ABSTRACT
Abstract Malignant Catarrhal Fever (MCF) was investigated in the central nervous system of cattle with neurological syndrome. Two-hundred-ninety samples were analyzed by histology, and molecular methods to detect ovine herpesvirus type 2 (OvHV-2) were optimized and validated. The qualitative polymerase chain reaction (qualitative PCR) analytical sensitivity was 101 DNA copies/µL and found 4.8% (14/290) positive for OvHV-2. The quantitative polymerase chain reaction (qPCR) analytical sensitivity was 100 DNA copy/µL and 5.9% (17/290) positivity, with 47.1% (8/17) of the positive samples presenting histological evidence of non-purulent meningo-encephalitis. The qualitative PCR products (422 bp of the ORF75 region) were sequenced and submitted to phylogenetic analysis. Identity matrices showed 100% similarity in OvHV-2 samples obtained in this study and those recovered from GenBank, corroborating other studies.
Subject(s)
Animals , Phylogeny , Molecular Diagnostic Techniques/methods , Herpesviridae/isolation & purification , Malignant Catarrh/diagnosis , Malignant Catarrh/pathology , Brazil , Cattle , Cluster Analysis , Polymerase Chain Reaction/methods , Sensitivity and Specificity , Sequence Homology , Sequence Analysis, DNA , Genotype , Herpesviridae/classification , Herpesviridae/genetics , Histocytochemistry , MicroscopyABSTRACT
An outbreak of Malignant Catarrhal Fever (MCF) resulted in death of five female buffaloes and one domestic cow from the same farm. Four buffaloes died 10-15 days after the appearance of clinical signs, while the fifth was euthanized in extremis, after similar clinical signs. Histopathological lesions included multifocal histiolymphocytic epicarditis, myocarditis and lymphocytic interstitial pneumonia, which are commonly seen in cases of MCF in buffaloes. Furthermore, lymphocytic vasculitis centered in the adventitia, with occasional fibrinoid necrosis in the muscular layer, was found in the kidneys, liver, spleen, lymph nodes and brain. Nucleotide sequencing of DNA fragments from the central nervous system amplified by PCR revealed 98 percent similarity with known OHV-2 sequences from Genbank. Additionally, PCR analysis also revealed the presence of OHV-2 DNA in the peripheral mononuclear blood cells of two clinically healthy buffaloes. The diagnosis of MCFwas based on epidemiological, clinical, gross and histopathological findings and on the results of a semi-nested PCR followed by nucleotide sequencing.
É relatado um surto de febre catarral maligna (FCM) em Minas Gerais, que resultou na morte de 5 búfalas e uma vaca de uma mesma propriedade. Quatro búfalas morreram com 10-15 dias após o início dos sinais clínicos e uma búfala foi sacrificada in extremis, após manifestar sinais clínicos semelhantes. O exame histopatológico revelou lesões comumente observadas em búfalos com FCM como epicardite e miocardite histiolinfocítica multifocal e pneumonia linfocítica intersticial. Além disso, vasculite linfocítica, principalmente na camada adventícia, com necrose fibrinóide da camada muscular, foi observada no rim, fígado, baço, linfonodos e cérebro. A seqüência de nucleotídeos amplificada pela técnica de PCR revelou 98 por cento de homologia entre o fragmento de DNA amplificado da amostra do sistema nervoso central (SNC) da búfala com seqüências de OHV-2 previamente depositadas no Genbank. Adicionalmente, a técnica de PCR revelou a presença do DNA viral no sangue total de 2 búfalas, clinicamente sadias. O diagnóstico de FCM foi baseado em dados epidemiológicos, clínicos, patológicos, histopatológicos e semi-nested PCR.