Estuaries Vol. 24, No. 4, p. 585-596 August 2001

Residues of Toxaphene in Finfish and Shellfish from Terry and

Dupree Creeks, Georgia, U.S.A.

KEITH A. MARUYA x,*, TINA L. WALTERS 1, a n d RANDALL O . MANNING 2

1 Shidazvay h~stitute of Oceanog'rap]2y, 10 Ocean Science Circle, Savannah, Geo~g'ia 31411 Georg-ia Depa~t,mer~t of Natural Resources, 205 Butler St'feet, Southeast, Atlanta, Georgia _303_34

ABSTRACT: To better characterize human health risks ~ssociated with potentially contaminated seafood, 56 composi te samples of edible tissue of several filifish and shellfish species were anal)~ed for r~sidu~s of toxaphene using g'~s chromatogTaphy wifll electron capture and negative ion ma.~s spectrometric detection (GC-ECD and GC-ECNI-MS). Tox- aphene in fll~se samples, col lected in 1997 near a fo rmer toxaphene plant in Brunswick, Georgia, were previously reported as non-detectable using non-selective techniques. F~sfimated total toxaphene concentrat ions (~2TOX) ranged from less than 0.01 to 26 Ixg g-~ on a wet tissue basis. Smaller, bot tom dwelling finfish such as croaker, mullet, and spot exhibited the high~st ~2.TOX (0.76-26 Ixg g 1)~ larger predatory fish including seatrout contained intermediate levels (0.08-4.4 Ixg g ~), and shellfish (blue crab and shr imp) contained file lowest levels (< 0.01 to 0.27 Ixg g ~). For a given species , samples f rom the site furthest from file toxaphene plant had lower ETOX than samples f rom the oilier 3 sites. On a congener specific basis, levels ranged f rom < 0.0025 to 3.5 Ixg g i Congener distributions were, in general, domina ted by 2-exo,3-endo,6-exo,8,9,10-hexachlorobornane (Hx-Sed) and 2-endo,3-exo,5-endo,6-exo,8,9,10-heptachlo- robornane (Hp-Sed), breakdown products of Cls-Cll0 toxaphene homologs. Other p rominen t congeners conf i rmed by GC-ECNI-MS included Padar numbers 26, 40/41, 42, 44, 50, 62, and 63, as well as several unidentif ied C16-C10 homologs. Minor dif ferences in congener distribution among spec ies and sampling locafiolts suggested dlat exposure reg-im~s a n d / or intrinsic biotra~tsformation capabilities were not uniform. These results indicate that toxaphene r~sidu~s were de- tectable in all spec ies surveyed and at concentrations higher fllan ~stimated previotLsly.

InU-oducf ion

Toxaphene is the trade name of a complex or- ganochlor ine pesticide formula t ion manufac tured in the United States by Hercules, Inc. at their Brunswick, Georgia facility. Technical grade toxa- phene was p roduced by exhaustive chlor inat ion of camphene , resulting in a complex n-fixture of sev- eral hund r e d C1G-CII0 m o n o t e r p e n e compounds with an average chlorine content of 70% by weight (Saleh 1991). Due to its physicochemical and tox- icological propert ies, toxaphene has the potential to accumulate locally in sed iment (Veith and Lee 1971) and biota (Pollock and Kilgore 1978) and exert toxic effects in the aquatic env i ronment (U.S. Environmenta l Protect ion Agency 1980). Residues of toxaphene are also t ransported over long distances via the a tmosphere (Stanley et al. 1971) and as a result are detected in biota in re- mote regions such as the Arctic (Bidleman et al. 1993). Al though it was banned in the U.S. and oth- er countries during the 1980s, toxaphene contin- ues to be an environmenta l quality issue at local, regional, national, and internat ional levels (Agen- cy for Toxic Substances and Disease Registry 1998;

* Corresponding author: tele: 912/598-2315; fax: 912/'598- 2.310; e-mail: kam@skio.peachnet .edu.

U.S. Environmental Protect ion Agency 1998; de Geus et al. 1999).

In the aquatic envi ronment , in situ processes act to modify the distribution of individual congeners in technical toxaphene . A p r o n o u n c e d shift toward lower chlorinated homologs was first repor ted in reducing salt marsh sediments spiked with toxa- phene (Williams and Bidleman 1978). In sedi- ments f rom lakes collected decades after t rea tment with toxaphene , the two most abundan t toxaphene r e s idue c o n g e n e r s were iden t i f i ed as 2-exo,g- e n d o , G - e x o , 8 , 9 , 1 0 - h e x a c h l o r o b o r n a n e and 2- endo,g-exo,5-endo,G-exo,8,9,10-hep tachloroborna- ne (Stern et al. 1996). These compounds , also known as Hx-Sed and Hp-Sed, respectively, are only very minor componen t s of virgin toxaphene. Exper imental evidence indicated that Hx-Sed and Hp-Sed result f rom dechlor inat ion of C1 s and C19 toxaphene componen t s under reducing condit ions (Fingerling et al. 1996). These weathered, sedi- ment-associated residues of toxaphene are also available to biota, including forage and predator fish (Miskimmin et al. 1995; Maruya and Lee 1998). In heavily contaminated systems (Calero et al. 1993; Maruya et al. 2000), the complex toxa- phene residue pat terns in sediment and tissue are in contrast to samples from remote areas, where the pat tern is characterized by fewer, recalcitrant compounds (Alder and Vieth 1996).

�9 2001 Estuarine Research Federation 585

586 K, A. Maruya et al.

I 0 5 1 0 mlIe

St. Slmon~ I~land

Brunswick

Ir Island

Atlantic Ocean

CIJarlcsron, SC

Brunswlr C,A

Fig. 1. Map of study area, Terry and Dupree Creeks marsh, near the Hercules Inc. i~acility in Brunswick, Georgia. Samples were collected in each of tBur zones (A-D).

Gas chromatography with electron capture de- tection (GC-ECD), the basis for standard protocols such as U.S. Environmental Protect ion Agency (1996) Method 8081, offers good sensitivity and repeatability for o rganochlor ine residues, includ- ing toxaphene. At the same time, however, this me thod slfffers f rom poor selectivit}, and hence, specificity for toxaphene. The utility of GC-ECD as a sole technique for the characterization of toxa- phene residues based on unmodif ied technical mixtures as standards of reference is fur ther com- promised by p r o f o u n d changes in congener pro- files. In recent years, the application of electron cap tu re negat ive ion iza t ion mass s p e c t r o m e t r y (GC-ECNI-MS) and the commercia l availability of pure standards have allowed analysts to better char- acterize toxaphene contaminat ion (Muir and de Boer 1995). Thus, GC-ECNI-MS used in conjunc- tion wifll GC-ECD can help distinguish toxaphene residues from other c o m m o n contaminants (e.g., PCBs, chlordanes, and DDTs).

In 1997, the Georgia Depar tmen t of Natural Re- sources conduc ted a study in the Brunswick area to de termine tile levels of toxaphene in several species of finfish and shellfish consumed by hu- mans (Georgia Depar tmen t of Natural Resources 1997). Such informat ion was needed to better as- sess h u m a n health risks due to toxaphene contam- ination. Specimens of each target species were col- lected from several tidal creek sites near the for- mer toxaphene facility, composited, and analyzed using protocols based on U.S. Environmental Pro- tection Agency (1996) Method 8081. As defined by a consor t ium of regulatory and industry represen- tatives (Carlin et al. 2000), toxaphene in each and every one of these samples was repor ted as not de-

tectable. In this study, we combined GC-ECD with GC-ECNI-MS and re-analyzed these samples to pro- vide more definitive estimates of toxaphene resi- due levels and also to de te rmine the concentra- tions of a suite of individual toxaphene congeners.

Materials and M e t h o d s

8~MMPLE COLLECTION ~MND COMPOSITING

In the spring of 1997, blue crab (Calli~ectes sap- idus), shrimp (PeHa~us sp.), and nine different spe- cies of finfish were collected in four zones (A-D) of the Terry-Dupree Creeks estuarine marsh ileal" the Hercules, Inc. facility in Brunswick, Georgia (Fig. 1). Using gill nets a n d / o r baited wire traps, up to 15 individual f i sh /blue crab per species were collected in each zone, measured, and tagged. To maintain size similarity, the total length of smallest individual was > 75~, of the total length of the largest individual. Shrimp (15-20 individuals; 300- 400 g total) were collected in each zone using cast nets and placed in re-sealable plastic bags. All spec- imens were kept on ice. After re turn to shore, ed- ible tissue consisting of scaled, skin-on filets or blue crab lump and claw meat f rom 2-5 individuals were processed and pooled into 1-3 composite samples per zone for each species (Table 1). For shrimp, heads and exoskeletons were removed. Pooled tissues were then g round using a food pro- cessol; transferred to clean glass jars, and frozen. A total of 56 composite samples were delivered in ice-filled coole rs to the Skidaway Ins t i tu te o f Oceanography for analysis. U p o n receipt, samples were immediately unpacked and placed in a freez- er.

Toxaphene Residues in Terry and Dupree Creeks

TABLE 1. Nmaaber of composite edible tissue samples by species and by location. See Fig. 1 for location of zones.

587

species zone 11o composite samples

Blue crab ( Ccdgi,zectes sapid.~s)

Shrimp (Pe*zaeus sp.)

Atlantic croaker ( Micr@ogo*~,ia, s ,e~**da, kzta, s) Striped mullet ( Mag'il c(/)halas) Red drmn ( Sciae,zops ocdla, tm) Spotted seatrout (@.r~oscior~ .r~egndos.a~s)

Spot ( Leiostom~s xar, th~'e~s)

Yellowtafl ( Bairdidla ch.r~so.a~.ra) Black oh-an (Pogwdas crc,mis) Flounder (Paralichthys sp.) Wtfiting/kingfish (Mer~ticirrhz~s sp.)

A, B, D 3 per zone C 2 A, B 1 per zone C S D 2 B, D 2 per zone A, B, D 3 per zone A, B, D 1 per zone A 3 B 2 C, D 1 per zone A, D S per zone g 1 C 2 A 3 A 1 D 1 D 1

SAIvIP LE PROCESSING

For each sample, a 5 to 20 g aliquot of thawed tissue wa~s homogen ized with ~50 g of kiln fired, anhydrous Na2SO 4 using a solvent-rinsed, ceramic mortar and pestle. This mixture was placed in a glass thimble and extracted with 400 ml CH~CI~ in a glass Soxhlet apparatus for > 16 h. The CHeC12 extract was transferred to a glass a m p o u l e / f l a s k / Snyder co lumn apparatus and reduced to < 10 ml using a heated water bath. The extract was then evaporated to dryness in a fume hood and extract- able lipid de termined gravimetrically to the near- est mg using a microbalance. The sample was re- dissolved in hexane and applied to a 500 (L) • 11 mm (OD) glass co lumn packed with 18.0 g 1.0% water deactivated Florisil (60-100 mesh chromato- graphic grade, Fisher Scientific, Fair Lawn, New Jersey), prepared as described in detail in Maruya and Lee (1998). To minimize break through of lip- id, a max imum of 200 mg extractable lipid was ap- plied to each Florisil column. Two fractions were collected from Florisil, the first fraction (F1) was eluted with 50-80 ml hexane to capture PCBs and other nonpo la r interferences, and the second (F2) was eluted with 150 ml of a 20% (v/v) mixture of CH2C12 and hexane to capture toxaphene residues. Both fractions were reduced and F2 exchanged to hexane using a TurboVap II (Zymark Inc., Hop- kington, Massachusetts). Water bath tempera ture and N 2 (> 99.99%) pressure were maintained at 55~ and 8 psi, respectively. Extracts in hexane were reduced to 1.0 ml and transferred to amber 2-ml glass vials sealed with Teflon-lined silicone rubber septa for subsequent GC analysis.

GAS CHROIVIATOGRAPHY

F1 and F2 extracts (1 p,1) were injected into a Varian g400CX GC-ECD with an 8900 autosampler.

Varian Star c h r o m a t o g r a p h y sof tware (vers ion 4.01) was used to acquire and analyze the chro- matographic runs. A SO (L) • 0.25 m m (OD) fused silica capillary co lumn coated with 0.25 iJom DB-5 was used to separate toxaphene componen t s (JgcW Scientific, Folsom, California). The GC oven was p rog rammed as follows: 120~ (hold ] rain); ramp to 260~ at 2~ rain -1 (hold 15 rain). The injector and detector were maintained isothermal at 270~ and SS0~ respectively. The total run time was 86 rain. To confirm the presence of in- dividual toxaphene analytes, extracts of at least one sample per species were analyzed on a Hewlett Packard 6890 Series II GC coupled to a 5973 mass selective detector operat ing in the ECNI mode. Fused silica columns, S0 (L) • 0.25 mm (OD), coated with either 0.25 iJom of DB-5 or DB-XLB were used for this analysis. Methane at a pressure of ~1 torr was used as the modera t ing gas. The quadruple mass spectrometer (MS) and ion source were maintained at 106~ and 150~ respectively. The MS was scanned between 200-500 daltons at 1.g cycles s 1

Concentra t ions of individual toxaphene conge- ners were de termined by GC-ECD based on a 22- c o m p o n e n t mixture of chlorinated mono te rpenes (T1M2, Dr. Ehrenstorfer, Augsburg, Germany). The 17 ch lo robornane and 5 ch lo rocamphene conge- ners in TM2 represent C10-Cll0 homologs (Table 2). Serial dilutions of TM2 ranging fi-om 2-100 pg in hexane were used to generate a 6-point calibration curve and to compute mean response factors. Con- centrations of several p rominen t congeners not in TM2, including Hx-Sed and Hp-Sed (Table 2) and C17 and C18 compounds whose structures are pres- ently unknoval (Hp-1, Oc-1, Oc-2, and Oc-3; Fig. 2) were estimated using the mean response factor

5 8 8 K.A. Maruya et al.

TABLE 2. Structure, homolog, and cl='omatographic properties of individual dflo:Saqated monoterpene (toxaphene) target analytes. Structm-al formulas m'e for chlorobornanes except for P l l , 19, 15, 95, and 31 whida m-e chlorocamphenes (CC). Relative retention time (RRT) data ~'om SmaKing and Maruya (2001). na = not available.

RP.T RP.T Parlar No Homolog Structure (DB-5) (DB-XLB)

11 6 5,5,6-exo,8,9,10-hexaCC 0.585 0.599 12 6 5-exo,6-endo,8,9,9,10-hexaCC 0.591 0.608 15 6 5-exo,6-en do,7-anti,8,9,10-hexaCC 0.619 0.656 l-Lx-Sed* 6 2-exo,S-en do,6-exo,8,9,10 0.621 0.653 21 7 2,2,5,5,9,10,10 0.661 0.675 Hp-Sed* 7 2-endo,S-exo,5-en do,6-exo,8,9,10 0.662 0.669 25 7 5,5,6-exo,8,9,9,10-heptaCC 0.695 0.707 26 8 2-endo,S-exo,5-en do,6-exo,8,8,10,10 0.708 0.687 81 8 5,5,6-exo,8,8,9,9,10-octaCC 0.748 0.788 32 7 2,2,~endo,6exo,8,9,10 0.748 0.777 38 8 2,2,5,5,9,9,10,10 0.792 0.801 39 8 2,2,3-exo,5-endo,6-exo,8,9,10 0.813 0.812 40 8 2-endo,S-exo,5-endo,6-exo,8,9,10,10 0.824 0.828 41 8 2-exo,3-en do,5-exo,8,9,9,10,10 0.824 0.820 42a/b 8 2,2,5-endo,6-exo,8,8,9,10 (or 8,9,9,10) 0.832 0.839 44 8 2-ex o,5,5,8,9,9,10,10 0.844 0.844 50 9 2-endo,8-exo,5-en do,6-exo,8,8,9,10,10 0.888 0.860 51 8 2,2,5,5,8,9,10,10 0.900 na 56 9 2,2,5-endo,6-exo,8,8,9,10,10 0.945 0.928 58 9 2,2,3-exo,5,5,8,9,10,10 0.959 0.947 59 9 2,2,5-en do,6-exo,8,9,9,10,10 0.968 0.955 62 9 2,2,5,5,8,9,9,10,10 1.000 1.000 63 9 2-exo,S-en do,5-exo,6-exo,8,8,9,10,10 1.016 1.008 69 10 2,2,5,5,6-exo,8,9,9,10,10 1.151 1.127

* Not present in the 22-component TM2 mixture (Etn'enstorfer unpublished data).

for all 22 TM2 componen t s . T h e re ten t ion times for Hx-Sed and Hp-Sed were recorded f rom a so- lution provided by Dr. G. Fingerling (Technical University Munich, Germany) . For congeners that were split between F1 and F2, the concent ra t ion was r epo r t ed as the sun: of the 2 fractions, provid- ing that its p resence was conf i rmed in each frac- tion by GC-ECNI-MS (see also Quality Assu rance / Quality Control) .

Total t oxaphene concent ra t ion (ETOX) was es- t imated by calibrating the GC-ECD with a technical t oxaphene p roduc t s tandard (TTX) provided byJ. H o f f m a n (Hercules Inc., Wilmington, Delaware). A known mass of T T X was diluted in CHeC12 to create a concent ra ted stock (~2,800 pog ml 1). Se- rial dilutions were then created in hexane at con- centrat ions between 0.28-55 p~g m1-1, An average response factor for T T X was com pu t ed by sun:- ruing the areas of all peaks in the unresoNed h u m p of unmodi f ied t oxaphene (Fig. 2), and dMding by the kno~m standard mass. T h e T T X response was then appl ied to the s u m m e d a rea of peaks eluting within a specified re ten t ion t ime window (25-60 rain; Fig. 2). For samples with detectable toxa- p h e n e residues in bo th F1 and F2 extracts using GC-ECNI-IMS (see also Qual i t T ~Yssurance/Qualit T Control ) , s was repor ted as the stun of bo th fractions. Areas for peaks co r respond ing to non- t oxaphene c o m p o u n d s eluting within this t ime

window (e.g., C14-C17 PCBs in F1 extracts, and o> ganoch lo r ine pesticides such as chlordanes and DDTs in F2 extracts) were subtracted f rom esti- mates of ETOX. Reten t ion times for PCB and pes- ticide analytes subtracted in this fashion were re- c o r d e d f r o m s t a n d a r d r e f e r e n c e m i x t u r e s SRM2262 and 2261, respectively (National Institute of Standm-ds and Technology, Gai thersburg, Mary- land).

QUALITY ASSUIGM'qCE/QUALITY CONTROL

I n d M d u a l congene r and total t oxaphene con- centrat ions were validated against a c o m p r e h e n - sive, p e r f o r m a n c e based set of quality a s su rance / qua l i ty c o n t r o l ( Q A / Q C ) cr i te r ia . P r o c e d u r a l blanks and toxaphene spiked re fe rence tissue were analyzed in conjunct ion with the samples. Surro- gate c o m p o u n d s (d ib romooc ta f luo rob ipheny l or DBOFB and ot-HCH) were added to each sample pr ior to extract ion to track target analyte recovery. Initial GC-ECD calibration w-ith the TM2 s tandard resulted in r 2 values exceeding 0.99 for all 22 com- ponents . Cont inu ing ins t rumen t calibration was mon i to red using mid-level TM2 and T T X solutions injected every 10-19 samples; the mean relative pe rcen t deviation fro::: the initial calibration re- sponse for TM2 analytes was main ta ined at or be- low 15% for the dura t ion of the study. Mass cali-

(a ) Spot ( A B 6 B 7 B 7 , Zone D)

0.20 - Hx-sed ii

I O . 1 5 f f

0 .10 �9

Hp-sed

0 .05 -

0 . 0 0 ' "

20 . . . . 2 ' 5 ' ' ' 30 ' 35 40

(b ) Technical Toxaphene

0.40 ,

0.30 L

0 . 2 0 :

0 . 1 0 -

ETOX Window: 2 5 - 60 min

;L , . , ',, , . �9

0 . 0 0 : - - , - : ' - - - - - ' . . . . . ' ~ " ' ' "

Hp-1 i

P-40/41/44/42

I . !i :1 ':!!d~!t ~ i : i P-S0/oc-~z,-3

, i !!i ,iii , ii,,

45 so s5 60 ' i ~ " ) o . . . . 7'5

i I

i .iiiii;li'. ~i'~'i~'"k,fi~.>~ ' . ,

;to 25 30 35 40 45 50 55 60 65 70 7B Retent ion Time (min)

Fig. 2. GC-ECD F2 cl=-omatogram of (a) edible tissue of spot (Leiostom%s xar~th%~us) fl'om Dupree Cceek (zone D) and (b) unmodified teclmical toxaphene. Note the predominance of Hx-Sed and Hp-Sed and a shift toward earlier eluEng peaks m (a) relative to (b).

bration of the GC-MS ,~ystem used for analyte con- f irmation was pe r fo rmed weekly.

Five procedura l blanks consisting of ~50 g kiln fired Na.2SO 4 were processed and analyzed using the p rocedures followed for tissue samples. No tar- get toxaphene congeners were detectable by GC- ECD or GC-ECNI-MS in any blank sample; ECD- based E T O X in these blanks ranged between 2.2 to 9.6 ng g 1 (x _+ cr: 6.76 _+ 2.89 ng g 1). Since toxaphene levels in most of our samples were in the p.g g-i range, E T O X were not correc ted for blank levels. Mean surrogate recoveries (x -+ cr) for DBOFB mad oL-HCH were 92 -+ 18% and 79 _+ 19%,

Toxaphene Residues in Terry and Dupree Creeks 5 8 9

respectively. The mean recoveries of TM2 compo- nents and TTX spiked into reference seatrout mus- cle were 107 _+ 11% and 94?o, respectively. Twelve of the 29 TM2 componen t s eluted in F2, 5 eluted in F1, and the remaining 5 were split between F1 and F2.

The presence of a toxaphene congener was con- f i rmed using GC-ECNI-MS if the re ten t ion time (+ 0.1 s) and mass spect rum ( e 70?6) matched that of a target analyte in TM2 (Table 2). Nine individ- ual congeners , Hx-Sed and Hp-Sed, P-26, P-40, P- 41, P-42, P-50, P-62, and P-6S, were conf i rmed in the majority of samples (Fig. 3). Since [M-C1]- fi-agment ions are typically p redominan t for toxa- phene congeners using ECNI-MS (Jansson and Wideqvist 198S), the degree of chlor inat ion for several unident i f ied toxaphene congeners (e.g., Hx-1, Hp-1, Hp-2, and Hp-3; Oc-1, Oc-2, and Oc- 3) wa~s also assigned. Several Clv-Cll0 PCB conge- ners, the majority of which eluted outside the 25- 60 min NTOX re tent ion time window, were con- f i rmed in the majority of F1 extracts. These PCBs were at t r ibuted to Aroclor 1268 contaminat ion originating from a different tidal creek system in the Turtle-Brunswick river estuary, approximately 20 miles away by boat f rom our study site (Maruya and Lee 1998). Peaks cor responding to PCB con- geners in Aroclor 1268 that eluted within the E T O X re tent ion time ~dndow (e.g., IUPAC num- bers 155, 187, 202, and 201) were not included in estimated E;TOX calculations. Pesticide analytes that were potential in ter ferences in our toxaphene congener specific analyses (e.g., oxychlordane, 4,4'-DDE, 4,4'-DDT) were no t detected in our sam- ples using GC-ECC',II or electron impact MS.

DATA AND NTATISTICL, kL ANALYSES

All ins t rument calibration and sample concen- tration data were compiled and analyzed using Mi- crosoft Excel 97 SR-2 spreadsheet software. Statis- tical evaluation of the data using analysis of vari- ance (ANOVA) and t-tests were pe r fo rmed using the data analysis tool in Excel.

R e s u l t s a n d D i s c u s s i o n

TOTAL TOXAPHENE CONCENTRATIONS

Total toxaphene concentra t ions on a wet weight basis (ETOX) ranged from 1.7 ng g-i for blue crab (zone A) to 26,000 ng g-i for mullet (zone B) (Ta- ble .g), a range spanning over 4 orders of magni- tude. Normal ized to extractable lipid content , total toxaphene concentrations (~TOXnp) ranged from 0.59 pog g a for blue crab (zone A) to 270 pog g a for mullet (zone B); lipid normal izat ion reduced the variability in E T O X to a factor of ~-~500. Using linear regression analysis, E T O X and extractable lipid content were positively correlated (~;TOX

5 9 0 K . A . M a r u y a e t a l .

~lln~ TIC: A ~ 7e.72 D

t HxSed

150QC( I Hp-8~l

; ' ! Hp-1 Hp.2 I , 44

51)000" Hx-1 ] i CL7 C~ C~S ~J I I' I II 4Zr

i ~ > 151101551016(1016501700175018.1731850t9001~ ~ m O 0 20 ~0210021 50 22c0225023.00 23.5024.C024 50 25.0025.5026.1xJ26Eo 27.C027,~ 28.~ ~ , ~ ~ . ~

7 379 2,~,xo 5,5,8,9,9,10,10-oclachkxoboma~ 1P..441

:ia~ i a?5 ] 2 1000:

re;z-> 352 354 356 3 5 8 360 3 6 2 364 31~ 31~ 370 372 374 3.76

i a61

'~g"~ad * ';;i"' 'ad'"'ad'"aa;' #d"' '~4"a;;' ad'' "~d'; 'd~a~"; 'd' ~ ' " ~ ' ig ' ';i i"i

Fig. 3. GC-ECNI-MS (top panel ) total ion d ] r o m a t o g r a m of spot (Leiosto.m.~s xanthur.~s) (zone D) and (bot tom panel ) mass spect~-al conf i rmat ion of Parlar 44, a Ca8 c o m p o n e n t of technical t o x a p h e n e (CTT). Also p r o m i n e n t are o ther known CTTs (e.g., P40/41 , 50, and 63), breakdown p roduc t s (Hx-Sed and Hp-Sed), a n d several un iden t i f i ed C16-Caa b o r n a n e res idues (e.g., Hx-1 and Hp-1).

(ng g 1 wet weight) 59S X % lipid + 45S; n 56; r 2 0,82; p < 0 ,00l) ind ica t ing that lipid con- tent is an i m p o r t a n t co-varying factor. Because ~ ;TOX and ETOXI> were also s t rongly cor re la ted (s p (in pog g-f ) 0,0082 • s + 20,9; n = 56; r 2 = 0.72; p < 0.001), it can be in fe r r ed that

any t rends n o t e d for E T O X may be gene ra l i zed to a lipid basis as well.

Species Diffbences T h e 13 samples with the h ighes t E T O X were

compos i t e s o f mul le t , spot, and c roaker (Table 3).

TABLE ..% Ranking of samples according to s (ng g-1 wet weight) in descend ing order.

Species Zone % Lipid ETOX Species Zone % Lipid ETOX

Mullet B 11 26,000 sea t rout B 2.8 940 Spot D 13 23,000 yellowtail A 4.3 840 Mullet B 6.3 17,000 spot A 19 760 Spot B 15 16,000 red ch'um D 0.55 490 Spot D 9.6 13,000 yellowtail A 2.1 370 Spot C 14 12,000 b lue crab D 0.60 270 Ca-oaker B 14 11,000 yellowtail A 1.5 230 Ca-oaker D 12 11,000 sly-imp D 0.69 210 Mullet B 8.2 8,300 s l m m p C 0.99 150 Spot D 12 7,600 sly-imp g 1.1 120 Ca-oaker B 10 6,300 sea t rout A 1.9 110 Spot C 13 5,700 b lue crab g 0.40 95 Croaker D 7.2 5,300 s l m m p C 0.93 93 Seatrout C 6.7 4,400 b lue crab C 0.88 93 Mullet A 11 4,100 s l~ imp D 0.53 91 Seatrout g 6.9 3,600 f l ounde r D 0.24 90 Sseatrout D 2.3 3,300 red oh'urn A 0.93 79 Spot A 16 2,400 b lue crab D 0.56 75 Mullet D 2.1 2,100 s l m m p C 0.72 72 Whi t ing D 3.4 2,100 b lue crab C 1.0 66 Mullet D 2.2 2,000 b lue crab B 0.31 61 Mullet A 4.2 1,800 black d r a m A 1.5 51 Red diqam B 1.7 1,400 b lue crab D 0.43 49 Mullet D 2.6 1,300 b lue crab A 1.1 44 Spot A 21 1,300 b lue crab B 0.50 36 Seatrout A 12 1 ,S00 sh r imp A 1.5 <12 Seatrout A 6.2 1,300 b lue crab A 1.0 6.3 Mullet A S.0 970 b lue crab A 0.29 1.7

Toxaphene Residues in Terry and Dupree Creeks

TABLE 4. Species differences in ~ T O X were s ignif icant us ing one-~.~,ay ANOVA.

591

Groups Count Sum .N~e r age Variance

Blue crab 11 798 78 5,221 Mullet 9 63,570 7,063 76,995,600 Seat rout 7 14,950 2,186 2,592,795 Shr imp 7 748 107 3,892 Spot 9 81,760 9,084 56,691,678

.~3.,IovA Source SS df MS F p value F crit

Between groups 6.15 • 10 a 4 1.54 • 10 a 5.38250 0.002 2.61899 Within groups 1.09 x 109 38 28,556,067 Total 1.7 X 109 42

Conversely, 8 of the 11 samples with the lowest s were blue crab. Roughly half of all samples (27 of 56) had s > 1,000 ng g-1. Overall spe- cies differences in m e a n ~ T O X were observed us- ing one-way ANOVA (Table 4). Of the 5 species with 7 or m o r e composi te samples (blue crab, mul- let, seatrout, shr imp, and spot) , spot had the high- est mean levels of t oxaphene (9,100 ng g 1) and blue crab the lowest (72.6 ng g 1). Using 2-tailed t-tests assuming unequal variances, significant (p - 0.05) differences in m e a n NTOX were noted for blue crab and each of the S finfish species (mullet , seatrout , and spot) , sh r imp and each of the S fin- fish, and sea t rout and spot. No differences in mean ~ T O X were noted for blue crab and shr imp, mul- let and spot, or mul le t and seatrout.

Zone (Sa'mpling Location) Differences Since t oxaphene levels were previously deter-

mined to be species dependen t , s g rouped by species were analyzed by one-way ~ O V A to as- sess for differences a m o n g the 4 sampl ing zones. This was possible for g species (blue crab, mullet, and spot) since s values for a m i n i m u m of 8 replicate samples were needed for at least 2 differ- ent zones (Table 5). For blue crab, no dif ference in m e a n ~ T O X a m o n g zones A, B, and D was ob- served (p - 0.24). For mullet , however, the mean ~ T O X for zone B (17,000 ng g-1) was significantly

h igher than m e a n E T O X for zones A or D (2,300 and 1,800 ng g-l , respectively). For spot, the mean s for zone D wa,s h igher than for zone A (15,000 versus 1,500 ng g- l ) . For a given species, s for zones B (Terry Creek at bridge) and D (Dupree Creek) were, in general , elevated relative to zone A (Terry Creek at Lanier Basin).

An insufficient n u m b e r of samples p rec luded statistical compar i sons for all species in zone C, and for blue crab and shr imp (all zones). T h e m e a n s for zone C spot (8,900 + S,150 ng g 1, n - 2) was however, in te rmedia te between m e a n levels for zones D and A. For blue crab, m e a n N T O X decreased f rom zone D (130 _+ 99 ng g-i) to zone A (17 + 19 ng g-1) with in te rmedia te values for zones B (64 + 24 ng g-l) and C (80 + 14 ng g 1). Similarly, m e a n s decreased for shr imp fi-om zone D (150 _+ 60 ng g 1) th rough zones B and C (120 ng g 1, n - 1; 105 _+ $3 ng g-1) to zone A ( < 12 ng g-1, n - 1). Clearly, biota f rom zones D (i.e., Dupree Creek) , B, and C (lower Ter ry Creek near the Torres Causeway Bridge) conta ined h igher t oxaphene levels than those col- lected fi-om zone A (i.e., the Lanier Basin area). This decreasing t rend in ~ T O X for all species strongly suggests a spatial t oxaphene contamina- tion gradient away f rom the Hercules , Inc. dis- charge canal in Dupree Creek (Fig. 1).

TABLE 5. t, test compar i sons indica te s ignif icant differences in ~ T O X a m o n g sampl ing zones for mu l l e t and spot.

Mullet ,Spot

Zone D Zone B Zone A Zone D Zone A

Mean 1,800 Varian ce 190,000 Observat ions S Hypothesis 0 df 2 t star 2.9906 P(T --< t) 1-tail 0.05 t cri t 1-tail 2.9200 P(T --< t) 2-tail 0.0960 t cri t 2-tail 4.3027

17,100 2,290 14,588 1,487 78,330,000 2,629,300 61,053,333 698,533

3 S 3 3 0 0 2 2 2.8509 2.8756 0.05 0.05 2.9200 2.9200 0.1042 0.1026 4.3027 4.3027

592 K.A. Maruya et al.

TABLE 6. Individual congener concenU'ations (~ + ~ in units of ng g-:) for zone D finfish and shellfish. Value in parentheses are ls.

Mullet (n = 3) Spot (n = 3) Seatrout (n = 1) Blue crab (n = 3) Shrimp (n = 2)

I-Lx-Sed 122 (16.7) 1,420 (8.33) 331 4.12 (0.48) 10.2 (3.15) Hp-Sed 800 (64.9) 1,180 (7,85) 502 2.52 (1.28) 7.42 (2.14) P-26/Hp-1 61.6 (26.0) 549 (226) 129 7.54 (6.30) 6.86 (2.84) P-40/41 16.0 (3.91) 136 (59.5) 40.2 0.78 (0.43) 0.91 (0.28) P-42 16.3 (8.96) 94.4 (36.1) 26..8 <1.29 0.50 (0.22) P-44 27.8 (25.2) 355 (147) 100 1.50 (1.00) -<2.58 P-50/Oc-1 27.7 (2.42) 178 (72.9) 46.2 2.41 (1.17) 4.21 (1.75) P-62 11.4 (2.24) 51..8 (26.9) 17.8 0.86 (0.04) 1.00 (0..39) ]~TOX 1,800 (.860) 15,000 (6,400) .8,.800 180 (98) 150 (60)

TOXAPHENE CONGENER CONCENTRATIONS AND

PROfiLES

T h e concent ra t ions of 98 t oxaphene congeners (or congene r pairs) on a wet weight basis r anged f rom nonde tec tab le (< 0.25 ng g :) to a m a x i m u m of 8,450 ng g- : for Hp-Sed (mullet , zone B). O f the 9 congeners conf i rmed by GC-EGNI-IVIS, Hx- Sed and Hp-Sed were, in general , the most abun- dant followed by (in order of decreasing abun- dance) P-96/Hp-1, 44, 50/Oc-1 , 40/41, 49, 69, and 68. In zone D samples for example , m e a n concen- trations ranged f rom 0.78 ng g : for P-40/41 in blue crab to 1,490 ng g- i for Hx-Sed in spot (Table 6). O n e except ion was no ted for blue crab, where P-96/Hp-1 was m o r e a b u n d a n t than ei ther Hx-Sed or Hp-Sed.

T h e distr ibution of conf i rmed toxaphene con- geners was, in general , similar for all samples (Fig. 4). In previous studies at this site, Hx-Sed and Hp- Sed were also the most a b u n d a n t t oxaphene con- geners in sed iment and biota (Vetter and Maruya 9000). Because these congeners are only mino r c o m p o n e n t s of unmodi f i ed technical toxaphene , they are widely believed to be dead-end metabo- lites of reduct ive dechlor inat ion in con tamina ted

III mullet (n=9) 1.2 . . . . . . . . . . . . . . . . . . . .

~ ..................... P shr imp (n=7) "~ 0.8 ! [] blue crab (n=l 1) I

n- ~ 0.2

0 . 0 . . . .

~0 a: o 0

Congener

Fig. 4. Mean congener distributions illustrate differences in exposm'e regime and/or biou-ansfonnation capability among fmfish and shellfish spedes. Error bars represent +lcr.

anoxic sed iments {Stern et al. 1996). The i r pre- d o m i n a n c e has also been observed in native rain- bow trout f rom toxaphene- t rea ted lakes (Miskim- rain et al. 1995). Several C1 s congeners (e.g., P-40, P-41, P-42, and P-44) were also a b u n d a n t in most of our samples at levels as high as 7SS ng g- : (p_ 44 in mullet , zone B). Relatively few C19 homologs were detected; the 9 congeners that were promi- nen t (P-62 and P-68) were presen t at lower levels ( m a x i m u m of 175 ng g- i in mullet , AB68868). No Cl:0 c o m p o u n d s were detected.

Several unident i f ied Cl~-C19 toxaphene residue c o m p o u n d s were also detected in many of these samples (Figs. 2 and g). P e n t a c h l o r o b o r n a n e s and h e x a c h l o r o b o r n a n e s have been previously identi- fied in Ter ry and Dupree Creeks sediments (Ma- ruya et al. 2000) and biota (Maruya unpubl i shed data) as have several hep tach lo robornanes , includ- ing Hp-1 (Vetter and Maruya 2000). A series of C1 s c o m p o u n d s eluting after P-44, designated Oc-1, Oc-2, and Oc-B, were also p r o m i n e n t in the major- ity of our samples. Al though their s t ructures re- main unknown, they likely occur in technical tox- a p h e n e in relatively high abundances . A compar i - son of indMdua l congene r concent ra t ions and ]~TOX estimates indicate that, in many cases, only a small fract ion ( < ,85%) of the t oxaphene con- geners in our samples have been positively identi- fied (Table 6).

Species and Zone Effects A difference in congene r distribution was no ted

between fish and crustacean species (Fig. 4). Spe- cifically, sh r imp and blue crab had relatively less Hp-Sed and m o r e P-26/Hp-1 and P-50/Oc-1 than fish. C o n g e n e r profiles for shr imp and blue crab were similar, suggesting a similar exposure reg ime a n d / o r e l im ina t ion /me tabo l i c capability. These crustacean species are highly mobi le and feed on planktonic organisms as well as detritus. Profile dif- ferences a m o n g the various species of fish ap- peared to be l imited to Hx-Sed and Hp-Sed (Fig. 4).

Little is known concern ing the metabol i sm of

"O

- , 0.6

0.4 i . >

0.2 n-

0.0 "

~1168785 (zone D) [ ] 68863 (zone B) [] 70215 (zone A)

e q

-1- -I- e , l

Congener Fig. 5. T h e sm~ilm-ity of c o n g e n e r dis tr ibut ions in mul le t

f rom dif ferent sampl ing locat ions sugges t a u n i f o r m exposure reg ime a n d / o r a h igh, local mobili ty for fl~is species.

t oxaphene in finfish and shellfish, with the avail- able data suggesting highly variable biotransfor- mar ion capabilities a m o n g species (Saleh 1991). Recent studies indicate that the e l iminat ion of Hx- Sed and Hp-Sed by fish is relatively rapid with re- spect to P-26 and P-50, two congeners considered to be a m o n g the most recalci trant (Fisk et al. 1998; Smalling et al. 2000). T h e enantioselective na tu re of the e l iminat ion process strongly suggests that these fish actively metabol ize Hx-Sed and Hp-Sed (Smalling et al. 9000). If one assumes that all spe- cies in this study similarly el iminate t oxaphene res- idues and that Hx-Sed is e l iminated faster than Hp- Sed (Smalling et al. 2000), the p r e d o m i n a n c e of Hx-Sed in spot, croaker, and the 2 crustacean spe- cies may indicate un in t e r rup t ed exposure to sourc- es of t oxaphene contaminat ion . This may be due to exposure to sources enr iched with Hx-Sed, e.g., tidal creek water at the site (Maruya unpubl i shed data). T h e p r e d o m i n a n c e of Hp-Sed in species such as mul le t and sea t rout suggests in te rmi t ten t exposure a n d / o r exposure to Hp-Sed enr iched media. Mullet are known to ingest sed iment in search of food, they in turn are preyed u p o n by seatrout.

Spatial differences in t oxaphene profiles by spe- cies were not as obvious. C o n g e n e r distributions for mul le t (Fig. 5) and sea t rout (not shown) f rom zones A, B, and D were similar, suggesting that ex- posure reg imes were un i fo rm th roughou t the study area a n d / o r that fish moved freely a m o n g study zones. Location-specific profiles for shr imp (Fig. 6) and blue crab (not sho'~m) were m o r e var- iable, with congene r complexi ty decreasing f rom zone D (Dupree Creek) to zone A (Terry Creek at Lanier Basin). Because of their lower hydropho- bicity, Hx-Sed and, to a lesser extent, Hp-Sed are

Toxaphene Residues in Terry and Dupree Creeks 593

m o c

"o r , .,,I

a)

0.8 t11![I N ...... tl ......................... []68837 (zone C)'~ i!{t I II 1"-168936 (zone B)'

0"6 liJ~ ~l ~ ..N'(-~176 A) I o . , . . . . . . . . . . . . . .

0 . 0 ' , - - . . . . . .

Congener Fig. 6. The difference in c o n g e n e r distr ibutions for sl~-imp

f rom dif ferent sampl ing locat ions suggests a spatially varying c o n t a m i n a n t exposm'e regm]e for this species. T h e predomi- n a n c e of Hx-Sed may indicate preferent ia l up take of t oxaphene res idues via the water co lumn.

m o r e efficiently t ranspor ted f rom the zones of heaviest t oxaphene con tamina t ion (i.e., zones B- D). It follows that exposure via water absorp t ion may be m o r e impor t an t for the crustacean species. Caut ion should be exercised when in terpre t ing blue crab and shr imp profiles as the concentra- tions of individual congeners in most of these sam- ples were relatively lo~:

COMPARISONS WITH O T H E R STUDIES

Total and individual t oxaphene congene r con- centrat ions in our samples were compared with lev- els for various aquatic species f rom other regions compi led by de Geus et al. (1999) and Saleh (1991). E T O X in our samples were as h igh as max- imurn levels r epor ted for con tamina ted fish eating birds, and 20 times h igher than those r epo r t ed for o ther mar ine fish species. Only b lubber of white beaked dolphins f rom Newfound land had higher t oxaphene levels (88-46 IJ,g g- l ) . s for bot- tom dwelling, forage species in our study were four fold h igher than Lake Super ior lake t rout and m o r e than g0 times h igher than ra inbow smelt f rom Lakes Michigan and Ontar io . Average toxa- p h e n e in carp, a bo t tom dwelling fish, f rom the Grea t Lakes was repor ted to be g.g IJ, g g 1, roughly a third of the mean s for spot f rom Dupree Creek (zone D, 9.1 pog g-X). T o x a p h e n e levels for spot ted sea t rout in our study were comparab le to those for Grea t Lakes salmonids. Very little toxa- p h e n e data is available for invertebrates; concen- trations repor ted previously were typically low (< 0.005) I Jog g-X).

M a x i m u m concent ra t ions for Hx-Sed and Hp- Sed (up to 3.4 9og g- i wet weight) in our samples were two or m o r e orders of magn i tude h igher than

5 9 4 K.A. Maruya et al.

previously repor ted for an}, t oxaphene congener. The majority of congener specific data available are for P-26 and P-50, as the}, comprise a large per- centage of the total toxaphene detected in biota from intensely studied regions, e.g., the nor th At- lantic and Arctic Oceans. Miskimmin et al. (1995) repor ted max imum total toxaphene levels of ~-~0.1 I Jog g-1 in Chatwin and Peanut Lake rainbow trout where Hx-Sed and Hp-Sed were also p redominan t , but did not give estimates of indMdual congener concentrat ions.

T o x a p h e n e c o n c e n t r a t i o n s and d is t r ibu t ions similar to those repor ted herein were no ted for o t h e r species co l l ec ted i n d e p e n d e n t l y d u r i n g 1997-1998 from Terry and Dupree Creeks, includ- ing grass shrimp (Palaemor~etes sp.) and mummi- chogs (Fundulus sp.) (Vetter and Maruya 2000). In contrast, ]~TOX in this study were, in general , two to three orders of magni tude h igher than in simi- lar species caught in o ther areas of the Turtle- Brunswick River estuary and St. Simons Sound (Maruya and Lee 1998). Estimated toxaphene con- centrat ions based on packed column GC-ECD in shrimp (Penaet~s sp.) tails, anchoret (Anchoa sp.), and star d rum (SteEiJbsp.) collected at our site in the early 1970s ranged h o m 0.6-2.5, 8.6-20, and 1.1-8.2 IJ~g g-1 wet weight, respectively (Reimhold and Durant 1974).

IMPLIC&TIONS FOR EGOLOGIr&M~ AND HUI~s HEALTH EFFECTS

E T O X in our samples ranged fi-om < 0.05 to ~26 IJ~g g 1 estimates that are in many cases con- siderably higher than originally repor ted using an- alytical methods based on U.S. Environmenta l Pro- tection Agency (1996) Method 8081 (Georgia De- pa r tmen t of Natural Resources 1997) and specific criteria defining the presence of toxaphene (Car- lin et al. 2000). This discrepancy is not surprising as toxaphene was detected using the criteria of Carlin et al. (9000) if a min imum of four GC peaks in the back half of the ECD chromatogram of un- modified technical toxaphene (e.g., TTX; Fig. 2) were de te rmined to be present in a sample. These peaks were quantif ied as toxaphene if and only if their relative propor t ions in the sample matched that of TTX. The demarcat ion of the back half of the ECD chromatogram is the highest responding peak in technical toxaphene , seen in Fig. 2 as elut- ing between 55 and 60 min. It is clear that selective weathering a n d / o r b io t ransformat ion of technical toxaphene in reducing sedimentary envi ronments results in a highly modified, left-shifted residue pat tern in sediment (Williams and Bidleman 1978; Stern et al. 1996) and in local food web organisms including fish (Miskimmin et al. 1995; Maruya et al. 9000). It is evident f rom the similar, left-shifted

pat terns in our composite samples (Fig. 2) that the criteria def ined by Carlin et al. (2000) were not optimized for detecting residues resulting from technical toxaphene (weathered toxaphene) .

Ano the r impor tan t difference was the pre-sepa- ration of nonpo la r compounds , including PCBs, from toxaphene residues using solid-liquid chro- matography in our stud),. Even though we did not achieve complete separation using Florisil, the bulk of the total PCB mass in our samples eluted in F1 whereas toxaphene residues eluted primarily in F2. This was conf i rmed using G(bECNI-MS. Nei- ther pre-separation of PCBs n o r GC-MS confirma- tion of toxaphene was pe r fo rmed using the 8081- based me thod (Carlin et al. 2000). Fortuitously, the PCB congeners p rominen t in our samples are late e h t i n g Clr-Cll0 homologs, whose origins have been previously linked to Aroclor 1268 (Kannan et al. 1997; Maruya and Lee 1998). Th e potential for PCB in te r fe rence using our E T O X me thod (25-60 rain window) is lessened whereas the potential for in te r fe rence by these same PCBs in selecting and identifying back half toxaphene componen ts using the criteria of Carlin et al. (2000) is increased. Im- proved cleanup (e.g., Iga-ock et al. 1997) and ad- ditional conf i rmatory techniques are needed to bet ter characterize weathered toxaphene residues. Our work represents the first application of chro- matographic cleanup and GC-MS in characterizing the complex weathered toxaphene profile ob- served in these samples.

Both species- and location-specific trends in tox- aphene levels were also observed. Bot tom feeding finfish such as spot, croaker, and mullet exhibited the highest levels, whereas the lowest concentra- tions were consistently found in the two shellfish species (blue crab and shrimp). Larger, predator}, finfish such as seatrout and red d rum had inter- mediate contaminat ion levels ranging from < 0.1 to ~5 p,g g 1 Spatial differences in contaminat ion were also apparen t with tissue levels elevated at sites closest to the original source of toxaphene contaminat ion. Subtle differences in congener dis- tr ibution among species may be the result of vary- ing exposure regimes (e.g., uptake mechanism, time spent in the contaminated creek habitats) a n d / o r intrinsic e l imina t ion /metabol ic capabili- ties.

Science-based regulatory action levels and risk informat ion in general for weathered toxaphene are extremely limited (Agency for Toxic Substanc- es and Disease Registry 1998). Translat ion of our results into ecological and h u m an health risks is typically based on extrapolat ions to unmodif ied technical toxaphene. Recent investigations indi- cate that weathered toxaphene residues at sub ppm (= pog g-l) tissue levels may affect reproduc-

tion and possibly induce DNA damage in grass shr imp (Palaemonetes sp.), an impor t an t species in salt marsh ecosystems (Lee and Maruya 1999). O the r exper imen t s suggest that Fundulus are less susceptible to t oxaphene con tamina t ion based on a bat tery of bo th acute and chronic toxicological endpoin t s (Maruya and Lee unpubl i shed data).

It was recently d o c u m e n t e d that Fu, ndu, lus can rapidly el iminate some residues of toxaphene , in- cluding Hx-Sed and Hp-Sed, the two p r e d o m i n a n t congeners found in the Ter ry-Dupree Creeks eco- system (Smalling et al, 9000; Vetter and Maruya 9000). O t h e r congeners such as P-26, P-50, and B7- 1000, a recently identified C1 v b o r n a n e isolated f rom Dupree Creek sediments (Vetter et al, 2001), were m u c h m o r e recalcitrant, These congeners ap- pear to be resistant to metabol i sm in other species (e.g., mar ine and terrestrial mammals ; Boon et al. 1998) a~s well as in fish (De lo rme et al, 1988; Fisk et al. 1998). It follows that the m o r e labile toxa- p h e n e residues in biota f rom our study a rea will be removed via e l imina t ion /me tabo l i c processes (natural a t tenuat ion) if con tamina t ion sources are reduced a n d / o r el iminated. T h e long- term ecolog- ical and h u m a n heal th risks fi-om toxaphene in this system can thus be expec ted to shift to recalci trant congeners ,

ACKNO1,gLED GMENTS

The au thors thank K. Smalling, L. H a r m W. Vetter, G. Fin- gerl ing, a n d L. Francendese . This work was suppor t ed by the Georgia D e p a r t m e n t of Natm'al Resom'ces, Envfl 'omnental Pro- tection and Coastal Resom'ces Divisions.

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Toxaphene Residues in Terry and Dupree Creeks 595

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Received for conside.ration, t @ 19, 2000 Accepted for p~tblication, March 6, 2001