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Ecotoxicological tests were done at Davis and Casey Stations in 2009/10, 2010/11 and 2011/12 summer seasons under AAS Project 3054 to test the sensitivity of near-shore marine invertebrates to fuels in seawater. The three fuel types used in this project were: Special Antarctic Blend diesel (SAB), Marine Gas Oil diesel (MGO) and an intermediate grade (180) of marine bunker fuel oil (IFO). This dataset contains the results of tests with the near-shore amphipod species Paramoera walkeri exposed to WAFs of SAB, MGO and IFO 180 (specified below) conducted at Davis Station in 2009/10 summer (Season 1). Test treatments were obtained by experimentally mixing fuel and seawater in temperature control cabinets at -1°C to prepare a mixture of fuel hydrocarbons in filtered seawater (FSW) termed the water accommodated fraction (WAF). WAF was produced by adding fuel to seawater in 5 L or 10 L Pyrex glass bottles using a ratio of 1:40 fuel : FSW. This mixture was stirred at slow speed with minimal vortex on a magnetic stirrer. The water portion was then drawn from beneath the fuel. Test treatments consisted of undiluted 100% WAF and dilutions of 10% and 1% of WAFs in FSW. Toxicity tests were conducted in open glass vessels in temperature controlled cabinets. Mortality and/or sub-lethal effects were observed at endpoints of 24 h, 48 h, 96 h, 7 d, 14 d, and 21 d. Treatments were renewed at 7 d intervals. Water quality data was collected at each water change. Hydrocarbon concentrations in WAFs were determined from replicate experiments to measure THC in WAFs over time (Dataset AAS_3054_THC_WAF). WAF exposure concentrations for each test endpoint were derived from these hydrocarbon tests to account for depletion of hydrocarbons from test treatments and any renewal of treatments. An integrated concentration was calculated from measured hydrocarbon concentrations weighted to time. These integrated THC concentrations for endpoints from 24h to 21d are contained in dataset AAS_3054_THC_WAF_integrated_conc_09_10 and are the exposure concentrations used for analysis of sensitivity. Species tested; Paramoera walkeri amphipod; adults This dataset consists of Excel spreadsheets. The file name code for invertebrate tests is; Project number_Season_Taxa_Test name Eg AAS_3054_09_10_amphipod_1PWA1 Project number : AAS_3054 Season : 2009/10 season Taxa: amphipod Test name: 1 for Season 1, PW for genus and species, A for adult, 1 for Test 1 Spreadsheets contain the results of tests with this species. Where replicate tests were conducted, each experiment is on a separate spreadsheet. The worksheet labelled 'Test conditions' shows details of Test name, dates, animal collection details, laboratory holding conditions, details of water accommodated fractions (WAF), test conditions, scoring criteria and water quality data. The worksheet labelled 'Counts' has columns for Replicate number and columns with the Score for all the animals in that replicate at every time endpoint. A full description of the scoring criteria is on the 'Test conditions' worksheet. Totals, means and standard deviations are calculated for each treatment. The worksheet labelled 'Totals, means, percent, StDev' has calculations of Survival, Unaffected, including mean and standard deviation, Percent Survival and Unaffected including means and standard deviation. Amphipod tests also show the Total number of moults in each treatment. Samples were collected at the following locations: - Airport Beach, Davis, Vestfold Hills
This metadata record contains the results of bioassays conducted to characterise the response of Antarctic nearshore marine invertebrates to hydrocarbon contaminants in fuels commonly used in Antarctica. AAS Project 3054. The results of Season 2 and Season 3 amphipod tests are in this dataset. Ecotoxicological bioassays were conducted at Davis and Casey Stations in 2009/10, 2010/11 and 2011/12 summer seasons to test the sensitivity of marine invertebrates to fuels in seawater. The three fuel types used in this project were: Special Antarctic Blend diesel (SAB), Marine Gas Oil diesel (MGO) and an intermediate grade (180) of marine bunker Fuel Oil (IFO). Test treatments were obtained by experimentally mixing fuel and seawater in temperature control cabinets at -1 degrees C to prepare a mixture of fuel hydrocarbons in filtered seawater (FSW) termed the Water Accommodated Fraction (WAF). WAF was produced by adding fuel to seawater in 5 L or 10 L Pyrex glass bottles using a ratio of 1:25 Fuel : FSW. This mixture was stirred at slow speed with minimal vortex for 18 h on a magnetic stirrer. The mixture was settled for 6 h before the water portion was drawn from beneath the fuel. This dataset contains the results of ecotoxicological bioassays with near-shore marine amphipod species exposed to WAFs of SAB WAF, MGO WAF and IFO WAF (specified above). Experimental treatments consisted of undiluted 100% WAF and dilutions of 10% and 1% of WAFs in FSW, to test the toxicity of water accommodated fractions of these three fuels on Antarctic marine invertebrates. The majority of experiments tested WAFs of each of the three fuels, although one tested SAB only due to limited supply of test organisms. Bioassays were conducted in open vessels (glass jars or beakers) in temperature controlled cabinets. Mortality and/or sub-lethal effects were observed at endpoints of 24 h, 48 h, 96 h, 7 d, 8 d, 10 d, 12 d, 14 d, 16 d and 21 d. New WAF solutions were prepared at 4 d intervals to replenish the experimental treatments. Deionised water was added to test solutions as required to maintain test solution volume and salinity. Water quality data was collected at each water change. Hydrocarbon concentrations in WAFs were determined from replicate experiments to measure THC in WAFs over time (Dataset AAS_3054_THC_WAF). WAF exposure concentrations for each bioassay endpoint were derived from these hydrocarbon tests. An integrated concentration was calculated from measured hydrocarbon concentrations weighted to time. Calculations account for depletion of hydrocarbons from test treatments and any renewal of treatments. These integrated THC concentrations for endpoints from 24h to 21d are contained in dataset AAS_3054_THC_WAF_integ_conc_10_11_12. This dataset consists of Excel spreadsheets. The file name code for invertebrate bioassays is; Project number_Season_Taxa_Test name Eg AAS_3054_10_11_amphipod_2PWA1 Project number : AAS_3054 Season : 2010/11 season Taxa: amphipod Test name:2 for Season 2, PW for genus and species, A for adult, 1 for Test 1 Bioassay spreadsheets contain the results of bioassays for a species. Where replicate tests were conducted, each experiment is on a separate spreadsheet. The worksheet labelled "Test conditions" shows details of Test name, dates, animal collection details, laboratory holding conditions, details of water accommodated fractions (WAF), bioassay conditions, scoring criteria and water quality data. The worksheet labelled "Counts" has columns for Replicate number and columns with the Score for all the animals in that replicate at every time endpoint. A full description of the scoring criteria is on the "Test conditions" worksheet. Totals, means and standard deviations are calculated for each treatment. The worksheet labelled "Totals, means, percent, StDev" has calculations of Survival, Unaffected, including mean and standard deviation, Percent Survival and Unaffected including means and standard deviation. Also included is column for the Total number of moults in each treatment. During the research to obtain early life stages of invertebrates for experiments, the number of Paramoera walkeri amphipod neonates per female, the timing of their release from the brood pouch and their early growth rate were recorded. These data are also included in AAS_3054_10_11_PW_neonates Samples were collected from: Ellis Narrows, Vestfold Hills Airport Beach, Davis, Vestfold Hills Prydz Bay, Davis (Between Anchorage Island and Bluff Island) Bailey Peninsula, Windmill Islands
This metadata record contains the results from bioassays conducted to show the response of the Antarctic gastropod, Skenella palludinoides to contamination from combinations of Special Antarctic Blend (SAB) diesel, chemically dispersed with fuel dispersant Ardrox 6120. Fuel only water accommodated fractions (WAF), chemically enhanced water accommodated fractions (CEWAF) and dispersant only treatments were prepared following the methods in Singer et al. (2000) with adaptations from Barron and Ka’aihue (2003). WAF was made using the ratio of 1: 25 (v/v), fuel to filtered seawater (FSW) following the methods of Brown et al. (2017). Ratios for chemically dispersed treatments were 1: 100 (v/v), fuel to FSW and 1: 20 (v/v) dispersant to fuel. Dispersant only treatments were made using ratios for CEWAF, substituting the fuel component with FSW. Mixes were made in 5 L or 10 L glass aspirator bottles using a magnetic stirrer to achieve a vortex of approximately 20% in the FSW before the addition of test media. The same mixing energy was used to prepare all WAFs for enhanced reproducibility and comparability of results (Barron and Ka’aihue, 2003). Mixes were stirred in darkness to prevent bacterial growth for 18 h with an additional settling time of 6 h at 0 plus or minus 1 oC. A dilution series of four concentrations were made from the full strength aqueous phase of each mix using serial dilution. WAF test concentrations were 100%, 50%, 20% and 10% while CEWAF concentrations were 10%, 5%, 1% and 0.1%. These concentrations were chosen in order to quantify the mortality curve and allow statistical calculation of LC50 values. To facilitate comparisons of dispersant toxicity in the presence and absence of fuel, dispersant only test concentrations reflected those of CEWAF treatments. WAF was sealed in airtight glass bottles stored at 0 plus or minus 1 oC for a maximum of 3 h before use. Fresh test solutions were prepared every four days to ensure consistent water quality and replace hydrocarbons that adsorbed or evaporated into the atmosphere. Each test concentration was represented by five replicates with five FSW control beakers, with approximately 10 S.palludinoides individuals per replicate. The healthiest and most active individuals were chosen. Beakers were filled to 200 ml and were left open to allow the natural evaporation of lighter monoaromatic hydrocarbon components that would occur during a real spill. Animals were not fed during experiments to prevent hydrocarbons being ingested, thereby introducing an additional exposure pathway. Experiments ran for a total of 35 d exposure duration for WAF and CEWAF experiments and 15 d for dispersant only experiments. Experiments were run in cold temperature-controlled cabinets set at a temperature of 0 plus or minus 1 oC, fluorescent lights in the cabinets were set to a light regime of 18 h light, 6 h darkness, following the methods in Brown et al. (2017) to reflect Antarctic summer environmental conditions. Lethal and sublethal observations were made at test times of: 24 h, 48 h, 96 h, 7 d, 8 d, 10 d and 12 d, 14 d, 16 d, 20 d, 21 d, 28 d and 35 d for SAB + Ardrox 6120 experiments and 24 h, 48 h, 96 h, 7 d, 8 d, 10 d and 12 d, 14 d, 15 d for Ardrox 6120 only experiments. The health status of each individual was classified as per the criteria listed below: - Attached to the vial with horns in or out - Unattached (often upside down), horns out, will reattach if flipped over - Not attached but if touched, will retract - Closed but attached and out of water - Operculum closed - Dead, operculum open a little (muscles no longer working), if touched, operculum will not move and tissues might disintegrate Dead animals were removed and preserved in 80% ethanol at each observation period. In order to simulate a repeated pulse pollutant, 90 to 100% of the test solution volume of each beaker was renewed with freshly made test concentrations every four days to replenish hydrocarbons lost through evaporation and adsorption and ensure consistent water quality. Beakers were topped up to 200 ml between water changes with deionised water to maintain water quality parameters. Duplicate 25 ml aliquots of test concentrations were taken at the beginning and end of each experiment in addition to pre and post water change samples. Samples were immediately extracted with 0.7 μm of dichloromethane spiked with an internal standard of BrC20 (1-bromoeicosane) and cyclooctane. Samples were analysed using Gas Chromatography with Flame Ionisation Detection (GC-FID) and mass spectrometry (GC-MS). Brown, K.E., King, C.K., Harrison, P.L., 2017. Lethal and behavioural impacts of diesel and fuel oil on the Antarctic amphipod Paramoera walkeri. Environmental Toxicology and Chemistry. Animal collection, 2013 experiments: animals sourced from AAD aquarium, collected in previous seasons. Animal collection, 2014 experiments: January and February 2014 Experiments were conducted at the Marine Research Facility at the Australian Antarctic Division in Kingston, Tasmania. Experiments using SAB fuel and the fuel dispersant Ardrox 6120 were conducted in August and September 2013, with additional experiments conducted in May 2014 using Ardrox 6120 only.
This metadata record contains the results from bioassays conducted to show the response of the common Antarctic amphipod, Paramoera walkeri to contamination from combinations of Special Antarctic Blend (SAB) diesel, Marine Gas Oil (MGO) and Intermediate Fuel Oil (IFO 180), chemically dispersed with fuel dispersants Ardrox 6120 and Slickgone NS. Fuel only water accommodated fractions (WAF), chemically enhanced water accommodated fractions (CEWAF) and dispersant only treatments were prepared following the methods in Singer et al. (2000) with adaptations from Barron and Ka’aihue (2003). WAF was made using the ratio of 1: 25 (v/v), fuel to filtered seawater (FSW) following the methods of Brown et al. (in prep). Ratios for chemically dispersed treatments were 1: 100 (v/v), fuel to FSW and 1: 20 (v/v) dispersant to fuel. Dispersant only treatments were made using ratios for CEWAF, substituting the fuel component with FSW. Mixes were made in 5 L or 10 L glass aspirator bottles using a magnetic stirrer to achieve a vortex of 20-25% in the FSW before the addition of test media. The same mixing energy was used to prepare all WAFs for enhanced reproducibility and comparability of results (Barron and Ka’aihue, 2003). Mixes were stirred in darkness to prevent bacterial growth for 42 h with an additional settling time of 6 h at 0 plus or minus 1 oC. Extended stirring times were used following the recommendations determined as part of the hydrocarbon chemistry component of this project (Kotzakoulakis, unpublished data). A dilution series of four concentrations were made from the full strength aqueous phase of each mix using serial dilution. WAF test concentrations were 100%, 50%, 20% and 10% while CEWAF concentrations were 10%, 5%, 1% and 0.1%. These concentrations were chosen in order to quantify the mortality curve and allow statistical calculation of LC50 values. To facilitate comparisons of dispersant toxicity in the presence and absence of fuel, dispersant only test concentrations reflected those of CEWAF treatments. WAF was sealed in airtight glass bottles stored at 0 plus or minus 1 oC for a maximum of 3 h before use. Fresh test solutions were prepared every four days to ensure consistent water quality and replace hydrocarbons that adsorbed or evaporated into the atmosphere. Each test concentration was represented by five replicates with five FSW control beakers, with 10 P. walkeri individuals per replicate. Only healthy and active individuals were chosen with a size range of 7.9 plus or minus 0.7 mm for adults and 2.5 plus or minus 0.2 for juveniles measured from the base of the antennae to the widest part of the dorsal curve. Larger individuals and brooding females were not used to avoid unrelated deaths related to age or reproductive state (Sagar, 1980). Beakers were filled to 200 ml and were left open to allow the natural evaporation of lighter monoaromatic hydrocarbon components that would occur during a real spill. A small square of plankton mesh was placed in each jar to provide a substratum to reduce the stress of laboratory conditions and to help to stem cannibalism. Animals were not fed during experiments to avoid hydrocarbons adsorbed onto food pellets being ingested by the amphipods, thereby introducing an additional exposure pathway. Experiments ran for a total of 12 d exposure duration. Experiments were run in cold temperature-controlled cabinets maintained at a temperature of 0 plus or minus 1 oC, fluorescent lights in the cabinets were set to a light regime of 18 h light, 6 h darkness, following the methods in Brown et al. (2017) to reflect Antarctic summer environmental conditions. Lethal and sublethal observations were made at standard ecotoxicology test times of 24 h, 48 h, 96 h, 7 d, 10 d and 12 d, with an additional observation at 8 d coinciding with one of the 4-day water changes. The health status of each individual was classified on a scale of one to four; one showing no effect up to four being mortality. Mortality was determined by a lack of movement and response to stimuli, particularly in the gills. Dead animals were removed and preserved in 80% ethanol at each observation period. Missing amphipods that may have been cannibalised were included in mortality counts as they were likely to have been moribund or already dead when eaten. In order to simulate a repeated pulse pollutant, 90 to 100% of the test solution volume of each beaker was renewed with freshly made test concentrations every four days to replenish hydrocarbons lost through evaporation and adsorption and ensure consistent water quality. Beakers were topped up to 200 ml between water changes with deionised water to maintain water quality parameters. Duplicate 25 ml aliquots of test concentrations were taken at the beginning and end of each experiment in addition to pre and post water change samples. Samples were immediately extracted with 0.7 μm of dichloromethane spiked with an internal standard of BrC20 (1-bromoeicosane) and cyclooctane. Samples were analysed using Gas Chromatography with Flame Ionisation Detection (GC-FID) and mass spectrometry (GC-MS). To determine actual exposure concentrations, four day measured TPH values were used to create a continuous exposure and evaporation profile over the 12 d test period following the methods outlined in Payne et al. (2014) and Brown et al. (2017).
Metadata record for data from ASAC Project 1005 Metal and organic contaminants in marine invertebrates from Antarctica, field study of their concentrations, laboratory study of their toxicities. See the link below for public details on this project. Data from this project are now unrecoverable. Several publications arising from the work are attached to this metadata record, and are available to AAD staff only. Taken from the referenced publications: Bioaccumulation of Cd, Pb, Cu and Zn in the Antarctic gammaridean amphipod Paramoera walkeri was investigated at Casey station. The main goals were to provide information on accumulation strategies of the organisms tested and to verify toxicokinetic models as a predictive tool. The organisms accumulated metals upon exposure and it was possible to estimate significant model parameters of two compartment and hyperbolic models. These models were successfully verified in a second toxicokinetic study. However, the application of hyperbolic models appears to be more promising as a predictive tool for metals in amphipods compared to compartment models, which have failed to adequately predict metal accumulation in experiments with increasing external exposures in previous studies. The following kinetic bioconcentration factors (BCFs) for the theoretical equilibrium were determined: 150-630 (Cd), 1600-7000 (Pb), 1700-3800 (Cu) and 670-2400 (Zn). We find decreasing BCFs with increasing external metal dosing but similar results for treatments with and without natural UV radiation and for the combined effect of different exposure regimes (single versus multiple metal exposure) and/or the amphipod collective involved (Beall versus Denison Island). A tentative estimation showed the following sequence if sensitivity of P. walkeri to an increase of soluble metal exposure: 0.2-3.0 micrograms Cd per litre, 0.12-0.25 micrograms Pb per litre, 0.9-3.0 micrograms Cu per litre and 9-26 micrograms Zn per litre. Thus, the amphipod investigated proved to be more sensitive as biomonitor compared to gammarids from German coastal waters (with the exception of Cd) and to copepods from the Weddell Sea inferred from literature data. ####### This study provides information on LC50 toxicity tests and bioaccumulation of heavy metals in the nearshore Antarctic gammarid, Paramoera walkeri. The 4 day LC50 values were 970 micrograms per litre for copper and 670 micrograms per litre for cadmium. Net uptake rates and bioconcentration factors of these elements were determined under laboratory conditions. After 12 days of exposure to 30 micrograms per litre, the net uptake rates were 5.2 and 0.78 micrograms per gram per day and the bioconcentration factors were 2080 and 311 for copper and cadmium respectively. The body concentrations of copper were significantly correlated with the concentrations of this element in the water. Accumulation of copper and cadmium continued for the entire exposure suggesting that heavy metals concentrations were not regulated to constant concentrations in the body. Using literature data about two compartments (water-animal) first-order kinetic models, a very good agreement was found between body concentrations observed after exposure and model predicted. Exposure of P. walkeri to mixtures of copper and cadmium showed that accumulation of these elements can be assessed by addition of results obtained from single exposure, with only a small degree of uncertainty. The study provides information on the sensitivity of one Antarctic species towards contaminants, and the results were compared with data of similar species from lower latitudes. An important finding is that sensitivity to toxic chemicals and toxicokinetic parameters in the species investigated are comparable with those of non-polar species. The characteristics of bioaccumulation demonstrate that P. walkeri is a circumpolar species with the potential to be a standard biological indicator for use in monitoring programmes of Antarctic nearshore ecosystems. the use of model prediction provide further support to utilise these organisms for biomonitoring. ####### Heavy-metal concentrations were determined in tissues of different species of benthic invertebrates collected in the Casey region where an old waste-disposal tip site is a source of contamination. the species studied included the bivalve Laternula elliptica, starfish Notasterias armata, heart urchins Abatus nimrodi and A. ingens and gammaridean amphipod Paramoera walkeri. The specimens were collected at both reference and contaminated locations where lead was the priority element and copper was the next most important in terms of increased concentrations. The strong association between a gradient of contamination and concentrations in all species tested indicated that they are reflecting well the environmental changes, and that they appear as appropriate biological indicators of heavy-metal contamination. Aspects of the biology of species with different functional roles in the marine ecosystem are discussed in relation to their suitability for wider use in Antarctic monitoring programmes. For example, in terms of heavy-metal bioaccumulation, the bivalve appears as the most sensitive species to detect contamination; the starfish provides information on the transfer of metals through the food web while the heart urchin and gammarid gave indications of the spatial and temporal patterns of the environmental contamination. The information gathered about processes of contaminant uptake and partitioning among different tissues and species could be used in later studies to investigate the behaviour and the source of contaminants.
Metadata record for data from AAS (ASAC) Project 2933. See the child records for access to the datasets. Public While it is generally thought that Antarctic organisms are highly sensitive to pollution, there is little data to support or disprove this. Such data is essential if realistic environmental guidelines, which take into account unique physical, biological and chemical characteristics of the Antarctic environment, are to be developed. Factors that modify bioavailability, and the effects of common contaminants on a range of Antarctic organisms from micro-algae to macro-invertebrates will be examined. Risk assessment techniques developed will provide the scientific basis for prioritising contaminated site remediation activities in marine environments, and will contribute to the development of guidelines specific to Antarctica. Project objectives: 1. Develop and use toxicity tests to characterise the responses of a range of Antarctic marine invertebrates, micro- and macro-algae to common inorganic and organic contaminants. 2. To examine factors controlling bioavailability and the influence of physical, chemical and biological properties unique to the Antarctic environment on the bioavailability and toxicity of contaminants to biota. 3. To compare the response of Antarctic biota to analogous species in Arctic, temperate and tropical environments in order to determine the applicability of using toxicity data and environmental guidelines developed in other regions of the world for use in the Antarctic. 4. Develop a suite of standard bioassay techniques using Antarctic species to assess the toxicity of mixtures of contaminants (aqueous and sediment-bound) including tip leachates, sewage effluents and contaminated sediments. 5. To establish risk assessment models to predict the potential hazards associated with contaminated sites in Antarctica to marine biota, and to develop Water and Sediment Quality Guidelines for Antarctica to set as targets for the remediation of contaminated marine environments. Taken from the 2008-2009 Progress Report: Progress against objectives: Due to logistical constraints, only a short field season (5 weeks) was conducted at Casey in 2008/09 and no berths were allocated solely to this project. A team of 6 scientists worked together on an intensive marine sampling program under TRENZ (AAS project 2948, CI Stark) in support of 5 different AAS projects, including this one. The lack of adequate personnel dedicated to this project and the limited time that we were allocated on station hindered progress and meant that no experiments on Antarctic organisms were able to be conducted in situ. The airlink was however successfully used to transport marine invertebrates collected at Casey and held in seawater at 0degC back to Hobart on 3 separate flights. These invertebrates are currently being maintained in the cold water ecotoxicology aquarium facilities at Kingston. Once they are sorted and where possible established in cultures, they will be used in toxicity tests. Progress against specific objects are: 1) Much effort and time has been put towards the husbandry and culture of the collected Antarctic marine invertebrates. Some species are now successfully breeding in the laboratory providing new generations and sensitive juvenile stages of invertebrates to work with in toxicity tests. This culturing capability, if able to be developed, will hugely extend opportunities for carrying out research for this project, by giving us access to live material over the winter months and during summer when berths to or space on station in Antarctica is limited. Toxicity tests using some of the common amphipods and gastropods collected in the 0809 season at Casey will commence shortly at Kingston. 2) Toxicity tests to commence shortly using invertebrates collected in the 0809 season now being maintained in the Ecotoxicology aquarium will focus on interactions and potentially synergistic effects of contaminants along with other environmental stressors including increases in temperature and decreases in salinity associated with predicted environmental changes in response to climate change. 3) A phD candidate has recently started on this project and is currently reviewing all available literature on the response of Antarctic species to contaminants and environmental stressors in comparison to related species from lower latitudes. 4) Invertebrates collected in the 0809 season that are being maintained in the Ecotoxicology aquarium will be screened in toxicity tests to commence shortly. Methods will then be developed using the most suitable and sensitive species to form the basis of standard bioassay procedures that can be used to test mixtures such as sewage effluents and tip leachates in the upcoming season. 5) The establishment of risk assessment models and Environmental Quality Guidelines for Antarctica is a long term goal of this project when data from the first 4 objectives can be synthesised and hence has not yet been addressed. Taken from the 2009-2010 Progress Report: Progress against objectives: Objectives 1 and 2: Metal effects on the behaviour and survival of three marine invertebrate species were investigated during the field season. Two replicate toxicity tests were conducted on the larvae of sea urchin Sterechinus neumayeri where combined effects of metal (copper and cadmium) and temperature (-1, 1 and 3 degrees Celsius) were to be investigated on developmental success. However, due to lower than optimal fertilisation success, both tests were terminated before any meaningful results could be derived. Four tests were conducted on the adult amphipod, Paramorea walkeri. Two replicate tests investigated combined metal (copper and cadmium) and temperature (-1, 1 and 3 degrees Celsius) effects, and two tests investigated the effects of copper, cadmium, lead, zinc and nickel exposure at ambient sea water temperature of -1 degrees Celsius. One test was conducted with the micro-gastropod Skenella paludionoides being exposed to copper, cadmium, lead, zinc and nickel at ambient sea water temperature. The larvae of bivalve Laternula sp. were also investigated as a potential test organism for metal toxicity. Strip spawning was conducted a number of times, however, this technique did not provide adequate levels of fertilisation success and as such, the toxicity tests on larval development were not completed. Objective 3: A phD candidate working on this project is in the process of compiling a review of all available date on the response of Antarctic species to contaminants and environmental stressors in comparison to related species from lower latitudes. This literature review will form a major component of her thesis' first chapter Objective 4: Methods for Standard bioassay procedures were developed using the most suitable and sensitive species, the amphipod Paramoera walkeri and the microgastropod Skenella paludionoides. These standard tests were then used to assess the toxicity of sewage effluent at Davis Station (in conjunction with project 3217). Objective 5: Toxicity tests on sewage effluent were conducted as part of a risk assessment to determine hazards associated with the current discharge. The determined toxicity of the sewage effluent will provide a basis for guideline recommendations on the required level of treatment and on what constitutes an adequate or 'safe' dilution factor for dispersal of the effluent discharge to the near shore marine environment.
Two three-week toxicity tests were completed at Davis station 2009/10 as part of STP project 3217, to provide environmental information in support of an operational infrastructure project to up-grade sewage treatment at Davis (Project 3157). These tests addressed the third specific objective of the STP projects proposal; to determine the toxicity of sewage effluent as the basis for recommendations on the required level of treatment and on what constitutes an adequate or 'safe' dilution factor for dispersal of the effluent discharge to the near shore marine environment. Results from toxicity tests will provide a baseline by which future changes and/or improvements to the effluent discharge can be quantified (in terms of reduced toxicity and impacts to marine biota), if secondary treatment is re-established. The toxicity of the sewage effluent discharge on marine invertebrates local to the Davis coastal area were assessed using standard bioassay protocols developed and tested under AAS #2933 (King). Two key local invertebrate species were used in both tests; the amphipod Paramoera walkeri and the microgastropod Skenella paludionoides. Invertebrates were exposed to varying concentrations of effluent collected from the outfall pipe (test 1) and from a composite made up of samples collected from the main holding tanks around Davis station (test 2). Each test included two controls with 0% effluent (seawater; SW and hyper-saline brine; HSB) and six concentrations of effluent; 3.125%, 6.25%, 12.5%, 25%, 50% and either 68% (test 1) or 63% (test 2). Dilution gradients of sewage effluent were used in tests in order to calculate the dilution of effluent required to cause no observable effects (NOEC), the lowest observable effect (LOEC) and predictable levels of response (e.g. ECX estimates) in test organisms. Organism responses are thereby used to derive protective concentration values for the effluent and the dilution required to have no predicted effects or impact on the community in the receiving marine environment. The behaviour and mortality of the test species were observed on day 1, 2, 4, 7, 10, 14 and 21 of the three week tests. Test 1 (Discharged Effluent): For test 1, effluent was collected directly from the station outfall line at 11am on 15/2/2010. Discharged effluent was collected in a 20 L bucket, placed at the end of the outfall line. Discharge of effluent from the outfall was initiated by pumping from the holding tank of the Sleeping and Medical Quarters / Old Living Quarters by the plumber (communication with the plumber via radio to co-ordinate the time of release from the tank). The effluent was decanted into containers on site and transported back to the laboratory for testing. Test 2 (Composite Effluent): For test 2, a 1 L sample of effluent was collected from each of the Davis station main building holding tanks, using a long handled ladle on 19/2/10. The composite provided an effluent sample that was representative of the waste generated in all activities on station from domestic activities and from work groups. The volumes of each of the building effluents used to make the final composite test effluent were based on estimates of the volume of effluent created in each of the buildings as a proportion of the total stations effluent discharge and were as follows: Building Name; as per map No. 14148,(Building Acronym),volume Summer Accommodation Module,(SAM),416 mL Temporary Accommodation Davis,(TAD),84 mL Operations Building,(OPS),208 mL Sleeping and Medical Quarters / Old Living Quarters,(SMQ/OLQ),834 mL Living Quarters,(LQ),208 mL Meteorology / Science Buildings,(MET/SCI),84 mL Workshop,(WORKSHOP),84 mL Climate Processes and Change,(ASP),84 mL Location of buildings, within the Davis station area are provided in the Davis Buildings and Structures map (Map Catalogue No. 14148), available from the SCAR Map Catalogue at http://data.aad.gov.au/aadc/mapcat/display_map.cfm?map_id=14148. Effluent dilution to test concentrations: Effluent samples in both tests were adjusted to the required test salinity of 32.4 ppt using HSB to match the salinity of the ambient control SW collected off the coast off Davis and used as Control 1 in tests. The HSB salinity adjusted effluent was then diluted with SW to prepare the concentrations series of effluent (as effluent %) to be used as test solutions in toxicity tests. Each test included six concentrations of effluent; 3.125%, 6.25%, 12.5%, 25%, 50% and either 68% (test 1) or 63% (test 2). Controls for both SW and HSB were also required as both were used as diluents in tests. The SW control used seawater from a field aquarium unit* (also the source of SW used to prepare effluent test solutions) and the HSB control was made using milliQ water that was adjusted to the test salinity using HSB. HSB was made by freezing and partially melting control seawater. The melt water is collected as hyper saline brine. *The field aquarium unit contained locally sourced seawater, collected away from known contaminant sources. Water was physically filtered to 1 micron, biofiltered to remove ammonia, UV sterilised and held at a temperature of -0.8 degrees Celsius. Physico-chemical analysis of test effulents: Physico-chemical characteristics of each of the effluent samples including salinity, dissolved, oxygen, temperature and pH were measured immediately on return to the laboratory using a calibrated multi-meter. Further characterisation of effluents, including coliform counts, microbial analysis, organic content and metal concentrations were conducted and are reported in other DAVIS_STP linked data sets. Test species: Two test species were used in both tests; the amphipod Paramoera walkeri and the microgastropod Skenella paludionoides. These test species were chosen based on their abundance and widespread distribution in nearshore environments around Davis Station. Both species were collected by wading and dip netting from the shoreline at Airport Beach, an uncontaminated site located away from the current site of effluent discharge, and for microgastropods, additional sample were obtained from the surface of macro algae collected locally from boats. Both species were housed in control seawater in the field aquarium prior to their use in tests. Collection dates were 8/02/2010 for both species and for microgastropods also 2/02/2010 from Airport Beach (specific dates not known for other sites). Toxicity Test Set up: For each species, each test consisted of 4 replicate 70 mL vials per concentration (including 6 effluent concentrations and 2 controls) containing 60 mL of test solution. 10 individuals were added to each vial at the start of tests (total 320 individuals per species per test). An additional 10 individuals of each species was sampled and preserved in 100% ethanol for genetic analysis, and a further 10 individuals of each test species fixed in 4% formalin for taxonomy and size range analysis prior to the start of each test. Test Conditions and maintenance: Test vials were kept in culture cabinet at 0 degrees Celsius for the duration of the test. In tests with amphipods, a small strip of plastic mesh was added to each vial to provide a substrate and clinging surface for the amphipods. Invertebrates in test vials were fed and average of 0.036 g of Sera granumarin (granulated fish food) on day 6 of each week of the test. Test solutions were renewed in vials on day 7 of each week using freshly collected effluent for test 1 and a new freshly collected composite effluent for test 2. Test Duration: Observations of individuals in test were made at 24, 48, 96 hr, 7, 10, 14 and 21 days for each of the three week tests. This test duration was chosen based on its relevance to the rate of response of both species to effluent exposure. Individuals were scored as either alive or dead, and all dead invertebrates were removed immediately after scoring. A range of sublethal behavioural endpoints including activity were also investigated but did not reveal useful trends and were therefore not used in the final analyses. Test 1 commenced on 16/02/2010 and was terminated on 9/03/2010. Test 2 commenced on 19/02/2010 and was terminated on 12/03/2010. Data Analysis: For each species and each test, and at each of the 7 time end points (24, 48, 96 hr, 7, 10, 14 and 21 days), NOEC and LOEC values were determined using Dunnett's multiple comparison test. Probit Analysis or Trimmed Spearman Karber Tests with Abbott's correction (if assumptions of the Probit Analysis were not met) and Linear Interpolation with Bootstraping were also used to determine point estimates including EC1, 5, 10 and 50 values, and 95 % confidence limits (CL). If ECX, NOEC or LOEC values were outside the range of concentrations tested, results are reported as greater than (gt) or less than (lt) the highest or lowest concentration tested. All statistical analyses were done using the software Toxcalc for Excel (TidePool Scientific Software, California, 1992). To determine the relative sensitivity of the 2 test species and the toxicity of the 2 effluent samples, EC50 values from tests were compared using ANOVA and SNK tests. An average EC50, NOEC and LOEC was determined for the effluent overall. To investigate the precision of the average EC50 estimate and the consistency of responses between tests, the coefficient of variation (CV) and 95 % CL of EC50 values among tests was also calculated. Data Files Provided: Three data files are provided with this record. 1. STP ECOTOX physico_chem data.xls This file provides water quality measurements for effluent samples collected in association with the two toxicity tests (Test 1 and Test 2) for the three collection events (day 0, 7 and 14). Parameters measured include: pH, salinity, temperature, oxygen content, immediate oxygen demand (IOD) and FDO (FDO). 2. STPECOTOX Test 1 DISCHARGED.xlsx This file provides toxicity test data (endpoint observations) for test 1, using effluent discharged from the Sleeping and Medical Quarters / Old Living Quarters outfall. 3. STPECOTOX Test 2 COMPOSITE.xlsx This file provides toxicity test data (endpoint observations) for test 2, using a composite effluent collected from holding tanks of all Davis station building.
Metadata record for data from AAS (ASAC) Project 2933. See the child records for access to the datasets. Public While it is generally thought that Antarctic organisms are highly sensitive to pollution, there is little data to support or disprove this. Such data is essential if realistic environmental guidelines, which take into account unique physical, biological and chemical characteristics of the Antarctic environment, are to be developed. Factors that modify bioavailability, and the effects of common contaminants on a range of Antarctic organisms from micro-algae to macro-invertebrates will be examined. Risk assessment techniques developed will provide the scientific basis for prioritising contaminated site remediation activities in marine environments, and will contribute to the development of guidelines specific to Antarctica. Project objectives: 1. Develop and use toxicity tests to characterise the responses of a range of Antarctic marine invertebrates, micro- and macro-algae to common inorganic and organic contaminants. 2. To examine factors controlling bioavailability and the influence of physical, chemical and biological properties unique to the Antarctic environment on the bioavailability and toxicity of contaminants to biota. 3. To compare the response of Antarctic biota to analogous species in Arctic, temperate and tropical environments in order to determine the applicability of using toxicity data and environmental guidelines developed in other regions of the world for use in the Antarctic. 4. Develop a suite of standard bioassay techniques using Antarctic species to assess the toxicity of mixtures of contaminants (aqueous and sediment-bound) including tip leachates, sewage effluents and contaminated sediments. 5. To establish risk assessment models to predict the potential hazards associated with contaminated sites in Antarctica to marine biota, and to develop Water and Sediment Quality Guidelines for Antarctica to set as targets for the remediation of contaminated marine environments. Taken from the 2008-2009 Progress Report: Progress against objectives: Due to logistical constraints, only a short field season (5 weeks) was conducted at Casey in 2008/09 and no berths were allocated solely to this project. A team of 6 scientists worked together on an intensive marine sampling program under TRENZ (AAS project 2948, CI Stark) in support of 5 different AAS projects, including this one. The lack of adequate personnel dedicated to this project and the limited time that we were allocated on station hindered progress and meant that no experiments on Antarctic organisms were able to be conducted in situ. The airlink was however successfully used to transport marine invertebrates collected at Casey and held in seawater at 0degC back to Hobart on 3 separate flights. These invertebrates are currently being maintained in the cold water ecotoxicology aquarium facilities at Kingston. Once they are sorted and where possible established in cultures, they will be used in toxicity tests. Progress against specific objects are: 1) Much effort and time has been put towards the husbandry and culture of the collected Antarctic marine invertebrates. Some species are now successfully breeding in the laboratory providing new generations and sensitive juvenile stages of invertebrates to work with in toxicity tests. This culturing capability, if able to be developed, will hugely extend opportunities for carrying out research for this project, by giving us access to live material over the winter months and during summer when berths to or space on station in Antarctica is limited. Toxicity tests using some of the common amphipods and gastropods collected in the 0809 season at Casey will commence shortly at Kingston. 2) Toxicity tests to commence shortly using invertebrates collected in the 0809 season now being maintained in the Ecotoxicology aquarium will focus on interactions and potentially synergistic effects of contaminants along with other environmental stressors including increases in temperature and decreases in salinity associated with predicted environmental changes in response to climate change. 3) A phD candidate has recently started on this project and is currently reviewing all available literature on the response of Antarctic species to contaminants and environmental stressors in comparison to related species from lower latitudes. 4) Invertebrates collected in the 0809 season that are being maintained in the Ecotoxicology aquarium will be screened in toxicity tests to commence shortly. Methods will then be developed using the most suitable and sensitive species to form the basis of standard bioassay procedures that can be used to test mixtures such as sewage effluents and tip leachates in the upcoming season. 5) The establishment of risk assessment models and Environmental Quality Guidelines for Antarctica is a long term goal of this project when data from the first 4 objectives can be synthesised and hence has not yet been addressed. Taken from the 2009-2010 Progress Report: Progress against objectives: Objectives 1 and 2: Metal effects on the behaviour and survival of three marine invertebrate species were investigated during the field season. Two replicate toxicity tests were conducted on the larvae of sea urchin Sterechinus neumayeri where combined effects of metal (copper and cadmium) and temperature (-1, 1 and 3 degrees Celsius) were to be investigated on developmental success. However, due to lower than optimal fertilisation success, both tests were terminated before any meaningful results could be derived. Four tests were conducted on the adult amphipod, Paramorea walkeri. Two replicate tests investigated combined metal (copper and cadmium) and temperature (-1, 1 and 3 degrees Celsius) effects, and two tests investigated the effects of copper, cadmium, lead, zinc and nickel exposure at ambient sea water temperature of -1 degrees Celsius. One test was conducted with the micro-gastropod Skenella paludionoides being exposed to copper, cadmium, lead, zinc and nickel at ambient sea water temperature. The larvae of bivalve Laternula sp. were also investigated as a potential test organism for metal toxicity. Strip spawning was conducted a number of times, however, this technique did not provide adequate levels of fertilisation success and as such, the toxicity tests on larval development were not completed. Objective 3: A phD candidate working on this project is in the process of compiling a review of all available date on the response of Antarctic species to contaminants and environmental stressors in comparison to related species from lower latitudes. This literature review will form a major component of her thesis' first chapter Objective 4: Methods for Standard bioassay procedures were developed using the most suitable and sensitive species, the amphipod Paramoera walkeri and the microgastropod Skenella paludionoides. These standard tests were then used to assess the toxicity of sewage effluent at Davis Station (in conjunction with project 3217). Objective 5: Toxicity tests on sewage effluent were conducted as part of a risk assessment to determine hazards associated with the current discharge. The determined toxicity of the sewage effluent will provide a basis for guideline recommendations on the required level of treatment and on what constitutes an adequate or 'safe' dilution factor for dispersal of the effluent discharge to the near shore marine environment.
The heavy metal content of whole Paramoera walkeri (Eusiridae, Amphipoda) were measured from specimens collected and deployed in experimental mesocosms around Casey station during the summer of 2003/04. Data are the parts per million (ppm) concentrations of 45 heavy metals measured via acid digestion and ICP-MS analysis. P.walkeri were collected from an intertidal area on the northern side of O'Brien Bay and deployed in mesocosms (perforated sample jars housed within perforated 20 litre food buckets) suspended approximately three metres below the sea ice at four sites; two potentially impacted sites in Brown Bay and two control sites, O'Brien Bay and McGrady Cove. The experiment was run on three occasions during the summer each lasting two weeks. These data were collected as part of ASAC project 2201 (ASAC_2201 - Natural variability and human induced change in Antarctic nearshore marine benthic communities). See also other metadata records by Glenn Johnstone for related information.