COMPARATIVE STUDY OF THE CLASTOGENIC ACTION OF BOVINE HERPESVIRUS 1 (BOHV-1) IN PRIMARY AND ESTABLISHED CELL CULTURES

Abstract

The concern about the transmission of infectious diseases by the use of biotechnologies, like in vitro fertilization (IVF), has been increasing. The aim of this study was to verify the clastogenic alterations in the DNA of Madin-Darby Bovine Kidney (MDBK) and Bovine Oviduct Epithelial Cells (BOEC) infected or not with bovine herpesvirus 1 (BoHV-1), for 24 hours, by the use of the Comet Assay or Single-Cell Gel Electrophoresis (SCGE). The nuclear change was evaluated by measuring the extent of DNA damage, where it was possible to observe 1.7% of level II comet and 2.1% of level IV in the BOEC group “in situ“. However, in BOEC infected by the virus the damages observed were 13.8% comet level I, 0.6% level III and 1.2% level IV. The MDBK cells not infected group showed displayed 1.9% level II and the infected group 26% level I, 12.3 % level II and 6.1 % level III. All groups, regardless if they were infected or not by the virus, were considered positive by the comet assay, maybe because there has not been virus association with the cell DNA. The number of comets detected in the BOEC “in situ” corresponds to spontaneous mutations that use to be present in organisms with failure of DNA repair.  In vitro established and primary cells show greater failure frequency in DNA repair mechanisms, and may present more mutations. Therefore, in vitro models provide better conditions to mimic  in vivo conditions. These data have demonstrated that not always the IVF problems are due to pathogens infection. DOI:  http://dx.doi.org/10.17525/vrr.v15i1.38