PB81-222566
Reduction of Toxicity to Aquatic Organisms by
Industrial Wastewater Treatment
(U.S.) EG and G, Bionomics
Wareham, MA
Prepared for
Environmental Protection Agency
Cincinnati, OH
Jul 81
U.S. DEPARTMENT OF COMMERCE
National Technical Information Service
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EPA 600/3-81-043
REDUCTION OF TOXICITY TO AQUATIC
"ORGAN ISMS~B Y~INDUSTRrAITWAS TEWATE R~ TREATMENT"
by
George A. Gary and Michael E. Barrows
EG&G, Bionomics
790 Main Street
Warehanrr Massachusetts— 0'2571
Contract #68-03-2631
William Horning, II
Newtown Fish Toxicity Station
U.S. Environmental Protection Agency
3411 Church Street
Cincinnati, Ohio 45244
U.S. Environmental Protection Agency
Cincinnati, Ohio 45244
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NOTICE
THIS DOCUMENT HAS BEEN REPRODUCED
FROM THE BEST COPY FURNISHED US BY
THE SPONSORING AGENCY. ALTHOUGH IT
IS RECOGNIZED THAT CERTAIN PORTIONS
ARE ILLEGIBLE, IT IS BEING RELEASED
IN THE INTEREST OF MAKING AVAILABLE
AS MUCH INFORMATION AS POSSIBLE.
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TECHNICAL REPORT DATA
{Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-81-043
2.
ORD Report
3. RECIPIENT'S ACCESSION NO.
222366
4. TITLE AND SUBTITLE
Reduction of Toxicity to Aquatic Organisms by
Industrial Wastewater Treatment
5. REPORT DATE
June 1981
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
George A. Gary, and Michael E. Barrows
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
EG&G Bionomics
790 Main Street
Wareham, Massachusetts 02571
10. PROGRAM CLEMENT NO.
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency
Newtown Fish Toxicology Station
3A11 Church Street
Cincinnati, Ohio 45244
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
15. SUPPLEMENTARY NOTES
16. ABSTRACT
The specific goal of this research was to conduct 24-hour static acute bloassays with "untreated"
Influent and "treated" effluent using fathead minnows (Plmephales promelaa) and water flea (Daphnia magna) to
biologically evaluate the effectiveness of industrial wastewater facilities in reducing effluent toxlclty to
aquatic organisms. Of primary Interest to the EPA was an evaluation of the capacity of the wastewater
treatment facilities of the pesticide industry for reducing toxicity. To accomplish the stated goal, on-site
24-hour static acute toxicity tests were_jifii|£rmed during ten consecutive workdays at seven industrial sites.
Five__of__t]ie_test sites are defined as (pestitldji> manufacturers, while the remaining sites consisted of an
ofganft-ehemlEM. manufacturer and a bleached^kraf t (^peroy3r. The effectiveness of the treatment plants was
determined by performing static acute toxicity tests wiEhthe fathead minnow (Plmephales promelaa) and the
water flea (Daphnia magna) on samples of the wastewater collected before and after treatment. Results of the
studies are expressed in terms of both median lethal concentrations (LCSO's) as Z effluent and lethal units.
The data from these studies indicate that the wastevater treatment plants provided an average efficiency
of 98Z in reducing the toxicity of "untreated" wastewaters. Neither species tested proved to be a more
sensitive indicator of toxicity, though a larger data base is required to make valid appraisal.
Of Interest was the observation that while some wastewater treatment facilities provide good efficiency
(98+Z) in reducing toxicity, the resulting effluent still represented a relatively high number of lethal
units. This was a result of the fact that the "untreated" influent entering the waste treatment system
contained a very high level of lethal units and a subsequent 98Z reduction of that level still resulted in a
toxic wastewater.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
c. COSATI Field/Group
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisRepon)
UNCLASSIFIED
21. i'O. OF PAGES
20. SECURITY CLASS (This page I
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (Rev. 4-77) PREVIOUS EDITION is OBSOLETE
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-DISCLAIMER-
This report has been reviewed by the EG&G, Bionomics Labora-
tory, U.S. Environmental "Protection Agency, and approved for
publication. Approval does not signify that the contents neces-
sarily reflect the views and policies of the U.S. Environmental
Protection Agency, nor does mention of trade names or commercial
products constitute endorsement or recommendation for use.
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EXECUTIVE SUMMARY
On-site, 24-hour static acute toxicity tests were performed
during 10 consecutive work days at 7 industrial sites to deter-
mine the effectiveness of the respective wastewater treatment
facilities in reducing toxicity of the discharges to aquatic
organisms. Five of the test sites are defined as pesticide manu-
facturers, while the remaining sites consisted on an organo-
chemical manufacturer and a bleached-kraft paper mill. The effec-
tiveness of the treatment plants was determined by performing
static acute toxicity tests with the fathead minnow (Pimephales
promelas) and the water flea (Daphnia magna), on samples of the
wastewater collected before and after treatment. Results of the
studies are expressed in terms of both median lethal concentra-
tions (LCSO's) as % effluent and lethal units.
The data from these studies indicate that the-wastewater
treatment plants provided an average efficiency of 98% in re-
ducing the toxicity of untreated wastewaters. Neither species
tested proved to be a more sensitive indicator of toxicity,
though a larger data base is required to make valid appraisal.
Of interest was the observation that while some wastewater
treatment facilities provide good efficiency (98+%) in reducing
toxicity, the resulting effluent still represented a relatively
high number of lethal units. This was a result of the fact that
the untreated influent entering the waste treatment system con-
tained a very high level of lethal units and a subsequent 98%
reduction of that level still resulted in a toxic wastewater.
iii
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CONTENTS
Executive Summary '. iii
Figures v
Tables . vi
Acknowledgements xi
1. Introduction 1
2. Materials and Methods 3
Dilution Water 3
Wastewater Sampling. . 3
.-_ Test" Organisms". 77 77. 77 77. 77 ."7". 77.7". 7777". 77 .""7.7". 4
Fathead minnow 4
Water flea ', 5
On-Site Test Procedures 6
. General 6
Fathead Minnow Tests 7
Daphnia magna Tests. ., 8
Ames Test '. 8
Statistics ..-..« 9
3. Results . 10
Union Carbide, South Charleston, West Virginia. 10
Monsanto, Muscatine, Iowa 12
Mobay Chemical, Kansas City, Missouri* ....• 14
Monsanto, Luling, Louisiana 15
Diamond-Shamrock, Green Bayou, Texas 17
E.I. duPont de Nemours, LaPorte, Texas 18
International Paper, Georgetown, South Carolina 19
4. Discussion 22
5. References .• 25
6. Tables 26
Appendix. 57
IV
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FIGURES
Number Page
1 Flow diagram for wastewater treatment plant at
South Charleston, West Virginia (Union Carbide
effluent) . 11
2 Flow diagram for waste treatment facility at
Monsanto, Muscatine, Iowa 13
i
-~ .4-3- —Flow-diagram for—waste-treatment •fctcility~atr
Monsanto, Luling, Louisiana 16
!
4 Flow diagram for waste treatment facility at
International Paper, Georgetown, South Carolina.... 21
v
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Number
List of the seven industrial wastewater treatment
facilities which were chosen for on-site effluent
toxicity studies, „ 26
LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "untreated influent" and
"treated effluent" from the South Charleston, West
-Virginia industri~al~wastewa~ter treatment" p~lan~t~~.7~. ~27
Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the South Charleston, West Virginia in-
dustrial wastewater treatment plant 28
Results; of water chemistry analyses performed during
the on-site testing of fathead minnows to "untreated
influent" and "treated effluent" from the South
Charleston, West Virginia industrial wastewater
treatment plant. These values represent the range
of measurements observed during the ten consecutive
days of on-site testing.. 29
Results, of water chemistry analyses performed during
the on-site testing of Daphnia magna to "untreated
influent" and "treated effluent" from the South
Charleston, West Virginia industrial wastewater
treatment plant. These values represent the range
of measurements observed during the five days of
testing 30
LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "untreated influent" and
"treated effluent" from the Monsanto, Muscatine,
Iowa industrial wastewater treatment plant. 31
Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the Monsanto,. Muscatine, Iowa industrial
wastewater treatment plant 32 -
VI
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Number \ Page.
8 Results of water chemistry analyses performed during
the on-site testing of fathead minnows to "untreated
influent" .and "treated ef .fluent" .from the -Monsanto,
_ .Hoscatine:, Iowa industrial 'sasitewater treatment
~ 'plant,. :33iese yalqes xepr-esent tfae range o:f
_
men'tslofislirvecl during tne ten consecutive days of
testing .............. . . . ............................ 33
9 Results of water chemistry analyses performed during
the on-site testing of Daphnia magna to "untreated
influent" and "treated effluent" from the Monsanto,
Muscatine, Iowa industrial wastewater treatment
plant. These values represent the range of measure-
ments observed during the five days of testing ...... 34
10 LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "treated effluent" from
- --- ~the Mobay Chemical 7~K~ansas~Ci~ty ~ Mi's sour i~iridus trial ~ •'"-*•
wastewater treatment plant ............... ........... 35
11 Results of water chemistry analyses conducted on 100%
test water during ten consecutive days of on-site
testing at the Mobay Chemical, Kansas City, Missouri
industrial wastewater treatment plant .............. 36
12 Results of water chemistry analyses performed during
on-site testing of fathead minnows and Daphnia magna
to "treated effluent" from the Mobay Chemical,
Kansas City, Missouri industrial wastewater treat-
ment plant. These values represent the range of
measurements observed during the testing period ..... . 37
13 LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "untreated influent" and
"treated effluent" from the Monsanto, Luling,
Louisiana industrial wastewater treatment plant ..... 38
14 Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the Monsanto,, Luling, Louisiana indus-
trial wastewater treatment plant .................... 39
15 Results of water chemistry analyses performed during
the on-site testing of. fathead minnow to "untreated
influent" and "treated effluent" from the Monsanto,
Luling, Louisiana industrial wastewater treatment
plant. These values represent the range of measure-
ments observed during the ten consecutive days of
on-site testing ...................................... 40—
Vii.
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JSumber ; Page
16 Results of water chemistry analyses performed during
the testing of Daphnia magna to "untreated influent"
7 and "treated effluent" from the Monsanto, Luling,
Louisiana industrial wastewater treatment plant.
r These values^ represent the range of meagu
observed during the five days of on-site testing..,«. 41
17r LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "untreated influent" and
"treated effluent" from the Diamond Shamrock, Green
Bayou, Texas industrial wastewater treatment plant.. 42
18 Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the Diamond-Shamrock, Green Bayou, Texas
industrial wastewater treatment plant 43
19 Results of watef~c"heinistry~analys"es^peFformed^uri'ng" "*
the on-site testing of fathead minnows to "untreated
influent" and "treated effluent" from the Diamond
Shamrock, Green Bayou, Texas industrial wastewater
treatment plant. These values represent the range
of measurements observed during the ten consecutive
days of on-site testing 44
20 Results of water chemistry analyses performed during
the on-site testing of Daphnia magna to "untreated
influent" and "treated effluent" from the Diamond
Shamrock, Green Bayou, Texas industrial wastewater
treatment plant. These values represent the range
of measurements observed during the five days of
on-site testing 45
21 LC50 values and lethal units for fathead minnows ex-
posed to two "treated influents" and a "treated
effluent" from the duPont, LaPorte, Texas industrial
wastewater treatment plant 46
22 LC50 values and lethal units for Daphnia magna exposed
to two "treated influents" and a "treated effluent"
from the duPont, LaPorte, Texas industrial waste-
water treatment plant 47
23 Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the duPont, LaPorte, Texas industrial
wastewater treatment plant 48
viii
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23 ;»esul*3S ,-o'f -water cfaemis-txy aaaiyses per£o:nnea ;&er±ng
the on-site testing of fathead minnows to two
"treated influents" from the duPont^ LaPorte, Texas
industrial izastewater treatment plant- IThese values _
.represent the range of jneasurements observed Coring _
tfie~t.en consecutive days~o:E on^sXte tes.ting....«......... 4~9
25 Results of water chemistry analyses performed during
the on-site testing of fathead minnows to "treated
effluent" from the duPont, LaPorte, Texas industrial
wastewater treatment plant. These values represent
the range of measurements observed during the ten
consecutive days of on-site testing 50
26 Results of water chemistry analyses performed during
the on-site testing of Daphnia magna to two "treated
influents" from the duPont, Laporte, Texas industrial
.-.. wastewater treatment "plants These~~values~repre"seht
the range of measurements observed during the five
days of on-site testing 51
27 Results of water chemistry analyses performed during
the "on-site testing of Daphnia magna to "treated
effluent" from the duPont, LaPorte, Texas industrial
wastewater treatment plant. These values represent
the range of measurements observed during the five
days of on-site testing.. 52
28 LC50 values and lethal units for fathead minnows and
Daphnia magna exposed to "untreated influent and
"treated effluent" from the International Paper,
Georgetown, South Carolina industrial wastewater
treatment plant 53
29 Results of water chemistry analyses conducted on 100%
test waters during ten consecutive days of on-site
testing at the International Paper, Georgetown,
South Carolina industrial wastewater treatment plant 54
30 Results of water chemistry analyses performed during
the on-site testing of fathead minnows to "untreated
influent" and "treated effluent" from the Interna-
tional Paper, Georgetown, South Carolina industrial
wastewater treatment plant. These values represent
the range of measurements.observed during the ten
consecutive days of on-site testing 55
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Mumber • Page
33. 3ssaal-ts of water chemisl^ry .analyses parfoxsaed
during the on-site testing of Daphnia magna
to "untreated influent" and "treated effluent"
from the International Paper, Georgetown, South
... Carolina industrial_wastewater treatment_p_lant_._
"THese values~fepresent the range of measurements
observed during the five days of on-site testing...56
x
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ACKNOWLEDGEMENTS
We would like to express our gratitude and sincere appre-
ciation to David Elwood and Brian Robinson, the very competent
biologists who performed in the capacity of principal investi-
gators and also as the primary personnel in charge of the
mobile laboratory facility throughout the majority of this
study program. We would also like to extend our appreciation
to all support personnel from EG&G, Bionomics and EG&G, Mason
Research Institute who participated in this project.
Additionally, the authors would also like to express their
sincere appreciation to each of the companies participating in
this study for the extensive time and effort they devoted in
supporting the requirements of Bionomics' mobile bioassay unit
and laboratory personnel.
XI
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SECTION 1
INTRODUCTION
In 1977, the United States Environmental Protection Agency
was charged by Congress under the Federal Water Pollution Con-
trol Act Amendments (PL-95-217) to develop a program which would
protect aquatic life by eliminating the discharge into the
nation's waterways of toxic pollutants in toxic amounts. Under
contract to the EPA (#68-03-2631), EG&G, Bionomics1 personnel
performed on-site toxicity testing at seven industrial waste-
water treatment facilities throughout the United States. The
data generated from these studies will aid the Agency in devel-
oping effluent discharge guidelines related to the toxicity to
aquatic organisms of industrial wastewater streams.
The specific goal of this research was to conduct 24-hour
static acute bioassays with ."untreated" influent and treated
effluent using fathead minnows (Pimephales promelas) and water
flea (Daphnia magna) to biologically evaluate the effectiveness
of industrial wastewater facilities in reducing effluent toxici-
ty to aquatic organisms.
To accomplish this task, it was necessary to evaluate the
toxicity of the wastewaters before and after biological and/or
chemical treatment procedures. Results of the acute toxicity
studies are expressed in both the conventional format as median
lethal concentrations (LCSO's) and as lethal units, a concept
which is similar to that of toxic units as described by Esvelt,
Kaufman and Selleck (1971). The lethal unit concept as it
applies to effluents, is the concentration of -the effluent divi-
ded by the 24-hour LC'5'0. This unitless expression of toxicity
may be ideally suited for use with wastewater discharges of di-
versified effluents and can be incorporated into the regulatory
process of controlling and limiting toxic discharges.
Of primary interest to the EPA was an evaluation of the
capacity of the wastewater treatment facilities of the pesticide
industry for reducing toxicity. The Agency's rational in selec-
ting this industry was based on the premise that discharges from
pesticide manufacturers might be expected to be relatively toxic
and perhaps not readily degraded even during biological treat- .
ment. The industries which participated in this study were
Monsanto Company, Muscatine, Iowa; Mobay Chemical Company, Kansas
City, Missouri; Monsanto Company, Luling, Louisiana; and Diamond
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Shamrock Corporation, Green Bayou,. Texas. Most chemical manu-
facturing complexes are not limited to the production of pesti-
cides alone. These sites were selected because they had segre-
gated pesticide wastewater treatment facilities which do not (for
the most part) receive wastewater from other manufacturing pro-
cesses and because the majority of the treatment systems were
candidate best available technology economically achievable (BAT)
systems or portions thereof, la addition to the above 4 pesti-
ci£s manufacturers, studies s?ers conducted with a camples organo-
chemical effluent at Union Carbide Corporation, South Charleston,
West Virginia, a complex organo-pesticide effluent at E.I. duPont
Nemours & Company, Inc., LaPorte, Texas and a bleached-kraft
paper mill effluent from International Paper Company, Georgetown,
South Carolina. Table 1 presents the sites in order visited
along with dates and other pertinent information.
Contained in this report is a description of the test pro-
cedures and results of the biological and chemical findings at
each site.
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SECTION 2
MKTERISLS ;KND TJETBODS
On-site testing was carried out at each site in one of
EG&G, Bionomics" Mobile Aquatic Toxicology Laboratories. Unless
otherwise stated, procedures used in conducting the on-site tests
followed "Methods for acute toxicity tests with fish, macroin-
vertebrates, and amphibians" (EPA, 1975). All raw data generated
from these studies are permanently stored in the archives at
EG&G, Bionomics, Wareham, Massachusetts.
Dilution Water
The standard diluent and control water used throughout this
contract was a constant quality reconstituted "hard water" pre-
pared according to EPA (1975) methodology. A potable water '.
supply was used to prepare the reconstituted water at each site.
The potable water was first filtered through an activated carbon
cylinder and then through mixed bed deionizers which resulted in
dechlorinated deionized water with a conductivity of less than
1 micromho per centimeter (iamho/cm) . The specified amounts of
the reagent-grade chemicals were then added to this water to
prepare batches of 1000 liters (£) each of the reconstituted
diluent.
Each batch of reconstituted water was then mixed for at
least 1 hour in order to allow for complete dissolution of the
chemical salts. Mixing was accomplished by intensive aeration
using a 2odarR aeration system. The resulting water was charac-
terized by having a total hardness of 160-180 mg/£ as calcium
carbonate (CaC03), a total alkalinity of 110-120 mg/i as CaC03,
a pH of 7.6-8.0, and a specific conductivity of 400-600 ymhos/cm.
Prior to use, the temperature of the reconstituted water was
adjusted, as necessary, to 22°C by use of either a thermostati-
cally controlled ThermoquartzR (MT3-905), 9000 watt stainless
steel immersion heater or 1 H.P. FridgidR Unit chiller (epoxy
coated) equipped with a self-contained thermostat.
Wastewater Sampling
The specific design of the wastewater treatment plants
varied from one site to the next depending on the nature and
composition of the wastewater being treated. Wastewater samples,
which were generally representative of the major contributory
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pesticide wastewater stream entering the treatment plant {"un-
treated" influent) and the final discharge stream from the waste-
water treatment plant ("treated" effluent), were collected at
each site.
""' Depending on the site, 1-2 "untreated" influent wastewater
streams were tested. In certain instances, direct access to an
incoming wastewater stream was not possible until after it had
already entered some portion of the treatment .system (e.g.,, grit
basin or pH neutralization basin). At these sites the influent
samples were obtained immediately prior to the next step in the
treatment process. The complexity of some wastewater treatment
plants (especially E.I. DuPont, LaPorte, Texas) made it impos-
sible to test influent samples which were wholly representative
of -the total pesticide wastewater load entering the treatment
system. This was a result of multiple wastewater streams enter-
ing the treatment plant at different stages in the treatment
process.
"Treated" effluent samples were obtained at sampling points
located immediately after the last step in the treatment process.
Since many of the sites have multiple wastewater treatment
systems which are utilized for treating other manufacturing pro-
cesses, the "treated" effluent tested in these studies represent-
ed only the wastewater discharged directly from the pesticide
wastewater treatment system. These samples were collected prior
to dilution with other process wastewater streams or non-contact
cooling waters and therefore are not representative of the com-
pany's total discharge entering the receiving stream.
Test Organisms
Fathead minnow—Fathead minnows (Pimephales promelas) used
in these studies were raised from eggs in the culture facilities
of EG&G, Bionomics. These fish were cultured in water which was
a mixture of Bionomics1 well water and Town of Wareham non-
chlorinated, non-treated well water. The culture water is char-
acterized by having a range of total hardness and total alkalin-
ity of 25-39 mg/Jt and 20-39 mg/Jl as CaCOs, respectively, a pH of
6.7-7.2, a specific conductance of 70-145 pmhos/cm and a temper-
ature of 25 + 2°C.
Fish, 21-39 millimeters (mm) total length, 0.1-0.4 grams (g)
wet weight were acclimated to 22°C well water over a 24-hour
period. The aforementioned measurements for each group of fish
are based on 30 individuals. The fish were then held at this
temperature for a minimum of 5 days prior to shipment by air to
each site. Upon arrival at a test site, the fish (1,500-5,000)
were transferred to a 600-liter insulated fiberglass holding tank
maintained inside the mobile laboratory. This tank was fitted
with a standpipe drain which maintained a tank volume of approxi-
mately 500 liters. Water flowing into this tank was from the
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potable water supply which had first been dechlorinated (acti-
vated carbon) and adjusted to 22 HH ioc. During the first 18-24
hours of acclimation, the flow rate entering the holding tank
was increased in stages from 0.2 H/minute to 4 £/minute. A flow
rate of 4 2,/minute was then maintained for the duration of the
study and provided 11.5 volume replacements per day.
.Acclimation of fathead minnows to the dilution water (re-
constituted water) was performed twice daily, once in the morning
and once in the afternoon. At each interval, 100-150 fish were
transferred with approximately 15 liters of dechlorinated potable
water to one of two 125-liter stainless steel acclimation tanks.
Reconstituted water .(22°C) was then introduced to each tank at a
flow rate of 250 mi/minute. The tanks were maintained at a con-
stant volume of 90 liters.
The fish were acclimated to reconstituted water for a 24-
hour period prior to use in a test. The holding tank was then
drained and the acclimation process repeated with a new group of
fish.
Fathead minnows in the main holding tank were fed a dry
commercial fish food once a day following the afternoon transfer
of fish to the stainless steel holding tank. Fish were not fed
during the 24-hour acclimation period in the dilution water nor
during the performance of the tests. Cumulative mortality during
the 30-day holding periods prior to shipment of the fish to each
site and during the holding and -acclimation periods on-site did
not exceed 1%.
Water flea—The water flea (Daphnia magna) used in this
study were obtained from laboratory stocks cultured at the
Wareham laboratory. The culture water used for the daphnids was
reconstituted deionized well water characterized by a total hard-
ness of 160-190 mg/A as CaCOs, total alkalinity of 110-130 mg/£
as CaCO3, a pH of 7.8-8.2, a specific conductivity of 500-700
ymhos/cm, a temperature of 21-23°C, and a dissolved oxygen con-
centration greater than 60% of saturation (5.2-5.4 mg/X.) . The
daphnid cultures were fed a diet of unicellular green algae and
commercial fish feed.
Large gravid females were selected from the laboratory cul-
tures and shipped by air in approximately ten liters of culture
water to each testing location. Upon arrival at each test site,
the cultures were transferred to three 60-fc glass aquaria. Re-
constituted dilution water was used to supplement the original'
culture water to yield a total volume of 40 £./aquarium. Water
in each aquarium was then gently aerated. The cultures were
maintained under static conditions at 22 + 1°C. D. magna were
fed daily a commercial fish feed which had" been mTxed with recon-
stituted water in a glass blender cup.
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The daphnids used in the on-site testing were generally
less than twenty-four hours old. The procedure for obtaining the
test organisms was to first transfer an adequate number of gravid
adult daphnids from the cultures to a 2-1 battery jar containing
1.7 St. of reconstituted water. On the following day, the first
instaj? daphnids released from the adults were removed and ran-
domly distributed to the test chambers for testing. On those
occasions when the isolated adults failed to release a sufficient
number of young, the amount required for testing was supplemented
by collecting first instar daphnids from the main culture aquaria.
On-Site Test Procedures
General— Twenty-four hour static acute toxicity tests were
performed daily on samples of "untreated" influent and "treated"
effluent at each of the study sites (Mobay Chemical, only
"treated" effluent) over ten consecutive test days. Fathead
minnow tests were conducted on wastewater samples collected at
two sampling periods each day while daphnid tests were performed
on alternating days beginning with test day 1. At 4 of the sites,
Salmonella Bacterial Mutagenesis Assays (Ames Test) were per-
formed on samples of the "treated" effluent.
Grab samples of each wastewater (influent and effluent) were
obtained twice a day (9:00 a.m./3:00 p.m.) for ten consecutive
days to coincide with the test regimentation. A sample of each
wastewater ^as taken prior to the first day of .testing in order
that preliminary, raage-findiag -tests could -be conducted with
each test species. At certain sites, personnel from the "host"
company collected duplicate or "split" samples of the wastewater
with the laboratory personnel from EG&G, Bionomics,
Each grab sample was collected in a separate polyethylene
bucket at each sampling location and transferred to individual
53-1 NalgeneR polyethylene carboys. The total sample volume
ranged from 20-50 H and was dependent on the toxicity of each
wastewater. Upon delivery to the mobile laboratory facility, a
500 m£ subsample was drawn from each wastewater prior to temper-
ature modification in order to conduct limited wastewater char-
acterization. The wastewater samples were then heated or chilled
as necessary to 22 + 2°C. This was accomplished by pumping each
wastewater sample through individual stainless steel coils which
were immersed in the main diluent tank. Once the desired test
temperature (22°C) of each wastewater was attained, the samples
were immediately used to initiate the respective toxicity tests
scheduled for that day.
A characterization of the chemical and physical parameters
of the dilution water and "treated" and "untreated" effluent
wastewaters was performed daily at each site. On each test day,
samples of reconstituted dilution water were analyzed for total
hardness, total alkalinity, specific conductance, pH, temperature
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and total residual chlorine. Similarly, aliquots of each waste-
water sample collected during each ten day study were analyzed
for dissolved oxygen concentration (DO), pH, specific conductance,
temperature and total residual chlorine. These measurements were
made on samples of the wastewaters brought back to the mobile
laboratory, but prior to temperature adjustment.
During each test, the pH, dissolved oxygen concentration
and temperature were measured in the control, high, middle and
low test concentrations prior to the addition of the -test organ-
isms and at the termination of the test. Specific conductance
-was jnaanirfcored similarly tost only at t&e initiation of "testing.
Total hardness and alkalinity measurements were performed
according to Standard Methods (APHA et al., 1975, pg. 179 and 53,
respectively)"! Specific conductance was measured with a Model
#33 YSI conductivity meter, pH with a Model #175 Instrumentation
Laboratory pH meter and combination electrode and dissolved
oxygen with a YSI Model #57 DO meter. Total residual chlorine
measurements were performed on a Fisher and Porter Model #17T1010
amperometric titrator according to either the "forward" titration
procedure (APHA et al., 1975, p. 112) or the "backward" titration
method (Fisher and Porter Bulletin 17T1010, pg. 7). Temperature
maintained at 22°C was monitored with either a hand held mercury
column thermometer and/or YSI Model #80A recorder. All measure-
ments were performed using the appropriate metering devices pre-
viously described. Each metering device was calibrated once a
day for -ten consecutive test
-------
After twenty-four hours of exposure, observations of mor-
tality and physical/behavioral abnormalities were made and re-
corded for fish in each test aquarium. The water quality para-
meters (pH, DO, temperature) were measured and recorded prior to
test termination. The test aquaria for each wastewater were then
immediately drained, brushed down, and rinsed thoroughly in order
that a new test could be set up immediately in the same respec-
tive aquaria.
Daphnia magna Tests
Daphala magna acutes with Bach wastewater were initiated
using a minimum of five test concentrations plus a control in
triplicate. The dilution factor for each wastewater concentra-
tion was also 0.65.
The diluent water used in these studies was the same as that
described previously. Each test concentration was prepared by
measuring out the wastewater in a Type A 500-mJl graduated cylin-
der. Dilution water was then transferred to a l-£ beaker, mixed
on a magnetic stirrer and quantitatively divided among 3 repli-
cate 250-m£ beakers. The A replicate of each concentration re-
ceived 200 mSt, of test solution while the remaining two replicates
received 150 mZ each. The additional volume in the A replicate
was necessary to accomodate the meter probes used for daily water
chemistry measurements.
As was done in the fish acutes* water quality parameters
were monitored at the beginning of the study prior to addition
of the test organisms and again at the end of the test. Five
daphnids were randomly assigned to each test vessel within 30
minutes after the introduction of each wastewater sample, a total
of 15 daphnids per concentration. All tests were carried out at
22 + 2°C.
Ames Test
Salmonella Mutagenicity Tests (Ames Test) were carried out
on samples of "treated" wastewater from the four industrial
wastewater treatment plants indicated in Table 1. The tests were
conducted at EG&G, Mason Research Institute, Rockville, Maryland.
The specific protocol followed is presented in the Appendix. At
each participating test site, a 100-mJl subsample of the 9:00 a.m.
"treated" wastewater (effluent) sample was collected in a clean
235-m£ glass bottle, sealed with a TeflonR-lined cap and frozen.
Sampling occurred on the first day of testing and then on an
alternate day basis. At the end of the ten consecutive days of
testing, the five samples were packed with dry ice and shipped,
by air, to the Mason Research Institute, where the samples were
thawed out and diluted to the appropriate volume prior to test-
ing.
-------
Statistics
Mortality data derived from each definitive test were used
to calculate a median lethal concentration (LC50) and its 95%
confidence limit utilizing the moving average angle method
(Harris, 1959). The data are presented as both LC50 values
(percent wastewater) and lethal units (100/LC50). The LC50 is
the calculated nominal concentration of the wastewater in diluent
water which produces mortality of 50% of the test animal popula-
tion at the stated time of exposure; i.e., 24-hour LC50. In
those instances where T*C50 values could not be calculated due to
<50% mortality in 100% wastewater, the lethal units have been
derived through graphical interpolation of the data on log x
probit paper and provide the most probable toxicity concentration
(Esvalt, et al., 1971). All data are presented after rounding to
two signifTcant figures.
-------
SECTION 3
RESULTS
Union Carbide, South Charleston, Wesl: Virginia
On-site toxicity tests with fathead minnow and Daphnia magna
were conducted at the South Charleston Waste Treatment Works from
September 27 - October 15, 1978. The wastewater treatment plant
treats both industrial and domestic wastewaters which are dis-
charged to the Kanawha River through a common.discharge pipe.
The industrial portion of the wastewater is derived from the
Union Carbide complex in South Charleston.
Due to its nature, the industrial wastewater stream enters
the plant as a separate influent stream and remains segregated
from the domestic wastewaters throughout the treatment process.
Overflow from the secondary clarifiers of the industrial treat-
ment system combines with the municipal effluent to form the
primary discharge to the river. A flow diagram of the industrial
wastewater treatment system with identification of the sample
points is presented in Figure 1.
Tests with "untreated" influent and "treated" effluent were
performed according to the scope of work described previously.
Results of the acute toxicity studies are presented in Table 2.
These data indicate that the "untreated" influent was relatively
toxic to both test species while no mortality was observed in
either test population exposed to the "treated" effluent through-
out the ten day testing period. The wastewater treatment process
provided a 100% reduction in lethal units (10 units) for fathead
minnow and a 100% reduction (12 units) for daphnids.
Measured water quality parameters of the major test waters,
i.e., influent wastewater (untreated), effluent (treated) and
diluent water (reconstituted) are presented in Table 3. The
data, compiled during the ten consecutive days of on-site testing
are tabularized as the means and range for each parameter. As
would be expected, the measured parameters for the "untreated"
influent varied over a greater range than were observed for the
"treated" effluent.
A summary of the water quality parameters measured in expo-
sure aquaria during the fish and daphnid toxicity tests with each
wastewater are presented in Tables 4 and 5. It should be noted
10
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-------
that the high, middle and low test concentrations for the "un-
treated" influent were changed during the ten days of testing as
a result of the observed toxicity of this wastewater which guided
the. selection of wastewater concentrations to be tested. The
concentration range used for any given test was based on the test
results from toxicity data generated from the previous day. An
attempt was made to try and obtain an effect/no effect level for
each test. The data presented in Tables 4 and 5 indicate that
the dissolved oxygen concentration and pH varied minimally be-
tween treated effluent concentrations for either species. A
wider range of values were observed during the testing of the
"untreated" influent. These data also indicate that the specific
conductance varied directly with the influent/effluent concentra-
tion.
Samples for the Salmonella mutagenicity tests (Ames test)
were not obtained at this site.
Monsanto, Muscatine, Iowa
On-site toxicity tests with fathead minnow and Daphnia magna
were conducted at the Monsanto Company, Muscatine, Iowa from
October 17-29, 1978. The wastewater treatment facility consists
of two separate treatment plants. One plant (ABS Plastics)
handles wastewaters from the production of intermediate chemicals
for the production of plastics, while the second plant treats
wastewaters from the agricultural chemicals facility. Treated
wastewaters from the treatment plants combine with large amounts
of non-contact cooling water from -the 001 discharge entering the
Mississippi River. For the purpose of this contract, only the
treated effluent from the Agricultural Chemicals treatment plant
was tested. "Untreated" influent was obtained prior to pH adjust-
ment while "treated" effluent was obtained from the overflow of
the primary clarifiers. A flow diagram of the wastewater treat-
ment process is presented in Figure 2.
Results of the acute toxicity studies with the "untreated"
influent and "treated" effluent are presented in Table 6. As
expected, the "untreated" influent was quite toxic to both test
species. The mean number of lethal units (LCSO's) contained in
the influent were 47(2.3%) and 41(2.9%), respectively, for
fathead minnows and daphnids. Results of the "treated" effluent
studies indicate a 97% reduction in toxicity after treatment for
both fathead minnows (1.5 lethal units, 68%) and water flea (1.3
lethal units, 75%).
The measured water quality parameters of the "untreated"
influent, "treated" effluent and reconstituted diluent water com-
piled during the ten consecutive days of on-site testing are pre-
sented in Table 7. The pH and conductivity of the influent ex-
hibited considerable variation from one sample to the next, while
for the "treated" effluent all of the measured parameters remained
12
-------
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FIGURE 2. FLOW DIAGRAM FOR WASTE TREATMENT FACILITY AT MONSANTO, MUSCATINE, IOWA.
-------
relatively constant.
A summary of the water quality parameters measured in the
exposure aquaria during the fathead minnow and daphnid toxicity
tests with each wastewater are presented in Tables 8 and 9,
Variations in the pH and specific conductance of the "untreated"
influent were directly correlated with the toxicity of the waste-
water. Consequently, the data presented for the high, middle
and low test concentrations represent ranges for the respective
doSe' levels which varied accordingly from day to day.
The water quality parameters monitored during the tests with
both wastewaters remained relatively constant within the same set
of concentrations. This probably is a result of the moderating
effects of the dilution water which was used in proportionally
greater volumes in setting up the "untreated" influent tests.
Even at these low test concentrations a significant oxygen demand
was present and may have contributed to the observed mortalities.
The results of the Salmonella mutagenicity assay of the
"treated" effluent sampled on days 1, 3, 5, 7 and 9 indicate
that there was no consistent significant increase in the rever-
sion index of any of the tester strains with or without metabolic
activation.
Mobay Chemical, Kansas City, Missouri
On-site toxicity tests were conducted at the Mobay Chemical
Company, Kansas City, Missouri during the period November 3-7,
1978. On November 2, EG&G, Bionomics was informed that the
Environmental Protection Agency had accepted Mobay's request to
omit the tests with the "untreated" influent and consequently
the on-site tests were limited only to the "treated" effluent.
The "treated" effluent was obtained from the biological effluent
flume; a concrete raceway where the outfall from two clarifiers
are channeled. After leaving the flume, these wastewaters are
combined with the discharge from the Kansas City Electric Gener-
ating Plant, which subsequently flows into the Blue River. A
flow diagram of the wastewater treatment plant at Mobay was not
provided.
Results of the acute toxicity studies with the ."treated"
effluent are presented in Table 10. These data indicate that
the mean acute LC50 value for fathead minnow and daphnids exposed
to the "treated" effluent were 28% (3.8 lethal units) and 23%
(4.4 lethal units), respectively. The efficiency (in terms of
% reduction of lethal units) of the wastewater treatment facility
could not be determined as a result of the omission of the tests
with "untreated" influent.
The measured water quality parameters of the "treated"
effluent and reconstituted diluent water, monitored during the
14
-------
on-site testing, are presented in Table 11. These data suggest
that no significant variation was observed throughout the testing
.period..
A summary of water quality parameters measured in the high,
middle and low exposure concentrations and control during the
fathead minnow and daphnid toxicity tests with the "treated"
effluent are presented in Table 12. These data indicate little
variation in the initial dissolved oxygen concentration or pH
for each .test throughout the .testing period,. Specific conduct-
ance varied directly with the concentration of effluent. Dis-
solved oxygen concentrations observed at the termination of the
fathead minnow tests, though significantly reduced from the 0-
hour reading, were generally adequate to support aquatic life.
The results of the Ames Test performed on the "treated"
effluent water samples from Mobay Chemical, obtained on test day
1, 3, 5, 7 and 9 indicate that none of the samples caused a sig-
nificant increase in the reversion index of any of the test
strains with or without metabolic activation.
Monsanto, Luling, Louisiana
On-site toxicity tests with fathead minnow and Daphnia magna
)apf
were conducted at Monsanto, Luling, Louisiana during the period
December 1-17, 1978. The Luling complex manufactures a number of
chemical products in addition to the glyphosate product RoundupR.
The wastewater treatment plant received the wastewater from the
Roundup facility as well as from the other manufacturing pro-
cesses. Therefore, the samples of "untreated" influent and
"treated" effluent were representative of the combined processes
of the Luling complex. Samples of the influent were obtained
from the overflow of the collection basin after acid-base neutra-
lization, while the "treated" effluent was obtained from the dis-
charge weir of the aeration lagoon. A flow diagram of the waste-
water treatment system is presented in Figure 3.
s,
Table 13 presents the results of the 24-hour static acute
toxicity tests with "untreated" influent and "treated" effluent.
The "untreated" influent exhibited a mean of 74 lethal units
(LC50, 1.5%) for fathead minnows and 88 lethal units (LC50, 1.3%)
for the water flea. After treatment, the mean number of lethal
units for fatheads and daphnids were 6 (LC50, 18%) and 3 (LC50,
33%), respectively. Although these values indicate that the
"treated" effluent is still moderately toxic to both test species
they do represent 92% (fathead minnow) and 96% (daphnid) reduc-
tion in toxic units by the wastewater treatment facility.
Water quality parameters monitored on the influent, effluent
and dilution water during the testing period are presented in
Table 14. A summary of the water quality parameters measured in
the exposure aquaria during the fathead minnow and daphnid toxic-
15
-------
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FIGURE 3. FLOW DIAGRAM FOR WASTE TREATMENT FACILITY AT MONSANTO, LULING, LOUISIANA.
-------
ity tests with each wastewater is presented in Tables 15 and 16.
The high, middle and low test concentrations for each wastewater
were varied on a day to day basis, when necessary, in an attempt
to obtain effect/no effect tests. The data presented in Tables
15 and 16 indicate that the initial (0-hour) dissolved oxygen
concentration and pH varied minimally in both wastewaters and
that the specific conductance varied directly with the influent/
effluent concentration. The measurements made at the end of the
fathead minnow tests with "treated" effluent .indicate .DO concen-
trations in the high concentration were at times marginal.
The results of the Ames Test performed on the "treated"
effluent water samples collected on test day 1, 3, 6, 7 and 9
indicate a weak but consistent increase in the reversion index
of tester strain TA 1535. The dose related increase was present
both with and without metabolic activation but was most pronoun-
ced in the presence of rat liver microsomes.
A confirmation study of the His"1" phenotype of "revertant"
colonies of strain TA1535 at the highest dose level indicated
that 91% of the colonies were true revertahts. This result
supports the conclusion that the observed increase in reversion
index of TA1535 is real and not an artifact of the experiment.
Diamond-Shamrock, Green Bayou, Texas
On-site toxicity tests with fathead minnow and Daphnia magna
were conducted at Diamond-Shamrock's Green Bayou, Texas plant
during the period January 3-22, 1979. The Green Bayou complex
manufactures a number of chemical products in addition to its
major product Dakonyl^, a fungicide. Samples of the "untreated"
influent were obtained from the inflow pipe prior to introduction
into the primary settling basin. Samples of "treated" effluent
were obtained from the discharge raceway of the aeration lagoon.
A flow diagram of the wastewater treatment facility was not pro-
vided.
Results of the acute toxicity studies with the "untreated"
influent and "treated" effluent are presented in Table 17. The
"untreated" influent was considerably more toxic to fathead
minnow (mean = 360 lethal units, LC50, 3.3%) than to the water
flea (mean = 130 lethal units, LC50, 2.1%). After treatment,
the toxicity of the wastewater was reduced to 1.2 lethal units
(LC50, 84%) for fathead minnows and 2.6 lethal units (LC50, 38%)
for Daphnia magna. This is a reduction in lethal units of 99.7%
and 98%for the fish and daphnids. In comparing the mean LCSO's
to the mean lethal units for the "untreated" influent tests, it
would appear that the values are incorrect in that the mean LC50
divided into 100 does not equal the mean lethal units. Results
of several of the acute tests indicate that at times the "un-
treated" influent was extremely toxic. When these LC50 values
are converted to lethal units, they tend to skew the mean.
17
-------
Water quality parameters monitored for the two wastewaters
and dilution water are presented in Table 18. Table 19 and 20
present a summary of the water quality parameters measured in the
high, middle and low exposure aquaria including the control
during toxicity tests with fish and daphnids. As in previous
tes'ts with other wastewaters, the dissolved oxygen concentrations
and specific conductance varied directly with the wastewater
concentration. DO levels were again marginal in the high concen-
trations of the fish tests with both the influent and effluent.
The results of the Ames Test performed on the "treated"
effluent water samples from the Diamond-Shamrock plant in Green
Bayou, Texas on days 1, 3, 5, 7 and 9 indicate that none of
these samples caused a significant increase in the reversion in-
dex of any of the tester strains with or without metabolic acti-
vation.
E.I. duPont de Nemours, LaPorte, Texas
On-site toxicity tests with fathead minnows and Daphnia
magna were conducted at duPont's LaPorte, Texas plant during the
period February 9-24, 1979. The LaPorte complex manufactures a
number of chemical products and pesticides which includes Maneb
and Methomyl. Samples of two "treated" influents were collected
at this site. One influent wastewater (Stream #1) was collected
from the process wastewater ditch after pH adjustment. This
wastewater stream was comprised of several pesticide/non-pesti-
cide unit effluents and constituted the main flow into the treat-
ment system. The second influent (Stream #2) was collected from
the Maneb/Methomyl wastewater stream prior to chemical oxidation
with chlorine. The "treated" effluent was obtained from a spigot
located at the outfall pond #101. The "treated" effluent com-
bines with the non-contact wastewater from outfall #201 prior to
discharge into San Jacinto Bay. A flow diagram of the wastewater
treatment facility was provided to EG&G, Bionomics labelled con-
fidential and therefore is not included in this report.
Results of the acute toxicity studies with fathead minnow
and daphnids to two "treated" influents and the "treated" effluent
are presented in Tables 21 and 22. A complete regimentation in
testing of the "treated" influent collected from the Maneb/
Methomyl production stream could not be accomplished due to a
malfunction in the plant equipment and subsequent process shut-
down on test day #4. The data from the acute toxicity studies
indicate that the mean number of lethal units (LC50) for fathead
minnow exposed to the "treated" influent (pesticide/non-pesticide
stream #1) was 4.5 lethal units (27%), while the partial testing
of the "treated" influent (Maneb/Methomyl prior to chemical oxi-
dation, stream #2) resulted in an acute toxicity of 140 lethal
units (0.75%). The mean for fathead minnow exposed to the
"treated" effluent was 2.4 lethal units (42%). This indicates
that the wastewater treatment process provided 47% and 95% reduc-
18
-------
tion in toxicity (2.2 and 134.6 lethal units) to the two "treat-
ed" influents, respectively.
The mean number of lethal units (LC50) for daphnids exposed
to the two "treated" influents (pesticide/non-pesticide and
Maneb/Methorny1) and the "treated" effluent were 4.8(28%), 770
(0.14%) and 35(4.4%), respectively. These data would indicate
that the wastewater treatment process provided a 95% reduction
in toxicity to daphnids exposed to the Stream 12 influent (Maneb
Methomyl), but that no reduction in toxicity was observed when
compared to the results of the Stream fl influent. Due to the
physical limitations inside the mobile laboratory, all of the
incoming wastewater streams entering the treatment plant could
not be tested. Therefore, an accurate interpretation of the
efficiency of the duPont wastewater treatment facility cannot be
made.
The measured water quality parameters of the "treated"
influent (Stream #1), the "treated effluent and reconstituted
diluent water, compiled during the ten consecutive days of on-
site testing, are presented in Table 23. These data suggest
that the "treated" influent was more variable with respect to all
measured parameters than was the "treated" effluent.
A summary of the water quality parameters measured in the
exposure aquaria during the fathead minnow and daphnid toxicity
tests with each wastewater are presented in Tables 24-27. These
data indicate that the greatest variation in any measured para-
meter was the pH observations for the "treated" influent from
the pesticide/non-pesticide stream #1. The dissolved oxygen
concentrations observed after 24 hours in all three test waters
decreased in direct proportion to the test concentration while
the specific conductance increased in direct relation to the test
concentrations.
Samples of the Salmonella/mammalian-microsome mutagenicity
test (Ames Test) were omitted at this test site at the request
of the Project Officer.
International Paper, Georgetown, South Carolina
Toxicity tests with fathead minnow and Daphnia magna were
conducted on-site at the International Paper plant in South
Carolina during the period March 19-April 3, 1979. The George-
town complex is considered, in'the paper mill industry, as being
a bleached-kraft, semi-chemical paper mill. The samples of
"untreated" influent were collected from an open canal which
collected the outfall from each mill process, i.e. primary clari-
fier, kraft pulping, bleach plant, cooling tower blowdown, tall
oil, caustic waste and lime mud waste, prior to chemical treat-
ment. The exact size of sample collection was just prior to the
chemical treatment weir. The "treated" wastewater is then pumped
19
-------
to ;a large aeration pond prior to being pumped into a retention/
settling pond. The samples of "treated" effluent were collected
at the pumping gate of the retention pond where the effluent was
discharged into a canal which emptied into the Sampit River and
subsequently Winyah Bay. Discharge of the effluent was conducted
twice a day to correspond with the high ebb in the tidal zone.
A flow diagram of the wastewater treatment process is presented
in Figure 4.
The results of the acute toxicity studies with fathead
minnow and daphnids exposed to the "untreated" influent and
"treated" effluent are presented in Table 28. These data indi-
cate 2.8 and 1.7 lethal units, respectively, were present in the
"untreated" influent as determined from tests with fathead minnow
and Daphnia magna. No mortality was observed in any of the tests
with fathead minnows exposed to the treated effluent. In the 10
tests performed with daphnids and the effluent, none of the test
concentrations produced more than 7% mortality. A 100% reduction
in lethal units was provided for both the fathead minnow and
Daphnia magna by the wastewater treatment process.
The results of the water quality parameters for the dilution
water,influent and effluent are tabularized in Table 29. These
data suggest that minimal variations occurred within each waste-
water.
A summary of the water quality parameters measured in the
exposure aquaria of the high, middle and low test concentrations
including the control during the performance of the tests with
both species are presented in Tables 30 and 31. These data in-
dicate very minimal variations in the pH of each wastewater and
that the depletion of the dissolved oxygen concentration in each
test followed similar patterns observed at other sites and was
directly related to test concentration. Specific conductance
also varied in direct proportion to the test concentration.
Samples for the Ames Test were omitted at this test site at
the request of the Project Officer.
20
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SECTION 4
DISCUSSION
On-site 24-hour static acute toxicity studies were performed
on the wastewaters from 5 pesticide manufacturing plants, an or-
gano-chemical manufacturer and a bleached kraft paper mill plant.
The majority of the wastewater treatment systems evaluated con-
sisted of the following (generalized) treatment steps', coarse
screening of particulates, pH neutralization, activated sludge
digestion and clarification. The findings generated by the
contract will provide useful and relavent information both to
industry and Federal and State regulatory agencies.
In the present study, the results of toxicity studies indi-
cate a higher level of lethal units present in the wastewaters of
the pesticide formulation, both before and after treatment as
compared to the wastewaters of the pulp and paper mill and organo-
chemical plant. A sufficient data base is not available to de-
termine whether these observations are valid overall for the
industries of concern. The average efficiency "of the wastewater
treatment plants in reducinjg--fehe-^oxicity of the "untreated"
wastewater was 98% Tralig^^^2-10Q%j>, Excluded in the average
were Mobay Chemical Company^aTrdE. I. duPont. Mobay was excluded
since influent tests were not performed and consequently the
percent efficiency of the treatment system could not be deter-
mined. E.I. duPont was excluded from the calculation since a
true representation of the toxicity of the entire influent waste-
water load could not be determined.
The results of ten consecutive days of biological testing
at each site indicate that the toxicity of the "treated" effluent
samples remained relatively constant from day to day. The aver-
age number of lethal units present in the "treated" effluents,
as determined from tests with fathead minnows, was 2.1 with a
range of 0-6 lethal units. Similarly, Daphnia magna tests pro-
vide a mean of 6.5 lethal units with a range of 0-34. As would
be expected the toxicity of the "untreated" influent samples was
extremely variable on a day to day basis. From the fathead
minnow studies, the "untreated" influent contained a mean of 93
lethal units (range, 4.6-373), while Daphnia magna acute tests
produced a mean of 261 lethal units (range, 1.7-782).
It is interesting to note that the toxicity of the "treated"
22
-------
effluent is relative to the toxicity of the "untreated" influent.
For example, if an influent contained 300 lethal units and under-
went biological and/or chemical treatment which has an efficiency
of 99%, then the "treated" effluent would still contain 3 lethal
units. This would be equal to an LC50 value of 33% effluent.
Conversely, if an influent having moderate toxicity is subjected
to the same treatment efficiency, the resulting effluent will
probably be non-toxic or of a very low toxicity. The signifi-
cance of this observation must be viewed in terms of what addi-
tional engineering steps have been applied to the effluent be-
tween the time the wastewater leaves the treatment system and
enters the receiving water. Dilution with other treated process
streams and non-contact cooling water must be taken into consid-
eration in both designing wastewater treatment system and regu-
lating their discharge.
The Mass Emission Rate as proposed by Esvelt et al. provides
a useful method of incorporating lethal units and flow rates in
assessing the impact of a wastewater stream on the total ecology
of the receiving water. To determine the mass emission rate for
a stream, one must first determine the relative toxicity (RT) of
the wastestream which is defined as the product of the toxicity
concentration (expressed as lethal units) and the flow rate.
RT = Tc X Flow Rate
The mass emission rate then for a receiving water (TC2) is
defined as the relative .toxicity divided by the combined waste
and dilution water flow.
TC2 = RT
wastewater . dilution water
flow rate flow rate
At present, the mass emission rate for a receiving water is
a somewhat nebulous value requiring further regulatory defini-
tion. If a program for water quality management is to be devel-
oped, which considers the toxic properties of wastes, then addi-
tional information must be obtained on the efficiency of the
wastewater treatment systems. The utilization of a study program
similar to that carried out under the present contract is one
method by which the minimum efficiency of a wastewater treatment
plant, under normal operating conditions, could be determined.
If the toxicity of an effluent could be correlated with one or
more chemical parameters which can be monitored continuously,
then a mechanism would evolve by which regulators and plant
operators could monitor the discharge of toxic wastes on a real
time basis before they had a chance to cause damage to the envir-
onment .
Of the two species tested, neither the fathead minnow nor
the water flea proved to be routinely more sensitive than the
other in tests with the "untreated" or "treated wastewaters.
23
-------
J 3
Since an adequate data base has not been developed for these
species to the vast majority of industrial effluents, it is sug-
gested that both the fathead minnow and daphnids be tested. On
the other hand, should an on-going biomonitoring program be
instituted for an industrial discharge in which both species
provide similar results, then future testing might be limited to
only one species. Daphnia magna are attractive for this purpose
from the standpoint that their use can greatly reduce the eco-
nomic burden to an industry for such a program. Daphnids are
relatively easy to culture, require no capital investment and can
be used by plant personnel to generate required data. It may
also be assumed that with such a biomonitoring tool available to
them, industry will take it upon themselves to incorporate its
use on a regular basis in monitoring the efficiency of their
wastewater treatment systems.
Finally, the use of the lethal unit concept should be appli-
ed to all effluent toxicity studies. LC50 values as percent
effluent do not provide an accurate assessment of the impact of
the wastewater. This is especially true for highly toxic wastes.
The mean of a number of LC50 tests may not equal the mean from
the same tests expressed as lethal units. A highly toxic effluent
when converted to toxic units will skew the mean upwards and as a
result will more accurately represent the toxic load entering the
environment.
For those effluents resulting in less than 50% mortality in
100% effluent, the most probable toxicity concentration (expres<-
sed as lethal units) is equally important in determining mass
emission rates.
24
-------
SECTION 5
REFERENCES
APHA, AWWA, WPCF. 1971. Standard methods for the examination
of water and wastewater. 13th Edition, Washington, D.C.
1193 pp.
Esvelt, L. A., W. J. Kaufman and R. E. Selleck. October, 1971.
A study of toxicity and biostimulation in San Francisco
Bay - Delta Waters. Vol. IV. Sanitary Engineering Research
Laboratory. Univ. California, Berkeley.
Harris, E. K. 1959. Confidence limits for the LD50 using the
moving average-angle method. Biometrics, Vol. 4, #3. pp.
157-164.
U.S. EPA. 1975. Methods for acute toxicity tests with fish,
macroinvertebrates and amphibians. Ecological Research
Series (EPA-660/3-75-009), 61 pp.
25
-------
SECTION 6
TABLES
TABLE 1. LIST OF THE SEVEN INDUSTRIAL WASTEWATER TREATMENT
FACILITIES WHICH WERE CHOSEN FOR ON-SITE EFFLUENT
TOXICITY STUDIES.
Mpdif ied_ list.
of sites
. Dates, of _pn-si_te.
testing
Ames_
test
Union Carbide,
S. Charleston, W. Virginia
Monsanto,
Muscatine, Iowa
Mobay Chemical,
Kansas City, Missouri
Monsanto,
Luling, Louisiana
Diamond-Shamrock,
Green Bayou, Texas
duPont,
LaPorte, Texas
International Paper,
Georgetown, So Carolina
9/27/78-10/15/78
10/17/78-11/2/78
11/3/78-11/17/78
12/1/78-12/17/78
1/3/79-1/22/79
2/8/79-2/27/79
3/20/79-4/2/79
no
yes
yes
yes
yes
no
no
26
-------
V
TABLE 2. LCSO VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS AND DAPHNIA MAGMA EXPOSED TO "UNTREATED INFLUENT" AND "TREATED EFFLUENT" FROM THE SOUTH
CHARLESTON, WEST VIRGINIA INDUSTRIAL HASTEWATER TREATMENT PLANT.
Day
Fathead Minno
24-hour LCSO
(» influent)
lethal units
/ »
1
w - Untrcate
IS
6.7
9.5
10
2
d Infl
23
4.3
uent
18
5.6
Fathead Minnow - Treated Effluent
24-hour LCSO
(» effluent)
lethal units
>100
0
MOO
0
MOO
0
MOO
0
•Daphnia magna - Untreated Influent
24-hour LCSO
(\ influent)
•lethal units
21
4.8
17
5.9
•
_
Daphnia magna - Treated Effluent
24-hour LCSO
(» effluent)
lethal units
MOO
0
>100
0
-
-
3
15
6.7
MOO
0
26
3.8
MOO
0
12
a. 3
MOO
0
21
4.8 •
MOO
0
4
25
4.0
MOO
0
-
-
15
6.7
MOO
0
-
-
5
28
3.6 .
MOO
0 •
19 ..:
5L3 ..
MOO
0.
18
3.6
MOO
0
14
7.1
MOO
0
6
22
4.5
MOO
0
-
-
13
7.7
MOO
O
-
-
7
2.0
50 -
MOO
0
1.8
56
MOO
0
L6
6.3
MOO
0
13
7.7
MOO
0
a
7.5
13
MOO
0
-
-
12
8.3
MOO
0
-
-
9
22
4.5
MOO
0
SO
2.0
MOO
0
2.6
38
MOO
0
4.7
21
MOO
0
10
28
3.6
MOO
0
-
-
18
S.6
MOO
0
-
-
Mean
16
10
MOO
0
19
12
MOO
0
St.
Dev.
7
.12
13
16
-
Tests not required at this time period.
-------
TABLE 3. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURING TEW-
CONSECUTIVE DAYS OF ON-SITE TESTING AT THE SOUTH CHARLESTON, WEST VIRGINIA
INDUSTRIAL WASTEWATER TREATMENT PLANT
00
Temperature (°C)
PH
Dissolved oxygen
(mg/i,)
Conductivity
(ymhos/cm)
Hardness (mg/£)
Alkalinity (mg/£)
Chlorine (mg/£)
Diluent
Mean
22
8.3
*
NA
470
170
110
NDf
water
Range
21-23
7.9-8.4
NA
440-500
160-170
110-120
ND
Treated
Mean
22
7.6
6.2
1800
NA
NA
ND
effluent
Range
18-24
7.2-7.8
4.9-7.6
1400-2300
NA
NA
ND
Untreated
Mean
27
9.6
7.0
2600
NA
NA
ND
effluent
Range
24-31
1.8-12
3.6-8.2
700-7000
NA
NA
ND
Analyses not routinely performed.
1' •
ND = non-detectable, e.g. below detectable limit of 0.01 mg/fc,
-------
TABLE 4. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOWS TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE SOUTH CHARLESTON, WEST VIRGINIA INDUSTRIAL
WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE OF
MEASUREMENTS OBSERVED DURING THE TEN CONSECUTIVE DAYS OF ON-SITE
TESTING.
Concen-
tration
Influent
control
42-7. 5%f
18-3:2%f
7.5-1.3%f
Effluent
control
100%
42%
18%
Dissolved oxygen
(mg/i)
Hour
0
24
0
24
0
24
0
24
0
24
0
24
0
24
0
24
A.M.
7.9-9
6.8-7
8.1-9
0.4-7
8.1-9
0.3-6
7.9-8
0.4-6
8.1-9
6.2-8
5.9-7
3.8-6
7.4-8
5.2-7
8.0-8
6.0-7
.1
.9
.2
.7
.0
.0
.9
.6
.0
.2
.9
.8
.4.
.5
.8
.7
P.M.
7.8-9
6.8-8
6.5-8
0.2-7
7.7-8
0.3-6
7.8-8
0.5-7
7.8-9
7.0-8
5.1-7
3.9-6
7.0-8
5.7-7
7.6-8
6.7-7
.1
.0
.7
.3
.8
.6
.8
.6
.0
.1
.6
.7
.4
.3
.9
.8
8
8
3
4
5
6
6
6
8
8
7
7
7
7
7
7
PH
A.M.
.2-8.4
.0-8.2
.8-11
.0-10
.8-10
.6-8.6
. 8-9 . 6
.9-9.2
.2-8.4
.0-8.2
.2-7.8
.4-8.0
.5-7.9
.6-8.0
.8-8.1
.8-8.1
Conductivity
(umhos/cm)
8
8
9
8
8
8
8
8
8
8
7
7
7
7
7
7
P.M.
.2-8.4
.0-8.2
.1-11
.8-10
.9-10
.8-9.5
.5-9.5
.0-9.1
.1-8.4
.0-8.2
.2-7.8
.4-8.0
.5-8.0
.6-8.0
.8-8.3
.9-8.1
A.M.
440-470
NA*
500-3100
-•NA---"
470-1700
NA
450-1000
NA
440-490
NA
1400-2400
NA
850-1200
NA
650-800
NA
P.M.
420-480
NA
440-1100
NA
480-730
NA
460-600
NA
440-480
NA
1400-2300
NA
850-1200
NA
600-800
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
29
-------
TABLE 5. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF DAPHNIA MAGNA TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE SOUTH CHARLESTON, WEST VIRGINIA INDUSTRIAL
WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE
] OF MEASUREMENTS OBSERVED DURING THE FIVE DAYS OF TESTING.
Concen-
tration
Influent
control
50-5. 8f
21-2. 5%f
9.0-1.0%f
Effluent
control
100%
42%
18%
Dissolved oxygen
(mg/Jl)
Hour
0
24
0
24
0
24
0
24
0
24
0
24
0
24
0
24
A.M.
7.3-8.5
7.3-8.2
6.5-8.6
5.7-8.2
7.1-8.6
3.4-6.4
7.2-8.8
2.1-7.3
7.6-8.2
7.3-8.4
5.9-7.2
6.1-7.9
7.1-7.9
7.0-8.3
7.4-8.1
7.0-8.4
P.M.
7.3-8.4
7.2-8.2
7.1-8.4
2.5-7.8
7.2-8.4
0.3-6.2
7.1-8.5
1.6-7.1
7.1-8.4
7.0-8.2
4.9-7.3
6.0-7.4
6.6-8.0
6.8-7.8
7.0-8.3
7.0-8.0
pH
A.M.
8.2-8.4
8.2-8.3
4.7-10
4.8-9.3
6.4-10
7.2-9.2
7.0-9.7
7.6-8.9
8.2-8.3
8.2-8.3
7.2-7.8
7.9-8.4
7.6-8.0
8.1-8.4
8.0-8.1
8 . 2-8 . 3
P.M.
8.2-8.3
8.0-8.3
10-11
8.9-9.4
9.2-10
8.3-9.0
9.0-9.6
8.1-8.8
8.2-8.3
8.1-8.3
7.2-7.7
7.8-8.4
7.6-7.9
8.0-8.4
7.9-8.1
8.1-8.3
Conductivity
(umhos/cm)
A.M.
430-470
NA
500-3200
NA
495-1700
NA
470-800
NA
420-460
NA
1400-2000
NA
850-1100
NA
610-700
NA
P.M.
430-460
NA
600-1100
NA
510-750
NA
470-600
NA
430-470
NA
1400-2100
NA
860-1200
NA
610-750
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
_ 30
-------
TABLE 6. LC50 VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS fliffi OAPHNIA MAGNA EXPOSED TO "UNTREATED INFLUENT" AND "TREATED EFFLUENT" FROM THE
MONSANTO, MUSCATINE, IOWA INDUSTRIAL HASTEWATER TREATMENT PLANT.
Day
Fathead Minno
24-hour LCSO
(% influent)
lethal units
/ 1
aTmTl
p.m.
** - Untreate
<1.8
>56
4.2
24
2
a.m.
d Infl
2.2
45
p.m.
uent
<1.8
56
Fathead Minnow - Treated Effluent
24-hour LC50
(» effluent)
61
81
56
65
Daphnia magna - Untreated Influent
24-hour LCSO
(» influent)
lethal units
<1.8
>56
2.7
37
*
-
Daphnia maqna - Treated Effluent
24-hour LC50
(\ effluent)
lethal units
65
1.5
B8
1.1
—
-
3
a.m.
3.4
29
65
15 .
>1OO
0
p.m.
1.4
71
Bl
59
85
1.2
4
a.m.
2.1
48
81
-
-
P.O.
2.2
43
81
- '
-
5
a.m.
1.5
67
81
32
78
1-.3
p.m.
2.2
45
52
50
78
1.3
6
a.m.
1.8
56
52
-
-
p.m.
4.2
24
52
-
-
7
a.m.
<.OS
200
52
43
64
1.6
p.m.
2.2
45
.65
37
73
1.4
8
a.m.
1.5
67
65
-
-
p.m.
2.2
45
81
-
-
9
a.m.
2.2
45
65
29
65
1.5
p.m.
2.2
45
65
71
81
1.2
10
a.m.
1.5
67
65
-
-
p.m.
2.7
37
81
-
-
Mean
2.3
47
68
2.9
41
75
1.3
St.
Dev.
0.85
14
12
1-5
17
9.0
0.2
Testa not required at this time period.
-------
TABLE 7. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURINJ? TEN
CONSECUTIVE DAYS OF ON-SlTfi TESTING AT THE MONSANTO, MUSCATINE, IOWA INDUS-
TRIAL WASTEWATER TREATMENT
to
Temperature (°C)
PH
Dissolved oxygen
(mg/£)
Conductivity
(pmhos/cm)
Hardness (mg/£)
Alkalinity (mg/£)
Chlorine (mg/£)
Diluent
Mean
23
8.4
NA
560
170
120
-f 0
watef
Range
22-27
8.3-8,4
NA
480-750
160-190
120-130
.01-0.03
Treated
Mean
27
7.3
6.6
5200
NA
NA
ND1?
effluent
Range
26-29
7.2-7.4
6.0-7.6
4700-5800
NA
NA
ND
Untreated
Mean
30
5.6
7.2
5000
NA
NA
_t
effluent
Range
24-34
4.6-9.1
6.6-8.1
2600-12000
NA
NA
0.03-10
Analyses not routinely performed.
h
Only 2 positive Cl2 readings out of 20 readings.
ND - non-detectable, e.g. below detectable limit of 0.01 mg/H,.
n
-------
TABLE 8. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOWS TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE MONSANTO, MUSCATINE, IOWA INDUSTRIAL WASTEWATER
TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE OF MEASUREMENTS
OBSERVED DURING THE TEN CONSECUTIVE DAYS OF TESTING.
Concen-
tration
Influent
control
10-6. 5%f
4.2-2.7%f
1.8-0.49%'
Effluent
control
100%
42%
18%
Dissolved oxygen
(rag/t)
Hour
0
24
0
24
0
24
*" 0
24
0
24
0
24
0
24
0
24
A.M.
8.6-9.0
7.2-8.2
8.6-9.2
0.2-8.4
8.6-9.2
0.2-6.4
8.4-9.1
2.5-6.4
8.7-9.1
7.4-8.4
6.6-7.6
5.7-7.4
8.0-8.6
6.6-7.7.
8.6-8.9
7.2-8.1
P.M;
8.6-9.1
7.7-8.5
8.7-9.2
0.2-8.7
8.7-9.2
0.4-8.6
8.6-9.1
4.6-8.4
8.6-9.0
7.8-8.4
6.4-7.4
6.1-8.3
8.0-8.6
7.1-7.9
8.5-9.0
7.6-8.2
PH
A.M.
8.2-8.4
8.2-8.3
6.5-8.8
6.8-8.7
7.3-8.7
7.2-8.4
8.0-8.5
7.5-8.1
8.2-8.4
8.2-8.3
7.3-7.4
7.6-7.7
7.5-7.6
7.7-7.8
7.8-7.9
7.9-8.0
P.M.
8.2-8.4
8.2-8.4
7.2-7.9
7.0-7.9
7.6-8.2
7.2-8.0
7.9-8.3
7.5-8.2
8.2-8.4
8.2-8.4
7.2-7.8
7.1-7.8
7.5-8.2
7.8-7.9
7.8-8.4
8.0-8.1
Conductivity
(umhos/cm)
A.M.
470-680
NA
620-1600
NA
540-760
NA
500-600
NA
480-720
NA
4400-5200
NA
2000-2400
NA
1300-1500
NA
P.M.
480-650
NA
640-950
NA
570-710
NA
520-690
NA
470-650
NA
4400-5300
NA
2300-2500
. NA
1300-1400
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on toxicity
results from the previous day.
33
-------
TABLE 9. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF DAPHNIA MAGNA TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE MONSANTO, MUSCATINE, IOWA INDUSTRIAL WASTE-
WATER TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE OF
MEASUREMENTS OBSERVED DURING THE FIVE DAYS OF TESTING.
Concen-
tration Hour
Influent
control
10-4. 2%f
4.2-1.8%t
l.B-0.75%1
Effluent
control
100%
42%
18%
0
24
0
24
0
24
h 0
24
0
24
0
24
0
24
0
24
Dissolved oxygen
(mg/fc)
A.M.
8.6-8.8
8.0-8.4
8.6-8.9
1.7-8.2
8.6-8.9
4.8-8.0
8.6-8.9
5.4-7.6
8.6-8.8
8.0-8.4
6.7-7.8
7.6-8.0
7.8-8.3
8.0-8.3
8.1-8.6
8.2-8.5
P.M.
8.6-8.9
8.2-8.8
8.5-9.1
2.5-6.4
8.6-9.1
5.1-6.8
8.2-9.0
6.8-7.8
8.6-8.9
8.1-8.7
6.7-7.5
7.4-8.2
8.0-8.4
7.9-8.5
8.4-8.8
8.0-8.8
PH
A.M.
8.3-8.4
8.2-8.4
7.5-8.8
7.4-8.6
7.8-8.7
7.6-8.4
8.1-8.5
7.5-8.4
8.3-8.4
8.2-8.4
7.4
8.0-8.1
7.6
8.1-8.2
'7.8-8.1
8.2-8.3
P.M.
8.4
8.2-8.4
7.6-8.2
7.4-8.0
8.0-8.3
7.7-8.1
8.0-8.3
7.8-8.3
8.3-8.4
8.2-8.4
7.2
8.0-8.1
7.6-7.7
8.1-8.2
7.8-8.0
8.2
Conductivity
(ymhos/cm)
A.M.
480-670
6
NA
650-980
NA
580-750
NA
540-680
NA
480-670
NA
4300-5100
NA
2200-2500
NA
1200-1300
NA
P.M.
480-650
NA
550-780
NA
530-720
NA
510-680
NA
460-640
NA
4400-4800
NA
2200-2500
NA
1200-1300
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
34
-------
TABLE 10. LC50 VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS AMD DAPHNIA MAGHA EXPOSED TO "TREATED EFFLUENT" FROM THE MOBAY CHEMICAL, KANSAS CITY,
MISSOURI INDUSTRIAL HASTEHATER TREATMENT PLANT.
Day
Fathead Minno
24-hour LC50
(% effluent)
/ 1
w - Tr
32
eated
26
2
Sffluei
40
It
32
Daphnia magna - Treated Effluent
24-hour LC50
(t effluent)
lethal units
21
4.8
21
4.8
•
.
3
40
26 .
3.8
32
20
5.0
4
40
;
32
-
5
26
26
3.8
26
22
4-5
6
32
-
26
-
7
26
22
4.5
26
26
3.8
B
21
-
p.ta.
i7
-
9
21
23
4.3
17
21
4.6
10
21
-
23
-
Mean
28
23
4.4
St.
Dev.
7.0
1.0
2.4
0.5
U)
Tests not required at this time period.
-------
TABLE 11. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100%
TEST WATERS DURING TEN CONSECUTIVE DAYS OF ON-SITE
TESTING AT THE MOBAY CHEMICAL, KANSAS CITY, MISSOURI
INDUSTRIAL WASTEWATER TREATMENT PLANT.
Temperature
PH
Dissolved oxygen
(mg/£)
Conductivity
(y mhos/cm)
Hardness (mg/Jl)
Alkalinity (mg/2.)
Chlorine (mg/Jl)
Diluent
'Mean
22
8.4
NA
500
170
120
ND '
water
Range
22-24
8.3-8.5
NA
500-550
160-180
120-130
ND
Treated
.Mecrn
24
7.3
7.6
21,000
NA
NA
ND
ef .fluent
.Range
20-26
7.0-7.4
7.0-8.2
20,000-23,000
NA
NA
ND
Analyses not routinely performed.
ND = non-detectable, e.g. below detectable limits of 0.01 mg/£.
36
-------
TABLE 12. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING ON-SITE
TESTING OF FATHEAD MINNOWS* AND DAPHNIA MAGNAt TO "TREATED
- EFFLUENT" FROM THE MOBAY CHEMICAL, KANSAS CITY, MISSOURI
INDUSTRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT
THE RANGE OF MEASUREMENTS OBSERVED DURING THE TESTING PERIOD.
Dissolved
oxygen
Concen- (mg/i)
tration Hour A. Mo
Effluent/Fathead minnow
-control 0 S..4--9..-4
24 7 . 6-8 . 6
50% 0 7.8-9.0
24 0.4-3.9
21% 0 8.4-9.5
24 2.7-6.5
8.9% 0 8.4-9.6
24 5.0-7.8
Ef f luent/Daphnia magna
control 0 8 . 4-9 . 1
24 7.8-8.6
50% 0 7.7-8.8
24 4.2-6.1
21% 0 8.2-9.1
24 5.9-7.7
8.9% 0 8.2-9.2
24 7.5-8.4
P.M.
«.*-9...S
7.7-8.7
7.4-8.9
0.4-4.5
8.3-9.4
1.3-6.7
8.4-9.5
4.7-8.0
8.4-9.0
7.7-8.7
8.0-8.5
3.7-5.9
8.3-8.9
6.4-7.6
8.4-9.0
7.4-8.5
Conductivity
PH
A.M.
S-.4-3..5
8.2-8.4
7.2-7.5
7.5-7.7
7.4-7.6
7.6-7.8
7.7-7.9
7.8-8.0
8.4-8.5
8.2-8.4
7.2-7.6
8.1-8.2
7.4-7.7
8.2-8.3
7.7-8.0
8.2-8.3
P.M.
^8,3-8.5
8.2-8.4
7.2-7.6
7.6-7.8
7.3-7.7
7.6-7.8
7.6-8.0
7.7-8.0
8 . 3-8 . 4
8.2-8.4
7.2-7.6
8.1
7.4-7.7
8.2
7.8-7.9
8.2
(umhos/cm)
A.M.
SBB-35B
NA11
11000-12000
NA
4500-5400
NA
2200-2600
NA
500-550
NA
11000-12000
NA
5000-5500
NA
2400-2500
NA
P.M.
£TO>-5*0
NA
10000-12000
NA
5000-6000
NA
2100-2700
NA
500-550
NA
10000-12000
NA
5000-5500
NA
2400-2500
NA
*
Fathead minnows tested
for ten consecutive
Daphnia magna tested every other
day, i.e.
days.
five days
Analyses not routinely performed.
37
-------
loo
TABLE 13. LC50 VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS AND DAPHNIA MAGNA EXPOSED TO "UNTREATED INFLUENT" AND "TREATED EFFLUENT" FROM THE
MONSANTO, LULING, LOUISIANA INDUSTRIAL HASTEHATER TREATMENT PLANT.
Day
Fathead Minna
24-hour LCSO
(» influent)
lethal units
/ 1
P
w - Untreate
<1.B
>56
<1.8
>56
2
d Infl
1.0
100
_ _
uent
0.6
170
Fathead Minnow - Treated Effluent
24-hour LCSO
(% effluent)
lethal units
13
7.7
12
8.3
17
5.9
13
7.7
Daphnia roagna - Untreated Influent
24-hour LCSO
(» influent)
lethal units
1.8
56
1.3
77
*
_
Daphnia magna - Treated Effluent
24-hour LCSO
(% effluent)
lethal units
32
3.1
32
3.1
-
-
3
1.5
67
17
5.9
1.0
100
40
2.5
1.2
B3
13
*,7
6.8
120
32
3.1
4
1.2
83
17
5.9
-
-
i.s
67
14
7.1
-
-
5
1.5
67
18
5.6
0.6
170
25
4.0
1.8
56
14
7.1
1.4
71
21
4.8
6
1.0
100
21
4.8
-
-
1.0
100
13
7.7
-
-
7
1.8
56
21
4.8
1.5
67
40
2.5
1.8
56
17
5.9
1.2
83
44
2.3
8
1.8
56
21
4.8
-
-
p. hi.
1.5
67
21
4.8
-
-
9
2.2
45
26
3.8
1.5
67
32
3.1
1.5
67
21
4.8
1.5
67
36
2.8
10
a.ta.
1.8
56
26
3.8
-
-
2.2
45
21
4.8
-
-
Mean
1.5
74
18
6.0
1.3
88
33
3.1
St.
Dev.
0.4
29
4
i-5
0.4
34
7.0
0.7
Tests not required at this time period.
-------
TABLE 14. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURING
TEN CONSECUTIVE DAVS OF ON-SITE TESTING AT THE MON6ANTO, LULtSG, LOUISIANA
INDUSTRIAL WASTEWATER TREATMENT PLANT.
Temperature ( C)
pH
Dissolved oxygen
(mg/£)
Conductivity
(pmhos/cm)
Hardness (mg/fc)
Alkalinity (mg/fc)
Chlorine (mg/fc)
Diluent
Mean
22
8.3
*
NA
500
160
120
NDf
Welter
Range
21-23
8.1-8.5
NA
480-510
160-170
110-120
ND
Treated
Mean
16
8.6 ,
9.7
7900
NA
NA
ND
effluent
Range
12-26
8.4-8.7
8.4-11
6300-9000
NA
NA
ND
Untreated
Mean
30
12
5.9
21000
NA
NA
ND
efflueht
Range
18-44
11-13
0.4-9.4
8300-67000
NA
NA
ND-2.9
Analyses not routinely performed.
[
ND = non-detectable, e.g. below detectable limit of 0.01 mg/fc.
-------
TABLE 15. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOW TO "UNTREATED INFLUENT" AND "TREATED
LUEisT" i'KQH THE MONiiANl'O, LUIJNG, Ii)UlSJ.Artft XNUUSsTKiAi, -HasXE-
WATER TKEA1HEST YmHT.. 'fttSSfi VAirOES HEF8E5KJST T3U5 J1&WGS Of
MEASUREMENTS OBSERVED DURING THE TEN CONSECUTIVE DATS OF ON-SITE
TESTING.
'-V --
Concen-
Dissolved oxygen
(mg/Jl)
tration Hour
Influent
control
10-2. 7%f
4.2-1.2%f
1.8-0.51%f
Effluent
control
32%
14%
5.8%
0
24
0
24
0
24
0
24
D
24
0
24
0
24
0
24
A.M.
8.7-11
7.6-8.6
8.2-11
4.4-9.1
8.8-11
2.7-8.7
8.7-11
2.6-8.1
B-6-13.
7.8-8.8
8.8-11
0.4-7.1
8.8-11
5.4-8.5
8.7-11
6.9-8.7
P.M.
8.5-10
8.0-8.8
7.9-10
6.9-9.1
8.6-10
1.1-9.0
8.6-10
1.6-8.8
8..-7-3.D
8.1-8.7
8.6-11
0.5-7.6
8.8-11
3.9-8.4
8.6-10
6.6-8.8
PH
A.M.
8.1-8.4
8.0-8.4
9.5-11
9.1-11
9.1-10
8.7-10
.8.8-9.7
8.4-9.2
AM-*
8.0-8.5
8.5-8.7
8.4-8.6
8.5-8.7
8.4-8.6
8.4-8.6
8.4-8.5
P.M.
8.0-8.5
8.0-8.2
9.3-12
9.2-11
9.0-11
8.5-11
8.7-10
8.6-10
a^o-s-5
.8-0-8.2
8.5-9.9
8.4-8.6
8.5-9.4
8.4-8.6
8.4-9.0
8.2-8.5
Conductivity
( \i mhos /cm)
A.M.
460-510
*
NA
680-1500
NA .
570-890
NA
520-620
NA
35D-57D
NA
3200-4400
NA
1700-2200
NA
1700-2200
NA
P.M.
460-500
NA
570-2200
NA
550-1100
NA
510-710
NA
460-300
NA
3200-3600
NA
1300-2000
NA
1300-2000
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day. "<»aeu <-•" results or
40
-------
TABLE 16. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE
TESIIHG OF DaPJ
mi& MAGH&
TO "BNTRE1
WATER •EKEATHEST PLANT. THESE "TALOT
MEASUREMENTS OBSERVED DURING THE F3
Concen-
tration
Influent
control
6.5-27%*
2.7-0.75%
1.2-0.32%
Effluent
control
77-50%f
32-21%f
14-8. 9%*
Dissolved oxygen
tmg/fc)
Hour
0
24
0
24
* 0
24
f 0
24
0
24
0
24
0
24
•x.
0
24
A.M.
8.7-9.9
7.7-8.7
8.5-9.6
5.8-8.6
8.7-9.6
4.4-8.4
8.6-9.7
4.6-8.5
B.S-9.S
7.7-8.7
€.4-9.9
4.8-7.6
8.5-9.8
7.0-8.4
8.7-9.8
7.3-8.5
P.M.
8.6-9.5
8.0-8.8
7.9-9.6
7.8-8.9
8.6-9.6
1.8-8.6
8.6-9.7
6.8-8.8
B.7-9.,7
8.1-8.9
8.4-9.7
2.6-7.6
8.6-9.6
6.0-8.6
8.7-9.6
7.5-8.7
5.TED JSFLDEHT* -AHD TB
;, -LODXS1AHA IRDUSTBIA
iS iiKPitESKiW THE RANGE
[VE DAYS OF ON-SITE TE
PH
A.M.
8.0-8.4
8.1-8.4
9.6-11
9.0-9.8
9.2-9.9
8.0-9.0
8.7-9.5
8.1-8.4
B.1-B..4
8.1-8.2
8.5-8.6
8.5-8.6
8.5-8.6
8.5-8.6
8.5-8.6
8.4-8.5
P.M.
8.0-8.4
8.1-8.2
9.8-11
9.2-10
9.2-10
8.5-9.5
8.6-9.4
8.2-8.8
8. 0-1.4
8.1-8.4
8.4-8.6
8.5-8.6
8.5-8.6
8.4-8.6
8.4-8.6
8.4-8.5
EATED
I* WAiTi.-
'OF
STING.
Conductivity
(wmhos/cm)
A.M.
450-550
*
NA
800-1100
NA
600-800
NA
500-600
NA
450^500
NA
4400-7000
NA
2300-3300
NA
1200-1800
NA
P.M.
470-490
NA
700-1400
NA
600-800
NA
500-600
NA
4SO-310
NA
4300-7000
NA
2200-3600
NA
1200-1800
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
-------
TABLE 17. LC50 VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS AND PAfHUIA MAGMA EXPOSED TO "UNTREATED INFLUENT" ADD "TR2ATED EFFLUENT" FROM THE
DIAMOND SHAMHOCX, GREEN BAYOU, TEXAS INDUSTRIAL HASTEHATER TREATMENT PLANT.
Day
/ 1
a.m. I p.m.
1
2
a.m.
p.m.
Fathead Minnow - Untreated Influent
24-hour LC50
<» Influent)
lethal units
>0.25
<400
0.13
770
2.7
37
0.10
1000
Fathead Minnow ~ Treated Effluent
24-hour LC50
It effluent)
lethal units
81
1.2
81
1.2
81
1.2
81
1.2
Daphnia nagna - Untreated Influent
24-hour LC50
(% influent)
• lethal units
0.20
500
<0.057
>1800
t
-
-
Daphnia maqna - Treated effluent
24-hour LC50
(« effluent)
lethal units
32
3.1
40
2.5
~
~
3
a.m.
S.2
19
*
94
1.1
>0.49
<200
40
2.5
p.m.
5.0
20
*
94
1.1
>1.2
<80
40
2.5
4
a.n.
2.1
48
81
1.2
-
-
p.m.
<0.04!
>2200
90°
1.1
-
-
5
a.m.
8.1
12
81.
1.2
S.9
26
40
2.5
p.m.
5.2
19
a
90
1.1
2 4
42
40
2^5
6
a.m.
0.25
<400
a
94
1.1
-
-
p.m.
4.2
24
*
90
1.1
-
-
7
a.m.
3.4
29
81
1.2
1.0
100
40
2.S
p.m.
4.2
24
81
1-2
1.4
71
32
3.1
8
a.m.
0.06
1700
81
1.2
-
-
p.m.
0.06
1700
81
1.2
-
~
.9
a4.2
<24
«0
2.5
p.m.
4.2
24
81
1.2
28
40
2.5
10
o.E.
=0.045
•2200
81
1.2
-
~
p.m.
<0.17
>590
81
1.2
-
-
Mean
3.3
360
84
1.2
130
38
2.6
St.
Dev.
1.4
640
S.O
0.0
i 3
180
3.2
0.2
LCSO valueu calculated by plotting on graph paper.
Tests not required at this time period.
i
-------
TABLE 18. RESULTS OF WA^ER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURING
TEN CONSECUTIVE DAYS OP ON-SlTE TESTING AT THE DIAMOND-SHAMROCK, GREEN
BAYOU, TEXAS INDUSTRIAL WAgfEWATER TREATMENT PLANT.
Temperature (°C)
PH
Dissolved oxygen
(mg/i)
Conductivity
(ymhos/cm)
Hardness (mg/Jl)
Alkalinity (mg/£)
Chlorine (mg/fc)
Diluent
Mean
22
8.2
*
NA
490
160
120
Nbf
water
Range
21-23
81.0-8*5
NA
480-5tJO
160-ieo
110-l2ti
ND
Treated
Mean
9.9
7.8
10
17000
NA
NA
ND
effluent
Range
5.0-16
7.2-8.0
7.8-12
12000-21000
NA
NA
ND
Untreated
Mean
25
7.1
7.3
25000
NA
NA
ND
influent
Range
12-34
2.7-12
3.0-9.4
6000-64000
NA
NA
ND
Analyses not routinely performed.
ND = non-detectable, e.g. below detectable limit of 0.01 mg/£.
-------
TABLE 19. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOWS TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE DIAMOND SHAMROCK, GREEN BAYOU, TEXAS INDUSTRIAL
WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE OF
MEASUREMENTS OBSERVED DURING THE TEN CONSECUTIVE DAYS OF ON-SITE
'. TESTING.
Concen-
tration Hour
Influent
control
10-0. 25%f
4.2-0.10%H
0
24
0
24
h 0
24
Dissolved oxygen
(mg/X.)
A.M.
8.6-11
6.6-8.0
4.1-11
0.5-8.9
7.5-11
3.0-8.5
1, 7-0.043^ 0 ;8-«-Xl
Effluent
control
100%
42%
18%
24
0
24
0
24
0
24
0
24
6.B-8.3
8.0-9.3
7.2-8.3
9 ,..0-12
0.7-5.4
8.5-10
2.2-7.0
8.3-9.6
3.5-7.6
P.M.
8.5-10
7.3-8.7
8.1-10
6.8-8.9
8.7-10
6.7-8.8
8.8-10
7.3-S,*
8.5-9.6
7.4-8.5
3.0-11
0.6-5.9
8.7-10
1.6-7.4
8.7-10
3.2-8.3
PH
A,M.
7.9-8.5
7.9-8.3
7.0-11
7.1-9.9
7.5-9.5
7.5-8.6
7,5-9,2
7,3-8,4
7.8-8.4
7.8-8.4
7,4-7,9
7 . 2-7 .B
7.5-8.1
7.3-7.9
7.6-8.2
7.4-8.0
*,M,
8.0-8.4
7.2-8.4
6.5-12
6.9-11
7.0-11
7.3-10
7.5^-B
1,5-9,, 4
8.1-8.4
7.9-8.4
7,4-8,3
7.3-7.3
7.6-8.3
7.5-7.9
7.7-8.3
7.5-7.9
Conductivity
(jnnhos/cin)
A.M.
450-550
NA*
470-2900
NA
460-1300
NA
4*0-800
SA
470-550
NA
19000-22000
NA
9000-10000
NA
4500-7000
NA
P.M.
460-500
NA
500-6500
NA
480-3200
NA
460-1400
IJA
450-500
NA
20000-22000
NA
lOOOOr-11000
NA
4500^-7000
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
44
-------
TABLE 20. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF DAPHNIA MAGNA TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE DIAMOND SHAMROCK, GREENS BAYOU, TEXAS INDUS-
TRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE
RANGE OF MEASUREMENTS OBSERVED DURING THE FIVE DAYS OF ON-SITE
TESTING .
Concen-
tration
Influent
control
10-0.32%f
1.8-0.13%
0,35-0^9
Effluent
control
77-50%
32-21%
14-8.9%
Dissolved oxygen
(mq/1)
Hour
0
24
0
24
* 0
24
»f 0
.24
0
24
0
24
0
24
0
24
A.H.
8.6-9.1
6.5-8.6
8.7-9.2
6.7-8.7
8.7-9.0
6.6-8.6
8.5-9,1
fi,€-8-7
8.. 5-8. 9
6.6-8.8
9.0-10
4.5-3,1
8.8-9.4
6.3-8.5
8.6-9.0
6.6-8.8
P.M.
8.3-8.9
7.9-8.9
7.5-9.0
6.9-8.8
8.3-8.9
7.9-8.8
8-3-8.9
7-9-3.8
8.3-8.8
7.9-8.7
8.9-10
3-9-7.6
8.7-9.6
6.6-8.5
8.4-9.0
7.4-8.7
PH
A.M.
7.8-8.4
8.3-8.4
7.4-8.2
7.9-8.3
7.8-8.2
8.2-8.4
7.8-8.2
8. .2-3.. 4
7.7-8.3
8.1-8.5
7.3-8.0
7.6-8-0
7.3-8.1
7.8-8.1
7.4-8.2
7.9-8.2
Conductivity
(pmhos/cm)
P.M.
8.1-8.5
8.2-8.4
6.9-12
7.5-10
7.4-10
7.8-9.5
7.8-9.5
8.0-8.8
8.0-8.5
8.1-8.4
7.5-8.0
7.6-8-0
7.6-8.1
7.8-8.0
7.7-8.2
7.9-8.1
A.M.
470-550
NA*
500-1700
NA
500-850
NA
500-600
m
470-500
NA
11000-17000
NA
5500-8000
NA
2500-3800
NA
P.M.
480-500
NA
600-4500
NA
490-1800
NA
470-750
NA
460-600
NA
11000-1700C
NA
5000-8000
NA
2500-3700
NA
Analyses not routinely performed.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
45
-------
TABLE 21. LCSO VALUE AND LETHAL UNITS FOR FATHEAD MINNOWS EXPOSED TO TWO "TREATED INFLUENTS* AND A "TREATED EFFLUENT" FROM THE DUPONT LAPORTE
TEXAS INDUSTRIAL WASTEWATER TREATMENT PLANT. '
Day
Fathead Minno
24-hour LC50
(» influent)
lethal units
/ J
A ra
1
v - Treated
26
3.8
26
3.8
2
:nflue
32
3.1
*
it
40
2.5
Fathead Minnow - Treated Influent***
24-hour LCSO
(a influent)
lethal units
0.54
180
0.84
120
0.65
150
0.80
120
Fathead Minnow - Treated Effluent
24-hour LCSO
(« affluent)
42
42
52
92
3
32
3.1
0.84
120
42
<8.9
>11
0.84
120
52
4
14
7.1
n
42
u
42
5
32
J.I
32
>50
<2.0
42
6
<8.9
>11
34
8.9
11
42
7
32
3.1
42
11
9-1
34
8
40
2.5
42
32
3.1
43
9
<8.9
>11
42
•
<8.9
>11
42
10
26
3.8
42
<8.9
>11
34
Mean
27
4.5
0.75
140
42
St.
Dev.
10
2.8
0.13
27
5.6
0.3
Process wastewater Stream til - posticide/non-pesticide main process water.
Process wastewater Stream «2 - Maneb/He thorny l production prior to chemical oxidation treatment.
Testing terminated due to process shutdown.
-------
TABLE 22. LCSO VALUES AND LETHAL. UNITS FOR DAPHNIA IJACNA EXPOSED TO TWO "TREATED INFLUENTS" AND A "TREATED EFFLUENT" FROM THE DUPONT, LAPORTE, TEXAS
INDUSTRIAL HASTEWATER TREATMENT PLANT.
Day
Daphnia magna
24-hour LCSO
(% influent)
letnal units
/ 1
r=nP...
1
- Treated I
>21
<4.8
>21
<4.8
2
nf luen
_t
*
t
_
Daphnia magna - Treated Influent^
24-hour LCSO
(t influent)
lethal units
0.16
620
0.17
S90
_
^_
Daphnia magna - Treated Effluent
34-hour LCSO
(» effluent)
lethal units
2.4
42
<4.2
>24
_
_
3
16
6.2
0.1S
670
3.3
30
<9.0
>11
O.OB
1200
2.8
36
4
-
-
-
-
/
-
S
10
10
I
o.es
120
>so
<2.0
4.2
24
6
-
-
-
-
7
40
2.S
4.B
21
36
2.8
S.2
19
8
-
-
-
-
9
<40
>2.5
6.3
16
40
2.S
9.3
11
10
-
-
-
-
-
-
Mean
28
4.8
O.K
770
4.4
35
St.
Dev.
14
3.3
0.04
310
2.S
31
Process wastevater Stream »1 - pesticide/non-pesticide Bain process water.
Tests not required at this time period.
n
Process wastewater Stream «2 - Maneb/methomyl production prior to chemical oxidation treatment.
Testing terminated due to process shutdown.
-------
TABLE 23. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURING
TEN CONSECUTIVE DAYS OF ON-SITE TESTING AT THE DUPONT, LAPORTE, TEXAS IN-
DUSTRIAL WASTEWATER TREATMENT PLANT.
Temperature (°C)
PH
Dissolved oxygen
Conductivity
(ymhos/cm)
Hardness (mg/£)
Alkalinity (mg/£)
Chlorine (mg/£)
Diluent
Mean
23
8.2
NA
500
170
120
ND11
water
Range
22-24
8.1-8.5
NA
490-510
160-180
120-140
ND
Treated
Mean
16
8.0
8.8
9800
NA
NA
ND
effluent
Range
11-21
7.6-8.8
7.3-11
6000-11000
NA
NA
ND
Treated
Mean
34
8.1
6.4
8700
NA
NA
20
*
influent
Range
30-39
2.7-12
2.7-10
3900-21000
NA
NA
0-44
t
Process wastewater stream #1 - pesfcicide/non-pesticide main process water.
h
Analyses not routinely performed.
1
ND = non-detectable, e.g. below detectable limit of 0.01 mg/JZ..
-------
TABLE 24. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOWS TO TWO "TREATED INFLUENTS'" FROM THE
DUPONT, LAPORTE, TEXAS INDUSTRIAL WASTEWATER TREATMENT PLANT.
THESE VALUES REPRESENT THE RANGE OF MEASUREMENTS OBSERVED DURING
THE TEN CONSECUTIVE DAYS OF ON-SITE TESTING.
Concen-
tration
*
Influent
control
50$
21%
8.9%
Influent11
control
2.0%
0.84%
0.36%
Dissolved oxygen
(mg/i)
Hour A.M.
0
24
0
24
0
24
0
24
,8
0
24
0
24
0
24
0
24
7
6
7
0
8
0
7
2
8
6
8
0
.9-9.7
.6-8.6
.5-8.9
.2-8.3
.0-9.5
.4-8.6
.4-9.7
.8-8.6
.5-9.3
.6-8.6
.5-9.5
.5-8.0
8.7-9,6
1
8
6
.5-8.3
.6-9.5
.1-8.6
8
7
6
0
8
0
8
0
9
6
9
1
9
3
9
5
P.M.
.0-9.5
.1-8.4
.7-9.5
.6-8.2
.1-9.5
.6-8.4
.1-9.4
.9-8.0
.10-9.1-
.6-7.6
.0-9.3
.1-1.2
.J.-9.2
.7-3.9
.0-9.2
.9-6.3
8.
8.
7.
7.
7.
7.
8.
7.
8.
8.
8.
8.
7.
8.
8.
A.M.
2-8.4
1-8.4
7-12
6-12
9-12
7-11
1-11
8-11
3-8.4
1-8.4
4-9.0
8.3
4-8.8
9-8.3
3-8.6
2-8.4
PH
P.M.
8.2-8.
8 . 1-8 .
3.7-10
3.5-10
5.5-9.
5.7-9.
7.0-9.
7.3-9.
8.3^-8.
8.1-8.
8.9-9.
8.2-8.
8.6-8.
8.0-8.
8.4-8.
5.9-6.
Conductivity
(vmhos/cm)
5
3
7
6
2
1
5
2
1
4
9
2
6
3
A.M.
500
NA*
2100-9000
NA
1200-4000
NA
800-1900
NA
500
NA
500-700
NA
500-600
NA
500
NA
P.M.
500
. NA
2000-5000
NA
1200-2700'
NA
700-1500
NA
500
NA
650-700
NA
550-600
NA
500-600
NA
Treated influent Stream #1 - pesticide/non-pesticide main process water.
Analyses not routinely performed.
Treated influent Stream §2 - Maneb/Methomyl production prior to chemical
oxidation treatment.
Testing terminated after day 3 due to process shutdown.
49
-------
TABLE 25. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE TESTING OF
FATHEAD MINNOWS TO "TREATED EFFLUENT" FROM THE DUPONT, LAPORTE, TEX&3 INDUS-
TRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE RANGE OF
MEASUREMENTS OBSERVED DURING THE TEN CONSECUTIVE DAYS OF ON-SITE TESTING.
on
o
Concen-
tration
Effluent
control
100%
42%
18%
Dissolved oxygen
(mg/Jl)
Hour
0
24
0
24
0
24
0
24
8
6
7
0
8
0
8
3
A.M.
.4-9.6
.7-8.3
.8-11
.4-r4.0
.3-9.7
.8-6.3
.4-9.7
.0-7.1
8
6
7
0
8
1
8
3
P.M.
.4-9.4
.7-8.0
.9-10
.9-4.9
.5-10
.8-6.8
.5-9.6
.3-7.5
A.M.
8.2-8
8.2-8
7.8-8
7.6-7
7.9-8
7.6-8
8.0-8
7.6-8
PH
.5
.4
.1
.9
.2
.1
.3
.1
P.M.
8.2-8
8.1-8
7.8-8
7.6-7
7.9-8
5.7-8
8.0-8
7.8-8
.5
.4
.2
.8
.2
.1
.3
.2
Conductivity
(pmhos/cm)
A.M.
500
NA
10000-120000
NA
4700-6000
NA
2400-3000
NA
P.M.
500
NA
10000-12000
NA
5000-6000
NA
2500-2900
NA
Analyses not routinely performed.
-------
TABLE 26. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF DAPHNIA MAGNA TO TWO "TREATED INFLUENTS" FROM THE
DUPONT, LAPORTE, TEXAS INDUSTRIAL WASTEWATER TREATMENT PLANT.
THESE VALUES REPRESENT THE RANGE OF MEASUREMENTS OBSERVED DURING
THE FIVE DAYS OF ON-SITE TESTING.
Concen-
. tration
*
Influent
control
50-21%n
21-8. 9%n
8.9-4.2%n
Influent8 '
control
0.32-0.21%
0.13-0.09%
0.06-0.04%
Dissolved oxygen
(mq/fc)
Hour
0
24
0
24
0
24
0
24
§
0
24
n o
24
n o
24
n o
24
A.M.
8.5-9.3
7.8-8.7
7.7-8.9
1.8-8.3
8.3-9.3
2.9-8.4
8 .5-9.3
4.8-8.3
8.8-8.9
7.1-7.8
8.8-9.0
7.6-7.8
8 . 7-8 . 8
6.9-8.0
8.7-8.8
7.8-8.0
P.M.
8.7-9.1
7.6-9.4
7.7-9.4
1.3-9.2
8.3-9.7
2.9-9.0
8.6-9.3
2.6-8.1
8.4-8.5
7.8-8.0
8.5-8.7
7.2-7.9
8.6
7.9-8.1
8.5-8.6
7.8-7.9
PH
A.M.
8.2-8.5
8.2-8.4
7.9-12
7.7-12
8.0-12
7.7-11
8.1-11
7.8-10
8.3-8.5
8.1-8.4
8.5-8.6
8.2-8.3
8.4-8.5
8.2
8.3-8.5
8.2-8.4
P.M.
8.2-8.5
8.2-8.4
3.7-9.2
3.5-8.0
5.6-8.9
5.7-8.1
7.3-8.7
7.8-8.1
8.3-8.5
8.1
8.4-8.6
8.0-8.2
8.4-8.6
8.1-8.2
8.3-8.5
8.1-8.2
Conductivity
(pmhos/cm)
A.M.
500
NAf
1100-9500
NA
950-3900
NA
600-1700
NA
410-500
NA
430-550
NA
490-500
NA
500
NA
P.M.
500
NA
1400-5500
NA
750-2700
NA.
650-1500
NA
480-500
NA
490-500
NA
480-500
NA
450-500
NA
n
Treated influent Stream II - pesticide/non-pesticide main process water.
Analyses not routinely performed.
Treated influent Stream f2 - Maneb/ Methomyl production prior to chemical
oxidation treatment.
I
Testing terminated after day 3 due to process shutdown.
Test concentrations varied during testing and were based on results of
toxicity data from the previous day.
51
-------
TABLE 27. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE TESTING OF
DAPHNIA MAGNA TO "TREATED EFFLUENT" FROM THE DUPONT, LAPORTE, TEXAS INDUS-
TRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT THE:RANGE OF
MEASUREMENTS OBSERVED DURING THE FIVE DAYS OF ON-SITE TESTING.
ui
to
Concen-
tration
Effluent
control
21%
8.9%
4.2%
Dissolved oxygen
(rag/ A)
Hour
0
24
0
24
0
24
0
24
8
7
8
5
8
6
8
6
A.M
.4-8
.1-8
.4-9
.9-6
.4-9
.2-7
.3-9
.6-7
•
.9
.3
.4
.8
.3
.1
.3
.6
P.M.
8.0-9.1
6.5-7.9
8 . 4-9 . 3
5.2-7.3
8.4-9.3
5.8-7.7
8.2-9.0
5.9-7.4
A.M
8.3-8
8.1-8
8.0-8
7.8-8
8.1-8
7.8-8
8.2-8
7.9-8
PH
P.M.
.5
o4
.2
.0
.4
.1
.4
.2
8.2-8
8.0-8
7.9-8
7.8-8
8.0-8
7.9-8
8.1-8
7.9-8
.5
.3
.2
.0
.3
.1
.4
.2
Conductivity
(ymhos/cm)
A.M.
460-500
*
NA
2000-3500
NA
1200-1700
NA
800-1000
NA
P.M.
490-500
NA
2200-3000
NA
1400-1600
NA
950-1000
NA
Analyses not routinely performed.
-------
TABLE 28. LC50 VALUES AND LETHAL UNITS FOR FATHEAD MINNOWS AND DAPHNIA MAGMA EXPOSED TO "UNTREATED INFLUENT" AMD "TREATED EFFLUENT" FROM THE
INTERNATIONAL PAPER, GEORGETOWN, SOUTH CAROLINA INDUSTRIAL HASTEHATER TREATMENT PLANT.
Day
Fathead Minno
24-hour LC50
(» influent)
lethal units
/ 1
w - Untreate
52
1.9
34
2.9
2
d lufl
42
uent
34
Fathead Minnow - Treated Effluent
24-hour LC50
(» effluent)
lethal units
>100
0
•100
0
>100
0
>100
0
Daphnia magna - Untreated Influent
24-hour LCSO
(t influent)
lethal units
>65
<1.5
>«<65
>1.5
<4.2
*
-
Daphnia tnagna - Treated Ef fluent T
24-hour LCSO
(t effluent)
lethal units
>100
0
>100
0
-
-
3
34
>100
0
55
1.8
>100
0
42
>100
0
61
1.2
>100
0
4
42
>100
0
-
-
52
>100
0
-
-
5
52
>loo
0
.77
1. 3
>100
0
42
>100
0
73
1 4
>100
0
6
42
>100
0
-
-
23
>100
0
-
-
7
42
>100
0
77
1 3
>100
0
42
2.4
>100
0
>65
<1 5
>100
0
8
23
>10O
0
-
-
21
>100
0
-
-
9
42
2.4
>100
0
41
>100
0
34
2.9
>100
0
42
>100
0
10
27
3.7
>100
0
-
-
<12
>8.3
>100
0
-
-
Hean
3B
2.8
•100
0
£4
•10O
0
•St.
Dev.
9.5
0.8
17
U)
Tests not required at this time period.
No mortalities observed in any test concentration
-------
TABLE 29. RESULTS OF WATER CHEMISTRY ANALYSES CONDUCTED ON 100% TEST WATERS DURING
TEN CONSECUTIVE DAYS OF ON-SITE TESTING AT THE INTERNATIONAL PAPER, GEORGE-
TOWN, SOUTH CAROLINA INDUSTRIAL WASTEWATER TREATMENT PLANT.
-Diluent water
Temperature (°C)
PH
Dissolved oxygen
(mg/£)
Conductivity
(ymhos/cm)
Hardness (mg/£)
Alkalinity (mg/SL)
Chlorine (mg/£)
Mean
22
8.3
NA
500
170
120
NDf
Range
21-23
8.2-8.4
N*
500
170-180
110-130
ND
Treated
Mean
21
7.5
4.3
2000
NA
NA
ND
effluent
Range
17-23
7.1-7.6
2.5-5.2
1800-2100
NA
NA
ND
Untreated influent
Mean
37
7.1
3.0
2200
NA
NA
ND
Range
35-41
6.1-8.6
0.4-4.9
1300-2600
NA
NA
ND
Analyses not routinely performed.
I-
Non-detectable, e.g. below detectable limit of 0.01 mg/Jl.
-------
TABLE 30. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF FATHEAD MINNOWS TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE INTERNATIONAL PAPER, GEORGETOWN, SOUTH CAROLINA
INDUSTRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT
THE RANGE OF MEASUREMENTS OBSERVED DURING THE TEN CONSECUTIVE DAYS
OF ON-SITE TESTING.
Concen-
tration
Influent
control
65%
27%
12%
Effluent
control
100%
42%
18%
Dissolved oxygen
(mq/i)
Hour
0
24
0
24
0
O A.
0
24
0
24
0
24
0
24
0
24
A.M.
8.7-9.3
7.8-8.6
4.7-7.5
0.3-0.6
7.3-8*5
1ft-»A A
8.3-9.2
4.1-6,2
8 .-6-9 . 1
8.1-8,6
1.5-5.4
0.3-0.8
6.3-7.7
2.5-4.0
7.9-8.4
5.8-6.7
P.M.
8.8-9.2
8.1-8.5
3.9-7.9
0.2-1.3
7.4-8.5
Oe_c c
* 3"" j • 3
812-9.0
4,5-7.0
•B.-8-9.-2
8.0-8.6
2.5-4.0
0.3-0.8
6.8-7.5
2.6-4.2
8.0-8.8
6.3-7.3
PH
A.M.
?
8.2-8.3
8.2-8.3
6.7-8.1
6.8-7.7
7.1-8.1
7 . 4-8 . 2
7.4-7.9
6.2-8.3
8.2-8.3
7.2-7.6
7.3-7.6
7.4-7.8
7.4-7.6
7.8-8.0
7.6-7.8
Conductivity
(umhos/cm)
P.M.
7.4-8.3
7.7-8.2
6.7-8.4
6.9-7.8
7.2-8.1
7.6-8ll
7.6-7.8
8.0-8.3
8.1-8.2
7.0-7.6
7.2-7.5
7.2-7.7
7.3-7.6
7.2-7.9
7.4-7.8
A.M.
500
*
NA
1200-1600
NA
750-980
Mi.
NA
600-700
NA
500
NA
1900-2000
NA
1100-1200
NA
800-850
NA
P.M.
500
NA
1100-1500.
NA. .
750-900
MA
NA
600-700
NA
500
NA
1800-2100
NA
1100-1200
NA
700-800
NA
Analyses not routinely performed.
.5.5.1
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TABLE 31. RESULTS OF WATER CHEMISTRY ANALYSES PERFORMED DURING THE ON-SITE
TESTING OF DAPHNIA MAGNA TO "UNTREATED INFLUENT" AND "TREATED
EFFLUENT" FROM THE INTERNATIONAL PAPER, GEORGETOWN, SOUTH CAROLINA
INDUSTRIAL WASTEWATER TREATMENT PLANT. THESE VALUES REPRESENT
THE RANGE OF MEASUREMENTS OBSERVED DURING THE FIVE DAYS OF ON-SITE
TESTING.
Dissolved oxygen
Concen-
tration
Influent
control
100-65%t
42-27%t
;18-12%t
Effluent
control
100%
42% .
18%
(mg/i)
Hour
0
24
0
24
0
24
0
24
0
24
0
24
0
24
0
24
A.M.
8.6-9.2
8.0-8.5
4.6-7.1
0.8-1.8
7.5-8.4
1.9-5.9
8.2-9.2
5.0-7.6
8.. 6-9.0
8.0-8.4
2.6-3.9
2.3-3.5
6.9-7.4
5.3-6.9
7.9-8.5
7.4-7.9
P.M.
8.5-9.0
8.3-8.7
2.6-7.0
0.7-2.4
6.7-8.1
3.6-5.5
7.8-8.4
6.1-7.5
8.5-9.0
8.3-8.6
2.6-3.9
2.6-3.7
6.7-7.2
5.6-6.7
8.0-8.4
7.4-7.9
PH
A.M.
8.2-8.3
8.2
6.8-7.4
7.0-7.8
7.2-7.7
7.7-7.9
7.6-8.0
7.8-8.0
8.2-8.3
8.2
7.4-7.6
7.8
7.6-7.8
7.9-8.0
7.8-7.9
8.0-8.1
P.M.
8.2-8.3
8.2
6.5-7.7
7.2-8.0
7.1-7.9
7.5-7.9
7.5-8.0
7.8-8.0
8.2-8.3
8.2
7.4-7.6
7.7-7.8
7.6-7.7
7.8-7.9
6.7-7.9
7.5-8.1
Conductivity
(p mhos/cm)
A.M.
500
NA*
1500-1900
NA
950-1100
NA
700-800
NA
500
NA
1800-2100
NA
1100
NA
750-800
NA
P.M.
500
NA
1100-1800
NA
800-1000
NA
600-750
NA
500
NA
1700-2000
NA
IO'00-IIOO
NA
700-800
NA
Analyses not routinely performed.
Test concentrations varied during testing .and were based on results of
toxicity data from the previous day.
56-
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APPENDIX
EG&G MASON RESEARCH INSTITUTE
Modified Protocol for Salmonella/Mammalian Microsome
Plate Incorporation Mutagenesis Assay
Media
Top agar is initially prepared with 13.4 g/liter Difco Bacto
Agar and 8.4 g/liter NaCl. After autoclaving, the molten agar
is distributed in 100 mi aliquots into sterile bottles where it
is stored at room temperature. Immediately before its use in
the mutagenesis assay, the top agar is melted and supplemented
with 10 mJl/100 mi agar of a sterile solution containing 0.8 mM .
L-histidine and 0.8 mM biotin (1XSA). Twenty-five ml of sterile
deionized water is added per 100 mi top agar when it is used in
assays without metabolic activation. This insures that final
top agar and amino acid supplement concentrations are the same on
plates with or without metabolic activation.
Top agar for cell titers is initially prepared with 8 g/
liter Difco Bacto Agar and 5 g/liter NaCl. After autoclaving,
the molten agar is distributed in 100 mi aliquots. Immediately
before its use, the top agar is melted and supplemented with 10
mi/IQO mi agar of a sterile solution containing 5.0 mM L-histi-
dine and 0.5 mM biotin (10XSA).
Bottom agar is the Vogel-Bonner minimal medium E described
by Ames.
Nutrient broth used for growing overnight cultures of the
tester strains contains 25 g per liter of Nutrient Broth No. 2
(Oxoid).
Storage and Preparation of Tester Strains
All tester strains are stored in liquid nitrogen, and fresh
cultures are inoculated directly from these frozen stocks. Broth
cultures are grown overnight at 37°C with shaking. If necessary,
the cultures are then centrifuged and resuspended in their cul-
ture medium to give an appropriate final cell concentration.
Each culture is routinely checked for crystal violet sensitivity
and ampicillin resistance before use in the mutagenesis assay.
57
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Tox-icity Determination of Test Compounds
Each test compound is checked for toxicity to the tester
strains up to a concentration of 1.0 m£/plate. Eight serial
ha-tff-log dilutions of the test compound are plated with TA100 on
minimal agar plus 10XSA and on minimal agar plus 1XSA. Equal
numbers of cells are seeded on each plate in the presence of test
compound. The percent survival of an appropriately diluted TA100
culture on the 10XSA supplemented plates is determined by com-
paring numbers of colonies on the solvent control with those on
the plates containing test compound. Toxicity of the 1XSA sup-
plemented plates is detectable by a decrease, in the number of
revertant colonies occurring per bacterial lawn. The highest
concentration of test compound used in the broad range mutagenesis
assay is that which gives approximately 35% survival on the 10XSA
supplemented plates and a detectable reduction of spontaneous
revertants on the 1XSA supplemented plates. If no toxicity is
apparent, then the highest concentration used in the broad range
mutagenesis assay is 1.0 mJl/plate.
Plating Procedures for Mutagenesis Plate Incorporation Assay
Routinely, the test compound is thawed and filtered imme-
diately before its use in the mutagenesis assay. Five doses of
the compound are first plated with all five tester strains (TA98,
TA100, TA1535, TA1537, TA1538) with metabolic activation, after
which they are immediately plated on all tester strains without
metabolic activation. All positive controls, solvent controls,
and test compound dilutions are plated in triplicate. Without
metabolic activation, 50 u£ of tester strain and 1.0 mZ of sol-
vent or test compound are added to 1.5 m£ of molten top agar at
45°C. With metabolic activation, 50 uH of tester strain, 1.0
m£ of solvent or test compound, and 0.5 mfc of S-9 mix are added
to 1.0 of molten top agar at 45°C. After vortexing, the mixture
is poured onto the surface of 25 m& of bottom agar contained in
a 15 x 100 mm plastic per.tri dish.
Positive Controls
Positive controls are run with each day's assays. 2-Amin-
oanthracene (2AA) is plated at 0.4 ug/plate with metabolic acti-
vation on strains TA98, TAlOO, and TA1538. Propane sultone (PS)
is plated at 0.02 uA/plate without activation on TA1535, and 9-
aminoacridine (9AAD) is plated at 75 ug/plate on TA1537 without
activation.
Tester Strain Titers
Tester strain titers are determined by viable count assays
on 10XSA supplemented minimal agar plates. The number of cells
plated per plate is reported on form WL-59 in the final report.
58
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Preparation and Storage of Liver Microsomal Enzymes
Liver microsomal enzymes are routinely prepared from male
Sprague-Dawley rats that have been injected with Arochlor 1254 at
mg/kg. The Arochlor is diluted in corn oil to a concentration
of 200 mg/ml. Five days after their i.p. injection with the
Arochlor, the rats are sacrificed by decapitation, and their
livers are excised. The rats are denied access to food for 12
hour immediately preceding sacrifice.
The preparation of the microsomal enzyme fraction is carried
out with sterile glassware and solutions at 0-4°C. The liver
from each rat is excised and placed in 20 m of 0.15 M KC1 con-
tained in a pre-weighed beaker. After weighing the liver, it is
transferred to another beaker containing 3 volumes of 0.15 M KC1
(3 ml/g of wet liver) where it is minced with sterile scissors.
The minced liver is homogenized in a Potter-Elvehjen apparatus
with a teflon pestle. The homogenate is centrifuged at 9000 x g
for 10 minutes in the SS-34 rotor of a Sorvall SS-3 centrifuge.
The supernatant (referred to by Ames at the S-9 fraction) is de-
canted, and small aliquots are distributed into freezing ampules
which are stored in liquid nitrogen.
One ml of the microsomal enzyme reaction mixture (S-9 mix)
which is added to the soft agar overlay contains the following
components:
S-9 0-.05 ml
0.4 M MgCl2 0.02 ml
1.65;M KC1 0.02 mi
0.04 M NADP • 0.10 ml
0.05 M Glucose-6-phosphate 0.10 ml
1.00 M NaH2PO.,, pH 7.4 0.10 ml
H20 0.61 ml
1.00 ml
Evaluation of Mutagenesis Assay Data
For a compound to be considered positive, it must cause a
doubling in the observed reversion index of at least one tester
strain. The increase in reversion index must be accompanied by
a dose response to increasing concentrations of the test compound.
59
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