Effect of Dissolved Organic Matter Concentration and Source on the Chronic Toxicity of Copper and Nickel Mixtures to Chlorella sp.

There have been limited studies on the effects of toxicity-modifying factors, such as dissolved organic matter (DOM), on the toxicity of metal mixtures to aquatic biota. The present study investigated the effects of DOM concentration (low, 2.8 ± 0.1 mg C/L; high, 11 ± 1.0 mg C/L) and DOM source (predominantly terrestrial or microbial) on the chronic toxicity of copper (Cu) and nickel (Ni) binary mixtures to the green freshwater microalga Chlorella sp. This was assessed by using a full factorial design of 72-h growth inhibition bioassays. Measured algal growth rate was compared with growth predicted by the concentration addition and independent action reference models. Model predictions were based on concentrations of dissolved metals, labile metals (measured by diffusive gradients in thin films [DGT]), and calculated free metal ions (determined by the Windermere Humic Aqueous Model). Copper/Ni mixture toxicity was synergistic to Chlorella sp. in the absence of added DOM, with evidence of metal concentration-dependent toxicity at low effect concentrations. As DOM concentration increased, the mixture interaction changed from synergism to noninteraction or antagonism depending on the metal speciation method used. The DOM source had no significant effect on mixture interaction when based on dissolved and free metal ion concentrations but was significantly different when based on DGT-labile metal concentrations. Ratio-dependent mixture interaction was observed in all treatments, with increased deviation from the reference model predictions as the mixture changed from Ni- to Cu-dominated. The present study demonstrated that both DOM concentration and source can significantly change metal mixture toxicity interactions and that these interactions can be interpreted differently depending on the metal speciation method used. Environ Toxicol Chem 2021;40:1908-1918. © 2021 SETAC.

Metabarcoding Reveals Changes in Benthic Eukaryote and Prokaryote Community Composition along a Tropical Marine Sediment Nickel Gradient.

The Southeast Asia and Melanesia region has extensive nickel (Ni)-rich lateritic regoliths formed from the tropical weathering of ultramafic rocks. As the global demand for Ni continues to rise, these lateritic regoliths are increasingly being exploited for their economic benefit. Mining of these regoliths contributes to the enrichment of coastal sediments in trace metals, especially Ni. The present study used high-throughput sequencing (metabarcoding) to determine changes in eukaryote (18s v7 recombinant DNA [rDNA] and diatom-specific subregion of the 18s v4 rDNA) and prokaryote (16s v4 rDNA) community compositions along a sediment Ni concentration gradient offshore from a large lateritized ultramafic regolith in New Caledonia (Vavouto Bay). Significant changes in the eukaryote, diatom, and prokaryote community compositions were found along the Ni concentration gradient. These changes correlated most with the dilute-acid extractable concentration of Ni in the sediments, which explained 26, 23, and 19% of the variation for eukaryote, diatom, and prokaryote community compositions, respectively. Univariate analyses showed that there was no consistent change in indices of biodiversity, evenness, or richness. Diatom richness and diversity did, however, decrease as sediment acid extractable-Ni concentrations increased. Threshold indicator taxa analysis was conducted separately for each of the 3 targeted genes to detect changes in taxa whose occurrences decreased or increased along the acid extractable-Ni concentration gradient. Based on these data, 46 mg acid extractable-Ni/kg was determined as a threshold value where sensitive species began to disappear. In the case of the estuarine sediments offshore from lateritized ultramafic regolith in New Caledonia, this is recommended as an interim threshold value until further lines of evidence can contribute to a region-specific Ni sediment quality guideline value. Environ Toxicol Chem 2021;40:1894-1907. © 2021 SETAC.

Speciation of nickel and its toxicity to Chlorella sp. in the presence of three distinct dissolved organic matter (DOM).

Nickel is often a metal of interest in regulatory settings given its increasing prevalence in disturbed freshwaters and as a known toxicant to fish and algae. Dissolved organic matter (DOM) is a toxicity modifying factor for nickel and a ubiquitous water physicochemical parameter. This study investigated the effect of DOM concentration and source on the chronic toxicity of nickel to Chlorella sp. using three DOM at two concentrations (3.1 ± 1.8 and 12 ± 1.3 mg C/L). Nickel toxicity to Chlorella sp. was not strongly influenced by DOM concentration. In the absence of DOM, the 72-h EC50 for Chlorella sp. was 120 µg Ni/L. In the low DOM treatment, nickel toxicity was either unchanged or slightly increased (87-140 µg Ni/L) and unchanged or slightly decreased in the high DOM treatment (130-240 µg Ni/L). DOM source also had little effect on nickel toxicity, the largest differences in nickel toxicity occurring in the high DOM treatment. Labile nickel (measured by diffusive gradients in thin-films, DGT) followed strong linear relationships with dissolved nickel (R2 > 0.97). DOM concentration and source had limited effect on DGT-labile nickel. DGT-labile nickel decreased with increasing DOM concentration for only one of the three DOM. Modelled labile nickel concentrations (expressed as maximum dynamic concentrations, cdynmax) largely agreed with DGT-labile nickel and suggested that toxicity is explained by free Ni2+ concentrations. This study confirms that nickel toxicity is largely unaffected by DOM concentration or source and that both measured (DGT) and modelled (cdynmax and free Ni2+) nickel concentrations can explain nickel toxicity.

The Diffusive Gradients in Thin Films Technique Predicts Sediment Nickel Toxicity to the Amphipod Melita plumulosa.

The geographical shift of nickel mining to small island countries of the Southeast Asia and Melanesia region has produced a need to assess the environmental risk associated with increased sediment nickel exposure to benthic estuarine/marine biota. Chemical measurements of nickel concentration and potential bioavailability, including the use of diffusive gradients in thin films (DGT), were compared to effects on 10-d reproduction of the epibenthic estuarine/marine amphipod Melita plumulosa in nickel-spiked sediments and field-contaminated sediments with different characteristics. The 10% effect concentrations (EC10s) for amphipod reproduction ranged from 280 to 690 mg/kg total recoverable nickel, from 110 to 380 mg/kg dilute acid-extractable nickel, and from 34 to 87 µg Ni/m2 /h DGT-labile nickel flux. Nickel bioavailability was lower in sediments with greater total organic carbon, clay content, and percentage of fine particles. Measurements of DGT-labile nickel flux at the sediment-water interface integrated exposure to nickel from porewater, overlying water, and ingested sediment exposure pathways and were found to have the strongest relationship with the biological response. At most, there was a 29% reduction in 10-d M. plumulosa reproduction relative to the control when exposed to nickel from field-contaminated sediments collected from nickel laterite mining regions of New Caledonia. The DGT technique can be used as a complementary tool to measure the bioavailability of nickel in estuarine/marine sediments, especially sediments that are in nickel laterite mining regions where there are no or few toxicity data available for determining biological effects on local species. Based on the combined data set of the 3 nickel-spiked sediments a DGT-labile nickel EC10 threshold of 50 (30-69) µg Ni/m2 /h was determined. Environ Toxicol Chem 2021;40:1266-1278. © 2020 SETAC.

Effects of dissolved nickel and nickel-contaminated suspended sediment on the scleractinian coral, Acropora muricata.

Intensification of lateritic nickel mining in Southeast Asia and Melanesia potentially threatens coastal ecosystems from increased exposure to nickel and suspended sediment. This study investigated the response of Acropora muricata when exposed to either dissolved nickel, clean suspended sediment or nickel-contaminated suspended sediment for 7 days, followed by a 7-d recovery period. Significant bleaching and accumulation of nickel in coral tissue was observed only after exposure to high dissolved nickel concentrations and nickel-spiked suspended sediment. No effect on A. muricata was observed from exposure to a particulate-bound nickel concentration of 60 mg/kg acid-extractable nickel at a suspended sediment concentration of 30 mg/L TSS. This study demonstrates that bioavailability of nickel associated with suspended sediment exposure plays a key role in influencing nickel toxicity to corals. These findings assist in assessments of risk posed by increasing nickel mining activities on tropical marine ecosystems.

The effect of dissolved nickel and copper on the adult coral Acropora muricata and its microbiome.

The potential impacts of mining activities on tropical coastal ecosystems are poorly understood. In particular, limited information is available on the effects of metals on scleractinian corals which are foundation species that form vital structural habitats supporting other biota. This study investigated the effects of dissolved nickel and copper on the coral Acropora muricata and its associated microbiota. Corals collected from the Great Barrier Reef were exposed to dissolved nickel (45, 90, 470, 900 and 9050 µg Ni/L) or copper (4, 11, 32 and 65 µg Cu/L) in flow through chambers at the National Sea Simulator, Townsville, Qld, Australia. After a 96-h exposure DNA metabarcoding (16S rDNA and 18S rDNA) was undertaken on all samples to detect changes in the structure of the coral microbiome. The controls remained healthy throughout the study period. After 36 h, bleaching was only observed in corals exposed to 32 and 65 µg Cu/L and very high nickel concentrations (9050 µg Ni/L). At 96 h, significant discolouration of corals was only observed in 470 and 900 µg Ni/L treatments, the highest concentrations tested. While high concentrations of nickel caused bleaching, no changes in the composition of their microbiome communities were observed. In contrast, exposure to copper not only resulted in bleaching, but altered the composition of both the eukaryote and bacterial communities of the coral's microbiomes. Our findings showed that these effects were only evident at relatively high concentrations of nickel and copper, reflecting concentrations observed only in extremely polluted environments. Elevated metal concentrations have the capacity to alter the microbiomes which are inherently linked to coral health.

Assessing the chronic toxicity of nickel to a tropical marine gastropod and two crustaceans.

The mining and processing of nickel ores from tropical regions contributes 40% of the global supply. The potential impact of these activities on tropical marine ecosystems is poorly understood. Due to the lack of ecotoxicity data for tropical marine species, there is currently no available water quality guideline value for nickel that is specific to tropical species. In this study, we investigated the toxicity of nickel to three tropical marine invertebrates, the gastropod Nassarius dorsatus, the barnacle Amphibalanus amphitrite, and the copepod Acartia sinjiensis. All toxicity tests used chronic endpoints, namely larval growth, metamorphosis (transition from nauplii to cyprid larvae) and larval development for the snail, barnacle and copepod respectively. Toxicity tests were carried out under environmentally relevant conditions (i.e. 27-30ᵒC, salinity 34-36‰, pH 8.1-8.4). Copper was also tested for quality assurance purposes and to allow for comparisons with previous studies. The copepod was the most sensitive species to nickel, with development inhibited by 10% (EC10) at 5.5 (5.0-6.0) µg Ni/L (95% confidence limits (CL)). Based on EC10 values, the gastropod and barnacle showed similar sensitivities to nickel with growth and metamorphosis inhibited by 10% at 64 (37-91) µg Ni/L and 67 (53-80) µg Ni/L, respectively. Based on existing data available in the literature, the copepod A. sinjiensis is so far the most sensitive tropical marine species to nickel. This study has provided high quality data which will contribute to the development of a water quality guideline value for nickel in tropical marine waters. A species sensitivity distribution of chronic nickel toxicity used the data generated in this paper supplemented by available literature data, comprising 12 species representing 6 taxonomic groups. A 5% hazard concentration (HC5) was determined as 8.2 µg/L Ni.

Inhibition in fertilisation of coral gametes following exposure to nickel and copper.

The mining and production of nickel in tropical regions have the potential to impact on ecologically valuable tropical marine ecosystems. Currently, few data exist to assess the risks of nickel exposure to tropical ecosystems and to derive ecologically relevant water quality guidelines. In particular, data are lacking for keystone species such as scleractinian corals, which create the complex structural reef habitats that support many other marine species. As part of a larger study developing risk assessment tools for nickel in the tropical Asia-Pacific region, we investigated the toxicity of nickel on fertilisation success in three species of scleractinian corals: Acropora aspera, Acropora digitifera and Platygyra daedalea. In the literature, more data are available on the effects of copper on coral fertilisation, so to allow for comparisons with past studies, the toxicity of copper to A. aspera and P. daedalea was also determined. Overall, copper was more toxic than nickel to the fertilisation success of the species tested. Acropora aspera was the most sensitive species to nickel (NOEC < 280µg Ni/L), followed by A. digitifera with an EC10 of 2000µg Ni/L and P. daedalea (EC10 > 4610µg Ni/L). Acropora aspera was also the more sensitive species to copper with an EC10 of 5.8µg Cu/L. The EC10 for P. daedalea was 16µg Cu/L, similar to previous studies. This is the first time that the toxicity of nickel on fertilisation success in Acropora species has been reported, and thus provides valuable data that can contribute to the development of reliable water quality guidelines for nickel in tropical marine waters.

A review of nickel toxicity to marine and estuarine tropical biota with particular reference to the South East Asian and Melanesian region.

The South East Asian Melanesian (SEAM) region contains the world's largest deposits of nickel lateritic ores. Environmental impacts may occur if mining operations are not adequately managed. Effects data for tropical ecosystems are required to assess risks of contaminant exposure and to derive water quality guidelines (WQG) to manage these risks. Currently, risk assessment tools and WQGs for the tropics are limited due to the sparse research on how contaminants impact tropical biota. As part of a larger project to develop appropriate risk assessment tools to ensure sustainable nickel production in SEAM, nickel effects data were required. The aim of this review was to compile data on the effects of nickel on tropical marine, estuarine, pelagic and benthic species, with a particular focus on SEAM. There were limited high quality chronic nickel toxicity data for tropical marine species, and even fewer for those relevant to SEAM. Of the data available, the most sensitive SEAM species to nickel were a sea urchin, copepod and anemone. There is a significant lack of high quality chronic data for several ecologically important taxonomic groups including cnidarians, molluscs, crustaceans, echinoderms, macroalgae and fish. No high quality chronic nickel toxicity data were available for estuarine waters or marine and estuarine sediments. The very sparse toxicity data for tropical species limits our ability to conduct robust ecological risk assessment and may require additional data generation or read-across from similar species in other databases (e.g. temperate) to fill data gaps. Recommendations on testing priorities to fill these data gaps are presented.

Copper Uptake, Intracellular Localization, and Speciation in Marine Microalgae Measured by Synchrotron Radiation X-ray Fluorescence and Absorption Microspectroscopy.

Metal toxicity to aquatic organisms depends on the speciation of the metal and its binding to the critical receptor site(s) (biotic ligand) of the organism. The intracellular nature of the biotic ligand for Cu in microalgal cells was investigated using the high elemental sensitivity of microprobe synchrotron radiation X-ray fluorescence (SR-XRF) and X-ray absorption near-edge spectroscopy (XANES). The marine microalgae, Ceratoneis closterium, Phaeodactylum tricornutum, and Tetraselmis sp. were selected based on their varying sensitivities to Cu (72-h 50% population growth inhibitions of 8-47 µg Cu/L). Intracellular Cu in control cells was similar for all three species (2.5-3.2 × 10(-15) g Cu/cell) and increased 4-fold in C. closterium and Tetraselmis sp. when exposed to copper, but was unchanged in P. tricornutum (72-h exposure to 19, 40, and 40 µg Cu/L, respectively). Whole cell microprobe SR-XRF identified endogenous Cu in the central compartment (cytoplasm) of control (unexposed) cells. After Cu exposure, Cu was colocated with organelles/granules dense in P, S, Ca, and Si and this was clearly evident in thin sections of Tetraselmis sp. XANES indicated coexistence of Cu(I) and Cu(II) in control and Cu-exposed cells, with the Cu ligand (e.g., phytochelatin) in P. tricornutum different from that in C. closterium and Tetraselmis sp. This study supports the hypothesis that Cu(II) is reduced to Cu(I) and that polyphosphate bodies and phytochelatins play a significant role in the internalization and detoxification of Cu in marine microalgae.

Copper-induced changes in intracellular thiols in two marine diatoms: Phaeodactylum tricornutum and Ceratoneis closterium.

Phytochelatins and glutathione (reduced (GSH) and oxidised (GSSG)) are important intracellular ligands involved in metal sequestration and detoxification in algae. Intracellular ratios of GSH:GSSG are sensitive indicators of metal stress in algae, and like phytochelatin production are influenced by metal speciation, concentration, exposure time and the biological species. This study investigated the effect of copper exposure on phytochelatin and glutathione content in two marine diatoms Phaeodactylum tricornutum and Ceratoneis closterium at various time intervals between 0.5 and 72h. Liberation of cellular glutathione and phytochelatins was optimised using freeze/thaw cycles and chemical extraction, respectively. Extracted phytochelatins were derivatised (by fluorescent tagging of thiol compounds), separated and quantified using HPLC with fluorescence detection. Glutathione ratios were determined using a commercially available kit, which uses the enzyme glutathione reductase to measure total and oxidised glutathione. Despite similarities in size and shape between the two diatoms, differences in internalised copper, phytochelatin production (both chain length and quantity) and reduced glutathione concentrations were observed. P. tricornutum maintained reduced glutathione at between 58 and 80% of total glutathione levels at all time points, which would indicate low cellular stress. In C. closterium reduced glutathione constituted <10% of total glutathione after 48h. P. tricornutum also produced more phytochelatins and phytochelatins of longer chain length than C. closterium despite the latter species internalising significantly more copper.

Optimization of colorimetric DET technique for the in situ, two-dimensional measurement of iron(II) distributions in sediment porewaters.

The recently developed colorimetric diffusive equilibration in thin films (DET) technique for the in situ, high-resolution measurement of iron(II) in marine sediments is optimized to allow measurement of the higher iron concentrations typical of freshwater sediment porewaters. Computer imaging densitometry (CID) is used to analyze the retrieved samplers following exposure to ferrozine, a colorimetric reagent selective for iron(II). The effect of ferrozine concentration, image processing parameters and ionic strength are investigated to improve the applicability of this technique to a wider range of aquatic systems than reported in the first publications of this approach. The technique was optimized to allow detection of up to 2,000 µmol L(-1) iron(II), a four-fold increase on the previous upper detection limit of 500 µ mol L(-1). The CID processing of the scanned color image was also optimized to adjust the sensitivity of the assay as required; by processing the image with different color channel filters, the sensitivity of the assay can be optimized for lower concentrations (up to 100 µmol L(-1)) or higher concentrations (up to 2,000 µmol L(-1)) of iron(II), depending on the specific site characteristics. This process does not require separate sampling probes or even separate scans of the DET gels as the color filter and grayscale conversion is done post-image capture. The optimized technique is very simple to use and provides highly representative, high-resolution (1mm) two-dimensional distributions of iron(II) in sediment porewaters. The detection limit of the optimized technique was 4.1±0.3 µmol L(-1) iron(II) and relative standard deviations were less than 6%.

The Gladstone (Australia) oil spill - impacts on intertidal areas: baseline and six months post-spill.

In January 2006, 25 tonnes of heavy fuel oil spilled into the Port of Gladstone in Queensland, Australia, from the breached hull of a bulk carrier ship. While approximately 18 tonnes of the oil was recovered, a certain amount of oil was deposited in the intertidal areas of Port Curtis leaving a highly visible, viscous residue. The objectives of this research were to assess the short-term (one month post-spill) and medium-term (six months post-spill) impacts on the intertidal habitat. Sediment polycyclic aromatic hydrocarbons (PAHs) and metal concentrations, mangrove communities and intertidal macroinvertebrates were assessed at oil impacted sites, adjacent sites which were not visibly impacted and reference sites which were located outside the recorded distribution of the oil spill. At one month post-spill, highest PAH concentrations were found at the impacted sites, with concentrations of some PAHs exceeding Australian and New Zealand sediment quality guidelines (SQG) [ANZECC/ARMCANZ, 2000. Sediment Quality Guidelines. Australia and New Zealand Environment and Conservation Council and Agriculture and Resource Management Council of Australia and New Zealand]. However, by six months post-spill PAH concentrations had significantly decreased. PAH concentrations tended to be higher in the back (upper) intertidal zone than at the front of the mangrove stand, and sediment cores indicated that PAH contaminants had remained in the top 4cm of the sediment. These results indicate that the overall decreased PAH concentrations are likely to be due to evaporation, photoxidation and tidal flushing of the residual oil in these impacted sites. During the initial survey, the impact site contained very few or no crabholes in the high intertidal area, indicating a low crab density in comparison to reference sites. However, at six months post-spill mangrove crab communities appeared to be fully recovered with crabhole densities in impact sites similar to reference sites. While little immediate impact was evident on mangrove parameters, significantly higher seedling mortality and defoliation was apparent at six months post-spill, particularly in the upper intertidal zone. Intertidal macrobenthic communities did not appear to be impacted, either immediately or at six months post-spill. Monitoring of these oil impacted intertidal areas will continue to examine the long-term impacts/recovery from the oil spill.