Neonatal ishalli buzağı dışkılarında sığır rotavirüs ve sığır koronavirüs izolasyonu ve sekans analizi

Abstract

Calf diarrhea is one of the most important diseases which causes economical lose and calf deaths in neonatal period. Etiology of neonatal calf diarrhea is mainly infectious and bacteria, viruses and parasites can be the causative agents. Studies showed that viral agents, mostly bovine rotavirus (BRV) and bovine coronavirus (BCoV), are the most common causes of calf diarrhea. In the present project proposal it is aimed to isolate BRV and BCoV in feces of calves with neonatal diarrhea which were collected in previous study and stored in -80°C. For this purpose the homogenates of BRV and BCoV RT-PCR positive samples will be centrifuged and inocula were prepared by filtration of supernatants with 0.22 μm filters. Isolation of BRV and BCoV will be performed in MA104 and Vero and/or MDBK cells, respectively. Viral isolation will be controlled by visualization of cytopathic effects in infected cells. RNA extraction and BRV and/or BCoV specific RT-PCR will be performed from the cells showed cytopathic effects to confirm viral isolation. Whole genome sequences and phylogenetic analyses of viral isolates will be carried out. Viral whole genome sequencing contributes important data to molecular epidemiology and evolutionary genomics of viruses. Unlike partial sequencing, whole genome sequencing provides knowledge on epidemiology, viral pathogenicity, immunopathogenesis, etc. Only one study in Turkey reveals BRV whole genome sequencing. The studies about BCoV used only partial sequencing and there is no whole genome sequence of BCoV in Turkey. BRV and/or BCoV isolates that are circulating in Kirikkale and surrounding provinces will be determined, BCoV whole genome sequence will be determined for the first time, and genetic distance of local isolates to the other isolates from Turkey and other countries will be revealed. The experiences and isolates gained from this study will contribute to further studies about BRV and BCoV, and effective vaccines produced with local strains.