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.

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.

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

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.

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).

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.

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 data set was collected from a ocean acidification minicosm experiment performed at Davis Station, Antarctica during the 2014/15 summer season. It includes: - description of methods for all data collection and analyses. - flow cytometry counts; autotrophic cells, heterotrophic nanoflagellates, and prokaryotes

This dataset contains results of toxicity tests with early life stages of the sea urchin Sterechinus neumayeri as part of 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.' Dataset consists of excel spreadsheets with separate spreadsheets for each test. Test details are outlined on worksheets 'Test conditions' and results of test in worksheet 'Counts'. This metadata record contains the results of toxicity tests conducted to characterise the response of Antarctic nearshore marine invertebrates to hydrocarbon contaminants in fuels commonly used in Antarctica as part of AAS Project 3054. This dataset contains results of toxicity tests conducted at Davis Station in 2010/11 summer season to test the sensitivity of fertilisation and early life stages of the sea urchin Sterechinus neumayeri to fuels in seawater. The three fuel types used 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 controlled 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 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 then settled for 6 h before the water portion was drawn from beneath the fuel. Mature S. neumayeri were collected from the outlet of Ellis Fjord, East Antarctica (68.62°S, 77.99°E) in December and early January 2010/11. Sea urchins were collected from shallow nearshore waters less than 1m deep, placed in 20 L buckets of seawater and transported to Davis station. They were held for 1–2 d in a flow-through aquarium at -1 plus or minus 1°C, with macroalgae from the collection site as a food source, before being used for testing. Seawater for experiments was collected ~20 m from the shoreline north of Davis station (68°34’ S, 77°57’ E). Collected seawater was filtered to 0.45 µm (FSW) and stored in 30 L polyethylene containers at 0°C. Fertilisation and early embryo toxicity tests. Effects of WAFs on fertilisation and on development to the 2 cell stage were determined in static tests in which both eggs and sperm were pre-exposed to SAB, MGO and IFO 180 WAFs, fertilised within treatments and developed to the 2 cell stage (G1, G2, G3). Gamete exposure and fertilisation was done in a temperature controlled room at 0°C. Test vessels were 22 mL borosilicate glass vials with foil lined lids holding 20 mL of test solution. There were 10 vials for each treatment; 5 replicates for fertilisation and 5 replicates for the 2 cell endpoint. To pre-expose eggs, 5 mL of prepared egg solution was added to vials that contained 5 mL of 2, 20 and 100% WAFs and FSW controls, to give final treatment concentrations of 1, 10 and 50% WAF dilutions and FSW controls. Vials were sealed, swirled gently to mix and left standing for 20 min. To pre-expose sperm, pooled sperm were activated by dilution in FSW to the density required for a sperm to egg ratio of 800:1. One µL of sperm solution was added to vials containing 5 mL of FSW and gently mixed. Five mL of this solution was then added to vials containing 5 mL of 2%, 20% and 100% WAFs (final treatments of 1, 10 and 50% WAF dilutions) and FSW controls. The vials were sealed, swirled gently to mix and left for 15 mins. After the gamete exposure period was complete, for each treatment the contents of the sperm vials were added to the egg vials with a final target concentration of ~10 eggs per mL. Vials were sealed and placed into temperature-controlled cabinets set at -1 plus or minus 1°C. Temperature was recorded at 10 min intervals using a data logger (Maxim ibutton) and averaged -1.3 plus or minus 0.5°C. Tests were terminated at 4 h for the fertilisation endpoint, and at 11 h for the 2 cell endpoint by the addition of 1 mL of 2.5% (v/v) buffered glutaraldehyde. Samples were viewed in a Sedgewick Rafter counting cell under a compound microscope at 10 times magnification. Fertilisation was assessed according to the presence or absence of a fertilisation membrane in the first 100 eggs counted, to obtain the percentage of eggs fertilised in each replicate. The 2 cell endpoint was assessed in the first 100 embryos counted, as the percentage of embryos in each replicate with normal first cleavage. Embryonic and larval toxicity tests. Effects of fuel WAFs on embryonic and larval development were tested with 1, 10, and 100% WAFs of SAB, MGO and IFO 180 and FSW control, with 5 replicates per treatment. Eggs and sperm were collected and density of solutions adjusted as described above to obtain the optimal sperm to egg ratio of 800:1. Two semi-static tests (EL1, EL2) were done to test effects of WAFs on embryos and larvae when first exposed as zygotes (eggs fertilised in FSW then exposed to treatments before the first cleavage). To fertilise eggs, sperm were activated by their addition to 10 mL of FSW, and 1 µL of this sperm solution was added to beakers containing 700 mL of egg solution and gently mixed. After two hours, the mixture was stirred with a glass rod to maintain a homogeneous suspension while aliquots were transferred into 100 mL glass vials filled with 80 mL of test treatment, to a final density of ~10 zygotes per mL. Three tests (GL1, GL2, GLP) were done to test effects of WAFs on larval development with exposure commencing as gametes. One mL aliquots of egg mixture were added to vials containing 80 mL of test solution (to a density of ~10 eggs per mL) and left for 20 min. Sperm were activated in 10 mL of FSW and 0.1 mL aliquots added to the vials to fertilise eggs within treatments at a sperm to egg ratio of 800:1. Two exposure regimes were used; continuous semi-static WAF renewal (GL1 and GL2) and a single static pulse of WAF exposure up to the 4 d unhatched blastula stage, followed by post exposure recovery in FSW up to the 21 d pluteus stage (GLP). Vials were left uncovered and placed in a temperature controlled cabinet at -1 plus or minus 1°C with an 18 h light, 6 h dark photoperiod. Tests were under semi-static conditions, with test solutions renewed every 4 d. Water quality data was collected at each water change. Treatment renewals were done by removing and replacing approximately 90% of test solution. Disposable syringes with silicon tubing attached to the nozzle, and with the end of the tubing covered with plankton mesh, were used to withdraw test solution while preventing embryos/larvae from being removed. The vials were then refilled to the 80 mL mark with fresh test solutions. Treatment renewals for tests EL1, EL2 and GL1, GL2 were with freshly made WAFs every 4 d. For the single pulse WAF exposure test (GLP) on the first treatment renewal at 4 d, treatment solutions were removed as described above, and replaced with FSW. All subsequent 4 d renewals for test GLP were with FSW. To maintain the volume and salinity of test treatments a small volume of purified and deionised (Milli-Q) water at -1°C was stirred into the vials to the 80 mL mark every 2 d between water changes. Water quality measurements were made at the start of tests and pre and post treatment renewals. Mean water quality parameter measurements were pH 8.08 plus or minus 0.10, salinity 36.6 plus or minus 0.9‰ and dissolved oxygen 11.1 plus or minus 0.61 mg/L. Temperature was recorded at 10 min intervals using a data logger (Maxim ibutton) and averaged -1.0 plus or minus 1.0°C. In tests where exposure commenced as zygotes, endpoints were the embryonic 4-8 cell (20 h) and unhatched blastula (48 h) stages, and the larval blastula (6–7 d) and gastrula (14–15 d) stages. In tests with exposure commencing as gametes, endpoints were the larval blastula, gastrula and early 4-arm pluteus (21–24 d) stages. At each endpoint a sample was taken from each replicate by drawing an aliquot with a glass pipette and transferring it to a vial, to which 1 mL of 2.5% (v/v) buffered glutaraldehyde was added. Embryo and larvae were viewed in a Sedgewick Rafter counting cell under a compound microscope at 10 times magnification. The first 30 individuals in each sample at the 4-8 cell and unhatched blastula endpoints, and the first 100 individuals at the blastula, gastrula and pluteus endpoints, were assessed for normality. Test EL1 ended at the blastula stage and tests EL2 and GL2 at the gastrula stage as there were insufficient numbers of larvae remaining to continue the test beyond these stages. All remaining larvae were counted at the final endpoint. Chemical analysis of water accommodated fractions Total hydrocarbon content (THC) in WAFs were derived from replicate tests conducted under the same conditions but without test organisms. In these tests at 0°C, the concentrations of freshly made WAFs of each of the three fuels, and the depletion of hydrocarbons from 100%, 50%, 10% and 1% WAFs at multiple time points over 7 d were measured. Extracts were analysed for THC with GC-FID. Total hydrocarbon content was reported as the sum of hydrocarbons (µg/L) in the range less than n-C9 to C28 (Dataset AAS_3054_THC_WAF). For fertilisation, and 2 cell embryonic development assays that were done in sealed vials, measured values in freshly decanted 50% and 10% WAF dilutions were used as the exposure concentrations. For the embryonic and larval toxicity tests that were done in open vials, the exposure concentrations of THC in WAFs were modelled from the measured concentrations in WAF depletion tests. Exposure concentrations used to model sensitivity estimates were derived by calculating the time weighted mean THC between pairs of successive measurements in the 100% WAFs and dilutions to give overall exposure concentrations for each time point. These modelled concentrations integrated the loss of hydrocarbons over time, and renewal of test solutions at 4 d intervals.

Overview of the project and objectives: To investigate whether nitrate uptake and processes other than nitrate uptake by phytoplankton are significant and show spatial variability possibly induced by varying availability of Fe and other parameters in the region, seawater was collected from CTD (Conductivity, Temperature and Depth) and TMR (Trace Metals Rosette) casts jointly with the nutrient sampling, as well as well as sea-ice collected from Bio ice-core types on Ice Station, for analysis of nitrate d15N, d18O isotopic composition. Results have been interpreted in the light of prevailing nitrate-nutrient concentrations (Belgian team) and N-uptake regimes for the Ice Stations (new vs. regenerated production and nitrification; see Silicon, Carbon and Nitrogen in-situ incubation Metadata file). Methodology and sampling strategy: Samples for isotopic composition of nitrate were collected from the CTD rosette, TMR and Bio ice-core jointly with the nutrient sampling. Sea-ice sampling: sampling strategy follows ice stations deployment via Bio ice-core type. Most of the time we worked close to / directly on the Trace Metal site following precautions concerning TM sampling (clean suits etc.). When we worked close to the TM site, precautions were not such important because we don't need the same drastic precautions for our own sampling. We work together because we want to propose a set of data which helps to characterize the system of functioning in close relation with TM availability (for that, sampling location have to be as close as possible). All samples were filtered on 0.2 microns acrodiscs and kept at -20 degrees C till analysis in the home-based laboratory. We applied the denitrifier method elaborated by Sigman et al. (2001) and Casciotti et al. (2002). This method is based on the isotopic analysis of delta 15N and delta 18O of nitrous oxide (N2O) generated from nitrate by denitrifying bacteria lacking N2O-reductase activity. As a prerequisite the nitrate concentrations need to be known (nutrients analysis in the home lab.) as this sets sample amount provided to the denitrifier community. Briefly, sample nitrate is reduced by a strain of denitrifying bacteria (Pseudomonas aureofaciens) which transform nitrate into N2O, but lack the enzyme to produce N2. N2O is then analysed for N, O isotopic composition by IRMS (Delta V, Thermo) after elimination of CO2, volatile organic carbon and further cryogenic focusing of N2O (Mangion, 2011). Casciotti K.L., D.M.Sigman, M.G. Hastings, J.K. Bohlke and A. Hilkert, 2002. Measurement of the oxygen isotopic composition of nitrate in seawater and freshwater using the denitrifier method, Analytical Chemistry, 74 (19): 4905-4912. Mangion P., 2011. Biogeochemical consequences of sewage discharge on mangrove environments in East Africa, PhD Thesis, Vrije Universiteit Brussel, 208 pp. Sigman D.M., Casciotti K.L., Andreani M., Barford C., Galanter M. and J.K. Bohlke, 2001. A bacterial method for the nitrogen isotopic analysis of nitrate in seawater and freshwater, Analytical Chemistry, 73: 4145-4153.