General description: The associated file contains sediment pigment data from the antFOCE project 4127. Units: all pigment data in ug/g, 0 = below detection limit of HPLC. Sample collection details: At the start and end of the antFOCE experiment, four sediment core samples were taken from inside and outside each chamber or open plot by divers. The top 1 cm of the cores was then removed and placed in the dark, first at -20ºC for 2 hours, then at -80ºC until analysis at the Australian Antarctic division. Pigment analysis Frozen samples were transported under liquid N2 to a freeze drier (Dynavac, model FD-5), in pre-chilled flasks with a small amount of liquid N2 added. Custom made plumbing fitted to the freeze drier enabled samples to be purged with N2 to prevent photo-oxidation up until solvent extraction. Prior to pigment extraction five 2 g stainless steel ball bearings were added to homogenise the freeze dried sediment. The samples were bead beaten for 1 minute (Biospec products). Subsamples (~0.05 g) were immediately transferred to cryotubes with 700 µl of dimethylformamide (DMF) for two hours. Samples were kept at -80ºC and under a safe light (IFORD 902) at all times. All pigment concentrations are standardised to sediment weight. Pigments were extracted with dimethylformamide (DMF 700 µl) over a two hour period at -20ºC. Zirconia beads, and 100 µl of Apo 8 and an internal standard were added to each sub-sample. After a two hour extraction, sub-samples were bead beaten for 20 seconds and then placed in a centrifuge with filter cartridge inserts for 14 minutes at 2500 rpm at -9ºC to separate the solvent from the sediment. The supernatant was transferred into to a vial and placed in a precooled rpHPLC autosampler. The rpHPLC system used is described in Hodgson et al. (1997). Pigment detection was at 435, 470 and 665 nm for all chlorophylls and carotenoids, with spectra from 300–700 nm being collected every 0.2 seconds. Pigment identification was carried out using a combination of rpHPLC and normal phase HPLC retention times, light absorbance spectra and reference standards (see Hodgson et al., 1997). These techniques assisted in the accurate identification of pigments and their derivatives to a molecular level and enabled several pigment derivatives to be analysed. The HPLC was previously calibrated with authentic standards and protocols outlined in SCOR (1988). Data set headers: (A)Treatment: Example code 4127_SOP7_6-1-15_PlotB_R1, = prodject code_Standard Operating Procedure(SOP) used to collect samples(see antFOCE parent file)_ Date_Chamber/plot(A,B,C,D)_replicate core within Chamber/plot(1,2,3) (B) BB carot= BB caroten, type of pigment detected by HPLC. See Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more details. (C) Chl c1 = Chlorophyll derivatives see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (D) Chl c2 = Chlorophyll derivatives see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (E) Chl c3 = Chlorophyll derivative see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (F) Chla = Chlorophyll a see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (G) Ddx =Diadinoxanthin see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information (H) dtx = Diatoxanthin pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information (I) epi = Chlorophyll epimer pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (j) Fuc = Fucoxanthin pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (k) Gyro2 = Gyroxanthin pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (L) Pras = Prasanthin pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (m) Zea = Zeaxanthin pigment. see Wright, S.W., Jeffrey, S.W. and Mantoura, R.F.C. eds., 2005. Phytoplankton pigments in oceanography: guidelines to modern methods. Unesco Pub for more information. (n) Date = Samples taken at the start of antFOCE experiment or at the end (o) chamber = The antFOCE chamber (A,B,C,D) (p) Treatment = The associated pH level in chambers (Acidified ~7.8, Control ~8.2) (Q) Position = Samples were taken within chambers and outside chambers (outside, inside) (r) rep= Subsamples were taken within each chamber/position (R1=replicate one, R1-R4) Spatial coordinates: 66.311500 S, 110.514216 E Dates: between 1/12/2014 and 1/3/2015 Timezone:UTC+11

The impact of freeze-thaw cycling on a ZVI and inert medium was assessed using duplicated Darcy boxes subjected to 42 freeze-thaw cycles. This dataset consists of particle sizing during the decommissioning process of the experiment. Two custom built Perspex Darcy boxes of bed dimensions: length 362 mm, width 60 mm and height 194 mm were filled with a mixture of 5 wt% Peerless iron (Peerless Metal Powders and Abrasive, cast iron aggregate 8-50 US sieve) and 95 wt% glass ballotini ground glass (Potters Industries Inc. 25-40 US sieve). This ratio of media was selected to ensure that most aqueous contaminant measurements were above the analytical limit of quantification (LOQ) for feed solutions at a realistic maximum Antarctic metal contaminant concentration at a realistic field water flow rate. All solutions were pumped into and out of the Darcy boxes using peristaltic pumps and acid washed Masterflex FDA vitron tubing. Dry media was weighed in 1 kg batches and homogenised by shaking and turning end over end in a ziplock bag for 1 minute. To ensure that the media was always saturated, known amounts of Milli-Q water followed by the homogenised media were added to each box in approximately 1 cm layers. 20 mm of space was left at the top of the boxes to allow for frost heave and other particle rearrangement processes. On completion of freeze-thaw cycling and solution flow (refer to Statham 2014), an additional series of assessments was conducted. The media from between the entry weir and the first sample port was removed in five approximately 400 g samples of increasing depth. This procedure was repeated between the last sample port and the exit weir. These samples were left to dry in a fume cabinet before duplicated particle sizing using a Endcotts minor sieve shaker.

This metadata record was created in error and a DOI assigned to it before the error was noticed. The correct metadata record is available here: https://data.aad.gov.au/metadata/records/AAS_4015_Krill_Gonad_Transcriptome with the DOI doi:10.26179/5cd3c8fec9ad8.

Microscopy imaging of live Antarctic krill using a Leica M205C dissecting stereo-microscope with a Leica DFC 450 camera and Leica LAS V4.0 software. Krill were held in a custom made 'krill trap', details provided in manuscript in section eight of this form. The data are available as a single video file. These data are part of Australian Antarctic Science (AAS) projects 4037 and 4050. Project 4037 - Experimental krill biology: Response of krill to environmental change The experimental krill research project is designed to focus on obtaining life history information of use in managing the krill fishery - the largest Antarctic fishery. In particular, the project will concentrate on studies into impacts of climate change on key aspects of krill biology and ecology. Project 4050 - Assessing change in krill distribution and abundance in Eastern Antarctica Antarctic krill is the key species of the Southern Ocean ecosystem. Its fishery is rapidly expanding and it is vulnerable to changes in climate. Australia has over a decade of krill abundance and distribution data collected off Eastern Antarctica. This project will analyse these datasets and investigate if krill abundance and distribution has altered over time. The results are important for the future management of the fishery, as well as understanding broader ecological consequences of change in this important species.

This video is supplementary data for the publication entitled 'Internal physiology of live krill revealed using new aquaria techniques and mixed optical microscopy and optical coherence tomography (OCT) imaging techniques'. The video is high resolution microscopy video of a live krill captured in the krill containment trap placed within the water bath. File size: 1.8 GB, 32 s duration. The optical microscopy was carried out using a Leica M205C dissecting stereomicroscope with a Leica DFC 450 camera and Leica LAS V4.0 software to collect high-resolution video. The experimental krill research project is designed to focus on obtaining life history information of use in managing the krill fishery - the largest Antarctic fishery. In particular, the project will concentrate on studies into impacts of climate change on key aspects of krill biology and ecology.

Trace metal concentrations are reported in micrograms per gram of sediment in core C012-PC05 (64⁰ 40.517’ S, 119⁰ 18.072’ E, water depth 3104 m). Each sediment sample (100-200mg) was ground using a pestle and mortar and digested following an initial oxidation step (1:1 mixture of H2O2 and HNO3 acid) and open vessel acid on a 150 degree C hotplate using 2:5:1 mixture of concentrated distilled HCl, HNO3 and Baseline Seastar HF acid. After converting the digested sample to nitric acid, an additional oxidation step was performed with 1:1 mixture of concentrated distilled HNO3 and Baseline Seastar HClO4 acid. A 10% aliquot of the final digestion was sub-sampled for trace metal analyses. Trace metal concentrations were determined by external calibration using an ELEMENT 2 sector field ICP-MS from Thermo Fisher Scientific (Bremen, Germany) at Central Science Laboratory (University of Tasmania). The following elements were analysed in either low (LR) or medium resolution (MR): Sr88(LR), Y89(LR), Mo95(LR), Ag107(LR), Cd111(LR), Cs133(LR), Ba137(LR), Nd146(LR), Tm169(LR), Yb171(LR), Tl205(LR), Pb208(LR), Th232(LR), U238(LR), Na23(MR), Mg24(MR), Al27(MR), P31(MR), S32(MR), Ca42(MR), Sc45(MR), Ti47(MR), V51(MR), Cr52(MR), Mn55(MR), Fe56(MR), Co59(MR), Ni60(MR), Cu63(MR), Zn66(MR).

Metadata record for data from ASAC Project 2547 See the link below for public details on this project. Pue (greater than 90% as determined by SDS-PAGE) samples of nitrate reductase have been isolated from the Antarctic bacterium, Shewanella gelidimarina (ACAM 456T; Accession number U85907 (16S rDNA)). The protein is ~90 kDa (similar to nitrate reductase enzymes characterised from alternate bacteria) and stains positive in an in-situ nitrate reduction (native) assay technique. The protein may be N-terminal blocked, although further sequencing experiments are required to confirm this. This work is based upon phenotyped Antarctic bacteria (S. gelidimarina; S.frigidimarina) that was collected during other ASAC projects. (Refer: Psychrophilic Bacteria from Antarctic Sea-ice and Phospholipids of Antarctic sea ice algal communities new sources of PUFA [ASAC_708] and Biodiversity and ecophysiology of Antarctic sea-ice bacteria [ASAC_1012]). The download file contains 4 scientific papers produced from this work - one of these papers also contains a large set of accession numbers for data stored at GenBank.

This metadata record contains the results from bioassays conducted to show the response of larval Antarctic Sterechinus neumayeri sea urchins to contamination from combinations if IFO 180 fuel and the fuel dispersant Slickgone NS. AAS project 4142. Experiments used an intermediate grade Fuel Oil (IFO 180) and an internationally approved fuel dispersant, Slickgone NS, produced by Dasic International LTD. Treatments included a physically dispersed treatment of IFO 180 only, a chemically dispersed treatment of IFO 180 treated with Slickgone NS and a Slickgone NS only treatment to determine the toxicity of the dispersant. Treatments were experimentally mixed using a magnetic stirrer to combine treatment substances and filtered seawater (FSW) in temperature-controlled cabinets at 0oC to create a Water Accommodated Fraction (WAF). WAFs were produced in 2 L and 5 L glass aspirator bottles following the methods of Singer, Aurand et al. (2000) with adaptations by Barron and Ka'aihue (2003) and Kostzakoulakis (chemistry section, project 4142) stirring for 42 h with a settling time of 6 h. WAF treatments used concentrations of 100%, 50%, 20% and 10%, CEWAF and dispersant only treatments used concentrations of 10%, 5%, 1% and 0.1%. Toxicity tests were conducted in temperature-controlled cabinets at 0 oC using uncapped, forty-millilitre glass headspace vials, each containing 15.5 ml of test solution and 0.5 ml of embryo suspension. Fertilisation methods followed standard procedures for Sterechinus neumayeri. Two tests were conducted to determine the effect of a single pollution event (test 1) compared with a recurring repeated pulse pollution event (test 2). Test 1 required no water changes, while test 2 required renewal of the test treatments on a 4-day basis. Three endpoints were used, un-hatched blastula (48 h to 48.5 h) to represent the embryonic phase, gastrula (10 d) and 4-armed pluteus (16 d to 18 d). At the termination of each endpoint, 1 ml of 10% buffered formalin was added to each relevant vial and recapped. At the conclusion of the experiments, preserved embryos were observed under a dissecting microscope to determine the number of normal, abnormal and unfertilised embryos relative to controls. Samples for analysis of total petroleum hydrocarbon content were taken throughout the 2 experiments to determine the actual concentrations to which embryos and larvae were exposed. The measured concentrations were integrated following the methods of Payne et al. (2014) to obtain a profile of hydrocarbon content over each test period. Two spreadsheets are included in this metadata record detailing survival data and results of hydrocarbon analysis. The survival data file includes test condition details on the first tab, with data for tests 1 and 2 on the second and third tabs. Test treatment and concentration are listed on the left of each data block and count categories are defined in the top left panel. Development stage, date preserved and age of organism is defined for each data block, representing the three endpoints included in the experiments: unhatched blastula, gastrula and 4-armed pluteus. The hydrocarbon analysis, TPH (total petroleum hydrocarbon) file details chemical analysis results produced by K. Kotzakoulais at Macquarie University as part of project 4142. Row terminology explanations are as follows: TPH metadata Test name- indicates the tested species Exp number-indicates whether the data belongs to test 1 or test 2 Water change- details the identification of the sample in relation to the 4-day water change regime. Start samples represent the beginning of the experiment. Pre samples are taken at the end of the corresponding 4-day period, before the water is changed. 'Post' samples are taken of newly made test solutions. The chronological order of sampling is therefore: Start, pre4d, post4d, pre8d, post 8d etc. Only 'pre' samples were taken for test 1, as there were no water changes. less than C9 - greater than C28- Hydrocarbon content of samples was broken down into four compound size classes detailed for each analysis. Contamination- contamination was detected in samples, the source of contamination remains unclear, however it was established that contamination occurred during the sampling process and therefore did not come into contact with organisms. Contamination was therefore excluded from calculations. The hydrocarbon content of 0.1% dilutions was unable to be reliably analysed due to accuracy of the equipment and interfering contamination. Control data indicates spot checks to confirm the presence or absence of fuel. Very small amounts of hydrocarbons were detected as lighter fuel components evaporated and dissolved into control water within the cabinet. These very small amounts are negligible. Abnormality metadata Tab 1 details test conditions Tab 2 'Test 1' includes the data for test 1 Tab 2 'Test 2' includes the data for test 2. All observational categories are defined within the spreadsheet.

This dataset is a document describing the Pelagic Tunicates of the Southern Ocean. It lists all the known Southern Ocean species and with illustrated diagrams provides a guide to their taxonomic identification. The document is available for download as a pdf from the provided URL.

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).