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Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities.

Onyekachukwu Henry Osemeke1, Isadora Machado1, Elisa De Conti1

  • 1Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USA.

Viruses
|January 25, 2025
PubMed
Summary

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Tongue fluid from dead piglets can be used for Porcine reproductive and respiratory syndrome virus (PRRSV) isolation. The phosphate-buffered saline (PBS) method yielded the highest virus isolation success rate, particularly with ZMAC cell lines.

Area of Science:

  • Veterinary Virology
  • Swine Disease Diagnostics

Background:

  • Porcine reproductive and respiratory syndrome virus (PRRSV) poses significant economic and health challenges to the global swine industry.
  • Effective PRRSV isolation is critical for disease surveillance, outbreak investigation, and the development of targeted vaccines and control strategies.

Purpose of the Study:

  • To evaluate the efficacy of tongue fluid (TF) collected from perinatal piglet mortalities as a source for PRRSV isolation.
  • To compare different TF collection protocols and assess the suitability of various cell lines for PRRSV isolation.

Main Methods:

  • Four tongue fluid collection methods were tested: phosphate-buffered saline (PBS), virus transportation medium (VTM), freeze-thaw, and tissue homogenates.
  • PRRSV isolation was attempted using two cell lines (ZMAC, MARC-145) and primary alveolar macrophages (PAM).
Keywords:
PCRPRRSVpostmortem tissuessurveillancetongue fluidsvirus isolation

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  • RT-qPCR was used to assess viral RNA load, and virus isolation (VI) success rates were determined for each method and cell line.

    • Main Results:

    • The PBS collection protocol demonstrated the highest virus isolation success rate (22.6%), followed by the VTM group (12.1%). Freeze-thaw and homogenate methods showed significantly lower success rates (2.8% each).
    • The ZMAC cell line yielded the highest PRRSV isolation rate (21.0%) compared to MARC-145 (3.1%) and PAM cells (4.8%).
    • Mean RT-qPCR Ct values were lowest for the PBS and VTM groups, indicating higher viral RNA concentrations.

    Conclusions:

    • Tongue fluid from perinatal piglet mortalities is a viable alternative sample source for PRRSV virus isolation.
    • The PBS collection method combined with the ZMAC cell line appears most effective for maximizing PRRSV isolation from this sample type.
    • This approach can support PRRSV monitoring and control efforts in swine populations.