VIRUSES IN  WASTE,
W RENOVATED,
AND OTHER WATERS
          1972
  LITERATURE ABSTRACTS


   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. Dianne White
                1972

           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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                                1972

  VIRUSES IN WASTE. RENOVATED, AND OTHER WATERS
Ahmed, T.  M. A.  (1972).  Effect of Ultraviolet Irradiation  on Chlorine
Residuals in Polluted Waters. BIOCHEM J, 128(1): 39.

     More than 50 samples of river water, polluted with sewage and industrial
wastes containing high  concentrations of suspended  solids and dissolved
impurities, were  chlorinated and exposed  to ultraviolet  irradiation  for six
hours.  About  60% of the chlorine dissipated in  the  first two hours. Less
chlorine was lost at  the greater depths. A reduction in pH level  accompanied
the chlorine loss.
Andriashvili, I. A.,  Tikhonenko, T. I., Gushchin, B. V. (1971).  Isolation,
Concentration, Purification and Biological Properties of Enteric Phages. VOPR
VIRUSOL, 16(9-10): 532-4. Russian.

     Five phages of the Fl series were recovered from sewage on Escherichia
coli K12F+. The phages were serologically unrelated to phages of the T and DD
series. A method was developed for concentrating and purifying the phages.


Babov, D. M., Gubenko, L. T., Muromtseva, A. A., Yarotskaya, N. E. (1971).
Enterovirus Circulation in the Environment and Among the Population of
Odessa. HYG & SAN IT, 36(3): 388-91.

     Summary appeared  in 1971 edition of these abstracts.. The paper was
published originally in GIG SANIT. 36:54-7(1971).

Bagdasaryan,  G.  A.,  Abieva,  R. M. (1971). Survival of Enteroviruses and
Adenovirusesin Water. HYG & SANIT, 36(3):333-8.

     Summary appeared  in 1971 edition of these abstracts. The paper was
published originally in GIG SAN IT, 36: 10-14 (1971).

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Bell,  J.  A. (1972).  Viruses and  Water Quality.  J  AMER MED ASSOC,
219(121:1628.

      The hazard presented by viruses in sewage and water, the effectiveness of
treatment processes in  removing them, and  the  development of detection
methodology are discussed.


Bricout,  F. (1972). Viral Infections: Dissemination and Epidemiology.  NOUV
PRESSEMED,!: 1643-7. French.

      The roles of  water, food, and contact are discussed in a general review of
viral epidemiology.


CaJabro,  J. F., Cosenza,  B. J.. Kolega, J. J. (1972). Bacteriophages Recovered
From Septage.  J WATER POLLUT CONTRL FED, 44(12):2355-8.

      Two morphological phage  forms were recovered  from septage, one a
short-tailed variety that infected Gtrobacter  freundii, Escherichia coli, and
cell-wall  mutants  of Salmonella typhimurium, and the other a long-tailed
variety specific for Shigella flexneri. Only rough host strains were susceptible.
      Concentrations of linear alkyl sulfonate normally occurring in septage
did not effect the phage noticeably.


Cerkinskij, S. ISL, Trahtman,  ISI. (1972). The Present Status of Research on the
Disinfection of Drinking Water in  the USSR. BULL WHO, 46(2): 277-83.

      Recent  research  in  the  USSR aimed  at  evaluating  methods  for
disinfecting drinking water and at  determining the mechanisms of microbial
inactivation are reviewed. The advantages and disadvantages of chlorine, ozone,
and gamma irradiation are discussed, as are their effects on Enterobacteriaceae
and on enteroviruses.
Cherkinsky, S. N., Lovtsevich, E. L.. Ryabchenko, V. A. (1971). The Sanitary
Significance  of  Escherichia coli  in  Various  Conditions  of Water
Decontamination  in Respect to Enteroviruses. HYG & SAN IT, 36(3): 329-33.

     Summary appeared in 1971 edition of these abstracts. The  paper was
published originally in GIG SANIT, 36: 7-10(1971).

diver, D. O., Herrmann, J. E. (1912). Proteoly tic and Microbial Inactivation of
Enteroviruses. WATER RES, 6(6):797-805.

     Some  enteroviruses are sensitive to proteolytic enzymes. Coxsackievirus
A9  was  inactivated  by  proteolytic  bacteria,  notably  by Pseudomonas
aeruginosa.  This  inactivation  could  be  distinguished  from  adsorption or

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aggregation of the virus particles. 14C label from the viral coat protein, but not
32P from the viral nucleic acid, was taken up by the bacterial cells.


Dardanoni,  L.  (1971). Grculation of Wild Polioviruses.  ANN  SCLAVO,
13(5-6): 353-61. Italian.

     Circulation of wild polioviruses in Italy was greatly reduced by the mass
vaccination programs with attenuated poliovaccines which began in 1964.
     Besides  the  dramatic  reduction  in  reported  cases  of  paralytic
poliomyelitis, a  great reduction  in poliovirus recovery from man and sewage
occurred.
DiGirolamo, R.. Liston, J., Matches, J. (1972). Effects of Irradiation on the
Survival of Virus in West Coast Oysters.  APPL MICROBIOL, 24(6): 1005-6.

     Poliovirus  1 survived 400 krads of gamma radiation from a cobalt source
in whole and shucked oysters. From 7 to 13% of the virus survived this dose.
     Taste  tests with  shellfish that had undergone 400 krads of irradiation
revealed  that  the  oysters had undergone organoleptic changes  that  had
rendered them unpalatable.


DiGirolamo, R., Wiczynskj,  L., Daley, M.r Miranda, F. (1972). Preliminary
Observations on the Uptake  of Poliovirus by  West Coast Shore Crabs. APPL
MICROBIOL, 23(1): 170-1.

     West Coast shore crabs  (Pachygrapsus sp. and Hemigrapsus sp.), when in
seawater  contaminated  with  poliovirus  1 or  when allowed  to  feed on
virus-contaminated mussels (Mytilus califomianus), accumulated large amounts
of the virus.
DiGirolamo,  R., Wiczynski, L., Daley, M., Miranda, F., Viehweger, C. (1972).
Uptake of Bacteriophage and Their Subsequent Survival in Edible West Coast
Crabs After Processing. APPL MICROBIOL, 23(6): 1073-6.

      Edible  West Coast crabs fCancer magister and Cancer antennarius), when
in seawater contaminated with coliphage T4, accumulated large amounts of the
virus.
      From 2.5 to 20% of coliphage T4 survived boiling for varying periods in
contaminated crabs.
      Heat penetration studies showed that, although internal temperature in
the crabs was sufficient to inactivate the virus, the processing times normally
used to cook crabs were  not. These results suggest that processed crabs may
serve as vectors for the dissemination of virus diseases if the crabs are harvested
from a polluted area.

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Dragan. L. P., Goregliad, N. I. (1972). Distribution of Adenoviruses Among the
Population of Kiev in 1967-70, Based on  Studies of Sewage. MIKROBIAL
ZH, 34(1-2J:49-50. Translation presently not available. Ukrainian.

Ellender,  R. D., Morton, F.r Whelan, J.r Sweet, B. H. (1972). Concentration of
Virus from Water by Electro-Osmosis and Forced-Flow Electrophoresis. II.
Improvement of Methodology and Application to Tap  Water. PREP BIOCHEM,
2(3): 215-28.

      A modified electroosmosis (EO) method was devised for concentrating
viruses from  distilled and tap  waters. Selective membranes, which allow high
rate water transport, demonstrated the importance of membrane technology to
the  EO   procedure. The other modifications  added were  only secondary
contributory  factors.
      With the technic as developed thus far, poliovirus 1, seeded into distilled
or tap water at levels as low as 0.01 PFU/ml, was recovered.  Reovirus, seeded at
a level of 0.1 PFU/ml, was recovered. Physical concentration of diluent in all
experiments  averaged  40-fold.  Problems   were  encountered when
virus-containing  tap water was concentrated, most  likely, because of the high
content of dissolved impurities  which  may increase  viral  inactivation or
aggregation.
      EO  was more efficient than  forced-flow electrophoresis as a dehydration
procedure. However, the turbulent anode back-fractionation method did allow
for an increase  in the concentration  of virus. Lower virus inputs may have
accounted for  the decrease in virus recovery compared with that in  our
previous studies where higher inputs were used.

Ellender,  R.  D.,  Sweet,  B.  H.  (1972).  Newer  Membrane Concentration
Processes and Their Application to the Detection of Viral Pollution of Waters.
WATER RES, 6(6):741-6.

     New classes  of ultrafilter membranes (both cellulosic and synthetic
polymeric varieties) for concentrating viruses from distilled water were  tested
in a model rapid flow, recirculating, thin channelized  system under a positive
pressure   gradient (osmotic or  compressed  gas).  In  contrast to  the
adsorption-elution  systems (membranes and  polyelectrolytes),  the method
depends upon rapid transport  of water and low molecular weight substances
through the  membrane and rejection of viruses and other high molecular
weight materials. Because of flow rates, turbulence  factors, and  membrane
properties, virus adsorption is minimized.
     Polymeric  membranes offer greater  fluxes  (dehydration rates) than
cellulosic  membranes. In  tests  with up to 20 liters of distilled water, greater
than 50%  recoveries of poliovirus 1 were obtained when virus inputs were 0.01
to 1 PFU/ml. Dehydration factors were 400X or greater. The reasons for virus
loss are not understood, but may involve adsorption factors associated with the
mechanical portions of the system.

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England, B. (1972). Concentration of Reovirus and Adenovirus from Sewage
and Effluents by Protamine Sulfate (Salmine) Treatment. APPL MICROBIOL,
24(31:510-12.

     Protamine sulfate precipitated  adenoviruses and reoviruses from sewage
and  effluents.  Bovine  albumin increased the efficiency of recovery of these
viruses. The viruses were efficiently eluted from the precipitate with IN NaOH.
Eighty to 100% of exogenously added virus was recovered with this technic.
Most enteroviruses were recovered inefficiently or not at all by this method.
     The protamine precipitation  method was at best as  effective  as the
Al(OH)3 technic for recovering naturally occurring reoviruses from sewage and
effluents, but superior to  the insoluble polyelectrolyte (PE60)  and CaHPO4
adsorption methods.
     Adenoviruses and reoviruses, frequently overgrown in plaque assays, were
more readily recovered  by tube assay methods.

 Ergasheva,  L.  E., Artykov, M.  S., llyinsky, I.  I. (1972). Sanitary-Virologic
Features of Sewage in Certain Urban Sewerage Systems of Uzbek SSR. GIG
 SANIT, 37(9):90-1. Translation presently not available. Russian.
 Farley. C. A., Banfield, W. G., Kasnic, G., Jr., Foster, W. S. (1972). Oyster
 Herpes-Type Virus. SCIENCE, 178(11):759-60.

      A herpes-type virus infection, the first  to be  found in  an invertebrate
 animal, is reported in the oyster Crassostrea virginica. Intranuclear herpes-type
 viral inclusions  were  more  prevalent in  the  oyster  at   elevated water
 temperatures of 28 to 30 C than at normal ambient temperatures of 18 to 20
 C.  The  inclusions were  associated  with  a  lethal disease  at the elevated
 temperatures.


 Foliguet,  J.-M.,  Doncoeur, F.  (1972).  Inactivation  of Enteroviruses and
 Salmonella in  Fresh  and Digested  Wastewater  Sludges by  Pasteurization.
 WATER RES. 6(11): 1399-1407.

      In laboratory studies with seeded viruses and bacteria, thermal treatment
 of fresh and digested sewage sludges resulted in pasteurization. Reductions of
 more than 99.99% of  poliovirus  1 and more than 99% of coxsackievirus B3
 were obtained at 80 C in  10 minutes.  The heat-up time was less  than  10
 minutes. Even greater reductions of Salmonella paratyphi B were demonstrated
 under these test conditions.
 Francis, T. I., Wright, S. G., Onukogu, A. I., Okafor, E. E. (1972). Clinical and
 Epidemiological Studies of an Epidemic of Jaundice in Aba and Onitsha, 1970.
 W AFRICAN MED J, 21(4):43-6.

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      Eighty-four cases of jaundice occurred in two towns in the East Central
State of Nigeria. Most of the cases were clinically compatible with infectious
hepatitis, but a small group of patients had symptoms suggestive of mild yellow
fever or other group B arbovirus infections, and a smaller group had hepatoma.
These clinical findings were confirmed by serological data.
      Epidemiological data suggested that the infectious hepatitis outbreak was
waterborne.
Furuse, K., Watanabe, I. (1971). Effects of Ultraviolet Light (UV) Irradiation
onRNA fhageinH2Oandin D*O. VIROLOGY, 46(10): 171-2.

     The UV sensitivities of MS2 and Q/3 phages were slightly  different in
H20 than in D2O. There was little difference between the UV sensitivities of
the whole phages and those of their RNA freed from coat proteins. Thus, the
coat proteins apparently did not protect the  nucleic acids of these phages
against absorption of UV energy.
     The inactivation of RNA phages by UV irradiation may be the result of
pyrimidine hydration and  pyrimidine dimerization or  to changes in RNA
configurations resulting from the secondary effects induced by hydration or
dimerization.
Garibaldi, R. A., Murphy. G. D. Ill, Wood, B. T. (1972). Infectious Hepatitis
Outbreak Associated with Cafe Water.  HSMHA HEALTH REP, 87(2): 164-71.

     In May, June,  and July 1970, 95 cases of infectious hepatitis occurred
among residents of Polk County, Arkansas, and in surrounding counties in the
State and in Oklahoma. Of the 95 patients, about 80% had patronized a cafe in
Hatfield, Arkansas between mid-April and the end of July 1970. In a group of
470 area residents who  had not been ill, only 19% had patronized the same
cafe.

     Epidemiologic  and laboratory evidence incriminated cafe water  as the
most  likely  source  of  the epidemic.  Of  78 hepatitis patients  who  had
patronized the cafe, 97% had drunk water as compared with 55% of 53  non-ill
patrons questioned. Although sanitary conditions inside the cafe appeared to be
excellent, the tap water  was contaminated with coliform bacteria. Fluorescein
dye flushed down the toilet in the cafe appeared in the tap water 20 days later.
Lateral  seepage  of effluent from a septic  tank through underground shale
fissures  appeared to be  the most likely  route in contamination of the cafe
water.
     The  cafe  was closed voluntarily  by the proprietress in  June,  and
household  contacts  were  treated  with immune  serum globulin. An
immunization campaign  for area residents was carried out in July to reduce the
extent and  severity of secondary spread.

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Gentry, R. F., Braune, M. O. (1972). Prevention of Virus Inactivation During
Drinking Water Vaccination of Poultry. POULTRY SCI, 51 (4): 1450-56.

      Newcastle disease (ND) and infectious bronchitis (IB) drinking water
type vaccines secured at the time of field application had sufficient viable virus
to produce an  immune response. IB vaccine with only 0.7 EID at the final
drinking water concentration stimulated an immune response in  4-week old
chickens.
      IB virus  liters were  reduced  in filtered tap  water, both at  room
temperature and in an ice  water bath. ND virus liters were also markedly
reduced  in  filtered tap  water at room  temperature.  Powdered  skim milk
protected against inactivation.
      One mg/liter  of chlorine or quaternary ammonium sanitizer inactivated
ND, IB, and avian encephalomyelitis viruses. Skim milk neutralized the chlorine
and the quaternary ammonium salt.
      Virus  neutralization  tests  on serums from 34 field flocks  vaccinated
against ND and IB without skim milk in the drinking water showed that 32%
had  not responded to the  ND vaccine  and 44% were  still susceptible  to IB.
When skim milk had been  added to the drinking water, all birds in 42  flocks
had an immune response to the ND vaccine, and only 2 (4.8%) failed to respond
to IB vaccine.
      The results indicated the value  of adding skim milk to drinking water
used for vaccinating chickens.

Globa,  L. I., Lastovets, L. M., Rotmistrov, M. N., Golub, N. F., Radolits'ka, L.
S. (1971). Adsorbing Materials for Virus Removal from Water. DOPOV AKAD
NAUK  UKR SSR. Ser. B, 33:1036-8. Ukrainian.

      Polygoskite, bentonite, aglaporile, vermiculite, permutile, pyrophillite,
gypsum, and silica gel adsorb viruses from water, and may be used for that
purpose prior to coagulation in water purification.

Globa,  L. I., Lastovets, L. M., Rotmistrov, M. M. (1972). Ability of Minerals to
Adsorb  Viruses from  Water.  MIKROBIOL  ZH.  34(1-2):64-5.  Translation
 presently not available. Ukrainian.


 Gloyna, E. F. Disease Transmission Control as a Factor in Pond Design. In
 "Waste Stabilization  Ponds," Chapter 5, WHO MONOGRAPH SER, 60 (1971),
 93-108.

      Treatment facilities are not yet designed specifically for removing viruses
and  other  pathogens, but more engineers and scientists  are beginning to
recognize that there is a problem.
      The removal  of BOD without regard to the destruction of disease-causing

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 agents is not enough. Pathogens must be destroyed in the wastewater treatment
 plant or diluted to a negligible risk level in the environment.


 Grabow,  W.  O.  K.,  Nupen,  E.  M.  (1972). The  Load  of Infectious
Micro-Organisms in The Waste Water of Two South African Hospitals. WATER
 RES, 6(12): 1557-63.

     Counts of microorganisms including pathogenic bacteria, viruses, and
 parasite  ova in the sewage of two hospitals were compared with those in city
 sewage.
     One  of these hospitals is an isolation facility with stringent disinfection
procedures. Counts of  all  the   organisms  tested,  except  Pseudomonas
aeruginosa,  were lower in the sewage  of this hospital than in the sewage
effluent  of the town in which it is located. Hie other hospital is a general
hospital  with less stringent disinfection procedures. Counts of some organisms
were slightly higher in the sewage of this hospital than in the sewage effluent of
 the city in which it is located.
     Under normal conditions  the wastewater of hospitals  may  need no
treatment before  disposal into city sewers. The discharge of primary hospital
effluents into streams or their use for irrigation, however, should be considered
with care.


Gromashevsky, L V. (1972). Evolution of Viruses and Viral Diseases.  ZH
MIKROBIOL  EPIDEM  IMMUN,  12(12):48. Translation presently  not
available. Ukrainian.


Grushko, Y. M.  (1971). Non-Soviet Journals on  Water Pollution. HYG &
SANIT, 36(2):251-3. (Same ^Problems ofSanitaryProtection of Water Bodies
in the Pages of Foreign Journals, GIG SANIT, 36:73-5. Russian.)

     This  paper is a  brief review of water pollution control literature,
including some dealing with the  viral problem, published in non-Russian
journals.
Hall, M W., Sprout, O. J. (1971). Water Quality and Recreational Land Use.
PUBLIC WORKS, 102fc):S2-6.

     The increased demand for water-based recreation creates problems for
water  pollution  control authorities.  Degradation  of surface  waters  by
water-borne pollutants originating in shore-side camps and homes has received
little attention. Disposal of treated wastewater which contains viruses  and
soluble  plant nutrients that may be transported farther through the soil than
organic materials and bacteria creates pollution problems for lakes and streams.
     These problems are reviewed in some detail.

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Hill, W. F., Jr., Akin, E. W., Benton, W. H., Metcalf, T. G. (1972). Virus in
Water. II.  Evaluation of Membrane Cartridge Filters for Recovering Low
Multiplicities of Potiovirus from Water. APPL MICROBIOL, 23(5):880-8.

     Poliovims 1, added in large numbers to  100 gallons of  tap or estuarine
water containing 1,200 Mg Mg^/ml, were recovered completely in aMillitube
MF cartridge membrane filter. Four elutions of 250 ml each with 5 X nutrient
broth in 0.05  M carbonate-bicarbonate buffer at pH 9.0 recovered the virus
from the cartridge. When 20 to 40 PFU of virus were added to 100 gallons of
tap or estuarine waters, a two-step concentration procedure was used for virus
recovery. Concentration first by the Millitube MF cartridge procedure followed
by secondary concentration of the eluate by the two-phase polymer procedure
resulted in virus recoveries of up to 67%.


Jordan, F.  T. W., Massar, T. J. (1971). The Influence of Copper on the Survival
of Infectious Bronchitis Vaccine  Virus in Water. VET REC, 89<12):609-10.

      Infectious bronchitis virus  (chickens)  survived longer in waters that did
not have prolonged contact with copper pipes than in waters that did.
      In laboratory experiments, the virus survived for more than six hours in
water that  contained less than 0.02 mg/liter of Cu++ (as CuSCU), but only for
two hours  in similar water containing 0.2 mg/liter of Cu"1"1".  The addition of
iron filings (to displace the Cu"1"*") to the water, or the addition of powdered
skim milk  to a concentration of 0.1% increased virus survival time to at least
six hours.
 Kiseleva,  L.  F.  (1971).  Survival of Polioviruses,  Echoviruses,  and
 Coxsactteviruses in Food Products. VOPR PITAN, 30(11-12):58-61. Russian.
      Enteroviruses survived for a long time in large numbers in tap water, milk,
sour  mflk  products,  and bread. Hence,  food products contaminated with
enteroviruses may facilitate their spread among the population.
 Konowalchuk,  J.,  Speirs,  J.  I.  (1972). Enterovirus Recovery from
Laboratory-Contaminated Samples  of Shellfish.  CAN  J  MICROBIOL,
 18(7): 1023-9.

      A sensitive  reproducible  method for recovering enteroviruses from the
 supernatant fluids of centrifuged homogenates of laboratory-contaminated
 shellfish meats is  described. The supernatant fluids contained varying amounts
 of toxic materials that interfered with coxsackievirus B5 recovery in HEP-2
 cells.
      Most of the cytotoxicity was removed by precipitation with hydrochloric
 acid at pH 3.0 to 3.5. The acid-treated samples were diluted 1:4 in fetal bovine

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serum and then mixed with cell suspensions to adsorb the viruses which were
subsequently enumerated as plaques.
      Hie method is applicable to oysters and especially to mussels and clams
because  of the high  cytotoxicity of  these shellfish.  Other methods  for
concentrating  sample  fluids were also  examined as a means for detecting
enteroviruses in naturally contaminated shellfish.

Korsh,  L.  E.,  Bagdasaryan, G.   A.,  Vlodavets,  V. V.  (1972).
Sanitary-Microbiological Studies  in the Prevention of  Bacterial and Viral
Infection. VESTN AKAD MED NAUK SSR, 27{4):71-8. Translation presently
not available. Russian.
Kostenbader, K. D., Jr., diver, D. O. (1972). Polyelectrolyte FJocculation as
an  Aid to Recovery  of Enteroviruses from Oysters.  APPL MICROBIOL,
24(4): 540-3.

      A simple, rapid method for recovering enteroviruses from  oysters is
described. A polycation sewage flocculant promoted cohesion of oyster solids
and thereby aided  separation of these from the viruses. The suspension or
extract obtained was inoculated directly into cell cultures or concentrated first
for greater sensitivity.
      Recovery of 80 to 100% of experimentally inoculated virus was achieved.

Lautier, F., Lavillaureix, J. (1972). Recovery of Viruses From Polluted Waters,
Gty Sewage, and from Urban and Rural Sewage  Treatment Plants.  PATH
MICROBIOL, 38(1):64-5. French.

      Sixty-four samples of sewage, seven of river water (111 River), and nine of
treated effluents, all from the city of Strasbourg, were tested for viruses by the
A1(OH)3 adsorption procedure. Enteroviruses were recovered from eleven of
the samples.


Lautier. F.. Wilenskj, A., Lavillaureix, J. (1971).  A Method forDetecting Viral
Particles in Polluted Waters. PATH MICROBIOL, 37:99-104. French.

      Viruses were  recovered from 100 ml volumes  of sewage effluent by
adsorption onto A1(OH)3.


Led ere, H.  (1971).   Marine and  Freshwater Pollution.  REV  INTERN
D'OCEANOGR MED, 24:155-70. French.

     Pollution  of  marine  and  fresh water environments by microbes is
discussed, with emphasis on  Vibrio, Qostridium, Salmonella,  Mycobacteria
and viruses. The risk of infection to humans and methods for the prevention of
microbial pollution are considered.

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Legier, F. (1972). Hygienic Problems in Swimmingand Camping Areas. OEFF
GESUNDHEITSWES, 34(6):317-37. German.

     Within  a discussion of the sanitary  requirements for swimming and
camping facilities, the transmission of virus diseases, including the transmission
of molluscum contagiosum and plantar warts, are noted.
Liebsdier, S. (1970). Enterovimses in Swimming Pool  Water. Z GESAMTE
HYG, 16(3): 198-200. German.

     Coxsackievirus B3 was recovered on  HEP-2 cells from 20% of samples
taken from  an outdoor swimming pool.  The virus caused  meningitis and
encephalitis in children. Bacteriological examinations of pool  water disclosed
high concentrations of coliforms.
     Application of 0.3 to 0.5 mg chlorine/liter is not adequate for swimming
pool disinfection.
Lindeman, S., Kott, Y. (1972). The Effect of Otlorination on Enteroviruses in
the Effluents of the Haifa Sewage  Treatment Plant. ISRAEL J MED  SCI,
     Two hundred and fifty ml of Haifa sewage effluent was passed through
cellulosic membrane  filters (0.45  j*m) and the filtrate, in turn, through an
alginate ultrafilter (10 nm). The ultrafilter was dissolved in 3.8% sodium citrate,
and viruses concentrated by this method were recovered by plaquing in primary
Cercopithecus  aethiops  kidney  cell  cultures.  About  35  to  45  PFU of
viruses/ 100 ml of effluent was detected  with this  method as compared with
only 3 to 5 PFU/ 100 ml with alum flocculation.
     When 8  mg/liter  chlorine were  applied to effluents for one hour, no
decrease occurred in virus numbers. However, after  two hours of contact, virus
numbers were decreased.
Long, W.  N.,  Bell,  F.  A.. Jr. (1972). Health Factors and Reused Waters. J
AMER WATER WORKS ASSN, 64(4):220-5.

     In a discussion of the possible health hazards associated with the reuse of
renovated sewage, the virus problem is analyzed in some detail.
Lund,  E. Inactivation  of Viruses^ In Proceedings of the 6th International
Conference on Water Pollution Research, June 1972. Pergamon Press Ltd.

     A number of chemical and  physical treatments may inactivate viruses.
Viruses may also become harmless spontaneously if kept outside proper cells
for a sufficiently long period.

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      For most inactivation processes, we  know little about  the  reactions
taking place. The virus genome may not be able to take command in a cell. The
virion may gain entrance into a cell, but fail to be stripped, or it may not get
into a cell at all, or, the infectious nucleic acid may truly become inactivated.
      Spontaneous inactivation of the small RNA viruses is a slow process in
water at a temperature of 20 C or below. It is not an oxidation process. If it is
the same process as thermal inactivation, it works faster on the proteins than
on the nucleic acids. Heat-inactivated virus may contain infectious nucleic acid.
      Viruses are inactivated faster at  basic pH values. Alkaline inactivation
may be the result of a process similar to thermal inactivation and thereby be
one of protein denaturation, the result of splitting and uncoiling of proteins.
      Spontaneous inactivation  is  accelerated in  sea  water. The  virus
inactivating capacity of sea water seems bound  to sea water, and is not found
in fresh water. Most workers point to one or more species of the marine
microflora.as the source of this inactivation. The nature of this process remains
essentially obscure.
      Viruses are sometimes more easily detected in primary sludge than in the
water.
      Chemical precipitation of sewage with calcium and aluminum  salts may
remove viruses  from naturally contaminated  waters, but the virus is essentially
not  inactivated.  Contrary  to what  might  be  expected,  virus  may  be
demonstrable even when the precipitate has been obtained at pH 10.5 to  11.0.
This must be considered in the disposal of sludge.
      In what  seems reasonable agreement  both with the laws of physical
chemistry and results empirically determined, the rate of inactivation  of viruses
by oxidizing disinfectants is dependent on oxidation potential.
Mack, W. N., Lu, Y. S., Coohon, D. B. (1972). Isolation of Poliomyelitis Virus
from a Contaminated Wett. HEALTH SERV REP, 87{3):271-4.

      The water  supply of a large restaurant was implicated as a source of
infection in the illness of several patrons. A search for bacteria in the food and
water source of the restaurant did not uncover the responsible agent.
      However, poliovirus 2 was recovered from five gallons of the restaurant's
well water concentrated by flocculation and ultracentrifugation.
Malherbe, H. (1971). Viruses in Water. S AFRICAN J SCI, 67(3)124-7.

      The problem of viruses in sewage and water, particularly as it applies to
the Union of South Africa, and the effectiveness of treatment technics  are
reviewed.

                                   12

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Markov, A. P. (1972). Experience in the Sanitary-Virological Examination of
Waste Waters. GIG SANIT, 37(2):94-5. Translation presently not available.
Russian.
Marks, H. C. Residual Chlorine Analysis in Water and Waste Water. In "Water
and  Water  Pollution  Handbook,"  Chapter  22,  edited by  L.  Ciaccio.
Marcel-Dekker, New York, New York (1972), 1213-47.

      The measurement of chlorine and its compounds in water and sewage are
reviewed in an extensive and detailed treatment of the subject.
Martin, A. E. (1972). Medical Considerations in the Abstraction of Potable
Waters from Polluted Sources. WATER TREAT & EXAM, 21(3):202-12.

     In a general review of the problem of pollution of water supplies, the
need to protect against virus contamination of supplies, the means for doing so,
and the standards designed for that purpose are discussed.
Morris, J. C. (1971). Odorination and Disinfection-State of the Art. J AMER
WATER WORKS ASSN, 63(12): 769-74.

     The state of  the  art of chlorination and disinfection with ozone,
bromine,  iodine,  and  other halogen  species  is  assessed.   The virucidal
effectiveness of chlorine and some of its compounds is discussed. The relative
value of ozone and the other halogens in different need situations is explored.
Morse, L J., Bryan, J. A., Hurley, J. P., Murphy, J. F., O'Brien, T. P., Wacker,
W. E. C. (1972). The Holy Cross College Football Team Hepatitis Outbreak. J
AMER MED ASSOC, 219(6):706-8.

     During a 15-day period in September and October 1969, an outbreak of
infectious hepatitis affected the members of a college  football  team. Of 97
persons exposed,  90 were  infected. Thirty-two  experienced typical icteric
disease, 22 were anicteric but symptomatic, and 36 asymptomatic players had
significantly elevated serum glutamic pyruvic transaminase values (>100
units). Other athletes, using the same facilities but arriving six days after the
established date of exposure, were unaffected.
      The decision to obtain blood samples from the entire team as soon as the
initial cases were recognized resulted in the demonstration of an unexpectedly
high attack rate of 93%. Epidemiologjc investigation revealed that an infected
group of children  in the neighborhood, an imperfect drinking water supply, a
warm August day, a football team in training, and a local fire were links in the
chain which resulted in this most unusual outbreak of infectious hepatitis.

                                   13

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 Mosley, J. W. (1972). Viral Hepatitis:  A Group of Epidemiologic Entities.
 CAN MED ASSOC J , 106,{2)427-34.

      This paper comprises a thorough sophisticated analysis and discussion of
 viral hepatitis including waterborne infectious hepatitis.


 Nestor, I., Gostin, L (1971). Dissemination of Enteric Viruses in Urban Areas.
 STUD CERCET INFRAMICROBIOL, 22(3):209-14. Rumanian.

      Surveys carried out during the 1962-1969 period on the presence of
 enteroviruses hi  sewage, rivers, and in water supplies in towns and in other
 environments are summarized. Viruses were concentrated with ion exchange
 resins, and  with  the Saccharomyces cerevisiae, and aluminum hydroxide
 adsorption methods.


 Nikolaevskaya,  Z.  S., Aizen,  M. S.  (1972).  Detection of  Minimal
 Concentrations  of Viruses  in Large Volumes By  UltrafUtration  Through
 Soluble Lanthanum-Aktminum-Alginate   Ultrafttters.  VOP  V1RUSOLOGII,
 6:723. Russian.

      Double filtration through bacterial  asbestos  filters treated with 0.05%
 aqueous sodium  alginate followed by  concentration by ultrafiltration through
 soluble La-Al-alginate  ultrafilters facilitated recovery of small  amounts of
 viruses from large volumes of water. The concentration factor was 400 to 500.
 There appeared to be no loss of viruses.
      With this  method,  enteroviruses and other  cytopatnic agents  were
 recovered  from  37%  of 1-liter effluent samples taken  from a plant on the
 Moskva River, and from 37% of 3-liter river water samples taken at Kalinin on
 the Volga  River


 Okun,  D. A.  (1972). Safe  Drinking  Water.  AM  J  PUBLIC HEALTH,
 62(7):903-4.

      Community drinking water supplies should be developed from the best
 sources available to minimize the risk  of transmission of viral and other
microbial diseases and to minimize the ingestion of toxic and other hazardous
substances.
Olifson, L. E., Baltenko, E. N., Bukharin, O. V., Pozhar, V. N., Turovets, G. L.
(1970). On the Feasibility of Comprehensive  Utilization of Industrial  and
Domestic  and  Fecal Effluents  in a Mining  Combine.  HYG  & SAN IT,
35(12}:412-13.  (Same  as  Utilization of Industrial and Agricultural
Wastewaters, GIG SAN IT, 35(12):92-4. Russian.)

      Poliovirus 2, echovirus 7, and coxsackievirus B3 survived for three to 15

                                  14

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days in acid (pH 2.0) and alkalinepH (11.0-13.5) industrialwastewaters from
mining and ore processing operations.

Palfi, A. B. (1971). Virus Content of Sewage in Different Seasons in Hungary.
ACTA MICROBIOL ACAD SCI HUNG, 18(4):23l-7.

     Three hundred  and seventeen viruses were recovered from 336 sewage
samples collected in Hungary in 1969. Sixty-two were polioviruses of vaccine
origin.  Of the remaining 255 strains, 43% were reovirus 1,21% were echovirus
7, and  18% were  echovirus  11.  These  three virus  types  were prevalent
throughout  the  year.  The  other  strains  recovered  were identified  as
coxsackieviruses Bl, B3, and B4, and echoviruses 1, 6,12,14,19, and 20.
     The greatest number  of viruses were  recovered  in  August.  Fewer
recoveries were made during October, November, and September. The number
of recoveries  was lowest  in  February and  March, following compulsory
vaccination programs.
     In 1968, reovirus 1, coxsackieviruses Bl and B3, and echoviruses 6 and 7
predominated.

Pana, A. (1971). Concentration of Enteroviruses from Water by Adsorption on
Insoluble Potyelectrolytes. NUOVI  ANN IG MICROBIOL, 22(11-12):4l5-27.
Italian.

     The insoluble  polyelectrolyte method yielded better  virus recoveries
from surface water than the A1(OH)3 adsorption or alginate filter methods.

Pavoni, J. L, Tittiebaum, M. E., Spencer, H. T.f  Fleischman, M., Nebel, C.,
Gottschling, R. (1972). Virus Removal from Wastewater Using  Ozone. WATER
& SEWAGE WORKS, 119(12):59-67.

     Seeded coliphage f2 was  stable in secondary effluent for at least five
hours, and was unaffected by flow or mixing in an ozone reactor.
     The virus was inactivated  completely in  five minutes by a total ozone
dosage of approximately 15 mg/liter  which left a residual of 0.015 mg/liter.
The  rate of  inactivation  was greater  for   f2  phage than  for  bacteria.
Carbonaceous material  reacted with ozone more slowly than  the phage, also.
The  mechanism of destruction of bacteria and viruses was probably oxidative.
Ozone  may  be a general cytoplasmic oxidant which causes cell  lysis and the
release of soluble COD.
     Standards for  ozone use in wastewater are needed. Current disinfection
standards are concerned only with chlorine. The contact times and dosage
levels for chlorine are not applicable to ozone. Escherichiacoli, presently the
only indicator of disinfection efficiency, cannot be used always as a virus
indicator.

                                  15

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Poduska,  R.  A.,  Hershey,  D. (1972). Model for  Virus  Inactivation  by
Ottorination. J WATER POLLUT CONTR FED, 44(5):738-45.

      A mathematical model to explain the process of virus inactivation by
chlorination has been developed and is based on the principles of first-order,
irreversible chemical kinetics for the  reaction between chlorine and virus
particles in solution. The viruses studied were poliovirus 1 and coliphages f2
and MS2.
      Inactivation is dependent on the time of contact between viruses and
chlorine, the concentration of chlorine, and the pH of the system. The process
is complicated by the clumping of viruses which results in  a heterogeneous
inactivation system. Each of these factors was included in the model in order
to quantify the kinetics of inactivation.
      In the  mathematical analysis, the virus clump sizes were numbered from
1 to N, beginning with clump size 1.  The  model consists of N independent
exponential terms that describe the inactivation of each of the clump sizes in a
system. An  inactivation  rate  constant  is  associated with  each term,  and
characterizes the rate of inactivation of each clump size.


Powers, E.  L, Gampel-Jobbagy, Z.   (1972). Water-derived Radicals  and
Radiation  Sensitivity of Bacteriophage 77. INTJ RADIATBIOL, 21(4):353-9.

      Ethanol  scavenging  of -OH and *H in buffered, X-irradiated suspensions
of coh'phage T7 saturated with either N2, N2 O, or O2 resulted in partial sparing
of the virus. In pure suspension, therefore, some of the inactivation of T7 phage
appears to drive from-OH activity.
      Radiation-induced sublethal  damage  to the phage  may play a part in
determining sensitivity.
Rao, V. C., Chandorkar, U., Rao, N. U., Kumaran, P., Lakhe, S. B. (1972). A
Simple Method for Concentrating and  Detecting  Viruses  in  Wastewater.
WATER RES, 6( 12): 1565-76.

      A modified membrane filter method was developed for recovering viruses
from sewage and sewage effluents. Small amounts of enteroviruses seeded into
autoclaved sewage were adsorbed onto 0.45 pm, 47 mm diameter membrane
filters  at  pH  3 and eluted  at  pH 8.  Clarifying  the sewage  first by
homogenization for four minutes in a Waring blender and centrifugation  at
1,800 g and 9,230 g facilitated filtration with no loss of virus. Complete
recovery of the seeded virus was achieved.
     Enteroviruses  seeded into fecal suspensions (BOD = 600 mg/liter)
adjusted  to pH 3  and containing 1,200 mg Mg++  (as MgCl2)/liter  were
completely recovered.
     During a  one-year program  of monitoring raw sewage from a middle

                                  16

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 income community in Nagpur, a maximum of 3,150 PFU of vimses/liter were
 recovered during the monsoon season and 11,575 PFU/liter were recovered
 during the winter.


 Rowland, A. J., Skone, J. F. (1972). Epidemiology of Infectious Hepatitis BR
 MED BULL, 28(2): 149-55.

      The epidemiology, distribution, prophylaxis, and therapy of infectious
 hepatitis are discussed.
 Ruschi, A. (1971). Presence and Epidemiological Significance of Polioviruses in
 Sewage in Ksa (Italy). ANN SCLAVO, 13(5-6): 370-6. Italian.
      The frequency of poliovirus recovery from the sewage of Pisa in 1962
 and 1963, prior to mass live-poliovirus vaccination programs, peaked during the
 summer months.
      In 1965 and 1966, subsequent to the initiation of vaccination programs,
poliovirus recovery from sewage peaked during the winter months.
      Other enteroviruses were recovered with about equal frequency during
 both sampling periods, with peak  recoveries occurring  during the  summer
 months.


 Scarpino,  P.  V.,  Berg, G.r  Chang,  S.  L, Dahling,  D., Lucas, M. (1972). A
 Comparative Study of the Inactivation of Viruses in Water by Chlorine. WATER
 RES, 6(8): 959-65.

      The inactivation rates  of poliovirus  1 and  Escherichia coli  by
hypochlorous acid (HOC1)  in phosphate buffer (pH 6), and by hypochlorite
 ion (OC1~) in borate buffer (pH 10) were determined at 5  C.
     Hypochlorite ion  inactivated poliovirus 1  about seven times more rapidly
than HOC1, whereas HOC1 inactivated E. coli about 50 times more rapidly
than OC1~~. Thus, poliovirus 1 was inactivated more rapidly at pH levels where
the free chlorine  was in the form of OC1~ rather  than  in the form  HOC1.
Alkalinity at pH 10 did not inactivate poliovirus 1 during the test period.
     A reassessment of the  chemistry of HOC1 ionization may be in order to
determine  whether  borate  buffer  alters the  equilibrium of the  reaction
suppressing ionization, or whether it brings about the formation of virucidal
forms heretofore undescribed.
Shah, P. C., McCamish, J. (1972). Relative Chlorine Resistance of Poliovirus 1
and Coliphages f2 and T2 in Water. APPL MICROBIOL, 24{4):658-9.

     A mixture of ammonia chloramines  inactivated  poliovirus 1  and
coliphage T2 more rapidly than they inactivated coliphage f2.

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Shane,  M. S., Cannon,  R.  E.,  DeMichele,  E.  (1972). Pollution Effects on
Phycovirus and Host Algae Ecology. J WATER  POLLUT CONTROL FED,
44(12): 2294-2302.

      Algal viruses of Plectonema boryanum were recovered from polluted
stretches  of the Christina River, but not from the unpolluted headwaters.
Recovery of the viruses correlated well with chemical, physical, and biological
pollution indicators.
Shelton, S. P., Drewry, W. A. (1971). Virus Removal by Chemical Coagulation.
Water  Resources Research Center No.  15, Department of Civil Engineering
Research Series No. 12, University of Tennessee, 121 pages.

     In waste  and river waters, aluminum sulfate, ferric chloride, and ferric
sulfate were effective coagulants, removing more than 99% of seeded coliphage
f2  in   at  least  one   of the  two waters. Cationic polyelectrolytes were
unsatisfactory as primary coagulants because they did not form good floes.
Cationic, nonionic, and  anionic polyelectrolytes improved coagulation with
aluminum  sulfate;  however, the value of the  polyelectrolytes is doubtful
because of their cost. Sodium aluminate, in conjunction with  aluminum
sulfate, was  effective for virus removal. Optimum dosages of the two  for
turbidity and COD reduction, however, did not correspond to optimum
dosages for virus removal.
     A simple mathematical relationship may exist between raw water COD
and the optimum dosage of primary coagulants for virus and turbidity removal.
     The colloidal titration technic and the Zeta-Meter methods for locating
isoelectric points did not always produce the same results, and the optimum
dosages for all parameters often did not correlate well with either method for
isoelectric point location.
Shitskova, A.  P.  (1971).  Hygienic Aspects of Preventing Digestive  TYact
Diseases.  GIG  SANIT,  36(12):7l-7. Translation  presently  not available.
Russian.
Shuval, H. I., Katzertelson, E. The Detection of Enteric Viruses in the Water
Environment.  In  "Water Pollution Microbiology," Chapter 15, edited by  R.
Michael, Wiley-lnterscience, New York, New York (1972), 347-61.

      Enteric virus diseases, especially infectious hepatitis, may be waterborne.
Outbreaks may be epidemic or sporadic. Methods for evaluating the virological
safety of water supplies need to be developed.
      Many enteroviruses survive longer than coliforms in rivers and in sea
water. These viruses are also more difficult to remove or inactivate by sewage
and water treatment methods than coliforms.

                                   18

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     Thus,  efficient  methods for detecting  viruses in water need to be
developed. For  this  purpose,  adsorption  to  particulates, adsorption onto
membrane filters, and phase separation are the most promising methods at this
time.
Singh, K, V., Bohl, E. H. (1972). The Pattern  of Enteroviral Infections in a
Herd of Swine. CAN J COMP MED, 36(7):243-8.

      Viruses were recovered in porcine kidney cell cultures from the feces of
26 pigs, 34 to 64 days old, in a healthy herd of swine. The pigs within each of
four litters infected began shedding viruses in their feces at about  the same
time.  The type of virus initially recovered was usually the same.
      Subsequently, waves of infection with different enteroviruses occurred
during the following  six months. At least six antigenically different viruses
were recovered from the herd over a 26-month period. Most, and perhaps all,
of these viruses apparently belonged to the enterovirus group. No disease was
associated with the infections.
      The colostrum  and milk of sows  contained significant  amounts of
enteroviral antibodies. Prior to nursing, the serums of new-born pigs contained
no  enteroviral  antibodies, but shortly  after  nursing  high tilers of  such
antibodies developed. Antibodies were detected also in  the feces of suckling
Pigs-
Slack, J. G. (1972). Water Reclamation from Sewage Effluent:  Experimental
Studies in Essex. WATER TREAT & EXAM, 21(3):239-58.

      Full-scale  plant tests  in  a standard  inverted pyramidal upflow tank,
at a particle-settling velocity of 0.5 mm/sec, showed that about 99 to 99.9%
of  the  viruses  in  sewage  and  about 95  to  99% of  the  Escherichia coli
and other coliforms present were removed by treatment with 150  mg/liter
of  aluminum sulfate and 250 mg/liter of powdered carbon.

Sorter,  C. A., Malina. J. F., Jr., Sagik, B. P. (1972). Quantitative Procedure for
Evaluating the Performance of Water and Waste Water Treatment Processes at
Naturally Occurring Virus Levels. ENVIRON SCI & TECHN, 6<5):438-41.

      A modified, standardized insoluble polyelectrolyte technic was used to
evaluate the removal of coliphage T2 from water and wastewater at levels down
to 1 x 10"4 PFU/ml. Extrapolation of the results below this level is possible if
sufficient quantities of water are available.
      The  efficiency  of  the  method  for  concentrating the  virus is
pH-dependent.  At  the optimal  pH, a constant virus recovery of 25% was
achieved at phage concentrations of 103 to 10~4 PFU/ml.

                                   19

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Sorter, a A., Malina, J. F., Jr., Sagik, B. P. (1972). Virus Rejection by the
Reverse Osmosis-Ultrafiltration Processes. WATER RES, 6(11): 1377-88.

      Asymmetrical cellulose  acetate membranes of  the type  used  in
ultraffltration  and reverse  osmosis rejected from  99.2%  to  over  99.999%
coliphage T2 and a poliovirus present in feed solutions of PBS or lake water.
      A relatively constant 0 to 7.6 PFU/liter penetrated the membranes used
at average feed water virus concentrations of 9.6 x 10»to 6.8 x 108 PFU/liter.
      Penetration of the membranes by viruses appeared to be a random
phenomenon.

Sproul, O. J. (1972). Virus Inactivation by Water Treatment. J AMER WATER
WORKS ASSN, 64(1):3l-5.

      Nearly  all  water treatment  processes  in  present use  are under
investigation to define better their virus-removal capacity. Recent efforts have
defined some basic mechanisms of virus removal.
      If the virus problem in water supplies is as large as the recent  scientific
and popular literature indicates, then research needs and research efforts stand
in the same relationship as a mountain to a molehill. No research effort at all in
the past 10 years has been given to virus removal by filtration. In some plants,
over  the  same period,  this process has become the only treatment unit,
performing the roles of flocculation chamber, sedimentation tank, and filter.

Stille. W., Kunkel, B..  Merger,  K. (1972). Shellfish-Transmitted Hepatitis.
DTSCH MED WOCHENSCHR, 97(2): 145-7. German.

      Within a period of three years (May 1968 to October 1971) 425 patients
with viral hepatitis were studied at the University Clinic in Frankfurt. In 34, the
disease apparently  followed consumption  of oysters or other shellfish during
travel outside of Germany.

Sweet B. H., Ellender, R. D. (1972). Electro-Osmosis:  A New Technique for
Concentrating  Viruses From Water. WATER RES, 6(7):775-9.

      Concentration of viruses by electroosmosis in a Canalco CF-3 unit was
possible only  in small scale operation (5-liter  quantities). The  technic was
useful with poliovirus 1  inputs  of 0.01 PFU/ml and reovirus inputs of 0.1
PFU/ml or greater only.

      In distilled  water, under optimal  conditions with cellulose acetate
membranes and  flux  rates of 3 ml/cm2/hour,  physical concentrations  of
approximately 40-fold were achieved in 2.5 to  3 hours with up to 80% virus
recovery. Results with tap water were not as good.
      Because  of  the  mechanical  complexity  of the system—its power
requirements, need for osmotic driving solutions  and pumping and refrigeration

                                   20

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systems—the method is not economical for large scale use.  The system may
serve best as a second step procedure following initial concentration of large
volumes of water by either ultrafiltration or other processes. Because it desalts
and dehydrates, the method may also be useful for concentrating viruses from
estuarine waters.

Sweet, B.  H.,  McHale, J. S., Hardy, K. J., Klein, E. (1971). Concentration of
Virus from Water by Electro-Osmosis and Forced-Flow Electrophoresis. PREP
BIOCHEM, 1{1):77-89.

     Poliovirus 1  was  concentrated  from water  in Canalco Model CF-3
Electrophoretic Filter/Concentrator (modifed after Bier) by electroosmosis and
forced-flow electrophoresis.
     Water was removed at a rate up to 0.8 ml/hr/cm* of membrane area by
electroosmosis. With  12-14 V/cm (5-6 amps) and adjusted pumping rates,
20-fold concentration was achieved without virus loss.
     During forced-flow electrophoresis, the virus, negatively charged in an
alkaline buffer, moves toward the positive pole. At 20 V/cm and with adjusted
pumping  rates, the best  concentration achieved  was 3-fold  with  10-fold
dehydration. Virus spill-over at  the cathode and virus adsorption at the anode
were responsible for the poor results, but this may be overcome by adjustment
of the voltage coupled with adjustment of the pumping rates.
     Voltage (30 V/cm) and current (6 amps) have no detrimental effects on
viral stability. These technics appear to be more rapid and gentle than other
methods for concentration of virus and may be  scaled up for practical use.

Tarabcak, M., Kratochvil, I., Milosovicova, A. (1971). Effect of Vaccination
with  Live Poliovirus  Vaccine  on the Grculation of Enteroviruses in  the
Population. J HYG EPIDEMIOL MICROBIOL IMMUNOL, 15(7):258-66.

      Stools from randomly selected healthy  children in East  Slovakia were
tested quarterly in 1959 (before mass vaccination) and from  1960 to 1966
(after  mass  vaccination)  to  determine alterations  in  the circulation of
polioviruses and other enteroviruses in the population. More than 7,300 stools
were tested.
      Since  1963, monthly tests of sewage from nine localities in Kosice and in
the Saca  housing estate have  been  done.. Almost 400 sewage samples  were
tested.
      In 1959 (before mass vaccination), poliovirus detection in the  stools of
healthy children  correspond  to  the mean monthly incidence of poliomyelitis
from 1951 to  1959.
      Since the initial mass vaccinations, no cases of poliomyelitis have been
reported in East Slovakia.  Vaccination of  the  young is carried on routinely.
Circulation of wild polioviruses was suppressed and vaccine strains disappeared
from the population within three to four months after vaccination.

                                   21

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      Mass vaccination with  live poliovaccine  reduced the circulation of
 non-polioviruses at the time  of vaccination and for two  to three  months
 thereafter as compared with 1959.
      The same types of viruses recovered from stools were recovered from
 sewage.
      The circulation  of enteroviruses in a population can be determined by
 detection of these viruses in sewage.
Taylor, F. B. (1972). The Holy Cross Episode. J AMER WATER WORKS
ASSN. 64(4):230-2.

      The series of events is described that led to an outbreak of infectious
hepatitis in the Holy Cross football team after they  consumed water from a
contaminated line.
Venosa, A. D., Ozone as a Water and Wastewater Disinfectant:  A Literature
Review. In "Ozone in Water and Wastewater Treatment", Chapter 5, edited by
F. L Evans  III.  Ann  Arbor Science Publishers, Inc., Ann Arbor, Michigan
(1972), 83-100.

     In a general review of water and wastewater disinfection by ozone, the
inactivation of viruses by this oxidant is discussed.
Victoria,  K.,  Hellstrom, K.-G.,   Rylander,  R. (1972).  Redux  Potential
Measurements for Determining the Disinfecting Power of Chlorinated Water. J
HYG, 70X6):3I3-23.

     In chlorine demand-free water, inactivation of Escherichia coli by sodium
hypochlorite, monochloramine,  dichloramine, halazone, chloramine  T,
cyanuric   acid+sodium  hypochlorite  and  cyanuric acid+monochloramine
correlated  better with  redox potential than with the amount of available
chlorine. For individual  pure chlorine compounds, available chlorine generally
correlated better than the redox potential with bacterial inactivation.
Wachs,  B.  Hygienic Standards  For  Sewage Effluents,  In "Fundamental
Measurements  and  Effluent  Conditions  of Sewerages  in  Canal Stations,
Purification Plants, and Receiving Waters,"  edited by H. Liebman.  Verlag R.
CMdenbourg. Munich, Germany, (1970), 50-78.

      Diseases caused by  viruses and other infectious agents are  discussed,
along with water standards for drinking and swimming.

                                   22

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Wacker, W. E. C., Riordan, J. F., Snodgrass, P. J., Chang, L. W., Morse, L. J.,
O'Brien, T.  F., Reddy,  W. J.  (1972).  The Holy Cross Hepatitis Outbreak.
Clinical and Chemical Abnormalities. ARCH INTERN MED, 130(9):357-60.

     Of 97 men exposed at the same time  to infectious hepatitis virus in
drinking water, one-third developed icteric disease, 60% developed anicteric
disease, and 7% showed no evidence of disease.
     An elevated serum glutamic pyruvic transaminase (SGPT) level was the
most  frequent  chemical  abnormality  in  the  anicteric  group.  Serial
measurements of the levels of SGPT, serum glutamic oxaloacetic transaminase
(SCOT), ornithine carbamyl transferase (OCT), serum alkaline phosphatase,
bilirubin, and cholesterol in the icteric group demonstrated the time course and
extent  of  these abnormalities in  this homogeneous population. The levels of
serum bilirubin, alkaline  phosphatase, and OCT decreased in parallel and were
normal  by 70 days after  exposure. The SGPT and SCOT levels remained
elevated longer.
     By 145 days after exposure, some patients still had elevated SGPT and
SCOT levels. Transient hypercholesterolemia occurred in a significant number
of patients reaching a  maximum 55 days after exposure. The mean Value was
normal 25 days later.
Wallis, C., Henderson, M., Melnick, J. L. (1972). Enterovirus Concentration on
Cellulose Membranes. APPLMICROBIOL, 23(3):476-80.

      Cellulose nitrate membranes adsorb viruses from water in the presence of
salts. Trivalent  salts were more  effective than divalent salts. Thus, 0.5 mM
A1C13 was as effective as 50 mM  MgC 12.
      For testing 500 gallons of water, only 0.24 kg of A1C13 was required in
contrast to 20 kg of MgCl2. Virus was eluted from membranes that had an area
of 486 cm2 with 250 ml of pH 11.5 buffer. Lowering the pH of the eluate and
adding A1C13 permitted the virus to be readsorbed on a 4 cm2 membrane from
which it could be eluted in 1 ml of the high pH buffer.
      This procedure provided the basis for concentrating small amounts of
viruses from large volumes of water.
Wallis, C., Homma, A., Melnick, J. L. (1972). Apparatus for Concentrating
 Viruses from Large Volumes. J AMER WATER WORKS ASSN, 64(3): 189-96.

      An apparatus for recovering viruses from large volumes of water has been
devised which consists of a series of textile filters capable of removing debris
and certain  soluble  materials that interfere with virus recovery, cellulose
membranes or an insoluble polyelectrolyte to which viruses adsorb and from
which they  can be  eluted, pumps,  and other ancillary  equipment. The
apparatus is capable of filtering 300 gallons of water/hour.

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      More than 60% of 100PFU of poliovirus 1 seeded into 100 gallons of tap
water were recovered by this method.
Wallis, C.,  Homma, A., Melnick, J. L (1972)- A Portable Virus Concentrator
for Testing Water in the Field. WATER RES, 6(10): 1249-56.

      A system is described for concentrating viruses from large volumes of
water. The system consists of a water pump, an electric generator, a series of
clarifiers, a virus adsorbent, a virus reconcentrator, a 5- and 1-gallon pressure
vessel with a small tank of nitrogen  as  a source of  positive pressure, and
ancillary equipment, all mounted on 2-wheel carts for easy portability.
      The system was standardized with small amounts of poliovirus. The virus
was added to dechlorinated city tap water in an amount so small it could not be
detected unless the virus was first concentrated.
      In the system, raw tap water containing virus was serially passed through
clarifying filters with porosites of 5 to 1 /im to remove  particulate matter, and
then  through  a  1-p.m  cotton  textile   filter to  electrostatically  remove
submicron  ferric and  other  heavy  metal complexes.   These filters did not
detectably  remove viruses. Salts were then added to the running tap water to
enhance adsorption of the viruses to a fiberglass or cellulose acetate filter. Raw
water was processed at the rate of 33 gph, with complete removal of the virus
from the water. Eighty percent of the virus was eluted from the adsorbent.


Wallis, a, Homma, A., Melnick, J. L. (1972) Concentration and Purification of
Influenza   Virus on  Insoluble Polyelectrolytes.   APPL M1CROBIOL,
23<4): 740-4.

      Influenza virus was concentrated and purified rapidly  by adsorbing it
onto and eluting it from an insoluble poly electrolyte. The influenza virus was
first stabilized at pH 4 to 5, since viruses  adsorb to the polyelectrolyte more
efficiently  in this pH range. A precipitate which forms in influenza harvests
under acid conditions in the cold was removed  by ammonium sulfate at  a
concentration which trapped the  precipitate but  not the virus. Ammonium
sulfate-treated influenza virus in allantoic fluid was readily concentrated on the
polyelectrolyte. Elution yielded a virus concentrate essentially free of nonviral
proteins.

Walton, G.,  Becker, R.  J., Champlin, R. L.,  Faust, S. D.,  McCabe,  L. J.,
Pearson, H. E.. Pogge, F. W.. Weiser, P. W.. Wolf, H. W. (1972). Community
Water Pollution R&D Needs. J AMER WATER WORKS ASSN, 64(4}:211-15.

      In a  general report on research and  development needs, the American
Water Works Association Committee on Pollution Parameters notes the current
state of knowledge on viruses in water, and recommends expanded studies on
the survival and persistance of viruses in that environment.

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Watson, J. T.,  Drewry,  W. A. (1971). Adsorption of f2 Bacteriophage by
Activated  Carbon and Ion Exchange. Water Resources Research Center Report
No. 14, Department of Civil Engineering Research Series No. 11, University of
Tennessee, 238 pages.

     In batch  tests  and in column studies at variable flow rates, activated
carbon  (Pittsburgh and  West  Virginia Nuchar  C-190) and an anion resin
(Amberlite IRA-402) adsorbed seeded coliphage f2 from a synthetic river water
and from  trickling filter effluent. A cation resin (Amberlite IR-122) was not an
effective virus adsorbent. Virus removal could be described by a  first-order
equation.
     On a weight  basis, West  Virginia Nuchar C-190 was the most effective
adsorbent in both the batch tests and in the column experiments. However, in
the columns, Amberlite IRA-402 achieved the  best virus removals in both test
waters when the same depth of adsorbents was used. This removal exceeded
99.9% in both test waters when the flow rate was 0.5 gpm/sq ft. The removals
were considerably greater in the synthetic  river water than in the wastewater
effluent.
     Competition  for adsorption  sites by  other organic matter was a major
factor  in  the wastewater effluent  experiments. Diffusion seemed to play an
important role  in  the virus adsorption process in the synthetic river water
column studies.

Westman,  W.  E. (1972).  Some  Basic Issues  in  Water Pollution Control
Legislation. AM SCIENTIST, 60(11-12):767-73.

     The need for  more  intensive attention to  viruses is considered  in  a
discussion of water pollution control legislation.


Zarma,  M. (1972). Fecal Pollution of Sea Water. MICROBIOL PARAZITOL
EPIDEMIOL, 17(5-6):203-29. Rumanian.

     The survival of viruses in sea water is briefly discussed in a review dealing
primarily with other sea pollution problems.

Zikmund, V., Cech. Ivt,  femin, K., Per man, J. (197 2). Epidemic of Infectious
Hepatitis  in  a School  Caused by  a  Waterbome  Agent.  CS EPIDEMIOL,
21(7): 197-202. Czechoslovakian.

      Early in 1970, an epidemic of infectious hepatitis occurred in a Basic
Nine-Year School in Liberec. The  infection appeared to have been transmitted
by water  from a well supplying the school kitchen. About one-third of those
exposed became ill.
      The water may have been contaminated  by a cesspool from a small house
in which a case of infectious hepatitis had occurred.

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      At least 23 cases of hepatitis were documented, and there may have been
as many as 44.  Tests for Australia antigen on acute and convalescent serums
were negative.


Ziilich, J. A. (1972). Toxicity of Combined Chlorine Residuals to Freshwater
fish. J WATER POLLUT CONTRL FED, 44(2):212-20.

      Laboratory studies showed that quantities of chlorine below measureable
levels added to nontoxic effluents containing thiocyanate produced a toxicant.
The toxicant was probably cyanogen chloride. The threshold concentration of
this compound for rainbow trout appeared to be 0.08 mg/liter.
      Chloramine  concentrations  of  a few tenths mg/liter were lethal to
warmwater fish such as sunfish, bullheads, and minnows.
      Chloramine concentrations of 0.06 to 0.08 mg/liter were lethal to trout.
      Chloramine  concentrations  of  0.085 mg/liter nearly eliminated the
spawning of  the fathead minnow,  and  concentrations of 0.043  mg/liter
significantly reduced their reproduction.
      Chlorinated  effluents were toxic after  dilution to 2 to 4%. Average
concentrations of 0.16 to 0.21 mg/liter residual chlorine caused complete kills
of fathead minnows. As little as 0.07 mg/liter caused a partial kill of the test
fish, and 0.04 to 0.05 mg/liter constituted the threshold concentration in these
wastes.
      An extensive field survey of fish populations in four Maryland streams
showed that chlorinated effluents  significantly reduced  the species diversity
and total number of fish below the outfalls. After the toxic effects had been
eliminated and the organic matter decomposed, the subsequent deoxygenation
of the receiving  streams caused species shifts and many of the more sensitive
fish disappeared.
      Thiosulfate rendered toxic chlorinated compounds nontoxic. This has
been pioven in laboratory and field situations.
                                      aUSGPO: 1973—757-550/1302 Region 5-11


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