Phylogenetic Analysis And Searching Bovine Papillomaviruses In Teat Papillomatosis Cases In Cattle By Histopathological, Immunohistochemical And Transmission Electron Microscopy Methods

Abstract

Papillomaviruses are epitheliotropic viruses causing proliferations in skin, mucosa and various internal organs in different animal species. Especially due to lesions it causes in teats of cattle, it leads to important economical losses in milk sector. In this study, the aim was to diagnose bovine papillomaviruses (BPVs) causing teat papillomas in cattle by immunohistochemical, transmission electron microscopy (TEM) and molecular methods and to detect the defect on tissues by the virus using histopathological method. In addition to this, sequence analysis of the isolated field strains were to be carried out and their genetic and phylogenetic closeness with the strains within the literature were to be detected. After confirming teat papillomatosis in the collected samples using histopathological and immunohistochemical methods, other diagnosis methods were then used. During the TEM examination of teat lesions, intranuclear virus particles were seen in epithelium cells. After carrying out PCR using degenerate primers and type specific primers, 7 samples were detected as positive for BPV and these samples were used for typing using sequence analysis/PCR with type-specific primers. Within these analysis, three out of seven BPV isolates we collected from infected teat tissues of different cattle were detected as BPV-6, two as BPV-10, one as BPV-2 and one as BPV-8. Five isolates we isolated during sequence analysis of positive samples were found in Xipapillomavirus 1 genus, one in Epsilonpapillomavirus 1 genus and another in Deltapapillomavirus genus. As a result, in molecular diagnosis of BPV that takes place in etiology of teat papillomas, using type specific primers proved to be useful following the usage of genotyping in molecular diagnosis of BPV and generate primers in characterization. Detecting BPV types and their prevalence, taking biosafety measures in animal breeding and giving importance to vaccine studies was considered essential.