See the referenced paper for additional details. Sampling. Sampling was conducted on board the RSV Aurora Australis during cruise V3 from 20 January to 7 February 2012. This cruise occupied a latitudinal transect from waters north of Cape Poinsett, Antarctica (65_ S) to south of Cape Leeuwin, Australia (37_ S) within a longitudinal range of 113-115_ E. Sampling was performed as described in ref. 29, with sites and depths selected to provide coverage of all major SO water masses. At each surface station, E250-560 l of seawater was pumped from E1.5 to 2.5m depth. At some surface stations, an additional sample was taken from the Deep Chlorophyll Maximum (DCM), as determined by chlorophyll fluorescence measurements taken from a conductivity, temperature and depth probe (CTD) cast at each sampling station. Samples of mesopelagic and deeper waters (E120-240 l) were also collected at some stations using Niskin bottles attached to the CTD. Sampling depths were selected based on temperature, salinity and dissolved oxygen profiles to capture water from the targeted water masses. Profiles were generated on the CTD descent, and samples were collected on the ascent at the selected depths. Deep water masses were identified by the following criteria: CDW 1/4 oxygen minimum (Upper Circumpolar Deep) or salinity maximum (Lower Circumpolar Deep); AABW 1/4 deep potential temperature minimum; AAIW 1/4 salinity minimum 18. The major fronts of the SO, which coincide with strong horizontal gradients in temperature and salinity 19,30, separate regions with similar surface water properties. The AZ lies south of the Polar Front (which was at 51_ S during sampling), whereas the PFZ lies between the Polar Front and the Subantarctic Front. In total, 25 samples from the AZ, PFZ, SAMW, AAIW, CDW and AABW were collected for this study (Fig. 1, Supplementary Data 1). Seawater samples were prefiltered through a 20-mm plankton net, biomass captured on sequential 3.0-, 0.8- and 0.1-mm 293-mm polyethersulphone membrane filters and filters immediately stored at _80 _C31,32. DNA extraction and sequencing. DNA was extracted with a modified version of the phenol-chloroform method 31. Tag pyrosequencing was performed by Research and Testing Laboratory (Lubbock, USA) on a GS FLXb platform (Roche, Branford, USA) using a modification of the standard 926F/1392R primers targeting the V6-V8 hypervariable regions of bacterial and archaeal 16S rRNA genes (926wF: 50-AAA-CTY-AAA-KGA-ATT-GRC-GG-30 , 1,392 R: 50-ACG-GGCGGT-GTG-TRC-30). Denoising, chimera removal and trimming of poor quality read ends were performed by the sequencing facility.

Oceanographic processes in the subantarctic region contribute crucially to the physical and biogeochemical aspects of the global climate system. To explore and quantify these contributions, the Antarctic Cooperative Research Centre (CRC) organised the SAZ Project, a multidisciplinary, multiship investigation carried out south of Australia in the austral summer of 1997-1998. Taken from the abstracts of the referenced papers: In March 1998 we measured iron in the upper water column and conducted iron- and nutrient-enrichment bottle-incubation experiments in the open-ocean Subantarctic region southwest of Tasmania, Australia. In the Subtropical Convergence Zone (~42 degrees S, 142 degrees E), silicic acid concentrations were low (less than 1.5 micro-M) in the upper water column, whereas pronounced vertical gradients in dissolved iron concentration (0.12-0.84 nM) were observed, presumably reflecting the interleaving of Subtropical and Subantarctic waters, and mineral aerosol input. Results of a bottle-incubation experiment performed at this location indicate that phytoplankton growth rates were limited by iron deficiency within the iron-poor layer of the euphotic zone. In the Subantarctic water mass (-46.8 degrees S, 142 degrees E), low concentrations of dissolved iron (0.05-0.11 nM) and silicic acid (less than 1 micro-M) were measured throughout the upper water column, and our experimental results indicate that algal growth was limited by iron deficiency. These observations suggest that availability of dissolved iron is a primary factor limiting phytoplankton growth over much of the Subantarctic Southern Ocean in the late summer and autumn. The importance of resource limitation in controlling bacterial growth in the high-nutrient, low-chlorophyll (HNLC) region of the Southern Ocean was experimentally determined during February and March 1998. Organic- and inorganic-nutrient enrichment experiments were performed between 42 degrees S and 55 degrees S along 141 degrees E. Bacterial abundance, mean cell volume, and [3H]thymidine and [3H]leucine incorporation were measured during 4- to 5-day incubations. Bacterial biomass, production, and rates of growth all responded to organic enrichments in three of the four experiments. These results indicate that bacterial growth was constrained primarily by the availability of dissolved organic matter. Bacterial growth in the subtropical front, subantarctic zone, and subantarctic front responded most favourably to additions of dissolved free amino acids or glucose plus ammonium. Bacterial growth in these regions may be limited by input of both organic matter and reduced nitrogen. Unlike similar experimental results in other HNLC regions (subarctic and equatorial Pacific), growth stimulation of bacteria in the Southern Ocean resulted in significant biomass accumulation, apparently by stimulating bacterial growth in excess of removal processes. Bacterial growth was relatively unchanged by additions of iron alone; however, additions of glucose plus iron resulted in substantial increases in rates of bacterial growth and biomass accumulation. These results imply that bacterial growth efficiency and nitrogen utilisation may be partly constrained by iron availability in the HNLC Southern Ocean. The download file also contains three excel spreadsheets of iron data from the project. The file Sedwick_A9706_Fe_data contains water-column dissolved Fe and total-dissolvable Fe data from cruise A9706, which is presented in Sedwick et al. (1999) and Sedwick et al. (2008). The files Sedwick_A9706_ProcessStn1_Exp_data and Sedwick_A9706_ProcessStn2_Exp_data present data from shipboard experiments conducted during cruise A9706 at Process Stations 1 and 2, respectively, as reported in Sedwick et al. (1999).

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

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 dataset is a collection of marine environmental data layers suitable for use in Southern Ocean species distribution modelling. All environmental layers have been generated at a spatial resolution of 0.1 degrees, covering the Southern Ocean extent (80 degrees S - 45 degrees S, -180 - 180 degrees). The layers include information relating to bathymetry, sea ice, ocean currents, primary production, particulate organic carbon, and other oceanographic data. An example of reading and using these data layers in R can be found at https://australianantarcticdivision.github.io/blueant/articles/SO_SDM_data.html. The following layers are provided: Layer name: depth Description: Bathymetry. Downloaded from GEBCO 2014 (0.0083 degrees = 30sec arcmin resolution) and set at resolution 0.1 degrees. Then completed with the bathymetry layer manually corrected and provided in Fabri-Ruiz et al. (2017) Value range: -8038.722 - 0 Units: m Source: This study. Derived from GEBCO URL: https://www.gebco.net/data_and_products/gridded_bathymetry_data/ Citation: Fabri-Ruiz S, Saucede T, Danis B and David B (2017). Southern Ocean Echinoids database_An updated version of Antarctic, Sub-Antarctic and cold temperate echinoid database. ZooKeys, (697), 1. Layer name: geomorphology Description: Last update on biodiversity.aq portal. Derived from O'Brien et al. (2009) seafloor geomorphic feature dataset. Mapping based on GEBCO contours, ETOPO2, seismic lines). 27 categories Value range: 27 categories Units: categorical Source: This study. Derived from Australian Antarctic Data Centre URL: https://data.aad.gov.au/metadata/records/Polar_Environmental_Data Citation: O'Brien, P.E., Post, A.L., and Romeyn, R. (2009) Antarctic-wide geomorphology as an aid to habitat mapping and locating vulnerable marine ecosystems. CCAMLR VME Workshop 2009. Document WS-VME-09/10 Layer name: sediments Description: Sediment features Value range: 14 categories Units: categorical Source: Griffiths 2014 (unpublished) URL: http://share.biodiversity.aq/GIS/antarctic/ Layer name: slope Description: Seafloor slope derived from bathymetry with the terrain function of raster R package. Computation according to Horn (1981), ie option neighbor=8. The computation was done on the GEBCO bathymetry layer (0.0083 degrees resolution) and the resolution was then changed to 0.1 degrees. Unit set at degrees. Value range: 0.000252378 - 16.94809 Units: degrees Source: This study. Derived from GEBCO URL: https://www.gebco.net/data_and_products/gridded_bathymetry_data/ Citation: Horn, B.K.P., 1981. Hill shading and the reflectance map. Proceedings of the IEEE 69:14-47 Layer name: roughness Description: Seafloor roughness derived from bathymetry with the terrain function of raster R package. Roughness is the difference between the maximum and the minimum value of a cell and its 8 surrounding cells. The computation was done on the GEBCO bathymetry layer (0.0083 degrees resolution) and the resolution was then changed to 0.1 degrees. Value range: 0 - 5171.278 Units: unitless Source: This study. Derived from GEBCO URL: https://www.gebco.net/data_and_products/gridded_bathymetry_data/ Layer name: mixed layer depth Description: Summer mixed layer depth climatology from ARGOS data. Regridded from 2-degree grid using nearest neighbour interpolation Value range: 13.79615 - 461.5424 Units: m Source: https://data.aad.gov.au/metadata/records/Polar_Environmental_Data Layer name: seasurface_current_speed Description: Current speed near the surface (2.5m depth), derived from the CAISOM model (Galton-Fenzi et al. 2012, based on ROMS model) Value range: 1.50E-04 - 1.7 Units: m/s Source: This study. Derived from Australian Antarctic Data Centre URL: https://data.aad.gov.au/metadata/records/Polar_Environmental_Data Citation: see Galton-Fenzi BK, Hunter JR, Coleman R, Marsland SJ, Warner RC (2012) Modeling the basal melting and marine ice accretion of the Amery Ice Shelf. Journal of Geophysical Research: Oceans, 117, C09031.

Krill-associated bacterial communities characterised by high-throughput DNA sequencing of the 16S ribosomal RNA gene. The data is decribed in 'Clarke LJ, Suter L, King R, Bissett A and Deagle BE (2019) Antarctic Krill Are Reservoirs for Distinct Southern Ocean Microbial Communities. Front. Microbiol. 9:3226. doi: 10.3389/fmicb.2018.03226' available here: https://www.frontiersin.org/articles/10.3389/fmicb.2018.03226/full

Large volumes of water (200 - 500 L) were filtered and fractionated by size for various planktonic components: eukaryotic phytoplankton, prokaryotic picoplankton, and marine viruses. Sample sites were chosen to generate the widest diversity, and included planktonic blooms, oligotrophic zones, small polynyas near sea ice, nearshore areas, and Antarctic bottom water from coastal, canyon and deepwater areas. Half of each sample will be used for DNA library construction, and the other half will be used for meta-proteomic analysis. Random shotgun sequencing of the marine genomic libraries should produce a metagenomic snapshot of planktonic life in a variety of marine habitats. This work was completed as part of ASAC project 2899 (ASAC_2899).

Marine sediment meiofauna community composition and sediment environmental data collected in 2005 and published in Stark, J. S., M. Mohammad, A. McMinn, and J. Ingels. 2020. Diversity, abundance, spatial variation and human impacts in marine meiobenthic nematode and copepod communities at Casey station, East Antarctica. Frontiers in Marine Science 7:480. From the abstract: The composition, spatial structure, diversity and abundance of Antarctic nematode and copepod meiobenthic communities was examined in shallow (5 – 25 m) marine coastal sediments at Casey Station, East Antarctica. The sampling design incorporated spatial scales ranging from 10 meters to kilometres and included testing for human impacts by comparing disturbed (metal and hydrocarbon contaminated sediments adjacent to old waste disposal sites) and control areas. A total of 38 nematode genera and 20 copepod families were recorded with nematodes being dominant, comprising up to 95% of the total abundance. Variation was greatest at the largest scale (km’s) but each location had distinct assemblages. At smaller scales there were different patterns of variation for nematodes and copepods. There were significant differences between communities at control and disturbed locations. Community patterns had strong correlations with concentrations of anthropogenic metals in sediments as well as sediment grain size and total organic content. Given the strong association with environmental patterns, particularly anthropogenic disturbance, meiofauna may be seen as very useful indicators of natural and anthropogenic environmental changes in Antarctica. Methods derived from: Stark, J. S., M. Mohammad, A. McMinn, and J. Ingels. 2020. Diversity, abundance, spatial variation and human impacts in marine meiobenthic nematode and copepod communities at Casey station, East Antarctica. Frontiers in Marine Science 7:480. Sampling design Sampling was undertaken using a hierarchical, nested design with three spatial scales, Locations (separated by kms); within each location there were two sites (~ 100 m apart) and at each site there were two plots (~10m apart). Within each plot (1m diameter), two replicate cores were taken for meiofauna and two for environmental analysis, making a total of 8 meiofauna and 8 environmental cores per location, except at O’Brien Bay-5 where one meiofauna core was lost during sampling. Six locations were sampled around Casey Station. There were three control locations, two of which were within O’Brien Bay to the south of Casey (O’Brien Bay-1 (OB-1) and O’Brien Bay-5 (OB-5)); and one within Newcomb Bay, in McGrady Cove (Fig. 1). There were three locations adjacent to waste disposal sites: two locations were situated along a gradient of pollution within Brown Bay (Inner and Middle)(Stark et al. 2004, Stark 2008); and a third location was at Wilkes, adjacent to the abandoned waste disposal site at the derelict Wilkes station (Stark et al. 2003a), all within Newcomb Bay (Fig. 1). These waste disposal sites were used historically to dispose of all waste and rubbish generated on station and included used oil, building materials, electronics and batteries, food, clothing and chemicals (Snape et al. 2001, Stark et al. 2006). Both waste disposal sites are contaminated with metals and hydrocarbons above background levels (Stark et al. 2008, Stark et al. 2014b, Fryirs et al. 2015). Sample collection, meiofauna preparation and identification Sediment samples were collected by divers using modified 60 ml syringes with their intake end cut off to form a small core tube (28mm internal diameter). Cores were pushed into the sediment to a depth of 10 cm, extracted, and the bottom end was capped. In a few cases samples were only taken down to 5-7 cm, where sediments were less than 10 cm deep due to underlying rock. No sediments less than 5 cm deep were sampled. Cores were transported to Casey Station laboratories where they were emptied into sample jars and 4% formalin was added

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