United States
Environmental Protection
Agency
Municipal Environmental Research ^
Laboratory                     't
Cincinnati OH 45268
Research and Development
EPA-600/S2-82-024  August 1982
Project Summary
Removal  of Virus from
Public  Water Supplies

J, T. O'Connor, L. Hemphill, and C. D. Reach, Jr.
   Assays of concentrated raw Mis-
 souri River water for the presence of
 naturally occurring human enterovir-
 uses resulted in the recovery of polio-
 viruses, Coxsackie viruses, and ECHO
 viruses. Virus concentrations were
 found to be highest in the winter and
 spring, when river flows (and there-
 fore the effects of dilution) were low-
 est. Statistically significant  inverse
 correlations  were  found between
 virus recoveries and  conventional
 microbiological indicators of water
 contamination (total coliform. fecal
 coliform, and standard plate counts).
   An evaluation of the effectiveness
 of water treatment at a plant in Lex-
 ington,  Missouri, indicated that pre-
 sedimentation plus the addition, of
 lime resulted in the reduction of virus
 levels.  Concentrates derived from
 1.9-m3 (500-gal) samples  of treated
 drinking water were assayed. No iso-
 lates were obtained from the 65 sam-
 ples analyzed.
   Batch  disinfection studies were
 conducted using raw Missouri River
 water treated with chlorine, chloram-
 ine,  chlorine dioxide, or  ozone.
 Though virus levels were very low and
 data were limited, ozone and chlorine
 dioxide appeared to be most capable
 of inactivating viruses under the con-
 ditions employed.
   Studies  of  the effectiveness  of
 physical removal processes were con-
 ducted  using water treatment pilot
 plants constructed of stainless steel
and  housed  in  a  12.2-m  (40-ft)
tractor-trailer. Because raw water vir-
uses  were present  in  insufficient
numbers, bacteriophage-seeded stu-
dies were conducted. Results indi-
cated that a virus reduction of one or
two  orders  of magnitude was
obtained  by physical removal  pro-
cesses alone.  A comparison of con-
ventional  treatment versus direct
filtration  indicated that  comparable
removal could be obtained  by both
processes, although conventional
treatment was somewhat more con-
sistent in performance. Reductions in
•other microbial indicators paralleled
virus removals, indicating the effec-
tiveness of the indicators as monitors
of virus removals.
  The final portion of the study was
conducted by the Capital City Water
Treatment Plant  in Jefferson City,
Missouri.  Chlorine  and  chloramine
were  applied  in  parallel pilot-plant
units for the disinfection of Missouri
River water. Both disinfectants were
capable of producing a finished water
that met  bacteriological standards.
Chloramine, however, proved to be
more stable than chlorine.
  This Project Summary was devel-
oped  by  EPA's Municipal Environ-
mental  Research  Laboratory,
Cincinnati. OH. to announce key find-
ings of the research project that is
fully documented in a separate report
of the same title (see. Project Report
order information at back).

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Introduction
  The efficiency of drinking water treat-
ment processes for the removal and
inactivation of viruses has been studied
extensively.  Because  of the lack  of
methods for detection and enumeration
of naturally occurring viruses at the lev-
els at which they are  present in most
operating treatment plants, virus levels
are  usually  increased by adding
laboratory-propagated  viruses.  The
adequacy of such studies for modellfng
or simulating  actual water treatment
conditions has frequently been questi-
oned. In an attempt to conduct treat-
ment efficiency studies using  only
naturally occurring viruses,  an operat-
ing water  treatment plant using poor
quality source water was identified and
used as the study site.
  Preliminary studies indicated that the
source water contained sufficient natu-
rally occurring viruses  to  make  the
study feasible. Although virus  levels
were not consistently  high throughout
the course of the 3-year study, substan-
tial information on the effects of water
treatment  unit processes on naturally
occurring viruses and microbial indica-
tor organisms was obtained.
Results

Longitudinal Studies of
Virus in  Missouri River
Source Water
  Throughout the study,  naturally
occurring  human enteroviruses were
found in Missouri River water at the
Lexington,  Missouri, water treatment
plant intake. An average of 2.5 viruses
were recovered  in  216 samples of
approximately 0.38 m3 (100 gal) each.
Polioviruses were  most. abundant
(69%), followed by Coxsackie (11%) and
ECHO viruses (5%). Of the total, only
10% of the virus particles were reco-
vered on the 3- to 5-fjm pore size prefil-
ters,  indicating  that  most  of  the
recovered virus particles were notasso-
ciated with the suspended sediment in
Missouri River water.
  Virus concentrations were found to
average  10  plaque-forming  units
(pfu)/0.38 m3 (100 gal) in the winter
and spring of 1977, 1.5 pfu/0.38 m3
(100 gal) in the summer and fall, and 5
pfu/0.38 m3( 100 gal) in the winter artd
spring  of  1978 (Figure 1). If the virus
concentrations  are  normalized  with
respect to river flow, the virus flow in
the Missouri River at Lexington appears
to be more nearly constant throughout
the year. But  seasonal winter  and
spring  virus  flow  peaks appear to
persist.
  Statistically significant inverse corre-
lations were obtained between concen-
trations of virus and microbiological
indicators (fecal coliform, total coliform,
and  standard  plate  counts). These
results, coupled with winter and spring
virus flow peaks, indicate that conven-
tional microbial parameters  may not
always prove  useful  as quantitative
indicators of virus contamination.
Field-Scale Studies of Virus
Removal by Treatment
Processes
  Samples were taken of the presettled,
lime-treated, and finished water at the
Lexington,  Missouri, water treatment
plant over  the  period of December 2,
1976, to March 22,1978. In 21 samples
taken of presented water, a total of 39
viruses were recovered. Seven viruses
were recovered from 14 samples taken
after  adding lime  to the  raw water.
Finally, no viruses were recovered from
the 71 finished water samples (1.9 m3
(500 gal)) collected.
  The results from the survey of plant
performance indicate that the overall
water treatment process at Lexington,
Missouri,  is consistently effective in
reducing viruses to  below  detectable
limits. Moreover, preliminary treatment
of raw Missouri River water with lime
appeared to result  in a measurable
inactivation of the viruses present.
Chemical Disinfection for
Virus Inactivation
  Ozone  and chlorine dioxide both
showed potential for inactivating the
human  enteroviruses present in  raw
Missouri River water when sufficient
dosage and time were provided. Chlo-
rine  and  chloramine,  however,
appeared to be only marginally effective
against viruses when  applied to  raw
water. In general, the reduction of vir-
uses  to below detectable  limits  was
accompanied by near-total destruction
of  fecal and  total  coliforms  and by
reductions in standard plate counts of
two to three orders of magnitude. The
comparatively low numbers of acid-fast
bacteria and yeast found in raw Mis-
souri River water appear to limit their
use as indicator organisms.
Physical Removal of Virus
Particles
  As part of the study of the physics
removal  of virus-sized particles fron
Missouri River water, a mobile wate
treatment research facility was con
structed. Twin water treatment pilo
plants  were built  and mounted in i
tractor-trailer.  A  separate,  limited
edition  internal  report  entitled  Con
struct/on Plans and Operations Manua
for USEPA Mobile Water Treatment
Facility was prepared to provide subse
quent operators of the pilot plant facility
with design calculations,  shop  draw-
ings, fabrication notes, and instructions
for operation, maintenance, and safety.
  For the physical removal studies, bac-
teriophage MS2  was seeded  into raw
Missouri River water as a surrogate for
human enteroviruses  to increase the
number of virus particles present, to
shorten the time between sampling and
obtaining the experimental results, and
to reduce costs.
  Coagulation  with aluminum sulfate
followed by sedimentation and  dual-
media  filtration resulted in an overall
removal  of 93%  of seeded bacterio-
phage.  Comparable  removals   were
obtained with respect to fecal and total
coliforms, fecal streptococci, and stand-
ard plate count organisms.
  In other studies on removal of  natu-
rally occurring MS2  bacteriophages,
coagulation with a cationic polyelectro-
lyte followed by direct filtration resulted
in an  87%  removal of seeded  MS2
bacteriophage.
  With  direct  filtration, results  were
more erratic than those  obtained with
conventional   treatment.  Again, the
removal of other indicator organisms
paralleled  the  removal of   MS2
bacteriophage.
  Turbidity reductions reflected  orga-
nism   removals.  Influent  turbidities
ranged from 19  to 180  NTU. Effluent
turbidities ranged from 1 to 7 NTU for
conventional treatment. Direct filtration
produced turbidities in the range of 0.4
to 8.5  NTU, except for  a  single high
value of 77 NTU. This turbidity break-
through occurred  during  a period of
high influent turbidity.
  Overall,  one-  to  two- order-of-
magnitude removal of seeded virus and
other microbial indicators  of pollution
was obtainable by both conventional
water  treatment  and direct filtration
under  proper  operating  conditions.
Direct  filtration, however,  was  more
subject to upset during periods of high
influent turbidities.

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Figure 1.     Virus in raw Missouri River water, December 1976 through December 1978..



                                                                              3
1978
                                                                                 iu.oovDM«MrnwnNaomce: MB -559-017/0792

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   Chlorine Versus Chloramine
   for Water Disinfection
     Studies of organism removal and dis-
   infection by chlorine and chloramine at
   Jefferson City, Missouri, began with a
   comparison  of organism  reductions
   with  and  without the  addition of
   chlorine.
     Aluminum  sulfate coagulation  and
   sedimentation removed 80% to 85% of
   the standard plate  count organisms.
   With  filtration, organism  removal
   increased to  90%. Adding  chlorine
   increased  organism  reductions to
   99.9% or more.
     The subsequent comparison of chlo-
   rine versus  chloramine (each  added
   before the rapid mix tank) indicated that
   both disinfectants  were  effective in
   attaining bacteriological drinking water
   standards. Moreover, chloramine  pro-
   duced  an order of magnitude fewer
   trihalomethanes.
     With  the  inclusion   of GAC post-
   adsorbers in the pilot-plant operation at
   Jefferson City, Missouri, it was possible
   to observe the removal of trihalome-
   thanes as well as the reduction in total
   organic carbon and substances  that
   have trihalomethane-forming potential.
     Comparable disinfection  could  be
   achieved with chlorine and chloramine,
   but  effluent  trihalomethanes were
   lower with chloramine than  with the
   use of both chlorine and GAC.

   Summary and Conclusions
     The results of the virus removal stu-
   dies indicate  that both sedimentation
   processes using coagulants and various
   types  of filtration  processes usually
   remove more than  90% of the viruses
   present in the water.  Results of the
        pilot- and field-scale studies tended to
        be similar to those of the laboratory-
        scale studies. Process control is the
        primary  factor determining the  effi-
        ciency of removal processes. The varia-
        bility in  removal  efficiencies  of the
        coagulation-flocculation  and filtration
        processes points up the need for main-
        taining adequate disinfection  in addi-
        tion to the  removal  processes. The
        efficiency with  which  turbidity  is
        removed by sedimentation and filtration
        generally indicates the efficiency of
        virus removal.

        Other Reports Based on This
        Research
          The following publications are based
        on research conducted under this grant
        and have been published or are in
        preparation.
          Reach,  Jr., C.  D., Hemphill,  L, Akin,
          E., and O'Connor, J. T. "Naturally-
          Occurring  Human Enteroviruses in
          Missouri River Water." (In prepara-
          tion;  to  be  submitted  to Water
          Research.)
          Reach, Jr., C. D., Hoff, J. C., Logsdon,
          G., and O'Connor,  J. T. "Chemical
  Inactivation and Physical Removal ol
  Naturally-Occurring Human Entero-
  virus in Missouri  River Water." (In
  preparation; to be submitted to Jour-
  nal American Water Works Assoc.)
  Reach, Jr., C. D., Hoff, J. C., Logsdon,
  G., and O'Connor, J. T. "Chlorine ver-
  sus Chloramine for the Disinfection
  of Missouri River Water." (In prepara-
  tion; to be  submitted to Journal
  American Water Works Assoc.)
  Reach,  C.  D., O'Connor, J. T.,  and
  Hamphill,  L.  'Virus  and  Bacterial
  Quality  of Missouri  River." Proc.
  Annual Meeting of American Water
  Works Association. Part I.  pp. 519-
  529. 1979.
  Reed, G. D., Reach, Jr., C. D.,  and
  O'Connor,  J.T.   "Trihalomethane
  Precursor Removal under Two Modes
  of  Disinfection." Journal  American
  Water Works Association (accepted
  for publication).
  The full report was submitted in ful-
fillment of Grant No. R-804903-02 by
the University  of  Missouri-Columbia
under the sponsorship of the U.S. Envir-
onmental Protection Agency.
          J. T. O'Connor. L. Hemphill. and C. D. Reach, Jr. are with the University of
            Missouri. Columbia, MO 65211.
          John C. Hoff is the EPA Project Officer (see below).
          The complete report, entitled "Removal of Virus from Public Water Supplies,"
            (Order No. PB 82-230 327; Cost: $15.00, subject to change) will be
            available only from:
                  National Technical Information Service
                  5285 Port Royal Road
                  Springfield.  VA 22161
                  Telephone: 703-487-4650
          The EPA Project Officer can be contacted at:
                  Municipal Environmental Research Laboratory
                  U.S. Environmental Protection Agency
                  Cincinnati, OH 45268
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
               Postage and
               Fees Paid
               Environmental
               Protection
               Agency
               EPA 335
Official Business
Penalty for Private Use $300

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