ML17223B311

Final Rept Chronic Toxicity of Betz Clam-Trol CT-1 in Mysidopsis Bahia, Including Betz Lab,Inc Environ Info on Clam-Trol CT-1 for Macrofouling Control

ML17223B311
Person / Time
Site:	Saint Lucie
Issue date:	07/31/1991
From:	AQUA SURVEY, INC.
To:
Shared Package
ML17223B307	List: ML17223B306 ML17223B308 ML17223B309 ML17223B310 ML17223B311 ML17309A663
References
BR91-1513A, NUDOCS 9111140237
Download: ML17223B311 (56)

Text

ATTACHMENT7 RECUR SURVEY, INC.

FINAL REPORT THE CHRONIC TOXICITY OF BETZ CLAM-TROL CT-1 IN QYSIDOPSIS BAHIA JULY 31, 1991 BR91-1513A JOB 091- 089A 499 Point Breeze Road ~ Flemington, New Jersey 08822 ~ Telephone (908) 788-8700 FAX (908) 788-9165 9111140237 911101 P0R A000K 05000335 P PDR

RQUFI SURVEY, INC.

STUDY TITLE Clam-Trol~ CT-1 Chronic Bioassay with Data Re uirement Author York Terrell Stud Com leted o July 25, 1991 Performin Laborator Aqua Survey, Inc 499 Point Breeze Road Flemington, New Jersey 08822 Labo ator Pro'ect I.D 91-089A 1 of 12 0

alf gfc~ e Qocc + "-fe~;fl("tI n pew .e'~ey

~

~ Ri,

<<r.Y p g ~ w + Pc r n PpUr e '

!gal >np, Qp ~

~ ~

RECUR SURVCY, IN(.

STATEMENT OF NO CONFIDENTIALITY CLAIMS No claim of confidentiality is made for any information contained in this study on the basis of its falling within the scope of FIFRA Section 10 (D) (1) (A), (B) or (C) .

BETZ LABORATORIES, INC.

4636 Somerton Road Trevose, PA 19047

r. Larr L o s Company Agent Date A atic Toxicolo ist Laborator Mana er Title Signature ASI Project Number: 91-089A 2 of 12

RC)UR SURVEY, INC.

THE ACUTE TOXICITY BIOASSAY OF CLAM-TROL, CT-1

" 5" This study does not meet the requirements for 40 CFR Part 160.

Submitter: Betz laboratories, Inc.

4636 Somerton Road Trevose, PA 19047 Signature:

Date Sponsor: Betz Laboratories, Inc.

4636 Somerton Road Trevose, PA 19047 Signature:

Date Study Director:

ork Terrell D e 3 of 12 d~9.-" Ir.~ Gree..o 9"cc ~ =,ter In@'"o i',-v

~O Gcx 8'C.=i ~ Bc cocci.ge!" ':o o P< ~

~ ->egg t lp

HDUR SURVCY, INC.

Summary A sample of Betz Clam-Trol, CT-1, received from Betz Laboratories, Inc., Somerton Road, Trevose, PA 19047 on March 21, 1991, was tested for potential chronic toxicity to the saltwater static conditions with daily renewal of the test solutions.

Reported final test endpoints are a Nominal No Observed Effects Concentration (NOEC) of 1.25 ppm, a Nominal Lowest Observed Effects Concentration (LOEC) of 2.5 ppm and a 168 hour0.00194 days <br />0.0467 hours <br />2.777778e-4 weeks <br />6.3924e-5 months <br /> Lethal Concentration (LC50) of 1.37 ppm.

4 of 12

<<cQ pq<<~<< -'!:----e Roc@ ~ F'~p <<r;<<-'.<<=A Nev"

RECUR SURVEY, lNC.

I. Objective The objective of this study was to determine the chronic toxic effects of Betz CT-1 in a marine/estuarine aquatic environment with regard to invertebrates. The measure of potency is that concentration, in water, which produces a significant toxic response in the exposed animals after 168 hours0.00194 days <br />0.0467 hours <br />2.777778e-4 weeks <br />6.3924e-5 months <br /> (7 days).

II. Test material Source: Betz Laboratories, Inc.

Somerton Road Trevose, PA 19047 Name: Clam-Trol~, (CT-1) 4 Solubility in Water: 100 Date Received: March 21, 1991 Amount Received: 1 pint III. Materials and Methods A. Method The method employed was a modification of the method outlined by The United States Environmental Protection Agency, Environmental Monitoring Systems Laboratory, EPA-600/4-87/027 May 1988:

of the Appendix.)

B. Test Organisms

~Secies The test species, representing a typical saltwater 5 of 12 "v~ >3~('. "fe8ze igocc ~ 'Fiewif;gic'A .v'e+'6'cey '&~822 ~ . '~ ".<~,~ 8j r" ~

PQ S",:x 8.C~9 ~ Bo;-A P~oi>i-.e ).cps:<<:~,c 766.-' ='e" < e

FlgUR SURVEY, INC.

Size A e Ph sical Condition Animals used for this test were 7 day old mysid shrimp.

Animals appeared to be healthy and in good physical condition.

Source Acclimation All'animals utilized in this test were from a stock culture maintained at this laboratory. The animals were acclimated to test diluent for 48.0 hours0 days <br />0 hours <br />0 weeks <br />0 months <br /> at a moderate changeover rate, prior to testing.

C. Test System Source of Di ution Water Water utilized for this test was filtered (0.45 microns) natural seawater obtained from Manasquan River Inlet, Manasquan, New Jersey.

The test temperature was 26 +/- 0.54 C Test vessels The test vessels were 250 ml, glass beakers containing 200 ml of test solution at a depth of 6 cm.

Photo eriod A 16-hour light/8-hour dark photoperiod with a 30 minute transitional period was utilized.

Loai~g Each test chamber contained 5 organism in 200 ml of test solution.

6 of 12 7 pgj~f ~[ep=. QocQ ~ Pie~lq< <oq f'<ew .ei y

~ ' c.'- c:-o ~ < 8) ".E

~O Bcx c.C5v ~ Ba cn Rouge ~.cu~sicna C68=' ':.,-.-.o-" -. "-."".'.:,~-".-.a:"-'

HOUR SURVEY, lNC.

D. Test Design Test Levels A range finding test was conducted separately. Four concentrations of the test material were evaluated using 10 mysids per treatment. An LC50 determination with 5 concentrations spanning the appropriate range was evaluated using 1 replicate of 10 organisms each per treatment. The definitive test was conducted using 5 organisms in each of 8 replicates at each of 5 treatment levels.

Control A control treatment was run concurrently with the test substance treatments using the same dilution water and the same number of organisms as per the test concentrations.

Test Initiatio Testing was conducted under static conditions with daily renewal of the test solutions. An initial test material stock solution of 1,000 ppm was prepared with deionized water. Test solutions were prepared from the stock solution and then dispensed into the test chambers. The test was initiated when, within 30 minutes of test solution preparation, the test organisms were transferred to the test chambers.

The range-finding test spanned 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> and was conducted using 10 organisms at each of four treatment levels: O.l, 1.0, 10 and 100 ppm.

Test results indicated a toxic response between 1.0 and 10.0 ppm. Exposure at the lower concentration level resulted in no mortality after 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br />, while exposure at the higher level resulted in 1004 mortality after 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br />.

The 96-hour LC50 determination was conducted using 10 organisms in one replicate at each of 5 treatment levels: 0.625, 1.25, 2.5, 5.0 and 10.0 ppm. Final test data were used to generate a 96-hour LC50 of 1.77 ppm by the Binomial Method.

Results from this test were used to determine the treatment levels for the definitive test.

7 of 12 CUl 490 Point =iee=e Road ~ F'er Irg'.cr Ne~ .e'-;ev '--"-. '

c" ' '-.:=c-'"

"e""=~e <<~0~) '""-~ "-'< :r. x

"-.0 Bcx 83C59 ~ -"c c~ Acuge Lc is.ara

RQUR SURVCV, INC.

The definitive test was of 168 hours0.00194 days <br />0.0467 hours <br />2.777778e-4 weeks <br />6.3924e-5 months <br /> (7 days) duration and was conducted using eight (8) replicates of five organisms at each of 5 treatment levels, 0.31, 0.63, 1.25, 2.5 and 5.0 ppm.

Test vessels were examined at each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> interval from test initiation through test termination.

Survival/mortality determinations were recorded for each chamber. Dead animals were removed and discarded.

Chemical A al ses Concentrations were validated at TO, T23, T73, T95, T145 and T168 using Clam-Trol, CT-1 methyl orange method (supplied by Betz Lab, see appendix, page C-1) in the ranges of 0.2 1.0 and 0 ' 3 ' mg/l. A Bausch & Lomb Spectronic 21 was used for the absorbance readings.

Ph sical Chemical Parameters The temperature, pH, dissolved oxygen (DO) concentration and salinity were measured and recorded for each treatment level at the beginning and end of each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> test interval.

Survival Fecundit and Dr Wei ht Upon test termination final live counts were made and all surviving organisms were examined by replicate to determine sex and, with respect to females, egg production. The organisms were then transferred according to replicate to tared weighing boats and oven dried at 105' for 6 hours6.944444e-5 days <br />0.00167 hours <br />9.920635e-6 weeks <br />2.283e-6 months <br />. The weighing boats were then removed to a desiccator to cool ~ Dry weights were determined to the nearest micron.

Survival data for each treatment level was analyzed for lethal toxic effects.

Dry weight data was analyzed for sub-lethal effects on organism growth.

Egg production per surviving female at each treatment level was analyzed for sub-lethal effects on organism fecundity.

8 of 12 499 Point Breeze Pocc,' t:e'er,rgton, stew .'e;sey r 8~ 2 ~ r.',e;:"= 'e ",oQ8)788 9 QQ PQ Box83Q~9 ~ Bc'or 9".Uge ~.o~tsiono 7QGGd ~ re,e,<.-,, '.QQlg 4, o< ( '. ',. -.,

RQUFI SURVCY, INC.

IV. Results The definitive test resulted in total mortality after 168 hours0.00194 days <br />0.0467 hours <br />2.777778e-4 weeks <br />6.3924e-5 months <br /> at the 2.5 and 5.0 ppm treatment levels.

No significant effects on organism survival, fecundity or growth were observed in any treatment level below the LOEC for survival.

Mortality, dry weight and fecundity data are presented in Table I.

The test solution temperature was maintained at 26 +/- 0.5 C (See Table II).

Dissolved oxygen levels ranged between 2.6 and 6.3 mg/l (See Table II). On day 6 the D.O. levels were low (2.6 mg/l); therefore, all test vessels were aerated.

The range and median values of the pH, DO, temperature and salinity are presented in Table II.

The mean concentration validation before each renewal period at TO, T73, T145 and at the end of each renewal period at T23, T95 and T168 can be seen in Table III.

V. Source of Documentation All original data documentation is being maintained at Aqua Survey, Inc., 499 Point Breeze Road, Flemington, NJ 08822, in Betz Laboratories Notebook 002, pages 127 145.

9 of 12 "00 &vier Breeze poco ."er ivglon, Yei~ .'e".e'8Si2

~ ~ "-"-"-"-"-- - (. <<) =~'"-'~>

cx. 05> ~ Bc cr; i'cuge cbisicnc .Ci~-' ~'" ~ "C 06

FIQUFI SURVEY, INC.

Table I:

Summar Sheet For M sid Shrim Survival and Growth Test SURVIVAL DAT Test Conc. Percent of Mean Conc m Re licate Survival Survival 1 0.0 100 80 100 80 80 100 100 100 92 '

2 0.31 100 80 80 100 60 100 100 100 90.0 3 0.63 100 100 100 100 80 100 80 60 90.0 4 1.25 100 100 80 100 100 80 100 100 95.0 5 2.5 0 0 0 0 0 0 0 0 0.0 6 5.0 0 0 0 0 0 0 0 0 0.0 GROWTH DAT Test Conc. Average Dry Weight (mg) Mean Conc m In Re licate Ex osures Wt. m 1 0.0 0.469 0.594 0.296 0.458 0.469 0. 491 0.539 0.401 0. 465 2 0.31 0.485 0.466 0.404 0.477 0.513 0.477 0.418 0.412 0.457 3 0.63 0.431 0.469 0.482 0 '77 0.402 0.547 0.563 0.473 0.481 4 "

1.25 0.506 0.363 0.551 0.402 0.551 0.491 0.422 0.419 0.463 5 2.50 6 5.00 FECUNDITY DATA Test Conc. Fecundity (Percent) Mean Conc m I Re licate Ex osure Percent 1 0.0 100 100 67 100 100 100 67 0 79.3 2 0.31 50 50 100 67 100 100 100 100 95.9 3 0.63 100 75 100 75 100 100 100 100 93.8 4 1.25 67 50 50 100 100 67 100 76.3 5 2.50 6 5.00 10 of 12 4-"99 ~oint Breeze Road ~ FJerning'.cn Yew Je~:.ey G6622 ~ Teieon ne (908) 766 67C PO Scx 6 G59 ~ Ba~cn Rouge '.cubi'iona .'CSS-" ~ Vie"-"c"e 'SGG} 66'-~oS-'~ ~ '"i'" '-

RECUR SURVCV, INC.

Table II: Summar of Ph sical Chemical Parameters Conc. Tem erature H Dissolved 0> Salinit ppm 0.0 (mean) 26. 1 7.8 4.9 28.9 (range) 25.5-26.5 7 '-8.1 2.9-6.4 28.0-30.0

0. 31 (mean) 26.2 7.8 4.9 29.0 (range) 25.5-26.5 7 '-8.2 2.6-6.4 28.5-30.0

0. 63 (mean) 26.2 7.8 4.9 29.1 (range) 25.5-27.0 7.4-8.2 2.8-6.4 28.5-30.0

l. 25 (mean) 26.2 7.9 5.0 30.3 (range) 25.5-27.0 7.4-8.2 2.9-6.4 28.5-30.0 2.50 (mean) 26.3 7.7 4.8 29.3 (range) 26.0-26.5 7.5-8.0 2 '-6.3 29 '-30.0 5.00 (mean) 26.3 7.8 4.9 29.3 (range) 26.0-26.5 7.5-8.1 3.7-6.4 29.0-29.5 11 of 12

~>9 Point Breeze

~

z Rcac ~ F<<evingrcn New,'e<<!e t '8 2 ~ =. e:,".i~', - 7'28), Cc.'"="QC PO Bcx 3050 ~ 8aicn rxcuge L Utstcnc <<'<88' ';=.'e"- = "=,-~q,~,-"- ~

RECUR SURVCY, INC.

Table III Concentration Validation Before Renewal Conc. Mean ppm TO T73 T145 mqm 1 0.0 0 0 ~ 31 0.301 0.278 0.356 0.312 0.63 0.561 0.549 0.523 0 '44 1.25 1.170 1.349 1.150 1.223 2.5 2.330 2.330 5.0 4.220 4.220 Concentration Validation After Renewal Conc. Mean ppm T23 T95 T168 m~1 0.0 0 0 ~ 31 0. 218 0. 152 0 0.123

0. 63 0. 195 0. 157 0.172 0.175

l. 25 0.348 0. 331 0 0.226 2.50 0.831 5.0 2.409 Summary of Chronic Test Endpoints NOEC LOEC **168-HR LC50 1.25 ppm 2.50 ppm 1.37 ppm

~* determined by the Moving Average Method 12 of 12 499 Point Bree:e RoaC ~ Flemington Nev,'e~~ey 08822 ~ 'e;eo+".re,'908) 788."7CC PO Box83059 ~ Ba'on Rouge Lait iona 7C884 ~ '-.-.~e - ~o. '". < >654.4c8-' ~

A N

D I

RECUR SURVCY, INC.

ASI MOC 9100 ~ 04 August 1, 1991 PROTOCOL CHRONIC TOXICITY BIOASSAY FOR MYSID SHRIMP SPONSOR: Bet La o or es c 6 6 So to revose 90 7 STUDY NUMBER: 9 -089 TESTING FACILITY: Aqua Survey, Inc. (ASI) 499 Point Breeze Road Flemington, New Jersey 08822 United States of America PROPOSED START DATE: Jul 6 199 PROPOSED COMPLETION DATE: 99 STUDY DIRECTOR rC'

/ Signature ate QA OFFICER: Rlb g/

Sx t ate SPONSOR APPROVAL:

Signature Date 1 of 7 499 Point'Breeze Poco ~ Flemlngton Ve.v .;-:!.'.ev: 88" ~:ceo~;.,"~ ".08)788'97i 0 .".~Xt "CS) 88"." 4:

PO. Box 8'059 ~ Soton Rouge Louisio.",- '"088-' =-.e ."':;e ',<:-.-~:5~-aj8 'AX.=Cc~ '>"8-'837

RQUFI SURVEY, INC.

PURPOSE:

To estimate the safe or no effect concentration of the test article, using 7-day old mysid shrimp in a seven-day, static-renewal test.

TEST ARTICLES The test article is which was received from Betz Labs c. on 3 the test article is clear li id (description). The sample is labeled with an ASI number -08 Any test article remaining after the test is completed will be disposed of according to instructions from the sponsor or returned to the sponsor.

TEST ARTICLE CHARACTERIZATION:

Data concerning stability, uniformity, composition and additional chemical/physical characteristics of the test article will be provided by the sponsor on the Test Substance Form (supplied by ASI). Chemical analyses will be the responsibility of the sponsor unless prior arrangements have been made for ASI to assume that responsibility.

CONTROL ARTICLES None required.

TEST SYSTEMS h

representing an estuarine/marine invertebrate.

JUSTIFICATION!

Mysid shrimp are considered the appropriate test system as they are a characteristic system in which the response to'toxicological agents can be evaluated.

DILUTION NATERt Whenever possible, natural sea water will be used. If use of natural seawater is not possible, an alternative dilution water, uncontaminated and of constant, quality, will be used.

.langton, 2 of 7 499 Pair t Breeze Road . F!er t rew .'ersey 882" ~ 'e!eon"r e '9C8) 786. 87CG FAX "iG8~ 786 ~lG-"

PG. 3ox 83059 ~ Barcn Rcuge Lcr;isrcna 70864 ~ re!canc.e r8( Q) 5-d-d584 ".AA,50') 92.".8 9

RECUR SURVEY INC SOLVENTS!

Whenever possible, the toxicant will be introduced into the test solution without the use of solvents other than water. If alternative solvents are necessary, they will be used sparingly, not to exceed 0.5 ml/1 in a solution.

TEST MATERIAL!

The technical grade, end-use product or both will be used in this study.

The brand name or chemical names and a lot number are used to define a test material. Mixtures are defined by ingredient and percent of each ingredient, where possible.

TEST ORGANISMS' hl test. Seven (7) day old juveniles will be used to initiate the test.

The age of the organisms will be ascertained from the supplier along with any disease treatment schedule.

EXPOSURE CHAMBER:

Four hundred (400) ml glass beakers or other appropriate chambers will be used.

RANGE-PINDZNG TEST!

If the toxicity of the test substance is not already finding test will be performed to determine the known, a range range of concentrations to be used in the definitive test.

DEPZNZTIVE TESTs A minimum of 40 organisms per concentration will be exposed to five or more concentrations of the chemical. Each designated treatment group will be exposed to a concentration of toxicant that is at least 504 of the next highest concentration.

Test organisms will be divided into eight chambers per treatment group (5 organisms each). All organisms will be randomly assigned to test vessels.

3 of 7

~99 Po!nr Breeze RocC ~ F!er..inc ton ~lew .er.ev 83"."= ~ 'e!e="".ne:Qr 8)138-B C' =',r '9

'RG. Box 3'0"9 ~ Baton Rouge l.osis(cnc 7r 084 ~ reteo =ne ',BGG) o6-'- '66-'AX::-.~; .-'=="-'-"

RECUR SURVCY, INC.

TEST PROCEDURES!

In preparation for the test, the chambers are filled with appropriate volumes of test solution. The test is started by randomly introducing organisms acclimated in accordance with the test design into the test chambers within 1 hour1.157407e-5 days <br />2.777778e-4 hours <br />1.653439e-6 weeks <br />3.805e-7 months <br /> after the addition of the test solution.

Organisms in the test chambers are observed periodically during the test, the dead organisms removed and the findings recorded. Dissolved oxygen concentration, pH, temperature, and other water quality characteristics are measured at specified intervals in test chambers.

~t The g~

test is

~~

The test solution is renewed daily. The organisms are fed newly-hatched daily.

terminated after seven days of exposure. At. termination, dead organisms are removed and surviving organisms in each test chamber are examined for eggs and the sexes determined. Organisms are then prepared for drying and weighing.

ACCLIMATION+

When possible, organisms will be hatched in dilution water. If this is not possible the organisms will be acclimated to the dilution water over as long a period of time as possible. The organisms will be observed prior to testing for signs of disease, stress, physical damage and mortality. Injured, dead and abnormal individuals will be discarded.

Organisms will not be used if if they appear to be diseased or stressed or more than 34 die during the 48 hours5.555556e-4 days <br />0.0133 hours <br />7.936508e-5 weeks <br />1.8264e-5 months <br /> immediately prior to testing.

T ST P E 8 CONCENTRATION MEASUREMENT:

Analytical measurements of the test substance will be made for each concentration at T, T>, T, T<, T><>, and T,< by a method validated before beginning tEe test Ry appropriate laboratory practices.

TEMPERATURE<

The test temperature is 26 + 0.5 C and will be measured at the end of each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> exposure period in one test chamber at each test concentration and in the control.

pm:

The pH will be measured at the end of each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> exposure period in one test chamber at each test concentration and .in the control.

4 of 7

-'99 ~carr 3ree.e Roc< ~ -.ie~ nc'cn )re~"

. RO. 3cx ""C5V Sc.cn:ccure ~cursrcaa

~ /,g4 Kg ~ ~ .

• RQUH SURVCY, INC.

SALINITY'he test salinity will be between 20 and 30 ppt. The salinity should vary by no more than + 2 ppt among the chambers on a given day and vill e measured at the end of each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> exposure period in one test chamber at each test concentration and in the control.

DISSOLVED OXYGEN'he dissolved oxygen concentration is measured at the beginning and end of each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period in one test chamber at each test concentration and in the control.

LIVE COUNTS The number of live organisms in each test chamber are recorded daily, and the dead organisms are discarded.

LOADING<

The size of the test vessels will be such that the loading factor is no greater than one (1) organism per 40 ml of test solution.

PHOTOPERIODC 16 hour light and 8 hour dark photoperiod with a 15 to 30 minute transition period will be employed.

TEST SOLUTION RENEWAL:

The test solutions are renewed daily using freshl'y prepared solution, immediately after cleaning the test chambers. For test solution renewal, the water level in each chamber is lowered to a depth of 7 to 10 mm, which leaves 15 to 20% of the test solution. Nev test solution should be added slowly by pouring down the side of the chamber to avoid excessive turbulence and possible injury to the organism.

AERATION!

Aeration may affect the toxicity of the test solution and vill be used only as a last resort to maintain a satisfactory dissolved oxygen concentration.

5 of 7 "99 P"irt .reeze Rocc

~ ~

~ ."e'er,!rc'.". New .ersey

~ ~

86"." ~ 'e!ec".'re ',9C8; 13c.310 '-;:i.'"-:. ". 3c."".

RG. Box 83C59 ~ 3cicn Rcuge c!::s;cnc

RECUR SuRvev, )WC.

PBBDINQt The organisms are fed newly hatched (less than 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> old) ~litem a once a day from Day 0 through Day 6. The organisms are not fed on Day 7.

PECUNDITYg DRYINQ AND %EIQHINQc At termination, each test chamber is counted, the organisms examined for eggs, the sexes determined and the organisms immediately prepared for drying and weighing. The group of organisms from each test chamber is transferred to a tared weighing boat and dried at 60'C for 24 hours2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> or at 1054C for a minimum of 6 hours6.944444e-5 days <br />0.00167 hours <br />9.920635e-6 weeks <br />2.283e-6 months <br />. The weight of the group of organisms will be recorded to the nearest micron.

ACCEPTABILITY OP TEST RESULTS'he test is acceptable if:

(1) The average survival of control organisms is equal to or exceeds 804; (2) The average weight of control mysids is equal to or greater than 0.20 mg; and, (3) If fecundity used as a criterion in the control is adequate, fecundity should of effect in addition to survival and be growth.

EPPECTS MEASURED!

The end points of toxicity are based on the adverse effects in survival, fecundity and growth. Point estimates, such as LC1, LC5, LC10 and LC50 are calculated using probit analysis. LOEC and NOEC values, for survival, fecundity and growth, are obtained using a hypothesis test approach such as Dunnett's Procedure or Steel's Many-One Rank Test.

REPORT!

The following are included in the final report:

Test Material Xf technical grade product is tested, the source, batch and exact purity are reported. Nhen end-use product is tested, the exact percent of active ingredient and the type of formulation (i.e. granular, wettable powder) are reported.

2 ~ Dilution Mater Dilution water source, its chemical/physical characteristics, and pretreatment are reported.

6 of 7 499 Par'.r 9ree-<<= Rocc ~ ."e'er rngton .".ew.'cree'! 3882 .<ec."o e '~C8 ':~ '"7r "r".:.

PO. ox 8-0=9 ~ ScrcA IccJ'e 'oursror.o;C'88d e:Pie

~

lor a ','-C ) 05~- AX .4.

..'5C'"..:

r < 3'. 88

"-'"" '339

RECUR SURVEY, INC.

3. Exposure Chambers - The volume, diluent depth, and container construction material are reported.

4. Test Organisms Source, quarantine, holding, and acclimatization, inclusive of feeding schedules and disease treatment procedures are reported. The age and scientific names are reported.

5. Range Finding Test - Procedures and results are reported for the range finding study. Included at a minimum are sample size, concentrations tested and mortality data.

6~ Definitive Test - Procedures used to prepare toxicant stock solutions are described. The method of dosing is criteria for determining effects are defined. Raw reported. data or The percentage of deaths/effects at each treatment level along with the number of organisms exposed at each level is reported. Toxic symptoms, both physical and behavioral, observed during the test are reported.

I 7~ Temperature/DO/pH/salinity - Temperature, dissolved oxygen, pH and salinity measurements are reported along with the range and mean of temperature.

8~ Chemical Analyses When chemical analyses are made to validate toxicant concentrations, the analytical method is reported as well as the results. Residue observed in exposure chambers is described in the report.

9. Testing Protocols - This protocol is referenced in the report.

10. Deviations - Deviations from protocol or standard operating procedures are reported.

11. Certification The Study Director and the QA Manager certify the final report.

RECORDS!

All raw data. data manipulations, draft reports and final reports will be retained by Aqua Survey, Inc. for a period of five years from the date of completion of testing. Records will be surrendered to the sponsor upon their written request.

QUALITY ASSURANCE This study will be conducted in accordance with the EPA Good Laboratory Practice Regulations (Federal Register, Volume 48, November 29, 1983) and Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Water to marine and Estuarine organisms EPA/600/4-87/028 May 1988.

7 of 7 49"-. "-~'"r ~r=~-~ Qo c ~ er t~."t~", ~;ev PG.;ox 8" 0="". Bc-cn Acuge !.c~.'tens

~

A P

P N

D I

8

TOTAL ANALYTICALSERVICES FOR A SAFE ENVIRONMENT I

es envronmen a,.

Project No.t 9117891 Log in No. t 7861 P.O. No. t Pending Oate )April 30, 1991 ANALYTICAL OATA REPORT PACKAGE FOR Aqua Survey lnc.

499 Point Breeze Rd.

Flemington, HY 08822 ATTNt Jim Todd REFt Lab Hater SAMPLE LABORATORY SAMPLE IOENT IF I CATION NUMBER HAIR IX SEE HEXT PAGE

)IE CERTIFY THAT THiS REPORT IS A RESPE LLY SUBMIT EO, TRUE REPORT OF RESULTS OBTAINE0 N ENV ONMENT L INC FROI OUR TESTS OF THIS HATERIAL.

PARAQ K. SHAH, Ph 0.

ORGANI C LAB~ MANAGER OOI NEELEY ORY OIRECT R

/J art¹ 73469

))aport on samptefs) furnished by ct)ent appttee to swnpiels). Report on sarnpiels) obtained by us applies only to tot samptetL lnfonnaoon contwned helen is not to be used for reproduction except by spec>>t perm>>won. Sample(s) ~II be retwned lor thiny days maximum after cate ol repon unless specificaay reouested othenese by dient In the event that there are ponions or pats ol sampk(s) remwning atter Nytest has completed the rethured tests. Nytest shat) have the option ol retumetg such sample(s) lo the ct>>nt al the ctienl's ex pense.

box 153 8 a 60 seaview blvdport washington, ny I 3 050 a {5<6) 625-5500

N EST RO C~

LABORATORY SAMPLE TYPE OF NUMBER IDENTIFICATION SAMPLE 7861001 LG SALT Water 7861002 LG FRESH Water 7861003 LP FRESH Water

nytest environment@ .

DATA REPORTING QUALIFI "RS U Indicates compound was analyzed for but not detected. Report the minimum detection limit for the sample with the U (e.g. 10 U ) based on necessary concentration dilution act'ons. (This is not necessarily the instrument detection limit.) The footnote should read U-Compound was analyzed for but not detected. The number is the minimum attainable detected l'm't for the sample.

Indicates an estimated va'ue. This flag is'sed e'ther when estimating a concentration or tentatively identif compounds where a 1:1 response is assumed o when the mass

'd spectral data ind'cates "he presence of a co-pound that meets the identification criteria but the result 's less than the specified detection limi- but greater than zero (e.g.: If limit of detection is 10 ug/'nd a concent"ation of 3 ug/l 's calculated, report as 3 J. )

Th's flag is used wnen t"..e a.".alyte is found in the blan~

as well as a sample. It '.".dicates possible/probable blank contaminat'on ana warns ='.-. d =a use= to take appropr'e action.

T This flag ident f'es a'a""o-ed compounds '.".at were found above the method detec"'.-. '.='=s.

NA Th's flag indicates tha= =.-.e =ompounds are noi appl'c ble; A Aldol condensation produc".

Note: Data on soil samp'es ev-ressed on a dry we'ght basis.

All non-water samples a o "eported on soil m'scellaneous.

forms. Th's includes samples whose matrix is listed as The initial and cont'nuingl's"ed calibration dates and t'mes for the volatile fraction are on the BFB summary forms. The initial and continuing calibrat'on dates and times for the are listed on the DFTPP summary semi-volatile fractions forms'0003

nytest environments Report of Tests Water Suitability Test We find as follows:

The sample was analyzed according to Standard Methods for Examination of Water and Wastewater, 14th Ed., page 888.

Growth Ratio Sample ZD: LP FRESH F 08 Lab XD No.: 7861003 Remarks:

The sample is of acceptable suitability for the use 'n bacteriological testing. A ratio greater than 3.0 would indicate growth promoting substances present in the water. A ratio less than 0.8 indicates the presence of growth inhibiting substances.

OO000

nytest environmental REPORT OF ANALYSZS Log Zn No.: 7861 We find as follows:

Parameter(s) Sample Zdentif ication LG SALT LG FRESH LP FRESH Method (7861001) (7861002) (7861003) Blank pH 7. 66 NA Salinity, ppt 29 ' <1 Results in mg/l:

Ammonia Nitrogen 0. 130 0 22 <0.05 Bromide 40 ' <1.0 Fluoride 0.576 <0.02 Zodine <1 <1 Nitrate <0.04 <0.04 Phosphate <0.04 <0.04 Residual Chlorine <0.1 <0.1 <0.1 <0.1 Sul ide <0.10 <0.10 Sulfate 1910 <1 Total Cyanide <0.01 <0.01 Total Suspended Solids 3 <1 ppt ~ parts per thousand 00005

nytest environments,.

REPORT OF ANALYSIS Log Zn No.:7861 We find as follows:

Results in mg/1:

Parameter(s) Sample Identification WWW LG SALT LG FRESH LP FRESH Method (7861001) (7861002) (7861003) Blank Aluminum <0.2 <0 '

Arsenic <0.010 '<0.010 <0.010 Beryllium <0.005 <0.005 Boron 3.3 <0.1 Cadmium <0.005 <0.005 <0.005 Calcium 271 <1.0 Chromium <0.010 <0.010 Cobalt <0.05 <0.05 Copper <0.025 <0.025 <0.025 I on <0.050 <0.050 Lead <0.050 <0.050 <0.050 Magnesium 932 <1.0 Manganese <0.015 <0.015 Mercury <0.0002 <0.0002 <0.0002 Molybenum 0. 16 <0.010 Nickel <0.040 <0.040 <0.040 Potassium 348 <1.0 Selenium <0.010 <0.010 Silver <0.010 <0.010 Zinc <0.020 <0.020 <0.020 Total Organic Carbon 20.5 11.3 <1.0 00006

10T NYTEST ENViRONMENTAL lNC.

TCL PESTlCIOE/PCS ORGANlCS ANALYS!S OATA SHEET SAMPLE MATRlXt MATER SAMPLE lot LG SALT CONC. LEVEL: LOM LAS SAMPLE lot 7861001 EXTRACT10N OATEt 4/9/91 01L FACTOR: 1.00 ANALYSlS GATE: 4/18/91 X MolSTUREt NA UG/L CMPO ¹ CAS Nunber PESTlClOE/PCS CCSlPOlNO I 319 84.6 I Alpha BHC 0.050 U.

2 I 319.85-7 l Beta-BHC o.oso u.

3 l 319-86-8 l Oelta-BHC o.oso u.

4 l 58 89-9 l Ganna. 8HC(Lindane) o.oso u.

sl 76-44-8 l Heptachlor o.oso u.

309.00.2 l Aldrin 0.050 U.

7 l 1024-57-3 l Neptachlor Epoxide o.oso u.

Sl 959.98-8 l Endosulfan l o.oso u.

60-57. 1 l Oieldrin 0.100 U~ I 1o l 72.55.9 4,4'OOE 0.100 U. l 70.20.8 l Endrin 0.100 U. I 12 l 33213.65.9 l Endosulfan lt o.loo u. l 13 I 72.54-8 l 4,c.ooo 0.100 U. I 1031.07.8 l Endosul fan Sulfate 0.100 U I 15 I 50.29.3 l 4,4'-ooT 0.100 U. I 16 I 53494-70.5 l Endrin Ketone 0.100 U. I 17 I 72-43-5 l Methoxychlor o.soo u.

18 I 57-74-9 l Chlordane o.soo u.

19 I 8001-35.2 l Toxaphene F 000 U. I zo l 12674-11-2 l Aroclor-1016 o.soo u.

21 l 1110C.28.2 l Aroclor-1221 o.soo u.

22 I 11141-16-5 l Aroclor-1232 o.soo u.

23 l 53469.21.9 l Aroclor-1242 o.soo u.

24 l 1267Z-29.6 l Aroclor-12CS o.soo u.

25 I 11097.69.1 l Aroclor-1254 1.0oo u.

11096 82.5 l Aroclor-'l260 1.000 U. I I l D-7 00007

ORGANOPHOSPHOROUS PESTICZDES Log in No.: 7861 Sample ZD No.: LG SALT Lab ZD No.: 7861001 Detection Limit Parameters (ug/1) (ug/1) Found Vapona (Dichlorvos) 0.2 ate

!'hor 0' ND Diazinon 0.6 Naled (Dibron) 0.2 Dementon - S 0.2 Disulfoton 0' Ronnel 0.3 ND Chlorpyrifos (Dursban) 0.3 ND Bolstar (Sulprofos) 0.2 ND Stirophos 2.0 ND Fensulfothion 5.0 Azinphos methyl (Guthion) 5 Coumaphos 5 Malathion 0.3 Parathion 0.3 ND ND ~ Not Detected

'1 0-T XYTEST EXVLROXXEXTAL lXC.

TCL PFSTlClDE/PCB ORGAXlCS AXALYSlS DATA SHEET SAMPLE NATRlX: MATER SAXPLE ID: PBLK1 COXC. LEVEL: LOM LAB SAXPLE lo: A040005 EXTRACTLOX DATE: 4/9/91 0'iL FACTOR: 1.00 AXALYSls DATE: 4/19/91 I XOlSTURE: NA UG/L CAPO ¹ CAS Xunber PEST lCIOE/PCB CQIPOUXO 319 84.6 I Alpha BHC 0.050 U.

2 I 319.85-7 ( Beta-BHC 0.050 U.

3( 319.86-8 ( Delta BHC 0.050 U~

58-89-9 Gamna-BHC(Lindane) 0.050 U.

<<( 76-44-8 ( Heptachlor 0.050 U.

6( 309 00-2 ( Aldrin 0.050 U.

7 ( 1024.57.3 ( Heptachlor Epoxide 0.050 U.

8 I 959-98-8 ( Endosulfan l 0.050 U.

9 (

60.57-1 (

Dieldrin 0.100 U.

10 ( 7Z-55-9 (

4,4'OOE 0.100 U.

11 I 70-ZO-S I Endrin 0.100 U.

1Z ( 33213 65 9 Endosulfan 1 I O.10O U.

13 I 72-54.8 C,C-OOO 0.100 U.

14 I 103'1 8 ( Endosulfan Sulfate 0.100 U.

15 I 50-29.3 (

C,4'OOT 0.100 U.

16 53494-70 5 ( Endrin Ketone 0.100 U.

17 I 72-43-5 ( Xethoxychlor 0.500 U.

18 I 57-74-9 (

Chlordane 0.500 U.

19 (

8001-35.2 ( Toxaphene 1.000 U.

20 (

1267C-11. 2 (

Aroclor-1016 0.500 U.

21 (

11104-28 2 (

Aroclor-1221 0.500 U.

22 I 11141.16-5 ( Aroclor.1232 0.500 U.

23 (

53C69-21 9 (

Aroclor -1242 0.500 U.

24 (

12672-29.6 ( Aroclor.1Z48 0.500 U.

25 I 11097-69. 1 (

Aroclor.1254 1.000 U.

26 I 11096-82.5 (

Aroclor.1260 1.000 U.

I D-9 Ooo>1

ATTACHMENT8 A - Letter of 7-19-91 from Ms. Joan M. Karnauskas, Chief, Permits Section, US EPA Region V, to Mr. Larry Lyons.

B - Letter of 8-20-91 from Dr. Donald S. Cherry, Virginia Polytechnic Institute and State University, to Mr. Larry Lyons.

C - Letter of 9-20-91 from Dr. Dwight P. Davis and Mr. Larry A. Lyons, Bezt Laboratories, Inc. to Ms. Joan M. Karnauskas and Mr. Peter Howe.

4 4 July 19, '.991 5WQP-TUB-8 Mr. Larry Lyons Office of Water Management Indiana Department of Environmental Management 105 South Meridian Street Indianapolis, Indiana 46206-6015

Dear Mr. Lyons:

Region V reviewed Betz Laboratories, .Inc. (Betz) June 6, 1991, letter and attachments related to its Clam-Trol CT-1 (CT-1).

Region V does not concur with the'etz position as stated in the letter. Our concern is that all of the necessary information required to establish appropriate numerical water quality based effluent limitations for this product has not been limited information indicates that, attainment of developed.'urthermore, water quality based effluent limitations based upon most and chronic water quality standards may not be possible as States'cute the product is currently used.

This is due in part to the possibility that the "detoxification<<

process may result in acute toxicity to some organisms. Your general treatment, as we understand, consists of applying CT-1 at 10-15 mg/l as product and detoxification is met by applying bentonite clay at a concentration 3 times higher (3:1 ratio) than that of the product. Recent studies by Cherry et al. (1990)~

documented a Cerioda hnia LC~O for bentonite clay of 16.5 mg/l which is considerably lower than the approximately 30-45, mg/l bentonite clay added in the" detoxification process. The Cherrv et al. (1990) studies also documented high mortality to Cerioda hnia from bentonite clay and CT-1 treated effluent (10 mg/l CT-1 and 30 mg/l bentonite clay) at 54 effluent (Table 3), although control mortality was high at 154. The range finding test in Table 13 also documented 100% mortality in 50 effluent with no control mortality. In addition, the Cherry et al. report documented chronic effects to Cerioda hnia. Finally, acute toxicity testing hy Betz on ~Da hnia macana also found 304 mortality at a CT-1 and bentonite clay concentrations of 10 and 30 mg/l, respectively.

'herry, D.S., J.L. Farris, J.R. Bidwell, A. Mikailoff, R. L.

Shema, J.H. McIntria (1990). 1990 Corbicula Control Program Env'onmental Fate and Effects Studies Baseline- and Spring Do=','. =.'= ":==-. Duquesne Light Company, Beaver Valley Power St<

We are als'o mystified as to why the Cerioda hnia chronic (NOEL and LOEC) estimates were reported in terms of nominal concentrations while the lab data sheet indicates that the measured concentration was approximately one half that of the nominal. Please be advised that Region V cannot accept either acute or chronic data based on nominal concentrations unless measured concentrations are within 804 of the nominal concentration throughout the test. If this is not shown to be the case, LC,~'s must be determined from measured concentrations.

There is another major issue associated with the use of CT-1, namely its fate in sediments. This is of particular concern in anaerobic sediments where the surfactant may not be degraded and/or act as a bacteriostat to anaerob'c organisms. We have discussed this issue with you on a number of occasions, and it has yet to be addressed. It is our understanding that there has been a recent report prepaxed by a committee of the Chemical Manufacturing Association (CMA) directed specifically at quaternary ammonium surfactants. Included in the report are studies of the environmental fate and biodegradation of a number of these cationic surfactants. We would appreciate a copy of this report.

Region V has also recently become aware that it may take 15-30

'minutes for CT-1 to be fully "detoxified" with bentonite clay in some situations. If this is the case, there may be significant releases of active surfactants that can negatively effect the receiving stream. As you are aware, transit time in many once through cooling systems is frequently less than 5 minutes. This may explain why several users of your product reported foaming in the discharge area. Any effluent samples for specific chemical analysis or toxicity. testing subjected to holding times of several hours may effectively disguise potential toxicity in the receiving stream. Unless Betz has conducted studies to document the rate at which detoxification occurs in the presence of bentonite clay, studies will have to be undertaken to address this issue.

Based on these observations, of CT-1 could be accomplished itwithin appears unlikely that discharge the constraints of current state water quality standards. Further, in the absence of environmental fate data, Region V would not concur in the future with use of CT-1 when it is discharged to harbors, bays, lakes, or rivers where anaerobic sediments and habitat degradation already exist, as this would be inconsistent with federal and state antidegradation policies.

Zf Betz would like U.S. EpA to further consider this issue, Region V is requesting that Betz supply us with the following info&ation.

1. Copies of full reports related to all acute and chronic toxicity testing for CT-1 and its detoxified products.

These reports need to document whether acute and chronic toxicity results are based on nominal or measured concentrations, and whether the toxicity tests are flow through, static renewal, or static bioassays. Results of all analytical measurements need to be included.

2. A copy of the CMA report for quaternary ammonia compounds, if it is possible to obtain .the report.

3. Copies of all biodegradation studies of CT-1 surfactants submitted to U.S. EPA as part of the FIFRA registration process. It is our understanding that such studies were conducted by the firms from which Betz purchases the surfactants in CT-1.

4. Results of any studies which documents the rate at which bentonite clay detoxifies CT-1.

5. Betz's intent or plans for additional toxicity and environmental fate studies.

If you have any questions, please call Peter Howe at {312) 886-0233.

Sincerely yours, Joan M. Karnauskas, Chief Permits Section cc: Mr. Bill Jung, Betz Industrial Lonnie Brumfield, IDEM Heidi Nassiri, IDEM Bill Creal, MDNR Brenda Sayles, MDNR Bob Schacht, IEPA Gary Kimble, MPCA Rick Magni, OEPA PHLYONS.LTR

B COLLECE OF ARTS AND ECIENCF re>q~

VIRGINIA POLYTECHNIC INSTITUTE AND STATE UNIVERSITY ji, Blacktburg, Virginia 24061-0406 DEPARTMENT OF BlOLOGY (703) 231W07 August 28, 1991 Mr . Lar ry Lyons Regulatory Aff'airs Section BETZ Laboratories 4656 Somerton Road Trevose, PA 18947

Dear Mr. Lyons:

The purpose interpretation of of'his data from my letter is to clarify the 1998 report to Duquesne Light Company (Corbicula Control Program -'nvironmental Fate and Ef'f'ects Studies Base 1 ine, Spr ing and..Fal 1 Dosing Studies )

as discussed by Ms. Joan M. Karnauskas of the US Environmental Protection Agency. I will discuss these issues point by'oint per her letter of'uly 19, 1991.

Page 1, paragraph 2, lines 6 9: Reference to the 1)

LC58 f'r bentoni te clay of' 6. 5 mg f'r Ceriodaphnia should. be taken in its complete, context relative to the data in Tables 1-5 of'y report. These acute tests, part of a baseline study, were carried. out at a time when the Ohio River water was subject to excessive surface water runof'f during spring seasonal precipitation. As a result, ceriodaphnids were periodically stressed. f'rom the runoff contaminants as is especially evident in Table 2 (control mortality of 55$ time in river water and, 8$

died in 188$ effluent).

2) Page 1, paragraph 2, lines 9-15: As we repeated some of these tests (Table 5) with additional river water, we still had unacceptable control mortality (15K). Ve carried. out these tests gust prior to the spring dosing of the plant f'r hacL no control over the weather or the timing of clam control and the dosing schedule. It was evident to us {my researchers and myself') that the Ceriodaphnia culture acclimated to Ohio River water was substandard or partial ly str essed at this . The cadmium ( Cd) reference toxicity tests carriecL out during these turbid and latter clear river conditions

page two substantiated our concerns (Table 5 ) . Ceriodaphnia were twice as sensitive during the turbid water conditions of the spring dosing period versus two months later after the river had cleared (ie., LC58

8. 87 ver sus 8. 17 mg Cd/L, respectively ) . Even though the river water was filtered prior to acclimation and testing, the f'iltering process removed only solids, not dissolved contaminants.

3) Page 1, paragraph 2, lines 15-16: The comment that the "Cherry et al. report documented chronic ef fects to Ceriodaphnia" needs special clarif'ication.

As determined by US EPA protocol, chronic effects become noteworthy if they occur at the instream waste concentration (IWC) which was calculated at 5g ef'fluent in this case. There was no organism toxicity or reproductive impairment at or near the IWC for Ceriodaphnia in all chronic tests run (Tables 1 4, 1 5, 1 6, 28, 29 and 38 ) . Repr oduc tive impairment occurred at 58 and 188% ef'fluent exposures in the spring and only at 188$ effluent in the fall. Hence, impairment consequences were at least 18 fold or higher relative to the IWC.

Page 1, par agr aph 2, back to l ines 6-9 r egar ding clay toxicity and Ohio River stressed conditions f'r Ceriodaphnia: Since clay at 6. 6 ppm ki lied 58$

of'he ceriodaphnids in 48 hr in the spring 1

baseline tests where Ohio River water was deemed stressful by itself (Table 4), why were the same test organisms able to survive for 7 days and reproduce in 48$ effluent in the fall (Tables 28-29)'P Note that total suspended solids from the clay addition for CT-1 detoxification of the effluent was up to 43.3 ppm (Table 47). By taking 48$ of the TSS from clay in the effluent (43.3 ppm x 8.48 = 19.3 ppm), the TSS level of test exposure in the fall dosing (Tables 28 and 29) with bentonite clay in the effluent exceeded the laboratory LC58 data concentrations generated in the spring. Clearly, the data indicate that Ceriodaphnia under acceptable testing (non-stressed) conditions in the fall of 1998 can withstand turbidity from clay at levels higher than the LC58 obtained in the spring. In addition, the data in Table 29 also refute the concern of bentonite clay toxicity to Ceriodaphnia. The e ffluent tested in the fallofafter one week of aging contained up to 43.3 bentonite clay at 188$ concentration but had.

ppm no significant impact upon Ceriodaphnia survival and.

page three reproduction. In fact, reproduction at lSSW effluent (18.3 neonates) exceeded that found in the controls (15.7).

5) Page 2, paragraph 2: Please note that we carried out sediment toxicity tests before, during and after dosing of the Duquesne plant (Tables 18, 19, 28, 33, 34 and 35). There was no short or long-term toxicity or impairment due to CT-1 and/or clay deposition to the test organism (Chironomus riparius) in any of these tests. We will be glad to address the issue of potential anaerobic sediment consequences with CT-1:DT-1 influence with future research.

Also note in Tables 28 and 29 that the toxicity in 188$ effluent is short term. Impairment to Ceriodaphnia in 188$ ef fluent during plant dosing (Table 28) was observed, but the same ef fluent tested one week later caused no toxic or impairment consequences (Table 29) . Therefore, the physical binding of CT-1 with DT-1 does occur rapidly.

6) Page 2, paragraph 3: The thought that it takes 15-38 minutes for CT-1 to be fully "detoxified" with bentonite clay is inaccurate. The studies by Cherry et al. (1998) at Duquesne Light Company indicated that CT-1 was bound. by clay almost instantaneously or within the first minute of contact time since CT-1 was not measurable in the effluent receiving site gust prior to the effluent sampling point to the river. We believe that this detoxification rate issue can be addressed by a series of acute toxicity tests over a short period of time.

Overall, I am concerned about the problem of "proof-texting", that is, pinpointing data in one table (Table 4) and generally ignoring the theme of the data in Tables 1 through 5 of my report. We concluded that surface water runoff conditions in the spring 1998 baseline studies influenced the fitness of the test organisms at that time, and this consequence should be acknowledged and factored into the overall policy-making process of CT-1 utilization for Asiatic clam control. I also believe in the "weighted data" approach in hazard assessment. We used three tiers of toxicity testing (formal laboratory, artificial stream microcosm and in-river validation studies) with six test organisms ranging from invertebrates to fish. The overall conclusion from this comprehensive testing strategy was that no immediate or long term (ie., 48 days after dosing) toxic consequences to the laboratory test organisms or the endemic biota were evident in the Ohio River at or near the IWC.

page four Please feel free to call necessary or if clarification me if additional information is is required.

Sincer ely, p'onald S. Cherry, Ph.D.

Professor of'oology DSC/sbc cc: J. Bidwell Biology Dept., Va Tech J. Cairns, Jr.

Director, UCEHMS, Va Tech R. Shema Aquatic Systems Corp.

V. McIntire Duquesne Light Co.

FRQti:RLD E.L. ,0: . 48'7 697 6985 OC~ 4 '9': 3?M P. 82 hl 'ABORATORIES, INC.

gQMEATQN ROAO>> TPHVOSE, P< 19053 6783 U,S,A, / TEL: 215

~ 355-3300 ~ PAX ¹ 355.2BBB September 20, 1991 Ms. Joan M. Karnauskas, Chief U.S, EPA/Region 5 Permits Section 230 South Dearborn Street Chicago, IL 60604 Mr. Peter Howe U.S. EPA/Region 5 230 South Dearborn Street Chicago, lL S0604

Dear Madam and Sir:

This letter is responding to each of the questions and inquiries addressed ln your letter of July 29th. First, responses and comments (See A to E) will be provided for those inquiries on pages 1 and 2 followed by the information being requested on page 3.

A letter (Attachment A) prepared by Dr. Donald S. Cherry regarding the environmental Impact study of the Clam-Trol CT-1 applications for Duquesne Ught Company's Beaver Val/ey Nuclear Power Plant discusses the details of the 1990 Spring and Fall applications. This study was designed to determine if the lnstream Waste Concentration (IWC) of the discharge water during and after these treatments would have a negative environmental impact to thIs vicinity of the Ohio River. These studies, employing a tier approach with bloassay organisms and endemic biota, have demonstrated that a 10 to 20 fold increase above the IWC was achieved. Your questions regarding the Ceri studies are addressed by Or. Cherry.

B. It should also be noted, regarding the Asiatic clam treatment program at the Beaver Valley Nuclear Plant, that Duquesne Light Company had thoroughly investigated the available methods of macrofoullng control. Besides the CT-1 treatment approach, only oxidizing biocldes could be considered. It was dearly recognized that daily intermittent chlorine treatment programs have never proven to be effective. Only continuous chldrinatlon treatments for several weeks are able to eradicate the residing clam populations in a cooling system, The 'ffectively application of a continuous chlorination and dechlorination treatment program to a

It'

Q I II

FRY:RLDPE ~ LE 487 69" 6985 OCt 4g 1991 '?:38'.83

+'Lf'K ~oanoaea.wc US EPA, Region 5 Page 2 once-through cooling system presents many operational and safety problems. The CT-1 treatment program for Asiatic clam fouling control usually requires 2 to 3 applications/year depending upon the degree of infestation of the juvenile clams, spawning season, and growth rates. Most treatments require a 12-hour application.

C. The acute bloassay of the clay/CT-1 mixture (30 mg/L to 10 mg/L, respectively) with ~ee>~hni ~ma no evaluated the detoxification potsnilalor the daysinlaboratory culture water. Unlike river and lake water, the laboratory culture water does not contain any additional constituents (ie., suspended solids, hurnlc acid, silt) that would exhibit further demand upon the CT-1 actives. This bioassay showed a 70%

survival in the presence of the clay/GT-1 mixture. During actual GT-1 applications to cooling systems, a total or cumulative demand ls expressed by the adsorption of the CT-1 actives by the constituents of the cooling water and the demand of the mlcrofouled surfaces of the cooling system, In addition to any detoxification by clays that may be applied.

D. Attachments 8 and C Include a letter addressed to Ohio EPA (Feb. 1S91) and a bt yph d thh tdl Tttb~g I.yh glh p<< lt lith pll I I Mdlgth ydy~g'ta hl chronic testing protocol and the implications related to the CT-1 product. In brief, the Ce oda hnia protocol requires feeding the daphnids with an algal Inoculum on a daily basis. If the algal feeding regime ls reduced or altered, a significant impairment in daphnld reproduction results that ultimately Invalidates the chronic bioassay. In addition, when the algal Inoculum is Introduced to the CT-1 test concentrations, a decrease of the measured CT-1 concentration results from the adsorption of the CT-1 actives by the algae.

ThthrLddhlgh tbl ylhltbtg I dlly Ilt mentioned studier This bloassay another bloassay conducted following the above assessed the effect of a 12-hour exposure following the standard 74ay protocol.

bl T,P~GI1/2-day recovery period.

12-hour period followed by a 6 p dt'gytt Test concentrations were measured at 0 hr and 12 hrs. The measured concentrations were the average concentrations between these time intervals. The algal inoculum dkf reduce the initial measured CT-1 test conpentrations to low or nondetectable levels (~0.2 mg/L). Subsequently, this bioassay presented the LOEL's (Lowest effect levels)-

for Cedodaphnia reproduction at 0.8 rng/L and the NOEL (No effect level) at 0.4 mg/L as the nominal test concentrations. The nominal ohronio value was caicuiated to be 0,57 mg/L and the measured chronic value to be 0.28 mg/L The completed report on this study has not been received by us as yet. A data sheet

~ ~

FROM:R8D E.L. TQ'87 697 6985 QCT 4r '99: 7:39AM P 84 g3+'Lf'+

US EPA, Region 5 Page 3 that was sent to us by FAX is being included here as Attachment D. The final report will be sent when received by us.

E. Anaerobic Sediment Study/Calumet Region Three sediment samples from the Calumet River have been taken and sent to Betz'icrobiology Laboratory. A forrnal report has not been issued as yet but they have verbally reported a low level of biological activity, The levels have been determined to lie between 104 and 10~ per ml, When a report ls Issued it w)ll be sent to you.

The information requested on page 3 of your July 29th letter is being presented, as follows:

Attachment h presents the acute toxicity data sets fore'r-t with ~Da hnia

~ma na fathead minnow, bluegill sunfish and rainbow trout. These data sets include the 4B-hour screen tests, the definitive LCM bloassays and the statistical analyses for computing the LCgo s. Ail dsata sets for these static acute bloassays are presented as nominal test concentrations, but data are available with measured concentrations as noted below and In the chronic rlod hnia studies addressed above.

Attachment F presents the 7<ay static renewal chronic bloassay with fathead minnows. CT-1 test concentrations were measured at days 1, 4 and 7. It Is interesting to note for this 7 day exposure with the fathead minnow larvae that the LCg0 determination is greater than 3.2 mg/L This estimated chronic LC~0 value ls higher than the LCq0 value determined from the static 96 hour0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> acute bioassay of 3.0 mg/L. The fathead minnow has shown to be the most sensitive fish species.

2. The Chemical Manufacturing Association (GMA) was contacted to Inquire about a recent report available on surfactants, A report from the "Alkyiphenol

& Ethoxylates Panel" is presented as Attachment G. Please note that these surfactants are not related to the aclives in the Betz Clam-Trol product,

3. Clam-Trol CT-1 is an EPA registered End-Use formulation composed of two EPA registered active ingredients listed below. These registered active ingredients are currently involved in the reregistration process mandatecl by the 1988 FIFRA amendments. The Environmental Fate studies as required by

FRY:RLD E.L. To: 487 697 6985 OCT 4 > 199': 39At1 P. Q5 93DN7(%

US EPA, Region 5 Page 4 40 CFR Part 158.130 are in various stages of development in response to the reregistration process. Wa will summarize to the best of our ability the current status of these active ingredients, however the manufacturers have been reluctant to share information with us until the process has been completed.

Alkyl Dimethyl Senzyi Arnmonlum Chloride (ADBAG)

A consortium af manufacturers formed a data development task force within the Chemical Specialties Manufacturers Association (CSMA). The ADBAC Committee rias been very protective ot tnese oata because oi inc economic value they have for the registration of this active on a worldwide basis. Ne understand that these data have been submitted to the Pesticide Registration Division (GTS) at USEPA and the committee Is awaiting agency review and acceptance prior to the release of these data to other interested parties.

Dodecylguanidine Hydro:hloride (DGH)

The manufacturer of Dodecylguanldine Hydrochloride has submitted hydrolysis, photodegradation and adsorption/desorptlpn studies to the Pesticide Registration Division of the USEPA. The Aeroblo and Anaerobic Aquatic Metabolism studies are in progress and are expected to be completed in the next feI months.

Due to the apparent slow pace of the pesticide registration review resulting from the massive undertaking of the reregistratlon process, and the need for environmental fate information to address questions by EPA and state agencies; Betz has decided to condL 4 a biodegradability study on the Clam-Troi GT-1 product. This study should be completed in a few weeks.

4, The rate of detoxification could essentially be defined as instantaneous. Both of the GT-1 actives are readily adsorbed by a variety of materials and reduced to nondetectable levels. This rapid detoxification process has been verified by previous application experiences. In almost all GT-1 treatment programs to nnnn kahn nnnilnn cuba Nc mhara rfatov Jfio>tihn Lvith chv iS reouited. the s

residence time from the point of clay feed ln the discharge canal to the point of discharge is usually less tpan one minute. Analytical determinations of residual CT-1 at the discharge have consistently confirmed the rapid detoxmcation of the CT-1 actives. The treatment program at the Beaver Valley Nuclear Plant discussed by Dr. Cherry (Attachment A) also confirms this rate of detoxification. Other case histories where the rate of detoxification has been analytically verified include the Cook Nuclear Power Plant on Lake

t~ j S ~

I I p '>>

FROrl:RED E.L. to: 487 69? 6985 '?:aBM P.86 lXtX'LI'K US FPA, Region 5 Page 5 Michigan; 9-Mile Point Nuclear Power Plant on Lake Ontario; and a Good Year Tire Plant, Gadsden, AL.

5. Updates and plans for additional studies are as follows:

This past season (June to October) on-going studies in two bioassay trailers situated at the J. R, Whiting Plant on Lake Erie have been evaluating various chemical treatment strategies for fouling control of zebra mussels. One of the objectives of this study was to evaluate the effectiveness of dally intermittent chlorine and bromine applications. The performance of the treatment programs Is being determined by the rate of larval setting, the accumulation of juvenile (post laival) stages, mortality, and growth. The intent of the evaluation is to determine the merit of these fouling control strategies, in order to make appropriate treatment recommendations. However, the preliminary results are demonstrating that the daily 2-hr/day Total Residual Oxidant {TRO) treatments have vanable to limited degrees of effectiveness In preventing the setting of the larval stages. One other relevant observation that is being assessed during this study is the significant entrainment and settlement by the post.

larval stages of the zebra mussel, These post-larval stages are able to pass through screens and settle within cooling systems and are particularly resistant to the intermittent TAO applications. The growth rates of Juvenile and adult mussels are also being determined from 30 to SO days of application. As has been reported by other investigators, the growth rate is not being Impacted by dally intermittent applications.

Other experiments are being conducted during this study to evaluate the minimum treatment dosages for CT-1 applications and the number of applications that will be required to achieve various degrees of fouling control.

seteotsd tax{atty {Lean) btoassays {t.e, ~Da hnta ~ma na fathead mtnnow) will be rerun to compare tFe LC<n determinations between measured and nominal test concentrations. These KC50 determinations will be made from 96-hour exposures as well as 12-hour exposures that more accurately model actual treatment programs. These Initial bioassays will be completed by October 30th.

LC50 determinations to Zebra mussels and Asiatic clams under flow.through test conditions have been completed (See Attachment H). Note that acute (72-hour exposure), intermediate (144-hr exposure) and chronic (15 day exposure) LC50 s are determined also, and that the acute/chronic ratio ls less than two.

h ls

'h P

~

FR&1:RED E.L. "0: 487 697 6985 OCT 4s '99': 4'll P. 0"

~aQ'~

US EPA, Region 6 Page 6 Anaerobic biodegradation studies are being planned. Attachment I is a letter drafted by the Betz microbiology group to initiate the negotiation of an anaerobic testing protocol as there is not one that Is generally or widely accepted. It is Betz'ntent to conduct an anaerobic study once a protocol is adopted.

We believe that this letter addresses all the Issues raised by your 29 July 1SS1 letter. We also feel that there are sufficient acute and chronic data to allow a numerical water quality standard to be set for Betz Clam-Trol GT.1.

Since there appears to have been some misunderstanding or miscommunication of, study results, we would like to request a meeting with you to discuss the issues contained in the 29 July letter and this response plus any other associated tssues. After you have had an opportunity to review this letter, we will call to set up a meeting date and time.

Very truly yours, BORATORIES, INC.

D wight P. Davis, Ph.D.

Assistant Vice President Environmental Affairs f'7

~p~.~l Lany A. Lyons Manager, Aquatics Group DPD/LAL cn CC: Lonnle Brumfield, IDEM Heidi Nasslrl, IDEM Bill Creal, MDNFI Brenda Sayles, MONR Bob Schacht, IEPA Gary Kimble, MPCA Rick Magni, OEPA Bill Jung, Betz Industrial Division Ray Matuza, Betz MetChem Division

E

'0