title: Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil.
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.
authors.affiliation: Laboratório de Virologia Aplicada, Departamento de Microbiologia, Imunologia e Parasitologia, UFSC, Florianópolis, Brazil.
authors.forename: Gislaine
authors.initials: G
authors.lastname: Fongaro
authors.affiliation: Instituto Tecnológico Agrario de Castilla y León, Valladolid, Spain.|Departamento de Ingeniería Agrícola y Forestal, Tecnología de los Alimentos, E.T.S. Ingenierías Agrarias, Universidad de Valladolid, 34004, Palencia, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Hernández
authors.affiliation: Instituto Tecnológico Agrario de Castilla y León, Valladolid, Spain.
authors.forename: María Cruz
authors.initials: MC
authors.lastname: García-González
authors.affiliation: Laboratório de Virologia Aplicada, Departamento de Microbiologia, Imunologia e Parasitologia, UFSC, Florianópolis, Brazil.
authors.forename: Célia Regina Monte
authors.initials: CR
authors.lastname: Barardi
authors.affiliation: Microbiology Section, Department of Biotechnology and Food Science, Universidad de Burgos, Burgos, Spain. rodlazda@gmail.com.
authors.forename: David
authors.initials: D
authors.lastname: Rodríguez-Lázaro
mesh_terms: D000818:Animals; D001386:Azides / Q000737:chemistry; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D005308:Fertilizers / Q000032:analysis; D008372:Manure / Q000821:virology*; D011419:Propidium / Q000031:analogs & derivatives / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012987:Soil / Q000737:chemistry; D012988:Soil Microbiology; D013552:Swine; D014774:Virulence
chemical_list: D001386:Azides; D005308:Fertilizers; D008372:Manure; D012987:Soil; C533957:propidium monoazide; D011419:Propidium
keywords: Biofertilizer; Infectious enteric viruses; PMA–PCR; Recycle of swine manure