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Isolation Identification & Molecular Based Investigation Of Bovine Rotavirus

By: Ambreen Masood | Prof. Dr. Tahir Yaqoob.
Contributor(s): Dr. Jawad Nazir | Dr. Sehrish Firyal.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Department of MicrobiologyDDC classification: 1707,T Dissertation note: Livestock is an important part of the economy of Pakistan. Calf's diarrhea due to group A bovine rotavirus causes high morbidity and mortality, which results in significant economic losses to livestock. In Pakistan overall prevalence of bovine rotavirus infection is 2.6%. As Pakistan is a developing country, survival of calves is really important to produce milk, meat and hides for propagation of livestock. The aim of current study was to isolate bovine rotavirus from faecal samples of diarrheic calves by antigen capture ELISA and molecular investigations. So, it was helpful to check the prevalence of bovine rotavirus in Lahore district. This study will be a milestone for better treatment strategies of calf diarrhea problem. It will also pave the way for better vaccine development strategies to cure the disease. A total of 100 diarrheic faecal samples of cattle and buffalo's calves less than three months of age were collected from Lahore district. Rotavirus screening was done by direct sandwich ELISA by using commercial Rotavirus detection kit (Cypress Diagnostics, Belgium). ELISA confirmed 12 samples to be positive for bovine rotavirus. Among 12 positive samples, 7 were found positive in buffalo calf and 5 in cattle calf. After RNA extraction and cDNA synthesis, the PCR was done for amplification of VP4 gene of all ELISA positive bovine rotavirus samples. But only 5 samples (3 buffalo calf samples and 2 cattle calf samples) give desired product of 880 bp of VP4 gene. After sequencing and bioinformatics analysis, phylogenetic tree was constructed. It is evident that Pakistani bovine rotavirus VP4 gene (BRV/QOL/13) has maximum identity of 98% with Indian bovine rotaviruses VP4 gene.
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Livestock is an important part of the economy of Pakistan. Calf's diarrhea due to group A bovine rotavirus causes high morbidity and mortality, which results in significant economic losses to livestock. In Pakistan overall prevalence of bovine rotavirus infection is 2.6%. As Pakistan is a developing country, survival of calves is really important to produce milk, meat and hides for propagation of livestock. The aim of current study was to isolate bovine rotavirus from faecal samples of diarrheic calves by antigen capture ELISA and molecular investigations. So, it was helpful to check the prevalence of bovine rotavirus in Lahore district.
This study will be a milestone for better treatment strategies of calf diarrhea problem. It will also pave the way for better vaccine development strategies to cure the disease. A total of 100 diarrheic faecal samples of cattle and buffalo's calves less than three months of age were collected from Lahore district. Rotavirus screening was done by direct sandwich ELISA by using commercial Rotavirus detection kit (Cypress Diagnostics, Belgium). ELISA confirmed 12 samples to be positive for bovine rotavirus. Among 12 positive samples, 7 were found positive in buffalo calf and 5 in cattle calf. After RNA extraction and cDNA synthesis, the PCR was done for amplification of VP4 gene of all ELISA positive bovine rotavirus samples. But only 5 samples (3 buffalo calf samples and 2 cattle calf samples) give desired product of 880 bp of VP4 gene. After sequencing and bioinformatics analysis, phylogenetic tree was constructed. It is evident that Pakistani bovine rotavirus VP4 gene (BRV/QOL/13) has maximum identity of 98% with Indian bovine rotaviruses VP4 gene.

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