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Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil

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Abstract

The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC0), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9 %). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques.

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Acknowledgments

This study was funded by the Spanish project RTA2014-0024-C04-01 of the Spanish Ministry of Economy and Innovation and the Brazilian CNPq Project number 472804/2013-8 of the CAPES/PNPD and CAPES/PDSE.

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Authors and Affiliations

  1. Laboratório de Virologia Aplicada, Departamento de Microbiologia, Imunologia e Parasitologia, UFSC, Florianópolis, Brazil

    Gislaine Fongaro & Célia Regina Monte Barardi

  2. Instituto Tecnológico Agrario de Castilla y León, Valladolid, Spain

    Marta Hernández & María Cruz García-González

  3. Departamento de Ingeniería Agrícola y Forestal, Tecnología de los Alimentos, E.T.S. Ingenierías Agrarias, Universidad de Valladolid, 34004, Palencia, Spain

    Marta Hernández

  4. Microbiology Section, Department of Biotechnology and Food Science, Universidad de Burgos, Burgos, Spain

    David Rodríguez-Lázaro

Authors
  1. Gislaine Fongaro
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  2. Marta Hernández
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  3. María Cruz García-González
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  4. Célia Regina Monte Barardi
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  5. David Rodríguez-Lázaro
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Correspondence to David Rodríguez-Lázaro.

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Fongaro, G., Hernández, M., García-González, M.C. et al. Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil. Food Environ Virol 8, 79–85 (2016). https://doi.org/10.1007/s12560-015-9225-1

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  • DOI: https://doi.org/10.1007/s12560-015-9225-1

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