GREAT  LAKES  WATER QUALITY
        Status of  Pertinent  EPA  Researc
                              December  197

        National  Environmental  Research Center
        Corvallis,  Oregon

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                                 EPA-660/9-74-002
                                        Report to
                   INTERNATIONAL JOINT COMMISSION
                         United States and Canada
                       GREAT LAKES WATER QUALITY
                 Status of Pertinent EPA Research
                                    December 1974
                                          Authors
                                    A. F. Bartsch
                                 Charles L. Frank
                                 Spencer Peterson

National Environmental Research Center/ Corvallis
    200 S.W. 35th Street/Corvallis, Oregon 97330

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This report was prepared under the auspices of the
Great Lakes Research Advisory Board.  The Board was
created as a result of the United States/Canadian
Great Lakes Water Quality Agreement of 1972.  The
agreement commits both countries to the improve-
ment of the Great Lakes water quality and requires
that pollution abatement measures be either com-
pleted or initiated by December 31, 1975.  The
eight-member board, principal scientific advisor
to the International Joint Commission—United
States and Canada and to the Great Lakes Water
Quality Board, was organized to assure that
research objectives of the Commission would be
accomplished efficiently.  Dr. A. F. Bartsch,
Director of the U. S. Environmental Protection
Agency's National Environmental Research Center
at Corvallis, Oregon, serves as United States
co-chairman of the Great Lakes Research Advisory
Board.

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                                 CONTENTS

                      Introduction                  1

                      Nitrilotriacetate             2

                      Viruses                       6

                      Disinfection of Wastewater    9

                      Fish Contamination           13

                      References                   16
Published by the Office of Public Affairs,
National Environmental Research Center,
Corvallis, Oregon, U. S. Environmental
Protection Agency.  This document is
available to the public through the
National Technical Information Service,
Springfield, Virginia 22151.

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                                   INTRODUCTION

This report summarizes major segments of research
activities the U.S. Environmental Protection Agency
has completed or now has under way that relate to
nitrilotriacetate (NTA), viruses, disinfection of
municipal wastewater and the effects of polychlori-
nated biphenals (PCBs) on fish and fish products.

The summary is intended to highlight research areas
which may have particular impact on objectives of
the International Joint Commission as recommended
in its 1973 annual report and should not be con-
sidered inclusive of all activities presently
funded by EPA in the above subject areas.

During FY 1975, EPA plans to expend about $2 million
for continued research on:

     • Nitrilotriacetate—$300,000
     • Viruses—$600,000
     • Disinfection of municipal wastewater—$830,000
     • Freshwater effects of PCBs on fish—$300,000

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                            NITRILDTRIACETATE
Eutrophication
Research has been conducted at the Pacific Northwest
Environmental Research Laboratory, Corvallis, Oregon,
to determine the influences of NTA on eutrophication.
Results indicate that in general NTA has no dramatic
effects on algal growth rates.  Any stimulatory or
inhibitory effects appear to be related to the che-
lating properties of NTA and interaction with algal
trace metal metabolism.  There was no.reduction in
the NTA content of test medium when subjected to
nitrogen-limited 21-day algal assays, indicating
that NTA did not serve as a nitrogen source.  Algal
growth stimulation from phosphorus and secondary
sewage effluents was much greater than any resulting
from NTA.

Metals from river  sediments near  industrial  and
mining outfalls were  solubilized  by NTA more
readily  than those  from  lake  sediments.   Copper,
iron, manganese, cadmium, and  zinc were solubi-
lized from sediments  in  the presence  of NTA.  How-
ever, copper from highly reduced  lake  sediments
was not  readily solubilized from  Detroit  River
sediments.

In summary,  NTA appeared to have  no dramatic stimu-
latory or  inhibitory  effects on freshwater eutrophi-
cation processes  (National Eutrophication Research
Program, 1970).  This  is in contrast with the effects
of NTA on  marine estuarine systems where  small
amounts  of NTA may  produce a  trace metals deficiency,
thus limiting  productivity  (Erickson, Maloney, and
Gentile, 1970).
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The long term effects of NTA in marine waters, how-
ever, might produce just the opposite impact.  Since
marine bacteria are capable of metabolizing NTA, it
is possible that the end products of NTA degradation
could serve as a nitrogen source,  thus stimulating
algal blooms in marine systems which are usually
nitrogen limited.

Degradation Products and Energy Transfer Effects

Laboratory studies at the Robert S. Kerr Environ-
mental Research Laboratory evaluated the probable
impact of nitrilotriacetic acid on ground water.
A report (Dunlap, Cosby, McNabb, Bledsoe, Scalf,
1973) indicated that the potential for pollution
of ground water by NTA from detergents involved:

       • possible introduction into ground water
         of undesirable organic degradation com-
         pounds of NTA, and

       • possible transport of heavy metals into
         and through ground water by NTA in re-
         charge water.

It was concluded that further clarification of
these possibilities was necessary.  Thus, studies
of NTA degradation in anaerobic ground water
environments were initiated in 1973.

Anaerobic ground water environment work is near-
ing completion.  Results permit the following
tentative conclusions:

     • degradation of NTA in anaerobic sub-
       surface environments is not likely to
       result in accumulation of significant
       quantities of undesirable organic de-
       gradation products in ground water,  and

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     • the rate of NTA degradation in anaerobic
       ground waters will vary widely depending
       on microbial populations present and
       nutrient availability.

However, prediction of probable NTA degradation rates
in specific anaerobic subsurface environments is not
feasible at present since little information is avail-
able on these subsurface environments and their asso-
ciated microbial communities.

Further work on NTA degradation by bacteria and yeasts
in the marine environment is in progress at the Gulf
Breeze Environmental Research Laboratory in Florida.
Since this project is in its initial.stage, consider-
able work remains to be done, but preliminary evi-
dence indicates that there is some degradation of NTA
in estuarine systems by the indigenous microbes.  This
has been evidenced through the evolution of "  C label-
ed C0? from   C labeled NTA molecules.

Following the correction of minor problems with isotope
analysis, a survey of the Pensacola Bay System will de-
termine the differences in NTA degrading bacteria and
yeast biomasses in polluted and non-polluted areas.

Toxicity

A grant awarded to the Academy of Natural Sciences
of Philadelphia (Ruth Patrick, project officer) is
designed to determine the effects of various concen-
trations of potential micronutrients, particularly
NTA, on the efficiency of energy transfers and the
structure of aquatic communities in artificial
streams.  Growth rates and biomass accumulations of
various algal species are being studied and the
efficiency of organics decomposition by bacteria
and fungi in the presence of NTA is being determined.

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To date gas chromotographic procedures for NTA
analysis have been refined to the point of detect-
ing 0.02 mg/1, a level at the lower end of the
range expected in natural habitats.  NTA added to
test streams at a maintained concentration of
0.2 mg/1 induced no alteration of algal species
composition or primary productivity over the
control communities.  The utilization of dis-
solved organic substances by heterotrophs in the
system was not impaired.

The toxicity of NTA to various organisms in the
aquatic food web has been tested at the National
Water Quality Laboratory, Duluth, Minnesota.
Experiments with Daphnia magna have shown that a
strong negative correlation exists between water
hardness and NTA toxicity (Biesinger, Andrew,
and Arthur, 1974).  That is, NTA was more toxic
in soft water than in hard water.

Bioassay experiments of NTA, with copper and
zinc added, confirmed earlier reports that che-
lates of copper and zinc with NTA are relatively
nontoxic.  This was also true for the amphipod
Gammarus pseudolimnaeus and the fathead minnow
Pimephales promelas (Arthur, Lemke, Mattson,
and Halligan, 1974).  The chronic no-effect
level of NTA to the amphipods was 19 mg/1 and
54 mg/1 to the fathead minnows.  This work has
been completed and published.

Current year (FY-75) expenditures on NTA research
amount to $200,000.  Projected expenditures for
FY-76 total $60,000.

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                              VIRUSES

Identification

EPA's National Environmental Research Center,
Cincinnati, Ohio, is presently conducting re-
search with waterborne viruses and their pos-
sible health hazards.  During FY-75, work is
proceeding in developing methods for concen-
tration, recovery, and identification of vi-
ruses from water.  Specifically, a grant has
been let that will initiate development of an
immunochemical method for identification and
detection of waterborne viruses.  Another
project has been initiated to develop methods
for identification and detection of viruses
infecting blue-green algae.

The water supply program conducts health effects
research concerned with the microbiological con-
taminants of water supplies.   Work has been done
to improve detection methods for viruses in raw
and finished water.  An integral part of this re-
search is the design and implementation of epi-
demiological studies of contaminants that are
suspected to be transmitted by drinking water.
The water supply program is concerned with the
improvement of virus detection methods and has
determined that reliability in detecting enteric
viruses appears to be greatly improved by con-
centrating the organisms from 400 or more
liters of water.  Emphasis is placed on threa
basic concentration methods that show promise:

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     • the flow-through gauze pad sampler technique,
     • the membrane virus adsorption technique, and
     • the insoluble polyelectrolyte (PE 60) method.

Tests are being conducted simultaneously under expe-
rimental conditions using poliovirus type 1 and con-
centration techniques with 400 liters or more of
test water.  A number of bench type experiments are
planned to optimize and/or refine many of the
procedural steps.

A study is under way to determine if viruses can be
detected in finished drinking water and to evaluate
and compare, under actual field conditions, the
efficiency of the three virus concentration tech-
niques.  The study will relate the presence or
absence of viruses in finished drinking water to
treatment processes, water source and protection,
total and fecal coliform densities, standard plate
counts, and zoomicrobe populations.
Epidemiology

An integral part of water supply research is the de-
sign and implementation of epidemiological studies
of contaminants whose transmission by drinking water
is suspected but not definitely proven.  Two types
of studies are under way:

     • an intensive literature search relating to
       the minimal infective dose of enteroviruses
       and data collection on recovery of poliovirus
       from stools, sewage, streams, etc., and

     • an objective determination of the age that
       children become infected with enteroviruses.

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The water supply program maintains a tabulation on
waterborne disease outbreaks.  During 1971-72, 47
waterborne outbreaks resulting in 6,817 illnesses
were reported in the United States.  This is con-
tinuous research.

The air pollution health effects program at the
National Environmental Research Center at Research
Triangle Park, North Carolina, has in progress an
epidemiological study to look at the effects of
airborne biological contamination from sewage
treatment plants.  The study will focus on the
relationship of airborne contaminants and the
increased incidents of health problems in human
populations living near municipal sewage treatment
plants.

This is the second year of a three-year study.  No
results have been reported.  If the findings indi-
cate a correlation between health problems in
areas surrounding treatment plants, additional
work will be initiated to determine specific agents
responsible for the health problems.

Prior to FY-74, $703,000 was spent on this health
effects research.  In FY-74, $300,000 was expended.
For FY-75, $600,000 is budgeted, with another
$600,000 planned for FY-76.

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                         DISINFECTION OF WASTEWATER

Treatment Plant Effluent

The Advanced Waste Treatment Laboratory of
the National Environmental Research Center in
Cincinnati, Ohio, has completed and is currently
involved in a number of projects relating to the
disinfection of wastewater effluents.  'These
studies can be .classified into two major cate-
gories:

     • effects of disinfection on aquatic organisms,
       and
     • developments and demonstration of associated
       technology for effluent disinfection.
In—depth projects are scheduled to examine the
state-of-the-art of these disinfectants:

     • chlorine, hypochlorites, and chloramines,
       with dechlorination by sulfur dioxide,
       activated carbon, or bisulfite;
       bromine, bromamines, and bromine chloride;
       iodine;
       ozone;
       ultraviolet light; and
       chlorine dioxide.

Each disinfectant will be reviewed with regard to
its chemistry in aqueous solutions.  The bacteri-
cidal, virucidal, and cysticidal nature of each
species will be compared, as well as its toxicity.

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Various external factors which impact the effec-
tiveness of the disinfectant will be examined.
Analytical methodology—such as principles of
operation, chemistry, on-site monitoring of dosage
and residual control—will be covered.  Engineering
and design aspects—including method of application,
mixing chambers, contact chambers, safety measures
and operation and maintenance—will be covered.

In the first phase of a demonstration project under
way in Wyoming, Michigan, parallel disinfecting
techniques for effluents from an activated sludge
wastewater treatment plant are being evaluated for
disinfection efficiency and residual toxicity.  The
processes under study include chlorination, chlori-
nation   with dechlorination, ozonation, and bromine
chloride treatment.

An untreated fifth stream serves as a non-disinfected
control.  Results, which are preliminary and applicable
to the waste discharged at this particular plant,
indicate that the minimum bacteriological quality stan-
dard of 1,000 total coliforms and 200 fecal coliforms
per 100 ml can be easily attained with bromine chloride
and chlorine with and without dechlorination at residual
disinfectant concentrations of less than 3.0 mg/1 and a
contact time of 30 minutes.  At this particular plant,
the ozone disinfection efficiency has not been this
high.

The acute fish bioassay data indicate that chlorine is
indeed toxic to various fish species, but that its
toxicity is removed upon dechlorination with sulfur
dioxide.  Bromine chloride is also toxic, but to a
lesser degree.  The evaluation of ozone for possible
toxicity is under way.  Chronic fish bioassay deter-
minations are in process to investigate the long-term
effects of the disinfection processes on various fish
species.
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This demonstration project represents the first time a
comprehensive parallel comparision of various disin-
fectants and subsequent fish toxicity studies have been
conducted under field conditions.  The final results
could have substantial impact on EPA policy regarding
standards which will be published for wastewater effluent
The second phase of this project, just getting under
way, will follow the same format with effluent from a
trickling filter plant.  (Note: A workshop presenting
research results from this project was held in October
1974, in Wyoming, Michigan.)


Storm Water

A number of projects have been initiated to examine
the various aspects of the disinfection of storm
water.  These studies have concentrated on techniques
to obtain quick dosage of the wastewater through the
use of varioirs baffling and mixing techniques.
Several combinations of disinfectants in conjunction
with these physical techniques have been examined.
As a result of these studies, rapid mixing and high
turbulence have resulted in disinfection to the 1,000
total coliform per 100 ml level, with 1-minute con-
tact time and 5 mg/1 chlorine dose.

In the near future, several other projects will
demonstrate dechlorination procedures and flash
mixing and contact systems.  An evaluation of dis-
infection necessary for preparation of wastewater
for ground water recharge is planned.

The total funding level for projects specifically
related to disinfection prior to 1TY-75 amounted to
$2.1 million.  Corresponding figures for FY-75 and
FY-76 are $630,000 and $800,000.  Many other pro-
jects examine various aspects of disinfection in a
peripheral fashion.
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Disinfectant Toxicity

Concurrently with the above demonstration program,
additional effects studies are being conducted at
the National Water Quality Laboratory to determine:

     • safe concentrations of chloramines to
       Pacific salmon, trout, and crayfish;
     • effects of chlorinated sewage on biota
       of Milwaukee Harbor and adjacent Lake
       Michigan;
     • effects of chlorinated power plant con-
       denser cooling water on rainbow trout
       and fathead minnows;
     • toxicity of chlorinated municipal wastes
       to aquatic life;
     • chlorination and ozonation products of
       municipal sewage;
     • acute and chronic toxicity of chlorinated
       secondary sewage.

Results of .the effects research have shown that
chlorinated sanitary effluents are extremely
toxic to confined aquatic organisms, especially
fish.  Most fish will avoid areas of higher con-
centrations when the discharge is continuous.

National Water Quality Laboratory criteria
recommendations are:

     • 0.002 mg/1 total residual chlorine to
       protect salmonids and other more sensi-
       tive groups, and
     • 0.01 mg/1 total residual chlorine to
       protect warmwater fish and other less
       sensitive groups.

Through FY-74, EPA has expended $886,000 for in-
house work relating to the disinfection of muni-
cipal sewage and its effect on aquatic organisms.
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                              FISH roiOTlINATION

EPA has completed a number of reports (National
Water Quality Laboratory, 1973) that specify the
effects of PCBs on freshwater organisms and aquatic
ecosystems, including information on toxicity
tissue residues and possible hazards to man.  Data
generated can be used to determine pollution stan-
dards for water, supplemental information for reg-
istration, and improving evaluation methods.

Continuous-flow and static bioassay tests have been
conducted with several PCBs.  Fish reproduction as a
measure of relative toxicity has been determined for
Arochlor 1221 (A1221), 1232, 1242, 1248, 1254, 1260,
1262, and 1268.  Studies are continuing on the acute
and chronic toxicity of various Aroclors to fish and
invertebrates.

Two 9-month continuous-flow bioassays and several
intermediate length continuous-flow tests were con-
ducted to determine safe levels of Aroclor 1242, 1248,
and 1254 for the fathead minnow Pimephales promelas
and Aroclor 1248 for the flagfish Jordanella floridae.
Calculated 96-hour LC50 values for newly-hatched fat-
head minnows were 7.7 yg/1 for Aroclor 1254 and 15
yg/1 for 1242.  Three-month-old fatheads had a 96-
hour LC50 of 300 yg/1 for 1242.  Reproduction occurred
at and below 1.8 yg/1 1254 and at and below 5.4 yg/1
1242.  Newly-hatched young were the most sensitive
life stage.  Growth of young fatheads was affected
above 2.2 Ug/1 1248.  None survived above 5.1 yg/1
after 30 days.  Young flagfish did not survive at
1248 concentrations above 5.1 yg/1 and did not grow
well above 2.2 yg/1.
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Fathead minnow fry less than 24-hours-old at the start
of the test were exposed for 30 days to Aroclors 1248
and 1260.  Median toxicity values of 4.35 yg/1 for 1248
and 2.5 yg/1 for 1260 were obtained.  Total mortality
was observed at 8.0 yg/1 and 7.0 yg/1, respectively.
Chronic tests have recently been completed with these
Aroclors at concentrations of 3 yg/1 and below.  While
the data have not been analyzed statistically, there
were no readily apparent effects on survival, growth,
or reproduction.

The fathead minnows exposed to Aroclor 1248 and 1260
accumulated about 80 percent of the residue present
after the 250-day exposure within the first 90 days.
Aroclor 1248 was accumulated in,.lipids up to concen-
trations approximately 1.3 x 10  times that in water.
Previous work at the National Water Quality Laboratory
measured bioaccumulation factors for Aroclor 1242 and
1254.  The data from these PCB studies indicate a con-
sistent direct relationship betweeen the percentage of
chlorine in the PCB and the bioaccumulation factor
(inverse relationship with water solubility.)

Female fathead minnows in both exposures contained
about twice as many PCBs on a wet weight basis as
the males and showed a greater standard deviation.
Males exposed to 3.5 yg/1 Aroclor 1248 accumulated
194 + 12 yg/gm of the PCB on a wet weight basis.
Since these variations can be accounted for largely
by the greater percentage of lipids in the females,
the data show that the residues cannot be adequately
described throughout the bioassay unless the sex of
the fish is included in the analysis.
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The results of this research indicate that Aroclors
are very toxic.  Chronically safe levels for fish
and invertebrates are in the range of 1-5 pg/1.
Resultant residues are unacceptably high, and a
much lower concentration near 1 mg/1 will be nec-
essary to limit residue levels for the protection
of the consumer—man or wildlife.

During the EPA Toxic Substances Hearings conducted
in May 1974, the following items on the current use
of PCBs were obtained during cross-examination of
industry witnesses:

     • admission that the production of Aroclor
       1254 has not diminished since 1969—
       20,000 tons/year;

     • typical industries using PCBs discharge
       20 to 25 Ibs/day into the environment;  and

     • retraction of industry contention that
       PCBs containing four or more chlorines
       are readily degraded in the environment.
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Arthur, J. W., Lemke, A. E., Mattson, V. R., and
Halligan, B. J.  1974.  Toxicity of sodium
nitrilotriacetate (NTA) to the fathead minnow
and an amphipod in soft water.  Water Res.
8:187-193.

Biesinger, K. E., Andrew, R. W., and Arthur, J. W.
1974.  Chronic toxicity to NTA (nitrilptriacetate)
and metal-NTA complexes to Daphnia magna. Jour.
Fish. Res. Bd. Can. 31:486-490.

Dunlap, W. J., Cosby, R. L., McNabb, J. F., Bledsoe,
B. E., and Scalf, M. R.  1971.  Investigations Con-
cerning Probable Impact of Nitrilotriacetic Acid
on Ground Waters.  EPA Water Poll. Cont. Research
Series 16060 GHR 11/71.

Erickson, S. J., Maloney, T. E., and Gentile, J. H.
1970.  Effects of nitrilotriacetic acid on the
growth and metabolism of estuarine phytoplankton.
Jour. Water Poll. Cont. Fed. 42(8):R329-335.

National Eutrophication Research Program.  1970.
Field and laboratory investigation to evaluate
the influence of nitrilotriacetate (NTA) on
eutrophication. Report to the Commissioner
of Federal Water Quality Administration.  Unpub.
rpt.

National Water Quality Laboratory.  1973.  Summary
of Research Findings for Fiscal Year 1973.  U. S.
EnvJ
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