Laboratory assessment of recovery of porcine circovirus 2 and porcine reproductive and respiratory syndrome virus using 2 commercial hollow-fiber ultrafilters

Groundwater near swine farms is an uninvestigated reservoir for porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCVs). Enteric microorganisms are often collected from groundwater via dead-end ultrafiltration, but recovery of PRRSV and PCV with this method has not been assessed. We recovered PRRSV2 and PCV2 by dead-end ultrafiltration followed by polyethylene glycol (PEG) precipitation, nucleic acid extraction, and reverse-transcription quantitative real-time PCR. We also compared 2 commercial hemodialysis ultrafilters (Asahi Kasei Rexeed-25A, Nipro Elisio-25H) and compared PRRSV2 recovery in these filters to other waterborne microorganisms. On average, 8 ± 1% of PRRSV2 was recovered by dead-end ultrafiltration and PEG precipitation, compared to 25 ± 6% for adenovirus 41. Full-process recovery of bacteria in the same filters was 5-15%; <i>Cryptosporidium parvum</i> recovery was 42 ± 12%. PCV2 was detected in 4 of 12 replicate filters, but low stock concentrations precluded quantitative recovery estimates. Elisio-25H ultrafilters performed similarly to Rexeed-25A filters for all organisms tested and is an effective replacement for the Rexeed-25A, which is no longer available in the United States. Our recovery of PRRSV2 and PCV2 by dead-end ultrafiltration in the laboratory suggests that PRRSV2 detection limits are as low as 3-50 genomic copies/L in sample volumes of 100-1,500 L. Based on quantitative microbial risk assessment, these concentrations are relevant to PRRSV2 infection rates in the U.S. swine herd.

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Publication Details

Journal
Journal of Veterinary Diagnostic Investigation
Published
2025-02-28
DOI
https://doi.org/10.1177/10406387251322506
Citations
1
Primary Topic
Animal Virus Infections Studies
Type
article
Field-Weighted Citation Impact
1.19

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article

Laboratory assessment of recovery of porcine circovirus 2 and porcine reproductive and respiratory syndrome virus using 2 commercial hollow-fiber ultrafilters

Karen Krueger, Mark A. Borchardt, Gabrielle Doughan, Joel P. Stokdyk et al.
1 citations
Journal of Veterinary Diagnostic Investigation
Animal Virus Infections Studies
1.19
article

Laboratory assessment of recovery of porcine circovirus 2 and porcine reproductive and respiratory syndrome virus using 2 commercial hollow-fiber ultrafilters

Karen Krueger, Mark A. Borchardt, Gabrielle Doughan, Joel P. Stokdyk, Rachel M. Cook, Sarah A. Opelt, Aaron D. Firnstahl, Tucker R. Burch, Locke A. Karriker, Joseph A. Heffron
article en
1 citations

Abstract

Groundwater near swine farms is an uninvestigated reservoir for porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCVs). Enteric microorganisms are often collected from groundwater via dead-end ultrafiltration, but recovery of PRRSV and PCV with this method has not been assessed. We recovered PRRSV2 and PCV2 by dead-end ultrafiltration followed by polyethylene glycol (PEG) precipitation, nucleic acid extraction, and reverse-transcription quantitative real-time PCR. We also compared 2 commercial hemodialysis ultrafilters (Asahi Kasei Rexeed-25A, Nipro Elisio-25H) and compared PRRSV2 recovery in these filters to other waterborne microorganisms. On average, 8 ± 1% of PRRSV2 was recovered by dead-end ultrafiltration and PEG precipitation, compared to 25 ± 6% for adenovirus 41. Full-process recovery of bacteria in the same filters was 5-15%; <i>Cryptosporidium parvum</i> recovery was 42 ± 12%. PCV2 was detected in 4 of 12 replicate filters, but low stock concentrations precluded quantitative recovery estimates. Elisio-25H ultrafilters performed similarly to Rexeed-25A filters for all organisms tested and is an effective replacement for the Rexeed-25A, which is no longer available in the United States. Our recovery of PRRSV2 and PCV2 by dead-end ultrafiltration in the laboratory suggests that PRRSV2 detection limits are as low as 3-50 genomic copies/L in sample volumes of 100-1,500 L. Based on quantitative microbial risk assessment, these concentrations are relevant to PRRSV2 infection rates in the U.S. swine herd.

Journal of Veterinary Diagnostic InvestigationVol. 37(3)
United States Geological Survey (US), Agricultural Research Service (US), Iowa State University (US), Dairy Management (US), Upper Midwest Water Science Center
Agricultural Research Service
Clean water and sanitation
Openalex Percentile: Top 28%
Animal Virus Infections Studies
1.19
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