Text

Detoxification of Clam-Trol CT-4 & CT-1:Daphnia Magna Bioassays
	ML20210J873
Person / Time
Site:	Perry
Issue date:	04/07/1994
From:	Evans K, Lyons L; BETZ LABORATORIES, INC.
To:	;
Shared Package
ML20210J829	List: ML20210J824; ML20210J873;
References
	NUDOCS 9708190082
	Download: ML20210J873 (9)
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.sa e. ssin PY-CE1/0 EPA-0275L Page 1 of 9 4

DETOXIFICATION OF CLAM TROL CT 4 AND

~

CT 1: DAPRNIA MAGNA BIOASSAYS

\\

4 Study Coriductb for:

Great Lakes Steel

.I Study Conducted by:

Bet Laboratories,Inc Aquatie Toxicology Laboratory One Quality Way Trevose, PA 19053 a

Larry A. Lyons Laborator) Manager Kimberly A Evans TechnicalManager April 7,1994 1

9708190002 970813' PDR ADOCK 050004 0 j

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PY-CEl/0 PEA 0275L Page 2 of 9 StudyCbjective Daphnia magna bloassayp were conducted to present the detoxification potental of clays (Betz DT 1) for neutralizing the toxicity of Clam Trol CT 4. This evaluadon included ascertaining the neutralizing contribution of river water constituents by comprdng the detoxificadon potential of various cisy treaunents in both Detroit River wttar and i

labors:ory culture water. Laboratory culture water, with the exception of the clay addidons,is essentially devoid of any consdtuents 0.s., sils, suspended solids, hurr.ic acid) l

~~that would further and in detoxificadon In addition, this evaluation provided a comparison for the ' detoxification between, Clam ' Dol CT 4 and Clam Trol CT.1 using clays in Detroit River water Daphnia magne was selected as the test species for these bloassays due to its sensitivity to these molluscicidas.

-Methods

- Standard 48 hour5.555556e-4 days 0.0133 hours 7.936508e-5 weeks 1.8264e-5 months bloassays using Daphnfa magna were conducted in accordance with the -

EPA tasting procedure antitled

Methods icir Measuring Acuta Toxicity of Effluents and Receiving. Water to Preshwater and Marins Organisms", August,1993, "Ihree test series wers performed in this study, as.follows:

s Test Seria.it A range of clay treatments from 0 mg/L to 50 mg/L were applied to Clam Trol CT 4 test solutions prepared in Detroit A' lvar water All Clam Trol CT-4 test solutions were prepared at a concentration of 10 mg/L as product.

Test Series 2: A range of clay treatments from O mg/L to 50 mg/L were applied to Clam Trol CT-1 test solutions prepared in Detroit River water. All Clam *nol

~

,CT-1 test solutions were prepand at a concentration of 8 mg/L as product. + -

Test Sedes 5t A range of clay treatments fior'n 0 mg/L to 50 mg/L were applied to Clam Trol CT-4 test solutions prepared in laboratory culture water. All Clam.'

Trol CT 4 test solutions were prepared at a concentration of 10 mg/L as product.

Ihe Detroit River water was collected and shipped to Bett Aquatic Toxicology Laboratory overnight. The river water wu stored in a cold room until use. -

All test solutions for each test series were prepared in relplicates of two. Ten daphnids,

<24 hours old, were tr&nsiem:d to each test vessel and survival was monitond at 24 and

_48 hours.

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PY-CEI/0 EPA-02751.

Page 3 of 9 Results The mortality responses for each test series is pissented in the table of results. All Clam-l Trol CT-4 and CT-1 concentrations without any clay additions caused 100% mortality i

responses to the Daphnfa. The Clam-Tsol CT-4 test solutions (Test Series 1) in the dver wstor demonstrated a greater performance for detoalfdation than the CT 4 test solutions in the laboratory cultury water (Test Series 3). That is, a 10 mg/L CT-4 concentration in dvar water treated with 10 mg/L of clay (DT-1) ex' pressed only a 5% mortality, while the 10 mg/L CT-4 concentradon in culture water treated with 10 mg/L clay skpretted a 60%

mortality No mortality was expressed for the Cr.4 test solutions tasated with 20 mg/L of clay in either the dver water or culturs water. _ In summary, a treatment ratio of one part Clam-Trol Cf.4 to one or two parts clay (Betz DT 1) can adequately reduos or eliminate toxicity to Daphnis.

Test Series 2 presents the detoxification performance of clays for neutralizing Clam Trol CT 1. in comparison to Clam Trol CT-4, higher treatment ration of clay to CT-1 are required to achieve complete detoxification to the Daphnta. _8 mg/L CT-1 concentration treated with 30 mg/L of clay in river water (a treatment ratio of one part CT-1 to 3.75 parts as clays) eliminated toxicity.

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4

PY-CEl/0 EPA-02751.

Study ~Objectlye Page 4 of 9 l

Daphnia magna bloassay; were conducted to present the detoxification potential of clays (Batz DT 1) for neutralizing the toxicity of Clam Trol CT-4. This evaluation included ascertaining the neutralizing contribution of river water constituents by compadng the detoxincation potential of various clay tmatments in' both Detroit River water and i

i laboratory culture water. Laboratory culture water, with the exception of the clay

,,, additions, is essendally devold of any consdtuents (i.e., silts, suspended solids, humic acid) that would further aid in detoxification.

In addition, this evaluation provided a comparbon for the ' detoxification between. Clam Trol CT-4 and Clam-Trol CT 1 using' clays in Detroit River water. Daphn!a magna was selected as the test species for these bloassays due to its sensitivity to these rnolluscicides.

Methods l

l Standard 48 hour5.555556e-4 days 0.0133 hours 7.936508e-5 weeks 1.8264e-5 months bloassays using Daphnia magna were conducted in accordance with the EPA testing procedure entitled " Methods for Measuring Acute Toxicity of Etnuents and Receiving Water to Freshwater and Marine Organisms", August,1993. Three test series wem performed in this study, as follows:

Test Series.1: A range of clay treatments from 0 mg/L to 50 mg/L were applied to Clam Trol CT 4 test solutions prepared in Detroit River water. All Clam Trol CT-4 test solutions were prepared at a concentration of 10 mg/L as product.

Test Series 2: A nnge of clay treatments from O rug /L to 50 mg/L were applied to Clam Trol CT 1 test solutions prepared in Detroit River water. All Clam-Trol

.CT-1 test solutions were prepared at a concentration of 8 mg/L as product.

Test Setier 3: A range of clay tnatments frein 0 mg/L to 50 mg/L were applied to Clam-Trol CT 4 test solutions prepared in laboratory culttue water. All Clam '

Trol CT 4 ta.st solutions were prepared at a concentration of 10 mg/L as product.

The Detroit River water was collected and shipped to Betz Aquatic Toxicology Laboratory overnight. The river water was stored in a cold room until use.

All test solutions for each test series were papared in relplicates of two. Ten daphnids,

<24 hours old, were transferred to each test vessel and survival was monitored at 24 and 48 hours5.555556e-4 days 0.0133 hours 7.936508e-5 weeks 1.8264e-5 months .

4 4

breet Lakes Steel Study 1

e Delossicadon of ClanHrel CT-1 and CT '

[

caphnia aangna 48-Hr. %

l k

i Test Series 1

. Test Series 2 Test Sortes 3 CT-4 : Cieyi meer weser

% nsortamy CT-1 : Cimy/ Rher Water

% Atortamy CT-4 : CamyI Lab Water

% haortesy l

4 l

I

}

10-00 100 8:00 100 10-00 100 i

~

10.-10 5

8:10 100 10:10 80 10:20 0

8:20 75 10:20 0

i 10:30,

0 8:30 0

10:30 0

10:40.

0 8:40 0

10:40 0

.10:50 0

8:50 01 80:50 0

i i

0:50 '.

0 0:50 0

0:50 0

g t

)

i 10:0=10mg/L as CT-4 : No Cray 8:0=e.DmgT. as CT-1 : No Clay 10:1D=10mg4. as CT-4': 10 mg/t. Clay 8:10=6.0mpt as CT-1 : to rega. Clay i

1020=10mg/L as CT-4 : 20 mg/L Clay 820=8.0mgT as CT-1 : 20 ragt Clay

(

10:30=10mg4. as CT-4.: 30 mg1 Clay -

- 8:30=8.0mpt as CT-1 : 30,rugt Clay

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)

10:40=10mg/L as CT-4 : 40 mg/L Csay 8-40=8.OsngfL as CT-1 : 40 rag 4. Clay

gg 10:50=10mg/L as CT-4 r 50 mg4. Clay 8:50=8.0mgR. as CT-1 : 50 ragfL Clay

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0-50= 50 mg/L Clay only 0:50= 50 mgt Clay only R

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u Ckry formulation as Betz DT-1

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m Test Series 1 and 2 used Detroll River water Test Saries 3 used Laborakuy Culture water u

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PY CEl/0 EPA-0275L Page 6 of 9 CHRONIC EFFECTS OF DETOXIFIED CLAM. TROL CT.4 TO DAPHNEA MAGNA

Ihe eTg' cf of Betz DTG (a blend of bentonite clays) to midgate the chronic toxicity of Oam Trol CT-4 on survival, growth and esprodoodon of Daphnia magne was determined hem a 21 day lik. cycle toxicity. tast. D. mayes naamatan wars exposed to 25 mg/L Cism. Trol CT.4 combined with DTO clay levels of 0,25,37.5,50, and 75 mg/L Two controls consisting of untreated dilution. water and a 75 mg/L D113 test soludons were also assessed. Test achttions were answed every other day using moderately ~basd USEPA synthodo Deshwater as the dilution water. 1hs losicity test was inishead by placing one D. magna neonate in each test container, with a total of tan replicates per treatment. The chronic toxicity test methods employed fbuowed procedures; described by AS'rM (1939) Clam

Dol CT-4 and D10 levels were measured during each test solution renewal This ki day chronic toxicity test demonstrated the ability of aktz D1U clay to neutraHas the acute and chronic toxicitysassociated w{th Clam-Trol CT-4 to D. magna. !YIU clay fed at a level of 25 mg/L was an insufRcient amount of material to neutralian the toc;;,

of 25 mg/L Clam Trol CT.4. All daphnids died in the 25 mg/L D10 clay treatment because residual Clam Trol CT-4 levels measured in the test containers ' equaled 1.0 mg/L and were above the 1f50 or the molluscicide. The 48 hour5.555556e-4 days 0.0133 hours 7.936508e-5 weeks 1.8264e-5 months LC f

f 50 orD. magna is equal

{.

to 0.2 mg/L Clam TrolCT-4.

D1D clay levels equal to or greater than 37.5 mg/L offecdvely midgated the acuta and -

. chronic toxicity of 25 mg/L Clam.'nol CT-4. The rnolluscicide levels measured in these DTO treatments were below the 0.25 mg/L analytioni detection limit.

D. magna expressed a 50% monality response in the 37.5 mg/L DTU clay treatment during the first five days of the 21-day exposure, but was not alani5cantly different from the untmated control where 10% morulity occuned. The individuals which survived this tacatment.

grew larger and produced more neonate offspring than the untrasted control.

No impainnent to survival, growth and reproduction of D. magna was observed when exposed to 25 mg/L Clam Trol Cf 4 combined with DTG clay tretunents of 50 and 75 mg/L The LOEC (Lowest Observed Effect Concentration) for this chronic. toxicity test conesponded to the 25 mg/L DTG clay treatment combined with 25 mg/L Clam Trol CT-4. 'Ihe NOEC (No Observed Effect Conmotradon) was achieved when 37.5 mg/L DTG clay was combined with 25 mg/L Clam-Trol CT 4. 'Itere were no adverse effects observed to D. magna survival, reproduction or growth at any DTU clay treatments greater than 25 mg/L l

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W*G7 13846 8613 P.1C/32 PY-CE!/0 EPA-0275L Page 7 of 9 Clam.Tml DTG Cumulative Naonate Growth CT 4 Clay

(%)

Production Avg. Dry (mg/L)

(mg/L)

Mortality 4)

(+ S.E.M.) (b)

Wt. (mg) (*)

0 0

10 71.8 (8.8) 0.463 0

75 0

92.0 (4.6) 0.547 25 0

100

25 25 100 04EC) 25 37.5 50 (NOEC) 96.6 (9.1)(NOEC) 0.804 (NOBC) 25 50 0

112.8 (3.5) 0.851 25 75 0

103.1 (2.2) 0.874 l

a*. Signincantly different frore the entreated eentrol (= = 0.05, Fisher Exact Teet).

b. Neonate production, where depbmids survived, were not significantly different frera the entrentad control (as a 4.48, Wilanson rank seas test).

c Daphnid growth la all trestroents were not signifhetly different frees the entreated control

(= = 0.03, Bonferrval1 test).

'b 6

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.. -., e.w,u PV-CE l/0E PA-02751.

Page 8 of 9 CHRONIC EFFECIS OF DETOXIFIED CLAM TROL CT 4 ON FAT 11EAD MINNOW SURVIVAL AND GROM"TH

'Iha etnoiency of Ben DTU (a blend of boatonite days) to midgats the dronic toxicity of Cam.

'nol CT4 on unrvival, growth of Fathead minnow, Phnsphalarpromedar was datamalnad born a 7 day exposers. Patband mlanows less thna 24 home old were esposed to 25 ans/L Qam 'not.

CT4 conibland with 1710 day Icysts of 0, 25, 37.5, 50, and 75 mai. Two poemots naaetWag of untreated diludes water and a 75 mg& D10 day test saludoes were also ma====ad Test solutions was runswed evay day uslag moderately hard USEPA synthetic freshwater. as tha duution water. '!be loaldty test was iridated by pladas ten larval Ash lean four replicats test

=?* i with a total of 40 Sah suposed to each treatment. 'the ducale tosidty test namehade amployed fonowed procedmes descaibed by USEPA (1989). Qam Trol CT 4 and tyr0 clay i

levels were mensmed during andi test solution renewal.

This 7-day duconc toxicity test demonstrated the ability of Betz 17!0 day to neutralise the acute and chronic toaldty associated with Onm *not Cr4 to the fathead mlanow. All Eth died in the 25 m81 Osm.'Rol Cf 4 treatmaat within 24 hours2.777778e-4 days 0.00667 hours 3.968254e-5 weeks 9.132e-6 months in the shamaan of DTG clay. All DTU day treatment additions equal and greater than 2$ mg1 effe<xively mhigated the duordc toaldty of 25 mgA Osm-Trol Cf4 to the fathead mlanow. Thera was no antvival cir growth impairment expressed by the twval Osh when exposed for 7 days to 25 m:L Cam Trol CT4 combined with DTU clay treatments;n25 mg/L.

'!he LOBC (l.cwest Obearved Effect Coconnustion) for this toxidry test could not be determined -.

since the lowest DTO clay treatmeat combined with Qam.' Dol CT 4 did not exhibit 6ay advens effects to Ash anvival or growth. 'the NOEC (No Oboented E5ad t'==*ation) for this tent corresponded to the addition of 25 mg1 D10 day to neutrallee 25 mg1 Cam Trol CT-4 i

..n.

Oam Trol--

DTG Cumulative Growth Av3 CT4-Ony

(%)

Dry Wt. (mg)

(mgi)

- Mortality

+ (S.E.M.)

0 0

0.354 (0.014 0

75 10 0.525 (Oh43) 25 0

100*(LOEC) 25 25 12.5 (NOEC) 0.565 (0.040)(Norf) 25 37.5

'7.5 0.592(0.073) 25 50 10 0.596(0.033) 25 75 5

0.643(0.083)

sis.irie..uy dirr,e,.i trem ih.

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l(. o.as,si.d.M..y.o n..k Twi).

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- - - wo mses m i Hi&UNJ TO 4

2ig 290 geen II97eBG*07 13147

G13 P.21/32 PY-CE1/0 EPA-0275L Page 9 of 9 CHRONIC EFFECTS OF DETOXIFIED CLAM TROL CT 4

% SELRNASTRUM CANtlCORNU111M The of8ciency of Bett DE clay to mitigate the chronic toxleity of Clam Trol Cr-4 on the grow & of a green alsa,'s+ mw espricomunas, was determined from a 5 hour5.787037e-5 days 0.00139 hours 8.267196e-6 weeks 1.9025e-6 months stanc impairment test. De algas were exposed to 25 mg/L Qam.* Dol CT-4 combined with DTO clay levels of 0,15,37.5,50, and 75 mg/L, All Qam Trol Cr4 and DTO clay test solutions wem prepared in USEPA algal modia as ths test duusat Two controls consisting of untreated algal media and a 75 mg/L D'IU clay were also asasseed in the l

toxicity test.

Test chambers were sterus 1251n1 Brlemayer Danka containtag $0 ml of test saludon.

l Plasks were inoculated with 1 ad of acck algal cultus which had been adjusted to a ceu coneantration of approvimately 5.0 X 105 cons /ml. Each treatment consisted of thma replicate Banks containing 1 X 10' A osuartal. Test Rads were incubated on a rotary shaksr at 100 rpm under continuous Ulaminados of 400 A 40 foot candles. De test soludons were mainmined at 25 A PC. De chrorde algal towth test methods employed S

followed those outlined in USEPA (1939). At the end of the 5 hour5.787037e-5 days 0.00139 hours 8.267196e-6 weeks 1.9025e-6 months test, algal cell densides in each test flask were measured as a relative absorbance uslag a Turner Pluorornstar.

ihls 96-hour chronic algal growth impahment test demonstrated the abiuty of Betr. DTO clay to neutralize the toxicky associated with Qam Trol CT-4 to Selenasoum capricornunun. Based on comparison with reladve absorbance of untreated controla, -.

growth of Selenanrum eqpricomems was signl5 candy impaired in the unannant with 25 m3/L Qam Trol CT-4 alone, and with the addidon of 25 mg/L 17FO clay, he LOEC (Lowest Observed Effect Concentration) for this test cormsponded to the 25 mg/L D'lX) clay treatment combined with 25 mg/L Clam ' Dol CT-4. Convemelf~ 25 mg/L Qam Trol CT 4 was detoxified with DM clay added at levels greater than 37.5 mg/L.

%erefore, the NOEC (No Observed Effect Concentration) for this test corresponded to the addidon of 37.5 mg/L DTO clay to neutrallac 15 mg/L Clam-Tml CT-4. Exposure to 75 mg/L DTG clay alone had no significant impact on algal growth.

Average Cam Trol DTG Relative CT.4 Qay Absorbance (mg/L)

(mg/L) at 96 Hrs.

0 0

20.17 0

75 2183 25 0

0.01

25 25 17.3 * (LOEC) 25 37.5 19.17 (NOEC) 25 50 18.83 25 75 20.13

447 dy dirrerent from the wattgated control (n = 0.05, Dunnett Test)

ML20210J873

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