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ARS Home » Midwest Area » Madison, Wisconsin » U.S. Dairy Forage Research Center » Environmentally Integrated Dairy Management Research » Research » Publications at this Location » Publication #418109
Research Project: Managing Nutrients and Assessing Pathogen Emission Risks for Sustainable Dairy Production Systems

Location: Environmentally Integrated Dairy Management Research

Title: Validation of laboratory recovery for human and swine pathogens from two types of ultrafilters

Author
item DOUGHAN, GABI - Iowa State University
item FIRNSTAHL, AARON - Us Geological Survey (USGS)
item Opelt, Sarah
item Cook, Rachel
item Heffron, Joseph
item KRUEGER, KAREN - Iowa State University
item Borchardt, Mark
item KARRIKER, LOCKE - Iowa State University
item STOKDYK, JOEL - Us Geological Survey (USGS)
item Burch, Tucker

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 8/26/2024
Publication Date: 9/10/2024
Citation: Doughan, G., Firnstahl, A.D., Opelt, S.A., Cook, R.M., Heffron, J.A., Krueger, K.M., Borchardt, M.A., Karriker, L.A., Stokdyk, J.P., Burch, T.R. 2024. Validation of laboratory recovery for human and swine pathogens from two types of ultrafilters. Meeting Abstract. September 10-11, 2024.

Interpretive Summary:

Technical Abstract: Dead-end ultrafiltration is a well-established mechanism for collecting and concentrating multiple types of pathogens from water samples with many pathogen types validated. The Asaha Kasei Rexeed ™ hemodialyzer is the most widely used ultrafilter for dead-end ultrafiltration, but has become unavailable in the U.S. The Nipro Elisio ™ ultrafilters have been identified as a potential commercially available replacement but have not yet been validated to have comparable recovery efficiencies. Additionally, there has been an increased interest from veterinarians to use dead-end ultrafiltration for pathogen detection in livestock water sources, thus motivating further validation for swine pathogens of interest: porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2). The objectives of this study were to compare the recovery efficiency between the Asaha Kasei Rexeed ™ and Nipro Elisio ™ hemodialyzers and demonstrate recovery of swine pathogens PRRSV and PCV2. Ten liters of dechlorinated tap water were spiked with target organisms (PRRSV-2, PCV2, Salmonella enterica serovar Typhimurium, adenovirus 41, Campylobacter jejuni, Cryptosporidium parvum, and poliovirus Sabin type 3). Spiked water was then concentrated via either Asahi Kasei Rexeed-25A ™ (n=6) or Nipro Elisio-25H ™ ultrafilters (n=6). Six control samples were also spiked with target organisms into backflush solution, avoiding the filtration step. Test and control samples underwent polyethylene glycol precipitation, nucleic acid extraction, reverse transcription (for RNA viruses), and were then tested via qPCR. Primary recovery and full recovery efficiencies were calculated for each microorganism. PRRSV-2 and PCV2 were recovered via qPCR by each filter type from spiked water samples. PRRSV was recovered via qPCR from all filters, with average primary recovery efficiencies of 64–78%. PCV2 was recovered in 4 of 12 filters, 2 from each filter type, indicating recovery was successful for PCV2, but concentrations were near detection limits. Average primary recovery efficiencies for target pathogens were similar between filter types. Average recovery efficiencies for bacteria ranged from 33% to 76% (Rexeed™) and 39% to 84% (Elisio™). Average recovery efficiencies for viruses ranged from 17% to 64% (Rexeed™) and 10% to 78% (Elisio™). Average recovery efficiencies for Cryptosporidium parvum were 98% (Rexeed™) and 100% (Elisio™). Results demonstrate that one can concentrate a variety of pathogens, including swine pathogens of interest (PRRSV-2 and PCV2), with either Rexeed-25A or Elisio-25H filters with comparable recovery efficiencies.