| V5 ji fishes INVESTIGATIONS IN FISH CONTROL 95. Deposition and Persistence of Rotenone in Shallow Ponds During Cold and Warm Seasons aR AEA) 9, UNITED STATES DEPARTMENT OF THE INTERIOR FISH AND WILDLIFE SERVICE Investigations in Fish Control, published by the Fish and Wildlife Service, include reports on the results of work at the National Fishery Research Center at La Crosse, Wisconsin, and reports of other studies related to that work. Though each report is regarded as a separate publication, several may be issued under a single cover, for economy. See inside back cover for list of current issues. Copies of this publication may be obtained from the Publications Unit, U.S. Fish and Wildlife Service, Washington, DC 20240, or may be purchased from the National Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Library of Congress Cataloging-in-Publication Data Gilderhus, P. A. (Philip A.) Deposition and persistence of rotenone in shallow ponds during cold and warm seasons. (Investigations in fish control ; 95) 1. Rotenone—Environmental aspects. 2. Pond fauna—Effect of insecticides on. 3. Pond ecology. I. Dawson, Verdel K. II. Allen, J. K. John L.) If. Title. IV. Series. SH157.7.158 No. 95 639.9'77 s [639.3'11] 88-600290 [QH545.P4] INVESTIGATIONS IN FISH CONTROL 95. Deposition and Persistence of Rotenone in Shallow Ponds During Cold and Warm Seasons By P. A. Gilderhus, V. K. Dawson, and J. L. Allen UNITED STATES DEPARTMENT OF THE INTERIOR FISH AND WILDLIFE SERVICE Washington, D.C. © 1988 ‘alta 1 1 aa IY me | =~» atom tow toe Publlcntiay tina; Uae ere Lae ayy 22 jena Pirate fe Narhinal sactonormly mantinarnatit Deposition and Persistence of Rotenone in Shallow Ponds During Cold and Warm Seasons P. A. Gilderhus, V. K. Dawson, and J. L. Allen U.S. Fish and Wildlife Service National Fisheries Research Center P.O. Box 818 La Crosse, Wisconsin 54602 Abstract As part of the requirements for the continued registration of rotenone, data were developed on deposition and persistence of the chemical in aquatic ecosystems in cold and warm seasons. After treatment of two ponds, one in November (0-5° C) and one in July (23-27° C), with 5 uL/L of Noxfish (0.250 mg/L of rotenone), we collected samples of water, bottom sediments, invertebrates, and fish and analyzed them for rotenone by high performance liquid chromatography. Decomposi- tion of rotenone in water followed a first-order decay curve; half-life was 10.3 days in cold water and 0.94 days in warm water. In cold water, residues in bottom sediments gradually increased to a peak of 0.1 yg/g after 14 days and then declined to <0.025 yg/g (the limit of detection) after 64 days; in warm water, residues in sediments fell below the detection limit within 24 h. In fresh- water mussels and crayfish, residues gradually increased for 1 week in cold water and 1 day in warm water and then slowly decreased. Residues in fish varied with species and water temperatures; concentrations were higher in fish tissues than in the water on corresponding days. Rotenone has been used as a fish toxicant in the United States for about 50 years, and is currently the most wide- ly used piscicide in the United States (Schnick 1974). Since rotenone was first registered with the U.S. Envi- ronmental Protection Agency (EPA) in 1962, the require- ments for registration of pesticides have increased and have been revised several times. The guidelines now require more extensive documentation of the safety, efficacy, and persistence of chemicals in the environment. All previously registered pesticides are required to con- form to the revised standards when their labels are reviewed. Preliminary studies have indicated that the per- sistence of rotenone is brief in warm water and much longer in cold water (Gilderhus et al. 1986). Such infor- mation was helpful in developing adequate sampling schedules for more comprehensive studies. We describe research performed in 1983 and 1984 on persistence of rotenone in the environment to meet cur- rent EPA requirements for continued registration. The objective of the studies was to determine where rotenone is deposited in the environment, in what amounts, and how long it stays there during different seasons. The studies were conducted in fall and summer in shallow ponds to determine the distribution and persistence of rotenone in water, bottom sediments, invertebrates, and fish under cold and warm conditions. For convenience, we refer to the pond treated in November as the ‘‘coldwater pond”’ and the one treated in July as the “‘warmwater pond.”’ These designations do not imply any marked difference in thermal regimen or biota between the ponds. Materials and Methods Study Sites The coldwater pond was at the Genoa (Wisconsin) National Fish Hatchery and the warmwater pond at the La Crosse National Fisheries Research Center; their physical, chemical, and sediment characteristics are shown in Table 1. Use of two different ponds was neces- sary because we did not have access to the hatchery pond 1 Table 1. Characteristics of ponds used in studies on the persistence of rotenone in the environment. Coldwater Warmwater Characteristics pond pond Pond and water Approximate dimensions (m) IS 33027 xs Volume (m3) 3,526 185 Surface area (ha) 0.46 0.02 Mean depth (m) 0.78 0.86 pH 8.62 8.35 Turbidity (NTU)? 3.43 2.65 Total hardness (mg/L as CaCO3) 284 113 Total alkalinity (mg/L;as CaCO3) 253 92 Sediment Sand (%) 91 93 Silt (%) 7 4 Clay (%) 2 3 Organic content (%) 2.4 2.3 Moisture content (%) 41.1 29.0 Cation exchange capacity 14.45 13.04 (meq/100 g) Density (g/cm?) 1.30 1.51 pH 8.2 8.1 4Nephelometric turbidity units. during summer and the outlet structure in the pond at the Research Center would not withstand the pressure from thick ice. At the time of treatment of the coldwater pond on 14 November 1983, the water temperature was 5° C and remained at 4 to 5° C until 28 November, when the pond became covered with ice about 2.5 cm thick. Water temperature directly under the ice was 0° C by 5 Decem- ber; ice thickness increased to 25 cm by 27 December and then decreased to 15 cm by 29 February. The warm- water pond was treated on 30 July 1984; water temper- atures ranged from 23 to 27° C as long as 7 days after treatment, when rotenone was no longer detected in water or soil. Treatments Each pond was treated with Noxfish, a commercial for- mulation containing 5% rotenone, to produce a concen- tration of 0.250 mg/L of rotenone (the maximum allowed by the product label). The formulated chemical was diluted 8:1 with pond water before it was applied. In the coldwater pond, we used a boat with a venturi-type boat bailer on a 6-horsepower outboard motor (a small, shallow area at one end of the pond was treated with a hand- pumped sprayer). We treated the shallow end of the warm- water pond with the hand-pumped sprayer and the rest of the pond by using the boat bailer with the motor mounted on the water-control structure at the deep end of the pond. Sampling Samples of the various components in the aquatic en- vironment (water, bottom soil, and experimental animals) were collected in accordance with Pesticide Assessment Guidelines, Subdivision N—Chemistry: Environmental Fate (EPA 1982). Sampling schedules were based on data from the preliminary studies on the persistence of rote- none in water (Gilderhus et al. 1986). Water samples were collected in amber glass jugs sub- merged just below the surface of the water. Samples were collected along the midline of the coldwater pond at points 30, 60, and 90 m from one end, either from a boat or through holes in the ice made with an ice spud or gasoline- powered auger. Water samples from the warmwater pond were also collected along the midline, at points 3 m from either end and at the center of the pond. All water samples were extracted within 2 h after collection. Samples of bottom soil were collected with a core sampler 5 cm in diameter, similar to that described by Swanson (1978), at the same locations where the water samples were collected. Each sediment sample was a com- posite of three cores taken within a radius of 0.5 m. Cores were collected by forcing the sampler 10-15 cm into the bottom, and then releasing the core into an enameled pan; the top 5 cm was then cut off and placed into a plastic container with a snap-on lid, along with the top 5 cm of each of the two other cores in the sample. Samples were placed in a freezer at —10° C within 2 h after collection. Particle sizes of sediments were determined according to standardized procedures for sieving and hydrometer classification (ASTM 1979). Soil classifications were pro- vided by the University of Wisconsin Soil and Forage Laboratory, Marshfield, Wisconsin. Freshwater mussels and crayfish were used as repre- sentative invertebrates because they are easy to sample and are large enough to provide adequate tissue samples (10 g) for rotenone analysis. The mussels (Lampsilis sp.) were collected from Lake Onalaska near La Crosse, Wisconsin, and the crayfish (Orconectes sp.) were ob- tained from the Wisconsin Department of Natural Re- sources. Mussels were placed in a woven wire cage resting on the pond bottom, at a single location in each pond; sides of the cage extended above the water. A 10- to 15-cm layer of sand in the bottom of the cages provided a sub- strate in which the mussels could burrow. Mussels were placed in the coldwater pond 1 week before treatment and the warmwater pond 2 weeks before treatment. At each sampling, six mussels were removed from the cage, rinsed in clean water for 30 s, and packaged as three replicates of two mussels each. The soft tissues were analyzed for rotenone and the shells were discarded. Crayfish were placed in a single floating cage 1 week before treatment in each pond. At each sampling, three groups of three large crayfish were taken from the cold- water pond and three groups of six smaller crayfish from the warmwater pond (to ensure samples of adequate size for analysis). To aid handling and packaging, we killed crayfish immediately after collection by inserting a sharp blade into the brain. Whole crayfish were ground and extracted for rotenone analysis. Invertebrates were wrapped in aluminum foil, placed in a polyethylene bag, and sealed. Samples were frozen within 2 h after collection and stored at —10° C until analyzed. Fish resident in the ponds (common carp, Cyprinus car- pio, and largemouth bass, Micropterus salmoides, in the coldwater pond; shortnose gar, Lepisosteus platostomus, in the warmwater pond) were sampled after they were killed by rotenone on the day of treatment. Three com- mon carp, three largemouth bass, and two shortnose gar were sampled and processed individually; the fillets and offal were packaged separately in foil, placed together in a plastic bag, and sealed. Since resident fish killed by rotenone are not recom- mended for consumption as food, emphasis was placed on uptake and elimination of rotenone by fish that were stocked after the water was no longer toxic. Beginning on day 7 in the coldwater pond and day 1 in the warm- water pond, we placed fathead minnows, Pimephales pro- melas, in floating cages to monitor the toxicity of the water. When 9 of 10 fish survived for 24 h (on days 30 and 4 after treatment in coldwater and warmwater ponds, respectively) we considered the water to be safe for restocking. We chose to use an ictalurid and a centrarchid in each pond (different species of these families were avaliable to us when the two tests were conducted). Chan- nel catfish, Ictalurus punctatus, and largemouth bass (mean total lengths, 37 and 23 cm, respectively) were placed in cages in the coldwater pond on day 30 after treat- ment. Black bullheads, Ictalurus melas, and bluegills, Lepomis macrochirus, averaging 25 and 13 cm in length, respectively, were placed in the warmwater pond on day 7 after treatment. Fish cages rested on the bottom and ex- tended above the water surface. For the larger fish (chan- nel catfish, black bullheads, and largemouth bass), three fish were taken at each sampling; the fillets and offal were packaged and analyzed separately for each fish. Bluegills were sampled as three groups of four fish on each sam- pling day; fillets and offal from each group of four were pooled and packaged separately. Packaging and freezing procedures for fish were the same as those used for invertebrates. Sample Analyses Water samples were acidified to pH 5 with an acetate buffer and extracted with a disposable Baker C;g chroma- tography column. The extracted rotenone was eluted from the column with 2 mL of methanol and analyzed by high performance liquid chromatography (HPLC) as described by Dawson et al. (1983). A Waters model M-45 HPLC with a Waters 15 cm X 3.9 mm Nova-Pak Cg reverse- phase column and UV detector (295 nm) was used for the analyses. The mobile phase consisted of methanol: water (70:30 v/v) at a flow rate of 1 mL/min. The lower limit of detection for water was 0.002 mg/L. Sediment samples were extracted with methanol on a Sorval mixer, centrifuged, and filtered on Gelman type A/E glass fiber filters. The extracts were acidified, par- titioned into hexane, and transferred to a silica gel col- umn. The samples were eluted from the silica gel with benzene:acetone (97:3), exchanged into methanol, and analyzed by HPLC by the modified method of Bowman et al. (1978). The detection limit for rotenone in sediments was 0.025 ug/g. Fish fillets, fish offal, crayfish (whole body), and fresh- water mussels (without shell) were homogenized in a blender with dry ice according to the procedure of Ben- ville and Tindle (1970). Homogenates were mixed with sodium sulfate and column-extracted with ethyl ether (Hesselberg and Johnson 1972). We separated extracts from lipids by gel permeation chromatography (GPC) with SX-3 biobeads and methylene chloride:cyclohexane (1:1 v/v). Further cleanup was obtained by silica gel chroma- tography in the same way as described for the sediment samples, and was followed by HPLC analysis. The lower limit of detection for rotenone in tissues was 0.005 yg/g. Quality assurance for the analytical portion of the study was provided by systematic analysis of blanks, replicates, and spiked controls. Results Water Rotenone degraded much more slowly in the coldwater pond than in the warmwater pond (Table 2). In the cold- water pond, the mean concentration of rotenone in the initial samples taken 3 h after treatment was 0.229 mg/L; the concentration declined gradually but steadily over time; and residues dropped to <0.002 mg/L (the limit of detection) after 57 days (Table 2). In the warmwater pond, the mean concentration of rotenone at 3 h was 0.180 mg/L; the concentration declined by more than 50% in the first 12 h; and residues fell to the 0.002-mg/L detection limit within 4 days (Table 2). The rate of loss of rotenone from water followed a first-order decay curve; the half-life was 10.3 days in the coldwater pond and 0.94 day (22.5 h) in the warmwater pond (Fig. 1). Table 2. Mean (N = 3) concentrations of rotenone (standard errors in parentheses) in samples collected from a coldwater and a warmwater pond treated with 5 L/L of Noxfish (0.250 mg/L of rotenone). Pond and Water posttreatment time (mg/L) Coldwater* Hours 3-6? 0.229 (0.010) Days 1 0.186 (0.003) 3 0.164 (0.004) 7 0.090 (0.004) 14 0.062 (0.003) 21 0.030 (0.005) 28 0.020 (0.001) 36 0.020 (0.002) 43 0.012 (0.000) 50 0.006 (0.000) 57 0.0024 64 =a 78 — Warmwater® Hours 3 0.180 (0.002) 6 0.154 (0.002) 12 0.110 (0.005) 18 0.097 (0.010) Days 1 0.089 (0.001) Sediments Crayfish Mussels (ug/g) (ug/g) (ug/g) 0.029 0.188 0.068 (0.004) (0.009) (0.0000) 0.034 0.395 0.066 (0.004) (0.046) (0.032) 0.058 0.340 0.174 (0.008) (0.014) (0.048) 0.075 0.235 0.723 (0.014) (0.033) (0.099) 0.100 0.200 0.696 (0.025) (0.065) (0.033) 0.075 0.057 0.382 (0.014) (0.008)° (0.145) 0.054 — 0.230 (0.011) — (0.051)° 0.042 = ae (0.008) — = 0.033 — = (0.008) — = <0.025¢ — _ <0.025¢ — = 0.075 0.088 0.305 (0.014) (0.050) (0.077) 0.033 — == (0.008) — = <0.025¢ 0.076 1.060 — (0.00) (0.427)° Table 2. Continued. Pond and Water posttreatment time (mg/L) 1.5 0.061 (0.001) 2 0.040 (0.006) 3 0.020 (0.000) 4 0.0024 7 = 4Water temperature at time of treatment, 5° C. Water and sediments, 3 h; crayfish and mussels, 6 h. Mussels (ug/g) Sediments (ug/g) Crayfish (ug/g) = 0.058 — = (0.008) — = 0.045 — = (0.022) — — 0.019 — = (0.017) — <0.0254 <0.005¢ = °Last sample because no live animals remained at the next sampling date. 4] imit of detection. ©Water temperature at time of treatment, 24° C. Sediments Bottom sediments in the two ponds had similar physical characteristics (Table 1). Consequently, their adsorptive capacity for rotenone was probably similar because this capacity is closely related to particle size and organic con- tent (Dawson et al. 1986). Residues of rotenone in bottom sediments in the coldwater pond peaked at 0.100 yg/g after 14 days and then declined to <0.025 yg/g (limit of detec- tion) after 64 days (Table 2). Accumulation and elimina- tion were much faster in the warmwater pond; the concentration in the sediments peaked at 0.075 ug/g after 6h and dropped to <0.025 yg/g after 24 h (Table 2). The peak concentration in the sediments was higher than that oo A x cold water o warm water Concentration (mg/L) Days Fig. 1. Disappearance of rotenone from pond waters treated with 5 wL/L of Noxfish (0.250 mg/L of rotenone). in water in the coldwater pond but less than half that in water in the warmwater pond. Crayfish Crayfish in the coldwater pond accumulated concen- trations of rotenone 1.58 times the treatment concentra- tion by 1 day after treatment; concentrations then declined steadily from day 1 to day 21 (Table 2). All caged cray- fish were dead by day 28, possibly as a result of the long submersion in cold water during a season when they would normally be hibernating. However, delayed mortality due to rotenone toxicity is also a possibility. In warm water, residues of rotenone in crayfish peaked at 0.088 ug/g (35% of the treatment concentration) 6 h after treatment (Table 2). The concentration declined rapidly to the limit of detection (0.005 g/g) by day 7. The decline of residues in crayfish closely paralleled that in water in the warmwater pond. Mussels The concentrations of rotenone accumulated were higher in mussels than in crayfish. In the coldwater pond, residues peaked at 0.723 ug/g (2.88 times the treatment concentration) at 7 days after treatment, and declined to 0.230 ug/g (Table 2) on day 28. In the warmwater pond, tissue concentrations reached 1.060 ug/g (4.24 times the treatment concentration) 1 day after treatment (Table 2). Because all of the mussels had died, no further sampling was possible. Since the caged mussels, which had been in the pond for 2 weeks, were alive on the day of treat- ment and dead the next day, we assumed that the mor- talities were due to the toxic effects of rotenone. Fish Fish in the ponds were dead within 2 to 4 h after treat- ment. Residues of rotenone in fillets from these fish were generally commensurate with the concentrations in the water. In the coldwater pond, common carp accumulated the highest concentrations (0.329 + 0.046 ug/g in fillets) and largemouth bass the lowest (0.171 + 0.008 g/g in fillets). In shortnose gar in the warmwater pond, residues were 0.202 + 0.116 yg/g in fillets. Concentrations in the offal were about double those in fillets of each species. Fish stocked in the ponds after the rotenone had declined to nontoxic levels accumulated rotenone residues that were higher than those in the invertebrates that were in the pond on the day of treatment. In cold water, both channel cat- fish and largemouth bass accumulated rotenone residues in excess of 20 times the level in the water when the fish were stocked (Table 3). Peak concentrations in the fillets and offal of channel catfish were about equal (Table 3). In largemouth bass, the peak in the fillets was nearly 3 times that in the offal. In the warmwater pond, black bullheads concentrated rotenone up to 76 times the con- centration in the water (0.002 mg/L) when the fish were stocked into the pond (Table 3). Their uptake and elimina- Table 3. Concentrations* of rotenone in channel catfish and largemouth bass stocked in a coldwater pond 30 days after it was treated with 5 L/L of Noxfish (0.250 mg/L of rotenone) and in black bullheads and bluegills stocked in a warmwater pond 7 days after a similar treatment. Channel catfish Largemouth bass Days after stocking Fillet Offal Fillet Offal 1 0.123 0.228 0.082 0.166 (0.018) (0.050) (0.016) (0.013) 3 0.360 0.351 1.225 0.440 (0.025) (0.016) (0.037) (0.027) 6 0.410 0.417 0.502 0.263 (0.034) (0.077) (0.20) (0.083) 13 0.131 0.342 — — (0.011) (0.068) — — 20 0.178 0.325 — — (0.026) (0.038)? _ — Black bullheads Bluegills Fillet Offal Fillet Offal 1 0.005° 0.032 0.064 0.067 (0.000) (0.030) (0.005) (0.005) 3 0.005° 0.054 <0.005° 0.087 (0.000) (0.004) — (0.008) 7/ 0.045 0.062 <0.005° 0.078 (0.042) (0.003) — (0.006) 14 0.153 0.096 <0.005° <0.005° (0.036) (0.016) — — 21 0.083 0.026 <0.00S° <0.005° (0.004) (0.016) _— = 29 0.060 0.018 — = (0.004) (0.016) — — 35 <0.005° 0.005° — — *Mean of three samples—standard errors in parentheses. Last sample because no live animals remained at the next sampling date. ©Limit of detection. tion of rotenone spanned 5 weeks. In bluegills, the up- take and elimination of rotenone were rapid, peaking on day 1 and declining to below the limit of detection by day 3 (Table 3). Discussion The length of time that rotenone persisted in the water and the time that it became nontoxic to fathead minnows were about as expected. The degradation of rotenone in aquatic environments is influenced by many environmental factors that are more or less active, depending on the season. Water temperature was a major factor in our study, as in other published studies. The influence of temperature has been expressed in formulas developed by Post (1958) as well as by Engstrom-Heg and Colesante (1979), who also emphasized the effect of sunlight. Our data are in general agreement with those of Post (1958) and Engstrom-Heg and Colesante (1979). Post’s formulas would have predicted persistence of about 46 days in our coldwater pond and 6 days in our warmwater pond. The formulas of Engstrom-Heg and Colesante (1979) would have predicted that the rotenone-treated water would re- main toxic to fish somewhat longer (69 days in our cold- water pond and 7 days in our warmwater pond). Various other factors are known to significantly affect the toxicity of rotenone (Gilderhus 1982). Aquatic plants and suspended clay presumably adsorb and absorb the chemical. Phytoplankton, zooplankton, and bacteria are also likely to influence the rate at which rotenone dis- appears from water and are likely to be most active in warm water. The generally higher concentrations of rotenone in sediments and organisms in the coldwater pond were probably due to the much longer time that they were exposed to detectable concentrations of rotenone in the water. All of the studies on rotenone persistence, including the present one, were based on limited data and thus cannot be considered definitive. To predict how long the toxicity of rotenone will persist, fishery workers should consider the entire body of relevant literature. Although published studies may help to estimate rotenone persistence, on-site tests with fish are always advisable for the final determination of when the treated water is safe for restocking. The rapid accumulation of rotenone in fish tissues in- dicates a high uptake efficiency. The dynamics of chemical movement across the gills of fish were described by McKim et al. (1985), whose studies indicated that chemicals with octanol-water partition coefficients (log P) between 3 and 6 have the highest uptake in fish. Gingerich and Rach (1985) reported the log P for rotenone to be 4.26, placing it in the category of chemicals most easily transported across the gills. Those studies also showed that rotenone tended to concentrate in the viscera and that elimination from the viscera was relatively rapid. Rapid uptake of rotenone was also apparent in our study, in which the concentrations accumulated by fish were much higher than those in the water at the time the fish were stocked. References ASTM (American Society for Testing and Materials). 1979. Standard method for particle-size analysis of soils. Pages 112-122 in Annual book of ASTM standards, Philadelphia, Pa. Benville, P. E., Jr., and R. C. Tindle. 1970. Dry ice homog- enization procedure for fish samples in pesticide residue analysis. Agric. Food Chem. 18:948-949. Bowman, M. C., C. L. Holden, and L. I. Bone. 1978. High pressure liquid chromatographic determination of rotenone and degradation products in animal chow and tissues. J. Assoc. Off. Anal. Chem. 61:1445-1455. Dawson, V. K., P. D. Harmon, D. P. Schultz, and J. L. Allen. 1983. Rapid method for measuring rotenone in water at piscicidal concentrations. Trans. Am. Fish. Soc. 112:725-727. Dawson, V. K., D. A. Johnson, and J. L. Allen. 1986. Loss of lampricides by adsorption on bottom sediments. Can. J. Fish. Aquat. Sci. 43:1515-1520. Engstrom-Heg, R., and R. T. Colesante. 1979. Predicting rotenone degradation rates in lakes and ponds. N.Y. Fish Game J. 26:22-36. Gilderhus, P. A. 1982. Effects of an aquatic plant and suspended clay on the activity of fish toxicants. N. Am. J. Fish. Manage. 2:301-306. Gilderhus, P. A., J. L. Allen, and V. K. Dawson. 1986. Per- sistence of rotenone in pond water at different temperatures. N. Am. J. Fish. Manage. 6:129-130. Gingerich, W. H., and J. J. Rach. 1985. Uptake, biotrans- formation, and elimination of rotenone by bluegills (Lepomis macrochirus). Aquat. Toxicol. 6:179-196. Hesselberg, R. J., and J. L. Johnson. 1972. Column extraction of pesticides from fish, fish food and mud. Bull. Environ. Contam. Toxicol. 7:115-120. McKim, J., P. Schmieder, and G. Veith. 1985. Absorption dynamics of organic chemical transport across trout gills as related to octanol—water partition coefficient. Toxicol. Appl. Pharmacol. 77:1-10. Post, G. 1958. Time versus water temperature in rotenone dissipation. Proc. West. Assoc. Game Fish Comm. 38:279-284. Schnick, R. A. 1974. A review of the literature on the use of rotenone in fisheries. U.S. Fish and Wildlife Service, Fish Control Laboratory, La Crosse, Wis. NTIS No. PB-235 454. 130 pp. Swanson, G. A. 1978. A simple lightweight core sampler for quantitating waterfowl foods. J. Wildl. Manage. 42:426-428. U.S. Environmental Protection Agency. 1982. Pesticide assess- ment guidelines, subdivision N. Chemistry; environmental fate. U.S. Department of Commerce, National Technical In- formation Service, Springfield, Va. PB83-153973. 108 pp. a ect iis SE sein att 4 a Goede che ; 2 eaters _— women att Vet * jy oem aoc iagertia he ili teen * “ Jacvuevit apr LTC ae A | ee ry sstirantio's 2G Pale ie ARMAS TT ee ate ee fvuiy et ie ‘ Cle = ws pe ti! ine wet EL a ae f ntfa! “wi ¥,! seal) ee yt ARIE PRA awn? 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UOTTsoduioseq *Aydeisojewo1y9 prnbiy soueULOy -1od ysiy Aq auous}01 10} wou) pezATeue pur Ysij pue ‘soyeIqoVJOAUT ‘S}USUUTpas Woo ‘138M JO sordures pojoaT[oo am ‘(QUOUD}OI JO J/SUI YSZ'0) YSIXON Jo 7/1" ¢ wim spuod om} Jo JuoUnLAN Joy ‘suOseas ULTeM pur Pjod UT suIa}sAso59 onenbe ut yeorureys oy) Jo douajstsied pue uontsodep uo pedojsasp o10M BJep ‘AUOUD}OI JO UONeINSIZ91 ponuNUOS oy) JO} syuoWIoIINbeI oy) Jo Wed sy "dd 1 °¢6 Jo41u0D Ysiy ‘ISAAUT ‘AIO “TPIEAA USI “S'N “suOsBag ULIBAA PUE P[OD SUING spuog MOT[BYG UI BUOUA}0Y JO adUa}SISIOg PUB uontsodad “8861 “UITy “I UYos pue ‘uosmeEq “yf [9PI9A “Vy dimyd ‘snyrep[iH _ leis ' Ss | eee aes Males Ses mes rr erage Cv ami il be es .~ mes Tee wa? mt eda F stv a a rh) yew ia) : sTiatiee Let GE CAG POY heecilot wu 6 ib = a alae visas Se Bee dE Ol Pears set ate UYWo ost fr nbereash sill rion Sa¥ apbeciiins ni mchaest ae ew f ihe ih wilenterry Mulvey: ae hqerss: hoe Catt? tree bes : : oe sided pens hala ote ot eels 1 te om Pokey das? G) earn ; ‘ eel fa ‘Matas Pantie wmEsINNy - 1 ae aniell _Egete aor eae tl See tte ioe ci * oe ef mks iqeby geile’ pits oo vaew i Ab AN cages * . x a ; = “lh Apa Ste) bi ude ' vas ia ee i ed ee Aer em ; 7 x =. faite s _ a ' an ——- es 1 — i ba (tei =m ¢ >A fee id woe = « a e~ 1% the > g as AK or (Reports 87 through 89 are in one cover.) 87. Ethyl-p-aminobenzoate (Benzocaine): Efficacy as an Anesthetic for Five Species of Freshwater Fish, by V. K. Dawson and P. A. Gilderhus. 1979. 5 pp. 88. Influences of Selected Environmental Factors on the Activity of a Prospective Fish Toxicant, 2-(Digeranyl-amino)-ethanol, in Laboratory Tests, by C. A. Launer and T. D. Bills. 1979. 4 pp. 89. Toxicities of the Lampricides 3-Trifluoromethyl-4-nitrophenol (TFM) and the 2-Aminoethanol Salt of 2’,5-Dichloro-4’- nitrosalicylanilide (Bayer 73) to Four Bird Species, by R. H. Hudson. 1979. 5 pp. (Reports 90 and 91 are in one cover.) 90. Accumulation and Loss of 2’ ,5-Dichloro-4’-nitrosalicylanilide (Bayer 73) by Fish: Laboratory Studies, by V. K. Dawson, J. B. Sills, and Charles W. Luhning. 1982. 5 pp. 91. Effects of Synergized Rotenone on Nontarget Organisms in Ponds, by R. M. Burress. 1982. 7 pp. (Reports 92 through 94 are in one cover.) 92. Acute and Chronic Toxicity of Rotenone to Daphnia magna, by J. J. Rach, T. D. Bills, and L. L. Marking. 1988. 5 pp. 93. Toxicity of Rotenone to Developing Rainbow Trout, by T. D. Bills, J. J. Rach, and L. L. Marking. 1988. 3 pp. 94. Oral Toxicity of Rotenone to Mammals, by L. L. Marking. 1988. 5 pp. NOTE: Use of trade names does not imply U.S. Government endorsement of commercial products. protecting our fish and wildlife, preserving th of our national parks and historical ple life through outdoor recreation. 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