Wendy C. Turner Zoe R. Barandongo 20230111 spreadsheet https://doi.org/10.5066/P9TDAY91 Datasets collected from monitored zebra anthrax carcass sites in Etosha National Park, Namibia, 2010-2021. One data file describes the anthrax mortality, soil characterization of the carcass sites and details on the types and timing of different sampling efforts at these sites. The other data file is counts of Bacillus anthracis spores in surface soils collected annually at these carcass sites. Data were collected for disease surveillance efforts and research into anthrax transmission ecology. 20100202 20211231 observed Complete None planned central Etosha National Park, Namibia 15.6995 16.0936 -18.9842 -19.2697 USGS Metadata Identifier USGS:63bc141ed34e92aad3cd2b12 ISO 19115 Topic Category biota geoscientificInformation USGS Thesaurus wildlife disease Common geographic areas Namibia None Equus quagga burchellii Bacillus anthracis Life Life Kingdom Animalia Subkingdom Bilateria Infrakingdom Deuterostomia Phylum Chordata Subphylum Vertebrata Infraphylum Gnathostomata Superclass Tetrapoda Class Mammalia Subclass Theria Infraclass Eutheria Order Perissodactyla Family Equidae Genus Equus Subgenus Equus (Hippotigris) Species Equus quagga Subspecies Equus quagga burchellii TSN: 926246 Kingdom Bacteria Subkingdom Posibacteria Phylum Firmicutes Class Bacilli Order Bacillales Family Bacillaceae Genus Bacillus Species Bacillus anthracis TSN: 959822 Precision of GPS location information was reduced for biosecurity considerations. None. Users are advised to read the dataset's metadata thoroughly to understand appropriate use and data limitations. Wendy Turner Wisconsin Cooperative Wildlife Research Unit mailing and physical

1630 Linden Drive

Madison WI 53706 608-263-6882 wturner@usgs.gov Etosha Ecological Institute Zoe R. Barandongo 20230228 DNA sequences https://www.ncbi.nlm.nih.gov/sra/PRJNA914838 Spore count data are estimated from 10-fold serial dilutions, and hence should not be considered precise. The diagnoses of anthrax are accurate and confirmed with microbiological and genetic tests. Animal age data are estimates based on tooth emergence and incisor wear patterns based off Smuts (1974; https://journals.co.za/doi/10.10520/AJA03794369_2679). The date of mortality is the date the mortality was recorded, not the date of death. Estimates of the date of death ranged from 0-11 days prior to sampling with a mean of 1 day before the mortality was recorded. Values fall within expected ranges. No data are duplicated or omitted. This dataset is complete for these zebra anthrax carcass sites that were monitored over a 12 year period. NA NA Bacillus anthracis spore counts: Soil samples were collected at the soil surface (0-1cm depth) in the center of the carcass site, with many subsamples within 1m of the marked center of the carcass site collected with a sterile spoon pooled into one sample for analyses. These soil samples were assessed via bacterial culture in serial dilution on PLET (polymyxin B sulfate, lysozyme, EDTA and thallous acetate) agar, methods adapted from standard protocols (see paper associated with this dataset). For the soil samples, five grams of homogenized soil was combined with 45ml 0.1% sodium pyrophosphate in a 50ml centrifuge tube then vortexed at maximum speed for 10 minutes to break up soil clumps. Samples were then centrifuged at 0.3x1000 rcf for 2 minutes to settle larger soil particles and the supernatant transferred to a new centrifuge tube and centrifuged at 3.0 x 1000 rcf for 15 minutes to pellet the sample. The pellet was resuspended in 5ml of sterile distilled water. One ml of the resuspended pellet was used to create serial dilutions from 100 to 10-5 (the number of dilutions plated depended on the site age and sample type), and 100μl of the selected dilutions was plated directly. Plates were placed in an incubator at 30C for two days. Colonies of B. anthracis were identified morphologically, and plates were examined and B. anthracis colonies counted after two days. In cases where the identification of a colony was uncertain, the colony was streaked on nutrient agar for B. anthracis confirmation tests to determine if the colony had penicillin sensitivity and was lysed by gamma phage. The number of CFU/g was determined for all dilutions that were countable. Quantification of the CFU/g of soil was based on the plate with the smallest dilution in the dilution series that had CFU counts of 30-300 colonies, or the lowest dilution with colonies, if all counts were smaller than 30. 20211231 Soil characterization: Soil samples from each carcass site (four 10cm cores, pooled by site) were collected near carcass sites to test if soil characteristics altered Bacillus anthracis (BA) persistence over time, measuring soil texture and composition, pH, organic matter, P, K, Ca, Mg, and Na. These cores were collected in each direction 9m from the marking stake at the center of the carcass site. This is because carcasses can significantly alter soils and we were interested in local soil conditions without the effect of the carcass. One sample was collected per site in 2012/2013 and a second in 2014. These two samples were analyzed independently but were similar enough that we averaged the two samples and the data reported here are the average of these two samples from each site. Soil analyses were conducted at the Ministry of Agriculture, Water and Forestry Laboratory in Windhoek, Namibia using standard methods (Association of Official Analytical Chemists, Official Methods of Analysis of the AOAC International, 1996). 20161231 carcass_site_description.csv Comma Separated Value (CSV) file containing data. Producer Defined mortality_ID identifying label from mortality surveillance Producer Defined This ID includes a 2-letter species code, date (YYMMDD) - the number of mortalities of that species detected on that day by the observer, and a 2-letter code for the observer's initials carcass_ID identifying label of blood swab for laboratory purposes Producer Defined A number containing the year and swab number per year (YY-###). species species of the recorded mortality (all plains zebras for this study) Producer Defined Equus quagga plains zebra Producer defined sex sex of recorded mortality Producer Defined NA No Data Producer defined female the animal mortality was female Producer defined male the animal mortality was male Producer defined age_category age categorized as adult (>2yrs) or subadult (1-2 years) old, or as described in mortality records Producer Defined adult adult (2+ years old) Producer defined subadult subadult (1-2 years old) Producer defined estimated_age estimated age or age range of the animal mortality in years, based on tooth emergence and incisor wear patterns Producer Defined NA No Data Producer defined 6 6 years Producer defined 5 5 years Producer defined NA No Data Producer defined 11-13 11-13 years Producer defined 5-6 5-6 years Producer defined 10 10 years Producer defined 11 11 years Producer defined 16 16 years Producer defined 4-5 4-5 years Producer defined 6-7 6-7 years Producer defined 12 12 years Producer defined 2.5-3 2.5-3 years Producer defined 1.5-2 1.5-2 years Producer defined 3-4 3-4 years Producer defined 1.5-3 1.5-3 years Producer defined 3 3 years Producer defined 8-9 8-9 years Producer defined 7 7 years Producer defined date_mortality date the mortality was recorded. Typically the animal is estimated to have died within the prior 24-48 hours. Producer Defined << empty cell >> No Data Producer defined Recorded in DD-Mon-YY cause_death cause of death Producer Defined anthrax All mortalities in this sample were confirmed anthrax positive based on microbiological testing Producer defined month_of_death the month of death in numbers Producer Defined 2 8 week_of_death the week of death (possible range 1-52) Producer Defined << empty cell >> No Data Producer defined 6 36 season season of death Producer Defined wet January-April Producer defined dry May-October Producer defined longitude longitude of the mortality location Producer Defined << empty cell >> No Data Producer defined 15.727 16.049 decimal degrees latitude latitude of mortality location Producer Defined << empty cell >> No Data Producer defined -19.231 -18.994 decimal degrees isolate_ID identifying number of a bacterial isolate collected from each carcass or carcass site Producer Defined Each isolate has a number in the format of WT-## SRA_accession NCBI SRA accession number for DNA sequences from the bacterial isolate Producer Defined NA No Data Producer defined Isolate was not sequenced or sequence quality did not meet quality control standards isolate_source sample source from which the Bacillus anthracis strain was isolated Producer Defined Soil collected in 2016 Bacteria isolated from surface soil from the annual soil sample that was collected in 2016. Producer defined Blood swab collected in 2010 Bacteria isolated from the stored swab collected when the carcass was first recorded. Producer defined Blood swab collected in 2011 Bacteria isolated from the stored swab collected when the carcass was first recorded. Producer defined Soil collected in 2017 Bacteria isolated from surface soil from the annual soil sample that was collected in 2017. Producer defined Soil collected in 2020 Bacteria isolated from surface soil from the annual soil sample that was collected in 2020. Producer defined Soil collected in 2018 Bacteria isolated from surface soil from the annual soil sample that was collected in 2018. Producer defined isolation_month-year the month/year when the bacterial strain was isolated Producer Defined May-16 May 2016 Producer defined Jul-17 July 2017 Producer defined Jul-18 July 2018 Producer defined Jun-19 June 2019 Producer defined Jun-20 June 2020 Producer defined DNA_extraction_month-year the month/year when DNA was extracted from the bacterial isolate Producer Defined Jul-17 July 2017 Producer defined Jul-19 July 2019 Producer defined Jan-18 January 2018 Producer defined Aug-18 August 2018 Producer defined Jul-17 July 2017 Producer defined Aug-19 August 2019 Producer defined Jun-20 June 2020 Producer defined soil_texture the texture of the soil at the carcass site based on USDA classification Producer Defined Loam Loam Producer defined Sandy loam Sandy loam Producer defined Silty loam Silty loam Producer defined sand the percentage of sand in the soil sample Producer Defined 34.3 76.3 percent silt the percentage of silt in the soil sample Producer Defined 20.1 58.6 percent clay the percentage of clay in the soil sample Producer Defined 3.6 18.7 percent om the percentage of organic matter in the soil sample Producer Defined << empty cell >> No Data Producer defined 1.15 2.655 percent P Amount of phosphorus in the soil sample Producer Defined << empty cell >> No Data Producer defined 1.25 8.775 ppm K Amount of potassium in the soil sample Producer Defined << empty cell >> No Data Producer defined 409.5 3401.0 ppm Ca Amount of calcium in the soil sample Producer Defined << empty cell >> No Data Producer defined 2922.0 5114.5 ppm Mg Amount of magnesium in the soil sample Producer Defined << empty cell >> No Data Producer defined 209.0 1019.0 ppm Na Amount of sodium in the soil sample Producer Defined << empty cell >> No Data Producer defined 29.0 1255.0 ppm pH pH of the soil sample Producer Defined << empty cell >> No Data Producer defined 8.29 9.01 spore_count_data.csv Comma Separated Value (CSV) file containing data. Producer Defined carcass_ID identifying label of the animal mortality for laboratory purposes Producer Defined A number containing the year and swab number per year (YY-###). sample_date sampling date Producer Defined The date a soil sample was collected. Age_yrs carcass site age in years after death when soil sample collected Producer Defined 0 11 years Age_days carcass site age in days after estimated death when soil sample collected Producer Defined 11 4165 days CFU colony forming units of Bacillus anthracis, an estimate of the number of spores per gram of soil Producer Defined 0 1193044852 U.S. Geological Survey GS ScienceBase mailing address

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Denver CO 80225 United States 1-888-275-8747 sciencebase@usgs.gov Unless otherwise stated, all data, metadata and related materials are considered to satisfy the quality standards relative to the purpose for which the data were collected. Although these data and associated metadata have been reviewed for accuracy and completeness and approved for release by the U.S. Geological Survey (USGS), no warranty expressed or implied is made regarding the display or utility of the data for other purposes, nor on all computer systems, nor shall the act of distribution constitute any such warranty. Digital Data https://doi.org/10.5066/P9TDAY91 None 20230111 Wendy Turner Wisconsin Cooperative WIldlife Research Unit mailing and physical

1630 Linden Drive

Madison WI 53706 608-263-6882 wturner@usgs.gov FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata FGDC-STD-001.1-1999