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

This metadata record will contain the results of bioassays conducted to characterise the response of Antarctic near-shore marine zooplankton to hydrocarbon contaminants in Special Antarctic Blend (SAB) diesel fuels commonly used in Antarctica. The results from one summer season (2010-11) are in this record. This was conducted under the AAS Project 3054: Ecological risks from oil products used in Antarctica: characterising hydrocarbon behaviour and assessing toxicity on sensitive early life stages of Antarctic marine invertebrates. Exposure solutions of fuel were experimentally mixed by slow stir of fuel and seawater in temperature control cabinets at -1 degree 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 1:24 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. Ecotoxicological bioassays were conducted at Davis Stations in the 2010/11 summer season using SAB WAF to prepare experimental treatments consisting of WAF dilution series. For each bioassay, treatments consisted of undiluted 100% WAF and dilutions of 10%, 17%, 25% and 50% of WAFs in FSW, to test the toxicity of water accommodated fractions of these three fuels on Antarctic both the zooplankton community and single copepod species. Bioassays were conducted in open vessels (glass jars or beakers) in temperature controlled cabinets. Mortality was observed at endpoints of 24 hrs, 48 hrs, 96 hrs, 7 days, 8 days, 9 days, 10 days, 11days, 12 days, 14 days, 15 days, and 16 days. New WAF solutions were prepared at the 7 day interval 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. Samples of test treatments for chemical analysis of hydrocarbon concentration were taken at each water change. Results of these analyses are not included as delayed progress with HC analyses impacted on quality of samples and these data were not used. This dataset consists of Excel spreadsheets. The file name code for zooplankton bioassays is; Project number_Season_Taxa_Test name Eg AAS_3054_10-11_zooplankton_m1 Project number : AAS_3054 Season : 2010/11 season Taxa: Zooplankton Community Test name: M1 =Multi-species test 1 Bioassay spreadsheets contain the results of bioassays for a species or the zooplankton community. Where replicate tests were conducted, each experiment is on a separate worksheet. The worksheet labelled "Test conditions" shows details of Test name, dates, animal collection details, laboratory holding conditions, details of water accommodated fractions (WAF), and bioassay conditions. The worksheet labelled "Counts" has a table for each of the replicates, arranged into a column for each treatment type. These tables show the number or dead individuals which were found and removed at each of the observation days. The worksheet labelled "Totals" has calculations of total number of individuals (of all species) which were found dead at each observation day in each replicate. It also gives the mean and standard deviation for each of the treatments. Further information on the zooplankton community structure in the 6 samples taken across the summer, based on the community in the toxicity tests and trials, is also included in the spreadsheet "AAS_3054_10-11_zooplankton_CommStructure". Sampling locations were near-shore from Davis Station, Vestfold Hills and from O'Gorman Rocks, southwest of Anchorage Island and northwest of Plough Island.

Experimental Design A six-level, dose-response ocean acidification experiment was run on a natural microbial community from nearshore Antarctica, between 19th November and 7th December 2014. Seawater was collected from approximately 1 km offshore of Davis Station, Antarctica (68◦ 35’ S, 77◦ 58’ E), pre-filtered (200 μm), and transferred into six 650 L tanks (minicosms) located in a temperature-controlled shipping container. Six CO2 levels were achieved by altering the fugacity of carbon dioxide (ƒCO2) within each minicosms. The ƒCO2 was adjusted stepwise to the target concentrations for each minicosm (343, 506, 634, 953, 1140, 1641 μatm) over a five-day period using 0.2 μm filtered seawater enriched with CO2. This acclimation to CO2 was conducted at low light (0.9 ± 0.2 μmol m−2 s−1) so there was low growth of the phytoplankton. Light levels were then increased over a further two days to 90.52 ± 21.45 μmol m−2 on a 19:5 light/dark non-limiting light cycle. After this acclimation period, the microbial community was allowed to grow for 10 days (days 8-18), during which the ƒCO2 levels within each minicosm was adjusted daily to maintain the target ƒCO2 level for each minicosm, and light levels were kept constant. No nutrients were added during the experiment. For a more detailed description of minicosm set-up, lighting and carbonate chemistry see; Davidson, A. T., McKinlay, J., Westwood, K., Thomson, P. G., van den Enden, R., de Salas, M., Wright, S., Johnson, R., and Berry, K.:Enhanced CO2 concentrations change the structure of Antarctic marine microbial communities, Mar. Ecol. Prog. Ser., 552, 93-113, 2016. Deppeler, S. L., Petrou, K., Westwood, K., Pearce, I., Pascoe, P., Schulz, K. G., and Davidson, A. T. Ocean acidification effects on productivity in a coastal Antarctic marine microbial community, Biogeosciences, 15(1), 2018. Sample Collection Samples of 40-400 L were collected and sequentially size-fractionated filtered onto 293 mm biomass filters with 3.0 and 0.1 μm pore-sized polyethersulfone membrane filters (Pall XE20206 Disc 3.0 μm Versapor 293 mm and 656552 Disc 0.1 μm Supor 293 mm) using the design of the Global Ocean Sampling expedition (Rusch et al., 2007). Samples were collected on days 0 (immediately after seawater collection), 12 (mid-exponential growth) and 18 (end of experiment). On day 0, 400 L of seawater was collected from the reservoir tank (pre-filtered 200 μm), from which all the minicosms were filled, to allow characterisation of the initial community. This sample was collected from the reservoir, and not the minicosms, due to the large volume needed to collect sufficient microbial biomass on the filters. On day 12 and 18, 40 L was collected from each minicosm for filtration. The later samples were of a smaller volume due to the increase in biomass in the minicosms during the experiment, meaning less volume of water was required to gain sufficient material on the filters to perform molecular analysis. The filter membranes containing the concentrated microbial biomass were stored in 15 mL of storage buffer, flash frozen in liquid nitrogen and stored at - 80◦C. The storage buffer was freshly prepared on each sampling day with a mixture of 2.5 mM EGTA, 2.5 mM EDTA, 0.1 mM Tris-EDTA, RNA Later (0.5x house prepared), 1 mM PMSF and Protease Inhibitor Cocktail VI (Ng et al., 2010). Between samples the filtration apparatus was sequentially washed with 2 x 25 L 0.1 M NaOH, 2 x 25 L 0.07% Ca(OCl)2 and 2 x 25 L fresh water. All samples were stored and transported at -80◦C to the Australian Antarctic Division, Hobart, Australia for DNA extraction. DNA Extraction and Sequencing The DNA was extracted from half of each filter (3.0 and 0.1 μatm per sample) via the method described in Rusch et al. (2007). In short, the filters were cut into small pieces and agitated in a lysozyme and sucrose buffer for 60 minutes and underwent three freeze/thaw cycles in a Proteinase K solution. This was followed by a gentler agitation

Metadata record AAS_4127_antFOCE_SedimentFauna contains all data sets relating to the fauna sampled from marine sediments during the antFOCE experiment, including macrofauna, meiofauna, diatoms, microbes, as well as DNA sequencing conducted on some of these groups and the results of a sediment bioturbation study. Refer to antFOCE report section 4.4 for deployment, sampling and on-station analysis details. https://data.aad.gov.au/metadata/records/AAS_4127_antFOCE_Project4127 Background The antFOCE experimental system was deployed in O'Brien Bay, approximately 5 kilometres south of Casey station, East Antarctica, in the austral summer of 2014/15. Surface and sub-surface (in water below the sea ice) infrastructure allowed controlled manipulation of seawater pH levels (reduced by 0.4 pH units below ambient) in 2 chambers placed on the sea floor over natural benthic communities. Two control chambers (no pH manipulation) and two open plots (no chambers, no pH manipulation) were also sampled to compare to the pH manipulated (acidified) treatment chambers. Details of the antFOCE experiment can be found in the report – "antFOCE 2014/15 – Experimental System, Deployment, Sampling and Analysis". This report and a diagram indicating how the various antFOCE data sets relate to each other are available at: https://data.aad.gov.au/metadata/records/AAS_4127_antFOCE_Project4127

These data contain results from grazing dilution experiments conducted during BROKE-West. Experiments were conducted at 22 locations on the BROKE-West transect. Data are presented in an excel spreadsheet containing sample collection information (longitude, latitude, UTC date and time, depth), experiment details (incubation time, dilution series), experiment results (chlorophyll a, bacterial concentrations, heterotrophic flagellate concentrations, phytoplankton concentrations, microzooplankton concentrations, geometric mean predator density, phytoplankton growth rates, microzooplankton grazing rates for bacteria and phytoplankton, bacterial growth rates). This work was completed as part of ASAC projects 2655 and 2679 (ASAC_2655, ASAC_2679).

This data set was collected from two minicosm experiments conducted at Davis Station, Antarctica. 1. Variance experiment - 2013/14 summer season 2. Ocean acidification experiment - 2014/15 summer season It includes: - description of methods for all data collection and analyses. - environmental data logged throughout the experiment; nutrients, temperature, light climate. - flow cytometry counts; autotrophic cells, heterotrophic nanoflagellates, and prokaryotes. - FlowCam counts; individual phytoplankton species data. - microscopy counts; individual phytoplankton species data.

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

The data describe all zooplankton species abundance values estimated at each rectangular midwater trawl (RMT 1+8) target and routine sampling site collected during the 2021 TEMPO voyage on R/V Investigator. During the TEMPO voyage, zooplankton were collected using the RMT 1+8 plankton net and sorted down to the lowest taxonomic level possible. The supporting data set, in CSV format, feature abundances estimated from RMT+1 samples only. The data file is named allzooplankton.csv. The fields (columns) in the file are: "Site" – sampling site ID for each routine (RXX) and target (TXX) trawl using RMT1+8. "Date" – date of each sample collected (ie. target or routine trawl) [dd/mm/yyyy]. "Latitude" – latitude of each trawl [degrees] (-dd.ddddd). "Longitude" – longitude of each trawl [degrees] (dd.ddddd). "Chla" – Integrated estimate of chlorophyll-a (mg m^-2) averaged at depths between 10 to 200 m, which aligns with the section of the water column where zooplankton were sampled. "Depth" – Bathymetric depth estimated at each sampling station. "MLD_03" – Mixed layer depth (m). Not used in final analysis. "MLD_05" – Mixed layer depth (m). Not used in final analysis. "MLD_N2max" – Mixed layer depth (m) where the N^2 maximum is found, i.e., the pycnocline. "CT_200" – Absolute temperature (°C) averaged at depths between surface and 200 m maximum net sampling depth. Values are derived from in situ CTD measurements at each sampling site. "SA_200" – Absolute salinity (g kg^-1) averaged at depths between surface and 200 m maximum net sampling depth. Values are derived from in situ CTD measurements at each sampling site. "IceMeltDays" – Number of days since sea ice melted calculated from daily passive microwave estimated percent sea ice concentration taken from the National Snow and Ice Data Centre. "m_to_Ice" – Distance (m) estimated from sampling site to edge of ice. "IceConc" – Actual ice concentration value (%). It is the percentage of an area covered in sea ice. The remaining columns are zooplankton species abundances (individuals per 1000 cubic metres) estimated from count data, volume of original sample, split of sample analysed under the microscope and calibration of the flowmeter: "R.gigas" – Rhincalanus gigas, copepod "C.propinquus" – Calanus propinquus, copepod "C.simillimus" – Calanus simillimus, copepod "C.acutus" – Calanoides acutus, copepod "Metridia sp." – Metridia species, copepod "M.gerlachei" – Metridia gerlachei, copepod "Paraeuchaeta sp." – Paraeuchaeta species, copepod "Pleuromamma sp." – Pleuromamma species, copepod "Oncaea" – Oncaea species, copepod "O.antarctica" – Oncaea antarctica, copepod "Neo tonsus" – Neocalanus tonsus, copepod "Neo gracilis" – Neocalanus gracilis, copepod "Stephos longipes" – Stephos longipes, copepod "Ctenocalanus sp." – Ctenocalanus species, copepod "Microcal pygmaeus" – Microcalanus pygmaeus, copepod "Oithona similis" – Oithona similis, copepod "Oithona frigida" – Oithona frigida, copepod "Heterhabdus australis" – Heterorhabdus australis, copepod "Heterhabdus sp." – Heterorhabdus species, copepod "Harpacticoid" – Harpacticoid species, copepod "Haloptilus sp." – Haloptilus species, copepod "Candacia sp." – Candacia species, copepod "Solecithricella sp." – Solecithricella species, copepod "Clausocalanus breviceps" – Clausocalanus breviceps, copepod "Clausocalanus laticeps" – Clausocalanus laticeps, copepod "Clausocalanus sp." – Clausocalanus species, copepod "Aetideops antarctica – Aetideopsis antarctica, copepod "Aetide minor" – Aetideopsis minro, copepod "Euchirella rostromanga" – Euchirella rostromanga, copepod "Heterostylites longicornis" – Heterostylites longicornis, copepod "Gaidius tenuispinus" – Gaidius tenuispinus, copepod "Gammaridean amphipod" – Gammaridean, amphipod "T.gaudichaudii" – Themisto gaudichaudii, amphipod "Parathemisto sp." – Parathemisto species, amphipod "Hyperia antarctica" – Hyperia antarctica, amphipod "Hyperia sp." – Hyperia species, amphipod "Hyperiella dilatata" –

From the abstract of one of the papers: Phytoplankton biomass and speciation were monitored at an inshore site near Davis Station, East Antarctica during three consecutive summer seasons (December-February, 1992-5). Four distinct phytoplankton assemblages were identified in which the dominant species were: Phaeocystis sp., an undescribed Cryptomonas species, Thalassiosira dichotomica, and a mixed assemblage containing Fragilariopsis spp. and Nitzschia spp. Little interannual consistency was found in either the timing of the appearance or disappearance of the various assemblages. Similarly, the seasonal trends in biomass varied dramatically from year to year. Variations in the phytoplankton community can be ascribed, to some extent, to the random variation in a number of factors, including the date of fast ice break out, water column stratification, temperature and salinity, zooplankton grazing and strong winds. Periods of strong wind result in the introduction of offshore or deeper water masses into the shallow inshore environment, where the physical and chemical conditions allow blooms to develop. A number of the papers listed in the reference section are available as pdf's in the download section.