Isolation and Molecular Characterization of Bovine Coronavirus from Calves with Acute Gastroenteritis in Egypt
Isolation and Molecular Characterization of Bovine Coronavirus from Calves with Acute Gastroenteritis in Egypt
Saad A. Moussa1, Ahmed F. Afify1*, Suzan Salah2 and Ayman Hamed3
ABSTRACT
This study was intended for the antigenic and molecular characterization of betacoronavirus 1 (bovine coronavirus) BCoV in newborn gastroenteritis calves in the Delta Region, Menofia Province, Egypt, also for isolation and identification of the local circulating BCoV strain for further diagnosis or vaccination. In cattle, Betacoronavirus 1 bovine coronavirus (BCoV) is primarily involved in enteric infections which leads to serious complication which may be fatal in young calves up to 3 months of age. A total of 20 fecal samples were collected in the winter season of 2018 and 2019, all samples were serologically screening by antigen-capture ELISA, then molecular confirmation by RT-PCR targeted to nucleocapsid protein gene (N-gene) was carried out followed by phylogenetic analysis. Positive samples were isolated on Vero cell culture and identified by TEM and immune-peroxidase technique. Betacoronavirus 1 was detected by ELISA in 6 out of 20 fecal samples (30%), PCR detected 4 out of 6 ELISA positive samples at specific M.W. band of 236 bp by electrophoresis, and one sample was sequenced and submitted on Genbank with acc.no. MW173144, further phylogenetic sequence analysis revealed high percentage of identity with reference strains from different countries. Phylogenetic tree cleared that current research strain was found related to France strains 2013 and 2014, also to local Egypt strain 2019 with acc.no. MN053321, two samples were successfully isolated in Vero cells and positively identified after the 3rd passage by TEM and immune-peroxidase technique. ELISA results are considered an alarmingly high prevalence that requires further statistically designed epidemiological studies, phylogenetic analysis revealed minimum evolution rate and stability of BCoV genome.
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