f/EPA
                                      United States
                                      Environmental Protection
                                      Agency
                                   Health Effects Research
                                   Laboratory
                                   Cincinnati OH 45268
                                      Research and Development
                                   EPA-600/S1-81-002  Mar. 1981
Project  Summary
                                      Health  Risks  of  Human
                                      Exposure  to  Wastewater
                                      C. S. Clark, C. C. Linnemann, Jr., G. L Van Meer, G. M. Schiff, P. S. Gartside,
                                      A. B. Bjornson, E. J. Cleary, J. P. Phair, C. R. Buncher, D. L. Alexander, S. E.
                                      Trimble, and B. C. Barnett
I
  The  primary  objective  of  this
research was to determine the health
effects associated with occupational
exposure to biological agents present
in municipal  wastewater. An addi-
tional objective was to determine the
sensitivity  of  the methodology for
detecting potential health impacts of
other wastewater exposures, such as
recreational contact  with surface
water receiving wastewater effluents.
The procedure was a prospective sero-
epidemiologic study  applied  to
municipal wastewater workers and
controls in three metropolitan areas:
Cincinnati, Ohio; Chicago, Illinois;
and  Memphis,  Tennessee.  The
primary  study  group  consisted  of
more than  100 workers recruited at
the time they began work at activated
sludge plants and who remained in the
study for a minimum of 12 months. In
addition, a Chicago group of 30 expe-
rienced  sewage  treatment  plant
workers were  included   and,  in
Cincinnati, two  other wastewater-
exposed groups were recruited con-
si sting  of  about  50 sewer
maintenance workers and 50 primary
wastewater treatment plant workers.
The latter group was recruited into the
study just  before start-up  of plant
improvements that included activated
sludge facilities.  The  purpose  of
including this group was to differen-
tiate between  aerosol exposure and
exposure to wastewater and sludge
through  those  operations associated
with primary wastewater treatment.
  The protocol involved quarterly col-
lection of  blood,  throat, and rectal
swabs; yearly medical examinations;
collection of illness information; work
observations;  and  environmental
monitoring. Initial  recruitment  of
workers  began  April 1975  in
Cincinnati; July 1976 in Chicago; and
July 1977 in Memphis. Final speci-
mens in all  cities were collected in the
fall of  1978. The  serological survey
included testing for  antibodies to a
large group of viruses and bacteria and
determination   of   immunoglobulin
levels. Work observations were used
to evaluate the level of the workers'
contact with wastewater and, in con-
junction with the biological air moni-
toring, to assess the extent of contact
with  aerosols.  The  environmental
monitoring included viral and bacterial
analyses of wastewater and the use of
six-stage Andersen samplers to deter-
mine the respirable concentrations of
bacteria in the work areas of the plant.
  A total  of over 500  volunteers
participated in  the.study including
both subjects and controls.

  This Project Summary was devel-
oped by EPA's Health  Effects  Re-
search Laboratory, Cincinnati, OH, to
announce 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).

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Introduction
Objectives
  The  purpose of this  study  was to
determine the health effects, if any, of
the  occupational  exposure   to  the
viruses, bacteria, and parasites present
in municipal wastewater. The central
feature was an extensive surveillance
of  the health  of  100  newly-hired
activated  sludge  treatment plant
workers during  a  minimum of  12
months of  occupational exposure to
wastewater. Specific objectives of the
project were:

1.   To  determine whether wastewater
    workers develop clinical illness or
    specific  bacterial, viral, and para-
    sitic infections due to occupational
    exposure to sewage

2.   To  determine  the immunologic  re-
    sponse among workers presumed
    to be exposed to  a high level of anti-
    genic   simulation,   i e.,   through
    wastewaters.

3.   To  determine  whether wastewater
    workers serve  as  a reservoir of
    certain  infections   and,   if   so,
    whether members of the workers'
    families are affected.

4   To  determine the effect of exposure
    10  aerosols generated by the acti-
    vated sludge treatment process.

5   To  determine  the concentration of
    bacterial  aerosols at wastewater
    treatment  plants, and to compare
    them to levels at other public works
    facilities.

  An underlying objective of the study
was the determination of the sensitivity
of various elements of  the epidemio-
logical-serological  approach   for   the
detection of wastewater-related health
effects Such a  determination would
permit an assessment of the  potential
application of this methodology to  the
study  of health risks associated with
other  population  exposures to waste-
waters.  The  latter  would   include
persons working with  or living near
wastewater   land-disposal  facilities,
persons living in  the vicinity of waste-
water  treatment  plants, and  persons
engager ~i recreational use of bodies of
water  receiving waste effluents.
Background
  Information  regarding  the  human
health  risks associated with contact
with wastewater and related materials
brought  about by  occupational  and
other exposures is  limited.  However,
assumptions concerning these risks are
providing  motivation   for   the
promulgation  of  state and  federal
standards  designed to protect
populations from  various wastewater


Table 1.     List of Viruses, Bacteria,
            and Immunoglobulins for
            Which   Workers'   Sera
            Were   Tested  for Anti-
            bodies.
Viruses
1.  Polio 1, 2, and 3
2.  Coxsackie A-7, A-9,  A-16,  A-21,
    and B-1  to B-6
3.  Echo 1, 3, 4, 5, 6. 8, 9,  11, 13, 14,
    19, 24, 30
4.  Reov/rus
5.  Adenovirus
6.  Cytomega/ovirus
7.  Herpes simplex
8.  Hepatitis A antibody and Hepatitis B
    antibody and antigen
      Test Methods: M/croneutra/iza-
      tion for Polio, Coxsackie and all
      Echo   viruses;   Radioimmuno-
      assay for Hepatitis A  and B; and
      Complement  Fixation  for  the
      others.

Bacteria
1.  Salmonella: Group A, B, C, D, andE
      Test Methods' Rapid slide  agglu-
      tination (1975-1976)
      Microagglutination (1978)
2.  Leptospira
      Test Method: Microagglutination
3.  Legionella pneumophila
      Test Method: Indirect immuno-
      fluorescence

Immunologic Factors
1.  IgA
2.  IgG
3.  IgM
4.  Rheumatoid Factor
       Test Methods: Radial immunodif-
      fusion   for  immunoglobulins,
      latex  reaction  for  rheumatoid
      factor
exposures. The growing  emphasis on
the land application of wastewater and
sludges as a viable method of wastes
utilization  increases  the  need  for
reliable and up-to-date information on
the  health  risks,  if  any, involved.
Wastewater  also  contains  a  wide
variety of harmful chemicals which may
under some conditions compromise the
health  of  wastewater  workers.
However,  chemical hazards were not
considered in this study.
Methods
  The study consisted of an intensive
serologic survey correlated with epide-
miological, clinical, and environmental
data on  the study populations.  The
central feature of the design was an
evaluation of the effects of occupational
exposure to wastewater over at least a
12-month   period   based   on  the
measurement  of specific  viral  and
bacterial antibodies and immunoglobu-
lin  in  sera  collected  over that time
period  In order  to  help  separate the
effect of the occupational exposure from
that associated  with other  possible
disease pathways, appropriate control
groups were utilized. In addition, many  I
of  the  wastewater  workers  were
recruited into the study at the time of
their initial employment m the waste-
water treatment industry
  Each quarter blood  samples, throat
swabs,   and  fecal  samples  were
collected from participants in the study.
Blood  specimens were  used for the
serologic surveys and the throat swabs
and fecal specimens were used for
analysis  for  bacteria,   viruses  and
parasites. (Parasitic examinations were
performed only during the early period
of the study in Cincinnati.)The serologic
survey involved  the determination of
antibodies to a group of viruses  and
bacteria  and  measurement  of
immunoglobulm  levels  (Table  1). Of
concern  in  the  serologic survey  was
whether the prevalence  and level of
antibodies   were   different   in  the
wastewater exposed  and  control
groups,and  whether  the number of
infections as indicated by increases in
antibody concentration  was  different
among the various study  groups.
   Illness information  was obtained  by
 monthly health  diaries  maintained  by
 the workers supplemented by telephone
 and on-the-job contact. Illness symptom
 information  from  all   sources  wasj

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 combined in a mannerdesignedtoavoid
 double counting and was categorized as
 "respiratory,"  "gastrointestinal,"  and
 "other."  The data  processing proce-
 dures  provided   for  coding  various
 possible  combinations of these  The
 definition of  these illness categories
 was based on the symptoms indicated
 on the health diary, as follows.

 Respiratory      -  Cold  symptoms,
                   sore  throat,  cough
                   or other lower  res-
                   piratory symptoms.

 Gastrointestinal  -  Nausea,  vomiting,
                   diarrhea, or intesti-
                   nal upset

 Other           -  Fever,   persistent
                   headache, eye  in-
                   flammation or other
                   eye  trouble,   ear-
                   ache or other  ear
                   trouble,  skin infec-
                   tion,  rash, boils, or
                   open sores.
   From time  to  time  job  locations of
 participants were visited to determine
 types and levels  of exposure and other
 work conditions. Work site  air  was
 monitored  periodically  for bacterial
 concentrations,   and  concurrent
 wastewater samples at the wastewater
 treatment  plants  were analyzed  for
 bacterial and viral concentrations. Six-
 stage Andersen samplers were use'J to
 collect the airborne bacteria. In order to
 process  the  air samples  as soon as
 possible, preparation  and analyses of
 the plates for bacterial sampling were
 performed  in  Chicago  by  the
 Metropolitan  Sanitary  District of
 Greater Chicago (MSDGC), in Memphis
 by the Memphis State University, and
 by the bacteriologist on the study  staff
 in Cincinnati. All wastewater samples
 for virus assay were processed by the
 aluminum hydroxide  continuous  flow
 centrifuge technique and assayed by a
 plaque assay procedure.
   On a  yearly basis,  study volunteers
 were offered a medical evaluation that
  included  hematology, urmalysis, pul-
  monary  function testing,  blood chem-
  istry (including tests of liver and kidney
 function), and examination by a physi-
 cian.
   A  number   of  comparisons   and
 correlations  were  made  from   the
 epidemiological, environmental,
t clinical,  and  serological aspects of the
 study.
  As initially conceived, the study goals
were  limited to the first three specific
objectives listed above  The population
groups  planned for  the  study were
sewer  and   highway  maintenance
workers  in  Cincinnati.  The research
design initially called for recruitment of
an equal number of menbegmning their
respective jobs as those recruited who
had two or more years experience,  for
both exposed and control groups. Soon
after the study was initiated economic
conditions in the municipal government
forced a moratorium on  hiring new
employees  in  the  Cincinnati  Public
Works Department which eliminated
prospects for   establishing  a  newly
employed highway maintenance study
group.
  About  one year  after this research
began,  its goals  were expanded to
include a determination of the  health
effects associated with the dispersion of
aerosols generated by  the activated
sludge wastewater treatment process
(Specific Objectives 4 and 5).  At this
time the study design was  expanded to
include two additional exposed popula-
tion groups: 50 men at  the Cincinnati
Mill  Creek Sewage  Treatment Plant
which was  in the process of being
expanded from primary  wastewater
treatment  to   include  the  activated
sludge process; and a total of 100 men
newly employed  at  activated  sludge
treatment plants. Since  only one-third
of the  100  inexperienced  activated
sludge plant workers were expected to
be available in Cincinnati, thestudy was
expanded to  include  workers  at the
plants in Chicago and Memphis. These
cities were chosen because at least 50
new employees would be  hired  within
the next year, the plants contained the
activated process in open  basins with
porous plate-type diffusers, they were
reasonably accesible to Cincinnati, and
the   plant  administrators  were
agreeable to the study. MSDGC hired a
significant  number of  new  workers
because of normal work force turnover
by virtue of its  size. In addition, its long
experience in   wastewater treatment
and   the  technical   support  for
monitoring available from its Research
and Development Department made it
very suitable for study. Memphis was in
the final stages of construction of  its
second  activated  sludge treatment
plant, the North Treatment Plant,  an
entirely new facility. In addition, its first
plant,  the  Maxson  Wastewater
Treatment Plant had plans  to hire more
workers.
   Comparison  groups  in  Cincinnati,
 Chicago and Memphis  were highway
 maintenance  water  treatment  plant
 workers and  gas and  electric  utility
 workers, respectively.
 Conclusions
 1.   Gastrointestinal illness rates were
     higher in the inexperienced waste-
     water exposed workers than in the
     experienced workers and controls,
     Wastewater   workers  were  not
     found  to  be  subject  to   any
     detectable risks due to parasites
     present  in wastewater. There was
     only slight evidence, if any, to sug-
     gest that there were risks due to
     viruses and bacteria in wastewater.

 2.   Immunoglobulm levels  were  not
     found to be consistently higher in
     wastewater-exposed workers than
     in  controls in any of  the cities
     studied.

 3.   Wastewater   workers  were  not
     found to serve as a source of viral
     infections for their family members.

 4.   In a few instances levels of antibody
     to  certain viruses  appeared to be
     related   to  level of  exposure to
     wastewater aerosols.

' 5.   Bacterial aerosol levels in buildings
     where   wastewater  sludge  was
     being processed were  generally
     higher than levels adjacent to out-
     door  aeration tanks at  the same
     treatment plants.

 6.   Since the seroepidemiologic  ap-
     proach   did   not   detect  any
     significant health effects of occu-
     pational  exposure to wastewater, it
     is unlikely that this approach would
     detect potential  health impacts in
     populations with lower  levels of
     exposure to wastewater.


 Recommendations
   Since the  basic design of this study
 was  to   compare  antibody  levels
 between  two quarters in each year of
 the study, all sera collected over the
 entire  study period have  not been
 tested. It is  possible that changes in
 antibody  levels were not detected by
 semiannual testing. Selected viruses of
 high prevalence should be studied by
 testing all sera for each individual.
                                                                                   > US GOVERNMENT PRINTING OFFICE 1M1-757-02I/70J2

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      Because  the  predominance  of
    seroconversions to Hepatitis B were in
    the sewage-exposed groups (seven of
    123  sewage-exposed  workers  com-
    pared  to one of 52 control workers),
    additional  testing should be done on
    workers not yet evaluated to determine
    whether this trend persists.
      Additional testing of sera of workers
    occupationally exposed to soil, such as
    some sewer and highway maintenance
    workers, would be useful m attempting
    to verify previous suggestions that soil
    exposure increases the risk of infection
    to Legionella pneumophila
      Because of the limited testing of sera
    for antibody to  Hepatitis A, the initial
    sera of a number of workers has not
    been tested. It would be useful to test
    these sera to determine whether any of
    these  people acquired  the  antibody
    during the course of the  study
       Serologic  testing  for  rotavirus or
    parvovirus  agent was not performed
    during  the  study. Since  these  viruses
     are now recognized as a  major cause of
     gastrointestinal  illness,  the  sera col-
     lected  during  the  study should be
     analyzed for antibody  to  them.
            C. S. Clark, C. C. Lmnemann, Jr., G. L.  Van Meer, G. M. Schiff, P. S. Gartside.
              A. B. Bjornson,  E. J. deary, J.  P. Phair, C. R. Buncher, D. L. Alexander,
              S. E. Trimble, and B C. Barnett are with the University of Cincinnati Medical
              Center, Cincinnati, OH 45267.
            Walter Jakubowski is the EPA Project  Officer (see  below).
            The complete report,  entitled "Health  Risks of Human Exposure to Waste-
              water," /Order No. PB 81-143 406; Cost: $15.50, subject to change) will be
              available only from:
                    National Technical Information Service
                    5285 Port Royal Road
                    Spring field, 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
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
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Fees Paid
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Penalty for Private Use $300
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