Polar Biol ,2002) 25: 206±215 DOI 10.1007/s00300-001-0328-9

ORIGINAL PAPER

S. Duquesne á M.J. Riddle Biological monitoring of heavy-metal contamination in coastal waters off Casey Station, Windmill Islands, East Antarctica

Accepted: 17 September 2001 / Published online: 14November 2001 Ó Springer-Verlag 2001

Abstract Heavy-metal concentrations were determined formation gathered about processes of contaminant in tissues of di€erent species of benthic invertebrates uptake and partitioning among di€erent tissues and collected in the Casey region ,Australian Antarctic species could be used in later studies to investigate the Territory) where an old waste-disposal tip site is a source behaviour and the source of contaminants. of contamination. The species studied included the biv- alve Laternula elliptica, star®sh Notasterias armata, heart urchins Abatus nimrodi and A. ingens and gam- Introduction maridean amphipod Paramoera walkeri. The specimens were collected at both reference and contaminated lo- Antarctica is the most remote and least-inhabited con- cations where lead was the priority element and copper tinent; however, it is no longer free of environmental was the next most important in terms of increased contaminants. Processes such as ``global distillation'' concentrations. The strong association between a gra- cause the transfer of pollutants, particularly volatile dient of contamination and concentrations in all species hydrocarbons, from the industrialized regions of the tested indicates that they are re¯ecting well the envi- world to the polar regions ,Risebrough et al. 1976; Van ronmental changes, and that they appear as appropriate den Brink 1997). More locally, activities in the Antarc- biological indicators of heavy-metal contamination. tic, such as shipping activities and research stations, Aspects of the biology of species with di€erent func- have created sites with contamination at levels compa- tional roles in the marine ecosystem are discussed in rable to industrial sites elsewhere ,Berkmann 1992; relation to their suitability for wider use in Antarctic Deprez et al. 1999). In response to these, all countries monitoring programmes. For example, in terms of operating in Antarctica have now agreed to a range of heavy-metal bioaccumulation, the bivalve appears as the measures to reduce or eliminate pollution ,Madrid most sensitive species to detect contamination; the protocol), and this requires the monitoring of levels of star®sh provides information on the transfer of metals contaminants. through the food web while the heart urchin and The present investigation deals with a contaminated gammarid give indications of the spatial and temporal site at a research station. In this case-study, there is patterns of the environmental contamination. The in- evidence showing that trace metals are the contaminants of most concern in the soil of an old onshore waste- disposal site ,Deprez et al. 1999). They are mobilized by meltwater during the summer thaw and ¯ow in to the S. Duquesne ,&) National Research Centre for Environmental Toxicology, adjacent bay, partly dissolved but mostly adsorbed to University of Queensland, Kessels Road, particles on land and in the marine environment ,Snape Coopers Plain, QLD 4108 Australia et al., in press). Analyses in leachate showed that lead, M.J. Riddle copper and, to a lesser extent, zinc are of concern Australian Antarctic Division, Channel Highway, ,Deprez et al. 1999). Analyses in marine sediment Kingston, TAS 7050, Australia showed that lead and copper concentrations are about Present address: S. Duquesne 17- and 3.5-fold more elevated than background values University of the West of England, whereas no signi®cant increase is detected for zinc. They Faculty of Applied Sciences, Frenchay Campus, also demonstrate the presence of a gradient of contami- Coldharbour Lane, Bristol, BS16 1QY, England e-mail: [email protected] nation among some of the sites currently investigated Tel.: +44-117-3442680 ,S. Duquesne and M. Liess, unpublished data). Strong Fax: +44-117-9763871 evidence supports the fact that the increase of metal 207 concentrations in the nearshore environment is due to also collected at other locations further away from the tip site, the vicinity to the old waste-disposal tip site rather than i.e.Newcombe Bay ,about 1.5±3 km from the tip), Shannon Bay to a natural background enrichment of the area, because ,about 1.5 km) and O'Brien Bay ,about 4km) ,Fig. 1). ,1) waters became enriched in contaminants compared to background levels when they ¯owed through the tip Collection and dissection of animals down towards the sea ,Snape et al., in press), ,2) only 2 out of 16 sites investigated along the shore had signi®- Bivalves, star®sh and heart urchins were collected by divers at all cantly high amounts of metals in suspended particulates; locations, at depths between about 5 and 20 m. Then individuals of both sites were adjacent to old waste-disposal tip sites sizes from similar ranges were selected. Animals were brought back alive to the laboratory and kept in aquaria in clean ®ltered seawater ,Wilkes and Old Casey) ,Liess and Duquesne 1997), and at 0‹0.5°C for 2 days before dissection. Di€erent tissues were ,3) the chemical pro®les of contaminants at sea and in dissected from the various species ,gills, kidney and muscle-siphon the soil of the tip site are similar. from the bivalve; pyloric caeca and stomach from the star®sh; di- Monitoring the levels of contaminants using biologi- gestive tract and skeleton from the heart urchin). After dissection, all tissues were stored in polyethylene bags and frozen at ±20°C. cal species will provide a measure of the fraction of The size and sex of each individual were recorded. The mean sizes chemicals that is bioavailable and thus potentially toxic. were 79.2‹19.4 mm for bivalves, 40.5‹4.6 mm for heart urchins Such monitoring is widely used ,Phillips 1980; Stebbing and 30.25‹4.71 cm for star®sh. et al. 1992). Suitable benthic invertebrates for use Gammarids were collected at a reference site with hand nets and as biological indicators of trace-metal contamination in transplanted to two sites within Brown Bay ,Fig. 1, sites a and b). Adults ,1±1.5 cm) were placed in plastic enclosures ,15 l) with aquatic environments include bivalves ,Bryan et al. 1-mm-mesh netting at the top and bottom. These enclosures were 1985), echinoderms ,Warnau et al. 1995; Temara et al. held vertically at 1 m below the water surface and four of them 1998a) and crustaceans ,Rainbow and White 1989; were placed at each location. Each of them contained 50 individ- Moore et al. 1991; Zauke et al. 1995). The species selected uals. After 18 days of exposure, individuals from a single enclosure were kept in aquaria in clean ®ltered seawater at 0‹0.5°C for in this study were the bivalve, Laternula elliptica, two 2 days. They were then pooled, rinsed with distilled water and heart urchins ,Abatus nimrodi and A. ingens), the aster- frozen at ±20°C. oid, Notasterias armata, and the crustacean, Paramoera walkeri, as they are common in nearshore waters of the investigated region. Furthermore, some of them are Analyses of heavy metals known to be distributed around the Antarctic continent. All samples were dried at 50°C under vacuum and about 0.5 g of The use of these species will provide information on the dry material was digested for 12 h at room temperature in 3 ml the biological availability of metals to species with a 70% nitric acid ,Aristar) in polyethylene-capped tubes. Samples variety of habitats and feeding modes and may indicate were then heated at 90°C in a water bath for 3 h and ®nally diluted how metals are distributed among di€erent environ- to a total volume of 40 ml with distilled water. Blanks and standard mental compartments and accumulated in the environ- reference material ,NIST, oyster 1566a) were processed in the same way. Extra samples from each type of tissue were spiked with in- ment ,Rainbow 1985). ternal standards for calculation of metal concentrations. Prior to The objectives of this study were: ,1) to assess the digestion, a fraction of heart-urchin skeleton was ®rst cleansed of extent of contamination in a suite of representative non-calci®ed tissues using 1& ,w/v) proteinase N ,Serva) solution Antarctic nearshore invertebrates; this baseline study will according to the method of Dubois and Jangoux ,1985). Metal concentrations ,arsenic, cadmium, chromium, copper, be used for monitoring changes after clean-up operations, nickel, lead and zinc) were determined by ICP MS using a Perkin as part of remediation procedures of contaminated sites; Elmer Elan 5100 for all samples except the gammarids, which were and ,2) to determine whether the species tested are useful analysed on a Fissons PQ2+STE. Analyses of certi®ed reference as biological indicators of heavy-metal contamination, material ,SRM 1566a) were in the range of 87±112% of the certi®ed for further monitoring programmes in Antarctica. values, indicating that analyses were both accurate and precise.

Statistical analyses

Materials and methods Di€erences in metal concentrations of specimens from di€erent sites ,contaminated or control), tissues and species were tested Study area using analysis of variance ,ANOVA). The numbers of specimens collected at the di€erent locations varied ,between 2 and 16, see The region around Casey Station, Wilkes Land, East Antarctica Table 1); hence the numbers of replicates used in the analysis were ,110°30¢E,66°17¢S), a small ice-free site in an area of peninsulas selected to make best use of the available samples. Some sites were and rocky islands, was selected for this study because there are excluded from the analysis because too few replicates were avail- several discrete sites nearby representing the full range of con- able. At other sites, some specimens were excluded from the anal- tamination typical of Antarctic operations. ysis at random to ensure a balanced design, with equal numbers of This investigation was carried out during the summers of 1996/ replicates for each treatment. Prior to ANOVA, all data were tested 1997 and 1997/1998 in Brown Bay, a small embayment adjacent for homogeneity of variance using Cochrane's test. Where signi®- to the old waste-disposal tip from the Old Casey, a station now cant heterogeneity could not be corrected with a transformation, disused. This tip site contains wastes generated from domestic, the signi®cance of the associated F-test was reduced by one level. workshop and laboratory activities ,Deprez et al. 1999). Data from analysis of the bivalve L. elliptica and the star®sh Animals were collected at two sites in Brown Bay ,sites 1 and 2). N. armata were used in two-factors ANOVA to test for e€ects of Site 1 is within 30 m of the point where meltwater enters the bay, location and tissue type ,two locations and three tissue types for whereas site 2 is about 400 m from the tip outlet. Specimens were bivalve; four locations and two tissue types for star®sh). For the 208 combinations with one datum missing, the mean was substituted in animals from Brown Bay ,close to the source of for the missing datum and the degrees of freedom used in the F-test contamination) than from the other locations. reduced accordingly. Data from the heart urchins ,A. nimrodi and A. ingens) were In all species and tissue types from the Brown Bay used in two separate analyses. Data from the analysis of the di- sites, Pb was present at signi®cantly higher concentra- gestive tract of A. nimrodi were used in a one-factor ANOVA to tions than at the other locations. Cu and Zn concen- test for the e€ect of location using data from ®ve sites. Data from trations were also signi®cantly higher in specimens from both A. nimrodi and A. ingens were used in a three-factor ANOVA Brown Bay sites for most species and tissue types. The to test for the e€ects of location ,2), species ,2) and tissue type ,2). When ANOVA provided signi®cant results, the Student-New- only exceptions to this general pattern were that Cu was man-Keuls test was used as post-hoc test. not signi®cantly higher in N. armata from Brown Bay The homogeneity of sizes at the di€erent locations was tested and Zn was higher in the skeleton of heart urchins from using ANOVA, as well as the dependence between sex and metal the reference site ,O'Brien Bay). Most metal concen- concentrations. The dependence between size and metal concen- trations was evaluated using linear regressions and ANOVA. trations decreased as follows: Brown Bay>Shannon Data from the gammarid P. walkeri deployed in enclosures for and Newcombe Bay>O'Brien Bay. However, Ni and Cr 18 days were compared using a Student's t-test to test for the e€ect were not found at signi®cantly higher concentrations in of location ,Brown Bay sites a and b). individuals from Brown Bay. Also, arsenic in both tissue types of N. armata ,Table 1) and Cr and Ni in the di- gestive tract of A. nimrodi ,data not shown) were found Results at signi®cantly higher concentrations in individuals from reference locations ,O'Brien Bay). Di€erences in metal concentrations among locations In the gammarid, P. walkeri, Pb was the only metal in animal tissues present at signi®cantly higher concentrations in individ- uals kept in enclosures close to the tip outlet ,site a) than in Di€erences in metal concentrations among locations those kept further away ,site b) ,t-test, P<0.05) ,Table 2). were signi®cant for 21 of the 27 combinations of species ,bivalves, heart urchins and star®sh) and metals ,Table 1 Fig. 1 Antarctica and the Casey region with bays used for and data not shown). Sixteen of the 21 signi®cant dif- collection [two sites were used for collections in Brown Bay ,sites ferences were because metal concentrations were higher 1 and 2) and gammarids were transplanted to sites a and b] Table 1 Di€erences in concentrations of heavy metals in tissues from Laternula elliptica ,kidney Kid, gill Gill, muscle Mus), Notasterias armata ,pyloric caeca Pc, stomach Stom) and heart urchins ,Abatus ingens Ai, Abatus nimrodi An, digestive Dig, skeletal Sk tissues). Individuals were collected at sites adjacent to a source of contamination ,Brown Bay sites 1 and 2 BB1, BB2) and reference sites ,O'Brien Bay OB and Shannon Bay SH) and tested using analysis of variance and SNK test ,Student-Newman-Keuls test) ,ns not signi®cant)

Metal Location Tissue Location ´ Tissue Species Location ´ Species Species ´ Tissue Location ´ Species´Tissue

Bivalve Laternula elliptica Arsenic *BB1>SH ***Kid>Gill>Mus ns Cadmium ns ***Kid>Gill>Mus ns Chromiuma ns ns ns Coppera **BB1>SH **Kid>Gill>Mus **BB>SH ,Kid, Mus only) Kid>Gill=Mus ,BB1) Kid>Gill>Mus ,SH) Nickela ns ns ns Lead ***BB1>SH ***Kid>Gill>Mus ***Kid>Gill>Mus ,BB1) Kid>Mus=Gill ,SH) Zinc **BB1>SH ***Kid>Gill>Mus ns Star®sh Notasterias armata Arsenic ***OB>SH ***Pc>Stom **OB>SH=BB1=BB2 ,Pc) =BB1>BB2 OB>BB2;BB1>BB2 ,Stom) Cadmium ns **Pc>Stom ns Chromium ns ***Stom>Pc ns Copper ns ***Pc>Stom ns Lead ***BB1=BB2 ***Stom>Pc **BB1=BB2>OB=SH ,Pc) >SH=OB BB1=BB2>SH>OB ,Stom) Stom>Pc ,all except OB) Zinc *BB2>OB ns *BB2=BB1=SH>OB ,Stom) Pc>Stom ,OB) Heart urchins Abatus nimrodi and Abatus ingens Arsenic ***BB1>OB ***Dig>Sk ***BB1>OB ,Dig) ***Ai>An ns ***Ai>An ,OB ns only) Dig>Sk ,An, Ai) Cadmium *BB1>OB **Dig>Sk *BB1>OB ,Dig) ns ns ns ns Chromium **OB>BB1 **Dig>Sk **OB>BB1 ,Dig) ns *Ai>An ,BB1) ns *OB>BB1 ,An, Dig) An>Ai ,OB) OB>BB1 ,Ai, Dig) Ai>An ,BB1, Dig) An>Ai ,OB, Dig) Coppera **BB1>OB **Dig>Sk *BB1>OB ,Dig) ns *Ai>An ,BB1) ns *BB1>OB ,An, Dig) BB1>OB ,Ai, Dig) Ai>An ,BB1, Dig) Nickel **OB>BB1 ***Dig>Sk **OB>BB1 ,Dig) ns *Ai>An ,BB1) ns **OB>BB1 ,An, Dig) An>Ai ,OB) OB>BB1 ,Ai, Dig) Ai>An ,BB1, Dig) An>Ai ,OB, Dig) 209 210 Di€erences in metal concentrations among species

This comparison can only be applied to the heart ur- chins, as only in the two species of Abatus were exactly the same tissues used. The concentrations of all elements measured did not vary signi®cantly between the two species ,Table 1), except that As was present at signi- ®cantly higher concentrations in A. ingens than in A. nimrodi.

Di€erences in metal concentrations among tissue types

Tissue type was a highly signi®cant factor a€ecting the concentrations of most metals in all species ,Table 1). In the bivalve L. elliptica, As, Cd, Cu, Pb and Zn were consistently highest in the kidney and lowest in the muscle while Cr and Ni did not vary signi®cantly among the tissue types. In the star®sh N. armata, As, Cd and Cu were higher in the pyloric caeca while Cr and Pb were higher in the stomach. In both species of heart urchins, A. nimrodi and A. ingens, all elements were signi®cantly higher in the digestive tract than in the skeleton. The ANOVA indicated that there are some signi®- cant interaction e€ects of tissue with one or more of the other factors tested ,location, species); however, these did not contradict the overall patterns described above ,Table 1).

Metal concentrations in di€erent species and in¯uence of sex and size of individuals

The mean metal concentrations for each type of species, tissue and location are reported in Table 2. Statistical results showed that there was no dependence between

Dig>Sk ,BB1, OB) BB1>OB ,Dig) Dig>Sk ,BB1, OB) sex and metal concentrations ,data not presented). Star®sh and heart-urchin mean sizes did not di€er sig- ni®cantly among locations ,ANOVA, P>0.05) whereas bivalve sizes did ,ANOVA, P<0.05; 79.2‹10.1 mm in Brown Bay 1 and 103.2‹12.3 mm in Shannon Bay). Although the bivalves were larger at Shannon Bay, one can assume that this did not in¯uence the metal con- centrations, as there was no signi®cant correlation be- tween sizes and concentrations for individuals originated from 1 location ,for example, in Brown Bay 1: n=10; P>0.05; r2<0.18, 0.24and 0.14in gills, kidney and muscle, respectively, for all elements tested; data not presented). **BB1>OB**BB1>OB **Dig>Sk **Dig>Sk **BB1>OB ,Sk, Dig) **OB>BB1 ,Sk) ns ns ns ns ns ns ns ns

Relative increases of metal concentrations among locations, species and tissue types

The relative increases of Pb, Cu and Zn concentrations

a in animals collected at the site adjacent to the source of a <0.001 <0.01;

P contamination ,Brown Bay 1) compared to those from <0.05; P Lead Zinc P Heterogeneity of variance could not be fully corrected by transformation and therefore level of signi®cance is reduced by 1 reference sites ,Shannon and O'Brien Bays) are repre- * *** a ** 211 sented by a ratio ,Table 3). The elements with most concentrations of speci®c metals in specimens collected signi®cant increases at the contaminated site were Pb, close to the source of contamination indicates that the then Cu and then Zn. selected species are ecient accumulators of contami- The ratio in various tissues decreased as follows: nants, and thus satis®es a basic requirement to be ap- bivalve gills >heart-urchin digestive tract >bivalve propriate biological indicators ,Farrington et al. 1983). muscle and kidney >star®sh tissues for Pb ,BB1/SH); heart-urchin digestive tract >bivalve muscle and kidney >tissues of star®sh and bivalve gills for Cu ,BB1/SH); Relevance of the di€erent biological species heart-urchin digestive tract >heart-urchin skeleton as biomonitors of metal concentrations >star®sh tissues for Pb ,BB1/OB) and heart-urchin di- gestive tract >star®sh stomach >heart-urchin skeleton The lamellibranch bivalve, L. elliptica, is a ®lter-feeding and star®sh pyloric caeca for Cu ,BB1/OB). organism and has been suggested as a biological indi- For gammarids deployed at the two sites in Brown cator of contamination in the Antarctic nearshore en- Bay, only Pb accumulated signi®cantly in individuals vironment ,Ahn et al. 1996). It feeds by drawing a closer to the waste-disposal site ,about 3.8) ,Table 3). current of water through the siphon and ®ltering out particulates in gills. This mode of feeding predisposes the animal to uptake contaminants that are both dis- solved and adsorbed to suspended particles. Although Discussion the animal lives buried in the sediment, it is unlikely to be exposed to high levels or pulses of contamination General contamination and signi®cance once particles with adsorbed contaminants have settled to the seabed, except as a consequence of resuspension This study deals with marine invertebrates collected in an ,Ahn 1997). area adjacent to an old waste-disposal tip site. Previous Levels of As, Cu, Pb and Zn were all higher in studies support the evidence that this coastal area is tissues of L. elliptica collected close to the source of contaminated by heavy metals leaching from the tip, contamination ,Brown Bay) than in those located particularly Pb and Cu. Indeed these elements were further away ,Shannon Bay). There was a consistent detected in tip material and leachate ,Deprez et al. 1999) pattern of higher concentrations in kidneys than gills and were enriched in marine sediment and seawater than muscle. The gills usually respond rapidly to ,dissolved and particulate phases) from the adjacent bay variations and pulses of dissolved or particulate-bound ,Snape et al., in press). However, their concentrations are contaminants and accumulate them both actively un- not expected to be of risk for the biota. Indeed, E€ects der metabolic control ,absorbed) and passively Range-Median from Sediment Guidelines values ,Long through adsorption ,Rainbow 1985). Thus metal et al. 1995) are above measured concentrations in this concentrations may be lower in gills than in kidney location ,44.7 and 25.3 ppm for Pb and Cu, respectively, because of low contamination at the time of sampling at the contaminated site vs 2.6 and 6.9 ppm for Pb and Cu or because, as in many bivalve species, the kidney is at the reference site of O'Brien Bay; S. Duquesne and the target for metal accumulation ,Langston and M. Liess, unpublished data). Concentrations measured in Zhou 1987; Sullivan et al. 1988; Serra et al. 1995). seawater were about 4 lgl±1 for Pb and Cu ,Snape et al., in Indeed, the response of kidney is slower than that of press) and previous work on the gammarid P. walkeri gills but its levels can build up as it acts as temporary reported 10,000 lgPbl±1 as not highly toxic, and a 4-day or permanent storage sites for metals absorbed with LC50 of 970 lgCul±1 ,Duquesne et al. 2000). food or water ,Depledge and Rainbow 1990). Many Although the contamination may not be toxic to the species can tolerate high metal levels in tissues such as biota, the metals are expected to bioaccumulate. Indeed, kidney because of the ability to regulate intracellular at the time of the investigation, the contaminants re- concentrations using detoxi®cation mechanisms such leased from the source of contamination in the near- as the production of insoluble inorganic-rich granules, shore environment should be dispersed throughout the membrane-limited vesicles or metalloproteins ,Viar- water column as a result of clearance of sea ice and, engo 1985). Among the three tissues, the muscle is consequently, of the disappearance of water-column expected to respond to a lesser extent to changes of strati®cation. They should thus become available to environmental concentrations, and this is con®rmed by surface-living, planktonic and benthic species, such as our data. the gammarid, P. walkeri. Particles with adsorbed con- The two species of heart urchins, A. nimrodi and taminants that settle should become available to ®lter A. ingens, live beneath the surface of soft sediment and feeders such as the bivalve, L. elliptica, for a brief period are selective deposit feeders. They may be exposed to and then once on the seabed, to sediment-consuming sediment-bound contaminants both through consump- deposit feeders such as the heart urchin Abatus sp. tion and by contact with their body surface. Echinoids This study shows that metals present in the environ- do not osmoregulate eciently and have no excretory ment are being taken up by the biota and are, therefore, organs; they are thus likely to re¯ect changes of envi- bioavailable. The consistent pattern of elevated ronmental conditions. 212

Table 2 Concentrations of metals measured in tissues from Ant- Bay=Newcombe Bay >O'Brien Bay). Values for Paramoera arctic benthic invertebrates collected at sites along a putative gra- walkeri refer to concentrations in individuals transplanted for dient of contamination ,Brown Bay 1>Brown Bay 2>Shannon 18 days to 2 sites in Brown Bay ,a and b),Nd not determined)

Site n Concentrations ppm dry weight ,standard deviation)

Arsenic Cadmium Chromium Copper Nickel Lead Zinc

Bivalve Laternula elliptica Kidney Brown Bay 1 10 145.4 ,18.0) 38.9 ,6.7) 4.3 ,0.7) 51.1 ,13.5) 9.7 ,2.4) 19.4 ,2.7) 170.5 ,28.7) Kidney Shannon Bay 4 114.4 ,26.9) 33.5 ,11.4) 4.3 ,1.4) 23.7 ,3.5) 7.3 ,2.2) 2.8 ,0.4) 148.6 ,11.8) Gills Brown Bay 1 10 76.9 ,14.4) 12.2 ,5.8) 3.6 ,0.8) 11.3 ,6.1) 3.2 ,1.4) 7.1 ,3.5) 143.3 ,24.5) Gills Shannon Bay 467.2 ,27.6) 20.2 ,7.3) 3.5 ,1.5) 8.8 ,0.6) 3.5 ,1.3) 0.36 ,0.08) 129.3 ,0.9) Muscle Brown Bay 1 10 26.4,9.2) 9.9 ,2.9) 22.0 ,18.8) 7.8 ,1.9) 12.3 ,9.6) 4.6,5.2) 106.0 ,13.6) Muscle Shannon Bay 3 15.5 ,5.5) 4.9 ,3.7) 2.0 ,0.5) 3.1 ,0.6) 0.6 ,0.1) 0.46 ,0.21) 73.6 ,16.3) Heart urchin Abatus nimrodi Digestive tract Brown Bay 1 8 36.5 ,3.9) 1.2 ,0.3) 55.7 ,21.7) 17.1 ,3.2) 25.7 ,10.0) 46.9 ,6.9) 73.1 ,6.7) Digestive tract Brown Bay 2 16 38.6 ,5.2) 1.1 ,0.3) 93.7 ,52.6) 12.3 ,2.1) 43.3 ,25.3) 20.9 ,5.7) 63.3 ,3.6) Digestive tract Shannon Bay 9 36.2 ,4.3) 1.6 ,0.3) 124.1 ,43.0) 8.3 ,1.0) 57.6 ,20.8) 4.7 ,0.9) 60.6 ,4.4) Digestive tract Newcombe Bay 7 29.1 ,4.7) 1.4 ,0.2) 123.1 ,71.0) 7.1 ,1.0) 44.9 ,9.6) 1.2 ,0.3) 58.1 ,3.5) Digestive tract O'Brien Bay 5 14.2 ,3.0) 0.69 ,0.14) 267.6 ,53.8) 9.8 ,1.5) 132.8 ,31.6) 1.0 ,0.0) 27.5 ,4.6) Skeleton Brown Bay 1 8 0.07 ,0.03) 0.10 ,0.01) 0.33 ,0.08) 2.5 ,1.2) 0.66 ,0.10) 1.40 ,0.57) 3.6 ,0.7) Skeleton Brown Bay 2 16 0.08 ,0.02) 0.09 ,0.01) 0.24,0.06) 2.4,1.0) 0.67 ,0.24) 1.22 ,0.53) 3.6 ,1.0) Skeleton Newcombe Bay 5 0.10 ,0.02) 0.09 ,0.01) 0.13 ,0.02) 4.0 ,1.1) 0.68 ,0.04) 0.05 ,0.04) 4.8 ,0.8) Skeleton O'Brien Bay 6 0.04 ,0.02) 0.08 ,0.01) 0.12 ,0.02) 2.8 ,1.4) 0.52 ,0.04) 0.17 ,0.11) 3.3 ,0.4) Heart urchin Abatus ingens Digestive tract Brown Bay 1 5 73.2 ,21.5) 1.3 ,0.2) 97.3 ,50.3) 26.4 ,5.9) 43.3 ,22.0) 47.3 ,5.3) 83.8 ,12.1) Digestive tract Shannon Bay 6 67.6 ,17.3) 1.7 ,0.2) 117.9 ,42.7) 10.4 ,1.1) 54.9 ,20.3) 3.2 ,0.5) 55.4 ,5.4) Digestive tract Newcombe Bay 3 69.3 ,29.6) 2.2 ,0.2) 85.1 ,25.1) 7.0 ,0.5) 39.8 ,12.0) 1.4,0.4) 56.6 ,2.5) Digestive tract O'Brien Bay 11 21.9 ,6.6) 1.0 ,0.4) 171.5 ,43.4) 8.5 ,2.0) 82.8 ,21.8) 1.1 ,0.2) 34.8 ,11.8) Skeleton Brown Bay 1 5 0.22 ,0.19) 0.11 ,0.01) 0.35 ,0.06) 4.0 ,3.6) 0.71 ,0.11) 1.32 ,0.25) 2.9 ,0.4) Skeleton O'Brien Bay 12 0.09 ,0.05) 0.10 ,0.01) 0.13 ,0.03) 2.6 ,0.9) 0.64,0.12) 0.18 ,0.08) 3.5 ,1.0) Star®sh Notasterias armata Pyloric caeca Brown Bay 1 5 23.3 ,4.1) 2.5 ,0.7) 1.4 ,0.3) 32.7 ,14.3) Nd 1.1 ,0.2) 92.1 ,10.8) Pyloric caeca Brown Bay 2 5 22.6 ,4.8) 2.7 ,1.1) 1.5 ,0.2) 37.0 ,10.8) Nd 0.90 ,0.35) 103.5 ,9.3) Pyloric caeca Shannon Bay 5 27.4,5.2) 2.6 ,0.8) 1.4,0.3) 31.8 ,11.1) Nd 0.23 ,0.04) 89.1 ,13.9) Pyloric caeca Newcombe Bay 2 24.0 ,5.6) 1.3 ,0.6) 1.4 ,0.4) 28.3 ,16.2) Nd 0.54 ,0.29) 93.9 ,25.4) Pyloric caeca O'Brien Bay 7 39.5 ,6.9) 2.6 ,1.3) 1.5 ,0.2) 30.0 ,9.4) Nd 0.32 ,0.10) 105.7 ,35.6) Stomach Brown Bay 1 419.2 ,0.4)1.5 ,1.0) 2.5 ,0.6) 10.2 ,2.8) Nd 2.7 ,1.0) 103.7 ,5.2) Stomach Brown Bay 2 3 12.3 ,2.1) 1.9 ,1.0) 2.4,0.4) 7.0 ,0.7) Nd 2.5 ,0.7) 105.6 ,13.9) Stomach Shannon Bay 5 15.9 ,3.9) 3.0 ,0.9) 2.4,0.9) 7.0 ,1.4) Nd 0.73 ,0.18) 101.1 ,10.6) Stomach Newcombe Bay 2 14.1 ,0.3) 1.7 ,1.3) 2.7 ,0.6) 6.0 ,2.0) Nd 1.8 ,2.1) 104.0 ,24.9) Stomach O'Brien Bay 6 21.2 ,3.4) 1.6 ,0.5) 2.0 ,0.7) 6.2 ,1.4) Nd 0.44 ,0.17) 81.4 ,10.0) Gammarid Paramoera walkeri Whole body Brown Bay site a 4 Nd 4.94 ,0.30) Nd 26.3 ,2.06) 5.26 ,0.21) 3.39 ,0.89) 68.5 ,4.69) Whole body Brown Bay site b 2 Nd 5.11 ,0.06) Nd 26.3 ,0.59) 6.0 ,0.60) 0.90 ,0.04) 74.5 ,5.05)

The habitat and feeding modes of the two species are mechanism for uptake of contaminants is via food, top almost identical and so are their metal concentrations. predators will respond more slowly than ®lter feeders or Digestive tissue from both species contained higher deposit feeders to changes of environmental contami- metal concentrations than skeletal material. For most nation. The transfer of metals from prey to predator will elements, the highest concentrations were measured in also depend on the type of metal and its bioavailability individuals from the site adjacent to the source of con- in the prey ,Viarengo and Nott 1993; Nott and Nico- tamination. For several elements, signi®cant di€erences laidou 1994). In N. armata, both tissues analysed are were only apparent in the digestive tract, suggesting that part of the digestive system and showed a strong signal this tissue may be the most sensitive indicator of envi- of elevated Pb at the sites adjacent to the pollutant ronmental contamination. However, signi®cant di€er- source, indicating that Pb contamination is moving up ences in Pb concentrations were also detected in the food chain. skeleton. Echinoderms are known to accumulate some The gammaridean amphipod, P. walkeri, is amongst elements such as Pb in their skeleton ,Auernheimer and the most abundant Antarctic benthic species in the depth Chinchon 1997; Temara et al. 1997, 1998b). range of 0±15 m, and frequent among red algae in The asteroid, N. armata, is an active predator and shallow water ,Sagar 1980). Because it is highly mobile, scavenger ,McClintock 1994) known to feed on bivalves, it can avoid unfavourable conditions. In this study, Pb including Limutula hodgsoni ,Dearborn 1977) and was the only element that accumulated signi®cantly in Adamussium colbecki ,Berkmann 1990). If the major individuals kept in enclosures close to the source of 213

Table 3 Ratios of mean concentrations of lead, copper and zinc in tissues of various species collected at contaminated ,Brown Bay site 1 BB1) and reference sites ,Shannon Bay SH and O'Brien Bay OB), or transplanted to sites a and b within Brown Bay Species Tissue Sites Lead Copper Zinc

Bivalve Laternula elliptica Kidney BB1/SH 7.0 2.1 1.1 Gills 19.8 1.3 1.1 Muscle 8.2 2.3 1.4 Heart urchin Abatus sp. Skeleton BB1/OB A. nimrodi 8.3 1 1.1 A. ingens 7.41.5 0.8 Digestive tract A. nimrodi BB1/SH 10.0 2.1 1.2 BB1/OB 47.0 1.7 2.6 A. ingens BB1/SH 14.8 2.5 1.5 BB1/OB 43.8 3.0 2.4 Star®sh Notasterias armata Pyloric caeca BB1/SH 4.8 1.0 1.0 BB1/OB 3.5 1.1 0.9 Stomach BB1/SH 3.7 1.41 BB1/OB 6.1 1.6 1.3 Gammarid Whole body BBa/BBb 3.8 1 0.9 pollution, most likely because it is the predominant Comparison of various species and identi®cation contaminant. of sources of contamination Although size and sex of individuals could partly account for some of the variations of metal concentra- Among the species studied, the ®lter-feeding bivalve tions observed in all species, statistical tests ,ANOVA) Laternula elliptica was the most sensitive in re¯ecting the showed that these parameters have no in¯uence. The contamination since the bioaccumulation of Pb and Cu di€erences in metal concentrations in biota were thus in its tissues was generally the highest ,Table 3). mainly due to changes of concentrations in the envi- The deposit feeder Abatus sp. also re¯ects very well ronment and were consistent with a gradient of con- the contamination, especially in soft tissue such as the tamination from Brown Bay through Shannon and digestive tract. Newcombe Bays to O'Brien Bay. With the predatory species of star®sh, the results The di€erent species tested in this investigation indicated that the contamination is moving up the present various advantages linked to their characteris- food chain but the bioaccumulation was not occur- tics. ring to the same extent as in bivalves and heart ur- The bivalve Laternula elliptica has many biological chins. and ecological features that are similar to those of In the bivalve, the highest increase of Pb in individ- mussels and oysters used in biomonitoring programmes uals collected adjacent to the pollutant source compared such as the Mussel Watch programme ,Goldberg et al. with reference locations, occurred in the gills while the 1978; Lauenstein et al. 1990). Results from this study increases of Cu were highest in kidney and muscle demonstrated that Laternula elliptica might also be ,Table 3). Increases of copper were also signi®cant in the suitable for detecting environmental changes of metal digestive gland of heart urchins. Gammarids kept in concentrations. the water column also accumulated signi®cant amounts The heart urchin Abatus sp. can be useful for moni- of the predominant contaminant, Pb, over a short period toring e€ects of remediation over di€erent time scales, as of time. proposed for the star®sh, Asterias rubens ,Temara et al. These results lead to the hypothesis that Pb may be 1998a). Indeed, both tissues tested responded to Pb mainly present in the water column ,dissolved phase contamination and could be considered as complemen- or suspended particulates) while Cu could be mainly tary temporal biological indicators. associated with sediments. The identi®cation of a The asteroid N. armata should be e€ective for mon- particular source of contaminant is a major advantage itoring the changes and recovery of an ecosystem after of the comparative use of di€erent biological indica- remediation because metal concentrations should re¯ect tors ,Phillips and Rainbow 1988, 1993). But our hy- those in species in its foraging range. pothesis should be con®rmed ,1) by more detailed The amphipod P. walkeri was deployed successfully information on the relative ability of each species to in enclosures and this opens the possibility of targeted accumulate metals from various sources in their dif- monitoring, designed to de®ne ®ne-scale spatial and ferent tissues, and ,2) by information on additional temporal patterns of environmental contaminants. Bio- elements, as it is dicult to draw conclusions related monitoring that can provide such rapid feedback is a to the copper contamination considering its biological very useful tool. essentiality. 214

Comparison with species from lower latitudes species, structure and survival. Cambridge University Press, Cambridge, pp 142±151 Ahn IY, Lee S, Kim K, Shim J, Kim D ,1996) Baseline heavy metal The concentrations of Cd, Cu and Zn in gills of concentrations in the Antarctic clam, Laternula elliptica in Laternula elliptica from reference sites were compared Maxwell Bay, King George Island, Antarctica. Mar Pollut Bull with those of other Antarctic and temperate bivalves 32:592±598 reported in Viarengo et al. ,1993). Concentrations in Auernheimer C, Chinchon S ,1997) Calcareous skeletons of sea Antarctic species ,Laternula elliptica and Adamussium urchins as indicators of heavy metals pollution, Portman Bay, Spain. Environ Geol 29:78±83 colbecki) were higher for Cd ,about tenfold) and lower Bargagli R, Nelli L, Ancora S, Focardi S ,1996) Elevated cadmium for Cu ,about twofold) than in temperate species ,Pecten accumulation in marine organisms form Terra Nova Bay jacobeus and Mytilus galloprovincialis). ,Antarctica). Polar Biol 16:513±520 Results obtained with the gammarid Paramoera Berkmann PA ,1990) The population biology of the Antarctic scallop, Adamussium colbecki ,Smith 1902) at New Harbor, walkeri showed a similar trend. Most concentrations Ross Sea.In: Kerry KR, Hempel G ,eds) Antarctic ecosystems: reported for amphipods from di€erent marine regions of ecological change and conservation. Springer, Berlin Heidelberg the world were within a range of 0.1±3.0 ppm for Cd and New York, pp 281±288 30±90 ppm for Cu ,Petri and Zauke 1993). Compared to Berkmann PA ,1992) The Antarctic marine ecosystem and humankind. Rev Aquat Sci 6:295±333 these species, Paramoera walkeri had high levels of Cd Bryan GW, Langston WJ, Hummerstone LG, Burt GR ,1985) A ,5.2 ppm in this study and 7.2 ppm in Bargagli et al. guide to the assessment of heavy metal contamination in estu- 1996) and low levels of Cu ,26.3 ppm). aries using biological indicators. Occas Publ Mar Biol Assoc The di€erences observed between similar species from UK 4:1±92 di€erent latitudes may re¯ect variations of environ- Dearborn JH ,1977) Foods and feeding characteristics of antarctic asteroids and ophiuroids. In: Liano GA ,ed) Proceedings of the mental enrichment in polar and temperate ecosystems or third SCAR symposium on Antarctic biology, Houston, Texas. be indicative of di€erences in metal requirements for Gulf, Houston, pp 293±326 certain species ,Petri and Zauke 1993; Viarengo et al. Depledge MH, Rainbow PS ,1990) Models of regulation and ac- 1993; Bargagli et al. 1996). cumulation of trace metals in marine invertebrates. Comp Biochem Physiol 97C:1±7 Deprez PP, Arens M, Locher H ,1999) Identi®cation and assess- Conclusions ment of contaminated sites at Casey Station, Wilkes Land, Antarctica. Polar Rec 35:299±316 Dubois P, Jangoux M ,1985) The microstructure of the asteroid This study showed that the environmental contamina- skeleton ,Asterias rubens). In: Keegan BF, O'Connor BDS ,eds) tion of nearshore waters induced accumulation of heavy Echinodermata. Balkema, Rotterdam, pp 507±512 metals in biota and that concentrations in the di€erent Duquesne S, Riddle M, Schulz R, Liess M ,2000) E€ects of con- taminants in the Antarctic environment ± potential of the species of invertebrates were consistent with a gradient gammarid amphipod crustacean Paramoera walkeri as a bio- of contamination. logical indicator for Antarctic ecosystems based on toxicity and The species selected were shown to be good candi- bioacccumulation of copper and cadmium. Aquat Toxicol dates for biological indicators for future monitoring of 49:131±143 heavy-metal contamination. However, our ®ndings re- Farrington JW, Goldberg ED, Risebrough ,1983) United States mussel watch 1976±1978 ± an overview of the trace metal, enforce the need for controlled toxicokinetic studies for DDE, PCB, hydrocarbon and arti®cial radionuclide data. a better understanding of the characteristics of bioac- Environ Sci Technol 17:490±496 cumulation in species proposed for ®eld biomonitoring, Goldberg ED, Bowen VT, Farrington JW, Harvey G, Martin JH, as they have various functional roles in the marine Parker PL, Risebrough RW, Robertson W, Schneider E, Gamble E ,1978) The Mussel Watch. Environ Conserv 5: ecosystem. Such studies were performed for the crusta- 101±125 cean Paramoera walkeri ,Duquesne et al. 2000) and are Langston WJ, Zhou MJ ,1987) Cadmium accumulation, distribu- currently underway for the bivalve, Laternula elliptica. tion and metabolism in the gastropod Littorina littorea ± the role of metal-binding proteins. J Mar Biol Assoc UK 67: Acknowledgements This study was supported by a grant from the 585±601 Australian Antarctic Science Advisory Committee. Drs. Matthias Lauenstein GG, Robertson A, O'Connor TP ,1990) Comparison of Liess and Ian Snape are thanked for discussions leading to im- trace metal data in mussels and oysters from a mussel watch provements in this study and related to this manuscript. We would program of the 1970s with those from a 1980s program. Mar also like to thank the Australian Antarctic Division, as well as Pollut Bull 21:440±447 S. Kratzman, M. Greve, C. King and the sta€ of Casey station. Liess M, Duquesne S ,1997) Fate and ecotoxicological e€ect of A. Tabor, P. Goldsworthy and J. Stark are thanked for their as- the contaminants of Thala Valley tip site, Casey, Antarctica. sistance with diving and specimen collection. We are also grateful ANTDIV 2228. Australian Antarctic Division, Hobart, to Drs A. Greig ,University of Queensland) and M. Wu ,AGAL, Tasmania Sydney) for the ICP-MS analyses. Long ER, McDonald DD, Smith SL, Calder FD ,1995) Incidence of adverse biological e€ects within ranges of chemical concen- trations in marine and estuarine sediments. Environ Manage 19:81±97 References McClintock JB ,1994) Trophic biology of Antarctic shallow-water echinoderms. Mar Ecol Prog Ser 111:191±202 Ahn IY ,1997) Feeding ecology of the Antarctic lamellibranch Moore PG, Rainbow PS, Hayes E ,1991) The beach-hopper Laternula elliptica ,Laternulidae) in Marian Cove and vicinity, Orchestia gammarellus ,Crustacea: Amphipoda) as a biomoni- King George Island, during one austral summer. In: Battaglia tor for copper and zinc: North Sea trials. Sci Total Environ B, Valencia J, Walton DWH ,eds) Antarctic communities: 106:221±238 215

Nott JA, Nicolaidou A ,1994) Variable transfer of detoxi®ed Temara A, Nguyen QA, Hogarth AN, Warnau M, Jangoux M, metals from snails to hermit crabs in marine food chains. Mar Dubois P ,1997) High sensitivity of skeletogenesis to Pb in the Biol 120:369±377 asteroid Asterias rubens ,Echinodermata). Aquat Toxicol Petri P, Zauke GP ,1993) Trace metals in crustaceans in the 40:1±10 Antarctic Ocean. Ambio 22:529±536 Temara A, Aboutboul P, Warnau M, Jangoux M, Dubois P Phillips DJF ,1980) Quantitative aquatic biological indicators. ,1998a) Uptake and fate of lead in the common asteroid As- Applied Sciences, London terias rubens ,Echinodermata). Water Air Soil Pollut 102:201± Phillips DJH, Rainbow PS ,1988) Barnacles and mussels as bio- 208 monitors of trace elements ± a comparative study. Mar Ecol Temara A, Skei JM, Gillan D, Warnau M, Jangoux M, Dubois P Prog Ser 49:83±93 ,1998b) Validation of the asteroid Asterias rubens ,Echinoder- Phillips DJH, Rainbow PS ,1993) Cosmopolitan biomonitors of mata) as a bioindicator of spatial and temporal trends of Pb, trace-metals. Mar Pollut Bull 26:593±601 Cd and Zn contamination in the ®eld. Mar Environ Res Rainbow PS ,1985) The biology of heavy metals in the sea. Int 45:341±345 J Environ Stud 25:195±211 Van den Brink NW ,1997) Directed transport of volatile orga- Rainbow PS, White SL ,1989) Comparative strategies of heavy nochlorine pollutants to polar regions: the e€ect on the con- metal accumulation by crustaceans: zinc, copper and cadmium tamination pattern of Antarctic seabirds. Sci Total Environ in a decapod, an amphipod and a barnacle. Hydrobiologia 198:43±50 174:245±262 Viarengo A ,1985) Biochemical e€ects of trace metals. Mar Pollut Risebrough RW, Walker W, Schmidt TT, De Lappe DW, Connors Bull 16:153±158 CW ,1976) Transfer of chlorinated biphenyls to Antarctica. Viarengo A, Nott JA ,1993) Mechanisms of heavy metal cation Nature 264:738 homeostasis in marine invertebrates. Comp Biochem Physiol Sagar PM ,1980) Life cycle and growth of the Antarctic gammarid 104C:355±372 amphipod Paramoera walkeri ,Stebbing, 1906). J R Soc N Z Viarengo A, Canesi L, Mazzucotelli A, Ponzano E ,1993) Cu, 10:259±270 Zn and Cd content in di€erent tissues of the Antarctic scal- Serra R, CarpeneÁE, Marcantonio AC, Isani G ,1995) Cadmium lop Adamussium colbecki: role of metallothionein in heavy accumulation and Cd-binding proteins in the bivalve Scapharca metal homeostasis and detoxication. Mar Ecol Prog Ser 95: inaequivalvis. Comp Biochem Physiol 111C:165±174 163±168 Snape I, Riddle MJ, Stark JS, Cole CM, King CK, Duquesne S, Warnau M, Ledent G, Temara A, Jangoux M, Dubois P ,1995) Gore DB ,in press) Management and remediation of contami- Experimental cadmium contamination of the echinoid Para- nated sites at Casey Station, Antarctica. Polar Rec centrotus lividus: in¯uence of exposure mode and distribution of Stebbing ARD, Dethlefsen V, Thurberg F ,1992) Report on the ICES the metal in the organism. Mar Ecol Prog Ser 116:117±124 North Sea seagoing workshop. Water Sci Technol 24:21±23 Zauke GP, Von Lemm R, Meurs HG, Butte W ,1995) Validation Sullivan PA, Robinson WE, Morse MP ,1988) Subcellular distri- of estuarine gammarid collectives ,Amphipoda: Crustacea) as bution of metals within the kidney of the bivalve Mercenaria biomonitors for cadmium in semi-controlled toxicokinetic mercenaria ,L.). Comp Biochem Physiol 91C:589±596 ¯ow-through experiments. Environ Pollut 90:209±219