ABSTRACT
Abstract Mamastrovirus 5 (MAstV5), belonging to the Astroviridae (AstV) family, previously known as canine astrovirus or astrovirus-like particles, has been reported in several countries to be associated with viral enteric disease in dogs since the 1980s. Astroviruses have been detected in fecal samples from a wide variety of mammals and birds that are associated with gastroenteritis and extra enteric manifestations. In the present study, RT-PCR was used to investigate the presence of MAstV5 in 269 dog fecal samples. MAstV5 was detected in 26% (71/269) of the samples. Interestingly, all MAstV5-positive samples derived from dogs displaying clinical signs suggestive of gastroenteritis, other enteric viruses were simultaneously detected (canine parvovirus, canine distemper virus, canine coronavirus, canine adenovirus and canine rotavirus). Based on genomic sequence analysis of MAstV5 a novel classification of the species into four genotypes, MAstV5a-MAstV5d, is proposed. Phylogenetic analyses based on the ORF2 amino acid sequences, samples described herein grouped into the putative genotype 'a' closed related with Chinese samples. Other studies are required to attempt the clinical and antigenic implications of these astrovirus genotypes in dogs.
Subject(s)
Animals , Dogs , Mamastrovirus/isolation & purification , Mamastrovirus/genetics , Astroviridae Infections/veterinary , Dog Diseases/virology , Gastroenteritis/veterinary , Phylogeny , Mamastrovirus/classification , Open Reading Frames , Astroviridae Infections/virology , Feces/virology , Gastroenteritis/virology , GenotypeABSTRACT
Mamastrovirus 5 (MAstV5), belonging to the Astroviridae (AstV) family, previously known as canine astrovirus or astrovirus-like particles, has been reported in several countries to be associated with viral enteric disease in dogs since the 1980s. Astroviruses have been detected in fecal samples from a wide variety of mammals and birds that are associated with gastroenteritis and extra enteric manifestations. In the present study, RT-PCR was used to investigate the presence of MAstV5 in 269 dog fecal samples. MAstV5 was detected in 26% (71/269) of the samples. Interestingly, all MAstV5-positive samples derived from dogs displaying clinical signs suggestive of gastroenteritis, other enteric viruses were simultaneously detected (canine parvovirus, canine distemper virus, canine coronavirus, canine adenovirus and canine rotavirus). Based on genomic sequence analysis of MAstV5 a novel classification of the species into four genotypes, MAstV5a-MAstV5d, is proposed. Phylogenetic analyses based on the ORF2 amino acid sequences, samples described herein grouped into the putative genotype 'a' closed related with Chinese samples. Other studies are required to attempt the clinical and antigenic implications of these astrovirus genotypes in dogs.
Subject(s)
Astroviridae Infections/veterinary , Dog Diseases/virology , Gastroenteritis/veterinary , Mamastrovirus/genetics , Mamastrovirus/isolation & purification , Animals , Astroviridae Infections/virology , Dogs , Feces/virology , Gastroenteritis/virology , Genotype , Mamastrovirus/classification , Open Reading Frames , PhylogenySubject(s)
Enterovirus Infections/veterinary , Enteroviruses, Porcine/isolation & purification , Lung/microbiology , Lung/virology , Pneumocystis carinii/isolation & purification , Pneumonia, Pneumocystis/veterinary , Abattoirs , Animals , Brazil/epidemiology , Enterovirus Infections/complications , Enterovirus Infections/epidemiology , Enterovirus Infections/virology , Enteroviruses, Porcine/classification , Food Microbiology , Lung/pathology , Pneumonia, Pneumocystis/complications , Pneumonia, Pneumocystis/epidemiology , Pneumonia, Pneumocystis/microbiology , Polymerase Chain Reaction , Prevalence , SwineABSTRACT
A plasmid expressing the beta-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures, with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method, we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery.
