United States
 Environmental Protection
 Agency
 Health Effects Research
 Laboratory
 Cincinnati OH 45268
 Research and Development
 EPA-600/S1-81-029 Apr. 1981
 Project Summary
Mutagenic  Activity and
Chemical  Characterization  for
the  Palo  Alto  Wastewater
Reclamation  and  Groundwater
Injection  Facility

P. L. McCarty, J. Kissel, T. Everhart, R. C. Cooper,  and C. Leong
  At the Palo Alto Reclamation Plant
0.044 m3/s  (1 mgd) of secondary
effluent is reclaimed through a series
of wastewater treatment processes
including high  lime treatment,
aerator-fountain  spraying,  single-
stage recarbonation, ozonation,
mixed-media   filtration,  activated-
carbon adsorption, final chlorination,
and  storage. A  portion  of  the
reclaimed water  is injected into a
series of  wells to serve as  a barrier
against the intrusion of sea water from
San Francisco Bay.
  Mutagenic  activity (by the Ames
test)  was consistently found to be
present  in  the  secondary treated
municipal wastewater influent to the
Reclamation Facility. This activity was
not reduced significantly by high lime
treatment, air stripping, recarbona-
tion, or ozonation, even though these
processes did remove a portion of the
overall organic content of the waste-
waters and  many  of the volatile
organic  compounds. Activated-
carbon adsorption was effective in
removing mutagenic activity to such a
degree that mutagenic activity could
not be found  in water used for injec-
tion or that taken from monitoring
wells. Chlorination resulted in  an
increase  in  mutagenic activity. A
laboratory study  demonstrated  that
this increase in  activity would not
result if chlorine dioxide rather than
chlorine was used for disinfection.
  The extracts used for mutagenic
analyses contained a broad range of
fatty acids, phthalates, aromatic com-
pounds,  and  several  unidentified
bromine-containing compounds.
  This Project Summary was develop-
ed by EPA's Health Effects Research
Laboratory, Cincinnati, OH, to an-
nounce  key findings of the research
project that is fully documented in a
separate report of the same title (see
Project Report ordering information at
back).


Discussion
  Water  reclamation and reuse are be-
coming more important as increasing
pollution and water usage reduces the
supply of clean water available for bene-
ficial uses. Advanced wastewater treat-
ment processes have been developed to
protect receiving waters and to reclaim
wastewater for a variety of purposes.
  The nature of wastewater is depend-
ent upon the types of  industrial and
municipal wastes  introduced into the
sewerage system Wastewaters may
contain a variety of toxic compounds,
some of  which may not  be altered or
removed during treatment, and some

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 may be  formed during the treatment
 process itself.
  The  direct use of  reclaimed waste-
 waters as part of a municipal drinking
 water supply is not currently permitted.
 Before this is allowed, the concern over
 the potentially harmful effects of con-
 taminants remaining in the water must
 be alleviated. Although research aimed
 at the  identification and quantification
 of trace materials in water constitutes
 an active area of environmental chemi-
 stry, little is known aboutthe long-term,
 or chronic, effects of exposing a popula-
 tion to trace levels of these dissolved
 impurities  over a time period of many
 years.  There is a reasonable probability
•that many of these compounds are car-
 cinogenfc  Epidemiological studies do
 exist which tend to  show a degree of
 correlation  between  the  increased
 incidence of several forms of cancer and
 polluted  surface  waters   serving as
 potable water sources, as well as to
 chlorinated drinking  water.
  Several   reports   document the
 presence of  mutagenic substances in
 drinking  water as well as reclaimed
 wastewater These investigations relied
 chiefly upon a bacterial mutagenicity
 assay,  the  Ames  test, to detect the
 presence of mutagens. There is some
 evidence that many  compounds which
 are bacterial mutagens, as determined
 by the Ames test, are also carcinogenic
 to animals. Thus, this relatively simple
 test may provide some indication of the
 presence of potentially harmful sub-
 stances in  water.
  This  study  on   the  presence  of
 bacterial mutagens in reclaimed waste-
 water at Palo Alto, California, was insti-
 gated  as a result  of a previous study
 evaluating   the  presence  of   such
 mutagens in samples taken from sixdif-
 ferent  California municipal wastewater
 treatment  facilities.  For  two  of the
 plants, including the one at Palo Alto, it
 was reported that the mutagenic activity
 was intensified during the course of the
 treatment process  In the study reported
 here, the mutagenic  activity at the Palo
 Alto Reclamation Plant was evaluated
 in more  detail on  samples taken after
 various stages of treatment so that their
 effect  in either producing or reducing
 mutagenic  activity  could be   better
 evaluated.  In  addition,  chemical
 analyses were conducted on collective
 parameters, such  as chemical oxygen
 demand, total organic carbon, and total
 organic halogens, as well as on a wide
 range of specific organic compounds, in
order to determine if  any correlation
with mutagenic activity existed
  Special studies were conducted on
the formation of  mutagemcally active
compounds   during  the  chlormation
process, and a comparison was made
with  an alternative disinfectant,
chlorine dioxide. In addition, an evalua-
tion was made  of  the  reduction  in
mutagenic activity during passage  of
the reclaimed water through an aquifer
following groundwater injection.


Conclusions
  Positive mutagenic activity to strains
TA98  and TA1535 with S9 activation
was  consistently  measured  in  the
secondary treated wastewater influent
to the Reclamation Facility. Chlorination
of  the   secondary  treated  influent
resulted  in an increase in mutagenic
activity,  especially to strain TA1 535.
  Laboratory  disinfection studies con-
firmed that mutagenic activity  of the
secondary   treated   wastewater
increased following  Chlorination,  but
would not result  had chlorine dioxide
disinfection  been  used.  Subsequent
processes of high lime treatment, strip-
ping, recarbonation, and ozonation had
little effect in reducing mutagenic activ-
ity,  although  in  combination  these
processes were effective in removing
many volatile organic  constituents  of
health concern.
  Activated-carbon adsorption  proved
very effective in  reducing mutagenic
activity  as well,  as  in reducing  the
organic   content  of  wastewater  as
measured by  the collective parameters
of chemical oxygen demand (COD), total
organic  carbon (TOG), and total organic
halogens (TOX). Because of the effec-
tiveness of activated-carbon adsorption
in  reducing  mutagenic  activity,  few
samples of the reclaimed wastewater
used  for  groundwater injection con-
tamed  positive  mutagenic  activity.
Positive  mutagenic  activity was  not
found in any samples  taken from the
groundwater  monitoring wells.
  An  average of only about 12 percent
of the organic material present in the
samples analyzed for mutagenic activity
was  contained  in  the XAD-acetone
extract   concentrates.  Fatty  acids,
phthalates,  several  aromatic
compounds,   and  several  unidentified
bromine-containing compounds were
present  in the XAD-acetone extracts
used  for mutagenic analysis.
  The Palo  Alto  Reclamation  Facility
was proven  effective in  reducing the
concentration of a wide range of specific
organic compounds and  mutagenic
activity  contained  in the  secondary
treated  influent  wastewater. Several
materials  remain in the reclamation
plant effluent at low concentration, and
mutagenic  activity  was  detected in
some samples.


Results and
Recommendations
  The efficiency of the individual treat-
ment processes for  removal (or forma-
tion) of a wide range of specific organic
compounds and group parameters was
measured  and are summarized in this
report  Chlorination  results  in the pro-
duction of a significant concentration of
non-purgeable, but  otherwise uniden-
tified,  chlorinated organic compounds,
as well as trihalomethanes. Formation
or removal of such compounds did not
seem  to  correlate  with changes in
mutagenic activity Air stripping by the
fountain-spray  system  was   most
effective in  removing volatile organic
compounds, while activated carbon was
responsible for  removals obtained for
most other organic materials.            t
  Future  work  should  be  directed
towards evaluating further the organic
material remaining m reclamation plant
effluent,  as  most  of this  material
remains uncharactenzed. In addition,
the nature of compounds or mixtures of
compounds responsible for  mutagenic
activity should be determined. The use
of the Ames mutagenic analysis for
judging the  health  risk of reclaimed
wastewater  should  be  further
evaluated

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P L. McCarty, J. Kissel, andT. Everhartare with the Department of Civil Engineering,
  Stanford University, Stanford, CA 94305,  andR. C. Cooper and C. Leong are with
  the School of Public Health,  University of California. Berkeley, CA 94720.
Herbert R. Pahren is the EPA  Project Officer (see below).
The complete report, entitled "Mutagenic Activity and Chemical Characterization
  for the Palo Alto Wastewater Reclamation and Groundwater Injection Facility,"
  /Order No PB 81 -179 590, Cost: $9.50. 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:
        Health Effects Research Laboratory
        U S.  Environmental Protection Agency
        Cincinnati,  OH 45268
  > US 30VERNMENT PRINTING OFFICE 1961 757-012/7066

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United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
                                                                                                            Postage and
                                                                                                            Fees Paid
                                                                                                            Environmental
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Official Business
Penalty for Private Use $300
              0000J?9

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