William Maher

Biotransference and biomagnification of selenium copper, cadmium, zinc, arsenic and lead in a temperate seagrass ecosystem from Lake Macquarie Estuary, NSW, Australia

In this study the biotransference of selenium copper, cadmium, zinc, arsenic and lead was measured in a contaminated seagrass ecosystem in Lake Macquarie, NSW, Australia, to determine if biomagnification of these trace metals is occurring and if they reach concentrations that pose a threat to the resident organisms or human consumers. Selenium was found to biomagnify, exceeding maximum permitted concentrations for human consumption within carnivorous fish tissue, the highest trophic level examined. Selenium concentrations measured within carnivorous fish were also above those shown to elicit sub-lethal effects in freshwater fish. As comparisons are made to selenium concentrations known to effect freshwater fish, inferences must be made with caution. There was no evidence of copper, cadmium, zinc or lead biomagnification within the food web examined. Copper, cadmium, zinc and lead concentrations were below concentrations shown to elicit adverse responses in biota. Copper concentrations within crustaceans M. bennettae and P. palagicus were found to exceed maximum permitted concentrations for human consumption. It is likely that copper concentrations within these species were accumulated due to the essential nature of this trace metal for many species of molluscs and crustaceans. Arsenic showed some evidence of biomagnification. Total arsenic concentrations are similar to those found in other uncontaminated marine ecosystems, thus arsenic concentrations are unlikely to cause adverse effects to aquatic organisms. Inorganic arsenic concentrations are below maximum permitted concentrations for human consumption.

Invertebrate biomarkers: links to toxicosis that predict population decline.

The application of biochemical measurements that can be used as individual biomarkers of impaired biological function in invertebrates is reviewed to evaluate whether biochemical biomarkers of aquatic invertebrates can predict changes in natural populations. Biomarkers that measure toxic effects at the molecular level (e.g., the inhibition of brain acetylcholinesterase activity by organophosphorus pesticides) have been shown to provide rapid quantitative predictions of a toxic effect upon individuals in laboratory studies. Such biomarkers should not be used as a replacement for conventional aquatic monitoring techniques, but should be applied as supplementary approaches for demonstrating links between sublethal biochemical and adverse effects in natural populations in field studies. The research challenge for using biomarker measurements in aquatic invertebrates is to predict effects at the population level from effects at the individual level measured upon individuals collected in the field.

Ecological assessment of selenium in the aquatic environment.

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 Background and Need for Workshop . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 Workshop Purpose and Goals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Participation and Format . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Workgroup Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Workgroup 1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Problem formulation: Context for selenium risk assessment . . . . . . . . . . . . . . . . . . . . . 9 Selenium is a global problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Case studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Conceptual model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 How to investigate a potential selenium problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Recommendations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Workgroup 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Environmental partitioning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Workgroup 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 Bioaccumulation and trophic transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 Workgroup 4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 Selenium toxicity to aquatic organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 Workgroup 5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 Risk characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 Importance of problem formulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 Risk characterization: Unique challenges concerning selenium . . . . . . . . . . . . . . . . . . 26 Risk management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Uncertainties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Overall Workshop Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Appendix: SETAC Pellston Workshop Participants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 List of Figures Figure 1 Conceptual model depicting Se dynamics and transfer in aquatic ecosystems . . . . . . . . . . . . .11 Figure 2 Hierarchy of effects across levels of biological organization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12 Figure 3 Potential sources of Se to aquatic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 Figure 4 Selenium species associated with major processes in aquatic systems . . . . . . . . . . . . . . . . . . . . . . . . . . .16 Figure 5 Partitioning of Se among environmental compartments in a typical aquatic system. . . .16 Figure 6 Selenium enrichment and trophic transfer in aquatic food webs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19 Figure 7 Selenium accumulation in different species of algae, invertebrates, and fish . . . . . . . . . . . . . . . .20 Figure 8 Conceptual pathway of Se transfer in aquatic ecosystems and relative certainty with which Se concentrations in environmental compartments can be assessed in making accurate characterizations of risk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28 List of Tables Table 1 Assessment endpoints and measures of exposure and effect for aquatic and aquaticlinked organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14 Table 2 Uncertainties and recommendations for future research pertaining to toxicity of Se species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .24 Ecological Assessment of Selenium in the Aquatic Environment 4

Toxicity, biotransformation, and mode of action of arsenic in two freshwater microalgae (Chlorella sp. and Monoraphidium arcuatum)

The toxicity of As(V) and As(III) to two axenic tropical freshwater microalgae, Chlorella sp. and Monoraphidium arcuatum, was determined using 72-h growth rate-inhibition bioassays. Both organisms were tolerant to As(III) (72-h concentration to cause 50% inhibition of growth rate [IC50], of 25 and 15 mg As[III]/L, respectively). Chlorella sp. also was tolerant to As(V) with no effect on growth rate over 72 h at concentrations up to 0.8 mg/L (72-h IC50 of 25 mg As[V]/L). Monoraphidium arcuatum was more sensitive to As(V) (72-h IC50 of 0.25 mg As[V]/L). An increase in phosphate in the growth medium (0.15-1.5 mg PO4(3-)/L) decreased toxicity, i.e., the 72-h IC50 value for M. arcuatum increased from 0.25 mg As(V)/L to 4.5 mg As(V)/L, while extracellular As and intracellular As decreased, indicating competition between arsenate and phosphate for cellular uptake. Both microalgae reduced As(V) to As(III) in the cell, with further biological transformation to methylated species (monomethyl arsonic acid and dimethyl arsinic acid) and phosphate arsenoriboside. Less than 0.01% of added As(V) was incorporated into algal cells, suggesting that bioaccumulation and subsequent methylation was not the primary mode of detoxification. When exposed to As(V), both species reduced As(V) to As(III); however, only M. arcuatum excreted As(III) into solution. Intracellular arsenic reduction may be coupled to thiol oxidation in both species. Arsenic toxicity most likely was due to arsenite accumulation in the cell, when the ability to excrete and/or methylate arsenite was overwhelmed at high arsenic concentrations. Arsenite may bind to intracellular thiols, such as glutathione, potentially disrupting the ratio of reduced to oxidized glutathione and, consequently, inhibiting cell division.

Selenium in sediments, pore waters and benthic infauna of Lake Macquarie, New South Wales, Australia

Measurements of selenium in sediments and benthic infauna of Lake Macquarie, an estuary on the east coast of Australia, indicate that sediments are a significant source of selenium in the lakes food web. Analysis of surficial sediment samples indicated higher selenium concentrations near what are believed to be the main industrial sources of selenium to the lake: a smelter and a power station. Sediment cores taken from sediments in Mannering Bay, near a power station at Vales Point, contained an average of 12 times more selenium in surficial sections than sediment cores from Nords Wharf, a part of the lake remote from direct inputs of selenium. The highest selenium concentration found in Mannering Bay sediments (17.2 μg/g) was 69 times the apparent background concentration at Nords Wharf (0.25 μg/g). Pore water concentrations in Mannering Bay were also high, up to 5 μg/l compared to those at Nords Wharf which were below detection limits (0.2 μg/l). Selenium concentrations in muscle tissues of three benthic-feeding fish species (Mugil cephalus, Platycephalus fuscus, Acanthopagrus australis) were significantly correlated (p<0.05) with surficial sediment selenium concentration. Selenium concentrations in polychaetes and molluscs of Mannering Bay were up to 58 times higher than those from Nords Wharf. Two benthic organisms, the eunicid polychaete Marphysa sanguinea and the bivalve mollusc Spisula trigonella, were maintained at different densities in selenium-spiked sediments. Both animals accumulated selenium from the spiked sediment, confirming that bioaccumulation from contaminated sediments occurs. Collectively, these data suggest that benthic food webs are important sources of selenium to the fish of Lake Macquarie.

Bioaccumulation of antimony and arsenic in a highly contaminated stream adjacent to the Hillgrove Mine, NSW, Australia

Environmental context. Concern over the presence of antimony (Sb) in the environment because of chemical similarities with arsenic (As) has prompted a need to better understand its environmental behaviour and risks. The present study investigates the bioaccumulation and uptake of antimony in a highly contaminated stream near the Hillgrove antimony–gold mine in NSW, Australia, and reports high Sb (and As) concentrations in many components of the ecosystem consisting of three trophic levels, but limited uptake into aboveground parts of riparian vegetation. The data suggest that Sb can transfer into upper trophic levels of a creek ecosystem, but that direct exposure of creek fauna to creek sediment and soil, water and aquatic autotrophs are more important metalloid uptake routes than exposure via riparian vegetation. Abstract. Bioaccumulation and uptake of antimony (Sb) were investigated in a highly contaminated stream, Bakers Creek, running adjacent to mining and processing of Sb–As ores at Hillgrove Mine, NSW, Australia. Comparisons with arsenic (As) were included owing to its co-occurrence at high concentrations. Mean metalloid creek rhizome sediment concentrations were 777 ± 115 μg g–1 Sb and 60 ± 6 μg g–1 As, with water concentrations at 381 ± 23 μg L–1 Sb and 46 ± 2 μg L–1 As. Antimony and As were significantly elevated in aquatic autotrophs (96–212 μg g–1 Sb and 32–245 μg g–1 As) but Sb had a lower uptake efficiency. Both metalloids were elevated in all macroinvertebrates sampled (94–316 μg g–1 Sb and 1.8–62 μg g–1 As) except Sb in gastropods. Metalloids were detected in upper trophic levels although biomagnification was not evident. Metalloid transfer to riparian vegetation leaves from roots and rhizome soil was low but rhizome soil to leaf As concentration ratios were up to 2–3 times greater than Sb concentration ratios. Direct exposure to the rhizosphere sediments and soils, water ingestion and consumption of aquatic autotrophs appear to be the major routes of Sb and As uptake for the fauna of Bakers Creek.

Arsenic Species Determination in Biological Tissues by HPLC–ICP–MS and HPLC–HG–ICP–MS

The use of high-pressure liquid chromatography coupled directly or by a hydride generation system to an inductively coupled plasma mass spectrometer for the unambiguous measurement of 13 arsenic species in marine biological extracts is described. The use of two chromatography systems; a Supelcosil LC-SCX cation-exchange column eluted with a 20 mM pyridine mobile phase adjusted to pH 2.2 and 2.6 with formic acid, with a flow rate of 1.5 mL min−1 at 40°C, and a Hamilton PRP-X100 anion-exchange column eluted with 20 mM NH4H2PO4 buffer at pH 5.6, with a flow rate of 1.5 mL min−1 at 40°C, was required to separate and quantify cation and anion arsenic species. Under these conditions, arsenous acid could not be separated from other arsenic species and required the use of an additional hydride generation step. Arsenic species concentrations in a locally available Tasmanian kelp (Durvillea potatorum), a certified reference material (DORM-2), and a range of commercially available macroalgae supplements and sushi seaweeds have been measured and are provided for use as in-house quality control samples to assess the effectiveness of sample preparation, extraction, and measurement techniques.

Selenium contamination, redistribution and remobilisation in sediments of Lake Macquarie, NSW

This paper examines the history of selenium pollution in Lake Macquarie, NSW, Australia, and three factors that may aAect the redistribution and remobilisation of particle bound selenium: changes in redox state, bioturbation, and bioaccumulation by macrobenthos and bacteria. Sediment cores were taken from Nords Wharf, a relatively unpolluted area, and from Mannering Bay near the Vales Point coal-fired power station. The age profile at the unpolluted site seems to indicate that mild selenium pollution has been occurring for over 100 years, however, some mixing of the sediments has occurred. At the polluted site, the age profile indicated that major contamination has occurred in the last 30 years, due to a fly ash dam associated with nearby electric power generation facilities. The contamination chronology suggests that remobilisation and reduction processes have aAected the selenium profile. Changing the redox state of Lake Macquarie sediment results in a release of selenium under oxidising conditions and immobilisation under reducing conditions. The sediment-bound selenium was associated with the operationally defined ‘organic/sulfide’ fraction under reducing conditions, and as the redox potential increases this moves into the ‘exchangeable’ and ‘iron/manganese oxyhydroxide’ phases to a limited extent. Bioturbation by the animals Marphysa sanguinea and Spisula trigonella caused increases in the redox potential and pore water selenium concentrations in surficial sediments relative to unbioturbated controls. Both animals accumulated significantly more selenium when exposed to contaminated sediment than when exposed to uncontaminated control sediments. Selenium concentrations in molluscs from Mannering Bay were all significantly higher than those collected from Nords Wharf. Most of the selenium in the mollusc tissues was found to be associated with the protein fraction. Selenium isolated from hydrolysed muscle tissue was not present as selenate or selenite but as selenomethionine and an unidentified compound. Seven types of bacteria were isolated from Lake Macquarie sediment. All seven isolates were able to transform selenite quantitatively to elemental selenium as evidenced by a red precipitate and identified by X-ray diAraction. Six isolates grew on media containing selenate but no elemental selenium was formed. Mass balances showed that for three isolates total selenium was conserved, selenate decreased and selenium (0; II-) increased indicating the production of non-volatile organic selenium compounds. For two isolates both total selenium and selenate decreased with no increase in selenium (0; II-), therefore, loss of selenium occurred from the media.

Low volume microwave digestion for the determination of selenium in marine biological tissues by graphite furnace atomic absorption spectroscopy

A microwave digestion method for the determination of marine biological tissues has been developed to allow determination of selenium in small sample sizes (< 0.1 g). The benefits of this technique include maintaining concentrates in extracts without the subsequent over dilution encountered when using larger vessels, increased sample throughput and reduced loss of volatile material. Freeze dried biological material (< 0.1 g) and nitric acid (1 ml) were placed into 7 ml screw top Teflon vessels which are completely sealed on capping. Two 7 ml vials were placed into larger 120 ml vessels fitted with a Teflon spacer and 10 ml of distilled water. The effects of microwave power and time, and sample mass on selenium recovery from three marine standard reference materials (NIST SRM 1566a Oyster Tissue, NRCC DORM-1 Dogfish Muscle and NRCC TORT-1 Lobster Hepatopancreas) were examined. The optimum conditions: 600 W, 2 min; 0 W, 2 min; 450 W, 45 min, allowed quantitative recoveries of selenium from these and three other standard reference materials (NRCC DOLT-1 Dogfish liver, NIST RM-50 Albacore tuna and IAEA MA-A-2 fish flesh). Studies on sample mass showed that the analysis of sample masses from 0.025 to 0.1 g gave selenium concentrations within the certified range. Six species of selenium: selenite, selenate, selenomethionine, selenocysteine, selenocystamine, and trimethyl selenonium were added to oyster, dogfish, and lobster tissues. Recoveries were near quantitative for all species (94–105%) except trimethyl selenonium (90–101%).

The occurrence, distribution and sources of polycycic aromatic hydrocarbons in the sediments of the Georges River estuary, Australia

Concentrations of 13 polycylic aromatic hydrocarbons (PAH) in sediment samples from the Georges River estuary N.S.W. have been determined by gas chromatography/mass spectrometry (GC-MS). The concentrations of individual PAH were in the range of <0.1-5600 #g kg -m dry weight. The average values of PAH (I I-1300 #g kg-l dry weight) are well above those previously found in other industralized urban areas of Australia. The highest total PAH concentrations in sediments are found in a section of the river which also has the highest concentrations of silt, clay and organic matter, indicating that the PAH distribution may be controlled by sedimentation processes. Examination of the alkylated and non-alkylated PAH distributions in sediment extracts suggested that the main source of PAH is from combustion products, possibly associated with air particles, with point source contributions of two ring PAH from marinas and other boating activities at some sites.