Data shows gross body measurements of lantern shellfish (Laternula elliptica) collected by divers at McGrady Cove; Brown Bay Inner, and Shannon Bay. Measurements include length, width, and height of shell and weight with shell on and shell off.

Data shows carbon and nitrogen stable isotope concentration in siphon tissue of laternula elliptica from three sites adjacent to Casey Station. McGrady Cove, Brown Bay Inner and Shannon Bay. All shellfish were collected by divers during the 2014/15 summer season. Samples were sent to Cornell University Stable Isotope laboratory for analysis.

Study location and species The four species used in this study were collected from subantarctic Macquarie Island (54.6167 degrees S, 158.8500 degrees E), just north of the Antarctic Convergence in the Southern Ocean. Sea temperatures surrounding Macquarie Island are relatively stable throughout the year, with average temperatures ranging from ~4 to 7 degrees C [25]. Collection sites were free from any obvious signs of contamination and did not have elevated concentrations of metals as confirmed by analysis of seawater samples from the collection sites by inductively coupled plasma optical emission spectrometry (ICP-OES; Varian 720-ES). Toxicity tests were conducted at Macquarie Island over the 2013/14 austral summer, and at the Australian Antarctic Division (AAD) in Tasmania, Australia, from 2013 to 2015. The aquarium at the AAD used for culturing and for holding biota prior to their use in tests was maintained at a temperature of 5.8 degrees C under flow-through conditions (at 0.49L/sec). Individuals for toxicity tests on the island and individuals for return to Australia for culturing were collected from a range of habitats within the intertidal and subtidal zones. All species were highly abundant in each of their respective habitats. The gastropod Laevilittorina caliginosa was collected from pools high on the intertidal zone; the flatworm Obrimoposthia ohlini, from the undersides of boulders from the intertidal to shallow subtidal areas; the bivalve Gaimardia trapesina, from several macroalgae species in high energy locations in the shallow subtidal; and the isopod Limnoria stephenseni, from the floating fronds of the kelp Macrocystis pyrifera, which were located several hundred meters offshore. Test specimens were acclimated to laboratory conditions 24 h to 48 h prior to commencement of tests. Juvenile flatworms, isopods and gastropods were all products of reproduction in the laboratory at the AAD, and hence their approximate age at testing is known. The flatworms hatched from small (2 mm in diameter) brown eggs, laid on rocks or on the side of aquaria. The flatworms exhibited age based morphological differences; juvenile flatworms were light grey in colour, while the adults were black. The gastropods hatched from small (1 mm in diameter) translucent eggs laid on weed, often in a cluster. For flatworms and gastropods, juveniles were not all the same age at testing due to differing hatching times, with ages ranging from 2 weeks to 3 months. In contrast, juvenile isopods were all the same age. Although brooding isopods were not observed, juveniles were noticed during routine feeding, thus were likely within 2-3 days of being released, 6 months after adults were brought from the field to the aquarium. The tests with these juvenile isopods were done within 1 week of their being observed. Care was taken to collect adults from the field, for each species, within a narrow size range to minimise differences in ages between individuals tested (Table 1). However, ages of adults individuals used in tests are unknown. The smaller size class of bivalves tested (juveniles: 2.5 plus or minus 0.5 mm, Table 1) was also collected from the field along with the adults (8.0 plus or minus 1.0 mm, Table 1). Based on knowledge on the growth rate of this species (0.8 mm per year; Everson [26], the smaller size class likely represents a young adult of approximately 2.5 to 4 y old, as opposed to a juvenile stage, and adults collected were approximately 9 to 11 y old. Toxicity tests A static non-renewal test regime was used for all tests. Two replicate tests were done for each species at each life stage, with the exception of the juvenile isopods, where due to the limited number of individuals available, only one test was done. Longer tests durations of 14 days were done for acute responses due to the longer life span and response to contaminants compared to temperate and tropical species as determined in previous studies [7, 27]. All experimental vials and glassware were washed in 10% nitric acid and rinsed thoroughly with MilliQ water three times before use. Tests were done in lidded polyethylene vials of varying sizes, depending on the size and number of individuals in the test (Table 1). Water was not aerated as DO stayed relatively high for tests due to high dissolution rates in cold water. Acid washed and Milli-Q rinsed mesh (600 micron nylon) was provided for isopods to rest on, while no structure was added to vials for the other test species. Test solutions were prepared 24 h prior to the addition of invertebrates. Five copper concentrations in seawater were prepared using a 500 mg/L Univar analytical grade CuSO4 in MilliQ stock solution, plus a control for each test. Seawater was filtered to 0.45 microns, and water quality parameters were measured using a TPS 90-FL multimeter at the start (d 0) and end (d 14) of tests. Dissolved oxygen (DO) was greater than 80% saturation, salinity was 33 to 35 ppt, and pH was 8.1 to 8.3 at the start of tests. Tests were kept in controlled temperature cabinets set at 6 degrees C under 16:8h light:dark during the summer, and 12:12 for tests during the rest of the year (light intensity of 2360 lux). Temperatures within cabinets were monitored throughout the test using Thermochron iButton data loggers. Water samples of each test concentration were taken at the start (day 0) and end of tests (day 14). Samples were filtered through an acid and Milli-Q rinsed, 0.45 microns Minisart syringe filter and acidified with 1% ultra-pure nitric acid before being analysed by ICP-OES to determine dissolved metal concentrations. Measured concentrations at the start of tests were within 96% of nominal target concentrations. Averages between measured concentrations at the start and end of tests were made to estimate exposure concentrations, which were subsequently used in statistical analyses to determine point estimates (Table 2). Both survival and sublethal (behavioural) endpoints were used to determine sensitivity to copper. Vials were checked daily and survival and sublethal responses were observed and recorded on days 1, 2, 4, 7, 10 and 14. Tests were terminated when surviving individuals occurred in less than two concentrations, which was generally at 14 d for all species except for bivalves, in which this occurred sooner (7 to 10 d). Gastropods were scored as dead when their operculum was open and there was no response to stimulus (touch of a probe) on the operculum. Flatworms were scored as dead when there was no movement. Bivalves was scored as dead when there was no movement and when the shells were gaping open due to dysfunctional adductor muscles. Isopods were scored as dead when there was no movement of any appendages. The behavioural endpoint scored for each species was attachment, which indicated healthy and active individuals. For gastropods, this meant the foot was fully extended and attached to experimental vials; for flatworms, the whole body was able to attach (as those affected by copper appeared slightly contracted and could not lie flat); for bivalves, the byssal threads were used to fix individuals to the bottom of the vial, with the siphon also visible and shell slightly open for water exchange; and for isopods, individuals were either holding onto the provided mesh or were swimming, in which case they often reattached to the mesh during observation.

We investigated the toxicity of copper, zinc and cadmium to the following taxa: copepods Tigriopus angulatus (Lang) and Harpacticus sp. (Order Harpacticoida, Family Harpacticidae); flatworm Obrimoposthia ohlini (Bergendal) (Order Seriata, Family Procerodidae); bivalve Gaimardia trapesina (Lamarck) (Order Veneroida, Family Gaimardiidae); sea cucumber Pseudopsolus macquariensis (Dendy) (Order Dendrochirotida, Family Cucumriidae); sea star Anasterias directa (Koeler) (Order Forcipulatida, Family Asteriidae). Sites chosen for the collection of invertebrates for this study were free of obvious signs of metal contamination, as verified by the analysis of seawater samples from collection sites by inductively coupled plasma optical emission spectrometry (ICP-OES). Six invertebrate species were selected for toxicity tests to represent a range of taxa and ecological niches. Individuals of the copepod Tigriopus angulatus were collected using fine mesh dip nets from rock pools high on the intertidal zone. Individuals of the flatworm Obrimoposthia ohlini were collected from the undersides of boulders, high in the intertidal zone. The copepod Harpacticus sp. and bivalve Gaimardia trapesina were collected from several macroalgae species at high energy locations in the intertidal zone. Individuals of the sea cucumber Pseudopsolus macquariensis were collected from rocks from high energy locations from the intertidal to subtidal zones. Juveniles of the sea star Anasterias directa were collected from rocks in deep pools, low in the intertidal zone. All experimental tests using O. ohlini, T. angulatus, P. macquariensis and A. directa were conducted at the AAD Kingston laboratories, while some tests with Harpacticus sp. and all tests with G. trapesina were conducted in the laboratory facilities on Macquarie Island. Adult life-stages were tested for all species except for P. macquairensis and A. directa in which juvenile stages were tested. Psedopsolus macquariensis released eggs in the aquarium which developed into juveniles prior to being used in tests, and juvenile A. directa were collected from the field. Each test involved exposure to copper, zinc or cadmium solution under a static non-renewal test regime over 14 days. Five metal concentrations plus a control were used for each test, with 3-5 replicates of each concentration. Where possible, tests were replicated. Concentrations used in replicate tests sometimes varied, as species sensitivity information accrued in tests was used to optimise subsequent tests. Metal test solutions in seawater were prepared 24 hours prior to the addition of animals, using 500 micrograms/L CuSO4, 500 micrograms/L ZnCl2 and 500 micrograms/L Cd SO4 MilliQ stock solutions. Seawater was filtered to 0.45 microns and water quality parameters were measured using a TPS 90-FL multimeter at the start and end of tests. Dissolved oxygen (DO) was greater than 80% saturation, salinity 35 ppt plus or minus 0.5, and pH was ~8.1-8.3 at the start of tests. All experimental vials and glassware were acid washed with 10% nitric acid and rinsed with MilliQ three times before use. Metal concentrations were determined using ICP-OES; samples of test solutions were taken at the start (day 0) and end of tests (day 14), filtered through a 0.45 microns syringe filter and acidified with 1% ultra-pure nitric acid. Measured concentrations at the start of tests were within 96% of nominal concentrations. In order to estimate exposure concentrations, the measured concentrations at days 0 and 14 were averaged. Tests were conducted in lidded plastic vials of varying sizes, depending on the size and number of individuals in the test. For both copepod species, there were 10 individuals per 50 mL in 70 mL vials; for P. macquariensis there were 8 individuals per 50 mL in 70 mL vials; and for O. ohlini, A. directa and G. trapesina, 10 individuals per 100 mL in 120 mL vials. Tests were conducted under a light-dark regime (at 2360 lux) of 18:6h light:dark in summer, 12:12 for tests for the rest of the year. Tests were kept in controlled temperature cabinets set at 6 degrees C, and temperatures within cabinets were monitored throughout the test using data loggers. Vials were checked daily and survival recorded on days 1, 2, 4, 7, 10 and 14. Individuals were considered dead, and removed from test vials, when for G. trapesina adductor muscles no longer closed shell; O. ohlini were inactive and covered in mucous; P. macquariensis and A. directa tube feet were no longer moving; T. angulatus and Harpacticus sp. urosomes were perpendicular to prosomes. Data are provided in a series of excel workbooks; one workbook per test species.

Study location and test species Subantarctic Macquarie Island lies in the Southern Ocean, just north of the Antarctic Convergence at 54 degrees 30' S, 158 degrees 57' E. Its climate is driven by oceanic processes, resulting in highly stable daily and inter-seasonal air and sea temperatures (Pendlebury and Barnes-Keoghan, 2007). Temperatures in intertidal rock pools (0.5 to 2 m deep) were logged with Thermochron ibuttons over two consecutive summers and averaged 6.5 (plus or minus 0.5) degrees C. The island is relatively pristine and in many areas there has been no past exposure to contamination. To confirm sites used for invertebrate collections were free from metal contamination, seawater samples were taken and analysed by inductively coupled plasma optical emission spectrometry (ICP-OES; Varian 720-ES; S1) The four invertebrate species used in this study were drawn from a range of taxa and ecological niches (Figure 1). The isopod Limnoria stephenseni was collected from floating fronds of the kelp Macrosystis pyrifera, which occurs several hundred meters offshore. The copepod Harpacticus sp. and bivalve Gaimardia trapesina were collected from algal species in the high energy shallow, subtidal zone. Finally, the flatworm Obrimoposthia ohlini was collected from the undersides of boulders throughout the intertidal zone. We hypothesised L. stephenseni would be particularly sensitive to changes in salinity and temperature due to its distribution in the deeper and relatively stable subtidal areas, while O. ohlini would be less sensitive due to its distribution high in the intertidal zone and exposure to naturally variable conditions. We reasoned that the remaining two species, G. trapesina, and Harpacticus sp. were intermediate in the conditions to which they are naturally exposed and hence would likely be intermediate in their response. Test procedure The combined effect of salinity, temperature and copper on biota was determined using a multi-factorial design. A range of copper concentrations were tested with each combination of temperatures and salinities, so that there were up to 9 copper toxicity tests simultaneously conducted per species (Table 1). Experiments on L. stephenseni and Harpacticus sp. were done on Macquarie Island within 2 to 3 days of collection, during which they were acclimated to laboratory conditions. While, G. trapesina and O. ohlini were transported by ship to Australia in a recirculating aquarium system and maintained in a recirculating aquarium at the Australian Antarctic Division in Hobart, both at 6 degreesC. These two taxa were used in experiments within 3 months of their collection. A limited number of G. trapesina and O. ohlini were available, resulting in fewer combinations of stressors tested. Controls for the temperature and salinity treatments were set at ambient levels of 35 plus or minus 0.1 ppt and 5.5 to 6 degreesC for all species. The lowered control temperature for the bivalve reflected the cooler seasonal temperatures at time of testing and lower position within the intertidal. Previous tests conducted under these ambient conditions provided information on the ranges of relevant copper concentrations, appropriate test durations, and water change regimes for each taxon (Holan et al., 2017, Holan et al., 2016b). From these previous studies, we determined that a test duration of 14 d was sometimes required with 7 d often being the best outcome for most species due to high control survival and sufficient response across concentrations. The bivalve G. trapesina was an exception to this due to unfavourable water quality after 3 days in previous work (Holan et al., 2016). For the other three species, this longer duration for acute tests, compared to tests with tropical and temperature species (24 to 96 h) was consistent with previous Antarctic studies that have required longer durations in order to elicit an acute response in biota (King and Riddle, 2001, Marcus Zamora et al., 2015, Sfiligoj et al., 2015). Experimental variables (volume of water, density of test organisms, copper concentrations, temperatures and salinities) differed for each experiment due to differences between each species (Table 1). The temperature increases that were tested (2 to 4 degreesC) reflected the increased sea and air temperatures predicted for the region tested by 2100 (Collins et al., 2013). Treatments were prepared 24 h prior to the addition of animals. Seawater was filtered to 0.45 microns and water quality was measured using a TPS 90-FL multimeter at the start and end of tests. Dissolved oxygen was greater than 80% saturation and pH was 8.1 to 8.3 at the start of tests. All experimental vials and glassware were washed with 10% nitric acid and rinsed with MilliQ water three times before use. Salinity of test solutions was prepared by dilution through the addition of MilliQ water. Copper treatments using the filtered seawater at altered salinities were prepared using 500mg/L CuSO4 (Analytical grade, Univar) in MilliQ water stock solution. Samples of test solutions for metal analysis by ICP-OES were taken at the start and end of tests (on days 0 and 14). Details of ICP-OES procedures are described in the Supplemental material (S4). Samples were taken using a 0.45 µm syringe filter that had been acid and Milli-Q rinsed. Samples were then acidified with 1% diluted ultra-pure nitric acid (65% Merck Suprapur). Measured concentrations at the start of tests were within 96% of nominal concentrations. In order to determine approximate exposure concentrations for each treatment, we averaged the 0 d and 14 d measured concentrations (Table 1). Tests were conducted in temperature controlled cabinets at a light intensity of 2360 lux. At the Macquarie Island station a light-dark regime of 16:8 h was used to mimic summer conditions. In the laboratories in Kingston, Australia, a 12:12 h regime was used to simulate Autum light conditions (as appropriate for the time of testing). Test individuals were slowly acclimated to treatment temperatures over 1 to 2 h before being added to treatments. Temperatures were monitored using Thermochron ibutton data loggers within the cabinets for the duration of the tests. Determination of mortality of individuals differed for each taxon. Mortality was recorded for Gaimardia trapesina when shells were open due to dysfunctional adductor muscles; for Obrimoposthia ohlini when individuals were inactive and covered in mucous; for Limnoria stephenseni when individuals were inactive after gentle stimulation with a stream of water from a pipette; and for Harpacticus sp. when urosomes were perpendicular to prosomes (as used in other studies with copepods; see Kwok and Leung, 2005). All dead individuals were removed from test vials.

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

Human impacts threaten not only species, but also entire ecosystems. Ecosystems under stress can collapse or transition into different states, potentially reducing biodiversity at a variety of scales. Here we examine the vulnerability of shallow invertebrate-dominated ecosystems on polar seabeds, which may be threatened for several reasons. These unique communities consist of dark-adapted animals that rely on almost year-round sea-ice cover to create low-light shallow marine environments. Climate change is likely to cause early sea-ice break-out in some parts of Antarctica, which will dramatically increase the amount of light reaching the seabed. This will potentially result in ecological regime shifts, where invertebrate-dominated communities are replaced by macroalgal beds. Habitat for these endemic invertebrate ecosystems is globally rare, and the fragmented nature of their distribution along Antarctic coast increases their sensitivity to change. At the same time, human activities in Antarctica are concentrated in areas where these habitats occur, compounding potential impacts. While there are clear mechanisms for these threats, lack of knowledge about the current spatial distribution of these ecosystems makes it difficult to predict the extent of ecosystem loss, and the potential for recovery. In this paper we describe shallow ice-covered ecosystems, their association with the environment, and the reasons for their vulnerability. We estimate their spatial distribution around Antarctica using sea-ice and bathymetric data, and apply the IUCN Red List of Ecosystems criteria to formally assess their vulnerability. We conclude that shallow ice-covered ecosystems should be considered near threatened to vulnerable in places, although the magnitude of risk is spatially variable. This dataset comprises two files. Both are provided in netCDF format in polar stereographic project (see nc file for projection details). light_budget_6km.nc : this gives the estimated annual light budget (in mol photons/m^2/year) at the surface of the water column, having been adjusted for sea ice cover (see paper for details). This is calculated on the 6.25km grid associated with the sea ice concentration data. benthic_light_500m.nc : this gives the estimated annual light budget (in mol photons/m^2/year) at the sea floor, having been further adjusted for water depth. It is provided on a 500m grid (as per the IBCSO bathymetry used). Areas deeper than 200m are given no-data values, and areas outside of the coverage of the sea ice grid are assigned a value of -999. See paper for details.

Metadata record for data from AAS (ASAC) project 3134. This metadata record deals specifically with the objectives relating to "Effects of Ocean Warming and Ocean Acidification on the Echinoderm Sterechinus neumayeri". These data are published in the following paper: Ericson, JA and Ho, MA and Miskelly, A and King, CK and Virtue, P and Tilbrook, BD and Byrne, M, Combined effects of two ocean change stressors, warming and acidification, on fertilization and early development of the Antarctic echinoid Sterechinus neumayeri, Polar Biology, online, (online) pp. 1-8. ISSN 0722-4060 (2011) The study examined the effects of decreased seawater pH and increased seawater temperature on Sterechinus neumayeri fertilisation and embryology. Experiments were carried out in the laboratories at Davis Station, Antarctica during the 2010/2011 summer season as part of a wider research programme looking at the effects of climate change and human impacts on Antarctic marine invertebrates. The research was conducted under project 3134 'Vulnerability of Antarctic marine benthos to increased temperatures and ocean acidification associated with climate change'. Public Ocean acidification and warming are global phenomena that will impact marine biota through the 21st century. This project will provide urgently needed predictive information on the likely survivorship of benthic invertebrates in near shore Antarctic environments that is crucial for risk assessment of potential future changes to oceans. As oceans acidify carbonate saturation decreases, reducing the material required to produce marine skeletons. By examining the effects of increased ocean temperature and acidification on planktonic and benthic life stages of both calcifying and non-calcifying ecologically important organisms, predictions can be made on the potential vulnerability of marine biota to climatic change.

Depth related changes in the composition of infaunal invertebrate communities were investigated at two sites in the Windmill Islands around Casey station, East Antarctica, during the 2006/07 summer. Sediment cores (10cm deep x 10cm diameter) were collected from 4 depths (7m, 11m, 17, and 22m) from each of three transects at two sites (McGrady Cove and O'Brien Bay 1). Cores were sieved through a 500 micron mesh and extracted fauna were preserved in 8% formalin and were later counted and identified to species or to morphospecies established through previous infaunal research at Casey. This work was conducted as part of ASAC 2201 (ASAC_2201).

Infaunal marine invertebrates were collected from inside and outside of patches of white bacterial mats from several sites in the Windmill Islands, Antarctica, around Casey station during the 2006-07 summer. Samples were collected from McGrady Cove inner and outer, the tide gauge near the Casey wharf, Stevenson's Cove and Brown Bay inner. Sediment cores of 10cm depth and 5cm diameter were collected by divers using a PVC corer from inside (4 cores) and outside (4 cores) each bacterial patch. The size of each patch varied from site to site. Cores were sieved at 500 microns and the extracted fauna preserved in 4 percent neutral buffered formalin. All fauna were counted and identified to species where possible or assigned to morphospecies based on previous infaunal sampling around Casey. An excel spreadsheet is available for download at the URL given below. The spreadsheet does not represent the complete dataset, and is only the bacterial mat infauna data. Regarding the infauna dataset: - in - in the mat or patch of bacteria and out is in the "normal" sediment surrounding the patch without evidence of any bacterial mat presence. - Patch numbers were allocated to ensure there was no confusion between patches in the same area. - Fauna names are our identification codes for each species. Some we have confirmed identifications for, some not. Species names, where we have them and as we get them, are listed against these codes in the Casey marine soft-sediment fauna identification guide. This work was completed as part of ASAC 2201 (ASAC_2201).