Jenna L. K. Shelton Madalyn S. Blondes Varun Saraswathula Elliott P. Barnhart Colin A. Doolan Jessica C. McKinley Eric A. Morrissey 20210825 Data presented are raw data, including the blank sample, with no contaminants removed. These data will be used as such in the related publication that will proceed this data release. Reston, VA U.S. Geological Survey https://doi.org/10.5066/P92RJOJS In order to determine the innate microbial community of shale gas reservoirs and how they are impacted by hydraulic fracturing, this study analyzed biomass collected from produced water and rock from hydraulically fractured wells in the Utica Shale. The samples include rock chips from a drill core from one Utica well, produced water from that same Utica well, and produced water from 12 different Utica wells that had been in production between 1-5 years, spanning the oil and gas windows of SE Ohio. The samples were filtered for biomass, extracted, amplified, and 16S rRNA gene sequencing was performed on the Illumina MiSeq platform. The data were collected because minimal data exist for the microbiology of the Utica Shale. We also wanted to determine if there were microbiological differences between formation rock and formation water, as well as determine if any changes to microbial community composition could occur over time. 20170802 20170803 The time represents when the water was filtered for microbial analysis. Complete None planned -82.0 -81.0 40.5 39.5 Utica Shale wells in southeastern Ohio USGS Thesaurus Field Sampling Time Series Microbial Ecology 16S rRNA Gene Sequencing Shale Hydraulic Fracturing Utica Shale Produced Water USGS Metadata Identifier USGS:5b0ea0fde4b0c39c934b132e Geographic Names Information System Southeastern Ohio Utica Shale Geolex - U.S. Geologic Names Lexicon None USGS Thesaurus None none none Jenna L Shelton NORTHEAST REGION: GEOLOGY, ENERGY&MINERALS SC Research Hydrologist mailing and physical

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Sacramento CA 95819 US 916-278-3221 jlshelton@usgs.gov Map showing sample locations jpeg Argonne National Laboratory performed all upstream analyses, including DNA extraction, polymerase chain reaction, and Illumina MiSeq 16S rRNA sequencing. USGS Biocomplexity Thesaurus Microorganisms Pelin Yilmaz Laura Wegener Parfrey Pablo Yarza Jan Gerken Elmar Pruesse Christian Quast Timmy Schweer Jörg Peplies Wolfgang Ludwig Frank Oliver Glöckner 2014 ONLINE_REFERENCE Unknown Nucleic Acids Research https://academic.oup.com/nar/article/42/D1/D643/1061236 Jenna L Shelton NORTHEAST REGION: EASTERN ENERGY RESOURCES SC Research Hydrologist mailing and physical

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Sacramento CA 95819 US 916-278-3221 jlshelton@usgs.gov genetic analysis The sequences identified were mapped to OTUs at >99% sequence similarity and were mapped to the species level when possible. Seventeen water samples (including duplicates) and 2 rock samples were collected for microbial biomass. The Archaea identified are species of the Phyla Euryarchaeota and Thaumarchaeota. The Bacteria identified are species of the Phyla Acidobacteria; Actinobacteria; Armatimonadetes; Bacteroidetes; Candidate Divisions JBRC1, JL-ETNP-Z39, KB1, NPL-UPA2, OD1, OP3, SHA-109, TA06, TM6, TM7, WS3; Chlamydiae; Chlorobi; Chloroflexi; Cyanobacteria; Fibrobacteres; Firmicutes; Fusobacteria; Gemmatimonadetes; Lentisphaerae; Nitrospirae; Planctomycetes, Proteobacteria; Spirochaetae; Synergistetes; Tenericutes; Thermotogae; and Verrucomicrobia. Two Eukaryotic OTU were detected from the Phylums Opisthokonta and SAR. Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom,Kingdom Unassigned,Eukaryota,Eukaryota,Eukaryota,Eukaryota,Eukaryota,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Bacteria,Archaea Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum,Phylum Unclassified,SAR,SAR,Opisthokonta,Opisthokonta,Excavata,Verrucomicrobia,Verrucomicrobia,Verrucomicrobia,Verrucomicrobia,Verrucomicrobia,Verrucomicrobia,Verrucomicrobia,Thermotogae,Tenericutes,TM6,TM6,TA06,Synergistetes,Spirochaetae,SHA-109,Proteobacteria,Proteobacteria,Proteobacteria,Proteobacteria,Proteobacteria,Proteobacteria,Proteobacteria,Planctomycetes,Planctomycetes,Planctomycetes,Planctomycetes,Planctomycetes,Planctomycetes,Planctomycetes,Planctomycetes,Nitrospirae,NPL-UPA2,Lentisphaerae,Lentisphaerae,Lentisphaerae,JL-ETNP-Z39,Gemmatimonadetes,Fusobacteria,Firmicutes,Firmicutes,Firmicutes,Firmicutes,Firmicutes,Firbrobacteres,Cyanobacteria,Cyanobacteria,Cyanobacteria,Chloroflexi,Chloroflexi,Chloroflexi,Chloroflexi,Chloroflexi,Euryarchaeota Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class,Class Unclassified,Stramenopiles,Alveolata,Nucletmycea,Holozoa,Metamonada,Verrucomicrobiae,Verrucomicrobia Incertae Sedis,Spartobacteria,S-BQ2-57 soil group,Opitutae,OPB35 soil group,Arctic97B-4 marine group,Thermotogae,Mollicutes,Unclassified,uncultured bacterium,uncultured bacterium,Synergistia,Spirochaetes,uncultured bacterium,Unclassified,SC3-20,Gammaproteobacteria,Epsilonproteobacteria,Deltaproteobacteria,Betaproteobacteria,Alphaproteobacteria,Unclassified,vadinHA49,Planctomycetacia,Pla4 lineage ,Pla3 lineage ,Phycisphaerae,OM190,BD7-11,Nitrospira,uncultured Omnitrophia bacterium,WCHB1-25,RFP12 gut group,Lentisphaeria,uncultured bacterium,Gemmatimonadetes,Fusobacteriia,OPB54,Negativicutes,Erysipelotrichia,Clostridia,Bacilli,Fibrobacteria,Melainabacteria,ML635J-21,Cyanobacteria,Unclassified,Thermomicrobia,TK10,S085,Ktedonobacteria,Halobacteria Kingdom Archaea Phylum Euryarchaeota Class Methanobacteria Kingdom Archaea Phylum Euryarchaeota Class Thermoplasmata Kingdom Archaea Phylum Thaumarchaeota Class Soil Crenarchaeota Group (SCG) Kingdom Bacteria Phylum Acidobacteria Class Acidobacteria Kingdom Bacteria Phylum Acidobacteria Class Holophagae Kingdom Bacteria Phylum Acidobacteria Class Subgroup 22 Kingdom Bacteria Phylum Actinobacteria Class Acidimicrobiia Kingdom Bacteria Phylum Actinobacteria Class Actinobacteria Kingdom Bacteria Phylum Actinobacteria Class Coriobacteriia Kingdom Bacteria Phylum Actinobacteria Class MB-A2-108 Kingdom Bacteria Phylum Actinobacteria Class Rubrobacteria Kingdom Bacteria Phylum Actinobacteria Class TakashiAC-B11 Kingdom Bacteria Phylum Actinobacteria Class Thermoleophilia Kingdom Bacteria Phylum Actinobacteria Class Unassigned Kingdom Bacteria Phylum Armatimonadetes Class Chtonomonadetes Kingdom Bacteria Phylum Armatimonadetes Class Uncultured Armatimonadetes bacterium Kingdom Bacteria Phylum Armatimonadetes Class Uncultured bacterium Kingdom Bacteria Phylum Armatimonadetes Class Uncultured soil bacterium Kingdom Bacteria Phylum Armatimonadetes Class Unassigned Kingdom Bacteria Phylum Bacteroidetes Class Bacteroidia Kingdom Bacteria Phylum Bacteroidetes Class Cytophagia Kingdom Bacteria Phylum Bacteroidetes Class Flavobacteriia Kingdom Bacteria Phylum Bacteroidetes Class Sphingobacteriia Kingdom Bacteria Phylum Bacteroidetes Class VC2.1 Bac22 Kingdom Bacteria Phylum Bacteroidetes Class Unclassified Kingdom Bacteria Phylum Candidate Division BRC1 Class uncultured bacterium Kingdom Bacteria Phylum Candidate Division KB1 Class uncultured Chloroflexi bacterium Kingdom Bacteria Phylum Candidate Division OD1 Class uncultured bacterium Kingdom Bacteria Phylum Candidate Division OD1 Class uncultured organism Kingdom Bacteria Phylum Candidate Division OD1 Class uncultured soil bacterium Kingdom Bacteria Phylum Candidate Division OD1 Class Unclassified Kingdom Bacteria Phylum Candidate Division OP3 Class uncultured bacterium Kingdom Bacteria Phylum Candidate Division TM7 Class Unknown Class Kingdom Bacteria Phylum Candidate Division TM7 Class Unassigned Kingdom Bacteria Phylum Candidate Division WS3 Class uncultured Acidobacteria bacterium Kingdom Bacteria Phylum Candidate Division WS3 Class uncultured soil bacterium Kingdom Bacteria Phylum Candidate Division WS3 Class uncultured bacterium Kingdom Bacteria Phylum Candidate Division WS3 Class uncultured prokaryote Kingdom Bacteria Phylum Candidate Division WS3 Class Unclassified Kingdom Bacteria Phylum Chlamydiae Class Chlamydiae Kingdom Bacteria Phylum Chlorobi Class Chlorobia Kingdom Bacteria Phylum Chlorobi Class Ignovibacteria Kingdom Bacteria Phylum Chloroflexi Class Anaerolineae Kingdom Bacteria Phylum Chloroflexi Class Ardenticatenia Kingdom Bacteria Phylum Chloroflexi Class Caldilineae Kingdom Bacteria Phylum Chloroflexi Class Chloroflexia Kingdom Bacteria Phylum Chloroflexi Class Dehalococcoidia Kingdom Bacteria Phylum Chloroflexi Class Gitt-GS-136 Kingdom Bacteria Phylum Chloroflexi Class KD4-96 These data were processed using QIIME2 and various packages in the program R (R Core Team, 2017). The QIIME2 tool/program allowed for the raw, unmerged, chimeric sequences to be cleaned, mapped to OTUs and produced as a Feature Table (e.g., an OTU table). After the OTU table was produced, R was used to further clean the data, remove contaminants, and rarefy the dataset. The packages used were the base packages as well as vegan, reshape2, and biom. https://docs.qiime2.org/2020.2/install/ Visit https://docs.qiime2.org/2020.2/install/ and install QIIME2. Follow the "Moving Pictures" tutorial to process data. Jenna L Shelton NORTHEAST REGION: GEOLOGY, ENERGY&MINERALS SC Research Hydrologist mailing and physical

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Sacramento CA 95819 US 916-278-3221 jlshelton@usgs.gov Caporaso, J.G., et al. 2010 View No formal attribute accuracy tests were conducted No formal logical accuracy tests were conducted. Data set is considered complete for the information presented, as described in the abstract. Users are advised to read the rest of the metadata record carefully for additional details. No formal positional accuracy tests were conducted. No formal positional accuracy tests were conducted. Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO 20160929 .tgz file Online European Nucleotide Archive https://www.arb-silva.de/documentation/release-132 nucleotide sequencing information 20200422 20200425 https://www.arb-silva.de/ Silva v132 Database The input data mapped the identified sequences to known operational taxonomic units (OTUs) QIIME2 was used to process the raw sequence reads into a feature table. The steps performed are outlined below. Read in as single reads, not paired reads due to low quality of reverse reads limiting sequences per sample. $qiime tools import \ --type EMPPairedEndSequences \ --input-path emp-paired-end-sequences \ --output-path emp-paired-end-sequences.qza $ qiime demux emp-paired \ --m-barcodes-file mapping.txt \ --m-barcodes-category BarcodeSequence \ --i-seqs emp-paired-end-sequences.qza \ --o-per-sample-sequences demux.qza \ --p-rev-comp-mapping-barcodes \ --p-rev-comp-barcodes $ qiime demux summarize \ --i-data demux.qza \ --o-visualization demux.qzv $qiime dada2 denoise-paired \ --i-demultiplexed-seqs demux.qza \ --p-trunc-len-f 250 \ --p-trunc-len-r 215 \ --trim-left-f 12 \ --trim-left-r 12 \ --o-representative-sequences rep-seqs.qza --o-table table.qza $qiime feature-table summarize \ --i-table table.qza \ --o-visualization table.qzv \ --m-sample-metadata-file mapping.txt $ qiime feature-table tabulate-seqs \ --i-data rep-seqs.qza \ --o-visualization rep-seqs.qzv $qiime feature-classifier classify-sklearn --i-classifier silva-119-99-515-806-nb-classifier.qza \ --i-reads rep.seqs.qza --p-reads-per-batch 4000 --o-classification taxonomy.qza $qiime taxa collapse / --i-table table.qza / --i-taxonomy taxonomy_silva70conf.qza / --p-level 7 / --o-collapsed-table OTU_table_silva70conf.qza $qiime tools export / OTU_table_silva70conf.qza / --output-dir exported-OTU-table_silva70conf $biom convert / -i exported-OTU-table_silva70conf/feature-table.biom / -o OTU-table.tsv / --to-tsv 04242020 Jenna L Shelton NORTHEAST REGION: GEOLOGY, ENERGY&MINERALS SC Research Hydrologist mailing and physical

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Sacramento CA 95819 US 916-278-3221 jlshelton@usgs.gov both Samples were collected from various locations and from both water and rock for this study. The rock core chips (n=4) were collected from a well in the Utica Shale, B3H, that was being drilled for hydraulic fracturing. A water sample (n=1) was also collected from this newly-drilled well, B3H. The remaining water samples (n=16) were collected from various wells across Ohio, all of which had been hydraulically fractured. One blank was also included. To collect biomass samples, formation water (stored in a separator tank) was collected in a sterile 2-gallon plastic carboy that had been washed three times with sample water. This water was filtered using sterile Nalgene tubing and a GeoPump through Sterivex 0.22 um filter units. Duplicates were collected from many wells, while one blank sample was also collected by pumping 2 L of Milli-Q through a Sterivex filter unit. After sample collection, the filters were immediately placed on dry ice, shipped to the Eastern Energy Resources Science Center (United States Geological Survey), and placed into a -80 °C freezer. The samples that came from well B3H (the rock core chips and one water sample) were collected slightly differently. The rock was collected from a well that was being drilled for hydraulic fracturing. That rock was collected in a ziploc bag and shipped to the Eastern Energy Resources Science Center (USGS) and placed into a -80 °C freezer until it was sampled. Once the well was drilled and started producing water, a sample of that water was collected in a clean, sterile, rinsed plastic carboy and shipped to the Eastern Energy Resources Science Center (USGS). Once it arrived, it was placed in a refrigerator until it could be filtered. The water was filtered using the same methods as described above, except the sample water was poured into a clean glass beaker (and one end of the Nalgene tubing was placed into the beaker) for filtering. In order to extract DNA from the rock chips, the outside faces of the rock were removed using a sterile chisel, and the inside portion was crushed into a fine sediment. The DNA from the sediment was extracted using the MO BIO PowerSoil DNA Extraction kit following the manufacturer’s protocol, at Argonne National Laboratory. The DNA from all Sterivex™ filters was extracted using the MO BIO PowerWater DNA Extraction kit following the manufacturer’s protocol at Argonne National Laboratory. The V4 region of the 16S rRNA gene (515F-806R) was PCR amplified with region-specific primers that include the sequencer-adapter sequences used in the Illumina flowcell. The forward amplification primer also contains a twelve base barcode sequence that supports the pooling of samples in each lane. Each 25 μL PCR reaction contains 9.5 μL of MO BIO PCR Water (Certified DNA-Free), 12.5 μL of QuantaBio’s AccuStart II PCR ToughMix (2x concentration, 1x final), 1μL Golay barcode tagged forward primer (5 μM concentration, 200 pM final), 1 μL reverse primer (5 μM concentration, 200 pM final), and 1 μL of template DNA. The conditions for PCR were as follows: 94 °C for 3 minutes to denature the DNA, with 35 cycles at 94 °C for 45 seconds, 50 °C for 60 seconds, and 72 °C for 90 seconds, with a final extension of 10 minutes at 72 °C to ensure complete amplification. Amplicons are then quantified using PicoGreen (invitrogen) and a plate reader (Infinite 200 PRO, Tecan). Once quantified, volumes of each of the products are pooled into a single tube so that the amplicon is represented in equimolar amounts (for comparison across samples). This pool is then cleaned up using AMPure XP Beads (Beckman Coulter), and then quantified using a fluorometer (Qubit, Invitrogen). After quantification, the molarity of the pool is determined and diluted down to 2 nM, denatured, and then diluted to a final concentration of 6.75 pM with a 10% PhiX spike for sequencing on the Illumina MiSeq. DNA sequence data were generated using Illumina MiSeq paired-end sequencing at the Environmental Sample Preparation and Sequencing Facility (ESPSF) at Argonne National Laboratory using 251 bp x 12 bp x 251 bp chemistry and customized sequencing primers and procedures. QIIME2 Version 2020.2 was used to demultiplex the raw data, merge and align the forward and reverse sequences, perform quality control, and assign taxonomy based on the Silva version 132 taxonomic classifier. The procedures outlined in the “Atacama Soil Microbiome” and the “Moving Pictures” tutorials on the QIIME2 website (https://docs.qiime2.org/2020.2/tutorials/) were followed, with some modifications. The reads were trimmed so that the primers were removed (12 base pairs; --p-trim-left-f 12\ --p-trim-left-r 12), and were truncated at the position when the average quality score dropped below 30 (--p-trunc-len-f 250\ --p-trunc-len-r 215). The generated Feature Table (OTU table with taxonomy) was then outputted from QIIME2 and loaded into R (R Core Team, 2017). U.S. Geological Survey 2017 Book 9, chaps. A1-A10 http://pubs.water.usgs.gov/twri9A All latitude and longitude coordinates were collected. Point 1.0E-4 1.0E-4 Decimal degrees NAD83 GRS80 6378137.0 298.257222101 OTU Table.csv The spreadsheet contains the sample names/locations (column A), and the OTUs identified (columns B through EAB), and the number of times each OTU was identified in each sample collected. The Sample IDs are the field-derived names of the sampled locations, and the OTU numbers were created by the Silva v132 database. The number of times each OTU was identified depends on the microbiology of the sample collected. These data were collected in the field, and provided in raw form by Argonne National Laboratory (Lemont, IL, USA). Sample ID The producer defined names for the samples. Producer defined The field contains field defined names for the samples. OTU"X" An alpha numeric identifier related to the operational taxonomic unit identified. Each identifier is unique and corresponds to an assigned organism in the Silva version 132 database. The "X" refers to a number, 1 through 3407. The actual taxonomic identifications are provided in a separate entity (Taxonomic Table Utica 042420.csv). Silva version 132 database 0 24103 Counts, or number of times the given operational taxonomic unit was observed across the dataset Tax Table.csv The spreadsheet contains the names of the operational taxonomic units (OTUs) from the Entity "OTU Table Utica 042420.csv" in column A, and the specific names, given the taxonomic classification (e.g., kingdom, phylum, etc.), assigned to that OTU number. The taxonomic classifications (e.g., species name) were assigned by the Silva v132 Database. Assigned OTU An alphanumeric identifier tied to a specific microorganism found in the Silva version 132 database. Silva version 132 database Silva version 132 Database https://www.arb-silva.de/documentation/release-132/ Kingdom The taxonomic Kingdom that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Phylum The taxonomic Phylum that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Class The taxonomic Class that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Order The taxonomic Order that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Family The taxonomic Family that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Genus The taxonomic Genus that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ Species The taxonomic Species that the given OTU is assigned to. National Center for Biotechnology Information All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ All attribute values can be identified in the "Taxonomy" Database on the National Center for Biotechnology (NCBI) website, https://www.ncbi.nlm.nih.gov/guide/taxonomy/ The sequence reads identified were mapped to particular Operational Taxonomic Units described in the Silva v132 database. The Operational Taxonomic Units identified in this project are unique to the study area and are only those observed in the study area during the sampling of water from each location (only those observed during the observation period). Therefore, only some Operational Taxonomic Units described in the Silva v132 database were observed here, as not all microorganisms can live in every environment on Earth. All OTUXXX numbers can be found in the silva database, and all Kingdom, Phylum, Class, Order, Family, Genus, and Species names can be located and described on the National Center for Biotechnology (NCBI) website. The Silva database defines the column headers in the OTU Table entity, while the NCBI database describes the attributes under each column header in the second entity, the Taxonomy Table. https://www.arb-silva.de/documentation/release-132/ https://www.ncbi.nlm.nih.gov/ Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucl. Acids Res. 41 (D1): D590-D596. https://academic.oup.com/nar/article/41/D1/D590/1069277 Yilmaz P, Parfrey LW, Yarza P, Gerken J, Pruesse E, Quast C, Schweer T, Peplies J, Ludwig W, Glöckner FO (2014) The SILVA and "All-species Living Tree Project (LTP)" taxonomic frameworks. Opens external link in new windowNucl. Acids Res. 42:D643-D648. https://academic.oup.com/nar/article/42/D1/D643/1061236 GS ScienceBase U.S. Geological Survey mailing and physical

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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. The file contains data available in comma separated value (.csv) file format. The user must have software capable of opening and viewing a .csv file. 20210825 Eric A Morrissey NORTHEAST REGION: GEOLOGY, ENERGY&MINERALS SC Info Tech Spec (Internet) mailing and physical

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Reston VA 20192 US 703-648-6409 emorriss@usgs.gov FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata FGDC-STD-001.1-1999