Isolation and Molecular Characterization of herpesvirus from Argentinean water buffaloes

SILVINA SOLEDAD MAIDANA; JOSÉ LUIS KONRAD; ETIENNE TRIRY; GUSTAVO CRUDELI; SONIA ALEJANDRA ROMERA

Puket

Congreso; 10th Congreso Mundial de Búfalos y 7th Congreso de Búfalo Asiático; 2013

Asociación internacional de búfalos

BoHV-1 is distributed worldwide and is endemic in Argentina, where it is responsible for important economic losses due to the decreased production. However few are known about the seroprevalence and circulation of herpesviruses in other ruminant species. Water buffalo breeding represents an important economic alternative in Argentina, which allows access to national and international markets. Therefore is necessary to evaluate the circulation of economically important respiratory virus in buffalo herds. Within this framework, the study of viral agents present in buffalos becomes relevant. The studies of herpesviruses that circulate in these species and find out the role of buffalos as reservoirs and the possible existence of recombinant viruses with an unknown virulence are interesting. We proposed to analyze the circulating of field isolates of herpesvirus from bubaline herds in Argentina. Our previous results showed herpesvirus seroprevalence in herds of buffaloes of 34% in our country with this evidence we decided to expand the sampling of nasal and genital secretions to detect the viral agent. Nasal and genital swabs buffalos without symptoms were evaluated. The viruses were isolated from vaginal secretions from three different farms in Corrientes province of Argentina. The isolates were amplified in MDBK cell culture and characterized by indirect immunofluorescence assay (IFA) and restriction endonuclease analysis (REA) with BstEII enzyme. After viral DNA extracted was digested with the enzyme then underwent an electrophoretic run in 0.7% agar gel finally stained with ethidium bromide and the banding pattern was observed with UV light. A gene fragment coding for the glycoprotein B was amplified by PCR and sequenced. Then phylogenetic analysis was performed based on this genomic fragment using neighborjoining method and the Kimura 2-parameter model the software MEGA 4.0. The monoclonal against herpesvirus bovine was able to detect the cells infected with these viruses by IFA. The isolates showed a different restriction profile between them with enzyme analyzed. On the other hand one of them showed a similar profile to the reference strain of buffalo herpesvirus (B6) and the others two isolates showed a different pattern of buffalo reference strain and bovine herpesvirus reference strains. Phylogenetic analysis shows that two of the isolates grouped directly with the bovine herpesvirus 5 from Argentine and the remaining isolate groups directly with reference buffalo herpesvirus. In the present study we report three new isolates which added to the two previously reported for us showing the circulations of herpesvirus at least in two argentine provinces. These data constitute a contribution to the characterization of alphaherpesvirus strains circulating in ruminants in Argentina.