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.

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".

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.

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.

Postsynaptic decoding of neural activity: eEF2 as a biochemical sensor coupling miniature synaptic transmission to local protein synthesis.

Activity-dependent regulation of dendritic protein synthesis is critical for enduring changes in synaptic function, but how the unique features of distinct activity patterns are decoded by the dendritic translation machinery remains poorly understood. Here, we identify eukaryotic elongation factor-2 (eEF2), which catalyzes ribosomal translocation during protein synthesis, as a biochemical sensor in dendrites that is specifically and locally tuned to the quality of neurotransmission. We show that intrinsic action potential (AP)-mediated network activity in cultured hippocampal neurons maintains eEF2 in a relatively dephosphorylated (active) state, whereas spontaneous neurotransmitter release (i.e., miniature neurotransmission) strongly promotes the phosphorylation (and inactivation) of eEF2. The regulation of eEF2 phosphorylation is responsive to bidirectional changes in miniature neurotransmission and is controlled locally in dendrites. Finally, direct spatially controlled inhibition of eEF2 phosphorylation induces local translational activation, suggesting that eEF2 is a biochemical sensor that couples miniature synaptic events to local translational suppression in neuronal dendrites.

Microfluidic and compartmentalized platforms for neurobiological research.

Methods to compartmentalize neurons allow distinct neuronal segments (i.e., cell bodies, axons, dendrites, or synapses) to be accessed, visualized, and/or manipulated. Compartmentalization has resulted in multiple studies that would not otherwise be possible in vivo or in traditional random cultures, such as investigations of axonal transport, biochemical analysis of axons, and axonal injury/regeneration. Chambers for compartmentalizing neurons were first developed for long projection peripheral neurons in the 1970s using machined Teflon dividers and relied on manually applied grease layers to spatially and fluidically separate distal axons from their cell bodies. More recently microfabrication and soft lithography techniques have been used to create compartmentalized microfluidic platforms, relying on microgrooves contained within a solid barrier through which axons and dendrites are able to extend, but not their cell bodies. These platforms are unique in their ability to culture central nervous system (CNS) neurons and allow high-resolution live imaging. These microfluidic platforms have allowed new investigations of axonal and synaptic biology in the CNS. Moreover, these microfluidic platforms offer improvements for other neural cell and tissue preparations. In this review we discuss traditional methods for compartmentalization, compartmentalized microfluidic platforms, and their use for neurobiology. Lastly, we discuss the use of these platforms for defining and manipulating synapses both pharmacologically and by electrical stimulation and recording.

Axonal mRNA in uninjured and regenerating cortical mammalian axons.

Using a novel microfluidic chamber that allows the isolation of axons without contamination by nonaxonal material, we have for the first time purified mRNA from naive, matured CNS axons, and identified the presence of >300 mRNA transcripts. We demonstrate that the transcripts are axonal in nature, and that many of the transcripts present in uninjured CNS axons overlap with those previously identified in PNS injury-conditioned DRG axons. The axonal transcripts detected in matured cortical axons are enriched for protein translational machinery, transport, cytoskeletal components, and mitochondrial maintenance. We next investigated how the axonal mRNA pool changes after axotomy, revealing that numerous gene transcripts related to intracellular transport, mitochondria and the cytoskeleton show decreased localization 2 d after injury. In contrast, gene transcripts related to axonal targeting and synaptic function show increased localization in regenerating cortical axons, suggesting that there is an increased capacity for axonal outgrowth and targeting, and increased support for synapse formation and presynaptic function in regenerating CNS axons after injury. Our data demonstrate that CNS axons contain many mRNA species of diverse functions, and suggest that, like invertebrate and PNS axons, CNS axons synthesize proteins locally, maintaining a degree of autonomy from the cell body.

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.

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.

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 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.

Microfluidic chambers for cell migration and neuroscience research.

This chapter describes the fabrication and use microfluidic chambers for cell migration and neuroscience research. Both microfluidic chambers are made using soft lithography and replica molding. The main advantages of using soft lithography to create microfluidic chambers are reproducibility, ease of use, and straightforward fabrication procedures. The devices can be fabricated in biology and chemistry laboratories with minimal access to clean-room facilities. First, a microfluidic chemotaxis chamber, which has been used in investigating chemotaxis of neutrophils, human breast cancer cells, and other cell types, is described. Precise and stable gradients of chemoattractants with arbitrary shapes can be generated for different applications. Second, a multicompartment culture chamber that can fluidically isolate neuronal processes from cell bodies is described. The design of this chamber is such that only neurites grow through a series of microgrooves embedded in a physical barrier. Both devices are compatible with phase, differential interference contrast, and fluorescence microscopy.

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.

Patterned cell culture inside microfluidic devices.

This paper describes a simple plasma-based dry etching method that enables patterned cell culture inside microfluidic devices by allowing patterning, fluidic bonding and sterilization steps to be carried out in a single step. This plasma-based dry etching method was used to pattern cell-adhesive and non-adhesive areas on the glass and polystyrene substrates. The patterned substrate was used for selective attachment and growth of human umbilical vein endothelial cells, MDA-MB-231 human breast cancer cells, NIH 3T3 mouse fibroblasts, and primary rat cortical neurons. Finally, we have successfully combined the dry-patterned substrate with a microfluidic device. Patterned primary rat neurons were maintained for up to 6 days inside the microfluidic devices and the neurons' somas and processes were confined to the cell-adhesive region. The method developed in this work offers a convenient way of micropatterning biomaterials for selective attachment of cells on the substrates, and enables culturing of patterned cells inside microfluidic devices for a number of biological research applications where cells need to be exposed to well-controlled fluidic microenvironment.

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.

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.

Microfluidic Multicompartment Device for Neuroscience Research.

This paper describes and characterizes a novel microfabricated neuronal culture device. This device combines microfabrication, microfluidic, and surface micropatterning techniques to create a multicompartment neuronal culturing device that can be used in a number of neuroscience research applications. The device is fabricated in poly(dimethylsiloxane), PDMS, using soft lithography techniques. The PDMS device is placed on a tissue culture dish (polystyrene) or glass substrate, forming two compartments with volumes of less than 2 µL each. These two compartments are separated by a physical barrier in which a number of micron-size grooves are embedded to allow growth of neurites across the compartments while maintaining fluidic isolation. Cells are plated into the somal (cell body) compartment, and after 3-4 days, neurites extend into the neuritic compartment via the grooves. Viability of the neurons in the devices is between 50 and 70% after 7 days in culture; this is slightly lower than but comparable to values for a control grown on tissue culture dishes. Healthy neuron morphology is evident in both the devices and controls. We demonstrate the ability to use hydrostatic pressure to isolate insults to one compartment and, thus, expose localized areas of neurons to insults applied in soluble form. Due to the high resistance of the microgrooves for fluid transport, insults are contained in the neuritic compartment without appreciable leakage into the somal compartment for over 15 h. Finally, we demonstrate the use of polylysine patterning in combination with the microfabricated device to facilitate identification and visualization of neurons. The ability to direct sites of neuronal attachment and orientation of neurite outgrowth by micropatterning techniques, combined with fluidically isolated compartments within the culture area, offers significant advantages over standard open culture methods and other conventional methods for manipulating distinct neuronal microenvironments.

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 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.

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.