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.

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".

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 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.