Bioavailability and Chronic Toxicity of Metal Sulfide Minerals to Benthic Marine Invertebrates: Implications for Deep Sea Exploration, Mining and Tailings Disposal.

The exploration and proposed mining of sulfide massive deposits in deep-sea environments and increased use deep-sea tailings placement (DSTP) in coastal zones has highlighted the need to better understand the fate and effects of mine-derived materials in marine environments. Metal sulfide ores contain high concentrations of metal(loid)s, of which a large portion exist in highly mineralized or sulfidised forms and are predicted to exhibit low bioavailability. In this study, sediments were spiked with a range of natural sulfide minerals (including chalcopyrite, chalcocite, galena, sphalerite) to assess the bioavailability and toxicity to benthic invertebrates (bivalve survival and amphipod survival and reproduction). The metal sulfide phases were considerably less bioavailable than metal contaminants introduced to sediment in dissolved forms, or in urban estuarine sediments contaminated with mixtures of metal(loid)s. Compared to total concentrations, the dilute-acid extractable metal(loid) (AEM) concentrations, which are intended to represent the more oxidized and labile forms, were more effective for predicting the toxicity of the sulfide mineral contaminated sediments. The study indicates that sediment quality guidelines based on AEM concentrations provide a useful tool for assessing and monitoring the risk posed by sediments impacted by mine-derived materials in marine environments.

Assessing the Effects of Bioturbation on Metal Bioavailability in Contaminated Sediments by Diffusive Gradients in Thin Films (DGT).

The burrowing and feeding activities of benthic organisms can alter metal speciation in sediments and affect an organisms' exposure to metals. Recently, the performance of the in situ technique of diffusive gradients in thin films (DGT) for predicting metal bioavailability has been investigated in response to the increasing demand of considering contaminant bioavailability in sediment quality assessments. In this study, we test the ability of the DGT technique for predicting the metal bioavailability in clean and contaminated sediments that are being subjected to varying degrees of sediments disturbance: low bioturbation (bivalve Tellina deltoidalis alone) and high bioturbation (bivalve and actively burrowing amphipod, Victoriopisa australiensis). Significant release of DGT-labile Cd, Ni, Pb, and Zn, but lower Cu and Fe, occurred in the pore and overlying waters of sediments exposed to high bioturbation conditions, resulting in higher bioaccumulation of zinc in bivalves. Strong relationships were found between bioaccumulation of Pb and Zn and time-integrated DGT-metal fluxes, whereas poor relationships were obtained using total or dilute-acid extractable metal concentrations. This results demonstrate that DGT is a useful tool for assessing metal bioavailability in sediments and can provide useful predictions of metal bioavailable to benthic organisms in dynamic sediment environments.

Dissolved and particulate copper exposure induces differing gene expression profiles and mechanisms of toxicity in the deposit feeding amphipod Melita plumulosa.

Uptake of metals via ingestion is an important route of exposure for many invertebrates, and it has been suggested that the toxic response to metals accumulated via food differs from that of metals accumulated via the dissolved phase. To test this hypothesis, the deposit-feeding epibenthic amphipod Melita plumulosa was exposed to nontoxic or reproductively toxic concentrations of copper via the overlying water, via ingestion of sediment, or via a combination of the two. Rates of copper uptake from the two exposure routes were predicted using a biokinetic model. Gene expression profiles were measured via microarray analysis and confirmed via quantitative polymerase chain reaction. Differences in expression profiles were related to the exposure route more than to individual or combined rates of copper uptake. Chitinase and digestive protease transcript expression levels correlated to the copper uptake rate from sediment, rather than from the dissolved phase or combined total uptake rate. Overall, this study supports the hypothesis that metals accumulated via ingestion have a different mode of toxic action than metals taken up from water. Consequently, guidelines that only consider dissolved metal exposure, including equilibrium-partitioning-based guidelines, may underestimate the potential effects from deposited or resuspended metal-contaminated sediments.

Sub-lethal effects of copper to benthic invertebrates explained by sediment properties and dietary exposure.

The next generation of sediment quality guidelines (SQGs) requires better established causal links between the chronic exposure and effects of metals from both dissolved and dietary sources. The potential for dietary exposure from sediment metals to cause toxic effects to benthic invertebrates is strongly influenced by the metal-binding properties of the sediments. For relatively oxidized sediments, sublethal effects of copper to the epibenthic deposit-feeding amphipod, Melita plumulosa, and the benthic harpacticoid copepod, Nitocra spinipes, were investigated. Effects on reproduction were strongly influenced by the properties of the sediments and sediment-bound copper was found to be the major contribution to the toxicity. For sediments with the same total copper concentrations, effects were less for sediments with greater concentrations of fine particles (<63 µm sediment) or particulate organic carbon (OC). The OC-normalized copper concentration in the <63 µm sediment fraction provided a single effects threshold for all sediment types. For M. plumulosa and N. spinipes, the 10% effect concentrations (EC10s) were 5.2 and 4.8 mg <63 µm Cu g(-1) OC. These chronic EC10s indicate that a SQG of 3.5 mg <63 µm Cu g(-1) OC, that was previously proposed based on a species sensitivity distribution of acute no effects thresholds data for 12 benthic organisms, will be protective for these species. The study confirms the appropriateness of using SQGs that vary with sediment properties and that SQGs of this form provide adequate protection for metal exposure via both dissolved and dietary exposure pathways.

The influence of salinity on the chronic toxicity of shale gas flowback wastewater to freshwater organisms.

The potential environmental risk associated with flowback waters generated during hydraulic fracturing of target shale gas formations needs to be assessed to enable management decisions and actions that prevent adverse impacts on aquatic ecosystems. Using direct toxicity assessment (DTA), we determined that the shale gas flowback wastewater (FWW) from two exploration wells (Tanumbirini-1 and Kyalla 117 N2) in the Beetaloo Sub-basin, Northern Territory, Australia were chronically toxic to eight freshwater biota. Salinity in the respective FWWs contributed 16% and 55% of the chronic toxicity at the 50% effect level. The remaining toxicity was attributed to unidentified chemicals and interactive effects from the mixture of identified organics, inorganics and radionuclides. The most sensitive chronic endpoints were the snail (Physa acuta) embryo development (0.08-1.1% EC10), microalga (Chlorella sp. 12) growth rate inhibition (0.23-3.7% EC10) and water flea (Ceriodaphnia cf. dubia) reproduction (0.38-4.9% EC10). No effect and 10% effect concentrations from the DTA were used in a species sensitivity distribution to derive "safe" dilutions of 1 in 300 and 1 in 1140 for the two FWWs. These dilutions would provide site-specific long-term protection to 95% of aquatic biota in the unlikely event of an accidental spill or seepage.

Probabilistic risk assessment of mine-derived copper in the Ok Tedi/Fly River, Papua New Guinea.

The Ok Tedi mine discharges waste rock and tailings into the Ok Tedi River in Papua New Guinea. This has resulted in elevated copper concentrations throughout the Ok Tedi/Fly River system, which can potentially impact aquatic biota. Ten years of measured copper and toxicity monitoring data were used to assess the risk of chronic effects from the mine-derived copper. Cumulative probability plots of dissolved and labile copper were compared to a species sensitivity distribution (SSD) of published copper toxicity data for four regions of the river. The Cu-SSD was used to estimate the risk of chronic effects to aquatic organisms in the Ok Tedi/Fly River at a range of potential copper exposure scenarios. The risk to species at the median labile copper concentration for each region showed a gradient effect with distance downstream from the mine and only the most sensitive (0.2-11%) species were at risk. There were copper exceedances of the region-specific guideline values (GV) and default guideline value (DGV) 88% and 74% of the time, respectively, in the Ok Tedi region (closest to the mine) and this is considered a high risk of chronic effects. Measured copper concentrations in the middle Fly River, lower Fly River (farthest downstream of the mine) and the river at Kiunga (reference site) exceeded the region-specific GVs and DGVs less frequently to rarely and present a lower risk of chronic effects from copper. The risk was supported using toxicity tests with the local microalgal species Chlorella sp. Comparison of recent (2010-2020) and historical (1996-2004) copper monitoring data from the Ok Tedi/Fly River indicates a decrease in the labile copper concentrations (30-76%) at key sites from impacted regions and a subsequent decrease in risk. This coincides with improved mining practices aimed at reducing the copper load into the Ok Tedi/Fly River.

An assessment of three harpacticoid copepod species for use in ecotoxicological testing.

The relatively short life cycles of harpacticoid copepods makes them appropriate animals for use in tests that rapidly assess the acute, sublethal, or chronic effects of sediment contaminants. In this study, four harpacticoid copepod species (Nitocra spinipes, Tisbe tenuimana, Robertgurneya hopkinsi, and Halectinosoma sp.) were isolated from clean marine sediments, and procedures for laboratory culturing were developed. Halectinosoma sp. was abandoned due to handling difficulties. For the remaining species, the influence of food type and quantity on life-cycle progression was assessed. A mixed diet, comprising two species of algae (Tetraselmis sp. and Isochrysis sp.) and fish food (Sera Micron) was found to maintain healthy cultures and was fed during laboratory tests. Water-only exposure to dissolved copper (Cu) showed that the times (range) required to cause 50% lethality (LT(50)) were 24 (22-27) h at 50 µg Cu/l for T. tenuimana; 114 (100-131) and 36 (32-40) h for 200 and 400 µg Cu/l, respectively, for N. spinipes; and 119 (71-201) and 25 (18-33) h for 200 and 400 µg Cu/l, respectively, for R. hopkinsi. 96-h LC(50) (concentration causing 50% lethality) were also determined for adult N. spinipes exposed to cadmium, copper, zinc, ammonia, and phenol. A ranking system was generated based on the ease handling and culturing, rate of maturity, food selectivity and sensitivity to Cu. From this ranking, N. spinipes was determined to be the most suitable species for use in developing sediment-toxicity tests. The measurement of total reproductive output of N. spinipes during 10-day exposure to whole sediment was found to provide a useful end point for assessing the effects of sediment contamination.

Improved prediction of sediment toxicity using a combination of sediment and overlying water contaminant exposures.

The choice of sediment quality assessment methodologies can strongly influence assessment outcomes and management decisions for contaminated sites. While in situ (field) methods may potentially provide greater realism, high costs and/or complex logistics often prevent their use and assessment must rely on laboratory-based methods. In this study, we utilised static-renewal and flow-through ecotoxicology tests in parallel on sediments with a wide range of properties and varying types and concentrations of contaminants. The prediction of chronic effects to amphipod reproduction was explored using multiple linear regression (MLR). The study confirmed the considerable over-estimation of the risk of toxicity of contaminated sediments in field locations when assessments rely on the results of laboratory-based static and static-renewal tests. Improved prediction of toxicity risks was achieved using a combination of contaminant exposure measures from sediment and overlying water. Existing sediment and water quality guideline values (GVs) were effective for predicting risks posed by sediments containing mixtures of common metal and organic contaminants. For 17 sediments with paired data sets from static-renewal and flow-through tests, the best prediction of toxicity to reproduction was achieved using a 2-parameter MLR that included hazard quotients for sediment contaminants and toxic units for dissolved metals (r2 = 0.892). The inclusion of particle size, organic carbon and acid-volatile sulfide did not improve toxicity predictions, despite these parameters being recognised as modifying contaminant bioavailability. The use of dilute-acid-extractable metal concentrations in place total recoverable metal concentrations did not improve the predictions. The study also confirmed that sediments existing within the estuarine and marine bays of Sydney Harbour pose significant risks of adverse effects to benthic organisms.

Remediation criteria for gasworks-impacted sediments: Assessing the effects of legacy hydrocarbons and more recent metal contamination.

Historical contamination of sediments from industries that commenced before environmental regulations were commonplace is prevalent in many large cities. This contamination is frequently overlain and mixed with more recent urban contamination. The remediation of contaminated sites is often a very expensive exercise and the final remediation criteria often reflect a trade-off between protecting human and ecological health and the finances of those deemed responsible for the site clean-up. In this study, we describe an assessment of estuarine sediments impacted historically by contamination from a gasworks site. The major historical sediment contaminants included polycyclic aromatic hydrocarbons (PAHs) and other petroleum-related hydrocarbons (TRHs). Elevated concentrations of metals exist throughout the city region due to historical pollution and ongoing urban stormwater discharges. Equilibrium partitioning models were used to consider the influence on the bioavailability of PAHs of both natural sedimentary organic carbon and forms of black carbon (pyrogenic carbon - coal tars, charcoal). The strongest predictor of the observed sublethal toxicity to amphipod and copepod reproduction was a combination of total PAHs and metals (primarily Cu, Pb and Zn). Total PAHs was the strongest predicting variable for toxicity to organism survival. While high total PAH concentrations were attributed to the former gas works, high background concentrations of metals existed throughout much of this region of the estuary. Thus, without remediation at the estuary-scale, resuspension of the surrounding sediments by tidal currents and boat movements is predicted to re-contaminate remediated areas with sediments that may continue to cause chronic toxicity due to metals. The assessment indicated that remedial actions that remove or isolate sediments that caused toxicity to benthic organism survival would lead to significant improvements in ecosystem health, but toxicity to organism reproduction may remain at similar levels that exist throughout much of this region of the estuary due to high metal concentrations.

Development and application of a rapid amphipod reproduction test for sediment-quality assessment.

Melita plumulosa is an epibenthic, detritivorous amphipod native to eastern Australia that has been adopted as a test organism for toxicity evaluations of contaminated estuarine sediments. In the present study, a 13-d amphipod reproduction test was developed that encompasses gametogenesis, fertilization, and embryo development before hatching. The primary endpoints for the test are fecundity (measured as the number of embryos per individual surviving female) and a fecundity index (fecundity multiplied by the stage of embryo development). This new test has been employed to scrutinize the sediments from a metal-contaminated coastal lagoon. Lake Macquarie (NSW, Australia) is a large, saltwater lagoon that has received metal pollution over many decades, leading to a concentration gradient of trace metals, including Pb, Zn, Cd, and Cu, in the sediments. Within one of the northern bays (Warners Bay), the concentrations of these metals either border on or exceed sediment quality guideline values prescribed by Australian and New Zealand Guidelines for Fresh and Marine Water Quality. In trials with the 13-d amphipod reproduction test, Warners Bay sediments significantly reduced fecundity in the test species. Subsequent tests with clean sediments spiked singly with Pb, Zn, or Cu indicated that no single metal was responsible for the observed toxicity in the field sediments. However, sediments spiked with various combinations of Pb, Zn, Cd, and Cu indicated that Zn in combination with one or more of the other metals was responsible for the reproductive toxicity observed in Warners Bay sediments. In all these tests, measured metal concentrations in overlying water and pore water were low, thus confirming that the observed effects on reproduction could be attributed to dietary exposure to metals.

The impact of sediment bioturbation by secondary organisms on metal bioavailability, bioaccumulation and toxicity to target organisms in benthic bioassays: Implications for sediment quality assessment.

Bioturbation alters the properties of sediments and modifies contaminant bioavailability to benthic organisms. These naturally occurring disturbances are seldom considered during the assessment of sediment quality. We investigated how the presence (High bioturbation) and absence (Low bioturbation) of a strongly bioturbating amphipod within three different sediments influenced metal bioavailability, survival and bioaccumulation of metals to the bivalve Tellina deltoidalis. The concentrations of dissolved copper decreased and manganese increased with increased bioturbation. For copper a strong correlation was observed between increased bivalve survival (53-100%) and dissolved concentrations in the overlying water. Increased bioturbation intensity resulted in greater tissue concentrations for chromium and zinc in some test sediments. Overall, the results highlight the strong influence that the natural bioturbation activities from one organism may have on the risk contaminants pose to other organisms within the local environment. The characterisation of field-based exposure conditions concerning the biotic or abiotic resuspension of sediments and the rate of attenuation of released contaminants through dilution or readsorption may enable laboratory-based bioassay designs to be adapted to better match those of the assessed environment.

Challenges for using quantitative PCR test batteries as a TIE-type approach to identify metal exposure in benthic invertebrates.

The epibenthic amphipod Melita plumulosa shows unique gene expression profiles when exposed to different contaminants. We hypothesized that specific changes in transcript abundance could be used in a battery of quantitative polymerase chain reaction (qPCR) assays as a toxicity identification evaluation (TIE)-like approach to identify the most relevant stressor in field-contaminated sediments. To test this hypothesis, seven candidate transcriptomic markers were selected, and their specificity following metal exposure was confirmed. The performance of these markers across different levels of added metals was verified. The ability of these transcripts to act as markers was tested by exposing amphipods to metal-contaminated field-collected sediments and measuring changes in transcript abundance via qPCR. For two of the three sediments tested, at least some of the transcriptomic patterns matched our predictions, suggesting that they would be effective in helping to identify metal exposure in field sediments. However, following exposure to the third sediment, transcriptomic patterns were unlike our predictions. These results suggest that the seven transcripts may be insufficient to discern individual contaminants from complex mixtures and that microarray or RNA-Seq global gene expression profiles may be more effective for TIE. Changes in transcriptomics based on laboratory exposures to single compounds should be carefully validated before the results are used to analyze mixtures.

The mismatch between bioaccumulation in field and laboratory environments: Interpreting the differences for metals in benthic bivalves.

Laboratory-based bioaccumulation and toxicity bioassays are frequently used to predict the ecological risk of contaminated sediments in the field. This study investigates the bioassay conditions most relevant to achieving environmentally relevant field exposures. An identical series of metal-contaminated marine sediments were deployed in the field and laboratory over 31 days. Changes in metal concentrations and partitioning in both sediments and waters were used to interpret differences in metal exposure and bioaccumulation to the benthic bivalve Tellina deltoidalis. Loss of resuspended sediments and deposition of suspended particulate matter from the overlying water resulted in the concentrations of Cu, Pb and Zn (major contaminants) becoming lower in the 1-cm surface layer of field-deployed sediments. Lower exchange rates of overlying waters in the laboratory resulted in higher dissolved metal exposures. The prediction of metal bioaccumulation by the bivalves in field and laboratory was improved by considering the metal partitioning within the surface sediments.

Assessing mechanisms of toxicant response in the amphipod Melita plumulosa through transcriptomic profiling.

This study describes the function of transcripts with altered abundance in the epibenthic amphipod, Melita plumulosa, following whole-sediment exposure to a series of common environmental contaminants. M. plumulosa were exposed for 48 h to sediments spiked and equilibrated with the following contaminants at concentrations predicted to cause sublethal effects to reproduction: porewater ammonia 30 mg L(-1); bifenthrin at 100 µg kg(-1); fipronil at 50 µg kg(-1); 0.6% diesel; 0.3% crude oil; 250 mg Cu kg(-1); 400 mg Ni kg(-1); and 400 mg Zn kg(-1). RNA was extracted and hybridized against a custom Agilent microarray developed for this species. Although the microarray represented a partial transcriptome and not all features on the array could be annotated, unique transcriptomic profiles were generated for each of the contaminant exposures. Hierarchical clustering grouped the expression profiles together by contaminant class, with copper and zinc, the petroleum products and nickel, and the pesticides each forming a distinct cluster. Many of the transcriptional changes observed were consistent with patterns previously described in other crustaceans. The changes in the transcriptome demonstrated that contaminant exposure caused changes in digestive function, growth and moulting, and the cytoskeleton following metal exposure, whereas exposure to petroleum products caused changes in carbohydrate metabolism, xenobiotic metabolism and hormone cycling. Functional analysis of these gene expression profiles can provide a better understanding of modes of toxic action and permits the prediction of mixture effects within contaminated ecosystems.

454 pyrosequencing-based analysis of gene expression profiles in the amphipod Melita plumulosa: transcriptome assembly and toxicant induced changes.

Next generation sequencing using Roche's 454 pyrosequencing platform can be used to generate genomic information for non-model organisms, although there are bioinformatic challenges associated with these studies. These challenges are compounded by a lack of a standardized protocol to either assemble data or to evaluate the quality of a de novo transcriptome. This study presents an assembly of the control and toxicant responsive transcriptome of Melita plumulosa, an Australian amphipod commonly used in ecotoxicological studies. RNA was harvested from control amphipods, juvenile amphipods, and from amphipods exposed to either metal or diesel contaminated sediments. This RNA was used as the basis for a 454 based transcriptome sequencing effort. Sequencing generated 1.3 million reads from control, juvenile, metal-exposed and diesel-exposed amphipods. Different read filtering and assembly protocols were evaluated to generate an assembly that (i) had an optimal number of contigs; (ii) had long contigs; (iii) contained a suitable representation of conserved genes; and (iv) had long ortholog alignment lengths relative to the length of each contig. A final assembly, generated using fixed-length trimming based on the sequence quality scores, followed by assembly using the MIRA algorithm, produced the best results. The 26,625 contigs generated via this approach were annotated using Blast2GO, and the differential expression between treatments and control was determined by mapping with BWA followed by DESeq. Although the mapping generated low coverage, many differentially expressed contigs, including some with known developmental or toxicological function, were identified. This study demonstrated that 454 pyrosequencing is an effective means of generating reference transcriptome information for organisms, such as the amphipod M. plumulosa, that have no genomic information available in databases or in closely related sequenced species. It also demonstrated how optimization of read filtering protocols and assembly approaches changes the utility of results obtained from next generation sequencing studies, and establishes criteria to determine the quality of a de novo assembly in species lacking a reference genome. This new transcriptomic knowledge provides the genomic foundation for the creation of microarray and qPCR assays, serving as a reference transcriptome in future RNAseq studies, and allowing both the biology and ecotoxicology of this organism to be better understood. This approach will allow genomics-based methodology to be applied to a wider range of environmentally relevant species.

Incorporating bioavailability into management limits for copper in sediments contaminated by antifouling paint used in aquaculture.

Although now well embedded within many risk-based sediment quality guideline (SQG) frameworks, contaminant bioavailability is still often overlooked in assessment and management of contaminated sediments. To optimise management limits for metal contaminated sediments, we assess the appropriateness of a range methods for modifying SQGs based on bioavailability considerations. The impairment of reproduction of the amphipod, Melita plumulosa, and harpacticoid copepod, Nitocra spinipes, was assessed for sediments contaminated with copper from antifouling paint, located below aquaculture cages. The measurement of dilute acid-extractable copper (AE-Cu) was found to provide the most useful means for monitoring the risks posed by sediment copper and setting management limits. Acid-volatile sulfide was found to be ineffective as a SQG-modifying factor as these organisms live mostly at the more oxidised sediment water interface. SQGs normalised to %-silt/organic carbon were effective, but the benefits gained were too small to justify this approach. The effectiveness of SQGs based on AE-Cu was attributed to a small portion of the total copper being present in potentially bioavailable forms (typically<10% of the total). Much of the non-bioavailable form of copper was likely present as paint flakes in the form of copper (I) oxide, the active ingredient of the antifoulant formulation. While the concentrations of paint-associated copper are very high in some sediments, as the transformation of this form of copper to AE-Cu appears slow, monitoring and management limits should assess the more bioavailable AE-Cu forms, and further efforts be made to limit the release of paint particles into the environment.

Performance and sensitivity of rapid sublethal sediment toxicity tests with the amphipod Melita plumulosa and copepod Nitocra spinipes.

Sublethal whole-sediment toxicity tests are an important tool for assessing the potential effects of contaminated sediments. However, the longer duration required for evaluating potential chronic effects may increase endpoint variability and test costs compared to survival endpoints. In the present study we compare the performance and sensitivity to contaminants of 10-d sublethal sediment toxicity tests with the amphipod Melita plumulosa and harpacticoid copepod Nitocra spinipes. For both tests, sublethal effects were consistently observed when sediment contaminant concentrations exceeded sediment quality guideline (SQG) concentrations. The response of these bioassays in metal-contaminated sediments was shown to conform ideally with respect to the mean SQG quotient calculated on the basis of the Australian and New Zealand lower SQG trigger value, with toxicity being observed only in those sediments where the mean quotient exceeded one. Better predictions of nontoxicity were obtained when dilute acid-extractable rather than total metal concentrations were used. Using the upper SQG, toxicity frequently occurred at mean quotients below one. The effects were generally consistent with predictions from the acid-volatile sulfide and simultaneously extracted metal model. Effects on reproduction of M. plumulosa were detected for sediments that did not cause effects on survival and highlighted the environmental relevance and importance of using these sublethal endpoints. When using four replicates for M. plumulosa and five replicates for N. spinipes, the endpoint variability (standard error) was less than 10%. Variations in sediment particle size and organic carbon content did not affect endpoint variability. Both species are relatively easily cultured in the laboratory, and the estimated effort and cost of achieving the sublethal endpoints is 1.5 times that of the acute survival test endpoints.

Guidelines for copper in sediments with varying properties.

A major weakness of sediment quality guidelines (SQGs) is their poor ability to predict how toxicity thresholds change for different sediment types. Using species sensitivity distributions (SSDs) of copper effects data, new guidelines were derived for copper in non-sulfidic marine sediments in which organic carbon (OC) and particle size strongly influence copper bioavailability. The derived SQGs varied in a predictable manner with changes in sediment particle size and organic carbon (OC), and were shown to offer a significant improvement on the existing 'single value' SQG. Adequate protection for all benthic organisms is expected to be achieved for a OC-normalised copper concentration of 3.5 mg Cu g(-1) OC in the <63 µm sediment fraction. For short-term exposures, the equivalent acute guideline is 11 mg<63 µm Cu g(-1) OC. The new SQGs incorporate a high degree of conservatism owing to the use of copper-spiked sediments and laboratory-based bioassays that were expected to result in greater metal exposure of organisms to bioavailable copper than would be expected for field-contaminated sediments with similar total copper concentrations. SQGs that vary with sediment properties were prepared in an easily referenced tabular format.