian.
15
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Goldfleld, M. Epidemiological Indicators for Transmission of
Viruses by Water. in “Viruses in Water,” edited by G. Berg, H.L.
Bodily, E.H. Lennette, JL. Melnick, T.G. Metcalf. American Public
Health Association, Washington, D .C. (1976), 70-85.
Ample evrdence indicates that too many people in the United
States too often are provided with unsafe water supplies. Although
potable public water supplies may be treated in inadequate facili-
ties operated by inadequately trained personnel, dispensed
through faulty distribution systems and woefully monitored as to
water quality, the situation with respect to semi-public water sup-
plies is even worse.
Generally, the responsibility for inspecting semi-public sup-
plies, where they are inspected with any regularity or frequency
at all. is left to locally employed sanitarians who may be inade-
quately trained, poorly motivated, virtually unsupervised and sub-
ject to strong local political pressures to overlook defects. In the
United States, most semi-public supplies are unchlorinated and
when tested at all, are frequently tested in laboratories of inade-
quate competence.
The general public, and even large segments of the medical
community—have a grossly distorted impression of surveillance of
disease in the United States. Over the years, they have been led to
believe that some system exists for recognizing disease outbreaks
that provides reasonable assurance that significant clusters of ill-
ness will be recognized, studied, and handled appropriately by
public health authorities. To them, the absence of reported out-
breaks provides reassuring evidence of a safe and healthful envi-
ronment. Unfortunately, this is pure myth. Even huge waves of
febril respiratory disease, gastrointestinal illness, measles, or ger-
man measles can go completely undetected unless arduous efforts
are made to develop surveillance programs. In New Jersey. out-
breaks of a disease as dramatic in its consequences as eastern
encephalitis went completely undetected until a study was mounted
to search for its occurrence.
Hepatitis A virus is the single virus thus far associated with
waterborne outbreaks, probably because it frequently produces
overt disease in susceptible populations, and because the overt
disease produced is relatively easily recognized as an entity. Other
enteroviruses, however, may eventually be shown to be far more
commonly transmitted by drinking water than hepatitis A virus is.
4:0
Grabow, W.O.K. (1976). Progress in Studies on the Type A (Infec-
tious) Hepatitis Virus in Water. WATER SA, 2(1):20-4.
The nature, epidemiology, clinical significance, and the trans-
mission by water of hepatitis A and B viruses are reviewed.
Data on recent findings which indicate that lesser bushbabies
(Ga/ago senegalensis moho!i) may be susceptible to hepatitis A vi-
rus are presented.
16
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Gude, G. The Legislative Concern for Research on Viruses in
Waste and Potable Waters. in “Viruses in Water,” edited by G.
Berg, H.L. Bodily, E.H. Lennette, J.L. Melnick, T.G. Metcall. Ameri-
can Public Health Association, Washington, D.C. (1976), 27-35.
Against a background of current legislative efforts, the history
of the treatment of sewage, and the control of waterborne infec-
tious diseases throughout the world, Congressman Gilbert Gude
discusses the oncoming needs for water renovaUon and for meth-
ods to assure that renovated waters will be free of viruses.
Li
Guy, M.D., Mclver, J.D. Lewis, M.J. (1976). The Application of the
Foam Fractionation Process to the Removal of Viruses Part I. The
Production of A Mathematical Model to Predict the Efficiency of
Virus Removal. WATER RES, 10(8):737-9.
Removal of coliphage MS2 by Arquad T50 (a cationic surfac-
tant composed of a blend of alkyl quaternary ammonium chlor-
ides) in a laboratory scale foam fractionation plant was brought
about by adsorption of the virus to the surf actant and by the phys-
ical removal of free virus particles entrapped in interstitial liquid.
Guy, M.D., Mclver, J.D., Lewis, M.J. (1976). The Application of the
Foam Fractionation Process to the Removal of Viruses. Part Ill. An
Investigation of the Toxicity of the Surfactant. WATER RES,
10(8): 742-4.
The cationic surfactant Arquad T50 (a blend of alkyl quater-
nary ammonium chlorides) was toxic to coliphage MS2.
An attenuated poliovirus, coxsackievirus B3, and echovirus 11
were relatively unaffected by the surfactant.
Hattingh, W.H.J., Nupen, EM. (1976). Health Aspects of Potable
Water Supplies. WATER SA, 2(1):33-46.
In the first two years of a ten-year research program to assess
the chemical and microbiological quality of potable reclaimed ef-
fluents and other potable waters, the quality of reclaimed waters
was at least as good as that of treated surface waters.
To date, viruses have not been detected in reclaimed water.
11
Hill, BJ. Properties of a Virus isolated from Bi-Valve Mo//usc Tell-
ma tenuis (Da Costa). In “Wildlife Diseases,” edited by L.A. Page.
Plenum Press, New York, New York (1976), 445-52.
A virus was recovered from a marine bivalve molkisc (Tellina
tenuis) with a disease of the digestive gland. Morphologically, the
virus resembled the fish pathogen infectious pancreatic necrosis
(IPN) virus. The two viruses did not cross react in neutralization
17
-------
tests, but an antigenic relationship between the two viruses was
demonstrated in fluorescent antibody tests.
The newly recovered virus contained RNA in its nucleic acid,
had a growth temperature optimum near 15 C, was resistant to
lipid solvents, was stable at pH 3, at -20 C in 50% glycerol, and at
-20 C and -70 C in the frozen state.
The virus infected Tel/ma terluis and also the Japanese oyster
(Crassostrea gigas).
A
Hill, W.F.., Jr., Jakubowski, W., Akin, E.W., Clarke, NA. (1976).
Detection of Virus in Water. Sensitivity of the Tentative Standard
Method for Drinking Water. APPL ENVIRON MICROBIOL,
31(2):254-61.
The sensitivities of nitrocellulose membrane filters, epoxy-
fiberglass-asbestos filters, yarn-wound fiber glass depth filters, and
epoxy-fiber glass filter tubes for detecting small quantities of
viruses in large quantities of drinking water were investigated. The
test procedure was that proposed as a tentative standard method
in the fourteenth edition of Standard Methods for the Examination
of Water and Wastewater (APHA, AWWA, WPCF, Washington,
D.C.. 1976). Viruses were adsorbed to the filter media at pH 3.5
and eluted with glycine buffer, pH 11.5.
In 120 tests with poliovirus 1, the efficiencies of virus recov-
eries ranged from 28 to 42°/o; the average efficiency was 35%. In 44
tests with virus input levels of 15 to 35 PFU/1 900 liters of water
sampled, the poliovirus was detected 95% of the time. In 76 tests
with virus input levels of 10 PFU or less/i ,900 liters, the virus was
detected 66% of the time.
No significant difference was observed in the virus detection
sensitivities of the various virus-adsorbent media tested.
At inputs of 36 to 100 PFU or TCID 50 /volume of water tested.
coxsackieviruses A21 and B6, echoviruses 1, 13, 24, and 27 and
adenovirus 15 were detected in 380- and 1,900-liter quantities of
water.
y
Hoehn, R.C. (1976). Comparative Disinfection Methods. J AMER
WATER WORKS ASSN, 68(6):302-8.
The problems of viruses in water are briefly discussed within a
review on methods for disinfecting water.
Hughes, D.E., Stafford, D.A. (1976). The Microbiology of the Acti-
vated-Sludge Process. CRIT REV ENVIRON CONTRL, 6(3):233-56.
The problems of viruses in sewage are discussed briefly with-
in an extensive analysis of the microbiology of the activated
sludge process.
18
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iT
Johnson, PT. (1976). A Herpes/ike Virus from the Blue Crab, Cal-
linectes sapidus. J INVERT PATHOL, 27(3):419-20.
Herpesvirus-like particles were detected by electron-microsco-
py in hematopoietic tissue of a Blue Crab (Callinectes sapidus)
harvested from Chincoteague Bay. The particles were hexagonal
and pentagonal, approximately 118 nm in diameter, and contained
a nucleoid. Larger particles were seen that may have been en-
veloped. The nuclear material of the cells that contained the virus-
like particles was Feulgen-positive suggesting that the particles
contained DNA.
This is the second report of a herpesvirus-like particle in in-
vertebrates; a herpesvirus-like particle was detected earlier in
heat-stressed oysters (Crassostrea virgin/ca).
Kalitina, T.A. (1975). A Method for Isolating Entero viruses in Meat.
VOPR PITAN, (6):52-5. Russian.
Echovirus 11 was detected in stewed beef that had been seed-
ed with 200 ID 50 of the virus/gm, but not in beef that had been
seeded with 20 1D 50 /gm. The virus was also detected in meat cut-
lets that had been seeded with 3 x 10 1D 50 /gm, but not in cutlets
seeded with 30 1D 53 /gm.
All of the poliovirus was recovered from beef seeded with
more than 106 lD by elution of the virus with 8 to 9 volumes of
saline.
Kalter, S.S., Mitistein, C.H. Animal-Associated Viruses in Water. In
“Viruses in Water,” edited by 0. Berg, H.L. Bodily, E.H. Lennette,
J.L Melnick, 1.0. Metcalf. American Public Health Association,
Washington, D.C. (1976), 50-60.
Viruses of many different types and heterogeneous non-hu-
man sources may find their way into water supplies. These include
viruses of farm animals and wild life, fish, amphibia, plants, in-
sects, and bacteria. Although few, if any, of these viruses produce
acute diseases in man, it is not known whether any of these
viruses produce latent infections or tumors in man.
8
Kantoch, M., Dobrowo lska, H., Jarzabek, Z., Jankowski, M. (1975).
Studies in Distribution and Characteristics of Poliovirus Strains
Circulating in Poland in 1966-1972. MED DOSW MIKROBIOL,
27(4):365-72. Polish.
Epidemic poliovirus strains recovered in Poland in 1968, es-
pecially early in the year, were antigenically related to USOL-D-
bac. All of the strains tested were distant antigenically from Leon
12a 1 b.
19
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Strains from sporadic cases and from sewage contained rct
and 1ST markers that were close to, intermediate, and distant from
those typical of the attenuated strains. From sporadic cases in
1970 to 1972, strains with antigenic structure similar to that of the
vaccine attenuated strains were recovered.
A
Katzenelson, E. Virologic and Engineering Problems in Monitoring
Viruses in Water. In “Viruses in Water,” edited by G. Berg, H.L.
Bodily, E.H. Lennette, J.L. Melnick, T.G. Metcalf. American Public
Health Association, Washington, D.C. (1976), 152-64.
Our capability for recovering small numbers of viruses from
large volumes of water is limited by factors peculiar to viruses and
by shortcomings of viral concentration and recovery technics , The
many viruses that may occur in water cannot all be detected in the
same cell culture and animal systems and some cannot yet be re-
covered in any. Neither are all viruses recovered with equal effi-
ciency by concentration methods. The effectiveness of concen-
tration methods is affected by the quality of water under test.
Except with new technics such as fluorescent antibody meth-
ods currently under test, days to weeks are required for recov-
ering and identifying viruses in various waters.
‘ I
Katzenelson, E., Bledermann, N. (1976). Disinfection of Viruses in
Sewage by Ozone. WATER RES, 1OfT):629-31.
When poliovirus 1-seeded filtered sewage was added to buff-
ered ozone, the ozone disappeared immediately. The concen-
tration of virus was reduced immediately, but the disappearance of
ozone was accompanied by a stabilization of virus titer in the ef-
fluent. The extent of virus inactivation depended upon the concen-
tration of ozone and on the amount of interfering organic matter
in the effluent.
When ozone was bubbled into poliovirus 1-seeded filtered
sewage, inactivation of the virus began after about 30 seconds,
even before ozone was detected in the sewage. Inactivation of
99.999% of the virus occurred in two minutes with an ozone re-
sidual of about 0.6 mg/liter. When ozone was bubbled into virus-
seeded buffer, 99 . 9 9 9 % of the poliovirus was inactivated in less
than one minute with an ozone residual of about 1.0 mg/liter.
6
Katzenelson, E., Bulum, 1, Shuval, Hi. (1976). Risk of Commu-
nicable Disease Infection Associated with Was tewater Irrigation in
Agricultural Settlements. SCIENCE, 194(4268) :944-6.
The incidence of shigellosis, salmonellosis, typhoid fever, and
infectious hepatitis were two to four times higher in 77 kibbutzim
(agricultural communal settlements in Israel) that practiced spray
irrigation with non-disinfected oxidation pond effluent than in 130
kibbutzim that practiced no form of wastewater irrigation.
20
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No significant differences in the incidences of streptococcal
infections, tuberculosis, and laboratory-confirmed cases of influ-
enza were apparent in the two groups. No differences in enteric
disease rates during the winter non-irrigation season were evident
in the two groups either.
A
Katzenelson, E.,, Fattal, B.., Hostovesky, T. (1976). Organic Floccu-
lation: an Efficient Second-Step Concentration Method for the De-
tection of Viruses in Tap Water. APPL ENVIRON M1CROBIOL,
32(4) :638-9.
A method is described for second-step concentration of
viruses from water. This method, combined with an adsorption-
elution method, yields a mean recovery of about 75%.
Poliovirus 1, seeded into 3% beef extract, was recovered with
an average efficiency of 100% by precipitating the beef extract at
pH 3.5 or 4.0 and dissolving the precipitate in a small quantity of
0.15 M Na 2 HPO 4 at pH 9. A 20-fold concentrat;on of the virus was
achieved.
This organic flocculation procedure, applied as a secondary
concentration step after adsorption of poliovirus I from 500 liters
of seeded tap water onto 0.45 .i.m pore size epoxy-fiberglass-asbes-
tos filters (AA Cox M 780), and elution of the adsorbed virus with
300 ml of 3 % beef extract at pH 9.0, resulted in an average virus
recovery of 74.4% (range 60-9l°/o). Parallel studies with the ten-
tative standard method (Standard Methods for the Examination of
Water and Wastewater APHA, AWWA, WPCF, Washington, D.C.,
1976), in which elution of the virus from the membrane is achieved
with glycine buffer at pH 11.5 and reconcentration is brought
about by readsorption on a smaller membrane at pH 3.5 and a
second elution in a smaller volume of elutant, yielded a recovery
of only an average 35% (range 30-47%).
p
Kiker, C.F. Economic Aspects of Virus Inactivation in Waste Treat-
ment. In “Virus Aspects of Applying Municipal Waste to Land,”
Symposium Proceedings, edited by L.B. Baldwin, J.M. Davidson,
and J.F. Gerber. Center for Environmental Programs, University of
Florida, Gainesville (1976), 137-53.
The economics for destroying viruses in sludges is permeated
with uncertainty. Policy makers have little benefit and cost infor-
mation available upon which to base decisions.
Communities that take early actions and build facilities for re-
moving and destroying viruses in sludges and wastewaters will
create necessary decision-making knowledge for those commu-
nities that do so later.
21
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e 1T 60
Krels 1 G.J.., Gassner M., Blum, A.L. (1975). Epidemiology of !rifec-
lious Hepatitis. GASTROENTEROL FORTBIL.DUNGSK PRAXI 5:1-
18. German.
Water- and shellfish-transmitted hepatitis f are briefly dis-
cussed within a general review of infectious hepatitis.
a
Lance, J.C., Gerba, C.P.., Melnick, J.L. (1976). Virus Movement in
Soil Columns Flooded with Secondary Sewage Effluent. APPL EN-
VIRON MICROBIOL, 32(4):520-6.
Secondary sewage effluent seeded with 3 x 1O PFU of polio-
virus I (LSc)/ml was passed through columns 250 cm in length
packed with calcareous sand from an area in the Salt River bed
used for groundwater recharge with secondary effluent. Viruses
were detected in 5 of 43 100-mI samples of the drainage water
from the column. Most of the viruses were adsorbed in the top 5
cm of soiL Viruses were not detected in 1-mt samples extracted
from the columns below the 160 cm level.
Infiltration rates between 15 and 55 cm/day did not affect the
degree of removal of viruses. The top several centimeters of soil
did not saturate with viruses and the movement of viruses was not
affected when the column was saturated continuously for 27 days
with the sewage-virus mixture. Deionized water desorbed viruses
from the soil and increased their movement through the columns.
The addition to the columns of deionized water that contained
CaCI 2 and then sewage started a virus front moving, but the viruses
were readsorbed and none was detected in outflow samples.
Desorption was reduced when soils were dried for one day
between application of viruses and flooding with deionized water.
Drying for 5 days prevented desorption.
Large reductions (99.9 i% or mote) in virus numbers should
occur after passage of secondary effluent through 250 cm of cal-
careous sand similar to that used in the columns unless heavy
ra ns fall within one day after the application of sewage. The
movement of viruses through soils in recharge basins might be
minimized by the proper management of flooding and drying
cycles.
Larkin, E.P.., Tiemey, J.T., Sullivan, R. Persistence of Poliovirus 1
in Soil and on Vegetables Irrigated with Sewage Wastes: Potential
Problems. in “Virus Aspects of Applying Municipal Waste to
Land” Symposium Proceedings, edited by LB. Baldwin, JM.
Davidson, and J.F. Gerber. Center for Environmental Programs,
University of Florida, GaInesville (1976), 119-30.
Poliovirus 1, flooded onto soil in seeded sewage sludge and
effluent, survived for at (east 96 days during the winter months.
22
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On vegetables, spray-irrigated with seeded effluent, the virus sur-
vived for 36 days during the summer.
The application of sewage sludges and effluents onto fields
on which crops that are consumed raw are grown is hazardous.
Contaminated crops that enter the household in the raw state and
later are cooked are also hazardous, because such foods, before
they are cooked, may contaminate other foods.
A
Laveran, H., Beytout, D., Charrier, F. (1976). Adsorption of Entero-
viruses on Cellulose Nitrate Membranes: Influence of Mineral
Salts, Detergents, and Calf Serum. ANN MICROBIOL, 127(3):439-
46. French.
Poliovirus 2 in distilled water, with or without added salts
(NaCI, MgCI 2 , AICI 3 ), adsorbed to cellulose nitrate membranes.
The greatest adsorption occurred when NaCI was present. Ad-
sorption did not occur when detergent was added to the virus sus-
pensions or when membranes were treated with calf serum before
the virus was filtered through.
The adsorbed enteroviruses were eluted from the membranes
with alkaline or proteinaceous buffers.
‘cyp
Lemke, H.S., Sinskey, A.J. Viruses and Ionizing Radiation in Re-
spect to Waste-Water Treatment In “Radiation for a Clean Envi-
ronment.” Proceedings of the Internationi Symposium on the Use
of High-Level Radiation in Wastewater Treatment—Status and
Prospects, Munich, March 1975. International Atomic Energy
Agency, Vienna, Austria (1975), 99-120.
The effects of ionizing radiation on viruses are reviewed. The
radiosensitive target of ionizing radiation is believed to be the nu-
cleic acid of the virus. Temperature and the nature of the sus-
pending menstruum affect the rate of the inactivation reaction.
The effect of oxygen on the reaction is unclear.
Lennette, E.H. Problems Posed to Man by Viruses in Municipal
Wastes. In “Virus Aspects of Applying Municipal Waste to Land,”
Symposium Proceedings, edited by L.B. Baldwin, J.M. Davidson,
and J.F. Gerber. Center for Environmental Programs, University of
Florida, Gainesville (1976), 1-7.
Whether terrestrial deposits of municipal wastes are an infec-
tious disease threat depends upon the pathogens in the wastes
and on factors in the circumambient environment.
All available epidemiological data point to person-to-person
transmission as the principal mechanism of infection with viruses
in excreted wastes. Although viruses may be present in solid and
other wastes discharged to the environment, public health workers
23
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should base recommendations for protecting populations from
hazards in the environment on established ep demiological pat-
terns rather than upon presumptions of possibility.
Lewis, N.F. Scope for Utilizing Gamma Radiation for Micro-
biological Control of Sewage Waste-Water in India. In “Radiation
for a Clean Environment.” Proceedings of the International Sym-
posium on the Use of High-Level Radiation in Wastewater Treat-
ment—Status and Prospects, Munich, March 1975. International
Atomic Energy Agency, Vienna, Austria (1975), 197-202.
The doses of ionizing radiation that are lethal for viruses are
briefly reviewed within a discussion of the usefulness of gamma
radiation for the control of the microbial populations in sewage.
Lo, S., Gilbert, J., Hetrick, F. (1976). Stability of Human Entero-
viruses in Estuarine and Marine Waters. APPL ENVIRON MICRO-
BIOL, 32(2):245-9.
Salinity did not affect the survival of poliovirus 1, echovirus 6,
or coxsackievirus B5 in autoclaved artificial sea water under con-
trolled laboratory conditions or in autoclaved natural sea water. In
the natural sea water system, the viruses and the sea water were
placed in dialysis tubes and suspended in free-flowing estuarine
or sea water. The viruses survived much longer in the sterile arti-
ficial sea water than in the sterile natural sea water.
In the natural sea water system, less than 99% of cox-
sackievirus B5 was destroyed in 80 days at 4 C to 16 C (winter),
although 99.999% of the virus was destroyed in less than 50 days
at 21 C to 26 C (summer). The poliovirus 1 and the echovirus 6
were destroyed more rapidly under similar conditions.
jY
Lund, E. Public Health Aspects of Waste-Water Treatment. In
“Radiation for a Clean Environment.” Proceedings of the Inter-
national Symposium on the Use of High-Level Radiation in Waste-
water Treatment—Status and Prospects, Munich, March 1975. In-
ternational Atomic Energy Agency, Vienna, Austria, (1975), 45-60.
The bacteria, viruses, and protozoan parasites in waste and
polluted waters that are pathogenic to man are described. The ef-
ficiencies of conventional wastewater treatments in removing the
pathogens is reviewed.
At present, conventional treatments cannot produce effluents
free from pathogens that are present in untreated wastewaters.
Pathogens in sludges may be destroyed by pasteurization.
24
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Lund, F. Disposal of Sludges. In “Viruses in Water,” edited by G.
Berg, H.L. Bodily, E.H. Lennette, J.L. Melnick, T.G. Metcalf. Ameri-
can Public Health Association, Washington, D.C. (1976), 196-205.
Viruses were recovered more frequently from primary sludges
than from corresponding raw sewage. The concentration of viruses
in these sludges ranged from 10 to 1,000 TCD 50 !ml. Most of the
viruses in slidges are bound to particulate matter.
Secondary sludges yield only 1 to 10% of the number of
viruses recovered from primary sludges. Sludges from chemical
flocculation of effluents also contain viruses. Even limed sludges
formed at pH 10.5 have yielded viruses.
Viruses have also been recovered from sludges digested at 37
and 50 C. Land disposal of sludges may portend hazards to
health. Efforts should be made with most sensitive methods avail-
able to detect viruses in sludges before those sludges are dis-
posed to the environment.
1
Mackowiak, P.A., Caraway, C.T., Portnoy, B.L. (1976). Oyster-As-
sociated Hepatitis: Lessons from the Louisiana Experience. AMER
J EPIDEMIOL, 103(2):181-91.
An outbreak of oyster-associated hepatitis that occurred in
Louisiana during October and November, 1973 was apparently re-
lated directly to two similar outbreaks that occurred at the same
time in Houston, Texas and Calhoun, Georgia. The oysters incrim-
inated in all three outbreaks were traced to approved oyster grow-
ing areas along the east Louisiana coast. Fecal pollution of the
oyster beds may have been brought about by flooding of the Mis-
sissippi River.
Epidemiologic and environmental data were consistent with
recent experimental evidence which indicated that under natural
conditions shellfish eliminate bacteria and viruses at different
rates and may retain certain enteroviruses for as long as two
months after they have disappeared from surrounding waters.
t O
Mahdy, M.S. Geographic Considerations in Virus Removal in the
Americas. In “Viruses in Water,” edited by G. Berg, Hi. Bodily,
E.H. Lennette, J.L. Melnick, T.G. Metcalf. American Public Health
Association, Washington, D.C. (1976), 221-35.
Geographic factors that affect the problems that are posed by
viruses in the environment may be man-related, virus and virus-
host-related, or environment-related. Man-related factors include
population densities, occupations, socic-econornic conditions, and
sanitation. Virus- and virus-host related factors include differences
in virus types and strains recovered from different areas, recov-
erability of viruses by available technics, quantities and duration
of virus shedding, ratios of inapparent to apparent infection, and
25
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sensitivities of viruses to the environment. Environment-related
factors include areas of inhabitable land, climatic and topographic
conditions, availability and quality of water, and presence of vec-
tors of virus transmission.
An analysis of data on the incidences of hepatitis A in differ-
ent areas of the United States and on the qualities of water sup-
plies, as measured by coliform determinations, suggested a cor-
relation between the incidence of the disease and the quality of
the available water.
Malina, J.F., Jr. Viral Pathogens Inactivation During Treatment of
Municipal Wastewater. In “Virus Aspects of Applying Municipal
Waste to Land,” Symposium Proceedings, edited by L.B. Baldwin,
J.M. Davidson, and J.F. Gerber. Center for Environmental Pro-
grams, University of Florida, Gainesville (1976), 9-23.
The inactivation and removal of viruses from sewage and
sludges by treatment procedures are reviewed.
The lack of parallelism in the inactivation and removal of po-
lioviruses and bacteriophages by these processes is noted.
(0
Maynard, J.E. (1976). Hepatitis A. YALE J BIOL MED . 49(3):227-
33.
The transmission of hepatitis A by water is briefly noted with-
in a broad review of the disease.
Melnick, J.L. Viruses in Water. An Introduction. In “Viruses in Wa-
ter,” edits by G. Berg, H.L. Bodily, E.H. Lennette, J.L. Melnick,
T.G. Metcalf. American Public Health Association, Washington,
D.C. (1976), 3-11.
The author recounts his experiences as an early water virolo-
gist from his persistant recoveries of polioviruses from New York
City sewage during poliomyelitis outbreaks in the mid 1940’s,
through the massive hepatitis A epidemics in New Delhi in 1955,
to the present.
3 ,
Melnick, JL, Gerba, C.P., Wallis, C., Hobbs, M.F. Photodynamic
Inactivation of Virus in Sewage. In “Virus Aspects of Applying Mu-
nicipal Waste to Land,” Symposium Proceedings, edited by L.B.
Baldwin, J.M. Davidson, and J.F. Gerber. Center for Environmental
Programs, University of Florida, Gainesville (1976), 25-36.
Because of the requirements for elevated pH and for light,
which is energy-intensive, photodynamic oxidation is usually a
more expensive disinfection process than chlorination. But, in wa-
ter treatment plants where the pH-elevating lime-softening process
26
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is used, the cost of photodynamic oxidation is greatly reduced.
Moreover, sunlight is an effective alternative energy source to ex-
pensive artificial light.
Disinfection by photodynamic oxidation may be an especially
useful alternative to chlorination in situations where the formation
of chlorinated carcinogenic compounds must be avoided.
Merson, M.H., Morris, G.K., Sack, D.A., Wells, J.G., Feeley, J.C.
Sack, R .B., Creech, W.B., Kapikian, A.Z., Gangarosa, E.J. (1976).
Travelers’ Diarrhea in Mexico. NEW ENGLAND J OF MED,
294(24):1 299-1305.
Forty-nine percent of 73 physicians and 48 family members
who attended a medical congress in Mexico City and participated
in a prospective study of travellers’ diarrhea became ill with diar-
rhea. The median duration of illness was five days. The median
day of onset was six days after arrival.
An etiologic agent was recovered from 6 3 % of the patients.
Enterotox igen Ic Escherichia co/i of different, non-entero-
pathogenic serotypes appeared to be the most common causes of
the diarrheas; other apparent etiologic agents included sal-
monellae, invasive E. co/i, shigellae, Vibrio parahaemolyticus,
Giardia lamb/ia and a reovirus-like agent. Consumption of salads
that contained raw vegetables was associated with enterotoxigenic
E. co/i infections.
Travelers’ diarrhea in Mexico appears to be a syndrome
caused by a variety of pathogens, the most common of which are
enterotoxigenic E. colt.
ea
Metcalf, T.G. Prospects for Virus Infections in Man and Animals
from Domestic Waste Land Disposal Practices. In “Virus Aspects
of Applying Municipal Waste to Lands” Symposium Proceedings,
edited by LB. Baldwin, J.M. Davidson, and J.F. Gerber. Center for
Environmental Programs, University of Florida, Gainesville (1976),
97-117.
Spray irrigation with sewage effluents is the waste disposal
procedure most likely to cause infections in man and animals. In-
jection of wastes into soils appears to offer the least opportunity
for dissemination of viruses among man and animals.
The likelihood of disease outbreaks in man in the vicinities of
land disposal sites (when disposal is other than by spray irri-
gation) seems remote.
y
Morin, R.A., Keller, J.W., Schaffernoth, T.J. Ozone Pilot Plant
Studies at Laconia, New Hampshire. In “Proceedings of Sym-
posium—New Trends in Water and Sewage Treatment Using Pure
27
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Oxygen and Ozone 1 ” Denver, Colorado, October 1974. U.S. Bu-
reau Reclamation, Engineering Research Center, Denver, Col-
orado (1975), 99-119.
This paper is essentially the same as Mann, R.A., Keller, J.W.,
Schaffernoth, T.J., Paquette, D.R. (1975). Ozone Disinfection Pilot
Plant Studies at Laconia, New Hampshire, J NEW ENG WATER
WORKS ASSN, 89(3):206-4. See 1975 Literature Abstracts.
0
Mosley, J.W. (1975). The Epidemiology of Viral Hepatitis: An Over-
view. AM J MED SCII 270(2):253-70.
Water- and shellfish-transmitted hepatitis A are briefly dis-
cussed within a comprehensive review of infectious hepatitis.
Nestor, I., Costin, L. (1976). Presence of Certain Enteroviruses
(Coxsackie) in Sewage El f uents and in River Waters of Roumania.
J HYG EPIDEM1OL MICROBIOL IMMUNOL, 20(2):137-49.
In studies from 1962 to 1976 in Roumania, coxsackieviruses A
and B were recovered with the gauze pad procedure from 29°/o of
the sewage samples and from 15% of the river water samples test-
ed. Sixty-five samples of aqueduct water yielded two viruses, and
39 samples of sand filter effluents yielded three viruses. In river
water, the frequency of virus recoveries decreased as distance
from the pollution sources increased.
Viruses were recovered by the gauze pad tecnnic. The viruses
were concentrated from gauze pads by the Amberl te method, the
yeast cell method, or the aluminum hydroxide method.
With river and aqueduct waters and sand filter effluents, the
yeast cell concentration method yielded the most viruses.
Niemi, M. (1976). Survival of Escherichia coli Phage T7 in Differ-
ent Water Types. WATER RES, 1O(9):751-5.
Coliphage T7 survived longer in the dark in ground water than
in river water, pulp wastewater, lake water, or brackish water.
At 20 C, 99.9% of the virus was inactivated in 8 to 18 days n
river water, in 20 days in pulp wastewater, in 10 to 16 days in lake
water, and in 10 to 14 days in brackish water.
At 3 C, 90% of the virus was inactivated in 10 to 22 days in riv-
er water, in 20 to 25 days in paper pulp wastewater, in 15 to 30
days in lake water, and in 8 to 30 days in brackish water.
At 3 C and 20 C, less than 5% of the virus was inactivated in
ground water after 32 days of storage.
Differences in the rate of inactivation of the virus occurred,
without apparent pattern, in waters sampled in winter and
summer.
28
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Nikolaeva, TA., Bagdasaryan, G.A., ltskova, A.I., Talaeva, Y.G.
(1975). Hygienic Aspects of Microbial Contamination of Water Ba-
sins. VESTN AKAD MED NAUK, 3:46-52. Russian.
The survival of viruses in waters is discussed briefly within a
review of recreational and other water problems.
Novikov, Y.V. (1975). Hygienic Problems of the Ozonization of
Drinking Water. KAZAN MED ZH, 56(5):77-9. Russian.
The inactivation of enteroviruses by ozone is discussed briefly
in a review of the treatment of drinking water with ozone.
Nupen, EM. Viruses in Renovated Waters. in “Viruses in Water,”
edited by 0. Berg, Hi. Bodily, E.H. Lennette, J.L. Melnick, T.G.
Metcalf. American Public Health Association, Washington, D.C.
(1976), 189-95.
The degree to which tertiary sewage treatment intended to
renovate waters removes viruses is evaluated, and the work yet to
be done in order to meet rigid standards for viruses in renovated
waters is assessed.
Olivares, A.L. (1974). Distribution of Enteroviruses in Spain. Labo-
ratory Methods for Their Study. REV SANID HIG PUBLICA,
48(11):1021-65 Spanish.
This paper was listed in the 1975 edition of these abstracts. A
translation was not then available.
From January 1972 to August 1973, during a period of epi-
demic poliomyelitis, seven strains of polioviruses 1, 2, and 3, one
strain of echovirus 7, one strain of a coxsackievirus B and two
unidentified enteroviruses were recovered from a total of 30 sam-
ples of sewage taken in Madrid province.
Oliver, B.G. Carey, J.H. (1976). Ultraviolet Disinfection: An Alter-
native to Chlorination. J WATER POLLUT CONTRL FED,
48(1 1):261 9-24.
The resistance of Escherichia coli to ultraviolet tight, ozone,
and chlorine is compared with that of several pathogenic enteric
bacteria and viruses within the framework of a review of disin-
fection with ultraviolet light.
29
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8
Otumide, E.A. (1976). The Distribution of Hepatitis B Surface Anti-
gen in Africa and the Tropics: Report of a Population Study in
Nigeria. tNT J EPIDEMIOL, 5(3):279-89.
The incidence of HB 5 Ag by complement fixation tests was
12.6% in an urban high socioeconomic area and in the central
slum area of an African city.
In the slum area, a significant association occurred between
the presence of the antigen and the type of latrine in the house,
the number of blood tests done within two years of the study, and
receipt of injections in a dispensary. In the high socioeconomic
area, a lack of protection from mosquitoes was significantly asso-
ciated with the presence of antigen.
A
Payment, P., Gerba, C.P., Wallis, C., Melnick, J.L. (1976). Methods
for Concentrating Viruses from Large Volumes of Estuarine Water
on Pleated Membranes. WATER RES, 1O(1O):893-6.
Poliovirus 1, seeded into 380 liters (100 gallons) of acidified
seawater (pH 3.5), was concentrated into a final eluate volume of
20 to 100 ml. Fifty percent of the virus was recovered; a concen-
tration of 20,000 to 100,000 was achieved.
The viruses were recovered in a Wallis-Melnick concentrator
equipped with 3- and a 0.45-sm pore size pleated cellulose nitrate
membrane filters. The flow rate through the filters was about 22 li-
ters (6 gallons)/minute. Viruses were eluted from the pleated fil-
ters with glycine buffer (pH 22.5) and reconcentrated by precip-
itation with FeC I 3 or AICI 3 .
The filters still adsorbed viruses after 2,268 liters (600 gallons)
of turbid estuarine water had been passed through them.
V
Peleg, M. (1976). The Chemistry of Ozone in the Treatment of Wa-
ter. WATER RES, 1O(5):361-5.
The dissociation products of ozone in water may be more
powerful oxidizing agents than ozone itself.
The hydroxyl radical (01-1) may be responsible for the high
oxidative and germicidal potentials that occur when ozone is
added to water, but the OH radical preferentially reacts with
organic substances when these are present.
The disinfection capability of other products of ozone dis-
sociation in water (HO 2 , O , O , and 0) are not yet known.
y
Pletri, C., Brelttmayer, J.-P. (1976). Study of the Survival of an En-
terovirus in Ocean Water. REV INT OCEANOGR MED 41-42:77-
84. French.
In laboratory studies at 18 C, poliovirus 1 survived longer in
autoclaved sea water than in filtered sea water, and longer in f ii-
30
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tered sea water than in natural sea water. The sea water samples
were exposed continuously to alternate 12-hour periods of dark-
ness and artificial light.
In 60 days, about 5% of the virus was destroyed in autoclaved
sea water, and about 99.5% of the virus was destroyed in filtered
sea water. About 99.95% of the virus was destroyed En natural sea
water after about 40 days.
eL1 L S
Prozesky, OW. (1975). Viruses in Water. GENEESKUNDE,
17(8):1 62-5. Afrikaans.
The presence of viruses in the water environment, the de-
tection of these viruses, the effect of water purification processes
on them, and the viral standards for water purity are reviewed and
discussed.
11
Quarles, J.R., Jr. Viruses in Water. Legal and Regulatory Decisions
and Dilemmas. In “Viruses in Water,’ edited by G. Berg, H.L. Bodi-
ly, E.H. Lennette, J.L. Melnick, T.G. Metcalf. American Public
Health Association, Washington, D.C. (1976), 209-17.
Recent federal laws impacting USEPA research programs are
recounted and the USEPA research programs in water are de-
scribed against the background of a discussion of water needs
and water pollution problems in the United States.
77
Rao, N.U. Geographic Considerations in Virus Removal in Asia. In
“Viruses in Water,” edited by G. Berg, H.L. Bodily, E.H. Lennette,
J.L. Melnick, T.G. Metcalf. American Public Health Association,
Washington, D.C. (1976), 236-46.
The concentration of enteric viruses in the aquatic environ-
ment in Asia is much greater than in developed countries. The
proportion of children under age 15, where the highest incidence
of excretors occurs, is much higher in Asia than in developed na-
tions. Sewage treatment receives a low priority in the competition
for scarce funds in underdeveloped countries.
People deserve adequate and safe water whether they live in
underdeveloped or developed countries, whether they are poor or
well-to-do. In poor countries, however, progress in this direction,
although noticeable, is slow.
Rao, V.C. (1976). Monitoring, Prevention and Control of Viral Pol-
lution of Water. J INST PUBLIC HLTH ENGRS (IPHE), India,
(3):51 -9.
In India, untreated domestic wastes that contain large num-
bers of human enteric viruses are commonly discharged into Wa-
31
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ter courses, most of which are used as potable water sources by
downstream communities.
Methods for monitoring waters for viruses and for prevention
and control of the pollution of waters with viruses are described
within a review of the waterborne viral problem in India and in
other parts of the world.
e c
Rao, V.C. (1976). Virus Transmission Through Foods. J FOOD SCI
TECHN, 13(6}:287-93.
Methods for recovering viruses from shellfish are discussed
within a review of the problem of the transmission of viruses by
foods.
Reall, D., Caroll, G., Levre, E. (1975). Salmonellae, E coli and
Their Homologous Phages in Seawater. INN SCLAVO, 17(6):842-
7. Ita lIan.
Although coliform bacteria are the most important index of
the degree of fecal contamination of water, other important indi-
ces also exist. The quantitative determination of salmonellae, now
possible with modern procedures, can indicate the extent of con-
tamination with those pathogens and can provide, therefore, a bet-
ter determination of the extent of danger of a water.
Coliphages and phages of Salmonella typhosa and S. ty-
phimurium may also be useful indicators of the degree of con-
tamination of a water. Phages may still be present in water after
the bacteria in which they originated have disappeared.
&
Reeves, W.C., Peters, C.J. (1976). The Seroepidemiology of Reovi-
rus, Coxsackievirus B, and Hepatitis B Virus Infections Among
Panamanian lndian Children, J INFECT DIS, 133(3):268-73.
The rates of infection with hepatitis B virus varied significantly
among Panamanian indian tribes. The prevalance of antibody to
HB 5 Ag was greater in Choco and mainland Cuna Indians than in
Guaymi and island Cuna indians.
In children ten years old or younger, serological studies indi-
cated that infection with group B coxsackieviruses occurred more
frequently in Ghoco and mainland Cuna indians than in Guaymi
or island Cuna indians. Although almost all children tested had
been exposed to reoviruses, mainland Cuna indian children had
the highest geometric mean titers of antibody, followed by Choco,
Guaymi, and island Cuna indians, in descending order.
This parallel in rates of hepatitis B and enteric virus infections
supports the hypothesis that fecal-oral transmission of hepatitis B
virus is important in certain populations.
The village water supplies appeared to be the major environ-
mental difference that distinguished one tribe from another.
32
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1
Reynolds, B.J. An Example of Sewage Disposal by Spray Irri-
gation. In “Viruses in Water,” edited by G. Berg, H.L. Bodily, E.H.
Lennette, J.L.. Melnick, T.G. Metcalf. American Public Health Asso-
ciation, Washington, D.C. (1976), 218-20.
Effluents from a series of stabilization lagoons fed with animal
manure were applied to crops and woodlands which removed nu-
trients and produced effluents from which viruses could not be re-
covered and which met the tertiary treatment requirements of the
Pennsylvania Department of Environmental Resources.
Richards, W.N., Shaw, B. (1976). Developments in the Micro-
biology and Disinfection of Water Supplies. J INST WATER EN-
GRS SCI, 30(4):191-202.
The problems of viruses and pathogenic bacteria in water
supplies are noted briefly in a review of disinfection methods.
i i
Safterman, R.S., Morris, M.-E. (1976). Assessment of Virus Removal
by a Multi-Stage Activated Sludge Process. WATER RES,
1O(5):41 3-20.
In a 50,000 gpd three-stage activated sludge pilot plant seed-
ed with coliphage f 2, 95% of the virus was removed in each the
denitrification and the high-rate activated sludge modules. The
combined unit processes removed more than 99.97% of the virus
from sewage.
The large variations that occurred in the numbers of viruses in
sewage over a 24-hour period demonstrated the importance of ob-
taining temporally-related samples within the treatment system.
A correlation between the numbers of coliphages and the
numbers of enteroviruses in the primary effluent could not be
demonstrated.
Salo, R.J., Cliver, DO. (1976). Effect of Acid pH, Salts, and Tem-
perature on the Infectivity and Physical Integrity of Enteroviruses.
ARCH VIROL, 52:269-82.
At 2 C and 30 C, enteroviruses were more stable at acid pH
levels than at alkaline p 1- I levels. NaCI and other chlorides in-
creased the rate of inactivation of poliovirus at pH 3. The effect of
NaCI was considerably less at pH levels of 4.5 to 7 0 than at pH
levels below 4.5. The RNA of the virion became non-infective as
rapidly as the intact virion did, except at pH 3 in the presence of
MgCI 2 .
Inactivation resulted in alterations to the physical integrity of
enteroviruses. At pH 5 and 7, the RNA of the poliovirus became
hydrolyzed; at pH 3, 5, 6, and 7, the nucleic acid of the virion be-
33
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came susceptible to ribonuclease. Only at pH 3 were virions sensi-
tive to chymotrypsin.
The hemagglutinins of echovirus 7 were destroyed when yin-
ons were inactivated at pH 3, 4. 5, and 6; but, at pH 6 this alter-
ation preceded the loss of infectivity.
The pH of the suspension was a primary determinant of the
mechanism of virus destruction and possibly of the loss of infec-
tivity at 2 C and 30 C.
1 YP
Sattar, S.A., Ramia, S., Westwood, J.C.N. (1976). Calcium Hydrox-
ide (Lime) and the Elimination of Human Pathogenic Viruses from
Sewage: Studies with Experimentally-Contaminated (Poiiovirus
type 1, Sabin) and Pilot Plant Samples. CAN J PUBLIC HLTH,
67(5-6):221 -6.
When added in a quantity sufficient to raise the pH of sewage
to 11.5, lime removed or destroyed most of the 30 to 40 million
PFU of pohovirus 1 seeded into it.
In eight experiments with different samples of sewage, none
of the seeded poliovirus 1 was detected in the supernates. The polio-
virus recovered from the sludges represented only 0.01% of the
input virus. The virus was not recovered from sludges after the
sludges had been stored for 24 hours at 28 C.
The lime process removed viruses equally well at 4 C.
)tp
Sattar, S.A., Westwood, J.C.N. (1975). A Method for the Isolation
of Indigenous Human Viruses from Sludge. IRCS MED SCI, 3:431.
More viruses were eluted from centrifuged sludge solids with
1O% fetal calf serum than with 3% beef extract. Elution of viruses
from sludge solids with 3% lactalbumin hydrolysate yielded rela-
tively few viruses.
AP
Sattar, S.A., Westwood, J.C.N. (1976). Comparison of Four Eluents
in the Recovery of Indigenous Viruses from Raw Sludge. CAN J
MICROBIOL, 22(1O):1 586-89.
In tests with 20 different samples of raw sludge, 3% beef ex-
tract eluted viruses from 15 (75%), and 10% fetal calf serum eluted
viruses from 19 (95%) of the samples. Casein hydrolysate and sac-
talbumi n hydrolysate were ineffective elutants.
The viruses were recovered in BSC-1 cells.
A
Sattar, S..A., Westwood, J.C.N. (1976). Comparison of TaIc-Celite
and Polyelectrolyte 60 in Virus Recovery from Sewage: Devel-
opment of Technique and Experiments with Poliovirus (Type 1,
Sabin)-Contaminated Multilitre Samples. CAN J MICROBIOL,
22(11):1 620-7.
34
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Poliovirus 1 seeded into 100 ml of sewage was recovered by
adjusting the pH of the sewage to 6 and filtering it through either
a layer of PE 60 (400 mg) or through a layer of talc (300 mg)-
Celite (100 mg). The adsorbed virus was eluted from the adsorbants
with 100 ml of 10% fetal calf serum in saline, pH 7.2. The seed
concentration of viruses was 45 to 60 PFU. From 73 to 80% of the
virus was recovered on the PE 60 layers and from 65 to 70% of the
virus was recovered on the talc-Celite layers.
From 33 to 63% of the poliovirus was recovered from talc (15
gm)-Celite (5 gm) layers through which five liter-quantities of sew-
age seeded with 50 to 1.26 x 10 PFU of the virus had been
passed. The virus was eluted with 50 ml of the 10% fetal calf se-
rum in saline.
Except for pH adjustment and prefiltration through two layers
of gauze to remove large solids, no sample pretreatment was
necessary.
With these filtration technics, indigenous viruses have been
recovered from field samples of raw sewage and effluents.
Schaub, S.A.., Sorber, CA. Viruses on Solids in Water. In “Viruses
in Water,” edited by G. Berg, H.L. Bodily, E.H. Lennette, J.L. Mel-
nick, T.G. Metcalf. American Public Health Association, Wash-
ington, D.C. (1976), 128-38.
EMC viruses (rodent enteroviruses), polioviruses, and coli-
phages, adsorbed to solids, retain almost complete infectivity for
cell cultures. Enteroviruses adsorbed to clay are as infective in
vivo as enteroviruses in suspending medium without clay.
Viruses adsorb to colloids and solids in water to a degree dic-
tated by physical-chemical conditions. Adsorption and elution of
viruses to and from solids are affected by the nature of the ad-
sorbing material, the concentration and valence of metal cations
present, the nature of the virus (isoelectric point, whether un-
enveloped or enveloped), pH, the presence of organic and inor-
ganic interfering substances, and time.
Viruses adsorbed to solids assume the dispersal character-
istics of the solids and probably are carried in the water until the
water velocity is low enough to allow the solids to settle. In-
creased water velocity may resuspend the solids and the adsorbed
viruses. Rain reduces the concentration of cations in water and
may thereby release viruses from solids. Organic materials from
domestic and industrial waste effluents and from land runoff may
elute viruses by competitive replacement at adsorption sites.
Generally, all waters reach the oceans. But, the oceans should
not be regarded a final sink for viruses, because infection of man
may occur from ingestion of virus-contaminated shellfish.
Many methods used to concentrate viruses in natural waters
and wastewaters do not recover viruses adsorbed to suspended
matter.
35
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‘ 7
Schmidt, N.J., Ho, HH., Lennette, E.H. (1975). Propagation and
Isolation of Group A Coxsackieviruses in RD Cells. J CLIN Ml-
CROBIOL, 2(3):183-5.
A number of group A coxsackieviruses, including A5 and A6,
multiplied in a cell line designated RD that had been derived from
a human rhabdomyosarcoma. Coxsackieviruses A5 and A6, here-
tofore 1 had been propagated only in suckling mice.
Several group A coxsackieviruses that replicate in other cell
lines multiplied to higher titers in RD cells.
In limited tests, RD cells were slightly less sensitive than suck-
ling mice for recovering group A coxsackieviruses from clinical
specimens.
Group B coxsackieviruses replicated poorly or not at all in RD
cells.
Schwartzbrod, J., Dlxneuf, P., Schwartzbrod, L.., Brochet, J.-C.
Foliquet, J.-M. (1975). Study of the Behavior of Po!iomyelitis Virus
Type 1 in Different Hydrous Media. REV EPIDEM MED, 23(4-
5) :235-52. French.
At 18 C to 24 C, poliovirus 1 survived about 296 days at p 1-I
6.9 in buffer and in buffer that contained soil extract.
Extrapolations from data that extended only to 550 days sug-
gested the virus would survive for 10 to 15 years at 4 C.
The poliovirus concentration in the buffers was more than 10
MPNCU/ml.
y
Sharp, 0G., Floyd, R., Johnson, J.D. (1976). Initial Fast Reaction
of Bromine on Reovirus in Turbulent Flowing Water. APPL ENVI-
RON MICROBIOL, 31(2):173-81.
The destruction of single particles of reovirus by bromine fol-
lowed first-order kinetics. At 2 C, 3-PM of bromine/liter reduced
the plaque titer of the virus at the rate of 3 log 10 units/second at
pH 7. The rate of destruction by bromine of virions in aggregates
of 2 to 10 per clump decreased with time. At a survival level of
10 for single virions, the aggregated preparations lost only 99%
of their plaque titer. In these aggregated preparations, a survival
level of 1O- was reached only after four seconds of exposure to
the bromine.
The destruction rate did not appear to be a simple function of
the size and frequency of viral aggregates even when the aggre-
gates did not contain foreign material.
In unpurified virus preparations (crude freeze-thaw lysates of
infected cells), from 50% to more than 90% of the infectivity was
contained in large, fast-sedimenting aggregates. In polluted wa-
ters, such aggregates probably influenced the rate of destruction
of viruses by disinfectants.
36
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The kinetic disinfection studies were accomplished in a newly
designed device that allowed sampling at one second intervals.
Zr
Shortridge, K S., Hu, L.Y. (1975). Absence of Detectable Hepatitis
B Antigen (HBAg) in the Commercial Oyster, Crassostrea gigas
Thunberg, in Hong Kong. SOUTHEAST ASIAN J TROP MED
PUBLIC HLTH, 6(3):305-7.
Efforts to detect hepatitis B antigen by counter-immu-
noelectrophoresis in commercial oysters (Crassostrea gigas
Thunberg) (or in extracts of oyster tissues including the digestive
diverticula and mantle fluids) grown in polluted Hong Kong waters
were unsuccessful. Tests for the antigen were done monthly over
a one-year period and once hourly during one day in mid-
summer.
Pollution of the oyster beds may have been predominantly of
animal rather than of human origin. A lack of sufficient human
pollution could have accounted for the apparent absence of hepa-
titis B antigen in the shellfish.
U
Shuval, HI. Water Needs and Usage. The Increasing Burden of
Entero viruses on Water Quality. In “Viruses in Water,” edited by 0.
Berg, Hi. Bodily, E.H. Lennette, J.L. Melnick, T.G. Metcalf. Ameri-
can Public Health Association, Washington, D.C. (1976), 12-25.
The history of the problem of viruses in water is recounted
along with the history of research into the problem and its
alleviation.
The author advocates current standards of 0 viruses/ lOU gal-
lons of drinking water, 0 viruses/lU gallons of recreational water,
and 0 viruses/lOU gallons of renovated waters.
The author suggests that these standards should be applied
initially only in cities with populations in excess of 100,000 that
draw their waters from polluted surface sources.
mA
Sigel, M.M., Rippe, D Y., Beasley, A.R., Dorsey, M Jr. Systems for
Detecting Viruses and Viral Activity. In “Viruses in Water,” edited
by 0. Berg, H.L. Bodily, E.H. Lennette, J.L. Melnick, 1.0. Metcalf.
American Public Health Association, Washington., D.C. (1976),
139-5 1.
The methods used for detecting viruses are reviewed.
Sentinal systems for detecting viruses in seawater are de-
scribed. In experimental systems, viruses were retained in the gills
of spiny lobsters, in the gills, intestines, and livers of ap ysia, and
in the liver and intestines of the conch.
37
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Sigel, MM., Rippe, D.F.., King, S.D., Rose, E., Breakenridge, F.,
Dorsey, M., Jr., Watkins, S.L, Jr.,, Dunkley, L., Beasley, A.R.
Survival of Enteric Viruses Under Different Environmental Condi-
tions. In “Virus Aspects of Applying Municipal Waste to Land,”
Symposium Proceedings, edited by LB. Baldwin, J.M. Davidson,
and J.F. Gerber. Center for Environmental Programs, University of
Florida, Gainesville (1976), 53-8.
Polioviruses in the stools of infants and young children who
had been vaccinated orally with live trivalent vaccine ranged in
concentration from 102 to 104.2 lD 50 /gm 10 days after vaccination.
The viruses were stable in extracts prepared in Hanks’ balanced
salt solution for at least 19 days at -20 C, but 90% of the viruses
were inactivated in 12 days at 4 C and 24 C, and 99% were inac-
tivated in five days at 37 C. At 24 and 37 C, the viruses were even
less stable in solid stools.
Coxsackievirus B4 was stable in fecal extracts for at least 10
days at 4 and 24 C, but about 95% of the virus was inactivated in
live days at 37 C.
Echovirus 11 was stable in fecal extracts for at least 28 days,
but the virus was increasingly less stable with rising temperatures.
Contact with paper diapers and urine increased the rate of in-
activation of the polioviruses, but moisture and free air exchange
did not significantly impact survival of these viruses.
y
Slnskey, A.J.., Shah, D., Wright, K.A., Merrill, E.W., Sommer, S.,
Trump, J.G. Biological Effects of High-Energy Electron Irradiation
of Municipal Sludge. In “Radiation for a Clean Environment.” Pro-
ceedings of the International Symposium on the Use of High-Level
Radiation in Wastewater Treatment—Status and Prospects, Mu-
nich, March 1975. International Atomic Energy Agency, Vienna,
Austria (1975), 139-49.
At atmospheric pressure, 99% of Salmonella typhimurium
(R6008), bacteriophage P22, S. typhimurium LT 2, and Escherichia
coli K12, in 0.0067 M phosphate buffer, were inactivated by ap-
proximately 180,000, 75,000, 40,000, and 30,000 rads of electron ir-
radiation, respectively.
Bacteria were destroyed by electron irradiation more rapidly
in blended sludge than in whole raw sludge suggesting that large
clumps protected bacteria from irradiation. Irradiation of bacteria
at 10 C and 35 C produced about the same rates of inactivation in
sludge, but the inactivation of electron-irradiated bacteria in
sludge occurred more rapidly in an atmosphere of oxygen than it
did in an atmosphere of air and more rapidly in air than in
nitrogen.
Irradiations were performed with a compressed gas-insulated
4 MeV Van de Graaff electron accelerator.
38
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Iv k, ., ôervenka, J., Tarab ak, M., Roda, J., Mayerov , A.,
StIlicha, 0., MUo ovI ov , A., Sventekov , S. (1976). The Results
of Examination of Waste Waters Within the Framework of Surveil-
lance of Poliomyelitis in Slovakia in the Years 1968-1972. CS EPI-
DEMIOL, 25(4): 196-204. UkraInian.
From 1968 to 1972 in Slovakia, 1,979 enteroviruses were re-
covered on gauze pads from 4,393 samples of wastewater. Of these
enteroviruses, 647 (32.7%) were polioviruses.
Polioviruses were recovered from 73 of 422 (l7.3%) samples
of wastewater five months after vaccination programs were com-
pleted, but only sporadically in subsequent months.
The genetic markers of polioviruses recovered more than six
months after the beginning of vaccination programs should be
determined.
VP
SivInaki, H.D. Treatment of Sewage Sludge with Combinations of
Heat and Ionizing Radiation (Thermoradiation). In “Radiation for a
Clean Environment.” Proceedings of the International Symposium
on the Use of High-Level Radiation in Wastewater Treatment—
Status and Prospects, MunIch, March 1975. InternatIonal Atomic
Energy Agency, Vienna, Austria (1975), 151-67.
Ninety-nine percent of poliovirus 1 in minimal essential me-
dium was inactivated in about 85 minutes without irradiation at 46
C. in about 200 minutes by 200 krad of gamma irradiation at 23 C,
and in about 40 minutes by 200 krad of gamma irradiation at 46 C.
The anticipated additive effect for the irradiation and the high
temperature would have resulted in inactivation of 99% of the virus
in 60 minutes. Synergism was thus suggested.
Similar synergistic inactivation in sludge occurred with
Bacillus subtilis var. niger spores, coliphage T4, Escherichia colt
and a fecal streptococcus.
137 Cesium was the gamma source.
Slote, L. (1976). Viral Aerosols—A Potential Occupationally Re-
lated Health Threat in Aerated Wastewater Treatment Systems. J
ENVIRON HLTH, 38(5):310-14.
The possible transmission of viruses by aerosols from sewage
treatment plants is discussed.
Smedberg, C.T., Cannon, R.E. (1976). Cyanophage Analysis as a
Biological Pollution Indicator—Bacterial and Viral. J WATER POL-
LUT CONTRL FED, 48(1O):2416-26.
At 25 C, 99.9% of cyanophages (Lyngbya, Phormidium, and
Plectonema-1) in distilled water were inactivated in 20 minutes by
1 mg of chlorine/liter at pH 7.0. Under similar conditions, 99.7% of
39
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Escherichia co/i was inactivated by 0.3 mg of chlorine/liter in 20
minutes.
Field studies have shown that whenever Lyngbya, Phormi-
dium, and Plectonema cyanophages were eliminated, the coli-
forms were too.
The cyanophages are practical and inexpensive viral indi-
cators and may serve as replacements for the coliform test.
77
Sobsey, M.D. Field Monitoring Techniques and Data Analysis. In
“Virus Aspects of Applying Municipal Waste to Land,” Symposium
Proceedings, edited by LB. Baldwin, J.M. Davidson, and J.F.
Gerber. Center for Environmental Programs, University of Florida,
Gainesville (1976), 87-96.
The design of a field monitoring program for enteric viruses in
wastes disposed to the land should consider a variety of site-spe-
cific factors such as type of wastes, nature and extent of treatment
of the wastes, method for applying the wastes to the land, season-
al, climatic, and meteorological conditions, geology, hydrology,
and soil characteristics, vegetative cover, intended subsequent use
of the land, and the topology and population of the area.
Sobsey, M.D. Methods for Detecting Enteric Viruses in Water arid
Wastewater. In “Viruses In Water,” edited by G. Berg, H.L. Bodily,
E.H. Lennette, J.L. Melnick, T.G. Metcalf. American Public Health
Association, Washington, D.C. (1976), 89-127.
With the exception of some raw sewages, enteric viruses in
waters must be concentrated before they can be detected and
quantified by infectivity assay procedures.
Polymer two-phase separation, polyvalent cation salt precip-
itation and some of the simple filter adsorption-elution systems
are methods particularly well suited for small quantities of viruses.
Flow-through filter adsorption-elution systems such as the
Wallis and Melnick portable virus concentrator and ultrafiltration
through anisotropic, polymeric membranes in tangential flow sys-
tems show promise for concentrating small quantities of viruses
from large volumes of highly treated or finished waters that are
low in turbidity and dissolved organic matter.
Good methods for detecting small quantities of viruses in
large volumes of turbid natural waters such as river, lake, and
coastal waters are not yet available.
The use of a nqmber of different cell culture systems permits
the detection of most enteric viruses in water and wastewater. Ad-
equate methods for recovering hepatitis A and B viruses in water
and wastewater are still not available.
40
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Ya
Sorber, C.A. Viruses in Aeroso!ized Wastewater. In “Virus Aspects
of Applying Municipal Waste to Land,” Symposium Proceedings,
edited by L.B. Baldwin, J.M. Davidson and J.F. Gerber. Center for
Environmental Programs, University of Florida, Gainesville (1976),
83-6.
The initial loss of viruses aerosolized during spray irrigation
with wastewater may be as much as 30 to 4 O%. A die-off of 90% of
the remaining ‘,,iruses may be expected every 40 seconds
thereafter.
Under the least desirable meteorological conditions, 99.9% of
the viruses that are aerosolized should be destroyed within 200
meters of the spray nozzle.
Assuming adequate disinfection of the effluents that are
sprayed, large buffer zones may be needed for esthetic purposes
only in the vicinities of fields that are irrigated with sprayed
effluents.
,iiyP
Sproul, O.J. Removal of Viruses by Treatment Processed. In
“Viruses in Water,” edited by G. Berg, H.L. Bodily E.H. Lennette,
J.L. Melnick, T.G. Metcalf. American Public Health Association,
Washington, D.C. (1976), 167-79.
Primary sedimentation sludges contain many viruses and must
be disposed of carefully to prevent contamination of people and
food crops.
In laboratory studies, coagulation of viruses in water with met-
al coagulants has removed 90 to 99% of the viruses. Cationic poly-
electrolytes added with a metal coagulant do not affect the num-
bers of viruses coagulated, but when used alone effectively
remove viruses. Lime can be an especially effective coagulant be-
cause the high pH it produces can be sufficient to destroy viruses.
Viruses are removed from water by sand filtration; the viruses
adsorb to the sand or to particutates trapped in the sand. Filtration
of the flocced waters may remove more than 90% of the previously
unadsorbed virus.
Trickling filters remove relatively small amounts of viruses
from sewage. Activated sludge removes more than 90% when the
process is operating properly.
HOCI is an effective virucide. Chioramines are relatively slow
virucides. Ozone is an effective virucide, but it has a half life of
only 25 minutes.
Anticipated removals of viruses by treatment processes, as-
sumed from laboratory data, suggest that drinking water supplies
developed from contaminated waters may contain viruses.
41
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Sproul, O.J. Standards for Viruses in Effluents, Sludges, and
Ground and Surface Waters. In “Virus Aspects of Applying Munici-
pal Waste to Land,” Symposium Proceedings, edited by L.B. Bald-
win, J.M. Davidson, and JF. Gerber. Center for Environmental
Programs, University of Florida, Gainesville (1976), 131-5.
Data are not available that causally relate viruses in water to
diseases in man, nor are systems available that are sufficiently
sensitive to detect viruses in the concentrations that might be ex-
pected in a good quality effluent from a water renovation plant.
Nonetheless, goals may be established for numbers of viruses
permitted in various waters. To this end 100 PFU/liter may be ac-
ceptable in sewage effluents discharged to adequately diluting
surface waters or to land where the groundwater is treated before
it is consumed. Four-40 PFU/10 gallons may be acceptable for
surface waters.
Sufficient data are not available for establishing goals for
sludges. However, treatment systems are available that can
achieve any degree of inactivation of viruses desired in sludges
and in wastewaters also.
2 ’
Srlramulu, N., Chaudhuri, M. (1976). Dual-Media Coal-Sand Filter
and Its Virus Removal Potential. INDIAN J ENVIRON HLTH,
18(2) :77-86.
Filtration of effluent seeded with coliphage MS2 through dual
media (Giridib bituminous coal and sand) at 9.8 meters/hour (4
gpm/ft 2 ) removed 92% of the virus and 85% of the turbidity from
the effluent.
Symons, G.E., Henderson, K.W. (1976). Disinfection—Where are
We Going? J NEW ENG WATER WORKS ASSN, 90(3):199-208.
The problems of viruses in water are discussed briefly in a re-
view of disinfection of water supplies.
zi
Veitser, V.1., Ryabchenko, V.A., Arbuzova, N.A. (1976). Utilization
of Cation/c Polyelectrolytes for Removal of Viruses from Water.
GIG SANIT, O(3):10-13. Russian.
Two cationic polyelectrolytes, polyvinyl-N-benzyl-tri-methyl-
ammonium chloride (VA-2) and polyethyleneimine (PEI-K), effec-
tively coagulated and removed viruses from tap and river water.
Poiydimethylaminomethyl-N-acryloamide (KF-4), also a cationic
polyelectrolyte, was less effective.
In a concentration of 0.5 mg/liter, VA-2 removed more than
99.9% of poliovirus 1 from tap water, and in a concentration of 1
mg/liter, the polyelectrolyte removed more than 99.999% of coIl-
phage Ti from tap water. Three mg/liter of VA-2 removed less
42
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than 99.9% of coliphages Ti and T2 from river water, but 6 mg/li-
ter of VA-2 removed more than 99.999% of coliphage Ti from the
same water.
In a concentration of 0.5 mg/liter, PEI-K removed more than
99.9% of poliovirus 1 and more than 99.999% of coliphage Ti from
tap water. Three mg/liter of PEI-K removed more than 99.99% of
coliphage Ti and more than 99.5% of coliphage T2 from river
water.
One mg/liter of KF-4 removed only a little more than 90% of
coliphage TI from tap water.
In tap water made turbid by the addition of 50 mg/liter of illite,
4 mg/liter of VA-2 or PE1-K removed more than 99.999% of coli-
phage Ti.
Verstraete, W., Voets, J.P. (1975). Micro-Organisms as Indicators
of Environmental Hygiene: Ecology? Taxonomy and Enumeration.
NATUURWET TYDSCHR (GHENT), 57(2):69-84. Dutch.
The paper was listed in the 1975 edition of these abstracts. A
translation was not then available.
in a short review, several methods for concentrating viruses
from water are briefly described.
/s
von Suzani, C., Hazeghi, p. (i976). Infections Transmitted by Fre-
quenting Swimming Pools. SOZ PRAEVENTIVMED, 21(4):i20-l.
French.
Public swimming pools can be the source of infections with
microorganisms such as Mycobacterium balnei, adenoviruses, en-
teroviruses, the agents of plantar warts, the virus of Molluscum
contagiosum, the chlamydiae of swimming pool-conjunctivitis, and
pathogenic fungi.
Pool water of satisfactory quality by standard tests, may con-
tain pathogenic staphylococci and Pseudomonas aeruginosa.
Effective measures for preventing swimming pool transmission
of diseases include continuous recording of the redox-potential of
the water, limiting the number of visitors to the pool to pool de-
sign specifications, better disinfection of sanitary installations,
regular maintenance of equipment, frequent backwashing of fil-
ters, and exclusion from the pool area of visitors with commu-
nicable diseases.
3,
Wallis, C., Gerba, C.P., Melnick, J.L. Photodynamic Inactivation of
Viruses and Bacteria in Sewage Effluents. In “Viruses in Water,”
edited by G. Berg, H.L. Bodily, E.H. Lennette, J.L. Melnick, T.G.
Metcalf. American Public Health Association, Washington, D.C.
(i976), i80-8.
Sewage effluents may be sterilized by a photoreactive hetero-
tricyclic dye and monochromatic light. Coliforms and poliovirus I
43
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may be sensitized by methylene blue and then destroyed by ex-
posure to a source of visible light that emits energy at the wave-
length of light which the dye absorbs. The logistics of photo-
dynamic inactivation appear to be practical for wastewater
disinfection.
Wallis, C., Stagg, C.H., Farrah, S.R., Melnick, J.L. Problems Re-
lated to the Concentration of Viruses from Large Volumes of
Wastewater. In “Virus Aspects of Applying Municipal Wastes to
Land.” Symposium Proceedings, edited by LB. Baldwin, J.M.
Davidson, J.F. Gerber. Center for Environmental Programs, Uni-
versity of Florida, Gainesville (1976), 37-44.
The optimal method developed in our laboratory for concen-
trating viruses from large volumes of sewage effluents involves
four steps. These are: (1) removal of large particles from the efflu-
ents with clarifying filters; (2) adsorption of viruses in the clarified
effluents to membrane filters at low pH; (3) elution of adsorbed
viruses from the membranes with a buffer solution at high PH; and
(4) reconcentration of the viruses into a small volume for assay.
Certain problems are associated with each of these steps. (a)
When inorganic or organic gels are present in an effluent, viruses
may be adsorbed on or trapped within them and be removed by
the clarifying filters. (b) The membrane filters used for adsorption
of viruses may clog. (C) Elution of adsorbed viruses requires high
pH, but the elution time must be short to prevent inactivation of
viruses. With clogged filters, the elution time is extended and
some viruses may be inactivated. (d) The membrane filters adsorb
organic compounds from water samples which may be eluted with
the viruses. These organic compounds form flocs at low pH that
clog the small filters used for reconcentration of viruses.
These problems can be resolved. Samples can be clarified at
high pH with tetrasodium ethylenediaminoacetic acid so that
viruses are not removed by clarifying filters. A melamine-epoxy-
fiberglass membrane filter that resists clogging by water samples
at low pH may be used. Eluates that cannot be reconcentrated on
small diameter membranes because of organics eluted with the
viruses from the clarifying filters and virus-adsorbing filters may
be reconcentrated by an aluminum hydroxide gel technic.
)tp
Ward, R.L., Ashley, CS. (1976). Inactivation of Poliovirus in Di-
gested Sludge. APPL ENVIRON MICROBIOL, 31(6):921-30.
Poliovirus, seeded into anaerobically digested sludge incu-
bated at temperatures of 28 C and 4 C, was fully recovered from
the sludge, but infectivity of the virus decreased in proportion to
the time and temperature of incubation. The rate of inactivation of
the virus ranged from greater than 90%/day at 28 C to about 90%/
5 days at 4 C.
44
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The RNA of inactivated virions appeared to have been dam-
aged and had an average sedimentation value about 70% of that of
RNA from infective polioviruses. Since the specific infectivity of
RNA extracted from polioviruses recovered from sludge was di-
rectly proportional to that of the particles from which the ANA had
been extracted, loss of infectivity was probably due to inactivation
of RNA.
Some breakdown occurred also in the two largest proteins in
the inactivated virions. Thus, inactivation of polioviruses in di-
gested sludge may result from cleavage of viral proteins and sub-
sequent damage to virion RNA.
Virucidal activity was not present in raw sludge; thus, the
component of digested sludge responsible for inactivation of the
poliovirus appears to be a product of the digestion process.
yp
Ward, R.L., Ashley, C.S., Moseley, R.H. (1976). Heat Inactivation of
Polio virus in Wastewater Sludge. APPL ENVIRON MICROBIOL,
32(3) :339-46.
Raw sludge protected the plaque-forming ability of polio-
viruses 1 and 2. In low concentrations at relatively low tempera-
tures, digested sludge protected polioviruses nearly as well as
raw sludge did. At higher temperatures, the rate of inactivation of
the polioviruses accelerated in digested sludge.
The difference in the protective capability of raw and digested
sludges was not accounted for by loss of protective material in di-
gested sludges. The protective component was present in the sol-
ids of digested sludges and in those of raw sludges. A virucidal
agent was produced during digestion of sludges which when
added to the solids of either raw or digested sludges reversed the
protective potential of those solids during heat treatment of
poliovi ruses
50
Waterson, A.P. (1976). Infectious Particles in Hepatitis. ANN REV
MED 27:23-25.
The etiological agents of hepatitis A and hepatitis B are re-
viewed in some detail.
A rP
Wellings, F.M., Lewis, A.L., Mountain, C.W. (1976). Demonstration
of Solids-Associated Virus in Wastewater and Sludge. APPL ENVI-
RON MICROBIOL, 31(3):354-8.
Many of the viruses in wastewaters, sludges, and soils, are
solids-associated. In samples of sewage influent, effluent, and
chlorinated effluent, from 16 to 100% of the total quantities of
viruses recovered were solids-associated. By concentration with
polyethylene glycol, viruses were recovered from the 3onicated
solids of fully digested sludges (60 days at 34 C), from sorncated
45
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sand taken from the site of a sewer lead, and from sonicated dried
sludge cake and mud collected 900 meters downstream from a
sewage disposal site.
These data underscore the importance of processing solids as
a part of any technic for recovering viruses from environmental
waters.
To liberate viruses, sludges in elutant (3°Io beef extract) were
sonicated (100 W for 15 minutes in a rosette cooling cell, Sonifier
Model W 350, Ultrasonic, Inc.), stirred mechanically at 4 C for 18
hours, or extracted with fluorocarbon. The three methods pro-
duced equal recoveries.
Wellings, F.M., Lewis, A.L., Mountain, C.W. Viral Concentration
Techniques for Field Sample Analysis. in “Virus Aspects of Ap-
plying Municipal Waste to Land,” Symposium Proceedings, edited
by L.B. Baldwin, J.M. Davidson, and J,F. Gerber. Center for Envi-
ronmental Programs, University of Florida, Gainesville (1976), 45-
51.
No one virus concentration technic can be used effectively
with all field samples. The membrane adsorption system appears
to be the most promising technic for large samples of water, but
viruses must be eluted not only from the virus adsorbing filters but
from the clarifying filters used in the system also.
Al must not be used in these systems when the waters that
are processed contain large quantities of phosphates, because
viruses bind irreversably to AIPO 4 .
Westwood, J.C.N., Sattar, S.A. The Minimal Infective Dose. In
“Viruses in Water,” edited by G. Berg, H.L. Bodily, E.H. Lennette,
J.L. Melnick, T.G. Metcalf. American Public Health Association,
Washington, D.C. (1976), 61-9.
Laboratory studies provide strong evidence that one virus
particle can establish infection in a single cell in culture and in a
mammalian host. Extreme dilution of viruses in the environment
reduces the probability of infection of individuals in an exposed
population, but does not eliminate it. Since many viruses are sta-
ble in the environment for long periods of time, the effect of viral
pollution of water must be a low-grade seeding of the population
with the production of endemic foci of infection in the community.
The real question to be answered is whether such endemic
foci are of sufficient importance to the community to warrant ma-
jor action.
WHO Expert Committee on Fish and Shellfish Hygiene, Commit-
tee Report. Fish and Shellfish Hygiene. WHO Technical Report
Series, FAO AND WHO, Geneva, Switzerland (1974), 3-62.
46
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The transmission of viral diseases to man by shellfish is dis-
cussed briefly within a broad review of fish and shellfish hygiene.
WHO Scientific Group on Virus Diseases. (1 975-6). The New Pro-
gram of the World Health Organization in Medical Virology.
INTER V1ROL, 6(3):1 33-49.
The viruses that may be transmitted with water, especially the
hepatitis viruses, the gastroenteritis viruses, and the enteroviruses,
are discussed within an overall review of viral diseases.
Wiesmann, E. (1975). Prophylaxis of Virus Hepatitis. GASTROENT
FORTBILDUNGSK PRAX, 5:141-8. German.
The waterborne transmission of hepatitis A is noted within a
discussion of the viral hepatitites and their treatment.
Ii
Zelepukha, S.L, Zemlerub, l.L., Zayakin, Z.R. (1975). Removal of
Microorganisms from Water by Contact Coagulation. MIKROBIOL
ZH, 37(5):648-52. Ukrainian.
Staphylococcus aureus, Escherichia co/i, and a coliphage
were removed from water with eaual effectiveness by coagulation
with FeCI 3 , FeSO 4 , and Al 2 (S0 4 )3 doses up to 40 to 50 mg/liter.
Fe 2 (S0 4 )3 was less effective. A1 2 (S0 4 ) 3 , unlike the other reagents,
was easily overdosed, and this resulted in reduced removal of the
microorganisms and the virus from the water.
The effectiveness of removal of the microbes decreased in the
following order: S. aureus, E. co/i, coliphage. The effectivness of
removal of the S. aureus from the water was inversely related to
its initial concentration.
A.p.
Zotova, V.1 Myshliaeva, L.A. (1976). Quantitative Determination of
Enteroviruses in Waters. GIG SANIT, O(3):63-5. Russian.
Seeded poliovirus 1 was concentrated from sewage and from
autoclaved river and tap water by adsorption of the virus onto AV-
17 (an anion exchange resin) at pH 5 and elution of the virus from
the resin with 0.5 M phosphate buffer at pH 8.2. The extent of ad-
sorption of viruses onto the resin decreased with increased pollu-
tion of the water; 12% of the virus was adsorbed from sewage and
3 9% was adsorbed from tap water. The extent of adsorption of the
virus was inversely related to the concentration of seeded virus.
With this anion exchange technic, 15 to 30 viruses/liter were
recovered from river waters, 980 to 2,320 viruses/liter were recov-
ered from mechanically and biologically treated sewages, and 100
to 350 viruses/liter was recovered from chlorinated effluents.
47
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AUTHOR INDEX
Author
Aizen, M.S 1
Akin, E.W 1,18
Albano, A 2
Alexandre, D 2
Amirhor, P 11
Arbuzova, N.A 42
Ashley, C.S 44,45
AVIO. C.M 6
Bachmann, P.A 2
Baydasaryan, G.A 29
Beamer, P 5
Beasley, A.R 37,38
Beltort, G 2
Bell, R.G a
Bember, V.E. 15
Benton. W,H 1
Berry, SA 3
Beytout, U 23
Biedermann, N 20
Bitton, G 4
Blancard, A 2
Blum, A.L 22
Bonc larenko, V.1 4
Bouwer, H 4,15
Breakenridge, F 38
Breittrnayer, J.-P 30
Brochet, J.-C 36
Brown, ER 5
Buium, I 20
Buras, N 5
Ca llaghan, 0
Cannon, R.E 39
Caraway, C.T 25
Carey, J.H 29
Car lson, S 5 ,6
Caroli, G 6 ,32
Oervenka, J 39
Charrel, J 2
Charrier. F 23
Chaudhuri, M 42
Chian, E.S.K 12
CiIlié, G.G 6
Clarke, N.A 18
Cline, G.B 1
Cliver, D.O 6,33
Cochran, K.W 12
Costin, L 28
Couch, J.A 7
Courtney, L 7
Cramer, W.N 8
Craun, G.E a
Creech, W.B 27
Dartevelle, Z
Page De Donato, S 2
Derbyshire, J.B 9
Desmet, L 3
Dienstag, J.L 9
Dixneuf, P 36
Dobrowo lska, H 19
Dolowy, W.C
Dor, I 9
Dorsey, M., Jr 37,38
Dubo se, S.M 10
Dunkley, L 38
DuPont, H.L 10
Earampamoorthy, S 10
Engelbrecht, R,S 11
Epp, C 11,12
Fanniri, K.F 12
Farley, C.A 12
Farooq, S 12
Farrah, S 13
Farrah, S.R 13,44
Fattal, B 14,21
Feeley, J.C 27
Floyd, A 14,36
Foliguet, J.-M 36
Fox, J.P 14
Freytag, B 15
Gangarosa, E.J 27
Gannon, J.J 12
Gassner, M 22
Gerba, C.P 13,15,22,26,30,43
Gevaudan, P 2
Gifford, G.E 4
Gilbert, J 24
Gilbert, R.G 15
Globa, LI 15
Goldtield, M 16
Goyal, S.M 13
Grabow, W.0.K 16
Grebenyuk, V.D 15
Greenberg, S
Grigoryeva, L.V 4
Gude, G 17
Gulian, C 2
Gust, I.D 9
Guy, M.D 17
Gvozdyak, P.1 15
Hásselbarth, U 5 ,6
Hattingh, W.H.J 17
Hazdra, J.J
Hazeghi, P 43
Henderson, K.W 42
Hetrick, F 24
Hill, B.J 17
8 Hill, W.F., Jr 1,18
48
-------
Myshliaeva, L.A . 47
Ho, H.H 36
Hobbs, M.F 26
Hoehn, R.C 18
Hostovesky, T 21
Hu, L.Y 37
Hughes, D.E 18
Hughes, J.M 8
ltskova, A.l 29
Jakubowski, W 18
Jankowski, M 19
Jarzabek, Z 19
Johnson, J.D 14,36
Johnson . P.1 19
Kalitina, T.A 19
Katter, 5 , 5 19
Kantoch, M 19
Kapikian, A l 27
Katzenelson, E 2,14,20,21
Kawata, K 8
Keith, L 5
Keller, J.W 27
Kiker, C.F 21
King, S.D 38
Koff, R.S 10
Krejs, G.S 22
Krusé, C.W 8
Lance, J.C 22
Larkin, E ,P 22
Laveran, H 23
Lemke, H.S 23
Lennette, E.H 23,36
Levre, E 32
Lewis, A.L 45,46
Lewis, M.J 17
Lewis, N.F 24
Lo, S 24
Lucas, C.R 9
Lund, E 24,25
McCabe, Li 8
Mclver, J.D 17
Mackowiak, P.A 25
Mahdy, M.S 25
Malina, J.F., Jr 26
Mallet, M.N 2
Masterson, N 4
Mayerová, A 39
Maynard, J.E 26
Meinick, J.L 13,15,22,26,30,43,44
Merrill, E.W 38
Merson, M.H 27
Metcaif, T.G 27
Milistein, C.H 19
Milogovi ov& A 39
Moore, B.E 10
Morin, R.A 27
Morris, G.K 27
Morris, M.-E
Moseley, R.H 45
Mosley, J.W 28
Mountain, C.W 45,46
Nestor, I 28
Niemi, M 28
Nikolaeva, l .A 29
Noton, 6.G 3
Novikov, V.V 29
Nupen, E.M 17,29
Olivares, A.L 29
Oliver, B.G 29
Olumide, E.A 30
Pancorbo, 0 4
Payment, P 30
Peteg, M 30
Peters, C.J 32
Pietri, C 30
Pille, EAR 1
Portnoy, B.L 25
Prozesky, O.W 31
Purcell, R.H 9
Quarles, JR., Jr 31
Ramia, S 34
Rao, N.U 31
Ftao, V.C 31,32
Reali, D 32
Reeves, VV.C 32
Reynolds, B.J 33
Rice, R.C 15
Richards, W.N 33
Rippe, D.F 37,38
Roda, J 39
Rose, E 38
Rotem, 2
Rotmistrov, M.N 15
Ryabchenko, V.A 42
Sack, D.A 27
Sack, RB 27
Safferman. R.S 33
Sagik, B.P 10
Salo, cmi 33
Sattar, S.A 34,46
Schaffernoth, T.J 27
Schaub, S.A 35
Schechter, H 9
Schmidt, N.J 36
Schwartzbrod, J 36
Schwartzbrod, L 36
Shaffer, P.T.B 13
Shah, 0 38
Sharp, D.G 14,36
Shaw, B 33
Shortridge, K.F 37
Shuval, H.l 9,20,37
Sigel, M.M 37,38
Sinclair, T 5
Sinskey, A.J 23,38
Siväk, 39
Sivinski, H.D 39
49
-------
Slote, L. 39
Srnedberg, C.T. 39
Sobsey, M.D 40
Sohn, F.W 5,6
Sommer, S 38
Sother, C A 35,41
Spendlove, J.C 12
Sproul, O.J 41,42
Sriramulu, N 42
Stafford, D.A 18
Stagg, Cl-I 44
Stilicha, 0 39
Sullivan, R 22
Summers, M.D 7
Sventeková, S 39
Symons, G.E 42
Talaeva, Y.G 29
Tarabëák, M 39
Tierney, J.T 22
Trump, J.G 38
Veitser, V.1 42
Verstraete, W 43
Voets, J.P 43
von Suzani, C 43
Wallis, C 13,15,26,30,43,44
Ward, R.L 44,45
Waterson, A.P 45
Watkins, S.L., Jr 38
Wellings, F.M 45,46
Wells, JO 27
Westwood, J.C.N 34,46
WHO Expert Committee 46
WHO Scientific Group 47
Wiesmann, E 47
Wong, D 9
Wright, K.A 38
Zayakin, Z.R 47
Zelepukha, S.l 47
Zemlerub, I.L 47
Zotova, V.1 47
50
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SUBJECT INDEX
Each subject category listed on this page is coded by a Greek
letter to simplify the identification of abstracts that contain in-
formation that relates to that category. To use the subject index,
note the pages listed for the category, and read those abstracts
marked by the Greek letter that codes for the category.
Abstracts that are marked by more than one Greek letter con-
tain information in more than one subject category.
Code Subject Page
a Aerosols 12 ,27,39,41
/3 Bathing Waters 15,43
y Disinfection 5,6,8,11,12,14,17,18,20,23,24,25,26,
27,29,30,33,34,36,38,39,41,42,
43,44,45
8 Epidemiology 1,5,8,9,10,14,16,17,19,20,
22,23 ,25,26,27,28,30,32,47
(Fish s
C Food 4,19,22,32
General 17,19,31,33,36,40
0 Hepatitis A 1,8,9,10,14,16,22,25,26,28,45,47
Indicators 3,32,37,39
,< Infective Dose 2,46
X Methodology .. 2,13,14,18,19,20,21,23,30,34,37,40,44,45,46,47
a Recovery from the Environment 5,6,8,9,17,28,29,31,39,47
v Removal by Treatment Processes 2,4,6,9,15,17,18,24,
26,29,31,33,34,41,42,47
e Review 1,2,4,6,8,10,14,15,16,18,19,22,23,24,
25,26,27,28,29,31,32,33,35,37,39,40,41,42,
43,44,45,46,47
o Rotaviruses 10
i i Shellfish 1,7,9,12,17,19,22,25,32,37,46
p Sludges 6,11,12,21,23,25,26,34,39,41,44,45
o Soils 4,10,15,22
r Solids 6,35,45
u Solid Wastes 11,23,38
* Standards 31,37,42
Survival 1,2,3,22,24,28,29,30,33,36,38,41
51
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TECHNICAL REPORT DATA
(Meow trod lasmictions on the revene &f completing)
1. REPORT NO. 2-
EPA-6 00/9-77-028
3. RECtPIENTS ACCESSION NO.
4. TITLE AND SUBTITLE
.
Viruses in Waste, Renovated and Other Waters
1976 Literature Abstracts
5. REPORT DATE
August 1977 (Issuing date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Gerald Berg, Editor
F. Dianne White, Editorial Assistant
I. PERFORMING ORGANIZATION REPORT NO.
I. PERFORMING ORGANIZATION NAME AND ADDRESS
Biological Methods Branch
Environmental Monitoring Support Laboratory-tin.
ERC, Cincinnati, USEPA
10. PROGRAM ELEMENT NO.
1BD7 13
11. CONTRACT/GRANT NO.
In House
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Monitoring Support Laboratory-Cm.
Office of Research and Development
U. S. Environmental Protection Agency
Cincinnati, Ohio 45268
13. TYPE OF REPORT AND PERIOD COVERED
Literature Review - 1976
14. SPONSORING AGENCY CODE
EPA’6OO ’ 6
/ ‘ -
15. SUPPLEMENTARY NOTES
It ABSTRACT
The volume comprises the abstracts of the published papers and books on
viruses in waste, renovated, and other waters for 1976.
7. KEY WORDS AND DOCUMENT ANALYSIS
- OESCRIPTORS
b.IDENTIPIERS/OPEN ENDED TERMS
C. COSATI Ficid/Group
*Viruses, Waste water, Water, Sewage
l3B
‘B, DISTRIBUTION STATEMENT
Release Unlimited
19. SECURITY CLASS (This Report)
UNCLASSIFIED —
21. NO. OF PAGES
, ‘
20. SECURITY CLASS (Thts page)
UNCL&SSIFIET)
22. PRICE
EPA PornI 2220.1 (9-13)
*U.S.GOVERRMUITPRlN11NGOfFIct 1917— 757-133/1605
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