Document 93LrpNwQOr6m7gq096RGdnKxD

K7l.>9-23 Htuluiiual BIO-TEST Jta/xMto'ueS; Unc. 1810 FRONTAGE ROAD NORTHBROOK, ILLINOIS 60062 AftCA cooc m TllMHONI 173 -1010 REPORT TO MONSANTO COMPANY FOUR-DAY FISH TOXICITY STUDIES ON SEVEN MATERIALS IBT NO. A7204 sUc MUNi 089044 2tfdu&t/Ual 010 * T E S T J!abo>iat(>>ue&, J)nc, 1810 frontage road NORTHBROOK. ILLINOIS 60062 AM* COOC III mi.HONl Itt-IOIA June 13, 1969 Mr. Elmer P. Wheeler, Manager Environmental Health Monsanto Company 800 N. Lindbergh Boulevard St. Louis, Missouri 63166 Dear Mr. Wheelers Res XBT No. A7204 - Four-Day Fish Toxicity Studies on Seven Materials We are submitting herewith our laboratory report dated June 13, I969 prepared in connection with the above study. Very truly yours. JCC/kjl J. C. Calandra President MONS 089045 SluLtUnlal D I O T E S T JaiavUiyUM. }no. REPORT TO MONSANTO COMPANY FOUR-DAY FISH TOXICITY STUDIES ON SEVEN MATERIALS IBT NO. A7204 I. Introduction At the request of Monsanto Company, four-day fish toxicity studies in Rainbow Trout (Salmo gairdnerii) and Bluegills (Lepomis macrochirus) were conducted on the following seven materials: 1) Aroclor 1242, Lot 0AK-255 2) Aroclor 1254, Lot #AK-3B - Cr 'S\t * S' 3) Aroclor 1260, Lot #AK-3 C.P 9* 1 4) Aroclor 5460, Lot HAK-390 c? U3?i 5) Toxapheiie ' ' 6) Technical DDT (2-1/2#), B0903604 7) Halo,Wax\1099, 52 percent Cl MQNS 089046 MtilUlal D I O - T E S T JaiaialtMei, Sue. 2 II. Summary The results of the four-day fish toxicity studies are summarized below. Due to the paucity of one of the seven test materials (Halowax 1099). complete data are not available and the figures in parentheses represent only range-finding data. Test Material Four-Day LCjq (ppm) Rainbow Trout Bluegill Aroclor 1242 > 100.0 0.66 Aroclor 1254 > 100.0 0.66 Aroclor 1260 - >100.0 > 100.0 Aroclor 5460 > 100.0 > 100.0 Toxaphene 0.0080 . 0.0056 Technical DDT 0.0052 ' 0.0078 Halowax 1099 (10-100) (> 100.0) Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. Report prepared by: AAvx^vj Carmen Mastri, B.S. Section Head Acute Toxicity Department ' Report approved by: M. L. Keplinger, Ph.D. Assistant Laboratory Director June 13, 1969 Otis E. Fancher, Ph.D. Director MOWS 06904 7 JnJitttUal BIO-TEST Jaiowivtici. }ne. 3 III. Procedure The procedure was generally the same for both blucgills and rain bow trout with each test material. Healthy, young fish with an average length of 35 to 75 millimeters were used as test animals. All fish were kept under observation for a period of not less than ten days prior to experimental use. During this period, the fish were observed for general health and suitability as test animals. The fish were held in large aerated stock tanks and were fed frozen brine shrimp until three days immediately prior to testing. The rainbow trout were maintained at a temperature of 13C and the blucgills at a temperature of 24C. All stock tanks and bioassay vessels contained reconstituted, deionized water. The following compounds were added in the amounts stated per liter of deionized water: 30 mg calcium sulfate , 30 mg magnesium sulfate ' 48 mg sodium bicarbonate 2 mg potassium chloride Preliminary screening was conducted in order to find the general level of toxicity before beginning the actual bioassay. Once this level had been determined one to five concentrations were selected for the actual bioassay. Bioassay vessels lined with disposable polyethylene bags were filled with ten liters of well aerated reconstituted water. To evaluate the selected concentrations, ten trout were placed in each bioassay vessel. In the case of the blucgills only five fish were placed in each bioassay vessel in order to insure adequate dissolved oxygen concentration (D. O.) MONO 089040 fniiiitUal B I O - T E S T lahyiaiouM, Due, 4 In the warmer water; therefore two groups of five bluegilla were tested at each selected concentration. An untreated control group consisting of 10 fish was also employed. After a 24-hour acclimatization period, calculated amounts of test material necessary to produce the desired one to five concentration levels were dispensed into the respective vessels. The test solutions were prepared within one hour of the bioassay. , Following the introduction of the test material the fish were observed for 96 hours during which time all mortalities and/or untoward behavioral reactions were recorded. The concentration of dissolved oxygen was de termined for all test solutions in which mortalities occurred, to insure that adequate oxygen was available to the test specimens*. Concentrations Of dissolved oxygen of 4 mg/1 (4 ppm) for warm water fish (bluegill) and 5 mg/l (5 ppm) for cold water fish (trout) were considered to be minimal for valid testing. The pH of the test solution was also determined when mortalities were recorded. At the end of the observation period all data were collected and ar rangements were made to calculate the 96-hour LCSo employing the technique of Litchfield and Wilcoxon**. * Standard Methods for the Examination of Water and Wastewaters. Twelfth Edition, American Public Health Association, Inc., 1790 Broadway, New York, N. Y.: Part HI, pp. 406-410. ** J. T. Litchfield, Jr. and F. Wilcoxon. "A Simplified Method of Evaluating Dose-Effect Experiments,1* J. Pharm. M Exp, Thcr. , Vol. 96, No. 2. pp. 99-113. MOMS 089049 9ndtUlal BIO-TEST Jahowktloi, Jruk 5 IV. Results No deaths or untoward behavioral reactions were noted among any of the animals in the untreated control groups. A. Rainbow Trout 1. Aroclor 1242, Aroclor 1254, Aroclor 1250, AtqcIqt 5460. Toxaphene, Technical DDT, Halowax 1099* ~" a. Mortality, DO, and pH Data --. : . The mortality, dissolved oxygen content, and pH data are presented in Tables I through IV. ... ... , * Range finding mortality only. MOWS 0B9050 ISOfcflO SNOW wy wrmep'r i s a 1 1qo "ST Four-Day Fish Toxicity Study - Rainbow Trout Mortality, DO, and pH Data , Number Dead Total Per Dissolved Oxygen (ppm) and pH V Concentration* Hours: Number Number Cent 1-6 Hr. 24 Hr. 48 Hr. 72 Hr, 96 Hr. Test Material (ppm) _ 1-6 24 48 72 96 Dead Tested Dead DO pH DO pH DO pH DO pH DO pH Aroclor 1242 100.0 0000 0 10 Aroclor 1254 100.0 0000 0 10 Aroclor 1260 Arbclor S4(>(f 100.0 0000 So /o <o /o 0 10 --io---- Pour-Day LC50 Aroclor 1242 > 100. 0 ppm Aroclor 1254 > 100.0 ppm Aroclor 1260 > 100, 0 ppm ^Aroclor 546Q > .200, 0 ppmy*} ' * i ; i * The test materials were dispensed Into the bioassay vessels in the form of a 10.0 percent (w/v) solution in acetone. Concentrations are expressed in terms of actual test material as received. O' TABLEn TEST MATERIAL: Toxaphene Four-Day Fish Toxicity Study - Rainbow Trout Mortality, DO, and pH Data Number Dead Total neentration* Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested Percent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 1 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 0.0056 00000 0 10 0 . 0.010 09000 9 10 90 - - 8.2 7.4 - -. .- _ 0.018 0 9 1 ' - - 10 10 100 - - 8.2 7.6 8.0 7.4 - -- _ 0.032 0 4 6 - - 10 10 100 - - 7.8 6.9 8.1 7.0 - -- 0. 056 0 10 - - - 10 1 ; 100 . ,, 8.1 7.2 . _ ur-Day LC50 = 0.008 ppm . The test material was dispensed into the bioassay vessels in the form of a 0.01 percent (w/v) solution in acetone. Concentrations are expressed in terms of actual Toxaphene as received. * c sr c OB <> CVJi fsj VoflV 'H o-pr 1 S 3 1 - O JO V * `P >,F ,S -si w \ TABLE HI TEST MATERIAL: Technical DDT Four-Day Fish Toxicity Study - Rainbow Trout' Mortality, DO, and pH Data Number Dead Total :cr.trat ion* Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested 0. 001 000 00 0 10 Per Cent 1-6 Hr. Dead DO pH 0 Dissolved Oxygen (ppm) and pH ' 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO PH DO pH DO pH DO pH 0.0018 0.0032 0 0 0 0' 0 0 0 0' 0 0 0 0 10 10 0 0 _ 0.0056 0.01 0 0 4' 1 2 063 1 - 7 10 10 10 ' 70 100 .- -- - - 8.0 7.3 7.8 7.2 8.4 7.2 8.2 7.3 8.0 7.2 7.9 7.2 - - -Day LCgg = 0.0052 ppm le test material was dispensed into the bioassay vessels in the form of a 0. 001 percent (w/v) solution in acetone, jncentrations are expressed in terms of actual Technical DDT as received. 3 (A o a Otji I* is 31*010 oo < V Jl S 3 i * 0 IB r 'W V 't TABIX IV TEST MATERIAL: Halowax 1099 Range Finding Fish Toxicity Study - Rainbow Trout- Mortality, DO, and pH Data Number Dead Total * it rat ion" Hours: Number Number pm) 1-6 24 48 72 96 Dead Tested Per Cent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH ' 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 000 2 2 2 100 - - 10.0 00 00 0 0 2 0 -- Day LC50 = 10-100 ppm : test material was dispensed into the bioassay vessels in the form of a 10,0 percent (w/v) solution in acetone, icentrations are expressed in terms of actual Halowax 1099 as received, ' : 0 1 0 r~ z c ! 2 ca xoO' <0 Co jpttkuU^Ual DIO-TEST Pnc. Jt 10 b. Reactions The reactions noted following introduction of the Aroclor 1242, Aroclor 1254, Aroclor 1260, Aroclor 5460, Toxaphcne, end Technical DDT are presented in Tables V through X respectively. MGNS G89055 P ftJ itiU ia l B I O - T E ST Jahviaio'UeS, Shc. TABLE V TEST MATERIAL* Aroclor - 1242 Four-Day Fish Toxicity Study - Rainbow Trout . Summary of Reactions Concentration (PPTM> Reaction 100.0 Hypoactivity Inverted Swimming Time of Onset Following Dose Administration (hours) Duration of Reaction (hours) Time o Death Following Dose Administration (hours) 24 No Recovery Noted 72 No Recovery Noted - j , 1 l c 2/- C a *0 O \j> a U 0S tu lu iltfa l B I - T E ST ~ah>UiIosUoS, 5hc. TABLE VI TEST MATERIAL: Aroclor - 1254 Four-Day Fish Toxicity Study - Rainbow Trout Summary of Reactions Concentration (ppm) Reaction 100.0 No Reactions Noted Time of Onset Following Dose Administration (hours) - Duration of Reaction (hours) - Time of Death Following Dose Administration (hours) - xt o2w c00 oLT M w ytubtifotol B I O - T E ST MUNS 08905a TABLE TO TEST MATERIAL! Aroclor - 1260 Four-Day Fish Toxicity Study - Rainbow Trout Summary of Reactions Concentration (ppm) 100.0 Reaction Hypoactivity Time of Onset Following Cose Administration (hours) Duration of Reaction (hours) Time of Death Following Dose Admini s tration (hours) 72 No Recovery Noted - / l. TABLE VIH ' ' 1 TEST MATERIAL: Aroclor - 5460 J Four-Day Fish Toxicity Study - Rainbow Trout _ Summary of Reaction* ---- -- Concentration (PP*n) 100.0 Reaction No Reactions Noted Time of Onset Following Dose Administration (hours) Duration of Reaction (hours) Time of Death Following Dose Administration . Jhours) *" |. V ^ g, CJ -- * m m VI ^ . | f V TABLE DC TEST MATERIAL: Toxaphene Four-Day Fish Toxicity Study - Rainbow Trout Summary of Reactions Concentration (com) Reaction 0. 0056 No Reactions Noted Time o1 Onset Following Dose Administration (hours) '- 0.010 Hypoactivity 24 0.018 Hyperactivity , 24 0. 032 0. 056 Hyperactivity Hyperactivity . 24 1-6 Duration of Reaction (hours) Time of Death Following Dose Administration (hours) -24 24 Until Death Until Death Until Death 24-48 24-48 24 / c z Ui Ca <> o O B IO -TE S T in W TABLE X TEST MATERIAL: Technical DDT Four-Day Fish Toxicity Study - Rainbow Trout Summary of Reactions Concentration (ppm) Reaction 0. 001 No Reactions Noted 0. 0018 No Reactions Noted 0. 0032 No Reactions Noted 0. 0056 Hyperactivity 0. 01 Hyperactivity Prostration Tune of Onset Following Dose Administration (hours) '- - ; i-6 . 1-6 Duration of Reaction (hours) - Time of Death Following Dose Administration (hours) _ -- -- No Recovery Noted Until Death 48-72-96 24-48-72 2 Oz (/> c (X -c o O' w y to n o /v x y e r X s 3 i - 0 1 8 p 'r'?f"f u( u SiutiulMal BIO-TEST Jal&ioi&ud, }*c. 17 B. Blucgllls 1. Aroclor 1242, Aroclor 1254, Aroclor 1260, Aroclor 5460 Toxaphcne, Technical DDT and Halowax 1099* a. Mortality, DO, and pH Data The mortality, dissolved oxygen content, and pH data aro presented in Table XI through XVII respectively. I * Range finding mortality only. MUNS 089062 TABLE XI TEST MATERIAL: Aroclor 1242 Four-Day Fish Toxicity Study - Bluegill Mortality, DO, and pH Data Number Dead Total ' :cntration* Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested Per Cent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 0.10 0 00 00 0 10 0 0.18 0 00 00 0 10 0 0.32 0 0000 0 10 0 0.56 1.0 0 0 0 0. 0 0 10 - 0 10 10 0 10 100 7.6 7.2 - r-Day LCsq = 0. 66 ppm ` be test material was dispensed into the bioassay vessels in the form of a 1.0 percent (w/v) solution in acetone, oncentrations are expressed in terms of actual Aroclor 1242 as received. x oz c a o O B I O - T E ST JakyuilytUS, (% TABLE XU - TEST MATERIAL: Aroclor 1254 Four-Day Fish Toxicity Study - Bluegiil Mortality, DO, and pH Data Number Dead Total :cntration* Hours: Number Number (ppm) 1 6 24 48 72 96 Dead Tested Per Cent . 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 0.10 0. IS 0.32 0.56 1.00 0 0000 60000 000 0 0 0 0 0 0- 0 0. 0 6 3 1 0 0 0 0 10 10 0 - 10 0 - - * 10 0 - - 10 0 - - 10 100 -- - - .7.8 7.2 6.5 7.2 6.2 7.1 r-Day LC5Q 0. 66 ppm * he test material was dispensed into the bioassay vessels in the form of a 1,0 percent (w/v) solution in acetone, oncentrations are expressed in terms of actual Aroclor 1254 as received. x c z <X O o a* V 1 S3 1 - 0 MI P W f U b 0}n d u ib la l BI TEST Jai& 'uiU 'ilcA , Una. TABLE XIH TEST MATERIAL: Aroclor 1260 Four-Day Fish Toxicity Study - Bluegill Mortality, DO, and pH Date Number Dead Total ncentration Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested Per Cent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO oH 100.0 00000 0 ` 10 0 -- ir-Day LCso> 100.0 ppm ' Hie test material was dispensed into the bioassay vessels in the form of a 10. 0 percent (w/v) solution in acetone. Concentrations are expressed In terms of actual Aroclor 1260 as received* - * cz / <s. c CD sC o o- VI otv fn iu iU k d B I O - T E ST X ahyw iotiU , $nc. TABLE XIV TEST MATERIAL: Aroclor 5460 Four-Day Fish Toxicity Study - Bluegill Mortality, DO, and pH Data Number Dead Total mccntration* Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested Percent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 100.0 00000 0 10 0 -- wr-Day LC$q> 100,0 ppm ` , The teat material was dispensed into the bioassay vessels in the form of a 10,0 percent (w/v) solution in acetone. Concentrations are expressed in terms of actual Aroclor 5460 as received. 3/ Cr is. ca c Oo' csj tu }nJuil*Jal B I O T EST Jtd-i'ux/o'Uei. Hue. MGNS 0 8 9 0 6 7 . TABLE XV i TEST MATERIAL: Toxaphene 1 Four-Day Fish Toxicity Study - BluegUl Mortality, DO, and pH Data Number Dead Total iccntration* Hours: Number Number (ppm) 1-6 24 48 72 96 Dead Tested Per Cent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 0.0010 00000 0 10 0 0.0018 00000 0 10 0 0.0032 00000 0 10 0 0.0056 0.01 0 0 5 0. 0 0070 1 5 8 10 10 50 80 - . - - - 7.3 7.2 - - 7.3 7.2 - -- - - 6.1 7.2 r-Day LC5Q = 0,0056 .- ' percent Confidence Limits of LC50 = 6.0036 - 0.0087 ppm The test material was dispensed into the bioassay vessels in the form of a 0,01 percent (w/v) solution in acetone. Concentrations are expressed in terms of actual Toxaphene as received. Mtv f TABLE XVI. TEST MATERIAL: Technical DDT Four-Day Fish Toxicity Study - Bluegill Mortality, DO, and pH Data Number Dead Total ;cr.trat ion1* Hours: Number Number (ppm) I -6 24 48 72 96 Dead Tested Percent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO dH 0.001 0.0018 0 000 0 .0 0 0 0 0 0 0 10 10 0................................................................................... 0 . ......................................................................... ......... 0.0032 0 000 0 0 10 0 . ...................................................................................... 0.0056 0 0 0 0. 0 0 10 o'..................................................................... 0.01 0 10 - - - - 10 100 - - 7.6 7.1 - -- -- - c* 1 C3 O m <n t -Day LCgQ * 0,0078 ppm *t . ie test material was dispensed into the bioassay vessels in the form of a 0.001 percent (w/v) solution in acetone, jncentrations are expressed in terms of actual Technical DDT as received. ox z Co c a *6 Oa a utv }ndniUlal BIO -TEST Jc, TABUS xvn TEST MATERIAL: Halowax' 1099 Range Finding Fish Toxicity Study - BluegUI Mortality, DO, and pH Data Number Dead Total ' cntratlon* Hours: Number Number ppm) 1-6 24 48 72 96 Dead Tested Per Cent 1-6 Hr. Dead DO pH Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hr. 72 Hr. 96 Hr. DO pH DO pH DO pH DO pH 100.0 00000 0 2 0 10.0 00000 0 2 0 -Day LC50 > 100, 0 ppm e test material was dispensed into the bioassay vessels in the form of a 10. 0 percent (w/v) solution in acetone, ncentrations are expressed in terms of actual Halowax 1099 as received. o1 c a>c oa 9ttditiblal B I O T E S T Jalm.'iattyUcl, Sue, 25 b. Reaction The reactions noted following Introduction of Aroclor 1242, Aroclor 1254, Aroclor 1260, Aroclor 5460, Toxaphene, and Technical DDT are presented in Tables XVIII through XXIII respectively. MUNS 0890 70 0 6 B0 SNOWU x S 31 0 I a vrvP 'T ut TABLE XVIH TEST MATERIAL: Aroclor - 1242 Four-Day Fish Toxicity Study - Bluegill Summary of Reactions Concentration (pom) Reaction Time of Onset Following Dose Administration (hours) 0.1 No Reactions Noted ' - 0.18 0.32 No Reactions Noted Hyperactivity , -` 1-6 0. 56 Hyperactivity 1-6 1.0 Hyperactivity ' 1-6 Duration of Reaction (hours) - Time of Death Following Dose Administration (hours) . - -' - . 24 24 Until Death 24 / in J ttlU ia l B I O - T E ST Jalota&yUei, Snc. TABLE XIX 1 TEST MATERIAL: Arodor - 1254 Four-Day Fiah Toxicity Study - BluegiU Summary of Reactions Concentration (ppm) Reaction Time oX Onset Following Dose Administration (hours) Duration of Reaction (hours) Time of Death Following Dose Administration (hours) 0. 1 No Reactions Noted ' -' - - 0.18 No Reactions Noted - -- 0.32 0.56 No Reactions Noted , Hyperactivity Hypoactivity - .1-6 24 . -. 24 48 - - 1.0 Hypoactivity 24 Until Death 48-72-96 / ? t 0 6 9 0 SNOW IV fc TABLE XX . TEST MATERIAL: Aroclor - 1260. I . Four-Day Fish Toxicity Study - Bluegill Summary of Reactiona Concentration ' (Bum) Reaction 100.0 Hypoactivity i* Time of Onset Following Dose Administration (hours) Duration of Reaction. (hours) Time of Death Following Dose Administration ,' 48 No Recovery Noted ! us O m, z cz / c a Ovl U (V 00 *S `w yo 7 v i S 3 I O I 0 P'.'V' T ' f TABLE XXI TEST MATERIAL: Aroclor - 5460 Four-Day Fish Toxicity Study - Bluegill : Summary of Reactions Concentration _____<PPm) Reaction 100.0 Hypoactivity . Time of Onset Following Dose Administration (hours) Duration of Reaction (hours) Time of Death Following Dose Administration (hours) 72 No Recovery Noted - 2 C (zA / c a < O rv o >,uU U M al B l O - T E ST Jc MUNS 0 8 9 0 7 5 TABLE XXH TEST MATERIAL: Tcxaphene Four-ray Fish Toxicity Study - Bluegill Summary of Reaction* Concentration (ppm) Reaction 0.001 No Reactions Noted 0.0018 No Reactions Noted 0.0032 No Reactions Noted 0. 0056 Hypoactivity 0.01 Hypoactivity Prostration Loss of equilibrium Time of Onset Following Dose Administration (hours) '- Duration of Reaction (hours) - Time of Death Following Dose Administration (hours) ' - '- - -- ' 48 24 48 24 ` 24 48 Until Death . 48-96 oW Sr TABLE XXUI TEST MATERIAL; Technical DDT ' Four-Day Fish Toxicity Study - Bluegill Summary of Reaction# Concentration _____(PP") Reaction 0.001 0. 0018 0.0032 0. 0056 0.01 No Reactions Noted No Reactions Noted No Reactions Noted No Reactions Noted Hyperactivity Time of Onset Following Dose Administration (hours) Duration of Reaction (hours) Time of Death Following Dose Administration (hours) '' - _ - .. .- - - .- - ' .' - -- 1-6 Until Death 24 / c is, o Cl >C o * V I S3i - O I 0