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See the download file for the full, unedited summary - this version has been slightly modified in order to display correctly. Sample collection and test preparation This metadata record contains the results of four experiments that were conducted to determine the sensitivity and response of Antarctic moss and terrestrial algae to Special Antarctic Blend (SAB) fuel contamination. The work was conducted under AAS Project 4100, with plant material collected from the Casey region during the 2012/2013 field season and subsequent laboratory tests and analyses conducted at Casey station, University of Wollongong and AAD Kingston laboratories, with laboratory work completed in 2013. Four test species were selected for this study, they are all known moss species for the Windmill Islands region and a terrestrial algae, they are all commonly found in ice-free refuges in the vicinity of Casey station. The species were the three mosses: Schistidium antarctici, Ceratodon purpureus, and Bryum pseudotriquetrum, and one terrestrial green alga, Prasiola crispa. Samples of Australian C. purpureus (collected from Wollongong at 34 degrees 24'14"S, 150 degrees 52'30"E) and referred to as 'local C. purpureus') were also used in toxicity tests since this cosmopolitan species may provide valuable insights into potential differences in response to fuel contamination for the same moss species growing in temperate versus polar regions. Four experiments were conducted in this project, one at Casey station 'Optimisation of test parameters and growth conditions' and three at the University of Wollongong, 'Fuel toxicity trial', 'Definitive toxicity test 1 - aged SAB' and 'Definitive toxicity test 2 - fresh SAB'. Experiments were conducted using two different artificial laboratory formulated soil types: a standard soil mixture developed by Environment Canada specifically for soil toxicity testing with plants, and a sandy soil mixture which was developed to more closely represent the properties of soil at Casey station. The two soil mixtures are referred to as 'Canadian standard' and 'Sandy'. Both types were included in the first two experiments, and sandy only was used in the latter two. Canadian standard soil was prepared according to standard methods (Environment Canada 2005), with some small modifications: 10% (w/w) Sphagnum sp. peat, which was autoclaved and oven dried at 70 degrees C overnight, ground and sieved through a 2 mm mesh screen. This was combined with 20% (w/w) kaolin clay (particles less than 40 microns), and 70% (w/w) propagating sand (sieved to 1 mm particles). Sandy soil was prepared using 20% (w/w) kaolin clay and 80% (w/w) sand. The ingredients for both soils, in their dry form, were mixed thoroughly by hand to create homogenous mixes. In the fuel toxicity tests (latter three experiments) soils were spiked with SAB fuel and tumbled overnight in a mechanical sample rotator (Environmental Express, 12 places LE rotator) to allow for thorough mixing. To standardise spiking methods, the amount of SAB fuel required was determined on a soil dry weight basis. A stock of spiked soils were stored in 2.5 L amber schott bottles in a modified fridge (Orford FMF30) at 15 degrees C (plus or minus 1 degrees C) and aged for two weeks for use in tests requiring SAB contaminants that had been aged (fuel toxicity trial and first definitive toxicity test). For all experiments, except the first definitive toxicity test, two different water level treatments were utilised. These water level treatments consisted of: approximately 70% moisture content for the Canadian standard and 17% moisture content for the Casey sandy soils ('low water'); or fully saturated soils with water almost up to the tip of the moss shoots ('high water'). For all experiments, treatments were set up in a similar manner. For each replicate, approximately 50 mL of artificial soil was spread evenly and flattened within a glass petri dish (90 x 15 mm). One small sample (approximately 0.5 cm2) of each species was placed in an indentation in the soil inside a 10 mm stainless steel washer. Petri dishes were placed within sealable transparent plastic containers (Sistema, 5 L, 21.0 x 24.2 x 10.5 cm). All petri dishes were kept in a temperature controlled cabinet (Thermoline Scientific) at a constant temperature of 15 degrees C (plus or minus 1 degrees C) and a day/night photoperiod of 16/8 hours. Light was provided by 16 standard fluorescent tubes (Polylux XLR F18W/840) producing a maximum light intensity of 50 to 55 micro mols/m2/s inside the plastic containers. The temperature and photoperiod were chosen to align with moss turf conditions during summer months in Antarctica and to stimulate moss and algal photosynthesis and growth in order to detect potential effects of exposure to petroleum hydrocarbons. Toxicity tests The first experiment was conducted at Casey Station in January 2013 and tested the growth conditions to be used in later toxicity testing. The four Antarctic species were used in this experiment. The experiment had two treatments: soil type (Canadian standard or Casey sandy) and water level (low or high), and was conducted over 21 days. Chlorophyll fluorescence (Fv/Fm and Fo) measurements were taken to assess plant health. Measurements were taken on the first day of the experiment to determine initial plant health and subsequently after 1, 3, 7, 11 and 21 days exposure to test conditions. All other experiments were conducted at the School of Biological Sciences at the University of Wollongong. The second experiment was conducted to identify suitable methods for testing the sensitivity of Antarctic moss and terrestrial algae to fuel contamination and to identify conditions which provoke a response to SAB fuel. All five plant types (S. antarctici, C. purpureus, B. pseudotriquetrum, P. crispa and local C. purpureus) were used in this toxicity trial. During the first seven days there were two treatments: fuel concentration (ten nominal treatment concentrations of SAB in soils: 40,000, 35,000, 30,000, 25,000, 20,000, 10,000, 5,000, 2,500, 1,250 mg SAB/kg soil and control (0 mg SAB/kg soil)) and soil type (two treatments: Canadian standard and Sandy), with four replicates per treatment. After one week, there was no response to the fuel, therefore another treatment, water level, was added. Half of the replicates received high water levels and the other half continued with low water levels. This resulted in two replicates per treatment. Following another 7 days there was still no response to fuel, therefore a fourth treatment was added, humidity level. The petri dish lids were removed from half of the replicates creating a drier environment. This resulted in no replication amongst treatments. The experiment was conducted for another 7 days, resulting in a total of 21 days exposure to SAB. The third experiment, 'Definitive toxicity test 1 - aged SAB', used pre-desiccated plant samples (to mimic Antarctic field conditions, where there is predominantly a lack of free water, causing moss and terrestrial algae to periodically desiccate). High water levels were used in the test to enhance water (and potentially SAB fuel) uptake. Sandy soil was used as the substrate, as the previous method development tests demonstrated no difference between soil types and the sandy soil better represents the soil at Casey station, and is therefore more relevant for site-specific assessments. The experiment had one treatment, SAB fuel, with six nominal concentration treatments (60,000, 50,000, 40,000, 30,000, 20,000 and 0 (control) mg SAB/kg soil). Actual SAB fuel concentrations were analysed using gas chromatography (GC) as described below and reported as mg total petroleum hydrocarbons (TPH)/kg soil. Measured fuel concentrations for these nominal treatments were 62,900, 48,800, 35,800, 25,500, 17,200 and 0 mg TPH/kg. All five plant types were tested, the experiment had six replicates and was conducted for 21 days. Fv/Fm and Fo measurements were measured daily for the first week then every third day until termination of the experiment at 21 days. At the completion of the experiment moss leaf tip and algal material were collected from each replicate for photosynthetic pigment extraction. The fourth experiment, 'Definitive toxicity test 2 - fresh SAB', was conducted at the University of Wollongong. This experiment was set up as for the first definitive toxicity test with the following four modifications: (1) freshly spiked soils were used, (2) seven concentrations were tested (nominal concentrations of 60,000, 50,000, 40,000, 30,000, 20,000, 10,000 and 0 (control) mg SAB/kg soil and actual fuel concentration treatments were 61,800, 51,900, 40,400, 27,900, 16,300, 6,700 and 0 mg TPH/kg), (3) two water levels (high water and low water), (4) exposure period of 28 days. All five species were tested, the experiment had six replicates and was conducted for 21 days. Fv/Fm and Fo measurements were taken daily for the first week then every third day until termination of the experiment at 28 days. At the completion of the experiment moss leaf tip and algal material were collected from each replicate for photosynthetic pigment extraction. Laboratory Analyses Plant material collected for determination of photosynthetic pigment contents were analysed at the University of Wollongong. Soil samples were analysed for total petroleum hydrocarbons (TPH) at AAD Kingston Laboratories. Photosynthetic pigment extraction: Samples were freeze dried overnight (in a CHRIST Alpha 1-2 LD plus Freeze Drier) and then weighed. Samples were ground for 2 min, with two 2 mm tungsten carbide beads, at 30 Hz in a tissue lyser (Qiagen Retsch po138387). A 0.6 mL aliquot of 60% ethylacetate:40% acetone was added to each sample and samples were then ground for another 2 min at 30 Hz after which 0.5 mL of MilliQ water was added to each sample. Samples were centrifuged for 5 min at 3,600 g (Eppendorf 5415D centrifuge) to separate the pigment extract and aqueous layers. The supernatant was transferred to a 1.5 mL tube, residual liquid was discarded and the carbide beads were removed. A 0.05 mL aliquot of supernatant was made up to 1 mL with 80% acetone:20% MilliQ water. Absorbance at wavelengths of 750, 663.2, 646.8 and 470 nm was measured using a UV-Visible spectrophometer (Shimadzu UV-1601). Total chlorophyll and carotenoid levels as well as carotenoid/chlorophyll ratios in extracts were then determined using equations from Lichtenthaler H. K. and Buschmann C. (2001) Chlorophylls and carotenoids: measurement and charcterization by UV-vis spectroscopy. Current Protocols in food analytical chemistry, F4.3.1-F4.3.8, which can be found on the web (http://www.thyssen-web.de/assets/files/fd_documents/sp_buche/uv_vis_pigmente.pdf). Total petroleum hydrocarbon analysis Spiked soils were sampled for analysis of TPH concentrations by GC. Approximately 12 g of soil was sampled from each concentration and added to a 40 mL headspace vial. To each vial, 10 mL of hexane and 10 mL of MilliQ water was added. Following this, 1 mL of internal standard mix (containing 50 micro grams/mL 1,4-dichlorobenzene, 50 micro grams/mL p-terphenyl, 250 micro grams/mL cyclooctane, 50 micro grams/mL C24D50 and 250 micro grams/mL bromoeicosane) was added to each vial. Samples were shaken vigorously to ensure thorough mixing and tumbled end over end for approximately 17 hours at room temperature. Once extracted, the samples were centrifuged at 1,000 rpm for 5 min to achieve complete separation of the hexane and aqueous layers. A 1.5 mL aliquot of the hexane layer was removed using a glass pipette and transferred to a GC vial for analysis. The remainder of the hexane was transferred to an 8 mL glass vial with a Teflon septa for frozen storage until completion of analysis. Extracts were analysed for TPH on an Agilent 6890N GC-Flame Ionisation Detector (GC-FID) fitted with a split/splitless injector and an auto-sampler (Agilent 7683 ALS). Separation was performed on a SGE BP1 column (25 m x 0.22 mm Internal Diameter, 0.25 microns film thickness). Extracts (3 micro litres) were injected with a 15:1 pulsed split at 310 degrees C and 18.71 psi (pulse pressure 30.0 psi). Helium was used as the carrier gas. Carrier gas velocity at the injector was 23.9 mL/min. At the column, gas flow was held at 1.3 mL/min for 17 min, and then increased to 3.0 mL/min for the duration of the oven program. The GC oven temperature program was 50 degrees C for 3 min, then an increase to 320 degrees C at 18 degrees C/min. Detector temperature was 330 degrees C. Total petroleum hydrocarbon concentrations reported were the SAB fuel hydrocarbon range of n-C9 to n-C18. Description of data files provided The excel spreadsheet 'Standard growth condition test and Fuel toxicity trial.xls' contains the dataset from these first two experiments. The worksheet labelled 'Test conditions Std growth cond' show details of test name, dates, species collection, soil types and test conditions for the experiment 'Optimisation of test parameters and growth conditions'. The worksheet 'Raw data Std growth cond' contains Fv/Fm data. The worksheet consists of one column each for: species, soil type, water level and replicate number, followed by six columns with Fv/Fm measurements collected on day 0, 1, 3, 7, 11 and 21. The worksheet 'Means StD StE Std growth cond' has calculations of means, standard deviation and standard error. Also included are graphs showing means and standard error for the exposure period. The worksheet labelled 'Test conditions Toxicity trial' shows details of test name, dates, species collection, soil types, spiked soil preparation, and test conditions for the experiment 'Fuel toxicity trial'. The worksheet 'Raw data Toxicity trial' contains Fv/Fm and Fo data. The worksheet consists of one column each for the treatment conditions: nominal SAB fuel concentration, species, soil type, replicate, water level and humidity level, followed by data columns providing Fv/Fm and Fo measurements collected daily for 21 days. The worksheet 'Means StD StE Stand cond Tox tr' contains calculations for means, standard deviation and standard error for Fv/Fm for the first week of the experiment when there were two treatments (SAB concentration and soil type) and four replicates. This worksheet also contains graphs showing means and standard error for Fv/Fm on day 7. The worksheet 'Means StD StE Water Lev Tox Tri' contains calculations of means, standard deviation and standard error for Fv/Fm for the second week of the experiment when there were three treatments (SAB concentration, soil type and water level) and two replicates. This worksheet also contains graphs showing means and standard error for Fv/Fm on day 14. The excel spreadsheet 'Definitive toxicity test 1 - aged SAB' contains data from the third experiment, Definitive toxicity test 1 - aged SAB. The worksheet labelled 'Test conditions' shows details of test name, dates, species collection, soil types and test conditions for the experiment. The worksheet labelled 'Raw data chlorophyll fluorescence' contains columns with all Fv/Fm and Fo data collected throughout the experiment. This worksheet also has columns for nominal fuel concentration, actual fuel concentration, species and replicate number. The worksheet 'Means StDev StErr FvFm' has calculations of means, standard deviation and standard error for Fv/Fm measurements taken throughout the experiment. The worksheet 'Graphs Fv/Fm' contains graphs showing means and standard error for Fv/Fm on day 21. The worksheet 'Raw data and calculations pigm' contains chlorophyll a and b and carotenoid data. The column headings are: - nominal fuel concentration (mg SAB/kg soil) - the spiked fuel concentration - actual fuel concentration (mg TPH/kg soil)- the actual measured fuel concentrations determined from GC - replicate number - species - tube (g) - weight of empty Eppendorf tubes, in grams, in which samples were collected for pigment extraction - tube + dry (g) - weight of tube + dry moss/algal sample in grams - dry sample (g) - weight of dry sample in grams - dry sample (mg) - weight of dry sample in milligrams - dilution - dilution level of extracted sample to 80%acetone:20% MilliQ water used for spectrometry measurements - spectrometry readings at 750, 663,2, 646,8 and 470nm - columns for calculations to determine chlorophyll a, chlorophyll b, total chlorophyll (a+b), chlorophyll a:b ratio, total carotenoids and carotenoid:chlorophyll ratio. - Calculations were based on equations from Lichtenthaler H. K. and Buschmann C. (2001). The worksheet 'Means StDev StErr Pigment' has calculations of means, standard deviation and standard error for chlorophyll a, chlorophyll b, total chlorophyll (a+b), chlorophyll a:b ratio, total carotenoids and carotenoid:chlorophyll ratio for day 21. The column headings are: - nominal fuel concentration (mg SAB/kg soil) - the spiked fuel concentration - actual fuel concentration (mg TPH/kg soil)- the actual measured fuel concentrations determined from GC - replicate number - species - Chlorophyll a - amount of chlorophyll a in sample - Chlorophyll b - amount of chlorophyll b in sample - chl a (micro mol.g/dw) - chlorophyll a expressed as a concentration - chl b (micro mol.g/dw) - chlorophyll b expressed as a concentration - Total carotenoids - amount of carotenoids in sample - Total carotenoids (micro mol.g/dw) - carotenoids expressed as a concentration - Chlorophyll a:b - chlorophyll a to chlorophyll b ratio - Total chlorophyll (micro mol.g/dw) - total chlorophyll (a+b) expressed as a concentration - Carotenoid:Chlorophyll - carotenoids to total chlorophyll (a+b) ratio The worksheet 'Graphs Total Chlorophyll' contains graphs showing means and standard error for Total chlorophyll (a+b) (micro mol.g/dw) on day 21. The worksheet 'Graphs Total Carotenoids' contains graphs showing means and standard error for total carotenoids (micro mol.g/dw) on day 21. The worksheet 'Car Chl Ratio' contains graphs showing means and standard error for the carotenoid to chlorophyll ratio on day 21. The excel spreadsheet 'Definitive toxicity test 2 - fresh SAB' contains data from the fourth experiment, Definitive toxicity test 2 - fresh SAB. The worksheet labelled 'Test conditions' show details of test name, dates, species collection, soil types and test conditions for the experiment. The worksheet labelled 'Raw data chlorophyll fluorescence' contains columns with all Fv/Fm and Fo data collected throughout the experiment. This worksheet also has columns for nominal fuel concentration, actual fuel concentration, species, water level and replicate number. The worksheet labelled 'Means StDev StErr FvFm' has calculations of means, standard deviation and standard error for Fv/Fm measurements taken throughout the experiment. The worksheet labelled 'Graphs Fv/Fm' contains graphs showing means and standard error for Fv/Fm on day 28. The worksheet 'Raw data and calculations pigm' contains chlorophyll a and b and carotenoid data. The column headings are: - nominal fuel concentration (mg SAB/kg soil) - the spiked fuel concentration - actual fuel concentration (mg TPH/kg soil)- the actual measured fuel concentrations determined from GC - replicate number - species - water level - high water or low water treatment - tube (g) - weight of empty Eppendorf tubes, in grams, in which samples were collected for pigment extraction - tube + wet (g) - weight of tube + wet moss/algal sample in grams - wet sample (g) - weight of wet sample in grams - tube + dry (g) - weight of tube + dry moss/algal sample in grams - dry sample (g) - weight of dry sample in grams - dry sample (mg) - weight of dry sample in milligrams - dilution - dilution level of extracted sample to 80%acetone:20% MilliQ water used for spectrometry measurements - spectrometry readings at 750, 663.2, 646.8 and 470nm - columns for calculations to determine chlorophyll a, chlorophyll b, total chlorophyll (a+b), chlorophyll a:b ratio, total carotenoids and carotenoid:chlorophyll ratio. - Calculations were based on equations from Lichtenthaler H. K. and Buschmann C. (2001). The worksheet 'Means StDev StErr Pigment' has calculations of means, standard deviation and standard error for chlorophyll a, chlorophyll b, total chlorophyll (a+b), chlorophyll a:b ratio, total carotenoids and carotenoid:chlorophyll ratio for day 21. The column headings are: - nominal fuel concentration (mg SAB/kg soil) - the spiked fuel concentration - actual fuel concentration (mg TPH/kg soil)- the actual measured fuel concentrations determined from GC - replicate number - species - water level - high water or low water - Chlorophyll a - amount of chlorophyll a in sample - Chlorophyll b - amount of chlorophyll b in sample - chl a (micro mol.g/dw) - chlorophyll a expressed as a concentration - chl b (micro mol.g/dw) - chlorophyll b expressed as a concentration - Chlorophyll a:b - chlorophyll a to chlorophyll b ratio - Total chlorophyll (micro mol.g/dw) - total chlorophyll (a+b) expressed as a concentration - Total carotenoids - amount of carotenoids in sample - Total carotenoids (micro mol.g/dw) - carotenoids expressed as a concentration - Carotenoid:Chlorophyll - carotenoids to total chlorophyll (a+b) ratio - Chlorophyll:Carotenoid - total chlorophyll (a+b) to carotenoids ratio The worksheet labelled 'Graphs Pigment Bryum' contains graphs showing means and standard error for total chlorophyll (a+b) (micro mol.g/dw), total carotenoids (micro mol.g/dw) and carotenoid:chlorophyll ratio for B. pseudotriquetrum on day 28. The worksheet labelled 'Graphs Pigment Schistidium' contains graphs showing means and standard error for total chlorophyll (a+b) (micro mol.g/dw), total carotenoids (micro mol.g/dw) and carotenoid:chlorophyll ratio for S. antarctici on day 28. The worksheet labelled 'Graphs Pigment Ceratodon' contains graphs showing means and standard error for total chlorophyll (a+b) (micro mol.g/dw), total carotenoids (micro mol.g/dw) and carotenoid:chlorophyll ratio for C. purpureus on day 28. The worksheet labelled 'Graphs Pigment Local Ceratodon' contains graphs showing means and standard error for total chlorophyll (a+b) (micro mol.g/dw), total carotenoids (micro mol.g/dw) and carotenoid:chlorophyll ratio for Local C. purpureus on day 28. The worksheet labelled 'Graphs Pigment Prasiola' contains graphs showing means and standard error for total chlorophyll (a+b) (micro mol.g/dw), total carotenoids (micro mol.g/dw) and carotenoid:chlorophyll ratio for P. crispa on day 28. References Environment Canada (2005). Biological test method: test for measuring emergence and growth of terrestrial plants exposed to contaminants in soil. Environment Canada, Method Development and Applications section, Environmental Science and Technology centre, Ontario Canada. EPS //RM/45.
Metadata ID: ASAC_1313_Transects_2002-03 Title: Windmill Islands vegetation transects, surveyed 2002/03 (baseline) This record contains data associated with a Windmill Islands vegetation baseline survey conducted in 2002/03, under ASAC_1313. A previous pilot survey is described under metadata ID: ASAC_1087_Transects_1999-00 and surveys conducted after the baseline described here are recorded in metadata IDs: ASAC_1313_Transects_2007-08, ASAC_3042_Transects_2010-11, AAS_3129_Transects_2011-12, AAS_4046_Transects_2012-13. A description of the survey design (relevant to all survey periods and above mentioned metadata IDs) follow. In 2002/03 a series of vegetation transects were established at two Windmill Island sites: (1) Antarctic Specially Protected Area (ASPA) 135, on Bailey Peninsula (66.283 S, 110.533 E), and (2) Robinson Ridge (66.368 S, 110.587 E). See SCAR Map Catalogue (maps 14450 and 14451) for location details. In summary: Each site included 10 transects. Transects spanned a vegetation community gradient, from pure bryophyte stands to the point at which the bryophyte turf was predominantly moribund and encrusted with lichens. Transect locations established in 2002/03 were in different locations within the two sites from that surveyed in 1999/2000, but remained fixed thereafter. Transect lengths ranged between approximately 1 and 4 m (for 1999/2000 pilot; length information not known for transects established 2002/03). Transects were surveyed using quadrats (portable metal frames) to mark the survey area. Three quadrat positions were established per transect: end positions referred to as "Bryophyte" and "Lichen" communities and the middle position referred to as "Transitional" community. The location of quadrat positions are marked with small metal tags glued to nearby rocks. Letters on tags are: A, M and B; where A = Lichen community, M = Transitional community and B = Bryophyte community. For each quadrat location the vegetation was sampled, collecting nine small (tweezer-pinch size) vegetation samples at 10 cm intervals across a 20 x 20 cm quadrat. The vegetation surface of each quadrat was photographed (25 x 25 cm quadrat) and environmental variables such as moisture availability were determined. For details, see QuadratSpecs.txt (this metadata record) and metadata ID: AAS_4046_Transects_2012-13, data file: Transects Data Summary_2000-2013.xlsx, worksheet "Quadrat". Vegetation samples were analysed for species composition using microscopy techniques, with live bryophytes identified to species level and lichens to broad morphological groups. Quadrat photographs were analysed for broad vegetation cover patterns (e.g. % cover live moss, dead moss, crustose lichen etc). A list of all vegetation categories scored follows (with abbreviations commonly used in associated data files provided in brackets): Live bryophytes; green (Live Bryo, Live moss) Moribund bryophytes; dead/brown/encrusted (Mori, Moribund) bryophyte species Bryum pseudotriquetrum (Bry, Bryum) bryophyte species Ceratodon purpureus (Cerat, Ceratodon) bryophyte species Schistidium antarctici (Schistidium and Grim, Grimmia*) bryophyte species Cephaloziella varians (Ceph) Fruticose Lichens (Fr) Foliose Lichens (Fo) Crustose Lichens (Cr) (* in 2002/03 this species was still referred to as Grimmia antarctici, abbreviation: Grim, thereafter it is referred to as Schistidium antarctici). Further details for transect sample collection are provided in Ryan-Colton 2007, King 2009 and Benny 2013 (and for pilot survey conducted in 1999/2000, see Wasley 2004 and Wasley et al 202). Descriptions of data associated with this record are provided below under the following headings: 1. LOCATION (GPS) DATA (and MAPS) 2. QUADRAT PHOTOS 3. NOTEBOOK SCANS 4. MICROSCOPY SCORE SHEETS 5. FINESCALE SPECIES ABUNDANCE (MICROSCOPY) 6. BROADSCALE PERCENT COVER (IMAGE ANALYSIS) 7. ENVIRONMENTAL VARIABLES (e.g. MOISTURE, TEMPERATURE) 8. PROCESSED/COMPILED/WORKED Descriptions of data provided: 1. LOCATION (GPS) DATA (and MAPS) See data provided in metadata ID: AAS_4046_Transects_2012-13 and maps 14450 and 14451 in the SCAR Map Catalogue: http://data.aad.gov.au/aadc/mapcat/display_map.cfm?map_id=14450 and http://data.aad.gov.au/aadc/mapcat/display_map.cfm?map_id=14451. 2. QUADRAT PHOTOS TO BE PROVIDED - photo files for all quadrats (and transect/site images) 3. NOTEBOOK SCANS TO BE PROVIDED - as scanned PDFs - Laboratory notebook - Jane Wasley has hardcopy (to scan) at AAD - Field notebook - Jane Wasley has hardcopy (to scan) at AAD 4. MICROSCOPY SCORE SHEETS FILE: ASAC_1313-Transects 2002-03-Microscopy.pdf The provided pdf file is a scanned copy of A4 pages that were used as score sheets for microscopy analysis of vegetation samples collected from transects in 2002/03. Samples were analysed (for species composition) via microscopy techniques after collection (analysis date/s not known, except 20/2/03 which is given on the last page of the provided file (page 20 of 20) is 20/2/03). These samples will have been analysed over a long period of time (weeks?) around this date. One page per transect. Transects located at two sites: "Meltpuddle" and "Robbos" (corresponding to M and R in quadrat labels, respectively). Transect and quadrat locations are as provided in metadata ID: Windmill Islands Vegetation Transects (noting that Meltpuddle = ASPA 135). Each transect includes three quadrats (labelled: A, M, B). A = lichen community, M = transitional community, B = bryophyte community Description of terms and abbreviations: - Transect: Transect ID; two sites: Meltpuddle and Robbos, numbers 1-10 are transect number - Quadrat: Quadrat ID; e.g. M2A (USNEA). First character = site (M=Meltpuddle; R= Robbos), second character = transect number (1-10), third character = community (A=Lichen, M=Transitional, B=Bryopyte) - Tray: label and position information of where the nine samples per quadrat were stored. Samples were stored in tissue culture trays with wells (24 wells per tray; to be confirmed via lab or field notebooks) e.g. 17 (1-3) would indicate tray number 17 (rows, 1-3). - Sample ID: e.g. 1A, 3C. Description of vegetation categories scored: Bryophytes were determined as dead (brown) or live (green). If live then identified to species. Lichens were determined as macro or crustose lichens. If macro then identified to taxa. Full list of vegetation categories follows, terms provided in brackets ( ) are those used in metadata ID: Windmill Islands Vegetation Transects, if different from that used here: BRYOPHYTES - - Dead Bryophytes: bryophytes; dead/brown/encrusted (Moribund) - Live Brophytes: bryophytes; live/green (Live Bryo) Bryum = bryophyte species Bryum pseudotriquetrum Ceratodon = bryophyte species Ceratodon purpureus Grimmia = bryophyte species Grimmia Antarctici (Schistidium antarctici) Ceph = bryophyte species Cephaloziella varians LICHENS - - Macro Lichens: Usnea = lichen taxa Usnea spp. (Fruticose Lichens) Umbilicaria = lichen taxa Umbilicaria sp. (Foliose Lichens) Pseudophebe = lichen taxa Pseudophebe sp. (Fruticose Lichens) - Crustose Lichen: lichen taxa, all species of crustose form (Crustose Lichens) Hand written scores (numbers 0-4) are observations via microscopy analysis of each tweezer-pinch sized vegetation sample. Each sample was scored for relative abundance of each vegetation category, using a modified Braun- Blanquet scale (dominant=4, co-dominant=3, low abundance=2, trace=1 and absent=0). Details as described in Wasley et al 2012, Wasley 2004 and Ryan-Colton 2007. Hand written notes include: - Apotheca: fungal fruiting bodies obseved - NSUS: No sample collected, Usnea spp. present at sampling point - NSRO: No sample collected, rock at sampling point - NSCR: No sample collected, crustose lichens at sampling point - NSPS: No sample collected, Pseudophebe spp. present at sampling point - NSPS: No sample collected, Umbilicaria spp. present at sampling point - +US: Usnea spp. present at sampling point, in addition to the sample collected. - +US (CD): Usnea spp. co-dominant at sampling point (in addition to the sample collected). Note: this file was previously named: Transects 1999-2000_microscopy score sheets.pdf - indicating the data relate to samples collected in 1999/2000. Jane Wasley has changed this to 2002/03 (July 2015) for the following reasons: - the date on the last page (page 20 of 20) is 20/2/03 - Each transect only has three quadrats (A,M,B). This was the experimental design followed in 2002/3 (onward), but in the 1999/2000 survey there up to 17 quadrats per transect. This file has therefore been renamed 2002/3. Further information should be checked via raw data and field/lab notebooks and can be updated if required. 5. FINESCALE SPECIES ABUNDANCE (MICROSCOPY) FILE: Transects-Microscopy-2002-03.xls Note, this file (prior to 2015) had been located at: SOE_Windmill_Island_veg.zip\SOE_Windmill_Island_veg\2008 Data updates\Transect Microscopy.xls, indicating it was 2007/08 data. In July 2015, Jane Wasley renamed the file as 2002/03. Other sources should be crossed check to confirm this change from 2008 to 2003 is correct (JW thinks 2008 was the year this data was submitted to AADC, but the data relates to samples collected in 2002/03). Data provided in worksheet "raw" are values between 0-4, as described in worksheet "metadata". Samples were scored for species/taxa relative abundance using a modified Braun- Blanquet scale (dominant=4, co-dominant=3, low abundance=2, trace=1 and absent=0). This scoring method was also used for the 1999/2000 survey (metadata ID: ASAC_1087_Transects_1999-00), as described in Wasley et al 2012 and Wasley 2004. Field names as per other descriptions related to this metadata record (and other related "transects" metadata records). Some details provided in worksheet "metadata". 6. BROADSCALE PERCENT COVER (IMAGE ANALYSIS) FILE: Transects-%Cover-2002-03.xls Note, as per previous file, this file (prior to 2015) had been located at: SOE_Windmill_Island_veg.zip\SOE_Windmill_Island_veg\2008 Data updates\Transect % cover.xls, indicating it was 2007/08 data. In July 2015, Jane Wasley has renamed the file as 2002/03. The file provided is raw present cover data, collected in the field via visual observations (by Jane Wasley). Quadrat area = 20 x 20 cm, gridded at 5 cm intervals. Each quadrat has 16 observations recorded (labelled using a row/column system, labelled A1 through to D4; each representing one 5 x 5 cm grid square). Note: this methods was also used for 1999/2000 transects, but was not included from 2007/08 onward - replaced instead by analysis of quadrat photos. Worksheet names: Raw Field Book - raw data as recorded in field notebook %A - working step one, calculation of percent cover from raw data %B - working step two, calculation of percent cover from raw data Metadata - descriptions of abbreviations and terms. 7. ENVIRONMENTAL VARIABLES (e.g. MOISTURE, TEMPERATURE) FILE: Transects-TurfWaterContent-2002-03.xls Note, as per previous files, this file (prior to 2015) had been located at: SOE_Windmill_Island_veg.zip\SOE_Windmill_Island_veg\2008 Data updates\Turf Water Content.xls, indicating it was 2007/08 data. In July 2015, Jane Wasley has renamed the file as 2002/03. The file provided is raw turf water content data, measured via deployment of sponge cores into moss turf. Sponge cores were inserted into the vegetation, left to equilibrate for a period of time then collected in pre-weighed air tight tubes and returned to the laboratory. Wet weight was recorded (g). Sponges dried to determine dry weight (g). These data used to calculate TWC. TWC = (Wet weight-Dry weight)/Dry weight (g. H2O g-1 dw). Two worksheets: 'raw'contains data and 'metadata'provides descriptions for data field names and abbreviations. 8. PROCESSED/COMPILED/WORKED FILE: SoE72_update_0108.doc This word doc provides a summary of survey status. Likely prepared by Ellen Ryan-Colton, but details not known. Date prepared unknown. Appears to provide data collected 2002/03 only. FILES: Ellen Ryan-Colton 2007_Thesis.pdf and Ellen Ryan-Colton 2007_Appendix.pdf The files provided are PDF files of the University of Wollongong Honours thesis written by Ellen Ryan-Colon in 2007. This thesis presents the 2002/03 baseline survey data, provides a toolkit detailing survey methods for future surveys and trials some image analysis techniques (as an alternative to collecting broadscale percent cover data visually in the field). Data were collected from ASPA 135 and Robinson Ridge, as shown in maps 14450 and 14451 in the SCAR Map Catalogue.