EPA-600/3-76-061b July 1976 Ecological Research Series STANDARD TEST FISH DEVELOPMENT Parti Environmental Research Laboratory Office of Research and Development U.S. Environmental Protection Agency Duluth, Minnesota 55804 ------- RESEARCH REPORTING SERIES Research reports of the Office of Research and Development, U.S. Environmental Protection Agency, have been grouped into five series. These five broad categories were established to facilitate further development and application of environmental technology. Elimination of traditional grouping was consciously planned to foster technology transfer and a maximum interface in related fields. The five series are: 1. Environmental Health Effects Research 2. Environmental Protection Technology 3. Ecological Research 4. Environmental Monitoring 5. Socioeconomic Environmental Studies This report has been assigned to the ECOLOGICAL RESEARCH series. This series describes research on the effects of pollution on humans, plant and animal species, and materials. Problems are assessed for their long- and short-term influences. Investigations include formation, transport, and pathway studies to determine the fate of pollutants and their effects. This work provides the technical basis for setting standards to minimize undesirable changes in living organisms in the aquatic, terrestrial, and atmospheric environments. This document is available to the public through the National Technical Informa- tion Service. Springfield, Virginia 22161. ------- EPA-600/3-76-06lb July 1976 STANDARD TEST FISH DEVELOPMENT PART II Chronic Toxicity of Guthion to the Fathead Minnow (Pimephales promelas Refinesque) by Ira R. Adelman and Lloyd L. Smith, Jr. Department of Entomology, Fisheries, and Wildlife University of Minnesota St. Paul, Minnesota 55108 Grant No. R800940 Project Officer Robert W. Andrew Environmental Research Laboratory Duluth, Minnesota 55804 U.S. ENVIRONMENTAL PROTECTION AGENCY OFFICE OF RESEARCH AND DEVELOPMENT ENVIRONMENTAL RESEARCH LABORATORY DULUTH, MINNESOTA 55804 ------- DISCLAIMER This report has been reviewed by the Environmental Research Laboratory - Duluth, U.S. Environmental Protection Agency, and approved for publication. Approval does not signify that the contents necessarily reflect the views and policies of the U.S. Environmental Protection Agency, rior does mention of trade names or commercial products constitute endorsement or recommen- dation for use. ii ------- ABSTRACT (£> Three chronic bioassays of Guthioir^were conducted with fathead minnows. All tests were begun with eggs and the longest lasted 20 days after termination of spawning, a total of 250 days. Parameters measured were survival, growth, fecundity, and growth and survival of second genera- tion fry and eggs. The most sensitive criteria for effect of the toxicant was fecundity. The maximum acceptable toxicant concentration (£> was between 0.33 and 0.51 pg/liter Guthion, and the application factor was between .0017 and .0027 based on a soft water acute and a hard water chronic bioassay. iii ------- CONTENTS Page Abstract iii List of Tables vi Acknowledgements V11 Sections I Conclusions 1 II Recommendations 2 III Introduction 3 IV Materials and Methods 4 Number of Experiments 4 Apparatus 4 Fish 5 Chemical Analysis 9 V Results 11 ( Egg and Fry Survival 11 Growth 13 Reproduction 13 Second Generation 16 VI Discussion 17 VII References 18 VIII Publications 20 IX Glossary 21 ------- TABLES No. Page 1 Analysis of Well Water 6 2 Means and Standard Deviations (in Parentheses) of Conditions in All Test Chambers 7 3 Percentage Survival of Fathead Minnow Eggs and Young Exposed to Various Concentrations of Guthion^ 12 4 Mean Weight of Fathead Minnows at Various Time Intervals After Spawning (Number of Fish Weighed in Parentheses) 14 5 Fecundity of Fathead Minnows Exposed to Various Concentrations of Guthion^ 15 6 Effect of Guthiori^on Survival and Growth of Second Generation Eggs and Fry 16 vi ------- ACKNOWLEDGEMENTS The authors wish to thank Richard W. Frenzel and particularly Gary D. Siesennop for assistance in conducting the experiments. vii ------- SECTION I CONCLUSIONS The results described in the following report permit certain conclusions concerning the toxicity of Guthiorr to fathead minnows. 1. The maximum acceptable toxicant concentration for fathead minnows is between 0.33 and 0.51 yg/liter Guthion. 2. The application factor comparing an acute bioassay in soft water and a chronic bioassay in hard water is between .00017 and .00027. ------- SECTION II RECOMMENDATIONS (iv) Guthion^ concentrations in the environment should not exceed 0.33 yg/liter. ------- SECTION III INTRODUCTION GuthiorP (azinphosmethyl) is an important and widely used insecticide on deciduous fruits and cotton as well as many other fruits and vege- tables. Although acute toxicity data have been reported by many workers for a variety of fish species (Katz, 1961; Pickering, Henderson, and Lemke, 1962; Meyer, 1965; and Macek and McAllister, 1970), no informa- tion has been reported on the effect of exposures over a complete life cycle. Since data on chronic toxicity are important for establishment of water quality standards, fathead minnows were exposed to various ® concentrations of Guthion over a complete life cycle, from egg through fry in the second generation, in order to determine a maximum acceptable toxicant concentration (MATC) (Mount and Stephan, 1967). ------- SECTION IV MATERIALS AND METHODS NUMBER OF EXPERIMENTS Three experiments were conducted, subsequently referred to as tests I, II, and III. Tests I and II were preliminary and were terminated after 26 and 68 days, respectively, when it became apparent that the lowest concentration was causing significantly greater mortality than the control. Furthermore chemical analysis of small Guthion concentrations had not been perfected, and residues on glassware and water sampling equipment made analyzed concentrations inaccurate. This problem was particularly true for test II where desired concentrations were low enough to be influenced. Therefore, information from the first two tests is only roughly quantitative but does indicate an area of possible effect. For test III all glassware was soaked in a strong sodium hy- droxide solution (pH = 12"^ for at least two days prior to use and no residues were found in blank analyses. APPARATUS A proportional diluter (Mount and Brungs, 1967) that dispensed seven toxicant concentrations with a 60% dilution factor and one control was used for administration of the toxicant in a flow-through system. Technical grade Guthioir^ (93%) was added directly to a single mixing chamber by the injector described by DeFoe (1975). An air dispersion stone and baffles thoroughly mixed the Guthion^with incoming water before it was siphoned to the toxicant cells in the diluter. The test chambers for first generation young and adult fish were constructed of ------- glass and G.E. Silicone Seal, measured 50 x 25 x 20 cm, and contained 20 liters. With a flow rate of 250 ml/min, 90% replacement of water occurred in approximately 3-1/4 hr. Temperature was maintained at a constant 25 C by a hot water exchange system in the head tank where aeration also occurred. All test chambers in test III contained an air dispersion stone in order to maintain oxygen concentrations. A dense bacterial growth occurred in the two highest concentrations apparently using the Guthion^ as a nutrient and the aeration was necessary to replace oxygen consumed by the bacterial respiration. Chambers were siphoned daily except when fry were present to keep this bacterial growth to a minimum. A complete analysis of hard water from a deep well is reported in Table 1. Those water quality characteristics which varied in individual test chambers are listed in Table 2. Fry in the second generation were reared in glass chambers measuring 20 x 20 x 21 cm and containing 6.4 liters. For each concentration water flowed from the outlet of the adult test chamber into the first fry chamber, then from the outlet of that to a second fry chamber so that two groups of offspring from the same adults could be reared simul- taneously. Eggs were hatched in a small jar with a nylon screen bottom. One jar in each adult test chamber was oscillated so that water flowed past the eggs without raising them from the screen. All adult chambers were exposed to the direct illumination of a 40-watt cool white fluorescent tube, suspended 50 cm above the water surface. Second generation fry chambers received only indirect illumination from laboratory light. Tests were begun with a photoperiod equivalent to January 1 at Evansville, Indiana and adjustments were made every 15 or 16 days. FISH Fathead minnows originally obtained from the Environmental Research Laboratory at Duluth, Minnesota were reared in a laboratory culture system. All tests were started with eggs from five spawning pairs which ------- Table 1. ANALYSIS OF WELL WATER (milligrams/liter) Item Concentration Total hardness as CaCO, Calcium as CaCO_ Iron Chloride Sulfate Sulfide Fluoride Total phosphates Sodium Potassium Copper Manganese Zinc Cobalt* nickel Cadmium, mercury Ammonia, nitrogen Organic nitrogen 220 140 0.02 <1 <5 0.0 0.22 0.03 6 2 0.0004 0.0287 0.0044 <0.0005 <0.0001 0.20 0.20 taken from well head before aeration and heating; pH 7.5. ------- Table 2. MEANS AND STANDARD DEVIATIONS (IN PARENTHESES) OF CONDITIONS IN ALL TEST CHAMBERS Item Test chamber 12345678 Guthiori^Xug/l) 0 7.6(1.3) Temp. (C) 24.8(.6) 25.0(.6) pH (reading) 7.73(.13) 7.53(.11) DO (mg/1) 6.60(.66) 5.95(.24) Guthion®(yg/l) 0 1.8(.9) Temp. (C) 24.6(1.7) 24.6(1.8) pH (reading) 7.75(.09) 7.58(.11) DO (mg/1) 6.28(.77) 5.54(.45) Guthion®(yg/l) .04(.OS)-'' .10(.04) Temp. (C) pH (reading) DO (mg/1) 24.1(1.1) 24.4(1.2) 6.96(.34) 6.52(.57) Test I 11.7(1.9) 20.5(1.9) 34.5(3.2) 24.9(.6) 25.0(.6) 25.0(.6) 7.52(.10) 7.49(.09) 7.51(.ll) 5.55(.56) 5.03(.05) 5.20(.74) Test II 1.9(1.0) 2.8(1.1) 3.4(1.2) 24.6(1.9) 24.4(1.9) 24.6(1.9) 7.57(.10) 7.60(.09) 7.60(.09) 5.55(.45) 5.58(.31) 5.36(.38) Test III Adult chambers .16(.05) .24(.06) .33(.08) 24.4(1.2) 24.4(1.2) 24.6(1.4) 8.07(.ll) 8.06(.ll) 8.05(.ll) 6.49(.34) 6.27(.64) 5.79(.59) 61.4(2.4) 96.7(6.6) 165.9(13.1) 24.9(.6) 25.1(.6) 25.1(.6) 7.52(.16) 7.50(.09) 7.56(.17) 4.48(1.28) 4.46(.61) 5.13(.51) 4.8(.8) 6.5(.8) 15.0(2.2) 25.4(.6) 25.3(.6) 25.4(.6) 7.58(.08) 7.60(.09) 7.63(.10) 5.22(.57) 5.17(.61) 4.90(1.07) Ml) 24.5(1.4) 8.05(.ll) 5.77(.90) .72(.15) 24.4(1.2) 8.06(.12) :.86) ------- Table 2 (continued). MEANS AND STANDARD DEVIATIONS (IN PARENTHESES) OF CONDITIONS IN ALL TEST CHAMBERS 00 Test chamber Item 1 2 3 4 5 6 7 8 Test III Fry chamber 1 Guthion^yg/l)-^ . 04 ( . 03)^ . 10 ( . 04) Temp . (C) pH (reading) DO (mg/1) Temp. (C) pH (reading) DO (mg/1) 23.6(.8) 8.22(.08) 7.30(.41) 23.0(.6) 8.23(.08) 7.46(.32) 23.8(.5) 8.17(.04) 6.86(.22) 23.0(.6) 8.16(.05y 6.96(.32) .16(.05) 23.8(.4) 8.15(.05) 6.81(.30) Fry 22.9(.4) 8.16(.05) 7.09(.29) .24(.06) 23. 8(. 8.16(. 6.65(. chamber 23. 0(. 8.15(. 6.99(. 5) 04) 33) 2 6) 05) 34) .33( 23. 9( 8.12( 6.17( 23. 1( 8.13( 6.45( .08) .4) .02) • 41) .6) .07) .51) .51(. 23. 6(. 8.15(. 6.00(. 23.2(1 8.18(. 6.49(. 11) 6) 10) 76) .0) 08) 84) .72(. 23. 6(. 8.14(. 5.87(. 22.4(1 8.15(. 6.06(. 15) 4) 08) 60) .4) 08) 83) a/ j-,Concentration may be high by as much as 50% for the four^lowest concentrations. —/Unknown source of contamination resulted in some Guthion^ in control. — Concentrations measured in adult chambers. ------- were distributed in a random stratified manner to egg chambers in each toxicant concentration and control. Initial numbers of eggs were 90, 70, and 100 in tests I, II, and III, respectively. Eggs were in the middle to late blastula stage when first exposed to the toxicant and more than 99% were fertile. Mortality of eggs and fry was recorded at 24-hr intervals in tests I and II. In test III eggs were checked at 24-hr intervals but mortality of young was recorded only after 22 days to avoid increased stress from handling. At that time two runts or malformed fish were removed from each chamber and fish were randomly thinned to 15 fish. Ten-cm sections of transite pipe were split lengthwise to form a spawning substrate. After 90 days six of these sections were placed in each aquarium, three on each end with an open area in the center. As secondary sexual characteristics developed, males were removed at bi- weekly intervals to keep no more than five per chamber. Spawning sub- strates were checked at 1000 hours each day and if spawning occurred, eggs were counted and a random sample of 50 was placed in the egg basket for determination of survival through hatching unless the basket was already occupied. Two groups of fry from each concentration were reared for 50 days, otherwise all hatched fry were discarded. Fry were fed ground Glencoe trout pellets and live brine shrimp twice a day for the first 30 days. Older fish were fed Oregon Moist frozen pellets twice a day and live brine shrimp once a day. CHEMICAL ANALYSIS Concentrations of Guthion6^ in each adult test chamber were usually analyzed twice per week for the first 5 months of test III and usually once per week thereafter. Samples from either or both fry chambers were analyzed periodically. Since the percentage difference between fry chambers and adult chambers analyzed at the same time ranged from -24% to +21% with a mean percentage difference of iO%, the analyses from adult chambers were used. ------- A 1-liter sample of water from each test chamber was extracted with 10 ml benzene (pesticide grade) and the benzene was then evaporated to 0,5 to 2 ml, depending on the concentration. With each series of samples (§) 1 liter of deionized water was spiked with 1 ml Guthion^ in acetone and this sample was treated the same as all others to determine a recovery efficiency which averaged 99.2 - 6.5%. Samples were analyzed on a Beck- man GC 72-5 gas chromatograph with a 122-cm glass column packed with 50% DC 200 on 80/100 mesh Gas Chrom-Q and a nonradioactive electron capture detector. Standards were prepared from 93% technical grade Guthion^ and were injected alternately with unknowns. Final Guthion*^concentrations were corrected for the recovery efficiency and the technical grade pro- duct. The pH was measured with a meter twice per week in adult chambers and once per week in fry chambers when they were occupied. Temperature was measured with a thermister three times per week in all chambers and dis- solved oxygen was analyzed weekly by the azide modification of the Winkler analysis. 10 ------- SECTION V RESULTS EGG AND FRY SURVIVAL Survival of eggs was probably not directly affected by any concentration /g\ of Guthion** tested. In Test I percentage survival through hatching ® decreased from 90 to 29 to 17% as Guthion concentration increased from 34.5 to 61.4 to 96.9 yg/liter but then survival increased to 76% ® as Guthion concentration increased to 165.9 yg/liter (Table 3). This decreased survival was caused by eggs in these concentrations being /&, enmeshed in a bacterial network that apparently used Guthioir* as an energy source and grew in proportion to the concentration. Dead eggs in chambers with reduced survival were partly hatched as the fry seemed to become enmeshed in the bacteria and could not break free. Since the chorion of these eggs softened, fry may also have been weakened. Sur- vival in the highest concentration was apparently greater because most eggs hatched 1 day sooner, when the bacteria were less dense and before the chorion had softened. At 22 days there was decreased survival of fry in comparison to controls at 7.6 yg/liter in Test I, and at approximately 6.5 yg/liter in Test II (Table 3). By 57 days survival decreased at approximately 1.8 yg/liter (R) Guthion^ in Test II but did not decrease at 0.51 yg/liter in Test III. In Test III Guthion caused no decrease in survival at any life history stage with the possible exception of the highest concentration where fry were lost wheh the chamber overflowed. The MATC in terms of sur- vival was between 0.51 yg/liter in Test III and a maximum of 1.8 yg/liter 11 ------- Table 3. PERCENTAGE SURVIVAL OF FATHEAD MINNOW EGGS AND YOUNG EXPOSED TO VARIOUS CONCENTRATIONS OF GUTHIOt Test chamber Item Guthion^Cug/l) % hatched % survival 22 days Guthion (yg/1) % hatched % survival 22 days 57 days 98 days /**> Guthion^Xyg/l) % hatched % survival 22 days from 22- 60 days 1 0 99 54 0 91 57 54 52 2 7.6 84 43 1.8 92 48 42 35 .O^.IO 86 61 100 84 80 100 3 Test 11.7 97 29 Test 1.9 90 57 34 21 Test .16 82 74 93 4 I 20.5 95 16 a/ II- 2.8 90 43 5 0 III .24 85 82 100 5 34.5 90 1 3.4 87 48 3 0 .33 85 76 100 6 61.4 29 0 - 4.8 93 48 0 0 .51 86 67 93 7 8 96.9 165.9 17 76 0 0 6.5 15.0 63 84 37 35 0 0 0 0 .72 83 c/ v/ _ 88 a/ — Concentration may be high by as much as 50% for the four lowest con- centrations . — Unknown source of contamination resulted in some Guthion in the control. c7 — Tank overflowed and fry were lost. 12 ------- where increased mortality occurred in Test II. GROWTH In test II fish were ewighed twice en masse during the experiment and at termination. Each time the control fish weighed more than those in the lowest concentration, 1.8 yg/liter (Table 4). In test III the first two weighings were of individual fish that were randomly thinned from the experiment. The third weighing was of fish that had remained in the experiment. Individual t-test comparisons indicated no significant dif- ference between the control and any of the treatments (p>.05) at 22 days (Table 4). At 60 days the 0.10 and 0.24 yg/liter treatment levels weighed significantly more than the controls (p<.02) but since the 0.16 yg/liter treatment level was not different from the control, Guthion was probably not the cause of the above differences. After 90 days there was no significant difference (p>.05) between the control and any treatment. Although dissolved oxygen concentrations were lower with increasing Guthioir^ concentrations, these oxygen levels did not seem to affect growth. By 120 days males in some treatments started to defend territories and fish were not weighed to avoid any influence of handling on spawning behavior. At termination of the experiment after 250 days there was no consistent pattern of growth related to Guthioir concentration. There- fore, Guthion5^ affected growth of fathead minnows through 90 to 100 days at essentially the same concentrations that caused mortality, or between 0.72 and 1.8 yg/liter. REPRODUCTION Interpretation of fecundity data was complicated by the variation in sex ratios in test chambers. Although an attempt was made to maintain five males and seven females in each chamber, in some chambers that many males were not produced, in some immature males could not be identified, and in some individuals were lost to accidental death or escapement. 13 ------- Table 4. MEAN WEIGHT OF FATHEAD MINNOWS AT VARIOUS TIME INTERVALS AFTER SPAWNING (NUMBER OF FISH WEIGHED IN PARENTHESES) Item Test chamber 12 3 4 5 6 7 8 Guthion® (ug/1) Weight (mg) 57 days 85 days 98 days Guthion® (yg/1) Weight (mg) 1.8 Test II 1.9 2.8 3.4 4.8 162(33) 111(29) 55(21) 72(3) 20(2) 320(33) 250(24) 230(14) 380(33) 350(24) 300(13) 0.4 0.10 Test III 0.16 0.24 0.33 0.51 6.5 0.72 a/ j-.Fish randomly thinned from experiment. —.Fish remaining in experiment. — Termination. 15.0 22 60 90 250 days^ days- days— days— - male female 24(8) 179(15) 580(15) 2834(3) 1434(9) 24(27) 242(15) 580(15) 3047(5) 1594(7) 28(25) 216(13) 640(15) 3455(5) 1458(5) 27(19) 231(15) 660(14) 3051(5) 1353(5) 23(21) 260(15) 600(15) 2628(5) 1558(5) 21(11) 165(13) 600(13) 2965(6) 1360(6) _ _ 720(15) 3136(5) 912(1) ------- Spawning occurred after 121 days when the day length reached 12 hr and 45 min. Spawning ceased after 230 days when the day length was reduced to 12 hr. Intensive spawning occurred first at 0.33 yg/liter GuthionY followed 12 to 14 days later by spawning at 0.24, 0.16 and 0.10 yg/liter. Intensive spawning by the control fish occurred 29 days later and 56 days later in 0.51 and 0.72 yg/liter Guthiort-' although at 0.51 yg/liter spawning occurred infrequently during the test. The number of spawnings and eggs were greatest in the control chamber (Table 5). This result may have been caused by the highest female to male sex ratio throughout the test. The more significant measurements in terms of GuthiorHeffect were the number of eggs per spawning and per female since these nor- malize the dissimilar sex ratios. On this basis Guthioff-'affected fecundity between 0.33 yg/liter which was similar to controls and 0.51 yg/liter where there was about a 50% reduction in eggs per spawning and per female (Table 5). The decreased oxygen concentrations with increas- ing Guthion-'was apparently not responsible for the effect on fecundity since the oxygen concentration at 0.33 and 0.51 yg/liter GuthioA-^was similar. Table 5. FECUNDITY OF FATHEAD MINNOWS EXPOSED TO VARIOUS CONCENTRATIONS OF GUTHION5^ GuthiorP yg/l 0.04 0.10 0.16 0.24 0.33 0.51 0.72 Number of spawnings 88 54 57 49 52 21 27 Number of eggs 16401 t 8541 9567 7091 8590 1791 2408 Number of eggs per spawning 186 158 168 145 165 85 89 Number of eggs per female^/ 1691 1220 1611 1239 1718 256 782 a/ — Computed by intervals between thinnings. 15 ------- SECOND GENERATION Random samples of 50 eggs were taken from 12 spawnings from each test chamber except the second highest where samples from seven spawnings were taken. These eggs were hatched in the oscillating egg baskets. As with the first generation eggs, there was no effect of Guthiott^ at the tested concentrations (Table 6). Table 6. EFFECT OF ON SURVIVAL AND GROWTH OF SECOND GENERATION EGGS AND FRY GuthionS/ concentration, yg/1 Item % survival - eggs % survival - fry- Is t group 2nd group Mean weight (mg)- 1st group 2nd group 0.04 86.2 79 69 94 143 0.10 84.2 97 89 130 102 0.16 90.9 100 88 94 135 0.24 86.2 97 89 126 85 0.33 82.3 89 100 115 144 0.51 0.72 86.4 90.2 - -1 78 100 78 128 222 80 a/ — After 50 days from hatching. — Some fry lost due to overflow. Survival or growth of fry was not consistently affected by the tested Guthion*^ concentrations (Table 6). Survival was poorest among controls. Considerable differences in growth occurred between test chambers but these differences were not related to Guthiori-' concentrations (Table 6). 16 ------- SECTION VI DISCUSSION The lowest concentration of Guthion^ that had an adverse effect on fat- head minnows was 0.51 yg/liter where fecundity was much reduced (Table 5). An estimation of the maximum acceptable toxicant concentration based on this parameter is between 0.33 and 0.51 yg/liter Guthionr Effects on survival and growth during early life history stages occurred at slightly higher concentrations (Tables 3 and 4). Although no effect occurred in test III at the highest concentration (0.72 yg/liter) growth and survival were reduced at the lowest concentration (1.8 yg/liter) in test II. The 96-hr LC50 for 11-week-old fathead minnows tested in a relatively soft water was 1900 yg/liter (Part I of this report). An application factor based on this acutely toxic concentration and the MATC in the present study is between .00017 and .00027. Although this application factor is extremely small compared to factors found for many other toxicants (U.S. Environmental Protection Agency, 1973), it would prob- ably be even smaller if the acute tests were conducted in hard water. Henderson, Pickering and Tarzwell (1960) found that the 96-hr LC50 of Guthioir-^to fathead minnows was approximately 25 to 40% less in soft water than in hard (20 ing/liter vs. 400 mg/liter total hardness). This differential in hardness was more extreme than the acute vs. chronic test in the present study, but if the application factor was based on an acute test conducted in hard water, the implication is that the factor would be even smaller. 17 ------- SECTION VII REFERENCES DeFoe, D.L. 1975. Multichannel Toxicant Injection System for Flow- through Bioassays. J. Fish. Res. Board Can. 32;544-546. Henderson, C., Q.H. Pickering, and C.M. Tarzwell. 1960. Toxicity of Organic Phosphorus and Chlorinated Hydrocarbon Insecticides to Fish. In: Biological Problems in Water Pollution. Trans. 1959 Seminar. Cincinnati, R.A. Taft Sanit. Eng. Center, p. 76-92. Katz, M. 1961. Acute Toxicity of Some Organic Insecticides to Three Species of Salmonids and to the Three-spine Stickleback. Trans. Amer. Fish. Soc. 90(3):264-268. Macek, K.J., and W.A. McAllister. 1970. Insecticide Susceptibility of Some Common Fish Family Representatives. Trans. Amer. Fish. Soc. 99: 20-27. Meyer, F.P. 1965. The Experimental Use of Guthion as a Selective Fish Eradicator. Trans. Amer. Fish. Soc. 94(3);203-209. Mount, D.I., and W.A. Brungs. 1967. A Simplified Dosing Apparatus for Fish Toxicology Studies. Water Res. 1(1):21-29. Mount, D.I., and C.E. Stephan. 1967. A Method for Establishing Accept- able Toxicant Limits for Fish - Malathion and the Butoxyethanol Ester of 2,4-D. Trans. Amer. Fish. Soc. 96:185-193. 18 ------- Pickering, Q.H., C. Henderson, and A.F. Lemke. 1962. The Toxicity of Organic Phosphorus Insecticides to Different Species of Warm Water Fishes. Trans. Amer. Fish. Soc. 91(2);175-184. U.S. Environmental Protection Agency. 1973. Water Quality Criteria 1972. Ecol. Res. Ser. EPA-R3-73-033. 594 p. 19 ------- SECTION VIII PUBLICATIONS Adelman, I.R., L.L. Smith, Jr., and G.D. Siesennop. Chronic Toxicity of Guthion to the Fathead Minnow (Pimephales promelas Rafinesque). Bulletin of Environmental Contamination and Toxicology (submitted). 20 ------- SECTION IX GLOSSARY Adult - A fish older than 4 months. Application Factor - The ratio of the MATC to the 96-hr LC50. Bioassay - A toxicity test; the estimation of the strength of a poison by its effect on a living organism. Bioassay, acute - A toxicity test of short duration, usually less than 3 weeks. Bioassay, chronic - A toxicity test of long duration, usually 1 month or longer. Bioassay, flow-through - A toxicity test where the toxicant and diluent water are continuously replaced by fresh material. Biastula - Early embryonic stage where cellular division has resulted in cells arranged as a hollow spherical body. Chorion - The outer membraneous covering of a fish egg. Fry - A fish younger than about 50 days. LC50 - The concentration of poison that will kill 50% of the test organisms at a specified time. 21 ------- Maximum Acceptable Toxicant Concentration (MATC) - A concentration of toxicant determined from a chronic bioassay and considered an estimation of the level at which no harm will occur in the natural environment. It is bracketed by the highest level of no effect and the lowest level of effect in the bioassay. 22 ------- TECHNICAL REPORT DATA (Please read Instructions on the reverse before completing) 1. REPORT NO. EPA-600/3-76-061b 3. RECIPIENT'S ACCESSION>NO. 4. TITLE AND SUBTITLE STANDARD TEST FISH DEVELOPMENT, PART II 5. REPORT DATE July 1976 (Issuing Date) 6. PERFORMING ORGANIZATION CODE 7. AUTHOR(S) Ira R. Adelman and Lloyd L. Smith, Jr. 8. PERFORMING ORGANIZATION REPORT NO 9. PERFORMING ORGANIZATION NAME AND ADDRESS Department of Entomology, Fisheries, and Wildlife University of Minnesota St. Paul, Minnesota 55108 10. PROGRAM ELEMENT NO. 1BA608 11. CONTRACT/GRANT NO. Grant R800940 12. SPONSORING AGENCY NAME AND ADDRESS U.S. Environmental Protection Agency Office of Research and Development Environmental Research Laboratory Duluth, Minnesota 55804 13. TYPE OF REPORT AND PERIOD COVERED T7TNAL 14. SPONSORING AGENCY CODE EPA-ORD 15. SUPPLEMENTARY NOTES See Part I, EPA-600/3-76-061a 16. ABSTRACT Three chronic bioassays of GuthiorTwere conducted with fathead minnows. All tests were begun with eggs, and the longest lasted 20 days after termination of spawning, a total of 250 days. Parameters measured were survival, growth, fecundity, and growth and survival of second generation fry and eggs. The most sensitive criteria for effect of the toxicant was fecundity. The maximum ^ acceptable toxicant concentration was between 0.33 and 0.51 yg/liter Guthion^ and the application factor between .00017 and .00027. 17. KEY WORDS AND DOCUMENT ANALYSIS DESCRIPTORS b.lDENTIFIERS/OPEN ENDED TERMS C. COS AT I Field/Group Bioassay Criteria Fecundity Growth Minnows Water quality Survival Guthion Chronic toxicity Bioassay survival Application factor Maximum acceptable toxicant concentration 06C 06F 06S 18. DISTRIBUTION STATEMENT RELEASE TO PUBLIC 19. SECURITY CLASS (ThisReport) UNCLASSIFIED 21. NO. OF PAGES 31 20. SECURITY CLASS (Thispage) UNCLASSIFIED 22. PRICE EPA Form 2220-1 (9-73) 23 •OUSGPO: 1976 — 657-695/5461 Region 5-11 ------- |