Cadmium-109 Internal Kinetics in Diamond Sturgeon Acipenser gueldenstaedtii are Strongly Influenced by Salinity, Exposure Pathway and History.

Cadmium-109 whole-body and internal biokinetics were experimentally investigated in critically endangered diamond sturgeon Acipenser gueldenstaedtii after uptake from water or food, in fresh (FW) and brackish (BW; 9‰) salinities typical of the Caspian Sea. Whole-body rates of uptake of 109Cd from water and subsequent depuration were quantified over 14 and 28 days, respectively. Uptake was greater in FW than BW by a factor of 2.4, but depuration rates were similar in both salinities. In contrast, for the dietary (chironomid) exposure pathway 109Cd assimilation efficiencies (AEs) were higher in BW (13%) compared to FW (9.5%). Head (including gills) or digestive tract were major repositories of 109Cd following aqueous and dietary exposures, respectively, including both uptake and depuration phases. The point-of-entry of 109Cd into the body was also a major and persistent determiner of its subsequent internal distribution. For aqueous exposures, the internal distributions of 109Cd changed appreciably during depuration with increased activity concentrations in some body components, which again varied with salinity. Increased salinity appreciably enhanced the percentage distributions and activity concentrations of 109Cd in the liver, kidney and digestive tract, which are typically most pathologically altered by elevated Cd exposure. For dietary exposure, increased salinity also enhanced 109Cd activity concentrations in most body components. The results repeatedly indicate the important role of salinity in the whole-body and internal biokinetics of 109Cd in A. gueldenstaedtii, a representative of both a phylogenetically distinct and most endangered family of fishes.

Body temperature and time of day both affect nocturnal lizard performance: An experimental investigation.

The locomotor performance of reptiles is profoundly influenced by temperature, but little is known about how the time of day when the animal is usually active may influence performance. Time of day may be particularly relevant for studies on nocturnal reptiles that thermoregulate by day, but are active at night when ambient temperatures are cooler. If selection favours individuals that match their performance to activity times, then nocturnal species should perform better during the night, when they are normally active, than during the day. To test this hypothesis, we investigated how the time of day and body temperature affected the locomotor performance of adult females of the velvet gecko (Amalosia lesueurii). We measured the sprint speeds, running speeds and number of stops of 43 adult females at four different body temperatures (20, 25, 30 and 35 °C) during the day and at night. At night, sprint speeds were higher at 20 and 35 °C but sprint speeds were similar at 25 and 30 °C. By day, sprint speed increased with body temperature, peaking at 30 °C, before declining at 35 °C. However, gecko speeds over 1 m was higher at night at all four test temperatures than by day. Number of stops showed broadly similar patterns and females stopped almost twice as often on the racetrack during the day than they did at night. Furthermore, the thermal breadth of performance differed depending on when geckos were tested. Our results demonstrate that both body temperature and the time of day affects the behaviour and locomotor performance of female velvet geckos, with geckos running faster at night, the time of day when they are usually active. This study adds to evidence that both body temperature and the time of day are crucial for estimating the performance of ectotherms and evaluations and predictions of their vulnerability to climate warming should consider the context of laboratory experimental design.

Maternal transfer of trace elements to shark eggs and their dietary assimilation efficiencies.

Dogfish (Scyliorhinus canicula) transferred trace elements (110Ag, 109Cd, 54Mn and 75Se) from their diet to eggs, and their components (yolk and embryo, case and jelly) at greatly varying rates. Trace element levels in eggs showed positive linear relationships (p < 0.001; r2-0.83-0.91) with their cumulative rates of maternal ingestion over 61 days (maternal-to-egg transfer rates: mTFs). These mTFs varied by 2-3 orders of magnitude, with 54Mn > 110Ag > 75Se > 109Cd, and their range encompassed those previously measured for 60Co, 65Zn, 241Am and 134Cs. For six of the eight trace elements, their mTFs were significantly influenced (p < 0.05; r2 = 0.72) by both their dietary assimilation efficiency and their location within the egg (case). In contrast, both 110Ag and 54Mn greatly exceeded the mTFs predicted by this multiple regression model by one and 2-3 orders of magnitude, respectively, and were predominantly transferred to the egg case. Among elements, contrasting rates of transfer and percentage distributions in egg components imply differing ecotoxicological and radiological detriments to the developing embryo.

Internal distributions of a radio-element array in cartilaginous and bony marine fishes: Different and heterogeneous.

This experimental study determined internal distributions of an array of radio-elements (54Mn, 60Co, 65Zn, 134Cs, 241Am, 109Cd, 110mAg, 75Se and 51Cr) accumulated from seawater by three chondrichthyan fish species (Scyliorhinus canicula (dogfish), Raja undulata (undulate ray) and Torpedo marmorata (spotted torpedo)) and three teleost species (Scophthalmus maximus (turbot), Sparus aurata (seabream) and Dicentrarchus labrax (seabass)). The study tested the hypothesis that the chondrichthyan (cartilaginous) fish taxon and teleost (bony) fish taxon have different patterns of bioaccumulation of these radio-elements in six body components (head, digestive tract, liver, kidneys, skin and muscle), consistent with their long period of evolutionary divergence. Comparisons of body component CFs between the two taxa for each radio-element and the full array of radio-elements showed highly significant differences (p ≤ 0.001) between cartilaginous and bony fishes in each body component, confirming the existence of a strong and pervasive phylogenetic signal; however, the subset of radio-elements most determinant of these differences were unique for each body component. Partitioning between the three individual bony species and the three individual cartilaginous species also occurred repeatedly among their body components, particularly for bony fishes. Distributions of these radio-elements among body components were typically highly heterogeneous for both fish taxa.

Physico-chemical and key metal data for surface waters and sediments of the Sydney and Hawkesbury estuaries, Australia.

This article contains general physico-chemical data (salinity, pH, redox potential, temperature, dissolved oxygen, suspended particulate matter (SPM), dissolved organic carbon and chlorophyll a concentrations) for surface waters at 15 near-pristine sites in the Hawkesbury Estuary and 24 sites (encompassing a wide range of metal contamination) in the highly urbanized Sydney Estuary, south-eastern Australia. Data on concentrations of five key metals (cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb) and zinc (Zn)) in filtered (<0.2 µm) surface water, suspended particulate matter (>0.2 µm) and surface sediments (<2 mm) at each study site are also provided. The concentrations of Cd, Cr, Cu, Pb and Zn in SPM and sediment at each site were normalised for aluminium (Al) concentration (e.g. Cd/Al), to account for natural variation in particle size and mineralogy. Enrichment factors (EFs) were calculated from these data by dividing the mean metal concentration at each site in the Sydney Estuary, for each environmental matrix (i.e., filtered water, SPM and sediment), by its mean baseline metal concentration from near-pristine reference sites in the adjacent Hawkesbury Estuary. A thorough knowledge of the general physico-chemistry and key metal concentrations in surface waters and sediments in the Sydney Estuary provide a baseline to assess anthropogenic change and better manage estuarine/marine ecosystems.

The euryhaline pygmy mussel, Xenostrobus securis, is a useful biomonitor of key metal contamination in the highly urbanised Sydney Estuary, Australia.

This study critically evaluated the native pygmy mussel (Xenostrobus securis) as a biomonitor of the key metal contaminants in the highly urbanised Sydney Estuary, south-eastern Australia. Five metals (Cd, Cr, Cu, Pb and Zn) were identified as key contaminants, based on their enrichment factors (EFs) in the whole soft tissue of X. securis at 24 sampling sites, relative to baseline values from near-pristine reference sites in the adjacent Hawkesbury Estuary. Inverse relationships established between mussel size (dry tissue weight) and tissue concentrations of each metal were used to reduce variance (by 4-fold) among individuals; gender and reproductive status had no significant (p > 0.05) effect on tissue metal concentrations in X. securis. Metal concentrations in three environmental matrices - filtered (<0.2 µm) surface water (operationally defined as the dissolved/colloidal phase), suspended particulate matter (SPM; >0.2 µm) and surface sediment (<2 mm particle size), which are most relevant to a suspension-feeding estuarine bivalve, were also determined at each sampling site. For each of the five metals, highly significant (p < 0.01) positive linear regressions were established between metal EFs for mussel tissue and each environmental matrix. Metals in surface sediment and SPM explained 80-91% and 81-90%, respectively, of the variability in metal concentrations in mussel tissue, with filtered surface water explaining 74-86%. Cumulative mussel tissue EFs of all five metals, when regressed against each environmental matrix, showed that surface sediment concentrations explained 93% of their variability between sites, SPM 94% and filtered surface water 87-90%. Hence, X. securis very closely reflects the metal concentrations in its aquatic environment. The study provides a quality-assured benchmark of key metal contamination in the Sydney Estuary, and an appropriate methodology that may be used to discern any changes in metal contaminant status using X. securis.

Radioecological aftermath: Maternal transfer of anthropogenic radionuclides to shark progeny is sustained and enhanced well beyond maternal exposure.

Cartilaginous dogfish Scyliorhinus canicula continued to transfer four anthropogenic radionuclides (65Zn, 60Co, 134Cs and 241Am) to their eggs for over six months, after two months of continued maternal exposure to radio-labelled food. Unexpectedly, rates of radionuclide transfers to eggs and their yolk & embryo during maternal depuration were equivalent for 60Co and 241Am, or even enhanced for 65Zn and 134Cs by factors of c.200-350%, over two-three months, compared to their maximal transfer rates at the end of the maternal uptake phase. These rates of maternal transfer of radionuclides to yolk & embryo were positively associated with their previously determined efficiencies of assimilation (AE) from ingested radio-labelled food. Thus progeny may be more exposed via maternal transfer to those radionuclides which have greater rates of maternal assimilation from food. As maternal depuration continued beyond 60-80 up to 180-200 days the transfers of all four radionuclides to eggs did diminish but were still substantial at mean values of 18% for 241Am, 17% for 134Cs and 9 and 8% for 60Co and 65Zn, respectively. In the yolk & embryo the mean rates of transfer over this period were further reduced for 241Am (13.5%), 60Co (2.5%) and 65Zn (5.8%), but were still appreciable for 134Cs at 56%. These results for S. canicula have demonstrated a potential enhanced radiological risk of extended duration due to the particular biokinetics of maternal transfer in this species. This study draws further attention to the current paucity of knowledge about the maternal: progeny transfer pathway, particularly in the context of the known heightened radio-sensitivity of early life stages in fish and other vertebrates, compared to later life stages.

Accumulation and transport behaviour of 241americium, 60cobalt and 134cesium by eggs of the spotted dogfish Scyliorhinus canicula.

An experimental study examined the 96-h net influx from seawater of the anthropogenic radionuclides (241)Am, (60)Co and (134)Cs through the egg-case of the spotted dogfish Scyliorhinus canicula. Net influx directly through the wall of the egg-case was greatest for (134)Cs, then (241)Am and lastly (60)Co. Within the egg-case wall itself the measured concentration factors (CFs) and their gradients in the external, median and internal layers showed that for both (241)Am and (60)Co they were >10(3) in the external layer and declined by an order of magnitude in the interior layer. In contrast (134)Cs had a CF of only about three in the external layer which declined by a factor of 2 towards the two more internal layers of the egg-case. The egg-case apertures, that open within the prehatching stage of embryological development, significantly (P<0.05) increased the net influx of (241)Am and (60)Co to the interior of the egg-case, although their water concentrations were still lower than those in the labelled seawater bath. In contrast, the aperture did not increase the net influx of (134)Cs whose water concentrations equilibrated with those in seawater. Together these results indicate that the egg-case wall is very permeable to (134)Cs, representing little barrier to its movement, and hence consistent with the lack of importance of the aperture in determining its internal water concentrations in the egg-case. In contrast, (241)Am and (60)Co show much higher rates of accumulation by the egg-case, consistent with the measured reduced permeability of its wall, and therefore giving greater prominence to its aperture in the net transfer of these two radionuclides to the egg case's interior. The presence of the embryo within its egg-case did not significantly (P>0.05) affect the rates of influx of radioisotopes, with the exception of an interactive effect for (60)Co with the egg-case aperture (P<0.05). The CF of only (241)Am in the embryo itself relative to the external seawater concentration was significantly (P<0.05) enhanced by the presence of the aperture.

Experimental comparison of the bioaccumulation of anthropogenic radionuclides by egg and juvenile life stages of a small shark.

This study compared the bioaccumulation of anthropogenic nuclides (65Zn, 134Cs, 60Co and 241Am) between the egg and juvenile life stages of a small shark (Scyliorhinus canicula), based on previously published experimental data. Rates of accumulation over 15 days were derived and summed for the transfer pathways which were specific to these two life stages. Radionuclide transfers to the egg and its embryo & yolk were quantified for i) the maternal pathway following her uptake of radionuclides via food and seawater and ii) from seawater following its oviposition. For the juvenile, the transfer of radionuclides were measured for aqueous & dietary pathways. The results show that, compared to juveniles, eggs have equivalent rates of accumulation of 65Zn and 134Cs but enhanced accumulation of 241Am by a factor of five and of 60Co by two orders of magnitude. The radiological exposure of the embryo due to radionuclides maternally transferred to the embryo & yolk is also enhanced for the alpha-emitting 241Am. This enhanced accumulation of 241Am and 60Co, as well as the equivalent accumulation of 65Zn and 134Cs, suggest greater likely vulnerability to radiation damage in eggs as compared to juveniles. Radiological dose assessment confirmed highest doses to the egg which is predominantly due to accumulated 241Am.

Radionuclide biokinetics in the Russian sturgeon and phylogenetic consistencies with cartilaginous and bony marine fishes.

The biokinetics of eight radionuclides (241Am, 109Cd, 134Cs, 75Se, 54Mn, 110mAg, 65Zn, 60Co) absorbed from the aquatic medium by juvenile Russian sturgeon (Acipenser gueldenstaedtii) were experimentally determined in fresh (0.42‰) and brackish (9.0‰) waters, of a similar salinity range to the Caspian Sea, and in conjunction with chemical speciation modelling. Uptake and loss rate constants were determined for each radionuclide for a 14 day exposure at each salinity and during 28 days of exposure to radionuclide-free conditions. Whole body (wet): water concentration factors (CF) achieved over 14 days for these eight radionuclides were used in a comparison with the same radionuclide CFs previously determined experimentally for six species of marine teleosts and chondrichthyans, to further test a phylogeny-based model of multi-nuclide bioaccumulation based on marine chordates. Multivariate analyses (multidimensional scaling and hierarchical clustering) identified the relative affinities among these taxa and also those radionuclides which distinguished most between them, in their differing CFs. They consistently showed that sturgeon aggregated as a group, which was also slightly differentiated with salinity. Sturgeon were distinguished from all teleosts and chondrichthyans but were more dissimilar from chondrichthyans than teleosts, in accordance with sturgeon's different periods of divergence from them in evolutionary time. Variable salinity among experiments may also cause changes in radionuclide bioaccumulation due to variations in (i) bioavailability (ii) osmolarity, and (iii) competitive inhibition of a radionuclide's bioaccumulation by its stable analogue or metabolic model. Their potentially confounding effects on these patterns of radionuclide CFs among taxa were critically evaluated for those radionuclides which discriminated most between sturgeon and teleosts or chondrichthyans. Bioavailability, osmolarity and competitive inhibition effects were identified among salinity treatments, however they were not appreciable enough to override the phylogeny-based signal. The results of this study are thus consistent with a phylogeny-based model of radionuclide bioaccumulation by marine chordates being valid for a fish species living in lower salinity regimes.

Comparison of the bioaccumulation from seawater and depuration of heavy metals and radionuclides in the spotted dogfish Scyliorhinus canicula (Chondrichthys) and the turbot Psetta maxima (Actinopterygii: Teleostei).

The bioaccumulation of selected heavy metals and radionuclides ((241)Am, (109)Cd, (57)Co, (51)Cr, (134)Cs, (54)Mn and (65)Zn) from seawater was experimentally compared in the Chondrichthyan Scyliorhinus canicula (spotted dogfish) and the Actinopterygian Teleost Psetta maxima (turbot), of comparable size, age and benthic feeding habits. The speciation of these elements in seawater (salinity 38 per thousand, pH 8.1, temperature 16.5 degrees C) was also calculated to determine their potential bioavailability. The uptake rates, measured over 14 days, varied greatly among isotopes and between species. Concentration factors (CFs) in P. maxima varied 5-fold between ca. 0.2 for (51)Cr and 2.5 for (65)Zn and (134)Cs, whereas in S. canicula they varied by a much greater factor of 350, with CFs for (51)Cr and (241)Am ranging from ca. 0.4 to 140, respectively. With the exception of (134)Cs, all radiotracers were accumulated at a faster rate in S. canicula than in P. maxima, particularly for (241)Am and (65)Zn where the CFs attained during the uptake phase were, two and one order of magnitude greater in S. canicula, respectively. In contrast, (134)Cs reached a CF of about 2.5 in P. maxima, which was 5-fold greater than in S. canicula. Patterns of loss from the experimental depuration phase over 29 days showed greater similarities between species, compared to the uptake phase that highlighted the greater differences between elements. The distributions of these seven radioisotopes among six body components indicated that between the two species the skin of the dogfish displayed a greater bioaccumulation potential, particularly for (241)Am, (57)Co and (65)Zn. However (65)Zn was also distinctive from (241)Am and (57)Co in its pattern of bioaccumulation in dogfish, with its other body components attaining concentrations of (65)Zn that were comparable to the levels found in its skin. The heightened uptake of (134)Cs in turbot was characterised by a more even percentage distribution among its tissues compared to (241)Am and (57)Co, but every tissue of turbot had a higher concentration of (134)Cs, compared to dogfish, particularly the muscle and liver. The elevated uptake rates and higher CFs for most radioisotopes indicate that S. canicula is more susceptible than P. maxima to exposure and contamination by these metals and radionuclides in seawater. These experimentally-determined differences between dogfish and turbot in their bioaccumulation characteristics were assessed against a set of criteria erected to evaluate the working hypothesis that they were taxonomically based. The outcomes of this initial assessment were supportive of this hypothesis that warrants further investigation.

Bioaccumulation of heavy metals and radionuclides from seawater by encased embryos of the spotted dogfish Scyliorhinus canicula.

Encased embryos of spotted dogfish Scyliorhinus canicula absorbed six radio-isotopes (241Am, 109Cd, 57Co, 134Cs, 54Mn and 65Zn) directly from seawater during short-term experimental exposure, demonstrating the permeability of the egg-case to these contaminants. Embryo to water concentration factors (CFs) ranged from 0.14 for 134Cs to 7.4 for 65Zn. The 65Zn and 57Co CFs increased exponentially with embryo length, whereas the CF for 109Cd declined with length. Among different components of the encased embryo the egg case was the major repository (69-99%) of all six radio-isotopes that were distributed throughout its wall. Egg-case CFs were as high as 10(3) for 57Co and 65Zn, making it the major source of gamma radiation exposure to the embryo and potentially of radio-isotopes for continued absorption by the embryo, following the uptake phase of the experiment. The patterns of uptake by the egg-case approximated linearity for most isotopes and loss rates were isotope-specific; egg-case biokinetics were not greatly affected by the viability of the contained embryo. Within the embryo initial data on radio isotopic distribution show that the skin is their major site of uptake, as previously demonstrated for juveniles.

Patterns of metal accumulation in osteoderms of the Australian freshwater crocodile, Crocodylus johnstoni.

The concentrations of 15 metals were measured in the osteoderms (dermal bones) of 30 freshwater crocodiles (Crocodylus johnstoni) from a single population in the Lynd River, northeastern Australia (17 degrees 50'S, 144 degrees 20'E), that were well characterised with respect to site fidelity, reproductive status and age. Multiple linear regression analyses were used to determine the effects of crocodile size (snout-vent length, 25-128 cm), age (0.7-62.7 years), gender (male or female) and reproductive status (sexually mature or immature) on osteoderm metal concentrations. Gender and reproductive status were not significant (P > 0.05) co-predictors of the osteoderm concentration of any metal. In contrast, size, age and osteoderm calcium concentration were highly significant (P < 0.001) systematic predictors of the osteoderm concentrations of all metals, except Na and K. Osteoderm metal concentrations were inversely related (P < 0.001) to both size (r(2) = 0.52-0.92) and age (r(2) = 0.52-0.84), but positively related (P < 0.001) to osteoderm calcium concentration (r(2) = 0.67-0.92). Relative to calcium concentration, the rates of metal accumulation in the osteoderms of C. johnstoni were inversely related to the solubility constant (log K(sp)) of the metal as a phosphate; however this relationship was not linear. This finding is consistent with that previously established for the flesh of freshwater bivalves, which like the crocodilian osteoderm, have a calcium phosphate repository in the form of extracellular granules. The constancy of this relationship between rate of metal accumulation and relative solubility for calcium phosphate deposits, despite contrasting Ca accumulation regimes and taxonomic dissimilarity, points to a potential underlying principle that warrants investigation in a greater range of biota. The implications for using the osteoderms of C. johnstoni as an indicator of metal levels in freshwater ecosystems are also discussed.

Maternal transfer of anthropogenic radionuclides to eggs in a small shark.

Maternal transfer of radionuclides to progeny is one of the least known sources of contamination in marine biota and more information is needed to assess its radiological significance. A radiotracer study on spotted dogfish, Scyliorhinus canicula, evaluated the hypothesis that four anthropogenic radionuclides (Cobalt-60, Zinc-65, Americium-241 and Cesium-134) could be maternally transferred to eggs and each of their major components during maternal ingestion of radiolabelled food. The linear regressions between cumulative radioactivity that had been maternally ingested and the level in subsequently laid eggs were used to derive maternal-to-egg transfer factors (mTFs). These maternal transfers varied over an order of magnitude and were ranked (134)Cs > (65)Zn > (60)Co > (241)Am. This ranking was the same as their relative assimilation efficiencies in radiolabelled food consumed by adults. Among these four radionuclides the potential radiological exposure of embryos is accentuated for (65)Zn and (134)Cs due to their predominant transfer to egg yolk where they are available for subsequent absorption by the embryo as it develops prior to hatching from the egg capsule. Thus, for cartilaginous fish like shark, the potential radioecological consequences of a pulsed release of these radionuclides into the marine environment may extend beyond the temporal duration of the release.

Insights into the mechanisms of copper tolerance of a population of black-banded rainbowfish (Melanotaenia nigrans) (Richardson) exposed to mine leachate, using 64/67Cu.

This study investigates the mechanisms of copper tolerance of a population of black-banded rainbowfish (Melanotaenia nigrans) (Richardson). The population has been exposed to elevated copper concentrations for over 40 years, due to leachate from the Rum Jungle uranium/copper mine. At the time of collection the 96 h EC(50) of exposed [E] fish was 8.3 times higher than that of reference [R] fish. The bioconcentration of 64/67Cu in fish was used to investigate the mechanism of copper tolerance in E fish. Both E and R fish were exposed to low (L(Cu), 30 microg Cu l(-1)) and elevated (E(Cu), 300 microg Cu l(-1)) copper concentrations for 24 and 48 h, respectively. Radioactivity was measured at seven or eight time points in four tissue sections: head (including gills, heart and brain), internal organs (including gastrointestinal tract, liver, kidneys and gonads), muscle and whole body. One-compartment bioconcentration models were fit to data and compared using an F-test. Copper concentrations in all tissue sections were significantly (P<0.05) less (up to 50%) in E fish compared with the respective tissue sections of R fish when exposed to both L(Cu) and E(Cu). The exception was copper accumulation in the internal organs, which was not significantly different between E and R fish exposed to E(Cu). The mechanism of copper tolerance was concluded to be reduced copper uptake in the gills, rather than increased binding or elimination. Allozyme electrophoresis was performed to determine if genetic selection had occurred in the E fish population. Allozyme frequencies at the AAT-1 and GPI-1 loci were significantly (P<0.05) different between E and R fish. Heterozygosity was reduced in E fish compared with that of R fish. Collectively these results suggest that genetic selection may have occurred in the E fish population. Consequently, the selection of allozymes less sensitive to copper may be another mechanism of copper tolerance of E fish. This is the first study on the mechanisms of copper tolerance in a wild fish population that has been exposed to elevated copper concentrations. These findings aid the understanding of metal tolerance in fish and emphasise the importance of sample selection and its implication for toxicity testing.

Catchment-specific element signatures in estuarine crocodiles (Crocodylus porosus) from the Alligator Rivers Region, northern Australia.

The concentrations of Na, K, Ca, Mg, Ba, Sr, Fe, Al, Mn, Zn, Pb, Cu, Ni, Cr, Co, Se, U and Ti were determined in the osteoderms and/or flesh of estuarine crocodiles (Crocodylus porosus) captured in three adjacent catchments within the Alligator Rivers Region (ARR) of northern Australia. Results from multivariate analysis of variance showed that when all metals were considered simultaneously, catchment effects were significant (P < or = 0.05). Despite considerable within-catchment variability, linear discriminant analysis (LDA) showed that differences in elemental signatures in the osteoderms and/or flesh of C. porosus amongst the catchments were sufficient to classify individuals accurately to their catchment of occurrence. Using cross-validation, the accuracy of classifying a crocodile to its catchment of occurrence was 76% for osteoderms and 60% for flesh. These data suggest that osteoderms provide better predictive accuracy than flesh for discriminating crocodiles amongst catchments. There was no advantage in combining the osteoderm and flesh results to increase the accuracy of classification (i.e. 67%). Based on the discriminant function coefficients for the osteoderm data, Ca, Co, Mg and U were the most important elements for discriminating amongst the three catchments. For flesh data, Ca, K, Mg, Na, Ni and Pb were the most important metals for discriminating amongst the catchments. Reasons for differences in the elemental signatures of crocodiles between catchments are generally not interpretable, due to limited data on surface water and sediment chemistry of

Diminished metal accumulation in riverine fishes exposed to acid mine drainage over five decades.

Bony bream (Nematalosa erebi) and black catfish (Neosilurus ater) were sampled from the fresh surface waters of the Finniss River in tropical northern Australia, along a metal pollution gradient draining the Rum Jungle copper/uranium mine, a contaminant source for over five decades. Paradoxically, populations of both fish species exposed to the highest concentrations of mine-related metals (cobalt, copper, lead, manganese, nickel, uranium and zinc) in surface water and sediment had the lowest tissue (bone, liver and muscle) concentrations of these metals. The degree of reduction in tissue concentrations of exposed populations was also specific to each metal and inversely related to its degree of environmental increase above background. Several explanations for diminished metal bioaccumulation in fishes from the contaminated region were evaluated. Geochemical speciation modeling of metal bioavailability in surface water showed no differences between the contaminated region and the control sites. Also, the macro-nutrient (calcium, magnesium and sodium) water concentrations, that may competitively inhibit metal uptake, were not elevated with trace metal contamination. Reduced exposure to contaminants due to avoidance behavior was unlikely due to the absence of refugial water bodies with the requisite metal concentrations lower than the control sites and very reduced connectivity at time of sampling. The most plausible interpretation of these results is that populations of both fish species have modified kinetics within their metal bioaccumulation physiology, via adaptation or tolerance responses, to reduce their body burdens of metals. This hypothesis is consistent with (i) reduced tissue concentrations of calcium, magnesium and sodium (macro-nutrients), in exposed populations of both species, (ii) experimental findings for other fish species from the Finniss River and other contaminated regions, and (iii) the number of generations exposed to likely selection pressure over 50 years.

Marine radionuclide transfer factors in chordates and a phylogenetic hypothesis.

Previous radiotracer experiments that compared multi-elemental whole organism: water transfer factors among chondrichthyan and teleost fishes, including an ICRP reference flatfish Psetta maxima, demonstrated distinctive contrasts in their bioaccumulation characteristics, with generally elevated bioaccumulation in chondrichthyans. These results supported a hypothesis that phylogenetic divergence may influence marine radionuclide transfer factors. This notion has been further evaluated in an amphioxus species Branchiostoma lanceolatum, sub-phylum Cephalochordata. This taxon diverged about 800 MYBP from a common ancestor of the teleosts and the chondrichthyans, which in turn diverged from each other around 500 MYBP. Our experimental results indicate that amphioxus is indeed more divergent in its multi-elemental bioaccumulation patterns from teleosts and chondrichthyans than they are from each other, consistent with our hypothesis. The experimental comparisons with the ICRP reference flatfish P. maxima also revealed an unexpectedly enhanced capacity in amphioxus to accumulate all eight tested trace elements from seawater, and for some by more than two orders of magnitude. These results have practical applications for the strategic selection of marine biota for further radioecological investigations to better guarantee the radiological protection of marine biodiversity. Such seemingly anomalous results for understudied biota like amphioxus and chondrichthyans suggest that more effort in marine radioecology be directed to assessing the bioaccumulatory capacities of other phylogenetic groups that have received less attention so far, particularly those that are phylogenetically more remote from commonly investigated taxa and those nominated as ICRP marine reference organisms.

Cobalt-induced genotoxicity in male zebrafish (Danio rerio), with implications for reproduction and expression of DNA repair genes.

Although cobalt (Co) is an environmental contaminant of surface waters in both radioactive (e.g. (60)Co) and non-radioactive forms, there is relatively little information about Co toxicity in fishes. The objective of this study was to investigate acute and chronic toxicity of Co in zebrafish, with emphasis on male genotoxicity and implications for reproductive success. The lethal concentration for 50% mortality (LC(50)) in larval zebrafish exposed (96 h) to 0-50 mg l(-1) Co was 35.3 ± 1.1 (95%C.I.) mg l(-1) Co. Adult zebrafish were exposed (13 d) to sub-lethal (0-25 mg l(-1)) Co and allowed to spawn every 4 d and embryos were collected. After 12-d exposure, fertilisation rate was reduced (6% total eggs fertilised, 25 mg l(-1)) and embryo survival to hatching decreased (60% fertilised eggs survived, 25 mg l(-1)). A concentration-dependent increase in DNA strand breaks was detected in sperm from males exposed (13 d) to Co, and DNA damage in sperm returned to control levels after males recovered for 6 d in clean water. Induction of DNA repair genes (rad51, xrcc5, and xrcc6) in testes was complex and not directly related to Co concentration, although there was significant induction in fish exposed to 15 and 25 mg l(-1) Co relative to controls. Induction of 4.0 ± 0.9, 2.5 ± 0.7, and 3.1 ± 0.7-fold change (mean ± S.E.M. for rad51, xrcc5, and xrcc6, respectively) was observed in testes at the highest Co concentration (25 mg l(-1)). Expression of these genes was not altered in offspring (larvae) spawned after 12-d exposure. Chronic exposure to Co resulted in DNA damage in sperm, induction of DNA repair genes in testes, and indications of reduced reproductive success.

210Po/210Pb dynamics in relation to zooplankton biomass and trophic conditions during an annual cycle in northwestern Mediterranean coastal waters.

Monthly sampling in northwestern Mediterranean coastal waters was undertaken to better understand the relationship between zooplankton biomass and the cycling of the natural radionuclide (210)Po/(210)Pb pair during a one-year period (October 1995-November 1996). In conjunction with mesozooplankton collections and (210)Po/(210)Pb measurements in seawater, zooplankton and their fecal pellets, the biochemical composition of particulate organic matter (POM) was also examined at three depths (0, 20 and 50 m) as an indicator of trophic conditions. During May 1996, a strong zooplankton "bloom" was observed which was preceded by a prolonged increase in POM (protein + carbohydrates + lipids) starting at the end of March, and further demonstrated by a concomitant increase in the concentration of smaller particles, two features that are typical of mesotrophic waters. Simultaneous measurements of (210)Po in sea water and zooplankton showed an inverse trend between these two parameters during the sampling period, with the two lowest (210)Po concentrations in the dissolved phase of seawater coincident with the highest radionuclide concentrations in the zooplankton; however, this apparent relationship was not statistically significant over the entire year. Freshly excreted mesozooplankton and salp fecal pellets, which have been strongly implicated in the removal and downward transport of these radionuclides from the upper water column, contained (210)Po and (210)Pb levels ranging from 175 to 878 and 7.5-486 Bq kg(-1) dry weight, respectively. Salp pellets contained 5 and 10 times more (210)Po and (210)Pb than in fecal pellets produced by mixed zooplankton, a finding most likely related to their different feeding strategies. During the zooplankton biomass peak observed in May, the (210)Po concentration in zooplankton was at a minimum; however, in contrast to what has been reported to occur in some open sea oligotrophic waters, over the year no statistically significant inverse relationship was found between zooplankton biomass and (210)Po concentration in zooplankton. This observation may have resulted from the general lack of very low biomass concentrations (<1 mg m(-3)) measured in these coastal waters, biomass levels which commonly occur in open ocean oligotrophic regions.