This is a parent metadata record for work carried out as part of ASAC/AAS project 40. See the child metadata records for further information. More than 95% of the biomass in the Southern Ocean is microscopic - single celled plants, animals, bacteria and viruses. We are studying the factors that control their distribution and abundance - oceanographic and seasonal conditions, their physiology, and grazing - in order to model their vital roles as food for other organisms and their influence in moderating global climate change through absorption of CO2 and production of DMS. We are also addressing the changes expected in microbial communities through effects of climate change - global warming, sea ice retreat, ocean acidification and enhanced ultraviolet radiation. This project aims to determine the role of microorganisms in the Southern Ocean. The major objectives are to: * Identify and quantify key protistan components of the Southern Ocean ecosystem and study their autoecology. * Identify environmental and ecological processes that control abundance of key microbial components. * Determine interactions between key microbial components to quantify major pathways of carbon flow. * Determine the activity and viability of bacterioplankton and protists in the Southern Ocean. * Distinguish different microbial communities by identifying key taxa and associations so that processes such as primary production, respiration, grazing and particle flux can be readily parameterised in ecological models. * Determine the effect of elevated CO2 concentrations on microbial populations and processes. Taken from the 2008-2009 Progress Report: Progress against objectives: 1. Ongoing sampling from Astrolabe has continued, with 3 return voyages being sampled for phytoplankton species, chlorophyll a and other pigments, coccolithophorid counts and DNA profiles, in conjunction with measurements of CO2, ocean structure, fluorescence and ocean colour by CSIRO / CRC colleagues. 2. Three sets of minicosm experiments were conducted at Davis station with 7 staff spending 4.5 - 5.5 months on site. These experiments consistently found that acidification caused blooms of nanoplanktonic diatoms and increased bacterial activity, apparently by inhibition of microheterotroph grazers, at the expense of larger cells that are more readily ingested by grazers such as krill. We showed for the first time in Antarctic waters that pCO2 affects the structure and function Antarctic microbial communities in a way that may reduce food availability to large grazers. Over 100 cultures of "winners and losers" from such experiments were isolated and returned to Australia. These will form the basis for further physiological experiments including molecular assays. 3. Submission and acceptance of 8 papers from the BROKE-West cruise (5 as senior author). These showed the interactions between bottom-up (micronutient) top-down (grazing) control in structuring microbial populations in the marginal ice zone. Five biogeographic zones were identified on the basis of species composition, and the productivity was measured for each zone. Microzooplankton grazing experiments found that grazing within that microbial loop consumed a significant proportion of new productivity. In some areas later in the season, all productivity was consumed by microheterotrophs, rather than metazoans such as krill. A time sequence was identified for seeding and development of components of ice edge blooms, subsequent grazing and decline and a mechanism postulated for export of micronutrients (e.g. iron) by grazing and sedimentation that prevents subsequent development of surface water blooms and constrains populations to a deep chlorophyll maximum below the level of a nutricline. 4. Detailed analysis of greater than 30 strains of keystone species Emiliania huxleyi of two morphotypes in conjunction with Clara Hoppe (Masters student, Alfred Wegener Institute) and Suellen Cook (PhD student, University of Tasmania) showed consistent differences between strains in terms of pigmentation, responses to light and genetics. The two morphotypes appear to be adapted to different mixing regimes north and south of the Polar Front; the southern form may represent a new species. For a full list of references associated with this project, see the project link at the provided URL.

From the abstract of some of the papers: It has been suggested that increased springtime UVB radiation caused by stratospheric ozone depletion is likely to reduce primary production and induce changes in the species composition of Antarctic marine phytoplankton. Experiments conducted at Arthur Harbour in the Antarctic Peninsula revealed a reduction in primary productivity at both ambient and increased levels of UVB. Laboratory studies have shown that most species in culture are sensitive to high UVB levels, although the level at which either growth or photosynthesis is inhibited is variable. Stratospheric ozone depletion, with resultant increased springtime UVB irradiance, has been occurring with increasing severity since the late 1970's. Thus the phytoplankton community has already experienced about 20 years' exposure to increasing levels of UVB radiation. Here we present analyses of diatom assemblages from high-resolution stratigraphic sequences from anoxic basins in fjords of the Vestfold HIlls, Antarctica. We find that compositional changes in the diatom component of the phytoplankton community over the past 20 years cannot be distinguished from long-term natural variability, although there is some indication of a decline in the production of some sea-ice diatoms. We anticipate that our results are applicable to other Antarctic coastal regions, where thick ice cover and the timing of the phytoplankton bloom protect the phytoplankton from the effects of increased UVB radiation. Growth rate, survival, and stimulation of the production of UV-B (280 to 320 nm) absorbing compounds were investigated in cultures of five commonly occurring Antarctic marine diatoms exposed to a range of UV-B irradiances. Experimental UV-B exposures ranged from 20 to 650% of the measured peak surface irradiance at an Antarctic coastal site (0.533 J per square metre per second). The five diatom species (Nitzschia lecointei, Proboscia alata, P. inermis, Thalassiosira tumida and Stellarima microtrias) appear capable of surviving two to four times this irradiance. In contrast to Phaeocystis cf. pouchetti, another major component of the Antarctic phytoplankton, the concentrations of pigments with discrete UV absorption peaks in diatoms were low and did not change significantly under increasing UV-B irradiance. Absorbance of UV-B by cells from which pigments had been extracted commonly exceeded that of the pigments themselves. Most of this absorbance was due to oxidisable cell contents, with the frustule providing the remainder. Survival of diatoms did not correlate with absorption by either pigments, frustules or oxidisable cell contents, indicating that their survival under elevated UV-B irradiances results from processes other than screening mechanisms. Springtime UV-B levels have been increasing in Antarctic marine ecosystems since the 1970's. Effects on natural phytoplankton and sea-ice algal communities, however, remain unresolved. At the Marginal Ice Edge Zone, enhanced springtime UV-B levels coincide with a shallow, stratified water column and a major phytoplankton bloom. In these areas it is possible that phytoplankton growth and survival is adversely impacted by enhanced UV-B. In coastal areas, however, the sea ice, which attenuates most of the UV-B before it reaches the water column, remains until December/January, by which time UV-B levels have returned to long-term seasonal averages. Phytoplankton from these areas are unlikely to show long-term changes resulting from the hole in the ozone layer. Fjords of the Vestfold Hills, eastern Antarctica, have anoxic basins which contain high-resolution, unbioturbated sedimentary sequences. Diatom assemblages from these sequences reflect the diatom component of the phytoplankton and sea-ice algal assemblages at the time of deposition. Twenty-year records from these sequences show no consistent record of change in species composition, diversity or species richness. Six-hundred-year records from the same area also show changes in species abundance greater than those seen in the last 20 years. From these records it can be seen that recent changes in diatom abundances generally fall within the limits of natural variability and there is little evidence of recent changes that might be associated with UV-B-induced change.

The actual piece of equipment used was an International Light IL 1700Radiometer equipped with broad band detectors to measure PAR, UV-A and erythemal UV-B. The effects of UV-B radiation on the fatty acid, total lipid and sterol composition and content of three Antarctic marine phytoplankton were examined in a preliminary culture experiment. Exponential growth phase cultures of the diatoms Odontella weissflogii and Chaetoceros simplex and the Haptophyte Phaeocystis antarctica were grown at 2 (plus or minus 1)degrees C and exposed to 16.3 (plus or minus 0.7) W.m-2 photosynthetically active radiation (PAR). UV-irradiated treatments were exposed to constant UV-A (4.39 (plus or minus 0.20) W.m-2) and low (0.37 W.m-2) or high UV-B (1.59 W.m-2). UV-B treatments induced species specific changes in lipid content and composition. The sterol, fatty acid and total lipid content and profiles for O. weissflogii changed little under low UV-B when compared with control conditions (PAR alone), but showed a decrease in the lipid content per cell under high UV-B treatment. In contrast, when P. antarctica was exposed to low UV-B irradiance, storage lipids were reduced and structural lipids increased indicating that low UV-B enhanced cell growth and metabolism. P. antarctica also contained a higher proportion of polyunsaturated fatty acids under low UV-B in comparison with PAR irradiated control cultures. The flagellate life stage of P. antarctica died under high UV-B irradiation. However, exposure of P. antarctica to high UV-B irradiance increased total lipid, triglyceride and free fat. The effect of UV-B irradiances on the lipid content of Antarctic marine phytoplankton is species specific. Changes in ambient UV-B may alter the nutritional quality of food available to higher trophic levels. EXPERIMENTAL All measurements of irradiance were made with an International Light IL 1700 Radiometer equipped with broad band detectors to measure PAR, UV-A and erythemal UV-B [14]. A National Institute of Standards and Technology intercomparison package (NIST Test #534/240436-88) was used to calibrate each light sensor. Unialgal cultures of the diatoms Odontella weissflogii and Chaetoceros simplex were isolated from sea ice collected in Prydz Bay, Antarctica during the 1990/91 austral summer. Phaeocystis antarctica was isolated from Prydz Bay in 1982/83 summer. Cultures of diatoms and Phaeocystis antarctica were maintained in 2 l glass flasks using f/2 growth medium [32] and GP5 medium [33] respectively at a temperature of 2 plus or minus 1 degrees C. Cool white fluorescent lights provided photosynthetically active radiation (PAR) intensity of 17.08 J.m-2.s-1 (84.7 micro E.m-2.s-1), with no UV-B enhancement, on a 12 h light : 12 h dark cycle. Immediately before experimental irradiation, three replicate subsamples of approximately 15 ml were obtained from each parental culture and fixed with Lugols iodine, a known sample volume sedimented, and cells counted over 15 replicate fields using a Labovert inverted microscope. Mean cell concentration and standard deviation were then computed. Each exponential growth phase parental culture was thoroughly mixed and 3 replicate 300 ml Costar polystyrene culture flasks (which completely absorbed wavelengths below 295 nm) established for each light treatment (control, low and high UV exposures). Cultures were irradiated for 24 hours in a 48 hour experimental period (6 h light : 12 h dark : 12 h light : 12 h dark : 6 h light) [14, 23]. Exposures were conducted in a Thermoline controlled environment cabinet at 2 plus or minus 1 degrees C with cool white fluorescent tubes to provide PAR and UV-A (320-400 nm), with UV-B provided by FS20T 12 UV-B Westinghouse sunlamps. PAR and UV-A irradiances were 16.3 plus or minus 0.7 W.m-2 (81.3 plus or minus 3.4 micro E.m-2.s-1) and 4.39 plus or minus 0.20 W.m-2 respectively. The spectral distribution and UV-B irradiance were varied by attenuation with glass filters [5] to provide low (0.37 W.m-2) or high UV-B (1.59 W.m-2). Sensors were each covered by an attenuating glass screen and a single layer of Costar culture flask to measure the experimental irradiances to which the algae were exposed. UV-B irradiances were chosen to reflect less than (74%) and greater than (318%) peak UV-B exposure as measured at an Antarctic coastal site (Casey station, 66 degrees S, [34]). Following irradiation each culture was well mixed and approximately 15 ml was fixed with Lugols Iodine for subsequent estimation of cell concentration (as above). Chlorotic and greatly vesicularised cells were considered to be dead [23]. The remainder of each experimental culture was filtered through Whatman GF/F filters. On completion of filtration, the filters were stored at -20C overnight before extraction of lipids the following day.

A collection of about 20 isolates of Antarctic microalgae from the Windmill Islands region, around Casey Station has been established in the University of Malaya Algae Culture Collection (UMACC). The Antarctic microalgae in the collection includes Chlamydomonas, Chlorella, Stichococcus, Navicula. Ulothrix and Chlorosarcina. Comparative studies on the effect of global warming and UVR stress on these Antarctic microalgae and the tropical collection are being conducted. From the abstract of one of the referenced papers: The growth, biochemical composition and fatty acid profiles of six Antarctic microalgae cultured at different temperatures, ranging from 4, 6, 9, 14, 20 to 30 degrees C, were compared. The algae were isolated from seawater, freshwater, soil and snow samples collected during our recent expeditions to Casey, Antarctica, and are currently deposited in the University of Malaya Algae Culture Collection (UMACC). The algae chosen for the study were Chlamydomonas UMACC 229, Chlorella UMACC 234, Chlorella UMACC 237, Klebsormidium UMACC 227, Navicula UMAC 231 and Stichococcus UMACC 238. All the isolates could grow at temperatures up to 20 degrees C; three isolates, namely Navicula UMACC 231 and the two Chlorella isolates (UMACC 234 and UMACC 237) grew even at 30 degrees C. Both Chlorella UMACC 234 and Stichococcus UMAC 238 had broad optimal temperatures for growth, ranging from 6 to 20 degrees C (growth rate = 0.19 - 0.22 per day) and 4 to 14 degrees C (growth rate = 0.13 - 0.16 per day), respectively. In constrast, optimal growth temperatures for Navicula UMACC 231 and Chlamydomonas UMACC 229 were 4 degrees C (growth rate = 0.34 per day) and 6 to 9 degrees C (growth rate = 0.39 - 0.40 per day), respectively. The protein content of the Antarctic algae was markedly affected by culture temperature. All except Navicula UMACC 231 and Stichococcus UMACC contained higher amount of proteins when grown at low temperatures (6-9 degrees C). The percentage of PUFA, especially 20:5 in Navicula UMACC 231 decreased with increasing culture temperature. However, the percentages of unsaturated fatty acids did not show consistent trend with culture temperature for the other algae studied. There are three spreadsheets available in the download file. ASAC_2590 - provides detail about where each species of algae was collected from. ASAC_2590a - provides data from Teoh Ming-Li et al (2004) ASAC_2590b - provides data from Wong Chiew-Yen et al (2004) The fields in this dataset are: Isolate Culture Collection number Origin (Location) Fatty acids saturated fatty acids polyunsaturated fatty acids monounsaturated fatty acids Temperature growth rate PAR UVB

Environmental manipulation and competition experiments on cultured and natural diatoms will identify the response of key taxa to environment modification. Understanding the environmental factors governing diatom distribution and natural variability will provide a basis to interpret palaeo-environment records, and allow predictions how this temperature-sensitive ecosystem will respond to future change. Environmental manipulation and competition experiments using diatoms will identify the response of key taxa to environment modification. Understanding the environmental factors governing their distribution and natural variability will provide a basis to interpret palaeo-environment records, and allow predictions how this temperature-sensitive ecosystem will respond to future change. Diatoms for the experiments were collected in 2002 (Aurora Australia, Voyage 1) and 2003 (Aurora Australis, Voyage 1). On each occasion water from the ship's online seawater tap was filtered through a 20 micrometre plankton net for up to one hour into a sample jar. A portion of the sample was preserved in lugol's iodine for later phytoplankton analysis, and the rest of the sample maintained alive in the dark in seawater at a constant low temperature. The live sample is maintained at the AAD for culturing and environment manipulation and competition experiments. Project 2364 Twelve water samples were collected from 23/10/03 to 27/10/03, in open seawater between 60 degrees 45' S and 50 degrees 02' S. At each site, the following data were recorded from the ship's data logger: latitude, longitude, UTC time, local time, water depth, salinity, water temperature, fluorescence, UVB, and conductivity.