Subject(s)
Cattle Diseases/virology , RNA, Viral/analysis , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Bovine/isolation & purification , Animals , Base Sequence , Brazil/epidemiology , Cattle , Cattle Diseases/epidemiology , Immunohistochemistry/veterinary , Molecular Sequence Data , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Virus, Bovine/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Sequence Alignment , Seroepidemiologic StudiesABSTRACT
In a retrospective study, 51 cases of gastritis (14%) were identified from among 341 necropsies performed on simian immunodeficiency virus (SIV)-infected rhesus macaques (Macaca mulatta) at the New England Primate Research Center from 1993 to 2001. Protozoa were seen in the stomach of 13 monkeys (25%) with gastritis. Two histopathologic manifestations of gastritis were observed: seven cases of lymphoplasmacytic gastritis with trichomonad trophozoites within lumens of gastric glands and four cases of necrosuppurative gastritis containing intralesional periodic acid-Schiff-positive protozoa; two cases of gastritis had morphologic features of both types of gastritis. In instances of necrosuppurative and combined lymphoplasmacytic and necrosuppurative gastritis, protozoa were 4-35 microm in diameter and round to tear-shaped. Because of the unusual morphology of the protozoa in these latter cases, transmission electron microscopy and polymerase chain reaction (PCR) were used to further identify these organisms. The protozoa were definitively identified as Tritrichomonas in all cases on the basis of ultrastructural characteristics (flagella and undulating membranes) and amplification of a 347-bp product of the 5.8S ribosomal RNA gene of Tritrichomonas foetus, Tritrichomonas suis and Tritrichomonas mobilensis by PCR using DNA extracted from stomach tissue. On the basis of these observations, we conclude that Tritrichomonas can be a significant cofactor in the development of necrosuppurative gastritis in SIV-infected rhesus macaques.
Subject(s)
Gastritis/veterinary , Macaca mulatta , Monkey Diseases/parasitology , Monkey Diseases/virology , Protozoan Infections, Animal , Protozoan Infections/virology , Simian Acquired Immunodeficiency Syndrome/parasitology , Simian Immunodeficiency Virus/growth & development , Tritrichomonas/growth & development , Animals , DNA, Protozoan/chemistry , DNA, Protozoan/genetics , Female , Gastritis/pathology , Gastritis/virology , Immunohistochemistry/veterinary , In Situ Hybridization/veterinary , Male , Microscopy, Electron, Transmission/veterinary , Monkey Diseases/pathology , Polymerase Chain Reaction/veterinary , Protozoan Infections/parasitology , Protozoan Infections/pathology , RNA, Protozoan/chemistry , RNA, Protozoan/genetics , RNA, Ribosomal, 5.8S/chemistry , RNA, Ribosomal, 5.8S/genetics , Retrospective Studies , Simian Acquired Immunodeficiency Syndrome/pathology , Simian Acquired Immunodeficiency Syndrome/virology , Tritrichomonas/genetics , Tritrichomonas/ultrastructureABSTRACT
This paper presents the first isolation of bovine respiratory syncytial virus in Brazil and its physicochemical, morphological and molecular characterization. The virus was isolated from 33 samples of nasotracheal secretions, successively inoculated into a Madin-Darby bovine kidney cell culture, which was characterized by physicochemical tests and morphological observation by electron microscopy. The Brazilian sample is an RNA pleomorphic, enveloped, thermolabile and non-hemagglutinating spicular virus. Reverse transcription, followed by nested polymerase chain reaction (nRT-PCR) assay was carried out using oligonucleotides B1, B2A, B3 and B4 for the fusion proteins (F) and B5A, B6A, B7A and B8 for the attachment protein (G). The nRT-PCR-F amplified a fragment of 481 bp corresponding to part of the gene that codes for protein F, whereas nRT-PCR-G amplified a fragment of 371 bp, in agreement with part of the G gene. The virus isolated from Brazilian samples in this study corresponded to the bovine respiratory syncytial virus, and RT-PCR proved to be useful for the diagnosis of bovine clinical samples.
Subject(s)
Respiratory Syncytial Viruses/isolation & purification , Animals , Brazil , Cattle , Cells, Cultured , Microscopy, Electron , RNA, Viral/analysis , Respiratory Syncytial Viruses/genetics , Respiratory Syncytial Viruses/ultrastructure , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
This paper presents the first isolation of bovine respiratory syncytial virus in Brazil and its physicochemical, morphological and molecular characterization. The virus was isolated from 33 samples of nasotracheal secretions, successively inoculated into a Madin-Darby bovine kidney cell culture, which was characterized by physicochemical tests and morphological observation by electron microscopy. The Brazilian sample is an RNA pleomorphic, enveloped, thermolabile and non-hemagglutinating spicular virus. Reverse transcription, followed by nested polymerase chain reaction (nRT-PCR) assay was carried out using oligonucleotides B1, B2A, B3 and B4 for the fusion proteins (F) and B5A, B6A, B7A and B8 for the attachment protein (G). The nRT-PCR-F amplified a fragment of 481 bp corresponding to part of the gene that codes for protein F, whereas nRT-PCR-G amplified a fragment of 371 bp, in agreement with part of the G gene. The virus isolated from Brazilian samples in this study corresponded to the bovine respiratory syncytial virus, and RT-PCR proved to be useful for the diagnosis of bovine clinical samples
