Sensitivity of Freshwater Australian Bass (Macquaria novemaculeata) and Silver Perch (Bidyanus bidyanus) to Waterborne Antimony: Exposure-Dose-Response Characteristics and Ion Homeostasis.

We conducted acute toxicity studies using semi-static protocols to examine the lethal responses of Australian bass and silver perch exposed to antimony (Sb) oxidation states in Sb(III) (10.5-30.5 mg L-1) and Sb(V) (95.9-258.7 mg L-1). Bioavailability and the effects of Sb on body ion regulation (Na, Ca, Mg, and K) were also investigated. Antimony species-specific effects were observed with exposure to both Sb oxidation states. Median lethal concentrations (LC50s) for Sb(III) were 13.6 and 18 mg L-1 for Australian bass and silver perch, respectively, and the LC50 for Sb(V) in Australian bass was 165.3 mg L-1. The LC50 could not be calculated for silver perch exposed to Sb(V) as the maximum exposure concentrations produced 40% mortality but a larger-than value of > 258.7 mg L-1 was estimated. Relative median potency values derived from the LC50s were 0.1 Sb(III) and 12.2 and 16.6 Sb(V) for Australian bass and silver perch, respectively, demonstrating greater toxicity of Sb(III) to both fish species. Antimony uptake in fish was observed. Median critical body residue (CBR50) values of 77.7 and 26.6 mg kg-1 for Sb(III) were estimated for Australian bass and silver perch, respectively, and 628.1 mg kg-1 for Sb(V) in Australian bass. Bioconcentration factors (BCFs) for both Sb(III) and Sb(V) did not change with exposure but the greater BCFs for fish exposed to Sb(III) indicate that it is more bioavailable than Sb(V) in acute exposure. No effects on whole-body Na, Ca, Mg, or K ions were observed with fish exposure to either Sb species.

Field and laboratory evaluation of DGT for predicting metal bioaccumulation and toxicity in the freshwater bivalve Hyridella australis exposed to contaminated sediments.

The diffusive gradients in thin films (DGT) technique has shown to be a useful tool for predicting metal bioavailability and toxicity in sediments, however, links between DGT measurements and biological responses have often relied on laboratory-based exposures and further field evaluations are required. In this study, DGT probes were deployed in metal-contaminated (Cd, Pb, Zn) sediments to evaluate relationships between bioaccumulation by the freshwater bivalve Hyridella australis and DGT-metal fluxes under both laboratory and field conditions. The DGT-metal flux measured across the sediment/water interface (±1 cm) was useful for predicting significant cadmium and zinc bioaccumulation, irrespective of the type of sediment and exposure. A greater DGT-Zn flux measured in the field was consistent with significantly higher zinc bioaccumulation, highlighting the importance of performing metal bioavailability assessments in situ. In addition, DGT fluxes were useful for predicting the potential risk of sub-lethal toxicity (i.e., lipid peroxidation and lysosomal membrane damage). Due to its ability to account for multiple metal exposures, DGT better predicted bioaccumulation and toxicity than particulate metal concentrations in sediments. These results provide further evidence supporting the applicability of the DGT technique as a monitoring tool for sediment quality assessment.

Trophic transfer of metals in a seagrass food web: Bioaccumulation of essential and non-essential metals.

Metal concentrations are reported for a seagrass ecosystem receiving industrial inputs. δ13C and δ15N isotope ratios were used to establish trophic links. Copper concentrations (dry mass) ranged from <0.01 µg/g in fish species to 570 µg/g (µâ€¯= 49 ±â€¯SD = 90 µg/g) in the oyster Saccostrea glomerata. Zinc concentrations ranged from 0.6 µg/g in the seagrass Zostera capricorni to 10,800 µg/g in the mud oyster Ostrea angasi (µâ€¯= 434 ±â€¯1390 µg/g). Cadmium concentrations ranged from <0.01 µg/g in fish species to 268 µg/g in Ostrea angasi (µâ€¯= 6 ±â€¯25 µg/g). Lead concentrations ranged from <0.01 µg/g for most fish species to 20 µg/g in polychaetes (µâ€¯= 2 ±â€¯3 µg/g). Biomagnification of metals did not occur. Organisms that fed on particulate organic matter and benthic microalgae had higher metal concentrations than those that fed on detritus. Species physiology also played an important role in the bioaccumulation of metals.

Exposure of the freshwater bivalve Hyridella australis to metal contaminated sediments in the field and laboratory microcosms: metal uptake and effects.

Metal uptake and induced toxic effects on Hyridella australis were investigated by establishing 28 day exposure-dose-response relationships (EDR) of transplanted H. australis at four sites along a sediment metal contamination gradient in the mine affected Molonglo River, NSW. Laboratory exposure of this organism to the same sediments, collected from in situ sites, was run concurrently. Metal concentrations in whole organisms, individual tissues and sub-cellular tissue fractions were measured as organism metal dose. Total antioxidant capacity (TAOC), lipid peroxidation (MDA) and lysosomal membrane destabilisation (LMS) were measured as biological responses. H. australis accumulated significantly higher tissue zinc concentrations compared to the other metals. In situ organisms at the mine affected sites accumulated more metals than organisms in laboratory microcosms. Accumulated zinc, cadmium and the total metal concentrations in whole organism tissues reflected exposure-dose relationships. Sub-cellular analysis showed that most of the accumulated metals, both in the field and laboratory exposed organisms, were detoxified over 28 days exposure. Clear exposure and dose dependent responses of decreased TAOC and measurable increases in MDA and LMS with increased metal exposure and dose were evident in H. australis caged in the river. In contrast, a dose-response relationship was only evident for cadmium in laboratory exposed organisms. Organisms caged at mine affected sites showed stronger EDR relationships than those exposed in laboratory microcosms as they were exposed to additional sources of dissolved zinc and cadmium. Exposure in laboratory microcosms underestimated metal uptake and effects, thus assessment of metal contaminated sediments should be undertaken "in situ".

The response of Isidorella newcombi to copper exposure: Using an integrated biological framework to interpret transcriptomic responses from RNA-seq analysis.

This study describes the transcriptomic response of the Australian endemic freshwater gastropod Isidorella newcombi exposed to 80±1µg/L of copper for 3days. Analysis of copper tissue concentration, lysosomal membrane destabilisation and RNA-seq were conducted. Copper tissue concentrations confirmed that copper was bioaccumulated by the snails. Increased lysosomal membrane destabilisation in the copper-exposed snails indicated that the snails were stressed as a result of the exposure. Both copper tissue concentrations and lysosomal destabilisation were significantly greater in snails exposed to copper. In order to interpret the RNA-seq data from an ecotoxicological perspective an integrated biological response model was developed that grouped transcriptomic responses into those associated with copper transport and storage, survival mechanisms and cell death. A conceptual model of expected transcriptomic changes resulting from the copper exposure was developed as a basis to assess transcriptomic responses. Transcriptomic changes were evident at all the three levels of the integrated biological response model. Despite lacking statistical significance, increased expression of the gene encoding copper transporting ATPase provided an indication of increased internal transport of copper. Increased expression of genes associated with endocytosis are associated with increased transport of copper to the lysosome for storage in a detoxified form. Survival mechanisms included metabolic depression and processes associated with cellular repair and recycling. There was transcriptomic evidence of increased cell death by apoptosis in the copper-exposed organisms. Increased apoptosis is supported by the increase in lysosomal membrane destabilisation in the copper-exposed snails. Transcriptomic changes relating to apoptosis, phagocytosis, protein degradation and the lysosome were evident and these processes can be linked to the degradation of post-apoptotic debris. The study identified contaminant specific transcriptomic markers as well as markers of general stress. From an ecotoxicological perspective, the use of a framework to group transcriptomic responses into those associated with copper transport, survival and cell death assisted with the complex process of interpretation of RNA-seq data. The broad adoption of such a framework in ecotoxicology studies would assist in comparison between studies and the identification of reliable transcriptomic markers of contaminant exposure and response.

The freshwater bivalve Corbicula australis as a sentinel species for metal toxicity assessment: An in situ case study integrating chemical and biomarker analyses.

A weight of evidence approach in environmental assessment includes the use of biomonitor organisms to measure biologically available contaminant concentrations and lethal and sublethal responses in an exposure, dose, and response framework. Corbicula australis was assessed as a test species for metal toxicity using in situ river sediment exposures at 4 locations in the Molonglo River (New South Wales, Australia), which has a legacy of sediment metal contamination, following 8 decades of mining in its upper reaches. A sediment metal contamination gradient was evident from 12.5 km to 47 km downstream of the mine, as follows: zinc (851-130 mg/kg) > lead (104-7 mg/kg) > copper (31-5 mg/kg) > cadmium (2-0.3 mg/kg). Exposed C. australis accumulated the following metals in tissue: zinc (1358-236 µg/g) > copper (24-20 µg/g) > cadmium (4.7-0.7 µg/g) = lead (4.2-1.8 µg/g). Biomarker responses showed increased sublethal impairment with increased tissue metal concentrations. Total antioxidant capacity was mildly impaired, with corresponding increased lipid peroxidation and lysosomal membrane destabilization at the higher tissue metal concentrations. Corbicula australis proved to be an effective biomonitor organism for sediment metal assessment, as it is able to accumulate metals relative to sediment concentrations and showed a pattern of increased sublethal impairment with increased tissue metal concentration. It is recommended as a suitable species for incorporation into local freshwater monitoring and assessment programs. Environ Toxicol Chem 2017;36:709-719. © 2016 SETAC.

The proteasome controls presynaptic differentiation through modulation of an on-site pool of polyubiquitinated conjugates.

Differentiation of the presynaptic terminal is a complex and rapid event that normally occurs in spatially specific axonal regions distant from the soma; thus, it is believed to be dependent on intra-axonal mechanisms. However, the full nature of the local events governing presynaptic assembly remains unknown. Herein, we investigated the involvement of the ubiquitin-proteasome system (UPS), the major degradative pathway, in the local modulation of presynaptic differentiation. We found that proteasome inhibition has a synaptogenic effect on isolated axons. In addition, formation of a stable cluster of synaptic vesicles onto a postsynaptic partner occurs in parallel to an on-site decrease in proteasome degradation. Accumulation of ubiquitinated proteins at nascent sites is a local trigger for presynaptic clustering. Finally, proteasome-related ubiquitin chains (K11 and K48) function as signals for the assembly of presynaptic terminals. Collectively, we propose a new axon-intrinsic mechanism for presynaptic assembly through local UPS inhibition. Subsequent on-site accumulation of proteins in their polyubiquitinated state triggers formation of presynapses.

Sediment Metal Concentration Survey Along the Mine-Affected Molonglo River, NSW, Australia.

Metal concentrations were measured in sediments of the mine-affected Molonglo River to determine current metal concentrations and distribution along the river. Compared with an uncontaminated site at 6.5 km upstream of the Captains Flat mine, sediments collected from the river at ≤12.5 km distance below the mine had a significantly higher percentage of finely divided silt and clay with higher concentrations of cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn). The measured metal concentrations in the mine affected sites of the river were in the following order: Zn = 697-6818 > Pb = 23-1796 > Cu = 10-628 > Cd = 0.13-8.7 µg/g dry mass. The highest recorded metal concentrations were Cd at 48, Cu at 45, Pb at 240, and Zn at 81 times higher than the background concentrations of these metals in the river sediments. A clear sediment metal-contamination gradient from the mine site to 63 km downstream was established for Cd, Cu, Pb, and Zn in the river sediments. Compared with sediment metal concentrations before a major flood in 2010, only Zn concentrations increased. For all of the mine-affected sites studied, Cd and Zn concentrations exceeded the (ANZECC/ARMCANZ, Australian and New Zealand guidelines for fresh and marine water quality. Australian and New Zealand Environment and Conservation Council/Agriculture and Resource Management Council of Australia and New Zealand, 2000) interim sediment-quality guidelines low values for Cd (1.5 µg/g dry mass) and the high value for Zn (410 µg/g dry mass). Existing metal loads in the riverbed sediments may still be adversely affecting the river infauna.

Effects of cadmium accumulation from suspended sediments and phytoplankton on the Oyster Saccostrea glomerata.

Metals are accumulated by filter feeding organisms via water, ingestion of suspended sediments or food. The uptake pathway can affect metal toxicity. Saccostrea glomerata were exposed to cadmium through cadmium-spiked suspended sediments (19 and 93µg/g dry mass) and cadmium-enriched phytoplankton (1.6-3µg/g dry mass) and cadmium uptake and effects measured. Oysters accumulated appreciable amounts of cadmium from both low and high cadmium spiked suspended sediment treatments (5.9±0.4µg/g and 23±2µg/g respectively compared to controls 0.97±0.05µg/g dry mass). Only a small amount of cadmium was accumulated by ingestion of cadmium-enriched phytoplankton (1.9±0.1µg/g compared to controls 1.2±0.1µg/g). In the cadmium spiked suspended sediment experiments, most cadmium was desorbed from sediments and cadmium concentrations in S. glomerata were significantly related to dissolved cadmium concentrations (4-21µg/L) in the overlying water. In the phytoplankton feeding experiment cadmium concentrations in overlying water were <0.01µg/L. In both exposure experiments, cadmium-exposed oysters showed a significant reduction in total antioxidant capacity and significantly increased lipid peroxidation and percentage of destabilised lysosomes. Destabilised lysosomes in the suspended sediments experiments also resulted from stress of exposure to the suspended sediments. The study demonstrated that exposure to cadmium via suspended sediments and to low concentrations of cadmium through the ingestion of phytoplankton, can cause sublethal stress to S. glomerata.

Importance of subcellular metal partitioning and kinetics to predicting sublethal effects of copper in two deposit-feeding organisms.

The role of subcellular partitioning of copper on the sublethal effects to two deposit-feeding organisms (41-day growth in the bivalve Tellina deltoidalis and 11-day reproduction in the amphipod Melita plumulosa) was assessed for copper-spiked sediments with different geochemical properties. Large differences in bioaccumulation and detoxification strategies were observed. The bivalve accumulated copper faster than the amphipod, and can be considered a relatively strong net bioaccumulator. The bivalve, however, appears to regulate the metabolically available fraction (MAF) of the total metal pool by increasing the net accumulation rate of copper in the biologically detoxified metal pool (BDM), where most of the copper is stored. In the amphipod, BDM concentration remained constant with increasing copper exposures and it can be considered a very weak net bioaccumulator of copper. This regulation of copper, with relatively little stored in detoxified forms, appears to best describe the strategy applied by the amphipod to minimize the potential toxic effects of copper. When the EC50 values for growth and reproduction are expressed based on the MAF of copper, the sensitivity of the two species appears similar, however when expressed based on the net accumulation rate of copper in the metabolically available fraction (MAFrate), the bivalve appears more sensitive to copper. These results indicate that describing the causality of metal effects in terms of kinetics of uptake, detoxification, and excretion rather than threshold metal body concentrations is more effective in predicting the toxic effects of copper. Although the expression of metal toxicity in terms of the rate at which the metal is bioaccumulated into metabolically available forms may not be feasible for routine assessments, a deeper understanding of uptake rates from all exposure routes may improve our ability to assess the risk posed by metal-contaminated sediments.

Modeling food web structure and selenium biomagnification in Lake Macquarie, New South Wales, Australia, using stable carbon and nitrogen isotopes.

As a consequence of coal-fired power station operations, elevated selenium concentrations have been reported in the sediments and biota of Lake Macquarie (New South Wales, Australia). In the present study, an ecosystem-scale model has been applied to determine how selenium in a seagrass food web is processed from sediments and water through diet to predators, using stable isotopes (δ(13) C and δ(15) N) to establish the trophic position of organisms. Trophic position, habitat, and feeding zone were examined as possible factors influencing selenium bioaccumulation. Selenium concentrations ranged from 0.2 µg/g dry weight in macroalgae species to 12.9 µg/g in the carnivorous fish Gerres subfasciatus. A mean magnification factor of 1.39 per trophic level showed that selenium is biomagnifying in the seagrass food web. Habitat and feeding zone influenced selenium concentrations in invertebrates, whereas feeding zone was the only significant factor influencing selenium concentrations in fish. The sediment-water partitioning coefficient (Kd ) of 4180 showed that partitioning of selenium entering the lake to particulate organic material (POM) is occurring, and consequently availability to food webs from POM is high. Trophic transfer factors (invertebrate = 1.9; fish = 1.2) were similar to those reported for other water bodies, showing that input source is not the main determinant of the magnitude of selenium bioaccumulation in a food web, but rather the initial partitioning of selenium into bioavailable POM. Environ Toxicol Chem 2015;34:608-617. © 2014 SETAC.

Bioavailability and toxicity of zinc from contaminated freshwater sediments: linking exposure-dose-response relationships of the freshwater bivalve Hyridella australis to zinc-spiked sediments.

To evaluate the use of the freshwater bivalve Hyridella australis as a potential biomonitor for zinc contamination in freshwater sediments, the bioavailability and toxicity of zinc contaminated sediments (low 44 ± 5, medium 526 ± 41, high 961 ± 38 µg/g dry mass) were investigated in laboratory microcosms for 28 days by examining H. australis exposure-dose-response relationships. Zinc concentrations in sediments and surface waters were measured as zinc exposure. Zinc in whole organism soft body tissues and five individual tissues were measured as organism zinc dose. Sub-cellular localisation of zinc in hepatopancreas tissues was investigated to further understand the zinc handling strategies and tolerance of H. australis. Total antioxidant capacity, lipid peroxidation and lysosomal membrane stability were measured in hepatopancreas tissues as zinc induced biomarker responses. Accumulated zinc concentrations in whole body tissues of H. australis reflected the zinc exposure and exhibited exposure dependent zinc accumulation at day 28. Gills accumulated significantly higher zinc concentrations than other tissues, however, no significant differences in zinc accumulation between treatments were detected for any of the individual tissues analysed. Analysis of individual tissue zinc concentrations, therefore, may not offer any advantages for monitoring bioavailable zinc in freshwater environments with this organism. Relationships between tissue zinc and calcium concentration suggest accumulation of zinc by H. australis may have occurred as an analogue of calcium which is a major constituent in shell and granules of unionid bivalves. A high percentage of accumulated zinc in the hepatopancreas tissues was detoxified and stored in metallothionein like proteins and metal rich granules. Of the zinc accumulated in the biologically active metal pool, 59-70% was stored in the lysosome+microsome fraction. At the concentrations tested, increasing zinc exposure resulted in decreasing total antioxidant capacity and measurable increases in the sublethal effects, lipid peroxidation and lysosomal membrane destabilisation, were observed. Based on exposure-dose analysis, H. australis partially regulates zinc uptake and weakly exhibits bioavailability of zinc in freshwater environments, however, exposure-response analysis shows zinc induced toxicological effects, suggesting the potential of this organism as a biomonitor for zinc in heavily contaminated freshwater environments.

Exposure-dose-response of Tellina deltoidalis to contaminated estuarine sediments 3. Selenium spiked sediments.

The metalloid selenium is an essential element which at slightly elevated concentrations is toxic and mutagenic. In Australia the burning of coal for power generation releases selenium into estuarine environments where it accumulates in sediments. The relationship between selenium exposure, dose and response was investigated in the deposit feeding, benthic, marine bivalve Tellina deltoidalis. Bivalves were exposed in microcosms for 28 days to individual selenium spiked sediments, 0, 5 and 20 µg/g dry mass. T. deltoidalis accumulated selenium from spiked sediment but not in proportion to the sediment selenium concentrations. The majority of recovered subcellular selenium was associated with the nuclei and cellular debris fraction, probably as protein bound selenium associated with plasma and selenium bound directly to cell walls. Selenium exposed organisms had increased biologically detoxified selenium burdens which were associated with both granule and metallothionein like protein fractions, indicating selenium detoxification. Half of the biologically active selenium was associated with the mitochondrial fraction with up to 4 fold increases in selenium in exposed organisms. Selenium exposed T. deltoidalis had significantly reduced GSH:GSSG ratios indicating a build-up of oxidised glutathione. Total antioxidant capacity of selenium exposed T. deltoidalis was significantly reduced which corresponded with increased lipid peroxidation, lysosomal destabilisation and micronuclei frequency. Clear exposure-dose-response relationships have been demonstrated for T. deltoidalis exposed to selenium spiked sediments, supporting its suitability for use in selenium toxicity tests using sub-lethal endpoints.

Exposure-dose-response relationships of the freshwater bivalve Hyridella australis to cadmium spiked sediments.

To understand how benthic biota may respond to the additive or antagonistic effects of metal mixtures in the environment it is first necessary to examine their responses to the individual metals. In this context, laboratory controlled single metal-spiked sediment toxicity tests are useful to assess this. The exposure-dose-response relationships of Hyridella australis to cadmium-spiked sediments were, therefore, investigated in laboratory microcosms. H. australis was exposed to individual cadmium spiked sediments (<0.05 (control), 4±0.3 (low) and 15±1 (high) µg/g dry mass) for 28 days. Dose was measured as cadmium accumulation in whole soft body and individual tissues at weekly intervals over the exposure period. Dose was further examined as sub-cellular localisation of cadmium in hepatopancreas tissues. The biological responses in terms of enzymatic and cellular biomarkers were measured in hepatopancreas tissues at day 28. H. australis accumulated cadmium from spiked sediments with an 8-fold (low exposure organisms) and 16-fold (high exposure organisms) increase at day 28 compared to control organisms. The accumulated tissue cadmium concentrations reflected the sediment cadmium exposure at day 28. Cadmium accumulation in high exposure organisms was inversely related to the tissue calcium concentrations. Gills of H. australis showed significantly higher cadmium accumulation than the other tissues. Accumulated cadmium in biologically active and biologically detoxified metal pools was not significantly different in cadmium exposed organisms, which suggests that H. australis has some tolerance to cadmium. The metallothionein like protein fraction played an important role in the sequestration and detoxification of cadmium and the amount sequestered in this fraction increased with increased cadmium exposure. The highest percentage of biologically active cadmium was associated with the lysosome+microsome and mitochondrial fractions. Cadmium concentrations in these two fractions of cadmium exposed organisms were significantly higher with respect to controls. Total antioxidant capacity decreased with increased cadmium exposure and tissue dose. Lipid peroxidation increased and lysosomal membrane stability decreased significantly with increased cadmium exposure and tissue dose. Based on exposure-dose-response analysis in this study, H. australis would be a suitable organism for assessing cadmium sediment exposure and toxicity.

Effects of lead-spiked sediments on freshwater bivalve, Hyridella australis: linking organism metal exposure-dose-response.

Lead entering aquatic ecosystems adsorbs to sediments and has the potential to cause adverse effects on the health of benthic organisms. To evaluate the freshwater bivalve Hyridella australis as a bioindicator for sediment toxicity, their exposure-dose and response to lead contaminated sediments (< 0.01, 205 ± 9 and 419 ± 16 µg/g dry mass) was investigated in laboratory microcosms using 28 day exposures. Despite high concentrations of lead in the sediments, organisms accumulated low concentrations of lead in their tissues after 28 days of exposure (low treatment: 2.2 ± 0.2 µg/g dry mass, high treatment: 4.2 ± 0.1 µg/g dry mass), however, accumulated lead concentrations in lead exposed organisms were two fold (low treatment) and four fold (high treatment) higher than that of unexposed organisms (1.2 ± 0.3 µg/g dry mass). Accumulation of lead by H. australis may have occurred as analogues of calcium and magnesium. Labial palps accumulated significantly more lead than other tissues. Of the lead accumulated in the hepatopancreas, 83%-91% was detoxified and stored in metal rich granules. The proportions and concentrations of lead in this fraction increased with lead exposure, which suggests that lead detoxification pathway plays an important role in metal tolerance of H. australis. The biologically active lead was mainly present in the mitochondrial fraction which increased with lead exposure. Total antioxidant capacity of H. australis significantly decreased while lipid peroxidation and lysosomal membrane destabilation increased with lead exposure. This study showed a clear lead exposure-dose-response relationship and indicates that H. australis would be a good biomonitor for lead in freshwater ecosystems.

Exposure-dose-response of Tellina deltoidalis to metal contaminated estuarine sediments 2. Lead spiked sediments.

Lead accumulation in estuarine sediments, as a result of activities such as mining and ore smelting, and through urban runoff is a continuing problem in the increasingly developed world. Marine organisms accumulate lead, which is known to be highly toxic to biological processes and to degrade organism and ecosystem health. Here the relationship between lead exposure, dose and response was investigated in the sediment dwelling, deposit feeding, marine bivalve Tellina deltoidalis. Bivalves were exposed in the laboratory to individual lead spiked sediments at < 0.01, 100 and 300 µg/g dry mass, for 28 days and accumulated total tissue lead concentrations of 4, 96 and 430 µg/g, respectively. Subcellular fractionation indicated that around 70% of the total accumulated tissue lead was detoxified, three quarters of the detoxified lead fraction was converted into metal rich granules, with the remainder in the metallothionein like protein fraction. The majority of biologically active lead was associated with the mitochondrial fraction with up to a 128 fold increase in lead burden in exposed organisms compared to controls. This indicates lead detoxification was occurring but the organism was unable to prevent lead interacting with sensitive organelles. With increased lead exposure T. deltoidalis showed a suppression in glutathione peroxidase activity, total glutathione concentration and reduced GSH:GSSG ratios, however, these differences were not significant. Lead exposed T. deltoidalis had a significantly reduced total antioxidant capacity which corresponded with increased lipid peroxidation, lysosomal destabilisation and micronuclei frequency. The exposure-dose-response relationships demonstrated for lead exposed T. deltoidalis supports its potential for the development of sublethal endpoints in lead toxicity assessment.

Exposure-dose-response of Tellina deltoidalis to metal-contaminated estuarine sediments: 1. Cadmium spiked sediments.

Cadmium is a ubiquitous environmental metal contaminant with an affinity for biological membranes; it can enter cells by facilitated transport and it binds therein to various biomolecules and affects membrane system function. The relationship between cadmium exposure, dose and response was investigated in the benthic, deposit feeding, marine bivalve Tellina deltoidalis, using 28 day microcosm spiked cadmium exposures. Tissue cadmium reached steady state with the exposure concentration. Half the accumulated cadmium was detoxified and with increased exposure more was converted into metal rich granules. Most biologically active cadmium was in the mitochondrial fraction, with up to 7320-fold cadmium increases in exposed organisms. Cadmium exposed T. deltoidalis generally had reduced glutathione peroxidase enzyme activity. An increase in total glutathione concentrations, due to a build up of oxidised glutathione, was indicated by the reduced to oxidised glutathione ratio. All cadmium exposed T. deltoidalis had reduced total antioxidant capacity that corresponded with increased lipid peroxidation, lysosomal destabilisation and micronuclei frequency. Clear exposure-dose-response relationships have been demonstrated for T. deltoidalis exposed to cadmium-spiked sediments, supporting this organism's suitability for laboratory or in situ evaluation of sediment cadmium toxicity.

Exposure-dose-response of Anadara trapezia to metal contaminated estuarine sediments: 3. Selenium spiked sediments.

Selenium enters near shore marine environments from the activities of coal-fired power stations. Although selenium is an essential element, at elevated concentrations it can cause genotoxic damage. The relationship between selenium exposure dose and response was investigated in Anadara trapezia exposed to selenium spiked sediment (5 µg/g and 20 µg/g dry mass) for 56 days. A. trapezia reached an equilibrium selenium tissue concentration (2 µg/g and 10 µg/g respectively) by day 42. Gills had significantly more selenium than the hepatopancreas and haemolymph. Between 12 and 21% of accumulated selenium in the gill and hepatopancreas was detoxified and in the metal rich granule. Most of the biologically active selenium in both tissues was in the mitochondrial fraction. Glutathione peroxidase activity and mean total glutathione concentrations for selenium exposed organisms were not significantly different to controls. The ratio of reduced to oxidised glutathione and the total antioxidant capacity were significantly reduced in selenium exposed organisms compared to control organisms. Increased selenium exposure resulted in significant increases in lipid peroxidation, lysosomal destabilisation and an increased frequency of micronuclei. A significant exposure-dose-response relationship for A. trapezia exposed to selenium enriched sediments indicates that elevated sediment selenium concentrations can increased biologically active selenium burdens and cause impairment of cellular processes and cell integrity.

Exposure-dose-response of Anadara trapezia to metal contaminated estuarine sediments. 2. Lead spiked sediments.

The composition of near shore marine environments is increasingly being altered by contaminants from human activities. The ability of lead, which has no known biological function, to mimic biologically essential metals makes it one of the most toxic to marine biota. The relationship between lead exposure, dose and response was investigated in Anadara trapezia exposed for 56 days to lead spiked sediment (100 and 300 µg/g dry mass). Lead tissue concentrations of the 300 µg/g exposed A. trapezia doubled in the last 2 weeks of exposure with final lead tissue concentrations of exposed organisms of 1 and 12 µg/g, respectively. Tissue lead accumulation of exposed organisms followed the pattern haemolymph>gill>hepatopancreas during much of the 56 day exposure. Between 30 and 69% of accumulated lead in the gill and hepatopancreas was detoxified and fairly evenly distributed between the metal rich granule and the metallothionein like protein fractions. Approximately half of the biologically active lead in both tissues was in the mitochondrial fraction which showed increased cytochrome c oxidase activity in lead exposed organisms. There was a reduction in GPx activity, an associated increase in total glutathione concentrations and reduced GSH:GSSG ratios due to a build up of oxidised glutathione. These changes in the glutathione pathway were reflected in the total antioxidant capacity of lead exposed A. trapezia which were significantly reduced compared to control organisms. Increased lead exposure significantly increased lipid peroxidation, lysosomal destabilisation and frequency of micronuclei. A significant exposure-dose-response relationship for A. trapezia exposed to lead enriched sediments indicates that elevated sediment lead concentrations have the potential to increase biologically active lead burdens and impair the antioxidant reduction capacity leading to a series of associated effects from lipid peroxidation to cellular perturbation and genotoxic damage.