This metadata record contains the results from bioassays conducted to show the response of the common Antarctic amphipod, Paramoera walkeri to contamination from combinations of Special Antarctic Blend (SAB) diesel, Marine Gas Oil (MGO) and Intermediate Fuel Oil (IFO 180), chemically dispersed with fuel dispersants Ardrox 6120 and Slickgone NS. Fuel only water accommodated fractions (WAF), chemically enhanced water accommodated fractions (CEWAF) and dispersant only treatments were prepared following the methods in Singer et al. (2000) with adaptations from Barron and Ka’aihue (2003). WAF was made using the ratio of 1: 25 (v/v), fuel to filtered seawater (FSW) following the methods of Brown et al. (in prep). Ratios for chemically dispersed treatments were 1: 100 (v/v), fuel to FSW and 1: 20 (v/v) dispersant to fuel. Dispersant only treatments were made using ratios for CEWAF, substituting the fuel component with FSW. Mixes were made in 5 L or 10 L glass aspirator bottles using a magnetic stirrer to achieve a vortex of 20-25% in the FSW before the addition of test media. The same mixing energy was used to prepare all WAFs for enhanced reproducibility and comparability of results (Barron and Ka’aihue, 2003). Mixes were stirred in darkness to prevent bacterial growth for 42 h with an additional settling time of 6 h at 0 plus or minus 1 oC. Extended stirring times were used following the recommendations determined as part of the hydrocarbon chemistry component of this project (Kotzakoulakis, unpublished data). A dilution series of four concentrations were made from the full strength aqueous phase of each mix using serial dilution. WAF test concentrations were 100%, 50%, 20% and 10% while CEWAF concentrations were 10%, 5%, 1% and 0.1%. These concentrations were chosen in order to quantify the mortality curve and allow statistical calculation of LC50 values. To facilitate comparisons of dispersant toxicity in the presence and absence of fuel, dispersant only test concentrations reflected those of CEWAF treatments. WAF was sealed in airtight glass bottles stored at 0 plus or minus 1 oC for a maximum of 3 h before use. Fresh test solutions were prepared every four days to ensure consistent water quality and replace hydrocarbons that adsorbed or evaporated into the atmosphere. Each test concentration was represented by five replicates with five FSW control beakers, with 10 P. walkeri individuals per replicate. Only healthy and active individuals were chosen with a size range of 7.9 plus or minus 0.7 mm for adults and 2.5 plus or minus 0.2 for juveniles measured from the base of the antennae to the widest part of the dorsal curve. Larger individuals and brooding females were not used to avoid unrelated deaths related to age or reproductive state (Sagar, 1980). Beakers were filled to 200 ml and were left open to allow the natural evaporation of lighter monoaromatic hydrocarbon components that would occur during a real spill. A small square of plankton mesh was placed in each jar to provide a substratum to reduce the stress of laboratory conditions and to help to stem cannibalism. Animals were not fed during experiments to avoid hydrocarbons adsorbed onto food pellets being ingested by the amphipods, thereby introducing an additional exposure pathway. Experiments ran for a total of 12 d exposure duration. Experiments were run in cold temperature-controlled cabinets maintained at a temperature of 0 plus or minus 1 oC, fluorescent lights in the cabinets were set to a light regime of 18 h light, 6 h darkness, following the methods in Brown et al. (2017) to reflect Antarctic summer environmental conditions. Lethal and sublethal observations were made at standard ecotoxicology test times of 24 h, 48 h, 96 h, 7 d, 10 d and 12 d, with an additional observation at 8 d coinciding with one of the 4-day water changes. The health status of each individual was classified on a scale of one to four; one showing no effect up to four being mortality. Mortality was determined by a lack of movement and response to stimuli, particularly in the gills. Dead animals were removed and preserved in 80% ethanol at each observation period. Missing amphipods that may have been cannibalised were included in mortality counts as they were likely to have been moribund or already dead when eaten. In order to simulate a repeated pulse pollutant, 90 to 100% of the test solution volume of each beaker was renewed with freshly made test concentrations every four days to replenish hydrocarbons lost through evaporation and adsorption and ensure consistent water quality. Beakers were topped up to 200 ml between water changes with deionised water to maintain water quality parameters. Duplicate 25 ml aliquots of test concentrations were taken at the beginning and end of each experiment in addition to pre and post water change samples. Samples were immediately extracted with 0.7 μm of dichloromethane spiked with an internal standard of BrC20 (1-bromoeicosane) and cyclooctane. Samples were analysed using Gas Chromatography with Flame Ionisation Detection (GC-FID) and mass spectrometry (GC-MS). To determine actual exposure concentrations, four day measured TPH values were used to create a continuous exposure and evaporation profile over the 12 d test period following the methods outlined in Payne et al. (2014) and Brown et al. (2017).

This metadata record contains the results from bioassays conducted to show the response of the Antarctic gastropod, Skenella palludinoides to contamination from combinations of Special Antarctic Blend (SAB) diesel, chemically dispersed with fuel dispersant Ardrox 6120. Fuel only water accommodated fractions (WAF), chemically enhanced water accommodated fractions (CEWAF) and dispersant only treatments were prepared following the methods in Singer et al. (2000) with adaptations from Barron and Ka’aihue (2003). WAF was made using the ratio of 1: 25 (v/v), fuel to filtered seawater (FSW) following the methods of Brown et al. (2017). Ratios for chemically dispersed treatments were 1: 100 (v/v), fuel to FSW and 1: 20 (v/v) dispersant to fuel. Dispersant only treatments were made using ratios for CEWAF, substituting the fuel component with FSW. Mixes were made in 5 L or 10 L glass aspirator bottles using a magnetic stirrer to achieve a vortex of approximately 20% in the FSW before the addition of test media. The same mixing energy was used to prepare all WAFs for enhanced reproducibility and comparability of results (Barron and Ka’aihue, 2003). Mixes were stirred in darkness to prevent bacterial growth for 18 h with an additional settling time of 6 h at 0 plus or minus 1 oC. A dilution series of four concentrations were made from the full strength aqueous phase of each mix using serial dilution. WAF test concentrations were 100%, 50%, 20% and 10% while CEWAF concentrations were 10%, 5%, 1% and 0.1%. These concentrations were chosen in order to quantify the mortality curve and allow statistical calculation of LC50 values. To facilitate comparisons of dispersant toxicity in the presence and absence of fuel, dispersant only test concentrations reflected those of CEWAF treatments. WAF was sealed in airtight glass bottles stored at 0 plus or minus 1 oC for a maximum of 3 h before use. Fresh test solutions were prepared every four days to ensure consistent water quality and replace hydrocarbons that adsorbed or evaporated into the atmosphere. Each test concentration was represented by five replicates with five FSW control beakers, with approximately 10 S.palludinoides individuals per replicate. The healthiest and most active individuals were chosen. Beakers were filled to 200 ml and were left open to allow the natural evaporation of lighter monoaromatic hydrocarbon components that would occur during a real spill. Animals were not fed during experiments to prevent hydrocarbons being ingested, thereby introducing an additional exposure pathway. Experiments ran for a total of 35 d exposure duration for WAF and CEWAF experiments and 15 d for dispersant only experiments. Experiments were run in cold temperature-controlled cabinets set at a temperature of 0 plus or minus 1 oC, fluorescent lights in the cabinets were set to a light regime of 18 h light, 6 h darkness, following the methods in Brown et al. (2017) to reflect Antarctic summer environmental conditions. Lethal and sublethal observations were made at test times of: 24 h, 48 h, 96 h, 7 d, 8 d, 10 d and 12 d, 14 d, 16 d, 20 d, 21 d, 28 d and 35 d for SAB + Ardrox 6120 experiments and 24 h, 48 h, 96 h, 7 d, 8 d, 10 d and 12 d, 14 d, 15 d for Ardrox 6120 only experiments. The health status of each individual was classified as per the criteria listed below: - Attached to the vial with horns in or out - Unattached (often upside down), horns out, will reattach if flipped over - Not attached but if touched, will retract - Closed but attached and out of water - Operculum closed - Dead, operculum open a little (muscles no longer working), if touched, operculum will not move and tissues might disintegrate Dead animals were removed and preserved in 80% ethanol at each observation period. In order to simulate a repeated pulse pollutant, 90 to 100% of the test solution volume of each beaker was renewed with freshly made test concentrations every four days to replenish hydrocarbons lost through evaporation and adsorption and ensure consistent water quality. Beakers were topped up to 200 ml between water changes with deionised water to maintain water quality parameters. Duplicate 25 ml aliquots of test concentrations were taken at the beginning and end of each experiment in addition to pre and post water change samples. Samples were immediately extracted with 0.7 μm of dichloromethane spiked with an internal standard of BrC20 (1-bromoeicosane) and cyclooctane. Samples were analysed using Gas Chromatography with Flame Ionisation Detection (GC-FID) and mass spectrometry (GC-MS). Brown, K.E., King, C.K., Harrison, P.L., 2017. Lethal and behavioural impacts of diesel and fuel oil on the Antarctic amphipod Paramoera walkeri. Environmental Toxicology and Chemistry. Animal collection, 2013 experiments: animals sourced from AAD aquarium, collected in previous seasons. Animal collection, 2014 experiments: January and February 2014 Experiments were conducted at the Marine Research Facility at the Australian Antarctic Division in Kingston, Tasmania. Experiments using SAB fuel and the fuel dispersant Ardrox 6120 were conducted in August and September 2013, with additional experiments conducted in May 2014 using Ardrox 6120 only.

General overview The following datasets are described by this metadata record, and are available for download from the provided URL. - Raw log files, physical parameters raw log files - Raw excel files, respiration/PAM chamber raw excel spreadsheets - Processed and cleaned excel files, respiration chamber biomass data - Raw rapid light curve excel files (this is duplicated from Raw log files), combined dataset pH, temperature, oxygen, salinity, velocity for experiment - Associated R script file for pump cycles of respirations chambers #### Physical parameters raw log files Raw log files 1) DATE= 2) Time= UTC+11 3) PROG=Automated program to control sensors and collect data 4) BAT=Amount of battery remaining 5) STEP=check aquation manual 6) SPIES=check aquation manual 7) PAR=Photoactive radiation 8) Levels=check aquation manual 9) Pumps= program for pumps 10) WQM=check aquation manual #### Respiration/PAM chamber raw excel spreadsheets Abbreviations in headers of datasets Note: Two data sets are provided in different formats. Raw and cleaned (adj). These are the same data with the PAR column moved over to PAR.all for analysis. All headers are the same. The cleaned (adj) dataframe will work with the R syntax below, alternative add code to do cleaning in R. Date: ISO 1986 - Check Time:UTC+11 unless otherwise stated DATETIME: UTC+11 unless otherwise stated ID (of instrument in respiration chambers) ID43=Pulse amplitude fluoresence measurement of control ID44=Pulse amplitude fluoresence measurement of acidified chamber ID=1 Dissolved oxygen ID=2 Dissolved oxygen ID3= PAR ID4= PAR PAR=Photo active radiation umols F0=minimal florescence from PAM Fm=Maximum fluorescence from PAM Yield=(F0 – Fm)/Fm rChl=an estimate of chlorophyll (Note this is uncalibrated and is an estimate only) Temp=Temperature degrees C PAR=Photo active radiation PAR2= Photo active radiation2 DO=Dissolved oxygen %Sat= Saturation of dissolved oxygen Notes=This is the program of the underwater submersible logger with the following abreviations: Notes-1) PAM= Notes-2) PAM=Gain level set (see aquation manual for more detail) Notes-3) Acclimatisation= Program of slowly introducing treatment water into chamber Notes-4) Shutter start up 2 sensors+sample…= Shutter PAMs automatic set up procedure (see aquation manual) Notes-5) Yield step 2=PAM yield measurement and calculation of control Notes-6) Yield step 5= PAM yield measurement and calculation of acidified Notes-7) Abatus respiration DO and PAR step 1= Program to measure dissolved oxygen and PAR (see aquation manual). Steps 1-4 are different stages of this program including pump cycles, DO and PAR measurements. 8) Rapid light curve data Pre LC: A yield measurement prior to the following measurement After 10.0 sec at 0.5% to 8%: Level of each of the 8 steps of the rapid light curve Odessey PAR (only in some deployments): An extra measure of PAR (umols) using an Odessey data logger Dataflow PAR: An extra measure of PAR (umols) using a Dataflow sensor. PAM PAR: This is copied from the PAR or PAR2 column PAR all: This is the complete PAR file and should be used Deployment: Identifying which deployment the data came from #### Respiration chamber biomass data The data is chlorophyll a biomass from cores from the respiration chambers. The headers are: Depth (mm) Treat (Acidified or control) Chl a (pigment and indicator of biomass) Core (5 cores were collected from each chamber, three were analysed for chl a), these are psudoreplicates/subsamples from the chambers and should not be treated as replicates. #### Associated R script file for pump cycles of respirations chambers Associated respiration chamber data to determine the times when respiration chamber pumps delivered treatment water to chambers. Determined from Aquation log files (see associated files). Use the chamber cut times to determine net production rates. Note: Users need to avoid the times when the respiration chambers are delivering water as this will give incorrect results. The headers that get used in the attached/associated R file are start regression and end regression. The remaining headers are not used unless called for in the associated R script. The last columns of these datasets (intercept, ElapsedTimeMincoef) are determined from the linear regressions described below. To determine the rate of change of net production, coefficients of the regression of oxygen consumption in discrete 180 minute data blocks were determined. R squared values for fitted regressions of these coefficients were consistently high (greater than 0.9). We make two assumptions with calculation of net production rates: the first is that heterotrophic community members do not change their metabolism under OA; and the second is that the heterotrophic communities are similar between treatments. #### Combined dataset pH, temperature, oxygen, salinity, velocity for experiment This data is rapid light curve data generated from a Shutter PAM fluorimeter. There are eight steps in each rapid light curve. Note: The software component of the Shutter PAM fluorimeter for sensor 44 appeared to be damaged and would not cycle through the PAR cycles. Therefore the rapid light curves and recovery curves should only be used for the control chambers (sensor ID43). The headers are PAR: Photoactive radiation relETR: F0/Fm x PAR Notes: Stage/step of light curve Treatment: Acidified or control The associated light treatments in each stage. Each actinic light intensity is held for 10 seconds, then a saturating pulse is taken (see PAM methods). After 10.0 sec at 0.5% = 1 umols PAR After 10.0 sec at 0.7% = 1 umols PAR After 10.0 sec at 1.1% = 0.96 umols PAR After 10.0 sec at 1.6% = 4.32 umols PAR After 10.0 sec at 2.4% = 4.32 umols PAR After 10.0 sec at 3.6% = 8.31 umols PAR After 10.0 sec at 5.3% =15.78 umols PAR After 10.0 sec at 8.0% = 25.75 umols PAR This dataset appears to be missing data, note D5 rows potentially not useable information See the word document in the download file for more information.

This data set contains chemical parameters determined for marine sediment samples collected in the 2014-15 summer field season as part of the Thala Valley Long term Monitoring (TV-LTM) project. The aim of this project is to examine changes in the marine benthic ecosystem in the vicinity of Casey station following clean-up of the abandoned Thala Valley waste disposal ('tip') site in 2003-04. The chemical parameters are: (1) 1 M hydrochloric acid-extractable elements (mainly metals) by ICP-AES (inductively coupled plasma - atomic emission spectrometry) (2) water-extractable nutrients by FIA (flow injection analysis) (3) petroleum hydrocarbon fractions (TPH: total petroleum hydrocarbons) and persistent organic pollutants (POPs) - polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) - by GC-FID, GC-ECD and GC-MS (gas chromatography - flame ionization detector / electron capture detector / mass spectrometry), respectively (4) Loss on Ignition at 550 degrees Celsius (LOI; as a proxy for Total Organic Matter) and Dry Matter Fraction (DMF) by gravimetric analysis. Data sets 1, 3 and 4 were obtained for composite samples prepared from the 0-5 cm section of 51 marine sediment cores collected by SCUBA divers from impacted (contaminated) and control (pristine) locations around Casey. Data set 2 was obtained for a subsample of the surface section (0-1 cm) of each of 74 marine sediment cores collected during the same sampling campaign. Sample locations: * Brown Bay (BB) - inner, mid and outer sites * Casey Wharf * McGrady Cove * O'Brien Bay (OB) - OB1, OB2, OB3 sites * Shannon Bay * Wilkes (adjacent to abandoned station) Analytical labs involved: * Wild Lab, AAD, Kingston, Tasmania (data sets 1 and 4; sample preparation for data set 2) * Analytical Services Tasmania (AST), New Town, Tasmania (data set 2) * Analytical Services Unit (ASU), Queen's University, Kingston, Ontario, Canada (data set 3) Information concerning analytical data quality (method reporting limits, accuracy and precision), are included with each data set. Complete analytical method details are available in a separate summary document.