Detection and genetic characterization of Mamastrovirus 5 from Brazilian dogs

Alves, Christian D.B.T.; Budaszewski, Renata F.; Torikachvili, Marcela; Streck, André F.; Weber, Matheus N.; Cibulski, Samuel P.; Ravazzolo, Ana P.; Lunge, Vagner R.; Canal, Cláudio W.

A total of 269 dog fecal samples were collected between 2008 and 2014 in veterinary clinics and hospitals by convenience. These samples were obtained from eight Federal States of Brazil (Acre, Mato Grosso do Sul, Paraná, Rio Grande do Sul, Rio de Janeiro, Rondônia, Santa Catarina and São Paulo). The animal's age was recorded and ranked from puppy (equal or less than one-year-old) to adult dog (more than one-year-old); some samples from dogs of unknown age were included. Animals not presenting diarrhea at the time of sampling were considered asymptomatic and those presenting clinical signs of enteric disease diarrhea were classified as symptomatic. Samples were diluted to 20% (w/v) in phosphate buffered saline (PBS, pH 7.4) and stored at −80 • C for further analysis. Subsequently, viral DNA isolation from the supernatant was performed using a commercial kit (NewGene Preamp ® , Simbios Biotecnologia, Brazil) based on guanidine isothiocyanate and silica. 27 Viral RNA was isolated using TRIzol ® LS Reagent (Life Technologies TM , USA) according to the manufacturer's instructions.