Handling cultured isolates and clinical, environmental, or wildlife surveillance samples containing Risk Group 3 and 4 pathogens presents considerable biosafety challenges in minimizing human exposure during processing and transport. Safe handling typically requires high- or maximum-containment facilities, demanding substantial logistical planning and resources. We evaluated PrimeStore Molecular Transport Medium (PS-MTM), a guanidine-based solution created to kill pathogens and preserve nucleic acids at ambient temperatures, for inactivating Crimean-Congo hemorrhagic fever, eastern equine encephalitis, Ebola, Hendra, Japanese encephalitis, Lassa, Marburg, Nipah, Rift Valley fever, and West Nile viruses. To mimic diagnostic conditions, human whole blood spiked with any of these viruses was incubated with PS-MTM for 20-, 30-, or 60-min. Samples with titers up to 10⁷ PFU/mL exposed to PS-MTM at all time points resulted in complete loss of infectivity judged by plaque assays. A 30-min incubation provided a 50% safety margin over the minimum inactivation time and was used for quantification with the tissue culture infectious dose (TCID₅₀) assay, enabling evaluation of PS-MTM’s activity for viruses that do or do not produce well-defined plaques. Results confirmed that PS-MTM inactivated all tested viruses at titers up to 10⁷ TCID₅₀/mL, underscoring its reliability for enhancing biosafety in diagnostics, outbreak management, and surveillance.

Coronavirus Disease 2019 resulted in a global pandemic from 2020 to 2023. During this period various wildlife species were infected with SARS-CoV-2, most notably white-tailed deer (WTD; Odocoileus virginianus) in the United States. From 2021 through 2025, Minnesota agencies and organizations partnered to collect wildlife samples and assess the threat. By leveraging existing surveillance networks, the epidemiological role of wildlife during and after the pandemic was assessed. Peridomestic wildlife from the Wildlife Rehabilitation Center of Minnesota and various regional, state, and university disease surveillance programs were screened for SARS-CoV-2. Testing was conducted via RT-PCR, with a subset of samples also undergoing serological analysis and whole genome sequencing. From 2021 to 2025, a total of 3,478 wild animals, representing 44 species, were tested for SARS-CoV-2 by RT-PCR or serology. All animals tested negative except white-tailed deer (WTD). In 2021, 6.3% (9/142) hunter-harvested WTD were RT-PCR positive and genomically identifiable to the Delta clade of SARS-CoV-2. Thirty-eight percent (5/13) of WTD fawns were serologically positive during the same year. In 2023, among WTD, 5.6% (4/71) had RT-PCR SARS-CoV-2 detections, with 1 sample sequenced with 91.97% genome recovery to the Omicron clade, Pango lineage, typed to HF.1. No other SARS-CoV-2 detections occurred among our sample population. SARS-CoV-2 circulation within Minnesota wildlife was detected but does not appear to be sustained among wildlife populations in Minnesota. By leveraging existing partnerships and utilizing an opportunistic sampling strategy, this cross-organizational cooperation demonstrates a cost-effective model for long-term zoonotic surveillance. Ultimately, these findings underscore the necessity of a One Health framework to enhance global readiness for detecting and mitigating future spillover events.

Classical swine fever (CSF) is a highly contagious transboundary animal disease that causes major economic losses to the global swine industry and remains notifiable to the World Organisation for Animal Health (WOAH). Handling and transport of infectious classical swine fever virus (CSFV)-positive samples require biosafety level 3 (BSL-3) containment, limiting diagnostic capacity and interlaboratory exchange in many regions. In this study, we evaluated the efficacy of PrimeStore^® Molecular Transport Medium (PS-MTM) for eliminating detectable CSFV infectivity while preserving viral ribonucleic acid (RNA) detectability for downstream molecular detection. Validation assays were performed using the CSFV Alfort/187 reference strain and clinical samples collected from pigs experimentally infected with the Catalonia01 and Margarita CSFV strains. Residual infectivity was assessed by virus isolation, whereas viral RNA detection and stability were evaluated using the WOAH-recommended real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR) assay. Following PS-MTM treatment, no residual infectivity was detected in either cell culture-derived virus stocks or clinical samples, including extensive replicate testing and three subsequent blind passages. RT-qPCR analyses confirmed preservation and stability of detectable CSFV RNA for at least 60 days under the storage conditions evaluated, including refrigerated storage and room temperature storage for Catalonia01-derived samples. Overall, these findings demonstrate that PS-MTM eliminates detectable CSFV infectivity while preserving viral RNA for molecular diagnosis. The implementation of PS-MTM may facilitate safer handling, transport, and interlaboratory exchange of CSFV-positive samples, contributing to improved biosafety, diagnostic harmonization, and global surveillance capacity for this transboundary animal disease.