engleski

Detection, isolation and partial sequencing of bovine herpesvirus-1 from an outbreak of infectious bovine rhinotracheitis

In the present study, we report the detection and isolation of bovine herpesvirus type 1 (BHV-1) in nasal swabs samples and tissue samples derived from naturally infected cattle. Clinical symptoms from the respiratory tract were observed in one dairy herd. Sera and nasal swabs from 14 animals were collected at 5th and 27th after appearance of symptoms. Tissue sample (lungs) from 8 weeks old calf was also collected. Specific antibodies against BHV-1 were not found in sera samples collected after 1st sampling, but all sera samples were positive after second sampling. The supernatants from nasal swabs dipped in Eagle’ s MEM containing antibiotics were used for virus isolation in the bovine embryo kidney (BEK) cells and for viral DNA extraction. The cytopathyc effect was visible after 72 hours for the nasal swabs samples, and after 48 hours for the lung sample. Sensitive PCR assays specific for glycoprotein B (gB) and gC were also performed. All nasal swab samples and a lung sample were positive with both sets of PCR primers. Obtained gC PCR product from the lung sample DNA was sequenced. Phylogenetic analysis of nucleotide and aminoacid gC sequence clustered our isolate with BHV subtype 1.1. The results of laboratory testing confirmed the BHV-1 to be the causative agent of respiratory tract disease observed in the cattle herd.

infectious bovine rhinotracheitis; bovine herpesvirus-1; Croatia

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