Relata-se um caso de meningoencefalite causada por Herpesvirus bovino 5 (BoHV-5) heritabilityem uma vaca com cinco anos de idade. O animal manifestou quadro clínico inicial de síndrome medular baixa, caracterizada por incoordenação dos membros pélvicos, sinais estes ainda não descritos para a enfermidade. Dentro de pouco tempo a doença evoluiu para síndrome cerebral, e o óbito ocorreu seis dias após o inicio dos sintomas. Na histopatologia, evidenciou-se meningoencefalite difusa, não supurada, e a confirmação do diagnóstico foi feita por reação em cadeia de polimerase e sequenciamento do segmento parcial da glicoproteína G do vírus. O trabalho confirma a presença do BoHV-5 em Minas Gerais, descreve características clínicas novas para a enfermidade e ressalta sua importância no diagnóstico diferencial das neuropatias bovinas.

A clinical case of meningoencephalitis by Bovine herpesvirus 5 (BoHV-5) in a five-year-old cow was reported. The disease began with low spinal cord signs, characterized by incoordination, and these symptoms had never been related to this illness before. Signs of a brain syndrome were observed and the cow died in six days. At the histopathology, a spread non-supurative meningoencephalitis was diagnosed, and the virus identification was made by PCR and partial sequence of the glycoprotein G. This study confirm the BoHV-5 presence in the State of Minas Gerais, Brazil, describes new clinic characteristics, and show the importance of the disease in the differentiate diagnosis with others bovine central nervous system affections.

The immunohistochemistry standardization for bovine viral diarrhea virus (BVDV) diagnostic was described. The formalin-fixed tissue samples from a heifer with mucosal disease were used as positive control. The validation of the first phase results was performed using samples from an aborted fetus and a calf infected with reference strains of BVDV. The best results were seen using monoclonal antibodies and a commercial kit consisting of labelled streptavidin biotin (LSAB) reagents and the diaminobenzidine (DAB) substrate-chromogen reagent. The immunohistochemistry demonstrated to be an useful method for routine diagnosis for the controll and detection of BVDV infection.

Utilizou-se uma reação em cadeia de polimerase (PCR) previamente desenvolvida para a amplificação de parte da região única longa 27 (UL27) do genoma de herpesvírus bovino 1.1 (BoHV-1), que codifica a glicoproteína B, buscando a diferenciação entre isolados de BoHV-1.1 e BoHV-5. Os produtos de PCR gerados a partir de isolados de BoHV-1.1 e BoHV-5 mostraram padrão de amplificação diferenciado em seus tamanhos moleculares. Analisando as seqüências de nucleotídeos dos produtos de PCR obtidos de dois isolados de BoHV-5, juntamente com as seqüências dos produtos de PCR obtidos de um isolado de BoHV-1.1 e de três isolados de BoHV-1.2, previamente depositados no GenBank, verificou-se que a diferença observada na amplificação se deve ao número de repetições de G-C presentes no final da região codificadora da gB, particularmente nas seqüências 5'-G(A/T)CC-3'. A análise dessas seqüências-motivo desponta como uma ferramenta auxiliar na diferenciação entre isolados de BoHV-1.1, BoHV-1.2 e BoHV-5.

A previously-developed PCR was used for the amplification of part of the UL27 region of the BoHV-1.1 genome, which codes for glycoprotein B, seeking the differentiation between BoHV-1.1 and BoHV-5 isolates. The PCR products generated from the BoHV-1 and BoHV-5 isolates showed a pattern of differentiated amplification of their molecular size. The PCR of the BoHV-5 products were sequenced and the results compared with the sequence of the BoHV-1.1 and BoHV-1.2 isolates previously deposited in the GenBank. It was verified that the difference between the PCR products is due to greater number of repetitions of G-C present at the end of the gB codifier region. The most common repeat sequences were 5'-G(A/T)CC-3'. The analysis of these repetitions was shown to be an auxiliary tool in the differentiation between BoHV1.1, BoHV-1.2 and BoHV-5 isolates.

Este estudo teve como objetivo a padronização do ensaio de imunoperoxidase em monocamada de células (IPM) para o diagnóstico etiológico da diarréia bovina a vírus (DBV). O teste foi padronizado em monocamada de cultivo primário de pulmão fetal bovino (PFB) inoculada com as amostras clássicas, citopatogênica (CP) e não citopatogênica (NCP), do vírus da DBV e testado em amostras biológicas suspeitas processadas no teste clássico de isolamento viral (IV). O método de IPM identificou o vírus da DBV, apresentando melhores resultados com a utilização do calor como agente fixador, a soroalbumina bovina a 4% em PBS como bloqueador e a revelação com o cromógeno 3-amino-9-etil-carbazol (AEC). Como anticorpos primários, tanto o anticorpo policlonal como o monoclonal forneceram bons resultados.

The aim of this study was to standardize the immunoperoxidase in cell monolayer assay (IPMA) for the etiological diagnosis of bovine viral diarrhea (BVD). The method was standardized in monolayer of primary bovine fetal lung culture inoculated with cytophatic and non-cytophatic classical strains of BVD virus and tested using samples that were considered suspected in the classical technique of viral isolation. The IPMA successfully identified BVD virus and presented better results when heat was used for fixation, BSA 4% solution in PBS was used for blocking and AEC chromogen was used for revelation. Both monoclonal and polycloral antibodies gave good results when used as primary antibodies.

This paper reports the detection of bovine herpesvirus type 5 (BoHV-5) by a specific nested PCR assay. Samples were collected from the central nervous system (CNS) of cattle from Minas Gerais and São Paulo States, Brazil. All animals died presenting neurological symptoms. Nineteen frozen CNS samples analyzed had been previously tested by fluorescence antibody test for rabies virus and showed negative results. Three paraffin-embedded brain tissue samples were examined by histopatology and the observed alterations suggested nonsuppurative meningoencephalitis. BoHV-5 was detected in five (22.7%) among 22 tested samples. The occurrence of BoHV-5 infection is reported in the Southeast region of Brazil, indicating that epidemiological studies should be carried out.