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I Accumulation of Aroclor 1254- in Grass Shrimp (Valucmonetes puglo) in Laboratory and Field Exposures by D. K. Nimmo, J. Foiu.steh, P, T. Hkitmuller, and G. H. Cook U. S. Environmental Protection Agency Culj lireeze Environmental Research Laboratory Sabine Island, Gulf Breeze, Fla. 32581 Results of several experiments Indicate that aquatic inverte brates accumulate total body concentrations of polychlorinated biphenylB (PCS) thousands of times greater than that of the sur rounding water. For example, SANDERS and CHANDLER (1972) showed that fresh water insects and crustaceans rapidly (1 day) accumu lated PCB (Aroclor 1254) up to 24,000 times greater than the concentration in the water. Results of similar exposures con ducted with estuarine animals showed oysters concentrating 85,000 (LOWE ct si. 1972), shrimp 10,000 (NIMK0 et al. 1971a), and fish 30,000 (HANSEN et ol. 1971) times the amount of PCB in the water. .. Although SANDERS and CHANDLER (1972) stated that PCBs entering the aquatic environment are below concentrations acutely toxic to invertebrates, we have noted that most of the accumu lation studies conducted thus far by the investigators cited in the paragraph above have been at concentrations of 1.0 ug/i- and above, i.e., concentrations demonstratively toxic to test animals. Little is known about accumulation in marine invertebrates at extremely low concentrations, and with one exception (NIMMO et al. 1971a), no one to our knowledge has placed P-CB-free animals in a natural environment known to have PCBs and followed accumulation with time. Ve report here the results of several experiments on chronic toxicity of Aroclor 1254 to Palaetnonetes puglo, an estuarine grass shrimp, as well as concentration and lose of the compound from the animals with time. We also exposed grass shrimp for up to 3 months to Aroclor 1254-contaminated sediments in Escambia Bay, near Pensacola, Florida. METHODS AND MATERIALS With one exception, all laboratory experiments were conducted in 30-ml chambers supplied with flowing water from Santa Rosa Gulf Breeze Environmental Research Laboratory Contribution No. 170 Associate Laboratory of the National Environmental Research Center, Corvallis, Oregon 303 ItulMin iif Ln'Iromiu-nUi C'oimji>fltion A T^mkolofly, Veil. 1},Nm.-1 C JV74 by Nrw Yorfc Inc. yro'-vir: >;/.,, .rr-. a Viv'*: -*'.v t4v- ' A< 1 : #y-.' i "!* i,;Tpr ,},-p : S.} yv* >Is>'y [ Svi S> vz 3-C r A,)-;: ' -aviv'-V- V'--'y-.q-A t it V* ; y- . v;*> I pi 7 * -iV-'"?.; V $ i'Kr: i":.v0y teItbts i I,A; V1' r ' .V v. 1 !*03, ' '.I-,' -1.?-rw*,' '..AT'*' ryi fe- v-v -!;d bi A :;;-T>'A(v|:;Ly A'.;r--. '< 'v ' ^ i.V;V TvV..- DSW 025356 i;- STLCOPCB4009311 v:flujh; :; iK-i^'t/L. U vf Vj- iiCtv*Ji. il Sound. Three sets of 5 chambers each received test concentrations: the fourth set was a control. Each chamber contained 4-10 shrimp, the number depending on the size of the animals. Water flowed continuously through each chamber at a rate of 1,0 1/hr. Aroclor 1254, dissolved in polyethylene glycol (mol, wt. 200), was metered into each mixing tank with a syringe pump before the water entered the test chamber. An equal amount of solvent was added to the water flowing to controls. David J, llansen of this laboratory, found the sensitivity of a marine fish to Aroclor 1254 remained unchanged when he varied concentrations of polyethylene glycol used to deliver the toxicant (personal communication). The shrimp were fed daily a commercial molly-flake diet (<0.02 mg/kg organochlorine compounds). The experiment to determine the concentration and loss from the tissues of JP. puglo was also conducted in a flowing-water system, We constructed 18-liter aquaria with false floors of nylon screen (1/4-inch mesh) to hold shrimp above the detritus brought in with the water or produced by the animals. This modi fication was intended to prevent the animals from eating these particles with adsorbed Aroclor 1254. Consequently, we assume that Bhrtmp obtained more of the chemical from the water by ab sorption through the gills rather than from ingestion of con taminated detritus. The shrimp were not fed during thiB experi ment. Concentrations of Aroclor 1254 in tissues by gas chroma tography were determined uBing pooled samples of at least 10 shrimp each (NIMM0 et al. 1971a). P. pugio were exposed to Aroclor 1254-contaminated sediments in upper Escambia Bay from November 1971 to February 1972. The shrimp in specially-constructed cages (HEI1WULLER and N1MM0, 1972) were exposed directly to the sediments. Average concentration of Aroclor 1254 in the uppermost two inches of eediment in November 1971 was 5.0 mg/kg (dry weight). RESULTS OF LABORATORY EXPOSURES Tests conducted in flowing water showed jP. puglo to be sus ceptible to Aroclor 1254 (Table 1). In a 7-day exposure, 60% died at 9.1 pg/fc, but significant mortality did not occur at 0.17 and 0.62 pg/i.. In the second series of tests lasting 16 days, 4.0 and 12.5 pg/i were toxic, but significant mortality did not occur in 1*3 pg/jt* At the conclusion of several one-week exposures to a range of concentrations (0.17 to 9.1 pg/t), surviving shrimp from each ex posure were analyzed for whole-body residues. Ambient concen tration of toxicant in the water and resultant residues in the shrimp were correlated (r-0.91, Table 2). In some cases, dupli cate test concentrations produced biological accumulations that differed by a factor of 2. Concentration factors ranged from 3.000 to 11,000, These ranges were similar to those found in 301 OSW 025357 STLCOPCB4009312 si:?& TABLE 1. MORTALITY AND ACCUMULATION OP AROCLOR 125*) IN Palaemonetes pugio* Test Cone. Days (hg/) Exposed Average Mortality Body Cone. Concentration (%)** (mg/kg) Factor CONTROL 0.17 0.62 9.1 7 4(0 - 20) 0.1 7 8(0 - 40) 1.3 7 4(0 - 20) 5.4 7 60(20 - 80)*** 65.0 CONTROL 16 25(0 - 50) <0.1 1.3 16 40(0 - 100) 18.0 4.0 16 45(25 - 50)*** 27.0 12.5 16 55(50 - 75)*** 46.0 7600 8700 7100 14000 6700 3700 *A11 exposures were conducted in flowing seawater: salinity and temperature ranges were 22 to 2$* and 17 to 28 C. **5 replicates per concentration: at least 4 shrimp per replication. ***Signlficant at P >C1.05, TABLE 2. ACCUMUIATION OF AROCLOR 1251) BY Palaemonetes pugio* Test Cone. (vg fl) Body Cone. (mg/kg) Concentration Factor 0.17 0.62 1.0 2.3 2.7 3.2 3.2 5*2 5.3 5.3 9.1 1.3 5.4 3.2 25.0 19.0 15.0 26.0 29.0 16.0 30.0 65.0 7600 B700 3200 11000 7000 4800 8100 5600 3000 5700 7100 *7-day exposures conducted In flowing seawater at salinity and temperature ranges of 22 to 28 and 17 to 28 C. 305 :'} T-. r. A^. -- `' OSW 025338 t: ; STLCOPCB4009313  <*>>- V---' .. ' V_ v ': &'"'ci. V <? ACCUMULATION OF AROCLOR 1254 IN Palaemonetes pugio WITH TIME AFTER EXPOSURES TO THE CHEMICAL IN WATER AT THREE CONCENTRATIONS (ug/S.) (Each value represents a composite sample of 10 animals) I', ;A> (t- / ^ > / ' * - ^ ( Length of Exposure (hr/ days) Control Body- Cone. Cone . Factor (mg/kg) 0. 04 Body Cone. Cone, Factor (mg/kg) 0. 09 Body Cone. Cone. Factor (mg/kg) 0. 62 Body Cone. Cone. Factor (mg/kg) ';vV^w,4v* '>' 'T '' . J 0f>`> vV"V :,->:.'.v; -* .*' i\ u'- 0 1 2 3 4 8 12 16 24/1 0.1 --- -- -. --r tt __ -- -- A A A * * * A * * 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 A A A A A A A A A 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 * A A A A A A A A 0.1 0.1 0.1 0.1 0.1 0.12 0.14 0.26 0.20 A A A A A 190 230 420 320 r vl"' -> ' r'" `t : v v : ' >'-v, ..-V It.'- ' ;,' &*5* r, ^',, '?^ i'.:<4irV^:ir..:x-.' K__ > $- * r * * * , * - ' v ?.,. * .. ' * - * 4 > 36 / 1.5 -- A 0.1 A 0.1 A 0.20 470 48/2 72/3 --- * A 0.1 0.1 A A 0.10 0.14 1100 1560 0.37 0.58 600 930 L&. . u s -* A* * ,> .-"j*: *, . . ^ V ' ,. v ' : o 96/4 * 0.1 A 0.15 1670 0.40 650 to x 154 / 6.5 336 / 14 0.1 A A 0.1 1590 0.13 3250 0.33 0.43 3670 4780 1.28 2060 7.40 11930 0 N) 504 / 21 672 / 28 0.1 0.14 A A 0.15 3750 0.17 4250 0.45 5000 1.57 17400 6.67 10900 10.82 17450 Ul OJ 01 840 / 35 0.10 A 0.21 5250 PCB - STOPPED 0.75 8330 16.48 26580 i fe- ' *0 1176 / 49 0.1 A 0.1 A 0.12 A 3.24 A &''. 1512 / 63 0.15 A 0.1 A 0.13 A 1.64 A r;: * Magnification factor not calculated. STLCOPCB4009314 tests using penaeid shrimp (NIMMO et al. 1971a), but were some what lower than those found by SANDERS and CHANDLER (1972), in tests using several invertebrate species in fresh water. There appeared to be no threshold below which levels of the chemical added to water failed to produce residues in the tissues (Table 3) in our tests. Whole-body concentrations produced after 5 weeks exposure to 0.04, 0.09 and 0.62 pg/j, ranged from 200 to 26,000 times the concentrations in the test water. Concentrations did not reach equilibrium and from 60 to 90 percent of the Aroclor 125A was lost from the shrimp within 4 weeks after exposure to the chemical was stopped. Test concentrations of the chemical were not significantly toxic to shrimp. Although accumulation increased with increasing concentration of toxicant in this test, this was not observed in earlier studies (see Tables 1 and 2). Implications are to be discussed elsewhere. RESULTS OF FIELD EXPOSURES Average whole-body residue of Aroclor 1254 in _P, puglo after 1 month was 0.41 mg/kg (0.34 to 0.57); after 3 months, 0.42 mg/kg (0.37 to 0.50). There was no evidence that significant mortality occurred during the exposures of grass shrimp to contaminated sediments. DISCUSSION Concentrations of Aroclor 1254 in pugio, after exposure to contaminated sediments for 3 months was equivalent to a Vu laboratory-exposure of 0.09 pg/j, in water for 2 weeks (Table 3). We expected residues to be higher in caged shrimp since we had found that fiddler crabs exposed in the laboratory accumulated U residues equal to or greater than (wet-weight basis) that of the rO<-t contaminated substratum (dry-weight basis) after 30 days u (N1MM0 et al. 1971b), Concentrations of Aroclor 1254 In caged OV shrimp exposed to contaminated sediments appeared to reach a u4oJ plateau, but this was not the case in laboratory exposures (Table 3) where an equilibrium was not reached. Therefore, <o ve believe that shrimp exposed to the sediments might have obtained PCE from the water or food singly, but shrimp exposed to co Aroclor in the laboratory obtained chemical from two sources, &o water and food. It might also be more available in the laboratory titan the field due to the carrier. In earlier laboratory studies 'i* with penaeid shrimp, both water and food appeared to be sources v< [i (NIMMO et al. 1971a). No significant mortality was observed in caged 6hrimp and none would be predicted since residues produced in the field were similar to those found in shrimp after laboratory exposures which caused no death. Penaeid shrimp spend only a fraction of their life cycle in an estuary, moving into oceanic waters after reaching maturity 307 DSM 025360 STLCOPCB4009315 ' *, 7' I. T- V ' ' '.I,'- \. .'.J'i . P T-V' >'<;? . a, V'-'i.-V:'"-^A.Tv '<v ' > * *. ' > v- ` V' v 7. ''; jz***'------------ ------------------------- miPei^r-av-A*1 t,1, I'i; (PEREZ FARTANTE 1969), but grass shrimp are endemic in estuaries. Therefore, in relation to time of exposure we would expect grass shrimp to accumulate a pollutant from a contaminated estuary to a greater degree than penaeid shrimps, nevertheless, this i6 not true. In August 1968, penaeid shrimps (Penaeus duorarum, , setlferus. and IP. aztecue) collected during a survey of Escambia Bay, Florida, had whole-body residues of Aroclor 1254 as high as 14.0 mg/kg (NIMMO et al. 1971b). In that survey and in subsequent collections, P. pugio had a maximum residue of only 1.4 mg/kg. Lower residues in P. pugio from Escambia Bay may be due to amounts of PCS in bay sediments and behaviorial patterns of the animals. We noted earlier (NIMMO et al. 1971a) In species of penaeid shrimp were related to higher concentrations of Aroclor 1254 in sediments that predominate in upper Escambia Bay. We found that penaeid shrimp, as adults, usually are captured in deeper waters and burrow into silty or sandy substrates. In contrast, grass shrimp usually do not burrow, rather are found along shallow sandy beaches and grass beds, where they obtain food that Is relatively uncontaminated with PCB. PiiisiaKiV'-VW'iJ'.sK-'.'Ir'X,' tenn tv4ct/-,s|Jl7*S4r'fAv-Pi.*(W>.j7v!i*!'.t.Sw`|at.4v }. REFERENCES HANSEN, D. J., P. R. PARRISH, J. I. LOWE, A. J. WILSON, JR., and P. D. WILSON: Bull. Environ. Contam. and Toxicol., 6^, 113 (1971), HEITMULLER, P. T., and D. R. NIMMO: Prog. Fish-Cult., 34, 120 (1972) . LOWE, J. I., P. R. PARRISH, J. M. PATRICK, and J. FORESTER: Mar. Biol. (Berl.), 17, 209 (1972). NIMMO, D. R., R. R. BLACKMAN, A. J. WILSON, JR., and J. FORESTER: Mar. Biol. (Berl.), 11, 191 (1971a). NIMMO, D. R., P. D. WILSON, R. R. BLACKMAN, and A. J. WILSON, JR. Nature, 231, 50 (1971b). PEREZ FAREANTE, X. 461 (1969). U. S. Fish Wlldl. Serv., Fish. Bull., 67, SANDERS, H. 0., and J. H. CHANDLER: Bull. Environ. Contam. and Toxicol., 7, 257 (1972). 308 OSW 025361 TYtf/' SfcUl-'#: ^ - * STLCOPCB4009316