Hall, J.S. Nashold, S. Hofmeister, E. Leon, A.E. Falendyzsz, E.A. Ip, H.S. Malavé, C.M. Rocke, T.E. Carossino, M. Balasuriya, U. Knowles, S. 20240829 spreadsheet ScienceBase U.S. Geological Survey data release https://doi.org/10.5066/P13R5WEG Little brown bat (Myotis lucifugus) weights and SARS-CoV2 test results were collected as part of study to assess transmission potential of SARS-CoV2 in North American bat populations. It has been proposed that the SARS-CoV-2 virus originated in Asian bats and subsequently spread through human populations as a pandemic. There is concern that infected humans could transmit the virus to native North American bats, therefore the susceptibility of several North American bat species to the pandemic virus has been experimentally assessed. Big brown bats (Eptesicus fuscus) were shown to be resistant to infection by SARS-CoV-2, while Mexican free-tailed bats (Tadarida brasiliensis) became infected and orally excreted moderate amounts of virus for up to 18 days post-inoculation. Little brown bats (Myotis lucifugus) frequently contact humans, and their populations are threatened over much of their range due to white-nose syndrome, a fungal disease that is continuing to spread across North America. For this study, we experimentally challenged little brown bats with SARS-CoV-2 to determine their susceptibility, host potential, and whether the virus presents an additional risk to this species. We present data, including oral and rectal excretion, health status and serological evidence that shows this species was resistant to infection by SARS-CoV-2. These findings will provide reassurance to wildlife rehabilitators, biologists, conservation scientists, and the public at large who are concerned with possible transmission of this virus to threatened bat populations. To determine the impact of SARS-CoV2 on little brown bat populations. 20211001 20211130 ground condition Complete None planned US National Wildlife Health Center -89.485660 -89.483299 43.050325 43.047691 ISO 19115 Topic Category biota None SARS-CoV-2 little brown bat Myotis lucifugus challenge infection susceptibility resistance USGS Thesaurus polymerase chain reaction histology pathology wildlife disease zoonotic diseases USGS Metadata Identifier USGS:664506b9d34e1955f5a42f6d None National Wildlife Health Center (NWHC), Madison, Wi None Myotis lucifugus Kingdom Animalia Subkingdom Bilateria Infrakingdom Deuterostomia Phylum Chordata Subphylum Vertebrata Infraphylum Gnathostomata Superclass Tetrapoda Class Mammalia Subclass Theria Infraclass Eutheria Order Chiroptera Suborder Yangochiroptera Superfamily Vespertilionoidea Family Vespertilionidae Subfamily Myotinae Genus Myotis Subgenus Myotis (Pizonyx) Species Myotis lucifugus TSN: 179988 None. Please see 'Distribution Info' for details. None. Users are advised to read the dataset's metadata thoroughly to understand appropriate use and data limitations. Jeffrey Hall U.S. Geological Survey National Wildlife Health Center Research Virologist mailing and physical

6006 Schroeder Rd

Madison WI 53711 USA 608-270-2458 csv Jefferey Hall Sean Nashold Erik Hofmeister Ariel E Leon Elizabeth A Falendysz Hon S Ip Carly M Malavé Tonie E Rocke Mariano Carossino Udeni Balasuriya Susan Knowles 20240726 publication The Journal of Wildlife Diseases (JWD) Wildlife Disease Association (WDA) Calibrated scale produced weights. yes provides laboratory verified bat weights and PCR Ct values n/a n/a Animal monitoring and sampling Bats were observed at least twice daily to document development of clinical signs and to monitor health status. Prior to inoculation, and every other day thereafter, each bat was weighed, and oropharyngeal and rectal swabs (Puritan Medical Products, Guilford, ME) were collected and placed in 0.5 mL VTM. On day post-inoculation (DPI) 7 and on DPI 14, bats from one cage (one inoculated bat, one uninoculated) were euthanized, a postmortem examination was conducted, and tissues and blood collected. At the end of the study (DPI 20), all remaining bats were euthanized, and the control bats and an additional cage pair had postmortem examinations conducted. All euthanized bats had blood collected for serological analyses. Quantitative RT-PCR (qRT-PCR) analyses RNA extractions of swab material were performed in 96-well plates using Mag Max-96 AI/ND Viral RNA Isolation Kit (Applied Biosystems, Foster City, CA) following the manufacturer’s instructions. A positive control sample consisting of a 1:100 dilution of the SARS-CoV-2 inoculum used in the study was included with each extraction series to validate successful RNA extraction. qRT-PCR analyses were conducted utilizing the Centers for Disease Control 2019-nCoV N1 primers and probe (https://www.cdc.gov/coronavirus/2019-ncov/lab/rt-pcr-panel-primer-probes.html) and AgPath-ID One-Step RT-PCR reagents (Ambion/ThermoFisher, Waltham, MA). We included a standard curve of serial dilutions of RNA extracted from SARS-CoV-2 virus stock (107 TCID50/ml) in each qRT-PCR assay to quantify viral amounts. 20211130 Hall_littlebrownbat_weights_SARS-CoV2.csv Comma Separated Value (CSV) file containing data. Producer Defined Bat ID Unique, lab-assigned identifier for each bat Producer Defined 612 8491 Treatment Dosage type: Inoculated, Transmission, and Control Producer Defined Control Not inoculated with virus Producer defined Inoculated Inoculated with virus Producer defined Transmission Exposed to inoculated bats Producer defined Day 0 Day post inoculation. 0 = day of inoculation Producer Defined 7.9 11.2 grams DPI 2 Day post-inoculation. 2 days after inoculation Producer Defined 8.3 11.2 grams DPI 4 Day post-inoculation. 4 days after inoculation Producer Defined 8.1 11.5 grams DPI 6 Day post-inoculation. 6 days after inoculation Producer Defined 7.5 11.2 grams DPI 8 Day post-inoculation. 8 days after inoculation Producer Defined X No weight taken Producer defined 7.9 11.4 grams DPI10 Day post-inoculation. 10 days after inoculation Producer Defined x no weight taken Producer defined 7.9 11.5 grams DPI12 Day post-inoculation. 12 days after inoculation Producer Defined x weight not taken Producer defined 7.2 10.9 DPI14 Day post-inoculation. 14 days after inoculation Producer Defined x weight not taken Producer defined 7.5 11 grams DPI16 Day post-inoculation. 16 days after inoculation Producer Defined x weight not taken Producer defined 7.7 10.7 grams DPI18 Day post-inoculation. 18 days after inoculation Producer Defined x weight not taken Producer defined 7.1 10.3 grams DPI20 Day post-inoculation. 20 days after inoculation Producer Defined x weight not taken Producer defined 7.4 10.2 Hall_littlebrownbat_Ct_SARS-CoV2.csv Comma Separated Value (CSV) file containing data. Producer Defined Bat ID Unique, lab-assigned identifier for each bat Producer Defined 612 8491 Treatment Dosage type: Inoculated, Transmission, and Control Producer Defined Control Not inoculated with virus Producer defined Inoculated Inoculated with virus Producer defined Transmission. Exposed to inoculated bats Producer defined Day 0 Day post inoculation. 0 = day of inoculation Producer Defined No Ct No Ct value was generated from testing Producer defined Values provided are the Cycle threshhold (Ct) values. DPI 2 Day post-inoculation. 2 days after inoculation Producer Defined No Ct No Ct value was generated from testing Producer defined Values provided are the Cycle threshhold (Ct) values. DPI 4 Day post-inoculation. 4 days after inoculation Producer Defined No Ct No Ct value was generated from testing Producer defined Values provided are the Cycle threshhold (Ct) values. DPI 6 Day post-inoculation. 6 days after inoculation Producer Defined No Ct No Ct value was generated from testing Producer defined Values provided are the Cycle threshhold (Ct) values. DPI 8 Day post-inoculation. 8 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI10 Day post-inoculation. 10 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI12 Day post-inoculation. 12 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI14 Day post-inoculation. 14 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI16 Day post-inoculation. 16 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI18 Day post-inoculation. 18 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. DPI20 Day post-inoculation. 20 days after inoculation Producer Defined No Ct, X No Ct = Values provided are the Cycle threshhold (Ct) values. X = No testing performed Producer defined Values provided are the Cycle threshhold (Ct) values. 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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/P13R5WEG None 20240829 Jeffrey Hall U.S. Geological Survey National Wildlife Health Center Research Virologist mailing and physical

6006 Schroeder Rd

Madison WI 53711 USA 608-270-2458 jshall@usgs.gov FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata FGDC-STD-001.1-1999