? 4... - ”I? I! f 3‘” i am; .ézgmzzzmzmw. 31W" fl}, gamma 395% WWW v 11.34....‘0 .1...” ..-w - .q . «.4 «q; "a.“ 5..----‘._ . 1 ~(,'l.'/(.". . .‘ a -;J.-‘..-.-..-.. , ,-,.‘.- ._ Aflu04 summ Hmumg T manuscm puma Hmuuflum m capo: m: :K\ xmmamlwcflxwz 1;. \\ mowowuowmcH Hmumz Pyrex columns, 2.4 x 50 cm, fitted with a fritted-glass disk, were packed with 10 g of Florisil-Celite (5:1) with a layer of anhydrous Na2804 above and below the packing. The Florisil, activated at 649°C by Floridin, Inc., was deacti- vated with approximately 10% distilled water and the mixture was calibrated before use to ensure conformation to the elu- tion procedure used. Each sample was eluted with 200 ml of .grhexane and then reconcentrated to a volume of 5 ml. These extraction and clean-up procedures generally follow those recommended by Shell Development Company (1964) with several modifications. One liter water samples, taken periodically from the out- lets of the test aquaria, were extracted successively in 2 l separatory funnels with 100, 50, 50, 50, and 50 ml of hexane. The combined extract was dried over anhydrous Na2804 and concentrated to 5 ml for introduction into the gas chromatograph. All solvents were re-distilled before use. A Beckman-GC 4 chromatograph. equipped with a discharge electron capture detector was used for the analyses. It was fitted with a 6 ft (1.85 m) x 1/16 in (1.59 mm) Pyrex column packed with 11% QF-1 and 5% DC 200 on Gas Chrome 0 (60/80 mesh) and was operated at a column temperature of 220°C and a 50 ml/min helium (99.995% pure) flow. The injection temperature was 250°C and the detector temperature, 275°C. Standards were injected at the beginning of each run, after every six samples, and at the end of the run. Quantitations were based on peak height and the concentrations were based on the wet weight of the mussel. The identities of the insecticides and their metabolites found were confirmed gas chromatographically using columns packed with 5%:DC 11 on Gas Chrome Q and 11% QF-1-OV-17 (1.5:1) on Gas Chrome 0. Selected samples were also spotted on Brinkman pre—made silica gel thin layer plates, developed with hexane-diethyl ether (4:1) and detected with Rhodamine B. The insecticides used were DDT (2,2-bis(p-chlorophenyl)- 1,1,1-trichloroethane), obtained from City Chemical Corp., N. Y., N. Y., and dieldrin(hexachloroepoxyoctahydro—endo,exo- dimethanonaphthalene), obtained from Shell Chemical Co., N. Y., N. Y. Both were recrystallized and 99+% pure. Recovery for DDT from lake water was 89.214.4% and from tapwater was 81.015.8%. For dieldrin recovery from lake water was 69.112.4% and from tapwater was 69.0:2.5%. Each per cent recovery was based on four spiked samples. RESULTS AND DISCUSSION Mussels are held as biological monitors of pesticides during all seasons in streams with a broad range of water quality. To determine some of the effects these conditions had on uptake and elimination of pesticides three different series of experiments were run in which freshwater mussels were exposed to known concentrations of DDT or dieldrin. In the first series of tests two species of mussels were ex- 'posed to various concentrations of DDT in distilled water with 50 ppm Ca++ added as CaClg for shell maintenance. The purpose of this series of tests was to determine the up- take and elimination of the insecticide under conditions where there were no stimuli to feed. This condition could occur in small cold streams which have very little plankton or sus- pended material and do not naturally support mussel popula- tions. In the second series of tests, mussels were exposed to various concentrations of DDT or dieldrin in lake water to ascertain the uptake and loss of the insecticides under simulated natural conditions. The third series involved the exposure of the mussels to DDT or dieldrin at different temperatures to determine 1O 11 the effect of temperature on the up-take and loss of the insecticides. Reconstituted Distilled Water For the first experiment (1) thirty mussels (Elliptio dilatatus) were collected from the Looking Glass River. These were held for several months without food at 9°C before being placed in the test aquaria. Three control mussels were analyzed for pesticides immediately prior to the test and found to contain low levels of DDT and its metabolites TDE and DDE (Table 1). The remaining mussels were placed into the test aquaria and exposed to a mean concentration of 0.5810.11 ppb DDT for three weeks at 2011°C. After the intro- duction of DDT was stopped the mussels were exposed to recon- stituted distilled water for one additional week. DDT at a concentration of 0.14 ppb was still present in the water leaving the aquaria at the end of the fourth week. Consider- able mortality (50%) occurred during the experiment, and, for this reason, the experiment was terminated after the fourth week." Three mussels were removed from the aquaria after each week and analyzed for insecticide content. The total DDT and its metabolites, TDE and DDE increased nearly fourfold after one week's exposure, continued to increase during the second week and then remained about the same for the following week Table 1. 12 Concentrations.of DDT and its metabolites (ppm, wet weight)in Elliptio dilatatus exposed to 0.58:0.11 ppb DDT at 20°C for three weeks. Weeks Weight Fat DDE TDE DDT Total (9) (per cent) 0 17.890 0.70 0.022 0.010 0.009 0.041 18.848 1.10 0.029 0.024 0.048 0.101 10.481 1.14 0.018 0.011 0.017 0.045 Mean 0.98 0.025 0.015 0.022 0.062 1 17.785 0.78 0.014 0.115 0.105 0.251 11.579 1.08 0.018 0.105 0.102 0.228 11.908 1.25 0.010 0.100 0.148 0.255 Mean 1.02 0.014 0.108 0.118 0.257 2 19.922 1.04 0.009 0.187 0.228 0.421 17.508 1.07 0.008 0.105 0.121 0.252 12.180 1.17 0.009 0.218 0.548 0.571 Mean 1.10 0.008 0.189 0.251 0.408 5 14.917 0.80. 0.008 0.185 0.257 0.408 12.201 0.75 0.005 0.175 0.128 0.508 8.955 0.50 0.018 0.298 0.147 0.485 "Men 0.97 0.010 0.211 0.171 0.592 Stop DDT introduction 4 .12.586 1.00 0.008 0.189 0.154 0.551 14.854 1.15 0.007 0.244 0.079 0.529 10.982 0.70 0.009 0.228 0.155 0.588 Mean 0.95 0.008 0.220 0.128 0.558 13 (Table 1). The concentration in the mussels decreased about 10% after the introduction of DDT was halted (Table 1). The increase in total DDT and metabolites was due to an increase in DDT and TDE as the concentration of DDE decreased from that of the controls. There was no reduction in the per cent fat of the mussels even though they were being starved during the experiment. In the second experiment (2) two changes were made in order to solve the mortality problem. The temperature was decreased from 20° to 15°C and the flow rate was increased from 50 ml/min to 120 ml/min. For this experiment (2) 40 mussels (Anodonta grandis) were collected from the Looking Glass River just prior to experimentation. Three mussels were analyzed for pesticide content and were found to have 5 to 10 times the concentration of DDT found in any of the other mussels used in these experi- ments (Table 2). The remaining mussels were exposed to a mean concentra- ; tion of 0.2710.08 ppb DDT for a period of five weeks. The experiment was continued for four weeks after the introduc- tion of DDT was terminated. The DDT concentration in the water decreased to 0.09 ppb at the end of the sixth week and traces ( N 00.55 500.0 000.0 000.0 500.0 00.0 50mmsE 05053 000.0 000.0 m05s5m 55.0 050.0 050.0 500.0 000.0 50.0 15080505nnnz 00.05 000.0 000.0 000.0 500.0 00.0 05omsz 00.0 000.0 000.0 050.0 000.0 00.5 0500m5> 5 00.0 000.0 050.0 500.0 000.0 00.0 5mmmse 050:3 000.0 000.0 mm05550 05.0 050.0 000.0 000.0 000.0 55.0 i5vaa5eanenz 00.5 500.0 050.0 000.0 000.0 05.0 5050052 05.0 000.0 000.0 000.0 000.0 05.5 nmumom5> 0 50509 50508 900 was man 500 mmsmm5a mxmmz “50503 00.5 550503 503 5:00 .500 .mx003 5505 500 U 005 um 900 £00 00. 0550. 0 05 ommomxm 050cmmm macovocd :5 AEQQV mmu550Q05mfio m55 Ucm BOG mo mCO5umuucmocoo £002 .0 05908 19 .05ucme 0am 05550 000:5Uc55 .mc05umuu Ic0ocoo 500058 05033 :5 000550c5 5020 .050055 0>550500500H 0cm 0>5um0050 002555000 .00500 005350550 0005:: 0500058 005:5 mo c0020 00.05 000.0 005.0 505.0 000.0 00.0 500008 05003 000.0 000.0 . 005050 00.00 005.0 000.0 005.0 000.0 50.0 805050002 00.50 500.0 005.0 005.0 000.0 50.0 050052 05.00 500.0 005.0 000.0 000.0 55.5 000005> 0 00.00 505.0 550.0 005.0 550.0 00.0 500008 05003 000.0 000.0 005550 00.55 000.0 005.0 505.0 000.0 00.0 805000002 00.00 505.0 000.0 500.0 050.0 50.0 050002 50.00 000.0 005.0 000.0 000.0 00.5 000005> 0 00.50 055.0 500.0 555.0 050.0 00.0 500005 05003 000.0 000.0 005050 05.00 055.0 500.0 000.0 500.0 05.0 100205000002 05.05 500.0 550.0 505.0 000.0 00.0 0500:: 00.50 050.0 005.0 550.0 000.0 00.5 000005> 0 005005005505 and 0050 00.05 055.0 000.0 005.0 550.0 00.0 500008 05003 000.0 000.0 005050 50.55 000.0 555.0 500.0 000.0 05.0 500505000002 00.00 005.0 050.0 055.0 050.0 00.0 0500:: 00.05 500.0 000.0 005.0 000.0 05.5 000005> 5 50005 50509 and was man 500 0000055 05003 500503 500 500503 003 5:00 500 009:5ucooll0 05509 20 insecticide in the muscle portion to the viscera as the experiment progressed. This indicates some movement of the insecticide from the gills and mantle where it was initially absorbed to the viscera. The tissue fluids generally contained about ten times the concentration of DDT present in the water, considerably less than any of the tissues and neither TDE or DDE were detected. As in the two previous experiments the concentrations in the mussels dropped very slowly after the introduction of DDT was stopped. However, the ratio of DDT to TDE decreased indicating that, although the insecticide was not being eliminated, it was being converted to TDE (Table 5). The same trend, but not as pronounced, was also observed in the second experiment (Table 2). Again the fat levels remained about the same throughout the experiment (Table 5). In the final experiment (4) with distilled water the insecticide concentration was reduced to 0.08:0.02 ppb. The mussels (Anodonta grandis) which were collected from the Red Cedar River and held in the laboratory for several months at 9°C, were exposed to this concentration for four weeks at 15°C. Three control mussels were analyzed initially and three mussels were analyzed after each week. The results show very little increase in insecticide concentrations from those of the controls (Table 4) suggesting that DDT levels in the 21 Table 4. Concentrations of DDT and its metabolites (ppm, wet weight) in Anodonta grandis exposed to 0.0810.02 ppb DDT at 15°C for four weeks. Weeks Weight Fat DDE TDE DDT Tota l (g) (per cent) 42.801 0.51 0.006 0.009 0.016 0.051 52.285 0.51 0.007 0.009 0.017 0.055 9.159 0.60 0.017 0.025 0.058 0.080 Mean 0.54 0.010 0.014 0.024 0.048 59.070 0.55 0.008 0.014 0.027 0.049 49.056 0.59 0.005 0.014 0.015 0.054 19.699 0.51 0.011 0.027 0.015 0.054 Mean 0.47 0.008 0.018 0.019 0.046 69.507 0.56 0.004 0.010 0.009 0.024 54.789 0.68 0.007 0.021 0.057 0.065 18.247 0.48 0.008 0.022 0.019 0.050 Mean 0.51 0.007 0.018 0.022 0.046 50.557 0.55 0.008 0.017 0.057 0.062 41.590 0.61 0.007 0.018 0.015 0.040 18.949 0.79 0.010 0.025 0.052 0.068 Mean 0.64 0.008 0.020 0.028 0.056 28.656 0.75 0.007 0.019 0.024 0.049 28.860 0.64 0.009 0.054 0.025 0.069 21.576 0.55 0.011 0.040 0.029 0.080 Mean 0.65 0.009 0.051 0.026 0.066 22 Red Cedar were probably around 0.08 ppb in the area where the mussels were collected. The concentration factor (mussel: water) here was slightly less (9a, 700 vs 1000) than the factor for the previous experiments. The same general pattern, an initial, rapid uptake, subsequent leveling—off, and a very slight decrease when DDT introduction was terminated was seen in each of the first three experiments (Figure 2). The large difference between the equilibrium concentration of the second experiment (2) and the other two (1 and 5), deSpite the very similar mean water concentrations of DDT in the three experiments, is dif- ficult to explain. The main differences were in the initial "physiological state" of the mussels and the species of mussel. The mussels in the second experiment had much higher concentrations of DDT when collected (0.220 vs 0.062 and 0.027 ppm) and were placed in the test aquaria a few days after collection while the others were held for considerable lengths of time at 90C before experimentation. The initial concentra- tion should in theory have no effect on the equilibrium con- centration unless it is higher than the equilibrium concen- tration. In distilled water there seems to be a very slow elimination of DDT so a high initial concentration in the mussel could affect the equilibrium concentration. The fact that the mussels in the second experiment were fresh from natural conditions might result in greater filtering activity initially than those mussels conditioned to no food and cold 25 .umumz omaawpmflp pmuouwumcooou CH Ban Add mm.o ou hm.o ou Ummomxm mammmse :H mmpwaonmuwe mpg cam Baa mo mcofiumuucmocoo cmmz .N musmflm 24 Amy .ucmfiwummxm 703 b p mxmmz ~<fl ~10 '-C\J bx—l 3v unmEMHmmxw N musmam :3 unwedummxm No.0 .mo.o udo.o umo.o nmo.o lfioo ..~.o um.o 4.0 .m.o (qqfiram 39M) mdd 'qu Iago; 25 temperatures prior to experimentation. Thus they may have concentrated the insecticide to a greater extent because of greater contact. The difference in species did not correspond with the difference in equilibrium concentrations as Elliptio in experiment (1) and Anodonta in experiment (3) reached similar concentrations while Anodonta in (2) was much higher. The absence of differences between species of mussels in concen— trating pesticides was also found in previous studies (Bedford 33 al,, 1968). In all of the experiments run in distilled water profuse filamentous fungal colonies developed on the aquaria walls, substrate, and mussel shells. Presumably the fungi were living on the waste products of the mussels. In the last two experiments (5 and 4) the growths were especially abundant and samples were collected on Whatman No. 1 filter paper, dried, weighed and extracted with hexane. The fungi samples were found to contain ten times the concentrations of DDT and its metabolites found in the mussels (Tables 5 and 6). When the introduction of DDT was terminated in the third experiment (5) the DDT concentration in the fungi remained about the same for one week and then dropped sharply the following week (Table 5). In the last experiment (4) the levels were approximately the same for all three samples of fungi and were slightly lower than the final concentration in the third experiment (5). This follows, as the DDT 26 Table 5. Concentrations of DDT and its metabolites (ppm, dry weight) in fungal growth removed from test aquaria during Experiment 5. Weeks weight DDE TDE DDT Total ""' (g) 5 0.1704 1.995 8.509 124.119 154.625 4 0.4065 1.555 4.554 105.781 111.488 5 0.1665 2.706 8.720 120.260 151.686 6 0.1495 0.656 2.009 22.458 25.084 7 0.0590 0.952 4.746 16.949 22.627 Table 6. Concentrations of DDT and its metabolites (ppm, dry weight) in fungal growth removed from test aquaria during Experiment 4. Weeks Weight DDE TDE DDT Total (9) 2 0.1497 1.169 2.458 14.696 18.505 5 0.1649 0.940 0.910 15.645 15.494 4 0.1749 0.715 0.457 15.057 16.209 27 concentration in the water was still slightly higher (0.10 ppb) at the end of the third experiment (5) than the mean concentration (0.08 ppb) of the last experiment (4). The presence of the fungal growth offers a possible explanation of the relatively high levels of DDT remaining in the water for three weeks following the termination of DDT introduction. The fungi apparently released considerable DDT when the introduction was stopped unless they metabolized it to a non-detectable metabolite. The release of DDT by the fungi could also have occurred in the second experiment (2) and may explain the increase in DDT concentrations in the mussels the week following termination of the DDT intro- duction. Lake Water Lake Lansing, a shallow, warm water, eutrophic lake located in Ingham County, Michigan, provided the water for this series of experiments. Analysis showed that the water contained small amount of DDT (<10.05 ppb) but no detectable dieldrin. Water was pumped periodically to a stainless steel tank which fed a constant-head tank. This provided a con- stant flow of lake water to the mixing flask where the insecticide solution was metered. Mussels (Lampsilis siliquoidea) from Gun Lake were used in the first two experiments and were collected just prior 28 to the tests. In the first experiment (5) the mussels were exposed to a mean concentration of 0.57i0.12 ppb dieldrin for three weeks and then the concentration was approximately doubled to 1.16:0.15 ppb for one week. Introduction of dieldrin was then terminated and the mussels exposed to Lake water for three additional weeks. One week following the termination of the dieldrin introduction the insecticide level in the water had decreased to 0.11 ppb in the tank overflow and at the end of the experiment it was 0.05 ppb. Three control mussels were analyzed at the beginning of the test and found to have small concentrations of dieldrin (Table 7). Three mussels were removed from the test aquaria each week and analyzed for insecticide content. The mussels appeared to reach equilibrium concentrations in one week or less as the highest concentration was attained after one week and then decreased slightly the following two weeks (Table 7). When the water concentration of dieldrin was doubled the concentration in the mussels also doubled (Table 7). When the introduction of dieldrin was stopped the di- eldrin concentrations in the mussels dropped very rapidly for the first two weeks and then remained nearly constant (Table 7). This was in contrast to the experiments with DDT in distilled water when after introduction of DDT was termi- nated the concentrations in the mussels remained approximately the same. 29 Table 7. Concentrations of dieldrin (ppm, wet weight) in Lampsilis siliquoidea exposed to 0.57iO.12 ppb dieldrin in lake water at 20°C for three weeks. Weeks Weight Fat Dieldrin (g) (per cent) 0 16.501 0.71 0.018 7.566 1.08 0.011 5.162 1.25 0.000 Mean 1.01 0.010 1 9.599 1.18 0.677 6.782 1.01 0.644 5.111 1.06 0.775 Mean 1.08 0.699 2 12.701 0.62 0.456 4.657 1.06 0.754 5.429 1.00 0.828 Mean 0.89 0.672 5 8.714 0.99 0.545 5.418 1.15 0.705 4.504 1.10 0.587 Mean 1.08 0.612 4 5 Mean Mean 12.715 6.667 5.175 Stop dieldrin introduction 12.217 7.645 5.650 0.75 0.91 1.08 0.91 0.65 0.90 0.96 0.85 Water concentration increased to 1.16iO.15 ppb dieldrin. 1.115 1.485 .1.467 1.555 0.504 0.560 0.258 0.501 continued 50 Table 7-—continued Weeks Weight Fat Dieldrin (g) (per cent) 6 14.087 1.01 0.052 5.582 0.80 0.082 1.952 1.57 0.128 Mean 1.06 0.088 7 2.678 2.29 0.080 4.566 1.50 0.084 2.525 1.02 0.056 Mean 1.54 0.075 51 A semi-log plot of the dieldrin residue values following stoppage of the dieldrin introduction, were linear with a negative slope (Figure 5). The equation for this relationship is as follows: (1) y = (A) (-BX) where: Y = concentration of dieldrin in the mussel (ppm, wet weight) A = regression intercept (ppm) X = weeks following cessation of dieldrin introduction B = relative rate of residue loss or for the rectified data: (2) Log Y = log A - (log B) x Log B, the regression coefficient, estimates the log- arithmic rate of residue loss. In this case the log of the dieldrin concentration in the mussel decreased at an esti- mated uniform rate of 0.457 and the residue half-life calcu- lated from equation (2) was 4.7 days. If one eliminates from the regression analysis the third week values, where the mussels appear to be in equilibrium with the dieldrin that still remained in the water, the log of the concentration in the mussels decreased at an estimated rate of 0.607. The half—life value is then decreased to 5.5 days. In the second experiment (6) the mussels (Lampsilis siliquoidea) were exposed to a mean dieldrin concentration 52 Figure 5. Loss of dieldrin from Lampsilis siliquoidea in lake water. Dieldrin, ppm (wet weight) 55 '/ Tfi = 5.5 days 0.04- Log B = 0.607 0.051 0.02. 0.01 . . <5 i 2 5 Weeks Figure 5 54 of 0.0610.01 ppb for three weeks. The control mussels con- tained no detectable dieldrin. As with the first experiment (5) the mussels attained their highest dieldrin concentration after the first week's exposure (Table 8). The concentration in the mussel then dropped about 40% the following week and rose to a level midway between the first and second after the third week. In both experiments the fat levels remained relatively constant. Equilibrium concentrations in the mussels were plotted against the corresponding water concentrations and fitted with a least squares line (Figure 4). The slope of the line or regression coefficient was 1.19 with the concentration in the mussel in ppm and the water concentration in ppb. Thus the mussels were concentrating the dieldrin about 1200 times over the water concentration. The correlation coefficient (r) of 0.976 indicated a very close fit and a highly significant correlation between the concentrations (Fe = 576.8, F = 995 10.1). However the assumptions of variance homogeneity and independence of means and variances were not met due primarily to the small mean and variance of those mussels exposed to 0.07 ppb dieldrin. The same experimental apparatus used for the dieldrin experiments was employed for DDT in lake water. Mussels (Anodonta grandis) were collected from the Red Cedar River and exposed to three different concentrations of DDT. 55 Table 8. Concentrations of dieldrin (ppm, wet weight) in Lampsilis siliquoidea exposed to 0.06i0.01 ppb dieldrin in lake water at 20°C for three weeks. Weeks Weight Fat Dieldrin (g) (per cent) 0 12.185 0.65 0.000 5.450 0.89 0.000 4.210 1.07 0.000 Mean 0.86 0.000 1 15.719 0.65 0.055 4.776 1.01 0.071 5.846 0.84 0.081 Mean 0.85 0.068 2 6.504 1.55 0.055 4.915 1.15 0.050 5.779 0.96 0.056 Mean 1.15 0.040 5 9.868 0.89 0.041 5.750 1.15 0.059 5.105 0.84 0.060 Mean 0.96 0.055 56 .©o>oEoH mum? won“ QUflSS Eoum uwumz on» GM mcowumuucmocoo on» cam Ammoflosqflawm mflHMmQquv mammmsfi CH mcowumuucmocoo cwuoamflo cooSDmQ coaumamm .¢ ouomwm 57 0.0 ,flmm:.nwum3 Ca cfluwamwa 0.0 5.0 who a ousmwm 0.0 :6 I N o O r N) O l d‘ O I LO Q ' LO 0 O I [x o O 10.0 I O) C) (qutem gem) mdd 'sIassnm u: urxptera lo. .fi 1 I N H o o x—{ H r N) r! F fi' x—l LI 0 H 58 In the first experiment (7) the mussels were exposed to a mean concentration of 0.6210.15 ppb DDT for three weeks followed by five weeks exposure to lake water after the cessation of DDT introduction. The DDT concentration in the water dropped to 0.24 ppb at the end of one week and was down to 0.11 ppb three weeks after termination of the introduction. The mussels were dissected before insecticide analysis as in the third distilled water experiment (5) except that the body fluids were not analyzed. The control mussels were found to have very low DDT residues (Table 9). After one week's exposure the concentration in the mussels increased sharply to over 1 ppm and stayed at that level the following week before increasing again after three weeks's exposure (Table 9). The sudden increase in DDT residue concentrations after three weeks was due to excessive concentration of DDT in the test tanks during a 2 1/2 day period when the dilution water line became plugged with debris and periphyton. Unfortunately no water samples were obtained when this problem was discovered. Later in the week, following clearing of the line, water samples showed concentrations of DDT to be only slightly higher than the mean for the three weeks. The concentrations of DDT and metabolites were again much higher in the viscera than in the muscle, mantle, and gills (Table 9). However, in contrast to the distilled water experiment, the marsupia had very high concentrations of insecticides, in most cases nearly as high as the viscera. 59 Uoscflucoo mo.w¢m mm>.fi mmm.a mma.0 mm0.0 m>.0 Hummus oaosz H¢.H0m mmm.d mmw.d >mm.0 mm0.0 $0.0 ANVEswmomumS m¢.m¢H «00.0 >¢m.0 moa.0 mm0.0 50.0 odomsz >¢.Nmm d¢>.m mmm.m mmm.0 000.0 mm.0 mqumH> a coquDUouucfi BOD moum wm.0mm m>¢.N mmm.m mmd.0 mw0.0 mm.0 Hmmmse waonz o~.omm cma.m mam.m mam.o mmo.o 55.0 invasumsmumz mm.m¢m wm>.a >mm.a N0d.0 mm0.0 05.0 odomoz m>.mmm emm.m m>0.m wam.0 mm0.0 no.6 mumomw> m mm.mmd moa.a Hem.0 wad.0 >¢0.0 0m.0 Hmmmos maoaz Hm.mmm mme.d mm¢.a mmd.0 N00.0 >>.0 AHonHmsmnmz ¢N.Nm H0>.0 mmm.0 «no.0 mmo.0 m>.0 odomsz mm.wm« mwe.d dmm.d m¢H.0 Nm0.0 $0.6 mnmomw> N em.oma mmm.d mnm.d Hm0.0 mm0.0 mm.0 Hmmmoa macs: mm.mmd «60.0 mmm.0 mn0.0 md0.0 ¢¢.0 AHVEsHmsmHmz mm.m¢d «No.6 mmm.0 H50.0 0m0.0 05.0 manna: am.mdd mmm.a mvm.d mad.0 mm0.0 mN.H mumomw> a «0.0 ¢N0.0 0d0.0 $00.0 0d0.0 no.0 Hmmmse maoa3 md.m >a0.0 >00.0 moo.0 000.0 am.0 vaomsz md.¢ «no.0 mdo.0 m00.0 ma0.0 m>.0 mmnmumw> 0 Hmpoa Hmuoa son was moo “mm mmsmmue mxmmz assume umm usmums umz ucmo “mm .mxmm3 mounu How ooom um Hmum3 mme CH 900 Ram ma.0HNm.o ou vwmomxo mflocmum mucooocd cw Aemmv mmuaaonmuwfi mum 0cm BOG mo mcowumnucmocoo cmmz .m magma 40 .maucmfi 0cm 04440 mmflsaosH¢ .osmmflu o>4ummmwv 0cm m>4uoocoummu Umswnfioon 0U0#OC 0mH3H®£MO mmmHCD mHmmmDE mmhflu. MO Gmwzm 00.04 004.0 040.0 000.0 000.0 00.4 400000 04003 05.00 000.0 040.0 050.0 400.0 00.0 040002 00.40 400.0 004.0 004.0 000.0 00.4 000004> 0 00.55 045.0 400.0 044.0 000.0 00.0 400000 04003 04.00 000.0 055.0 000.0 440.0 50.4 440004000002 50.00 400.0 044.0 400.0 400.0 40.0 040002 04.50 040.4 000.0 004.0 000.0 00.4 000004> 5 44.004 000.4 040.0 044.0 000.0 55.0 400000 04003 00.000 000.4 004.4 000.0 400.0 04.0 400004000002 40.00 005.0 400.0 450.0 000.0 55.0 04000: 00.054 405.4 454.4 404.0 000.0 00.0 000004> 0 05.004 000.4 404.4 504.0 440.0 00.0 400000 04003 44.004 000.0 540.0 044.0 500.0 55.0 04000: 00.000 000.0 445.4 000.0 400.0 50.0 000004> 0 40009 40009 9cm mo9 man 000 000049 04003 0.00403 00.4 ”£0403 003 0000 40.4 Umsc4ucoollm magma 41 When the concentration of insecticide in the mussels was placed on a fat weight basis the difference in concentration in the muscle and viscera portions was much reduced (Table 9). The concentrations in the marsupia, when placed on a fat weight basis, were in some cases considerably higher than the DDT concentration in the viscera owing to the relatively low fat content of the marsupia. When the introduction of DDT was terminated the mussels showed a steady decline in insecticide concentrations (Table 9). When plotted against time on semi-log paper the levels in the mussels decreased in a linear fashion as occurred with dieldrin (Figure 5). When equations (1) and (2) were applied, the regression coefficient (log B) for total DDT and metabo- lites was found to be 0.148 and the half-life was 15.6 days. Thus the half-life of DDT was between three and four times longer than the half-life of dieldrin in the mussels. Gakstatter and Weiss (1967) also found a much slower elimina— tion of DDT than dieldrin and lindane in fish. They concluded that uptake and elimination rates were related to the solu- bility of the insecticide in water. Grzenda gt_al. (1970) reported a logarithmic rate of loss of 0.0725 and a half-life of 29.5 days for DDT in goldfish. This is about half the elimination rate in mussels. However, the initial body burden was produced in the fish by feeding them contaminated food rather than via the water. This could account for some of the difference, at least initially for most of the fed 42 Figure 5. Loss of DDT from Anodonta grandis in lake water. 45 L5 14 .05 5 Ac V. a d no 64. .1. 5 o 10 __ = 1 mm B 9 O L r). .p 7 , _ 4 4 4 4 “ fl. ,.- no 4 J 14 u .. q q l. nw nw 4N nu 00.8 7. no .5 .4 .5 ?. 01 0 0 O ,0 O G O O O 4 5 2 100 0 0 O 0 0 0 0. 4950493 umBV Ema . BOO HMNOB Weeks Figure 5 44 insecticide was probably incorporated into the fish tissue while it was possible a significant proportion of the residue level in the mussels was simply adsorbed onto the gills and mantle and was thus more easily lost. The greater fat con- tent (5-29%) and initial body burden (1.2-18.4 ppm) in the fish could also have an effect on the elimination rates. In the next two experiments (8 and 9) mussels (Anodonta grandis) were exposed to two different concentrations of DDT for two week periods. In the first test (8) the mussels were exposed to a mean concentration of 0.42:0.12 ppb DDT. The residue levels increased greatly after one week and then leveled off in the mussels (Table 10). In the second test (9) a mean concentration of 0.14:0.12 ppb DDT was maintained in the water. The concentration again increased from the controls after one week and then did not change the following week (Table 11). The equilibrium concentrations for DDT in the mussels were plotted in a manner similar to the dieldrin results and fitted with a least squares line (Figure 6). The regression coefficient or slope of the line was 2.56 with the concentra- tion of DDT in the mussels in ppm and the water concentration in ppb. The mussels were thus concentrating the DDT about 2400 times the concentration in the water or about twice the degree of concentration for dieldrin. The correlation coefficient (5) of 0.880 was not quite as high as that for dieldrin but still indicated a close fit. 45 Table 10. Concentrations of DDT and its metabolites (ppm, wet weight) in Anodonta grandis exposed to 0.42:0.12 ppb DDT in lake water at 20°C for two weeks. Weeks Weight Fat DDE TDE DDT Tota l (g) (per cent) 0 70.642 0.45 0.005 0.005 0.025 0.055 65.887 0.66 0.005 0.004 0.020 0.029 57.712 0.82 0.005 0.007 0.020 0.050 Mean 0.64 0.004 0.005 0.022 0.051 1 89.721 0.54 0.015 0.074 0.555 0.624 75.109 0.44 0.014 0.051 0.417 0.482 52.155 0.74 0.022 0.105 0.972 1.099 Mean 0.50 0.017 0.077 0.641 0.755 2 105.555 0.40 0.021 0.095 0.726 0.859 64.754 0.69 0.051 0.122 0.858 0.990 75.414 0.48 0.017 0.085 0.617 0.178 Mean 0.52 0.025 0.100 0.727 0.850 46 Table 11. Concentrations of DDT and its metabolites (ppm, wet weight) in Anodonta grandis exposed to 0.14i0.02 ppb DDT in lake water at 20°C for two weeks. Weeks Weight Fat DDE TDE DDT Total (9) (per cent) 0 44.815 0.48 0.002 0.006 0.055 0.042 1 67.275 0.64 0.050 0.055 0.078 0.145 52.750 0.55 0.006 0.016 0.066 0.088 46.970 0.68 0.006 0.015 0.080 0.102 Mean 0.62 0.014 0.022 0.075 0.111 2 87.602 0.51 0.007 0.014 0.066 0.087 67.497 0.48 0.011 0.022 0.100 0.155 55.720 0.58 0.007 0.014 0.065 0.084 Mean 0.46 0.008 0.017 0.076 0.101 Figure 6. 47 Relation between DDT concentrations in mussels (Anodonta grandis) and the concentrations in the water from which they were removed. 48 1.74 d d d 4. 5 2 1 1 .1 0 98 7 6 5 1 00 0 0 0 1.1‘ Apnmwwz 903v Ema .mammmoe :4 800 40008 0.5‘ 0.2- 0.1- 0.0« -0.1- 071 0.4 0.5 0.6 DDT in water, 0.5 0.2 -0.2 ppb 49 The correlation between the concentrations was again found to be highly significant (F = 54.76, F = 10.58) exp. .995 but, as was the case with dieldrin, the assumptions of vari— ance homogeneity and independence of the means and variances were not met owing to the relatively low variance of the mussel concentrations at the 0.14 ppb water concentration. During both the DDT and dieldrin experiments in lake water several water samples were filtered through Whatman No. 1 filter paper and both the filtrate and the residue were analyzed in order to determine the relative distribution of the insecticide that had been metered into the lake water. The majority of the dieldrin was found in the suspended matter while the DDT was fairly equally distributed between the water and suspended matter (Table 12). This result is somewhat incongruous with the water solubilities of the two compounds. Robeck gt al. (1965) reported dieldrin to be several times more soluble than DDT yet a larger proportiOn of DDT than dieldrin was found in the water filtrate. Effect of Temperature Three experiments were run to determine the effects of temperature on the uptake and loss of DDT and dieldrin. Dechlorinated tap water was used as the water source and the insecticide was metered into a common mixing flask. The treated water was divided into the four aquaria maintained 50 Table 12. Distribution of DDT and dieldrin in lake water (ug/l of water). Suspended Water Insecticide Replicate matter filtrate Total Dieldrin 0.28 0.21 0.49 0.26 0.11 0.57 Mean 0.27 0.16 0.45 Unfiltered 0.42 DDT 0.26 0.56 0.62 0.52 0.52 0.64 Mean 0.29 0.54 0.65 Unfiltered 0.65 0.25 0.28 0.55 0.54 0.50 0.64 Mean 0.50 0.29 0.59 Unfiltered 0.64 51 at different temperatures via glass Y tubes and Teflon tubing. A flow rate of 550-400 ml/min was maintained through each aquaria. For the first experiment (10) 22 mussels (Anodonta grandis) were collected from the Red Cedar River, divided into groups of five, and acclimated to 50, 10°, 15°, and 20°C over a period of one week. Two controls analyzed for insecticide content were found to have low levels of DDT and its metabolites (Table 15). The remaining mussels were exposed to virtually the same concentration of DDT (0.55 to 0.65 ppb) for three weeks. Two mussels were removed from each aquaria after one and three weeks exposure, dissected into visceral and muscle- gill fractions as in previous experiments (5 and 7), and analyzed for insecticide content. The concentrations in the mussels in the 50C aquaria remained the same as the controls after one and three weeks exposure (Table 15). The mussels in the 100C and 150C aquaria both showed increases from the controls after one week. The mussels at 100C continued to increase after three weeks but those at 15°C seemed to level off (Table 15). The mussels in the 200C aquaria increased the greatest amount after one week but mortality of the remaining mussels at about two weeks eliminated the three week sample (Table 15). A two-way analysis of variance (from Li, 1964) was run to determine if the observed differences in total DDT levels 52 005544500 040.0 040.0 m00.0 400.0 04.0 400058 04053 040.0 m40.0 m00.0 400.0 00.0 040052 4N0.0 m40.0 400.0 N00.0 00.0 040004> mo.ofiwm.0 N.Qflm.4 m 400.0 480.0 N40.0 m00.0 45.0 400058 04053 004.0 800.0 440.0 400.0 00.0 040052 .I 040.0 mmo.0 m40.0 400.0 00.0 040004> mo.o+mm.0 0.040.04 000.0 400.0 000.0 000.0 mm.0 400058 04053 000.0 050.0 000.0 000.0 54.0 . 040002 000.0 mm0.0 «40.0 m00.0 00.0 040004> 50.0H0m.0 N.0H4.m4 000.0 mm0.0 000.0 400.0 00.0 400058 04053 mm0.0 440.0 800.0 N00.0 04.0 040052 m40.0 0m0.0 000.0 000.0 40.0 040004> 00.0H0m.0 4.000.04 NN0.0 040.0 000.0 000.0 mm.0 400058 04053 040.0 440.0 m00.0 400.0 04.0 040052 0m0.0 4N0.0 m00.0 400.0 48.0 040004> 00.0Hmm.0 oom.Qflm.4 4 040.0 040.0 n00.0 000.0 mm.0 400058 04053 440.0 040.0 m00.0 m00.0 00.0 4040052 000.0 040.0 000.0 000.0 45.0 0040004> 0 40408 800 mo8 man 405 0050048 5045044500500 0454 04003 4500 405 40003 10400808 .40403004 0040544045000 54 004540400804 450404440 40 Anmmv 8QD 04 00000X0 040504m mu50005¢ 54 4450403 403 .8000 00444090408 044 050 805 40 05040044500500 .04 04908 55 .040508 050 04440 000540544 .050040 0>4005004m04 050 0>4000m40 005448000 omHmmmDE 03¢ MO CMGZN 000.0 000.0 0N0.0 000.0 00.0 400058 04053 m>0.0 N40.0 0N0.0 000.0 00.0 040052 400.0 5N0.0 0N0.0 000.0 00.0 040004> 80.0400.0 N.0H4.04 000.0 000.0 040.0 400.0 04.0 400058 04053 004.0 050.0 4N0.0 000.0 04.0 040052 000.0 000.0 040.0 000.0 00.0 040004> 00.0400.0 4.0H0.04 0 40008 8CD 008 man 00m 050048 5040040500500 0450 04003 0500 405 40003 I0405808 005540500||04 04408 54 at different times (1 and 5 weeks) and temperatures (50, o, 15°C) were significant. The results showed no signifi— 10 cant difference between either temperature or time (Table 14). The concentrations attained at 15°C were much lower than one would expect judging from the earlier experiments in distilled water at 15°C. In contrast to the previous experiments, the concentrations in the muscle mantle and gills were as high or higher than in the viscera. Thus it appears that there was much less DDT incorporation into the tissue of the mussel. The mussels were observed to filter very little during the experiment (10). Perhaps this was due to their "physiological state" in the environment from which they were removed and/or insufficient acclimation time. The mussels were collected in midwinter at a water temperature of about 1°C. Another possible reason for this lack of filtering was the discovery of copper (0.05 ppm) in the water after the next experiment (11). Arthur and Leonard (1970) found that lower concentrations than this represented TLm values for the snail physa integra (0,059 ppm) and the amphipod Gammarus pseudo- limnaeus (0.020 ppm). Mussels (Lampsilis siliquoidea) for the following two experiments were collected from Gun Lake just prior to each experiment. The mussels were again subdivided and acclimated O to 5°, 10 , 15° , and 20°C. In the first experiment the mussels were exposed to mean concentrations of dieldrin from 0.97 to 1.12 ppb (see Table 15) for three weeks and the 55 Tabld 14. Results of an analysis of variance for the observed differences in mean concentrations of DDT and metabolites in Anodonta grandis with respect to length of exposure and temperature. Source SS DF MS Fexp. F.95 Temperature 0.00515 2 0.00157 6.58 19.0 Time 0.00009 1 0.00009 0.56 18.5 Error 0.00049 2 0.00025 Total 0.00571 5 lllll‘ 5.5.11! 5.5.15. I‘“.‘lll.ll>‘ ‘l/I.( [[{IIII 000.0 00.0 008.0 00.0 000.0 08.0 5002 000.0 00.0 008.0 00.0 800.0 80.0 000.0 00.0 080.0 08.0 080.0 08.0 080.0 00.0 088.0 00.0 800.0 88.0 0 000.0 00.0 088.0 00.0 080.0 00.0 000.0 80.0 5002 000.0 00.8 088.0 00.0 000.0 00.0 880.0 08.0 000.0 00.0 000.0 80.0 800.0 00.0 000.0 00.0 000.0 08.0 800.0 08.0 000.0 80.0 0 000.0 00.0 008.0 00.0 000.0 00.0 080.0 08.0 5002 000.0 00.0 008.0 00.0 000.0 00.0 080.0 80.0 000.0 00.8 008.0 00.0 000.0 00.0 000.0 00.0 6 000.0 00.0 088.0 08.8 000.0 08.0 000.0 80.0 8 5 000.0 00.0 5002 008.0 00.0 000.0 00.0 000.0 80.0 0 00000005 000 00000005 000 50000005 000 00000005 000 00003 0500 00m 0500 00m 0500 00m 0500 00m 08.0008.8 08.0H00.8 00.Qfl8o.8 00.0080.0 .0500 000030 0.000100 02000.00 0.9.100 0.00.0.0. 0 0030000500. 0 .000500000800 050000040 00 00003000 0000540045000 54 40000 054004040 00 00000x0 00040504440 044405800 54 5050403 003 .8000 54004040 00 5040000500500 .08 04508 57 .5008 0:0 00 00000000 0020 008.0 080.0 008.0 088.0 $08.0 008.0 000.0 008.0 800.0 000.0 000.0 000.0 00.0 00.0 00.0 00.0 80.0 00.0 00.0 08.0 80.0 00.0 80.0 00.8 800.0 080.0 000.0 000.0 008.0 008.0 008.0 0000.8 080.0 000.0 080.0 008.0 00.8 00.8 00.8 00.0 80.0 00.0 00.0 00.0 00.0 00.0 88.0 00.0 000.0 080.0 000.0 800.0 000.0 800.0 880.0 000.0 000.0 000.0 000.0 000.0 00.0 80.0 00.8 00.8 08.0 08.0 80.0 08.0 00.0 00.0 08.0 88.0 5002 5002 5002 fl 000000000000 00000000 0000 58 experiment was continued for another three weeks following termination of the dieldrin introduction. The level of dieldrin in the aquaria dropped to subdetectable levels (0.02 ppb) one week after termination. Three mussels were analyzed just prior to experimenta- tion for dieldrin content and one was found to have a rela- tively high dieldrin burden, about seven times the amount in the other two controls, which were quite low as had been the case for mussels previously collected from Gun Lake (Table 15). As was the case with DDT, the concentrations in the mussels at 50C remained about the same as the controls throughout the experiment. Both the 100 and 15°C mussels showed increases in insecticide content during the first three weeks followed by a slow decline after termination of dieldrin introduction with the concentrations in the 150C mussels about twice as high as those in the 100C mussels (Table 15). Problems of survival were again encountered at 200C, with the majority of the mussels dying in less than two weeks so that only one and two week samples could be taken. A few mussels were also lost at 150C (10%) but no mortality occurred in the So and 100C aquaria. The mussels that did survive at 20°C reached concentrations 2-5 times greater than those found in mussels at 15°C for the same length of exposure (Table 15). 59 A two—way analysis of variance to analyze for the sig— nificance of observed differences in insecticide concentra— tions with respect to time and temperature was again limited 0' and 15°C because of lack of survival of the 200C to 5°, 10 mussels for the three weeks. No significant difference was found for either temperature or time of exposure (Table 16). Following termination of dieldrin introduction at 15°C the residue data when plotted against time on semi-log paper were linear with a negative slope (Figure 7). Upon applica- tion of equations (1) and (2) the log of the dieldrin concen- tration in the mussels was found to decrease at an estimated uniform rate of 0.108 and the half-life was calculated to be 19.3 days. This rate of loss was much slower than found for the mussels in lake water at 200C (0.108 vs 0.457). Much of this difference is probably due to the lower temperature, however, as will be seen in the next experiment (12), the dieldrin concentration appeared to decrease slowly at 200C in tap water also. One sample (100C-5 weeks), which contained ten times the concentration of dieldrin of the other two replicates, was not included in computing the mean for that temperature and time because of suspected contamination. For the final experiment (12) two changes were made to enhance the survival of the mussels at 20°C. First the copper pipes in the laboratory building were bypassed with flexible plastic pipe. This reduced the copper concentration to 0.004 60 Table 16. Results of an analysis of variance for the observed differences in mean concentrations of dieldrin in Lampsilis siliquoidea with respect to length of exposure and temperature. Source SS DF MS Fexp. F.95 Temperature 0.05999 2 0.01999 5.58 6.94 Time 0.01852 2 0.00916 2.46 6.94 Error 0.01487 4 0.00572 Total 0.07519 8 61 Figure 7. Loss of dieldrin from Lampsilis siliquoidea in dechlorinated tapwater. Dieldrin, ppm (wet weight) 62 T% = 19.5 days Log B = 0.108 001 I 1 I 0 1 2 5 Weeks 65 ppm from 0.05 ppm. Second, the dieldrin concentration was cut in half. Three control mussels were analyzed and all contained low concentrations of dieldrin (Table 17). The remaining mussels were divided into the four aquaria after acclimation and exposed to mean concentrations of 0.42 to 0.45 ppb dieldrin for three weeks. The experiment was originally intended to study only uptake of the insecticide; however, since there was 100% survival of the mussels at all tempera- tures, enough mussels remained for another analysis. This sample was taken two weeks after termination of the dieldrin introduction at which time no detectable dieldrin remained in the water. After one week's exposure the mussels in the 50, 100, and 15°C all contained similar low levels of dieldrin only slightly greater than the controls (Table 17). Those at 200C contained about four times the amount of dieldrin found in the mussels at other temperatures (Table 17). At the end of two weeks there was little change from the one week concen- trations except at 50C where the level almost doubled. The mussels at 5°C again differed from those at the other tempera— tures after three weeks, only this time there was no change at 5°C and there was substantial increases (40 to 100%) at the other temperatures (Table 17). Two weeks following termination of the dieldrin intro- duction the mussels at all temperatures had lower concentrations 64 wmm.o mm.o wmo.o do.d dmo.o Om.o ano.o mm.o smwz mam.o mm.o dmo.o NH.H Had.o mm.o Hmo.o mm.o mmm.o Na.d mmo.o mo.d m>0.0 hm.o mmo.o mm.o «ma.o mm.o mmo.o >m.o «mo.o mm.o mmo.o om.o coHuUDUOHucH GAHUHmHU moum N>N.o hm.o omd.o mm.o Hoa.o mm.o mmo.o ¢m.o com: mmm.o mo.d hmm.o mm.o #mo.o mm.o moa.o om.o mom.o mm.o hoa.o mm.o moa.o mo.fi mmo.o No.d omN.o mm.o mad.o do.d mOfi.o ho.d m>0.0 Hm.o oom.o do.d amo.o mo.a «mo.o mo.d mmo.o mm.o cams mad.o oo.fi oma.o mm.d Nwo.o Nd.d wmd.o 00.0 mmH.o mo.d moo.o wa.a m¢o.o no.6 mmo.o mm.o mom.o do.d mmo.o wm.o hmo.o do.d mmo.o mm.o mmm.o m>.o Nmo.o v>.o m¢0.0 m>.o wmo.o m>.o com: mmm.o mm.o m»o.o m>.o mmo.o >m.0 who.o no.0 Nom.o mm.o dwo.o O>.o mmo.o 00.0 mmo.o mm.o omN.o mm.o m¢o.o m>.o omo.o om.o mmo.o mm.o mfio.o mm.o com: mdo.o #m.o Ndo.o mm.o oao.o m>.o Canvamfln pom :«HonHQ pom CHHUHmwQ umm cflHUHmHQ pom wmwo Hmm pawn Mom ucmo Hmm ucmo mom mo.oH«¢.o mo.ofim¢.o mo.OHN¢.o mo.OHN¢.o .ocoo Hmumkm 803.8 «63.3 8.9.4.3 9034. o 639.3353 .mmusumnmmEmu ucmnmmmwc um uwum3mmu omuwcwnoanomo cw Aammv Newunamwo ou Ummomxm movaosuflaflm mwaflmQqu CH Aunmflm3 um3 .Emmv cwuoamflv mo coHumnucwucoo .wd manms 65 of dieldrin with an especially large decrease (ca. 70%) at 15°C (Table 17). A two-way analysis of variance was run on the means dur— ing the uptake period to determine the significance of the observed differences. A highly significant difference between temperatures and a significance difference with time was found (Table 18). Further investigations using Duncan's (1955) new multiple range test showed that most of the dif- ference was due to the much greater concentrations found at 20°C and the large increase between the second and third weeks accounted for most of the significant change with time (Table 18). In all three experiments (10, 11, and 12) there seemed to be little difference in insecticide uptake at 50 and 100C. In both dieldrin experiments (11, 12) there was a consider- able increase in the concentrations attained at 150C and, where the mussels survived, an even larger increase at 200C. Before any final conclusions are drawn however, tests should be run in natural water to see if temperature influences the uptake and elimination of insecticides the same way when food is present. Conclusions This study has brought out four general conditions which one must consider when using freshwater mussels as monitors of insecticides in water. 66 .ARmmv ucoumMMHv maucmoamwcmwm mum mafia :oEEoo >Q Umcwauwvs: uos mammzm .ARm.mmV ucmnmmmwv wauGMUHMAcmHm mum mafia coEEoo wn Umcwauwocs no: mammzm mow.o m¢H.o mma.o mmcmmz m N a mxmmz >mw.o d0d.0 Nmo.o bmo.o mmcmmz Doom oomd 00m uooa mnsumnmmea dd m¢mm0.0 Hmuoa mmooo.o m dmmoo.o HOHHM fid.m «5.0 ammoo.o N mmhoo.o mafia mm.md mfl.mm «deo.o m thmo.o musumnmmfima mm .m mmmoh omxmm mg mm mm mUHgom .mnsumummEmu 0cm musmomxm mo numcma ou uommmmu nufl3 mmUHommfiawm maawmmqu cw mc0wumuucmosoo saunawwo coma Ga mwoswuwmmwo Um>ummno mo moamuwMHc Imam map How mummy mmcmn mHmHuHDE cam mucmwum> mo mammamcm am no muasmmm .md magma l‘l'lll ll [I It I 'lllfllilll‘ll itf. {I'll [ [(lll‘l ‘(l' 67 First the previous conditioning and insecticide level of the mussel may influence the concentration of the insecti- cide attained by the mussel. This seems to be especially important when they are placed in waters which contain little or no natural food. The mussel's filtering rate is dependent on the quality and quantity of food in the water (Wilbur and Yonge, 1966). This leads to the second consideration, the type of water into which the mussels are placed. As mentioned above, the level of available suspended matter for food affects the filtering rate. A large difference was found in both uptake and elimination in the experiments between conditions of no food (distilled and tapwater) and essentially unlimited food (Lake Lansing water). Thus mussels placed in cold clear streams would probably yield different results from those in streams which had native mussels populations. It is also possible to have a suspended load that is too great and results in decreased filtering (Loosanoff and Engle, 1947). Poor water quality conditions which cause the mussel to close up and not filter for periods of time, may yield lower than representative concentrations of insecticides in the mussel: for the water. This condition apparently occurred in previous field studies (Bedford §£_al., 1968). As would be expected, the temperature of the water is another important consideration. At 10°C and lower the mussels appear to concentrate insecticides at the same low 68 levels but large differences were noted between 10°, 15°, and 20°C, both in rate of uptake and equilibrium concentra- tion. Finally, the type of insecticide being monitored must be considered. Although both DDT and dieldrin are classed as chlorinated hydrocarbon insecticides, the degree to which they were concentrated and their half-life in the mussels were considerably different. Most of these limitations would apply to any organism used for monitoring pesticides. Thus the mussel's traits of feeding by filtering large quantities of water, moving very little, and long life span (up to 20 years) makes them especially well adapted as monitors in comparison with other aquatic organisms. Galtsoff (1928) found that adult oysters, of a size similar to freshwater mussels (5—4 in long) siphoned up to 5,000 ml/hr when the water temperature was 25°C and siphoned, on the average, 20 hr a day at a temperature range of 15-22OC. Bovjerg (1957) reported that the mean movement of Lampsilis siliquoidea when well fed was only 2.5 m per week and when not fed the mean movement ranged from 5.4 to 6.7 m per week. SUMMARY Fresh water mussels were exposed to several concentra- tions of DDT and dieldrin between 0.05 and 1.0 ppb in (reconstituted distilled water, dechlorinated tapwater, and natural lake water under continuous flow and constant temperature conditions. The mussels concentrated DDT approximately 1000 fold in distilled water and 2400 fold in lake water. Dieldrin was concentrated about 1200 fold in lake water by the mussels. The concentration of insecticide in the mussels reached equilibrium with the concentration in the water faster in lake water than distilled water and the insecticide also had a shorter half-life in the mussel in lake water. Dieldrin's half-life was 4.7 days in lake water, about one-third of DDT's half-life in lake water. The insecticide concentrations were highest in the diges- tive and reproductive tissue and low in the muscle, mantle, and gill tissues. The concentrations were very 69 70 low in the marsupia in tests run in distilled water but were almost as great as the digestive and reproductive organs in lake water. Temperature was found to affect the rate of uptake and elimination of the insecticides in dechlorinated tap- water. It also affected the equilibrium concentration in the mussels. LITERATURE CITED LITERATURE CITED Arthur, J. W. and E. N. Leonard. 1970. 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Pesticide monitor- ing of the aquatic biota at Tule Lake National Wildlife Refuge. Pesticides Monit. J. 1(4): 21—26. Grzenda, A. R., D. F. Paris, and W. J. Taylor. 1970. The uptake, metabolism, and elimination of chlorinated residues by goldfish (Carassius auratus) fed a 14C-DDT contaminated diet. Trans. Amer. Fish. Soc. 99(2): 585-596. Li, J. C. R. 1964. Statistical inference I. Edwards Brothers, Ann Arbor, Michigan. 658 pp. Loosanoff, V. L. and J. B. Engle. 1947. Effect of different concentrations of micro-organisms on the feeding of oysters. U. S. Fish & Wildlife Serv. Fish. Bull. 51: Miller, C. W., B- M. Zuckerman, and A. J. Charig. 1966. Water translocation of diazinon—C14 and parathion-Sas off a model cranberry bog and subsequent occurrence in fish and mussels. _Trans. Amer. Fish. Soc. 95: 545-549. Reinert, R. E. 1970. Pesticide concentrations in Great Lakes fish. Pesticides Monit. J. 5(4): 255-240. Robeck, G. G., K. A. Dostal, J. M. Cohen, and J. F. Kreissel. 1965. 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