Science of the Total Environment 407 (2009) 3899–3904

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Perfluorinated contaminants in fur seal pups and penguin eggs fromSouth Shetland, Antarctica

A. Schiavone a,⁎, S. Corsolini a, K. Kannan b, L. Tao b, W. Trivelpiece c, D. Torres Jr. d, S. Focardi a

a Department of Environmental Science G. Sarfattiá, University of Siena, via P.A. Mattioli, 4, I-53100 Siena, Italyb Wadsworth Center, New York State Department of Health, and Department of Environmental Health Sciences, School of Public Health, State University of New York at Albany, Empire StatePlaza, P.O. Box 509, Albany, NY 12201-0509, USAc U.S. Antarctic Marine Living Resources Division, Southwest Fisheries Science Center, 8604 La Jolla Shores Drive, PO Box 271, La Jolla, CA 92037, USAd Instituto Antartico Chileno (INACH), Plaza Munoz Gamero, 1055, Punta Arenas, Chile

⁎ Corresponding author. Tel.: +39 0577 232 882; fax:E-mail address: schiavone4@unisi.it (A. Schiavone).

0048-9697/$ – see front matter © 2009 Elsevier B.V. Adoi:10.1016/j.scitotenv.2008.12.058

a b s t r a c t

a r t i c l e i n f o

Article history:

Perfluorinated compounds Received 18 November 2008Received in revised form 19 December 2008Accepted 19 December 2008Available online 24 March 2009

Keyword:Perfluorinated compoundsPenguinEggSealAntarctica

(PFCs) have emerged as a new class of global environmental pollutants. In thisstudy, the presence of perfluorochemicals (PFCs) in penguin eggs and Antarctic fur seals was reported for thefirst time. Tissue samples from Antarctic fur seal pups and penguin eggs were collected during the 2003/04breeding season. Ten PFC contaminants were determined in seal and penguin samples. The PFCconcentrations in seal liver were in the decreasing order, PFOSNPFNANPFHpANPFUnDA while in Adéliepenguin eggs were PFHpANPFUnDANPFDANPFDoDA, and in Gentoo penguin eggs were PFUn-DANPFOSNPFDoDANPFHpA. The PFC concentrations differed significantly between seals and penguins(pb0.005) and a species-specific difference was found between the two species of penguins (pb0.005). Inour study we found a mean concentration of PFOS in seal muscle and liver samples of 1.3 ng/g and 9.4 ng/gwet wt, respectively, and a mean concentration in Gentoo and Adélie penguin eggs of 0.3 ng/g and 0.38 ng/gwet wt, respectively. PFCs detected in penguin eggs and seal pups suggested oviparous and viviparoustransfer of PFOS to eggs and off-springs.

© 2009 Elsevier B.V. All rights reserved.

1. Introduction

Perfluorinated compounds (PFCs) have emerged as a new class ofglobal environmental pollutants. Several studies have reported theiroccurrence in wildlife (Giesy and Kannan, 2001; Olivero-Verbel et al.,2006; Van de Vijven et al., 2005), from low latitude to remote areas,suggesting their global distribution including open ocean waters andbiota (Prevedouros et al., 2006; Tao et al., 2006; Yamashita et al., 2008).

PFCs are persistent and bioaccumulative, although their physico-chemical properties are different from other known persistentorganohalogens. PFCs such as perfluorooctane sulfonate (PFOS) andperfluorooctanoic acid (PFOA), unlike organochlorines, do notaccumulate in lipids but concentrate in blood and liver tissues(Giesy and Kannan, 2001, 2002; Kannan et al., 2001a). PFOA andPFOS are peroxisome proliferators and elicit potent immunomodulat-ing effects in mice, involving thymic and splenic atrophy, loss ofthymocytes and splenocytes, and potent suppression of adaptiveimmune responses (Yang et al., 2002; Ishibashi et al., 2008a). Theexposure of rats to PFOA resulted in the suppression of genes involvedin inflammation and immunity (Guruge et al., 2006).

+39 0577 232 806.

ll rights reserved.

Marine mammals are sensitive to accumulation by persistent andbioaccumulative contaminants, because of their high trophic positionin the marine food web. Similarly, avian eggs have been used asindicators of environmental contamination. Much of the informationon the contamination by PFCs is from biological samples collected inNorthern Hemisphere (Kannan et al., 2001a; Van de Vijver et al., 2005;Ishibashi et al., 2008b); little is known about the current status andtemporal trend of PFC contamination in Antarctica (Giesy and Kannan,2001; Kannan et al., 2001b; Tao et al., 2006). In this study, we usedtissues of Antarctic fur seals and penguins to determine theconcentrations of PFCs in the Antarctic ecosystem. Antarctic fur sealsand penguins feed at the top of the polar marine food chain. Inaddition, they are non-migratory and non-nomadic species breedingin the Antarctic region; the tissue concentrations of chemicals are anindication of local contamination (Hollamby et al., 2006). Tissuesamples fromAntarctic fur seal pupswere collected from the carcassesfound dead during a season (2004) of high neonatal seal mortality.Penguin eggs samples were take from unhatched eggs, followingpermission from the Scientific Committee for Antarctic Research(SCAR) (Ahmed, 2003). The aim of this study was to determine theconcentrations of PFCs in Antarctic organisms. The detection of “newcontaminants” in remote regions such as Antarctica suggests theirwidespread distributions and highlighting the need to understandtransportation pathways and sources.

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2. Materials and methods

Twenty muscle, seventeen liver and five blubber samples werecollected from Antarctic fur seal (Arctocephalus gazella) pups betweenJanuary and February 2004, in the framework of the Italian Program ofResearch in Antarctica (PNRA) and of a National Science Foundation(NSF) expedition. Samples were collected on Livingston Island, SouthShetland, Antarctic Peninsula, (62°39′ S, 60°30′ W). Antarctic fur sealpups were found dead and the tissue samples were taken from thecarcasses at the time of necropsy. Samples had no signs ofdecomposition. Sampling location, gender, body weight, and bodylength were recorded (Table 1). Pups were aged based on their pelagestage (Kovacs and Lavigne, 1986). Unhatched eggs of Adélie penguins(Pygoscelis adéliae, n=13) and Gentoo penguins (Pygoscelis papua,n=13) were collected on King George Island, South Shetland,Antarctic Peninsula (62°10′ S, 58°67′ W), during the 2004/05 fieldseason. All samples were wrapped in polyethylene bags and stored at−20 °C until analysis.

2.1. Analytical methods and instrumental analysis for PFCs

PFCs were analyzed following the method described elsewhere(Kannan et al., 2001a; Tao et al., 2006), with somemodifications. PFCswere extracted by ion-pairing liquid extraction method. For eggsamples, 0.5 g of whole egg homogenate, 3 mL of Milli-Q water, 4 ng ofeach internal standard (PFBS, 13C4-PFOS, and 13C4-PFOA), 2 mL of0.25 M sodium carbonate buffer, and 1 mL of 0.5 M tetrabutylammo-nium hydrogen sulfate solution (adjusted to pH 10) were mixed in a15 mL polypropylene (PP) tube. The sample was then extracted with5 mL of methyl-tert-butyl ether (MTBE) by vigorous shaking for45 min. The MTBE layer was separated by centrifugation at 3500 rpmfor 5 min and then transferred into another PP tube (~4.5 mL). Theaqueousmixturewas rinsedwithMTBE and separated twice; all rinseswere combined in the second polypropylene tube. The MTBE extractwas evaporated to near-dryness under a gentle stream of nitrogen andthen reconstituted with 1 mL of methanol. For the extraction of liverand muscle samples, a small amount of tissue (about 1 g) washomogenized with 5 mL of Milli-Q water, and then 1 g of thehomogenate was transferred into a PP tube and extracted followingthe procedure described above. Matrix matched calibration standards(seven points ranging from0.5 to 75 ng/mL)were prepared by spikingdifferent amounts of calibration standards into sample matrix thatcontained no quantifiable amount of the target analytes; these

Table 1Details of fur seal pups found dead and used for collecting the samples.

Sample Date of sampling Tissue Age Sex Length (cm) Weight (Kg)

08 January 2004 M Steel born ♀ 68 4.609 January 2004 M, L 2 week old ♂ 66.5 410 January 2004 M 2 week old ♀ 65 4.311 January 2004 M 2 week old ♀ 71 4.212 January 2004 M, L 1 month old ♂ 77.5 613 January 2004 M, L 1 month old ♂ 73 4.614 January 2004 M, L 1 month old ♂ 74 4.615 January 2004 M, L 1 month old ♀ 67 3.616 January 2004 M, L Steel born ♂ 74 5.717 January 2004 M, L 45 days old ♂ 77 5.4518 January 2004 M, L 45 days old ♂ 77 619 January 2004 M, L 1 week old ♀ 65.5 420 January 2004 M, L 45 days old ♂ 74 5.321 January 2004 M, L 45 days old ♀ 77 5.322 January 2004 M, L 1 month old ♀ 71 4.323 January 2004 M, L 45 days old ♀ 65 3.6524 January 2004 M, L 2 months old ♂ 77 5.525 February 2004 M, L 2 months old ♀ 77 4.826 February 2004 M, L 2 months old ♀ 74 5.627 February 2004 M, L 2 months old ♀ 80 7.2

(M=muscle, L=liver).

standards were passed through the entire analytical procedurealong with the samples.

Analytes were detected and quantified using an Agilent 1100 serieshigh-performance liquid chromatograph (HPLC) coupled with anApplied Biosystems API 2000 electrospray triple-quadrupole massspectrometer (ESI-MS/MS). The MS/MS was operated in electrospraynegative ion mode. Target compounds were determined by multiplereaction monitoring (MRM). The MRM transitions were 299N80 forPFBS, 399N80 for PFHS, 499N99 for PFOS, 503N99 for 13C4-PFOS,599N99 for PFDS, 498N78 for PFOSA, 363N319 PFHpA, 413N369 forPFOA, 417N372 for 13C4-PFOA, 463N419 for PFNA, 513N469 for PFDA,563N519 for PFUnDA, and 613N569 for PFDoDA. Samples wereinjected twice to monitor sulfonates and carboxylates separately andPFBS was monitored in both of the injections. A midpoint calibrationstandard was injected after every 10 samples to check for theinstrumental response and drift. Calibration standards were injecteddaily before and after the analysis. Reported concentrations were notcorrected for the recoveries. Blanks were analyzed by passing Milli-Qwater and reagent through the entire analytical procedure. Blankscontained trace levels of PFOA in penguin egg analyses and trace ofPFOA and PFOS in seals sample analyses. The concentrations reportedhere were subtracted from the mean value found in blanks. To checkthe recovery,13C4-PFOS and 13C4-PFOAwere used as internal standards.For penguin eggs and seal tissue samples, mean recoveries of were 61%and 89% for 13C4-PFOS, and 133% and 75% for 13C4-PFOA, respectively.

The limit of quantitation (LOQ)was determined based on the linearrange of the calibration curve prepared at a concentration range of 0.1to 20 ng/mL. The tissue samples were compared to this unextractedstandard calibration curve. Because of the variety of matrices analyzedand because of evolving analytical methods, the LOQwas variable. TheLOQ was determined as the lowest acceptable standard in thecalibration curve, deemed acceptable if it was within ±30% of thetheoretical value and the peak area of the standardwas at least twice asgreat as thematrix blanks. The LOQwas 0.4 ng/g for all analytes in sealsamples, and 0.1 ng/g for all analytes in penguin eggs, except for PFOAand PFHpA, for which the LOQ was 0.2 and 0.5 ng/g respectively.

2.2. Statistical analysis

Statistical analyseswere performedwith STATISTICA 7 forWindows(Ver 7.1; Statsoft, Italia srl) at a significance level of p=0.05. Statis-tically significant differences between the mean concentrations ofcontaminants were investigated by a single factor one-way analysis ofvariance (ANOVA) and when significant differences were found, theywere tested among each other using Tukey's post hoc test. The absoluteconcentrationswere log10-transformedprior to statistical analysis if thedata did not follow a normal distribution (Levene test pb0.05). Thetransformed data were normally distributed and the Tukey test wasused to analyze the differences in concentrations between species,category of species or tissues. The overall significance level of eachanalysis was set at pb0.05. The Tukey–Kramer (HSD) test is one of anumber of post-hoc methods recommended for testing differencesbetween pairs of means among groups that contain unequal samplesizes (Zar,1999). The correlations between PFC concentrations and sealage groups were assessed by simple correlation analyses (Zar, 1999).Correlations are expressed using the Pearson correlation coefficient r. Ifa concentration of PFC was below the LOQ in a sample, the LOQ valueitself or a value of half the respective detection limit, was consideredprior to statistical analyses. The statistical analysis between seal tissueswere limited by the different number of samples (liver samples n=17,and muscle samples n=20).

3. Results and discussion

Fur seal samples were analyzed for the presence of fourperfluorinated sulfonic acids (PFSAs), and six perfluorinated

Table 2Concentrations (ng/g ww; mean±SD) of perfluorinated compounds in Antarctic samples (nd=not detected).

Fur seal pup, muscle Fur seal pup, liver Gentoo penguin egg Adélie penguin egg

% detected Mean±SD % detected Mean±SD % detected Mean±SD % detected Mean±SD

PFHS 0 nd 82 b0.4 0 nd 0 ndPFHpA 80 0.5±0.3 100 1.0±1.9 15 b0.5 54 2.5±5.5PFOS 100 1.3±0.7 100 9.4±3.2 100 0.3±0.1 100 0.4±0.2PFOSA 65 b0.4 100 b0.4 0 nd 54 0.2±0.3PFOA 50 0.8±0.8 6 b0.4 8 b0.2 23 b0.2PFNA 0 nd 94 3.3±1.7 0 nd 8 b0.1PFDA 0 nd 71 0.6±0.5 31 0.1±0.2 85 1.3±2.9PFDS 0 nd 6 b0.4 Not analyzed Not analyzedPFUnDA 0 nd 71 0.9±0.9 100 0.6±0.5 100 2.3±6.5PFDoDA 0 nd 6 b0.4 54 0.3±0.8 85 0.5±0.4PFCs 2.7±1 16±6.2 1.3±1.2 7.35±9

Fig. 1. ∑PFSA and ∑PFCA concentrations (ng/g ww) in seal and penguin samples.

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carboxylic acids (PFCAs); perfluorooctane sulfonate (PFOS), perfluor-hexanesulfonate (PFHS), perfluorooctanesulfonamide (PFOSA), per-fluorodecanesulfonate (PFDS), perfluorooctanoic acid (PFOA),perfluoroheptanoic acid (PFHpA), perfluorononanoic acid (PFNA),perfluorodecanoic acid (PFDA), perfluorododecanoic acid (PFDoA) andperfluoroundecanoic acid (PFUnDA). Penguin samples were analyzedfor all of these contaminants except for PFDS (Table 2).

3.1. PFC concentrations in seals: tissue distribution

Concentration of PFCs were significantly higher in liver than inmuscle (ANOVA pb0.001), suggesting compound-specific persistenceand retention of PFCs in seal liver. Mean concentration of ∑PFSAs(1.4 ng/g ww) was similar to the mean concentration of ∑PFCAs(1.3 ng/g ww) in seal muscle samples. Mean concentration of∑PFSAs (10.4 ng/g ww) in seal liver samples was 68% higher thanthat of∑PFCAs (6.2 ng/g ww), suggesting preferential enrichment ofPFOS in seal liver. Hepatic concentrations of PFOS were not correlated(pN0.05) with age.

The PFC concentrations in liver and muscle of seal samples were inthe decreasing order, PFOSNPFNANPFHpANPFUnDA and PFOSNP-FOANPFHpANPFOSA, respectively. A similar trend was previouslyreported in marine mammals (Martin et al., 2004; Van de Vijver et al.,2003), including polar bears (Smithwick et al., 2005; Kannan et al.,2005) from North America, where concentrations decreased withincreasing chain length, and the predominant PFC was PFOS. Seal liversamples contained significantly (pb0.001) higher concentrations ofPFOS than the muscle samples, while PFNAwas no detected in musclesamples. High PFOS concentrations (range b0.08–3.52 ng/mL ww)were also reported in blood samples of elephant seal pups from theSouth Shetland Islands (Tao et al., 2006). Previous studies havereported higher PFOS concentrations in dolphins and harbourporpoise pups than in adults (Houde et al., 2005; Van de Vijveret al., 2004; Hart et al., 2008). Some studies have clearly demonstrateda significant placental transfer of PFOS from dam to fetus duringgestation (in utero exposure to PFOS) and to the pup during lactation(Luebker et al., 2005a,b; Hart et al., 2008).

PFOA was detected at low concentrations (mean 0.8 ng/g ww) in50% of seal muscle samples, and between 6 and 23% in the othersamples analyzed. In general, PFOA has been detected sporadically inmarinemammals (Kannan et al., 2002b) and birds (Bossi et al., 2005a;Kannan et al., 2002c; Holmström and Berger, 2008).

3.2. PFCs in penguins: long-chain PFCA

Concentrations of PFCs were found in the decreasing orderPFHpANPFUnDANPFDANPFDoDA in Adélie penguin eggs, and PFUn-DANPFOSNPFDoDANPFHpA inGentoopenguin eggs. Several long-chainPFCAs (PFDA, PFUnDA, PFDoDA)were thepredominant PFCs detected inpenguin egg samples at the concentrations higher or no less than that of

PFOS. Thus, the mean concentrations of ∑PFCAs in Gentoo penguin(1.08ng/gww)andAdélie penguin eggs (6.6ng/gww)were anorderofmagnitude higher than ∑PFSAs (0.29 ng/g ww and 0.56 ng/g ww,respectively) (Fig. 1). PFCA profile was dominated by PFUnDA (mean0.59ng/gww) inGentoopenguin eggs, and by PFUnDA (2.33 ng/gww),and PFHpA (mean 2.53 ng/g ww) in Adélie penguin eggs, in which thePFCA concentrations were also higher than that of PFOS (mean 0.29,0.38 ng/gww, respectively). High concentrations of long-chain PFCAs ineggs suggest oviparous transfer of these compounds.

3.3. Correlations between PFCs

The statistical associations betweenPFOSA andPFOShave previouslybeen used to demonstrate that PFOSA is a precursor of PFOS (Kannanet al., 2002a; Martin et al., 2004). We found no statistically significantcorrelation between PFOS and PFOSA in Adélie penguin eggs (pN0.05).Other reports (Kannan et al., 2001b; Martin et al., 2004; Bossi et al.,2005) have found statistical association between PFOSA and PFOS fordifferent species, but not always consistent. It is not clear whether thisassociation represents metabolism or direct exposure. Statisticallysignificant linear correlations were only found between the concentra-tions of PFDA and PFUnA (pb0.05) in the seal liver (Fig. 2), suggesting acommon source of these two PFCAs to fur seals. A significant positiverelationship between concentrations of PFOS and PFNAwas observed infur seal liver (Fig. 3). These results indicate that the sources of exposureof the seal to PFOS and PFNA are similar, or the coexistence of thesecompounds in the sources.

3.4. Species-specific differences

The PFC concentrations differed significantly between seals andpenguins (pb0.005) and a species-specific difference was found

Fig. 2. Linear regression of PFDA and PFUnDA concentrations in fur seal liver fromAntarctica. A log transformation was performed to normalize the data.

Fig. 4. Concentrations of perflurooctane sulfonate (ng/g ww) in seal and penguinsamples from Antarctica. The straight line is the mean. The dots represent the outlier(N1.5 interquartile lengths from box edge). Statistically significant difference (pb0.005)is indicated by ⁎.

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between the two species of penguins (pb0.005) (Fig. 4). Although theseorganisms share the same environment, differences in diet andmetabolism might explain the variations in the concentrationsobserved. Reid and Arnould (1996) investigated the diet of lactatingfemale Antarctic fur seals (Arctocephalus gazella) in South Georgia,and found that Antarctic krill (Euphausia superba) was the main preyitem, followed by fishes and squid. The myctophids Protomyctophumchoriodon were the major diet during the lactational period. Outsidethis period, the nototheniid Lepidonotothen larseni and the chan-nichthyid Champsocephalus gunnari were the dominant prey species(Reid and Arnould, 1996). In contrast, differences in the PFC con-centrations between penguin species may be related to diet, repro-ductive status, ecological niches, and migration. Both penguin speciesin our study were from the South Shetland Islands, and feed primarilyon Euphausia superba, although Gentoo penguins feed more fish(Antarctic silverfish Pleuragramma antarcticum) than Adélie penguins.E. crystallorophias and pelagic and benthic species of amphipods areminor components of the pygoscelid diet (Volkman et al., 1980).Gentoo penguins feed inshore and are deep divers, while Adéliepenguins are shallow-diving, offshore foragers (Trivelpiece et al., 1987).

Among PFCs, PFOS was detected in all of the samples. PFOSconcentrations in the liver and muscle of seal pups were significantlyhigher (pb0.001) than those found in penguin eggs. No significantdifference was found in PFOS concentrations between eggs from the

Fig. 3. Relationship between concentrations of PFOS and PFNA in fur seal liver fromAntarctica.

two species of penguins (Fig. 5). PFOSA was below the detection limitin seal muscle and liver samples, and in Gentoo penguin eggs, but wasdetected in Adélie penguin eggs with a frequency of 54%. The PFCAprofile was dominated by PFUnDA in penguin eggs, in which the PFCAconcentrationswere also higher than that of PFOS. Similar results havebeen reported in other studies on birds (Holmström and Berger, 2008;Verreault et al., 2007).

3.5. Comparison with other studies

The presence of PFCs in penguin eggs and Antarctic fur seals isreported here for the first time. In fact, little information has beenprovided so far on PFC concentrations in these Antarctic organisms(Table 3). One previous study reported PFOS values under thedetection limit in Adélie penguin eggs (b0.1 ng/g ww; Tao et al.,2006), while in our study we found amean concentration of 0.38 ng/gww. This difference could be due to the different collection period,1995/1996 and 2004 respectively, suggesting an increase in theenvironmental levels of these contaminants. Tao et al. (2006) reporteda mean PFOS concentration in the blood of elephant seal pups of0.53 ng/mL— one order of magnitude lower than that found in our furseal pup liver samples (mean: 9 ng/g ww). This difference may due tothe preferential accumulation of PFCs in liver tissues (Van de Vijveret al., 2005).

Fig. 5. Log concentrations of PFOS (ng/g ww) in seal and penguin samples fromAntarctica. The straight line is the mean. The dots represent the outlier (N1.5interquartile lengths from box edge) Statistically significant difference (pb0.001) isindicated by ⁎.

Table 3Mean (Range) concentration of PFOS (ng/mL in blood or ng/g in other tissues ww) in seals and birds from Antarctica.

Matrix Date of collection Species n Location PFOS References

ww

Blood 2001 Pygoscelis adéliae 8 Admiral Bay b0.1 a Tao et al. (2006)(Adélie penguin) South Shetlands

2003/4 and 2004/2005 Mirounga leonine 59 Elephant Island 0.53 (b0.08–3.52) Tao et al. (2006)(Elephant seal) South Shetlands

Not available Stercorarius maccormicki 2 Terra Nova Bay 1.2 (b1a–1.4) Giesy and Kannan (2001)(Polar skua)

Egg 1995/1996 Pygoscelis adéliae 6 Edmonson point b0.1a Tao et al. (2006)(Adélie penguin)

1998/1999 Stercorarius maccormicki 3 2.51 (2.08–3.12)Polar skua

2004/05 Pygoscelis adéliae 17 Admiral Bay,King George Is.

0.38 (0.18–0.89) This study(Adélie penguin)Pygoscelis papua 20 0.28 (0.13–0.49)(Gentoo penguin)

Liver 2004/05 Arctocephalus gazella 17 Livingston Island 9.01 (1.85–17.25) This study(Antarctic fur seal)

Not available Leptonychotes weddellii 1 Terra Nova Bay b35 a Kannan et al. (2001b)(Weddell seal)

Muscle 2004/05 Arctocephalus gazella 20 Livingston Island 1.29 (0.42–3.59) This study(Antarctic fur seal)

a Concentration of PFOS were below LOQ for all samples.

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In comparison to seals and birds from the northern hemisphere,the concentrations of PFOS and PFOA in penguins and seals from theSouth Ocean were 10–100 fold lower. Despite the species differences,concentrations of PFOS measured in the liver of fur seals in our studywere two orders of magnitude lower than concentrations in liver ofharbor seals (162 ng/g ww) from the northwest Atlantic (Shaw et al.,2006) and one order of magnitude lower than that found in ringedseals (67 ng/g ww) from Greenland (Bossi et al., 2005). The PFOSconcentrations found in our penguin eggs were two orders ofmagnitude lower than the concentrations in herring gull eggs(52 ng/g ww) from northern Norway (Verreault et al., 2007), andthree orders of magnitude lower than that found in glaucous gull eggs(104 ng/g ww) from Svalbard (Verreault et al., 2005) and in guillemoteggs (614 ng/g ww) from the Baltic Sea (Holmström et al., 2005).However, comparisons should be interpreted with caution due tointerspecies differences in feeding ecology.

Although the contamination levels of PFOS and PFOA are low insouthern hemisphere fauna, the occurrence of PFCs in these remotelocations suggests the widespread distribution of PFCs. Antarcticorganisms should continue to be monitored in future due to the globaltrend of increasing PFC use.

3.6. Ecological implications

Acute and chronic dietary exposure studies of PFOS in mallardducks (Anas platyrhynchos) and northern bobwhite quails (Colinusvirginianus) (Newsted et al., 2007, 2006) have recently led to thecalculation of PFOS toxicity reference values (TRVs) and predicted no-effect concentrations (PNECs), based on the characteristics of topavian predators (e.g. birds of prey and certain gull species) (Newstedet al., 2005). Conservative egg yolk-based TRVs and PNECs weredetermined as 1.7 and 1.0 μg PFOS/mL, respectively. In the same study,the lowest observable adverse effect level (LOAEL) in egg yolk was62.0 μg PFOS/mL. Hence, the mean PFOS concentration evaluated inpenguin eggs from Antarctic Peninsulawas approximately 3000 timeslower than the PFOS TRV, PNEC, and LOAEL values, respectively. From atoxicological standpoint, and assuming that the sensitivity to PFOSexposure of mallard ducks and northern bobwhite quails is similar inpenguins, recent concentrations in eggs suggest that PFOS alonewould pose a minimal risk to the developing penguin embryo.However, PFOS and other accumulated PFASs in penguins need to beassessed as a part of a broad contaminant cocktail, including chlorine-

and bromine-based chemicals with potential health risks (Newstedet al., 2007, 2006; Molina et al., 2006).

Other laboratory studies on rats, monkeys and birds have shownthat the toxic effects of PFOA and PFOS occur at tissue concentrationsin the range of a few tens to hundreds of μg/g ww (Kennedy et al.,2004; Newsted et al., 2006). Residue concentrations of PFOS and PFOAin our fur seal livers were approximately 3–4 orders of magnitudelower than the effect concentrations found in laboratory animals. Thetoxic effects of PFCs in seal species are unknown, as only one study hasfound a significant association of PFOS and PFOA with the presence ofdisease mortality in sea otters (Kannan et al., 2006): establishment ofa cause-effect linkage will require toxicological and controlled animalfeeding studies. Further studies are needed on the immunotoxiceffects of PFCs and also on the interaction between PFCs and othercontaminants found in fur seal tissues.

Acknowledgments

This research was funded by the Italian National Program ofResearch in Antarctica (PNRA). The National Science Foundationsupported S. Corsolini's stay and travel to and from King George Is. Weare very grateful to Daniel Torres and Daniel Torres jr (InstitutoAntarctico Chileno, Santiago, Chile) for collecting the fur seal samplesduring the 2003/04 expedition, and to Wayne Trivelpiece, and SusanTrivelpiece for collecting the penguin eggs samples. We thanks RogerHewitt, the Agunsa (Punta Arenas, Chile) and Raytheon (USA) fortheir friendly logistic support.

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