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LITERATURE ABSTRACTS
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U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Research and Monitoring
National Environmental Research Center
Cincinnati, Ohio

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VIRUSES IN WASTE,
RENOVATED,
AND OTHER WATERS
Editor. Gerald Berg, Ph.D.
Editorial Assistant. F. D/anne White
1971
Virology Section
Advanced Waste Treatment Research Laboratory
Office of Research and Monitoring
National Environmental Research Center
Environmental Protection Agency
Cincinnati, Ohio

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1971
VIRUSES IN WASTE, RENOVATED, AND OTHER WATERS
Adamczyk, B., Kiehl, W., Adamczyk, G., Wandel, I. (1970). Epidemiology and
Virology of an Aseptic Meningitis Outbreak in Berlin Associated with
Echovirus 30. DTSCH GESUNDEITSW, 25<11>:2144-8. German.
One-hundred cases of aseptic meningitis occurred in Berlin in the sum-
mer of 1969. Children between 3 and 10 years of age comprised 60% of the
cases. Only 7% of the patients were over 30 years of age.
The outbreak began during an unusual heat wave in July and August.
Outdoor swimming pools appeared to be the source of infection.
Echovirus 30 was recovered from patients much more frequently than
any other virus.
Akin, E. W., Benton, W. H„ Hill, W. F., Jr. Enteric Viruses in Ground and
Surface Waters; A Review of Their Occurrence and Survival. In "Virus and
Water Quality: Occurrence and Control," edited by V. Snoeyink. Proceedings
of the Thirteenth Water Quality Conference, February 1971. University of
Illinois Bulletin, 69 (1971), 59-74.
Enteric viruses occur in surface waters throughout the world. These
viruses are shed in the feces of infected men and animals and enter water
systems by way of soil runoff and sewage, both treated and untreated.
Viruses absorb to soil particles and may be removed before they reach
ground water, but both contaminated surface and ground water have been
implicated in infectious hepatitis outbreaks. The incidence of endemic virus
disease transmitted through surface waters is difficult to determine because of
the large percentage of asymptomatic infections which occur with these viruses.
Babov, D. M., Gubenko, L. T., Muromtseva, A. A., Yarotskaya, N. E. (1971).
Enterovirus Circulation in the Environment and Among the Population of
Odessa. GIG SANIT, 36:54-7. Russian.
Echoviruses 3, 6, and 7 and coxsackieviruses B3 and B5 circulated
throughout the year among the population and in the sewage of Odessa from
1966 to 1968, These serotypes were detected in sewage when they were
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recovered from 1.9 to 4.9% of the population. Viruses were recovered from
sewage by the gauze pad method,
Bagdasaryan, G. A. (1970). Viability of Enteroviruses in Waters with Different
Degrees of Pollution. ZH MICROBIOL, 47(8): 96-8. Translation presently not
available. Russian.
Bagdasaryan, G. A., Abieva, R. M. (1971). Survival of Enteroviruses and
Adenoviruses in Water. GIG SANIT, 36:10-14. Russian.
Survival in water of enteroviruses and adenoviruses was influenced by
temperature and the extent of contamination of water by bacteria. Strains of
adenovirus 5 survived for the longest periods. In tap water, these strains
survived for 63 days at 18 to 20 C, and for longer periods at 4 to 6 C.
Baumann, E. R. (1971). Lawsuit Involving Source of Hepatitis Outbreak.
HSMHA HEALTH REP, 86(1): 19-20.
A lawsuit and its outcome is described in which plaintiffs failed to
collect damages for contracting infectious hepatitis, allegedly the result of
negligence of a landlord in providing a contaminated well-water supply. A
septic tank had been installed only 35 feet from the well, 15 feet closer than
permitted by the state plumbing code. The court found that fecal contamination
of the well had not been demonstrated, and that unsanitary conditions of the
premises and personal contact transmission were the most likely causes of the
outbreak.
Berg, G. An Integrated Approach to the Problem of Viruses In Water. In
Proceedings of the National Specialty Conference on Disinfection, July 1970.
ASCE, New York, New York, (1971), 339-64.
Large quantities of viruses of human origin are excreted into sewage, and
thus abound in our rivers and streams. Viruses have been detected at water
intakes and large numbers apparently enter intakes daily. Since ingestion of a
small amount of virus is likely to produce infection rather than disease, the
inability of epidemiologists to detect waterborne transmission may be the
result of preoccupation with clinical illnesses rather than infection in index
cases.
Viruses are removed from sewage effluents and from raw waters by most
biological, chemical, and physical treatment procedures, but never completely.
Treatment procedures therefore constitute adjunctive removal, complete virus
removal being dependent upon terminal disinfection.
No single disinfectant is capable of adequately disinfecting waters of all
quality. The chemical quality of effluents and the toxicity to aquatic life of
disinfectants and the compounds they form in water are important determi-
nants in the choice of a water disinfectant.
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Berg, G. (1971). Integrated Approach To Problem of Viruses In Water. J SAN
ENGIN DIV, ASCE, 97:SA6:867-82.
This paper is a somewhat updated version of the ohe above. It includes a
proposal that the primary standard for microbiological acceptability of
renovated and other potable waters should be manifested in disinfectant
concentrations and that bacterial and viral determinations, which are usually
unavailable until waters are consumed, should serve in a backup status.
Berg, G. Removal of Viruses from Water and Wastewater. In "Virus and Water
Quality: Occurrence and Control," edited by V. Snoeyink. Proceedings of the
Thirteenth Water Quality Conference, February 1971. University of Illinois
Bulletin, 69(1971). 126-36.
There is not yet available a method that completely recovers all viruses
from sewage.
Primary settling, which is used in most communities, removes viruses
inefficiently. Biological treatment methods such as activated sludge, stabilization
ponds, and trickling filters effect some virus removal but with widely varying
results. Chemical-physical treatment may be more uniformly effective in virus
removal.
As costs become a little less important, and the quality of our waters a
little more important, chemical and physical treatment procedures may find
increasing application both in treating raw sewage and secondary effluents
especially when total renovation is the goal.
Berg, G., Dahling, D. R., Berman, D. (1971). Recovery of Small Quantities of
Viruses from Clean Waters on Cellulose Nitrate Membrane Filters. APPL
MICROBIOL. 22(4):608-14.
Small amounts of viruses were recovered quantitatively from large
volumes of buffered, distilled water by adsorbing the viruses onto cellulose
nitrate membrane filters (0.45 nm pore size) and eluting the adsorbed viruses
in 3% beef extract under extended sonic treatment. Complete recovery of
poliovirus 1, echovirus 7, and coxsackievirus B3 resulted when less than 100
PFU were added to t-liter quantities of water. Recoveries of reovirus 1 were
almost as good. Preliminary studies indicated that good recoveries could be
made from 25-gallon quantities of water. The method described was efficient
in waters of high quality and may be useful for recovering viruses in renovated,
and perhaps in tap waters, but not in waters containing certain organic matter
unless that matter is first removed.
Borisov, L. B., Khan-Fimina, V. A., (1970). Biological Characteristics of the
Phages of h'nteropathogenic Escherichia coli. Report 4. Biological Properties of
Phages Lysing /
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described. The phages differed in antigenic properties, temperature tolerance,
and morphology. On the basis of serology and host range, two phages were
classified in Escherichia coli group 026 and two in E. coli group 0124.
Braga, A., Pagano, A. (1970). Hygienic and Sanitary Aspects of Sea-Water
Pollution. IG MOD, 63:227-48. Italian.
Infective organisms in sewage that pollutes seawater are a hazard to
bathers and contaminate shell fish. Infectious hepatitis has resulted from the
consumption of raw clams.
Brunner, D. R., Sproul, 0. J. (1970). Virus Inactivation During Phosphate
Precipitation. J SAN ENG1N DIV, ASCE, 96:SA2:365-79.
Large quantities of poliovirus 1 were removed during precipitation of
phosphate in distilled water and in domestic wastewater effluent by either
calcium or aluminum.
In distilled water and in domestic wastewater effluent containing equal
molar concentrations of Af"* and P04=, precipitation with aluminum at pH 5.0
effected 90% poliovirus 1 removal when 24 mg/liter P04= were precipitated,
and 98% poliovirus 1 removal when 30 mg/liter P04= were precipitated.
In distilled water at pH 11, calcium effected 97% virus removal when 37
mg/liter P04= were precipitated. In domestic wastewater effluent, calcium
treatment effected 94% virus removal when 44 mg/liter P04= were precipitated.
Removals of viruses by these two phosphate reduction processes were
predictable by a Freundlich isotherm. Poliovirus 1 removals were affected by
hydrogen ion concentration in each process. Alum treatment removed more
viruses than lime treatment of a comparable wastewater.
Canzonier, W. J. (1971). Accumulation and Elimination of Coliphage S-l3 by
the Hard Clam, Mercenaria mercenaria. APPL MICROBIOL, 21(6):1024-31.
Hard clams (Mercenaria mercenaria) exposed to low levels of coliphage
S-l 3 (7 PFU/ml) in running seawater for several days accumulated titers in
tissues from two to more than 1,000 times the levels to which they had been
exposed. Accumulations of Escherichia coli were comparable. Clams polluted
with relatively low levels of E. coli eliminated most of the bacteria in 24 to 48
hours. The virus, however, persisted for days to weeks even under conditions
ideal for clam activity when temperatures were below the inactivation threshold
for the virus. Most of the accumulated virus was sequestered in the digestive
gland. These sequestered particles were refractory to those mechanisms
responsible for elimination of bacteria.
The need for caution in evaluating the efficiency of shell fish depuration
processes is thus underscored when only a bacterial criterion is used as a
monitoring system.
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Chambers, C. W. (1971). Chlorination for Control of Bacteria and Viruses in
Treatment Plant Effluents. J WATER POLLUT CONTRL FED, 43(2):228-41.
Destruction of bacteria and viruses in treatment plant effluents are
reviewed.
Chang, P. W., Liu, O. C., Miller, L. T., Li, S. M. (1971). Multiplication of
Human Enteroviruses in Northern Quahogs. PROC SOC EXP BIOL MED,
136(4): 13804.
Proflavine-labeled poliovirus 1, echovirus 1, and coxsackievirus B3 did not
multiply in Northern quahogs. Virus isolated from the quahogs after inoculation
apparently represented persisting input virus.
Chang, S. L. Modern Concept of Disinfection, In Proceedings of the National
Specialty Conference on Disinfection, July 1970. ASCE, New York, New York
(1971), 339-64: Also in J SAN ENGIN DIV, ASCE. 97: SA5-.689-707, 1971.
Concepts of disinfection are founded on the physical chemistry of
disinfectants, the cytochemical natures and physical states of microorganisms,
the reactions of disinfectants and microorganisms as rate processes, and the
quantitative effects of factors such as pH and temperature.
Disinfectants may be grouped as oxidizing agents, heavy metal cations,
quaternary ammonium and pyridinium compounds, gaseous agents, and
physical agents. The mode of action of each group of agents is different.
The difference in the structure of amoebic cysts, vegetative bacteria,
and viruses accounts for the difference in their resistance to disinfectants.
The presence of organisms in aggregates is probably an important cause
for aberrations in survival curves.
Cherkinsky, S. N., Lovtsevich, E. L., Ryabchenko, V. A. (1971). The
Sanitary Significance of Escherichia coli in Water Decontamination of Entero-
viruses. GIG SANIT, 36-.7-1G. Russian.
The value of Escherichia coli as an indicator for enteroviruses in water
undergoing disinfection depends upon the relative amounts of the bacteria
and viruses present.
Cliver, D. 0. Viruses in Water and Wastewater: Effects of Some Treatment
Methods. In "Virus and Water Quality: Occurrence and Control," edited by V.
Snoeyink. Proceedings of the Thirteenth Water Quality Conference, February
1971. University of Illinois Bulletin, 69 (1971), 149-58.
A polycationic primary coagulant did not remove significant quantities
of enteroviruses from water and wastewater. Viruses were removed by
activated carbon, but a good deal was still present in the effluents from the
carbon columns.
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Oliver, D. O. (1971). Transmission of Viruses Through Foods. CRIT REV
ENVIRON CONTR, 1(1):551-79.
Transmission of viruses by shellfish and contaminated processing waters
are reviewed as part of a general review on foodborne transmission of viruses.
Cookson, J. T. (1969). Mechanism of Virus Adsorption on Activated Carbon.
J AMER WATER WORKS ASSN, 61(1 ):52-6.
Sites on activated carbon that adsorb bacteriophage T4 are probably
carboxyl groups or lactones. Adsorption on carbon can be completely blocked
by esterifying these groups.
Amino groups on the virus apparently adsorb electrostatically to
carboxyl groups on the carbon. The pH range of strong adsorption rules out
participation by ionized forms of sulfhydryl, guanidino, and phenolic groups in
the electrostatic binding. Ionized imidazolyl groups are probably too few to be
significant.
As pH decreases from the optimum of 7, adsorption rates decrease,
probably because viral tail fibers on the tail sheath become unable to attach to
carbon sites.
At pH levels higher than optimum, when the tail fibers of bacteriophage
T4 are extended, the adsorption rates decrease presumedly because negative
charges on the virus particles and carbon surfaces increase causing greater
repulsive forces between them.
With increasing ionic strength, adsorption increases as like charges on the
virus and carbon particles are masked. At the same time, repulsive forces
between the viral tail fibers and tail sheath are reduced. With increasing ionic
strength, more and more tail fibers adsorb on the tail sheath. Although high
ionic strengths cause a maximum collision frequency between viruses and
carbon particles, the adsorbing tail fibers become less available. These
mechanisms are supported by equilibrium studies at high and low ionic
strengths.
Coulanges, P., Mayoux, A., Brygoo, E. R. (1971). Application of the Soluble
Ultrafilter Membrane Technique to the Study of Viruses in Wastewater of
Tananarive. COMPT REND, 164:1672-3. French.
At Tananarive, filtration of wastewater through soluble alginate mem-
branes yielded viruses in 43% of 67 samples. Thirty-three enteroviruses and a
reovirus were recovered by this method.
Coulon, G., Netter, R. Nejmi, D. (1968). Research on Viruses in Potable
Waters, BULL INST NAT SANTE RECH MED, 23(11-12): 1347-54. French.
Viruses were recovered from one-liter quantities of poliovirus 1-seeded
water by direct pressure filtration (ultrafine Gottingen membranes) or by
vacuum filtration at 4 C (alginate Gottingen membranes). Adsorption onto
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A1(0H)3 followed by filtration through HA Miliipore membranes had the
advantage of speed and simplicity but lacked sensitivity. Recovery of the
virus by adsorption to a Dowex resin and elution in nutrient broth was
inefficient.
In 1967 and 1968, viruses were not recovered from drinking water
tested by two different methods twice a month at four sites around Paris.
A number of water samples did yield viruses of types used experimentally
in the laboratory and were believed to be contaminants.
Culp, R. L. (1971). Virus and Bacteria Removal in Advanced Wastewater
Treatment. PUBLIC WORKS, 102(61:84-8.
The renovation process at Lake Tahoe is reviewed along with data
indicating removal of viruses and bacteria by the process.
De Haas, R. A. (1971). Distribution of Polioviruses in Surinam. ARCH GES
VIRUSFORSCH. 33:72-6.
From the beginning of 1964 to the middle of 1969, polioviruses 1, 2, and
3 circulated continuously among 1- to 5-year-old children. Increased poliovirus
circulation correlated with the appearance of clinical poliomyelitis. Trivalent
oral vaccination programs, administered when increased levels of poliovirus
circulation were observed, only partially reduced circulation of polioviruses,
possibly because insufficient numbers of children were vaccinated.
Dolin, R., Blacklow, N. R., DuPont, H., Format, S., Buscho, R. F-, Kasel, J. A.,
Chanties, R. P., Hornick, R., Chanock, R. M. (1971). Transmission of Acute
Infectious Nonbacterial Gastroenteritis to Volunteers by Oral Administration
of Stool Filtrates, J 1 N F ECT Dl S, 123(3): 307-312.
Acute gastroenteritis was induced in two of three volunteers by oral
administration of a bacteria-free stool filtrate derived from a patient with
naturally occurring acute infectious nonbacterial gastroenteritis. Subsequently,
oral administration of a stool filtrate from one of the sick volunteers induced
gastroenteritis in seven of nine additional men.
The original rectal swab specimen from the naturally occurring case was
passaged three times in human fetal intestinal organ culture: material from the
third passage induced gastroenteritis in one of four volunteers.
A spectrum of clinical responses developed Tanging from a predominantly
diarrhea] disease without vomiting to an illness characterized by repeated
vomiting without accompanying diarrhea.
Geldreich, E. E., Clarke, N. A. The Coliform Test: A Criterion for the Viral
Safety of Water. In "Virus and Water Quality: Occurrence and Control," edited
by V. Snoeyink. Proceedings of the Thirteenth Water Quality Conference,
February 1971. University of Illinois Bulletin, 69 (197 I). 103-13.
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The absence of total coliforms in a treated water is evidence of a
bacteriologically safe supply. Whether coliforms are good indicators for
rickettsia and viruses has not been determined. The observation that certain
enteric viruses are more resistant to chlorine than the indicator bacteria are has
raised questions as to whether drinking water that meets the USPHS drinking
water standard is free from disease-producing levels of virus. Improved methods
for detecting viruses in water, studies on the coliform levels in large volumes of
treated water which are coliform-free by conventional tests, and epidemiological
studies are needed to confirm that the coliform test reliably indicates the
biological safety of water.
Goncharuk, E. I., Grigoryeva, L. V., Bei, T. V., Shulyak, E. V., Korchak,
G. I. (1970). Removal of Enteroviruses and Bacteria from Sewage in
a Circulating Oxidizing Channel. HYG & SAN IT, 35(1):36-41.
Sewage in a circulating oxidizing channel was free of seeded cox-
sackievirus B5 and echovirus 19 in 24 and 48 hours, respectively. Coliphages,
present initially in concentrations of 6,000 to 7,000 PFU/ml, were not
detected 16 hours after seeding. Pathogenic Escherichia coli, present
initially in a concentration of 108/liter, were detected for 18 days.
Coxsackievirus B5 was detected in sludge for three days and echovirus 19
was detected there for five days.
Grigoryeva, L. V., Korchak, G. I., Bey, T. V. (1969). Survival of Bacteria
and Viruses in Sewage Sludges. MIKROBIOL ZH, 31{1l-12>:659-64.
Russian.
Coxsackievirus B5 survived thermophilic fermentation of sludge for
30 days. Enteropathogenic Escherichia coli 0111 survived for 10 days.
Phages survived mesophilic and thermophilic fermentations for four
months and nine days, respectively.
Grushko, Y. M. (1971). Problems of Sanitary Protection of Water Bodies
in the Pages of Foreign Journals. GIG SANIT, 36:73-5. Translation
presently not available. Russian.
Hill, W. F., Jr., Akin, E. W., Benton, W. H. Detection of Viruses in Water:
A Review of Methods and Application. In "Virus and Water Quality:
Occurrence and Control," edited by V. Snoeyink. Proceedings of the
Thirteenth Water Quality Conference, February 1971. University of
Illinois Bulletin, 69 (1971), 17-46: (Also in WATER RES, 5(11):967-95,)
1971.
Mechanisms, procedures, and efficiencies of methods currently under
evaluation for effectiveness in detecting small amounts of viruses in large
volumes of water are reviewed and evaluated for speed, simplicity, and
economy of application.
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The most promising methods are the membrane-adsorption technic,
adsorption to precipitable salts, iron oxide, and polyelectrolytes, aqueous
polymer two-phase separation, and the soluble alginate filter technic. Most
of these methods have good-to-excellent virus recovery efficiencies. Other
methods such as continous-flow ultracentrifugation, forced-flow electro-
phoresis and electroosmosis, and hydroextraction are not as useful yet. The
most promising methods for detecting small amounts of viruses in clean and
finished waters are those that rely on virus adsorption and retention in a
flow-through sampling system. For waters that are moderately or grossly
turbid, the aqueous polymer two-phase separation may be the better
approach.
Hill, W. F., Jr., Akin, E. W., Benton, W. H., Hamblet, F. E. Viral Disinfection
of Estuarine Water by UV. In Proceedings of the National Specialty
Conference on Disinfection, July 1970. ASCE, New York, New York (1971),
223-44: Also in J SAN ENGIN DIV, ASCE, 97:SA5:601-1S, 1971.
Under static conditions in seawater, polioviruses 1, 2, 3, echoviruses
1 and 11, and coxsackievirus A9 were destroyed at about the same rate by
ultraviolet light. Coxsackievirus B1 and reovirus 1 were more resistant,
perhaps because of clumping of the virions. The applied UV dosage was
116 ergs/mm2/second.
In a continuous-flow seawater system, poliovirus 1 was destroyed more
rapidly by a Kelly-Purdy UV unit than in the static system, apparently
because of the higher UV dosage applied in the continuous-flow system.
Johnson, J. D.f Overby, R. Bromine and Bromamine. Disinfection Chemistry.
In Proceedings of the National Specialty Conference on Disinfection, July
1970. ASCE, New York, New York (1971), 37-60.
A pH versus bromine-ammonia mole ratio distribution diagram for
bromine and the bromamines is presented for 10"4 M aqueous bromine
solutions. Ultraviolet absorption spectra were used to identify the principal
bromamine species initially formed, and also to follow the apparent
second and third order decomposition of dibromamine and tribromamine.
In the neutral pH range, tribromamine formed in solutions with mole
ratios of ammonia to bromine less than 2:3; dibromamine occurred at
ratios greater than 2:3. At ammonia to bromine mole ratios of 2:3 in
neutral solution, a breakpoint reaction occurred between di- and tri-
bromamine. Below pH 8 tribromamine was stable. At ratios of ammonia
to bromine of 1:3, pure tribromamine formed and decomposed to nitrogen
and free bromine. At pH values less than 8, the rate of decomposition was
slow in the presence of excess bromine.
Known variations of sporicidal and virucidal efficiencies as a function
of pH, time, and ammonia to bromine ratios correlated with changes in the
chemical species in solution.
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Kinman, 'R. N., Black, A. P., Thomas, W. C., Jr. Status of Water Disinfection
with Iodine. In Proceedings of the National Specialty Conference on
Disinfection, July 1970. ASCE, New York, New York (1971), 11-35.
The qualities and characteristics of iodine as a water disinfectant
are reviewed.
No major difficulties were encountered with iodine during six years
and eight months of iodination of two public water supplies of 25,000
and 100,000 gpd capacities. Iodine had no detrimental effect on general
health or thyroid function and was an effective water disinfectant.
Iodine ($1.30/pound) cannot compete economically with gaseous
chlorine ($0.05/pound) in large water treatment plants at this time. However,
in disinfection applications where hypochlorites are used ($0.70/pound
available chlorine) iodine can compete with chlorine and may be the cheaper
of the two disinfectants depending upon the conditions of use.
Klein, E., Smith, J. K., Morton, F., and Sweet, B. H. (1971). Concentration
of Virus from Water by Osmotic Ultrafiltration II. Mass Transport Aspects.
WATER RES, 5(11): 1067-77.
Mathematical analyses of the physical chemistry aspects of the osmotic
ultrafiltration method for concentrating viruses suspended in water were
utilized for determining optimum experimental conditions. Laboratory
studies showed that the rapidity of concentration depended upon the
starting volume of osmotic and virus solutions, the concentration of the
osmotic solution, the type membrane employed, and the membrane area.
Graphic illustration of the interaction of these parameters and the
derivation of mass transport formulae indicated agreement between exper-
imental and computer predicted results.
Knocke, K.-W., Pittler, H., Hopken, W. (1967). Demonstration of Reovirus
in Sewage. ZBL BAKT ORIG, 203:417-21. German.
Sewage samples, taken from seven towns of Lower Saxony from 1963
to 1965, began to yield large numbers of reoviruses in 1964. In 1965,
130 strains of reovirus were recovered from 382 samples of sewage.
By hemagglutination inhibition tests, 51 isolates were identified as
reovirus 1 and 66 as reovirus 2.
During 1965, 13 reoviruses were recovered from 2,500 specimens
taken from 1,115 patients.
Konowalchuk, J., Speirs, J. I., (1971). A Method for Recovering
Enterovirus from Infected Samples by Adsorption onto Shaken Suspended
Cells. CAN J MICROBIOL, 17:1279-82.
Coxsackievirus B5 in a 5-ml liquid sample was completely recovered
on a single HEP-2 culture in a single 60 x 15 mm culture dish. Adsorption
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was accomplished by constant agitation of the sample with suspended cells
followed by a stationary period to allow m»nolayer formation for plaque
development. Comparative studies indicated that the conventional method
of adsorption onto stationary monolayers would require 50 times as many
dishes and cells for an equal efficiency. The new method may be useful
for recovery of small numbers of viruses from food or water providing
these are nontoxic to cells and contain no bacteria.
Kott, Y. Chlorination Dynamics in Wastewater Effluents. In Proceedings
of the National Specialty Conference on Disinfection, July 1970. ASCE,
New York, New York, (1971), 585-608.
Eight mg/liter chlorine decreased the coliform level of wastewater
effluents to less than 100/100 ml. Fourteen mg/liter did not give better
results.
Eight mg/liter chlorine rapidly destroyed Endamoeba histolytica cysts
in a temperature dependent reaction.
No algae kill was observed in oxidation pond effluents treated with
8 mg/liter of chlorine for two hours.
Kruse, C. W., Hsu, Y. C., Griffiths, A. C., Stringer, R. Halogen Action
on Bacteria, Viruses, and Protozoa. In Proceedings of the National Specialty
Conference on Disinfection, July 1970. ASCE, New York, New York,
(1971), 113-36.
Chlorine, iodine, or bromine, added to water at a low pH, at a
dosage insuring a free residual, would be a universal disinfectant. Less
iodine than chlorine or bromine would be needed to provide a free
residual.
The cysticidal properties of free iodine, bromine, and chlorine in water
of low pH is about the same on a mg/liter basis. Chlorine and, to a lesser
extent, bromine fail as cysticides in water containing nitrogenous substances,
although some amines are fair disinfectants.
The virucidal properties of free halogens are similar. Since neither
infective RNA nor DNA is destroyed by iodine, the optimum pH for
viral iodination is greater than 6. Free bromine and chlorine are more
virucidal at lower pH levels.
On a weight basis, iodine kills more viruses in a wide range of natural
waters than either chlorine or bromine because amines are not formed with
iodine.
Kruse, C. W., Olivieri, V. P., Kawata, K. The Enhancement of Viral Inactivation
of Halogens. In "Virus and Water Quality: Occurrence and Control," edited by
V. Snoeyink. Proceedings of the Thirteenth Water Quality Conference,
February 1971. University of Illinois Bulletin, 69(1971), 197-209.
Free halogens are virucidal in the pH range of natural waters. To dis-
infect water supplies, chlorine should be applied beyond the breakpoint to
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inactivate viruses. Longer contact time gives better disinfection; thus, pre-
chlorination is frequently a desirable water treatment practice. Flash mixing is
of great value when combined residuals are used because free chlorine does
exist for a short time after mixing. Disinfection can be accomplished even in
this short time.
Lautier, F., Wilenski, A., Lavillaureix, J. (1971). A Method for Demonstrating
Viruses in Polluted Waters. PATH MICROBIOL, 37:99-104. French.
One-hundred ml of filtered sewage was mixed with Al(OH)3} and the
precipitate was trapped on a 0.45 jum membrane filter. Viruses were eluted
with culture medium containing protein. Viruses were recovered from most of
19 sewage samples tested. The method was superior to a resin method tested
concurrently.
Layton, R. F., Kinman, R. N. New Analytical Method For Ozone. In Pro-
ceedings of the National Specialty Conference on Disinfection, July 1970.
ASCE, New York, New York, (1971), 285-305.
A colorimetric method for detecting ozone in air and water is described.
The method is based on the oxidation of leuco crystal violet via a hydride
transfer mechanism.
Leclerc, H. (1971). Pathogens in Residual Waters: Evolution in the Course of
Purification Treatments. TSM - L'EAU, 66(11):389-400. French.
Viruses were rarely recovered from raw sewage, and could not be re-
covered from experimentally seeded sludge treated with lime and iron per-
chloride and vacuum filtered. Viruses were recovered from seeded sludge
flocced with an organic polyelectrolyte and pressure filtered. Results of
studies on the removal of other microorganisms by treatment processes are
presented.
Leogrande, G., landolo, E. (1969). Effect of Seawater on Survival of Virulent
and Attenuated Polioviruses. IG MOD, 62:705-14. Italian.
Survival of six poliovirus strains (three virulent and three attenuated) in
seawater, autoclaved (120 C) seawater, and in Hanks' balanced salt solution
were studied. In ten experiments, highly variable results were obtained, but
test conditions were highly variable because of the physical, chemical, and
microbiological (bacterial and protozoan) variations in the seawater. However,
viruses were inactivated in seawater very slowly, at a rate apparently not much
different than in Hanks'.
The authors concluded that seawater has no unusual virus-inactivating
capacity.
12
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Liu, O. C. Viral Pollution and Depuration of Shellfish. In Proceedings of the
National Specialty Conference on Disinfection, July 1970. ASCE, New York,
New York (1971), 397-428.
Viral pollution and depuration of shellfish are reviewed.
In the last 15 years, nine outbreaks of infectious hepatitis have been
associated with various species of shellfish. Recently, sporadic cases of
hepatitis have been associated with raw and inadequately cooked shellfish.
Field investigations over the northeastern United States showed a variety
of human enteric viruses in American oysters and in their growing waters.
Viruses localize in the digestive diverticula of shellfish. The process of
contamination is dynamic. The viruses are retained in the diverticula when
water temperatures are below a certain level. Thus, the shellfish serve as a
reservoir for virus dissemination.
Shellfish can self-cleanse themselves when subjected to optimal hydro-
graphical conditions.
Liu, O. C., Brashear, D. A., Seraichekas, H. R., Barnick, J. A., Metcalf, T. G.
(1971). Virus in Water. L A Preliminary Study on a Flow-Through Gauze
Sampler for Recovering Virus from Witters. APPL MICROBIOL 21{3):405-10.
The efficiency of a flow-through gauze sampler for recovering viruses
from both fresh water and seawater was evaluated. An attenuated strain of
poliovirus 1 was seeded into water in the preliminary laboratory studies.
The amounts of poliovirus 1 adsorbed by the gauze pads from tap water
were very small, about 2% of the total number of virus particles flowing through
the sampler. Virus recoveries from seawater ranged from 15 to 19%. Addition
of NaCl to tap water increased virus recovery to 47%,
Viral elution from the pads was greatest in pH 8 to 9 buffer containing
serum. Repeated elutions from the pads increased viral recovery. The first
eluate contained approximately 50% of the adsorbed virus.
Liu, 0. C., Seraichekas, H. R., Akin, E. W., Brashear, D. A., Katz, E. L., Hill,
W. J., Jr, Relative-Resistance of Twenty Human Enteric Viruses to Free
Chlorine in Potomac Water. In "Virus and Water Quality: Occurrence and
Control," edited by V. Snoeyink. Proceedings of the Thirteenth Water Quality
Conference, February 1971, University of Illinois Bulletin 69 (1971), 171-95.
A wide range of resistance was observed among polioviruses 1, 2, and 3,
five coxsackieviruses, six echoviruses, three adenoviruses, and three reoviruses
subjected to chlorination in partially treated Potomac River water.
A reovirus that required 2.7 minutes for 99.99% devitalization by 0.5
mg/liter of free chlorine was the least resistant virus tested, and a poliovirus that
required 40 minutes for 99.99% devitalization was the most resistant. These
findings are in general agreement with those reported in the literature when
comparable information is available.
Naturally occurring viruses may be much more resistant to chlorination
than laboratory strains,
13
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Malaviya, A. N., Sama, S. K., Ramachandran, K.f Gandhi, P. C., Tandon, B. N.
(1971). Australia/SH Antigen and Immunoglobulins in an Epidemic of Viral
Hepatitis. AM J TROP MED HYG, 20(3):502-5.
In a waterborne epidemic of viral hepatitis, none of 97 persons with
hepatitis showed evidence of infection with Australia antigen. In 26 persons
randomly selected from those with hepatitis, IgG and IgM levels were
significantly elevated.
Manwaring, J. F.f Chaudhuri, M., Engelbrecht, R. S. (1971). Removal of
Viruses by Coagulation and Flocculation. J AMER WATER WORKS ASSN,
63(51:298-300.
In laboratory studies, coagulation of coliphage MS2 in water with
FeCl3 was as effective as coagulation with A12(S04)3. The optimum con-
centration of FeCl3 was 50 to 60 mg/liter. Calcium and magnesium concentra-
tions up to 50 mg/liter each did not affect the efficiency of coagulation. Virus
and turbidity removal paralleled each other. Removal efficiency increased as the
pH decreased over a range from 8 to 5, and reached a maximum of 99.5%.
The addition to the water of 20% wastewater reduced the efficiency of
virus removal to less than 70%. Wastewater may have interfered more with virus
removal from FeCl3-coagulated water than with virus removal from water
coagulated with A12(S04)3.
Virus was recovered from the settled ferric-sludge indicating that sludge
disposal must be undertaken with care.
Mehta, N. R., Acharya, P. T. (1969). Study of an Outbreak of Infective
Hepatitis in the Hospital Staff Quarters, Baroda. I. Epidemiological Aspects.
IND J MED RES, 57(8): 1437-48.
A waterborne outbreak of infectious hepatitis limited to young adult
males occurred in families of the staff of a hospital and medical college in
Baroda in February, March, and April, 1966. Forty-two cases were reported.
Superchlorination of water and administration of gamma globulin were
instituted to control the outbreak.
Melnick, J. L. Detection of Virus Spread by the Water Route. In "Virus and
Water Quality: Occurrence and Control," edited by V. Snoeyink. Proceedings
of the Thirteenth Water Quality Conference, February 1971. University of
Illinois Bulletin, 69 (1971), 114-24.
Viruses occur in water, and waterborne outbreaks of viral diseases have
been well-documented. If a virus infection has a short and uniform incubation
period, and produces a characteristic, easily recognizable disease, carriage of the
virus by water routes can be traced with a fair degree of accuracy.
In contrast to diseases whose spread can be traced in this fashion, the
characteristics of many viral diseases are such that their transmission by water
14
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is very difficult to recognize. Viruses whose spread is difficult to trace include
those which produce clinically observable illness in only a small fraction of the
persons who become infected, those viruses which produce diseases with
widely variable incubation periods, and those viruses which are readily spread
by direct human contact.
Even though water transmission is often difficult to trace, the presence of
viruses in sewage effluent, in streams, and in other waters cannot be viewed
with equanimity. New methods capable of detecting viruses in large fluid
volumes are available, and the time may have arrived for setting standards of
permissible virus content of waters. A maximum is proposed of one detectable
infectious virus unit/10 gallons of water intended for swimming and bathing
purposes, and of no more than one infectious virus unit/100 gallons of
drinking water.
Metcalf, T. G. Biologic Parameters in Water Transmission of Viruses. In "Virus
and Water Quality: Occurrence and Control," edited by V. Snoeyink. Pro-
ceedings of the Thirteenth Water Quality Conference, February 1971. Uni-
versity of Illinois Bulletin, 69 (1971), 1-16.
The properties of animal viruses important to man and transmitted by
water are discussed. The salient features of these viruses which set them apart
from bacteria and which must be understood to understand the problems of
viruses in water are described.
Mitchell, R. (1971). Destruction of Bacteria and Viruses in Seawater. J SAN
ENGR DIV, ASCE, 97:SA4:425-32.
The rate of inactivation of enteric bacteria in seawater was proportional
to the quantity of marine microflora present. Continued disposal of enteric
microorganisms into the sea results in the development of a specific antagonistic
microflora.
Three groups of microorganisms have been associated with the destruc-
tion of coliforms in the sea. They are bacteria which enzymatically lyse the cell
walls of enteric bacteria, the obligately parasitic Bdellovibrio, and a group of
amebae that attack and consume bacterial cells. The amebae appear to be the
most effective of the three groups.
Viruses in seawater apparently are destroyed by native marine microflora
in the same way enteric bacteria are destroyed, but the antiviral microorganisms
have not been identified. A chemical component of the seawater, probably a
heavy metal, is also involved in the destruction of viruses. Shellfish-associated
infectious hepatitis indicates, however, that viruses in the sea may survive
inactivation. Viruses are protected from inactivation in the sea for long periods
by adsorption to living and dead bacterial cells and to cell debris.
Moro, S. (1970). Technics for the Virological Analyses of Water and Sewage.
G BATTERIOL VIROL IMMUN, 63(9-10) 467-84. Italian.
Some methods for detecting viruses in water are reviewed.
15
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Morris, J. C. Disinfectant Chemistry and Biocidal Activities. In Proceedings of
the National Specialty Conference on Disinfection, July 1970. ASCE, New
York, New York (1971), 609-33.
To understand the kinetics of disinfecting action, the germicidal process
must be viewed as a mutual interaction between each specific molecular form
of the chemical agent and the microbes (considered as biochemical units). For
the chemical agent, the positions of chemical equilibria and the germicidal
potency of each of the active species must be known. For the organism, the
physical and chemical nature of its areas of interaction with the chemical
agent, and its susceptibility changes in different solutions must be known.
Morse, L. J., Bryan, J. A., Chang, L. W., Hurley, J. P., Murphy, J. F., O'Brien,
T. F. (1970). Holy Cross Football Team Hepatitis Outbreak. ANTIMICROB
AGENTS CHEMOTHER, 10:30-2.
An outbreak of infectious hepatitis occurred at Holy Cross College
among players and coaches of the football team during September and
October, 1969. Ninety of the 97 persons exposed were infected. Thirty-two
experienced icteric disease, 22 were anicteric but symptomatic, and 36
asymptomatic players experienced elevated serum glutamic pyruvic trans-
aminase values (>100 units).
The source of infection appeared to be a water supply contaminated by
back siphonage into an open, flooded, below-ground irrigation faucet when a
fire department placed heavy demand on the supply several miles away. The
faucet area had been flooded and contaminated by playing children who had
just recovered from infectious hepatitis.
Nestor, I. (1970). Methods for Isolation of Viruses from Water, STUD CERCET
INFRAMICROBIOL, 21(4):345-57. Translation presently not available.
Rumanian.
Nestor, I., Costin, L. (1971). The Removal of Coxsackievirus from Water
by Sand Obtained from the Rapid Sand Filters of Water Plants. HYG
EPIDEM MICROBIOL IMMUN, 15: 129-36.
Fresh dry sand removed viruses much less efficiently than wet sand.
Both experimentally and in field studies, sand from functioning filters more
effectively removed viruses than other sands.
Nesvold, H. P. (1970). Infectious Hepatitis, Hygienic Problems Related to
an Epidemic in a Hamlet in Trysil. TIDS NOR LAEGEFOREN, 90(10):
1882-8. Translation presently not available. Norwegian.
Nitzkin, J. L., Henry, M. H. (1971). Infectious Hepatitis in Logan County,
Kentucky, A Probable Common Source Outbreak. J KY MED ASSOC,
69(5): 349-53.
16
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Ninety cases of viral hepatitis were reported in a six-county area of
Western Kentucky during September, October, and November, 1967. Seventy-
six of the 90 patients were residents of Logan County, and 64 were between
15 and 24 years of age. Soft drinks prepared from unchlorinated well water
at a public eating place in Logan County appeared to be the common
source of 74 cases. Five secondary cases occurred.
Nupen, E. M. (1970). The Isolation of Viruses from Sewage and Treated
Sewage Effluents. WATER POLLUT CONTRL(So. Africa), 69:430-4.
The occurrence of viruses in sewage and effluents, their removal by
treatment processes and disinfection, and methods for concentrating and
detecting them are reviewed.
Olifson, L. E., Baltenko, E. N., Bukharin, O. V., Pozhar, V. N., Turovets,
G. L. (1970). Utilization of Industrial and Agricultural Wastewaters. GIG
SANIT, 35<12):92-4. Translation presently not available. Russian.
Olivieri, V. P., Donovan, T. K., Kawata, K. Inactivation of Virus in Sewage.
In Proceedings of the National Specialty Conference on Disinfection, July
1970. ASCE, New York, New York (1971), 365-84: Also in J SAN ENGIN
DIV, ASCE, 97:SA5:661-73, 1971.
Data are presented that strongly support the hypothesis that free
chlorine is required for significant inactivation of viruses in sewage. The
occurrence of short-lived virucidal intermediates and a long-lived chlorine-
virus complex does not appear significant. The short-lived presence in sewage
of free chlorine, as determined by a modification of the Black and Whittle
procedure, appears likely when coupled with the results of dynamic tests.
Palfi, A. B., Simon, M., Schulek, E. (1970). Virological Examination of Urban
Sewage from Budapest. II. Virus Isolations. EGESZSEGTUDOMANY, 14:
127-32. Hungarian.
In the six-week periods following vaccination programs with monovalent
live poliovirus vaccines, polioviruses were continuously recovered.
In August and September, coxsackieviruses and echoviruses, primarily
coxsackievirus B4 and echovirus 7, were recovered. Reoviruses were recovered
in the later period of the study.
Trickling-filter and activated-sludge plants did not effectively reduce
virus concentrations.
Pittler, H., Hopken, W., Knocke, K.-W. (1967). Enteroviruses and Adenoviruses
in Sewage: Demonstration and Epidemiological Study 1963-1965. Z BAKT
ORIG, 204:33-48. German.
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From 1963 to 1965, viruses were detected in 55% of 381 sewage samples
collected from ten towns of Lower Saxony. Samples were concentrated at
first by ultracentrifugation and later by precipitation with polyphosphoric
acid.
Polioviruses, coxsackieviruses, echoviruses and adenoviruses were found.
Most of the enteroviruses recovered from patients during this period were also
found in sewage. There were no viruses in the sewage that were not also
recovered from patients at about the same time. Most viruses were recovered
during the summer and autumn.
Primavesi, C. A. (1969). Modern Problems of Water Management and Water
Distribution. I. GELSENKIRCHEN, 4:195-213. German.
The occurrence and survival of viruses in sewage and river waters are
reviewed.
Primavesi, C. A. (1971). Virology and Its Importance for Water Hygiene.
OEFF GESUNDHEITSWES, 33(6):330-6. German.
The occurrence of viruses in sewage and sludge are reviewed.
Sewage containing fecal matter nearly always contains enteroviruses.
Coxsackieviruses and echoviruses occur in sewage during the summer and
autumn months. Polioviruses occur in sewage following vaccination programs.
The epidemiology of enterovirus infections in the community is re-
flected in the sewage. Enteroviruses are found in the influents and effluents
of sewage treatment plants. Treatment processes do not substantially reduce
the virus content of sewage. Enteroviruses are present also in sewage-
containing canals. These canals contain viruses in the winter, summer, and
autumn.
Enteroviruses are also present in treatment sludges even after months of
storage. This is important in the agricultural utilization of sludges.
Reddick, R. A., Aycock, E. K., (1971). Water - A Vehicle of Disease
Dissemination. J SC MED ASSOC, 67{2): 56-60.
Viral diseases transmitted through contaminated water are reviewed
along with bacterial and protozoan diseases transmitted by this route.
Ringertz, O. (1971). Some aspects of the Epidemiology of Hepatitis in
Sweden. POSTGRAD MED J, 47(7):465-72.
An outbreak of infectious hepatitis associated with the consumption of
oysters occurred in 1955.
Subsequently, an increasing incidence of hepatitis was noted among
cross country runners who apparently crossinfected each other directly and
indirectly.
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Since 1964, hepatitis associated with haemodtalysis and with narcotic
addiction has become common.
The changes in the epidemiological pattern of infectious hepatitis during
the last 20 years are discussed.
Rubenstein, S. H., Orbach, H. G.,, Shuber, N., King, E., Zackler, J. (1971).
Freeze Concentration of Viral Agents from Large Volumes of Water. J AMER
WATER WORKS ASSN, 63(51:301-2.
Poliovirus 1 and Newcastle disease virus were freeze-concentrated from
five-liter quantities of distilled water in a Virtis Freeze Concentrator. The
process took eleven hours. Seventy to 125 ml of water remained unfrozen at
the end of the process. One-hundred fold concentration of both viruses was
noted when high concentrations of viruses had been added to the test waters.
The efficiency of virus recovery was not reported.
Scarpino, P. V. Bacterial and Viral Analysis of Water and Wastewater. In
"Water and Water Pollution HandbookChapter 13, edited by L. Ciaccio.
Marcel-Dekker, New York, New York, (19711, 639-761.
This chapter includes a comprehensive literature review on viruses in the
water environment. Although its major emphasis is on methodology, the review
covers occurrence, importance, detection, and quantitation of viruses in
waste and other waters.
The major segment of the chapter deals with bacteria, primarily those of
sanitary significance, in the water environment and analyses related to them.
Schafer, E. (1971), Quantitative Recovery of Poliovirus 2 in Surface Waters.
GAS WATER ASSN/SEWAGE, 112(21:109-13. German.
Ultrafiltration through lanthanium-aluminum-alginate filters, adsorption
to 0.45 jum cellulose nitrate filters (after prior filtration through an ion
exchange resin), two-phase separation, and FeCl3 floculation were compared
for their efficiencies in recovering poliovirus 2 from seeded surface waters.
In numerous experiments, all of the methods had efficiencies that
averaged about 60 to 70% except the membrane filter procedure which
recovered less than 30% of the virus. Combining FeCl3 floculation with two-
phase separation yielded less than 30% of the virus, but reduced the final
volume to a few ml.
Schneweis, K. E., Stifter, G. (1971). Recovery on Membrane Filters of Entero-
viruses in Water,: ZBL BAKT ORIG, 216: J 28-39. German.
A poliovirus, seeded into one-liter quantities of phosphate-buffered tap
water (pH 7.4), adsorbed onto collodion membranes with an average pore
diameter of 200 nm and was eluted by passing three ml of tissue culture
medium containing 2% calf serum and 1% gelatin through the membranes.
19
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Eighty-four percent of the virus was recovered. One and three-tenths infectious
units of poliovirus was detected in 900 ml of the water.
The method was faster, simpler, and as efficient as the alginate filter
technic. Bacteria were retained on the filter, and the eluted material produced
no toxicity to tissue cultures. Extensive studies with coliphage T3 also yielded
good results.
With sewage, 300 ml quantities of gauze pad elutes were centrifuged
at 8,000 to 10,000 rpm for one hour, filtered successively through filterpaper
(Schleicher and Schull 595-1/2), through collodion membranes with approxi-
mate pore diameters of 3,000 (with adsorbing pad), 1200, 800, 450 and 200
nm, and through a Dowex column. Magnesium chloride was then added and
the viruses were adsorbed onto and eluted from a collodion membrane as
described for tap water. Virus recoveries were lower than from tap water, but
the technic was simpler and apparently more sensitive than the alginate filter
method. Enteroviruses were detected in 41 of 43 samples tested.
Scutt, J. E. (1971). Virus Retention by Membrane Filters. WATER RES, 5(5):
183-5.
Twenty ml amounts of phosphate-buffered saline containing about 338
PFU/ml reovirus or 216 PFU/ml poliovirus 1 were filtered through 0.45 fxm
cellulose nitrate membranes at pressures ranging from about 0.023 to 3.54 mm
S"1.
Retention of poliovirus 1 by filters decreased with increasing filtration
rates. Reovirus 1 was completely retained by these membranes at all rates
examined. The poliovirus results suggest that membrane filtration may be an
unsatisfactory method for concentrating viruses from large volumes of water.
Shane, M. S. (1971). Distribution of Blue-Green Algal Viruses in Various
Types of Natural Waters. WATER RES, 5(9):711-16.
LPP blue-green algal viruses were recovered from rivers, streams, lakes,
farm and residential ponds, oxidation lagoons and industrial storage waters.
These viruses were widely distributed in nature and stable throughout the
year in three oxidation ponds examined in the Delaware-Maryland area.
Shuvat, H. I., Thompson, A., Fattal, B., Cymbalista, S., Wiener, Y. Natural
Virus Inactivation Processes in Sea Water. In Proceedings of the National
Specialty Conference on Disinfection, July 1970. ASCE New York, New
York (1971), 429-52: Also in J SAN ENGIN DIV, ASCE, 97: SA5:587-
600, 1971.
Enteroviruses were more resistant to inactivation in the marine environ-
ment than coliform organisms which have been the classical pollution indicator.
Enteroviruses were often detected in a sewage-polluted coastal area and
opposite a bathing beach where coliform counts were low.
20
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Seawater samples from sewage polluted areas of the Mediterranean
and Red Seas had higher marine antiviral activity (MAVA) than samples
of unpolluted seawater.
Heat and ether treated samples lost their antiviral activity, as did
samples passed through membrane filters. All antiviral seawater samples
contained viable marine bacteria with an average density of 103/ml.
Active antiviral factors could not be demonstrated in seawater in the
absence of marine bacteria. Whether the antiviral activity requires the presence
of the bacteria or is associated with an agent produced by the bacteria has not
been determined.
An effect similar to MAVA was detected in eight pure cultures of
marine bacteria. The morphological and biochemical characteristics of a
number of these organisms differed widely. On multiple sub-cultivation
their antiviral activity was lost. MAVA has been observed in the Atlantic
Ocean, North Sea, Baltic Sea, Mediterranean Sea and Red Sea.
Simon, M., PSIfi, A. B. (1970). Virological Examination of Urban Sewage
from Budapest /. Concentration of Virus by Adsorption to Calcium Phosphate.
EGESZSEGTUDOMANY 14:13-18. Hungarian.
Viruses in sewage were adsorbed onto CaHP04. A purified suspension
for inoculation was prepared by subjecting the adsorbed virus first to
phosphate buffer of high molarity and then to buffer of low molarity.
Sobsey, M. D., Cooper, R. C. Laboratory Studies on the Survival of Polio-
virus in Algal-Bacterial Wastewater Treatment Systems. In "Virus and Water
Quality: Occurrence and Control," edited by V. Snoeyink. Proceedings of the
Thirteenth Water Quality Conference, February 1971. University of Illinois
Bulletin, 69 (1971), 137-47.
In laboratory studies, considerable antipoliovirus 1 activity was observed
in stabilization pond water and in algae-bacteria cultures. Bacterial cultures
alone showed less virus-inactivating capacity, and a pure culture of the alga
Scenedesmus quadricauda had none. Antiviral activity appeared to be associ-
ated with either biological activity or heat labile factors, but the specific
causes were not identified.
Sollo, F. W., Jr., McGurk, F. F., and Larson, T. E. Status of Methods for
Halogen Determination (Br-Cl). In Proceedings of the National Specialty
Conference on Disinfection, July 1970. ASCE, New York, New York (1971),
245-67.
Analytical technics for determining halogens and some of their com-
pounds are reviewed and compared.
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Sorber, C. A., Sagik, B. P., Malina, J. F., Jr. (1971). Monitoring of Low-
Level Virus in Natural Waters. APPL MICROBIOL, 22(3>:334-8.
The effectiveness of the insoluble polyelectrolyte technic (PE 60) for
concentrating cotiphage T2 was a constant 20% over a range of virus levels
from 103 to 104 PFU/ml. The efficiency of the method was dependent upon
pH during the concentration phase.
Sproul, O. J. Recent Research Results on Virus Inactivation by Water Treat-
ment Processes. In "Virus and Water Quality: Occurrence and Control,"
edited by V. Snoeyink, Proceedings of the Thirteenth Water Quality Con-
ference, February 1971. University of Illinois Bulletin, 69 (1971), 159-69.
Research data collected within the past five years on virus inactivation
by water softening and alkaline pH conditions, coagulation, filtration, and
adsorption are reviewed. Research efforts presently under way are also
reviewed.
Sproul, O. J., Thorup, R. T., Wentworth, D. F., At well, J. S. Salt and Virus
Inactivation by Chlorine and High pH. In Proceedings of the National
Specialty Conference on Disinfection, July 1970. ASCE, New York, New
York (1971), 385-96.
Increased divalent cation concentration increased the rate of inactivation
of T2 coliphage by chlorination.
The pH required to produce significant inactivation of poliovirus 1 was
higher than that previously reported. No inactivation occurred after 90
minutes at pH 10.5 produced with NaOH and Ca(OH)2, or at 11.5
produced with KOH.
Sullivan, R., Fassolitis, A. C., Larkin, E. P., Read, R. B., Jr., Peeler, J. T.
(1971). Inactivation of Thirty Viruses bv Gamma Radiation. APPL MICRO-
BIOL, 22(1):6l-5.
The weighted decimal reduction values (D values) of 30 viruses sus-
pended in Eagle's minimum essentia! medium ranged from 0.39 to 0.53
Mrads. Only one-third of this radiation dose was required to achieve equal
inactivation rates in distilled water. The destruction rate curves wore those of
a first-order reaction.
Sweet, B. H,, Klein, E., Ellender, R„ Morton, F. C., Smith, J. K. Concentra-
tion of Viruses by Osmotic Ultrafiltration: A Preliminary Report on the
Development of a Model System In "Virus and Water Quality: Occurrence
and Control," edited by V. Snoeyink. Proceedings of the Thirteenth Water
Quality Conference, February 1971. University of Illinois Bulletin, 69(1971),
51-8.
The basic physical features of an osmotic ultrafiltration unit and the
equipment and solutions employed in experimental studies to concentrate

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poliovirus 1 suspended in distilled water are described. The mainstays of the
system are the asymmetric cellulose acetate membranes which separate the
osmotic and virus compartments.
The system reduced 20 liters of sterile water to 50 ml in 5-1/4 hours.
Dehydration of virus-inoculated water allowed 100% recovery of virus.
Sweet, B. H., McHale, J. S., Hardy, K. J., Morton, F., Smith, J. K. and Klein,
E. (1971). Concentration of Virus from Water by Osmotic Ultrafiltration-],
Biological Aspects, WATER RES, 5(101:823-9.
Distilled water seeded with poliovirus 1 was dehydrated by osmotic
ultrafiltration in a two compartment system utilizing distilled water as the
virus suspending medium and saturated magnesium sulfate, sucrose, or both
solutions as the osmotic driving force. The compartments were separated by
asymmetric cellulose acetate, reverse osmosis-type membranes.
A minimum virus inoculum of 1.3 PFU/ml could be detected in a 4-liter
starting volume. Virus recoveries of 90% or greater were achieved.
Telniceanu, A. (1971). Epidemiologic Relationships in Viral Hepatitis. Hous-
ing Conditions and Water Transmission. STUD CERCET INFRA MICROBIOL,
22:113-22. Rumanian.
The relationships between living conditions (housing, district, drinking
water supplies) and the epidemiologic character of hepatitis morbidity were
evaluated in four areas of Bucharest during an epidemic in 1969 and 1970.
Telniceanu, A. (1971). The Epidemic Evolution of Infectious Hepatitis Under
Conditions of Intermittent Water Transmission in Urban Areas. STUD CERCET
INFRA MICROBIOL, 22:123-33. Rumanian.
The relationship between infectious hepatitis incidence and chlorination
of drinking water was investigated. Forty-nine periods, where chlorine was
absent, were correlated with infectious hepatitis occurring 31 to 42 days later.
Vajdic, A. H. (1970). The Isolation of Virus (Bacteriophage) from Water and
Wastewater by the Millipore Filter Method WATER MANAGEMENT IN
ONTARIO (Ontario Water Resources Commission), 37:1-22.
A phage of Escherichia coli B, in quantities from two to more than 500
MPN, was seeded into volumes of water up to 20 liters and recovered by
adsorption onto 0.45 jum cellulose nitrate membrane filters and eluted with
3% beef extract. Recovery efficiencies ranged from 0 to 100%. Reovirus
recoveries were greatest from distilled water containing calcium. Lesser
recovery efficiencies were experienced with dechlorinated tap water, well
water, and surface water. The phage were also recovered from sewage with
this method.
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VaneckovA, N., Koza, J., Mikesova, V. (1971). Systematic Follow-up Study
of the Circulation of Polioviruses and Other Enteroviruses in Wastewater. CS
EPIDEM, 20(1): 18-26. Czechoslovakia!!.
From November 1962 to October 1966, in ten different regions, 96
viruses were recovered from 229 sewage samples including 44 polioviruses, 20
echoviruses, 8 coxsackieviruses, and 24 untypable viruses. Wild-type poliovirus
strains were also recovered from wastewater in two regions.
Verlinde, J. D.t Wilterdink, J. B. (1970). Neurovirulence of Polioviruses.
ANTONIE VAN LEEUWENHOEK 36:587.
From 1964 to 1968, polioviruses were recovered from stools of healthy
children in day nurseries and from samples taken from sewage treatment
plants.
Five strains each of polioviruses 1, 2, and 3, recovered during the five
years of the study, were selected at random and examined for neurovirulence
by intraspinal inoculation of cynomolgus monkeys. The degree of neuro-
virulence was measured by the amount of virus that produced paralysis or
characteristic histological lesions.
The neurovirulence of the strains examined ranged from highly neuro-
virulent to highly attenuated. Most strains were intermediate.
Verlinde, J. D., Wilterdink, J. B. (1970). Neurovirulence of Polioviruses
Recovered from Sewage and Stools of Healthy Children. II. Neurovirulence
of Strains of Types 1 and 3. ARCH GES VIRUSFORSCH, 32:311-17.
The monkey neurovirulence of five strains of poliovirus 1 and five
strains of poliovirus 3, recovered from stools of healthy children in day
nurseries and from sewage samples taken over a five-year period (1964-1968),
was examined. Neurovirulence ranged between the extremes of highly neuro-
virulent and completely attenuated. The majority of strains possessed an
intermediate neurovirulence.
Wallis, C. Progress in the Development of an Apparatus for Concentration of
Viruses from Large Volumes of Water. In "Virus and Water Quality:
Occurrence and Control," edited by V. Snoeyink. Proceedings of the Thirteenth
Water Quality Conference, February 1971. University of Illinois Bulletin, 69
(1971), 47-50.
Poliovirus 1 added to tap water was efficiently recovered by the follow-
ing procedure. The water was passed through clarifying filters with porosities
of lOjum, 5/j.m, 3jim, and ljum to remove particulate matter, and then through
a ljum cotton textile filter to electrostatically remove ferric compounds. The
water was then passed through a special amberlite resin to remove other
insoluble salts and fine silts not removed by the textile filter, and the clean
water containing the virus was driven through a virus adsorbent, either a
cellulose nitrate membrane or the insoluble polyelectrolyte PE 60. MgCl2
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was added to enhance the adsorption of virus on cellulose nitrate membranes
when these were used, and C02 was added to concentrate the virus on the
polyelectrolyte when this was used. Water could be processed at the rate of
300 gph. Virus recovery efficiencies ranged from 61 to 84%.
Wallis, C., Melnick, J. L., Fields, J. E. (1970). Detection of Viruses in Large
Volume of Natural Waters by Concentration on Insoluble Poly electrolytes.
WATER RES, 4(12):787-96.
Poliovirus 1, added to large volumes of tap water, could be recovered by
adsorbing it onto thin layers of an insoluble polyelectrolyte (a cross linked
copolymer of isobutylene maleic anhydride), and subsequently concentrating
the virus into small volumes of elutant for assay. Virus contained in 25, 50, 75
and 100 gallons of water was recovered with efficiencies ranging from 60 to
80%. Poliovirus 1, added to a 17,000 gallon swimming pool, was recovered
with an efficiency of about 40% by passing 300 gallons of the pool water
through a thin 3 mm polyelectrolyte layer 293 mm in diameter, and sub-
sequently eluting the virus from the layer.
Wallis, C., Melnick, J. L., Fields, J. E. (1971). Concentration and Purification
of Viruses by Adsorption to and Elution from Insoluble Poly electrolytes.
APPL MICROBIOL, 21 (4): 703-709,
Acid-resistant, nonenveloped viruses of the enterovirus, reovirus, and
adenovirus groups concentrated on PE 60, an insoluble cross-linked poly-
electrolyte based on isobutylene maleic anhydride. NaOH hydrolysis of PE 60
increased its capacity to adsorb viruses. Maximal virus adsorption occurred at
pH levels between pH 3.0 and 4.5, and the adsorbed viruses were readily eluted
at pH 8 to 9.
Volumes of cell culture fluids up to one liter, containing 108 PFU of
virus/ml, were tested. Virus recoveries exceeded 60%.
Whittle, G. P. (1970). Recent Advances in Determining Free Chlorine, In Pro-
ceedings of the National Specialty Conference on Disinfection, July 1970.
ASCE, New York, New York (1971), 269-283.
The leuco-crystal violet (LCV) method apparently measures completely
unbound, free chlorine present at any instant in time. This specificity
apparently stems from inhibition of hydrolysis of N-chlor compounds brought
about by mercuric chloride, a reagent in the LCV method.
Application of the LCV method and other methods to solutions of
chlorine and cyanurate failed to show significant concentrations of com-
bined chlorine. An explanation of the stabilization and reduced biocidal
activity of solutions of chlorine and cyanurate based on ultraviolet energy
absorption by the cyanurate molecule was tendered.
Zdrazilek, J., Zdcek, K., Chvapil, J., Mikesova, V., Pokorna, L.t Tomanova, V.,
Trauc, J., Vrabkova, J. (1971). Virological Surveys for Enteroviruses in
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Wastewater. /. Incidence of Polioviruses in Prague at the End of 1968 and in
1969\ CS EPIDEMIOL MICROBIOL IMMUN, 20(3):67-72. Czechoslovakian.
In 1968 and 1969, 180 gauze pad samples were collected from 10
points in the main Prague sewers. Polioviruses were recovered from 21 samples,
other enteroviruses from 71 samples, and mixtures of polioviruses and other
enteroviruses from two samples. No enteroviruses were recovered from 86
samples.
With the exception of one strain of poliovirus 3, recovered from sewage
in February 1968, polioviruses were recovered from samples collected only
from April to July, a period following mass vaccination of newborn children
with live poliovirus vaccine.
Polioviruses apparently did not circulate in the population during the
winter months, the period during which they occurred in greatest numbers
before introduction of vaccination.
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