Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

Gehad Elkady; Jie Zhu; Qingjie Peng; Ming Chen; Xin Liu; Yingyu Chen; Changmin Hu; Huanchun Chen; Aizhen Guo

doi:10.1016/j.meegid.2021.104715

Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

Infect Genet Evol. 2021 Apr:89:104715. doi: 10.1016/j.meegid.2021.104715. Epub 2021 Jan 9.

Authors

Gehad Elkady¹, Jie Zhu¹, Qingjie Peng², Ming Chen¹, Xin Liu³, Yingyu Chen⁴, Changmin Hu⁵, Huanchun Chen⁴, Aizhen Guo⁶

Affiliations

¹ The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China; College of Veterinary Medicine, Cooperative Innovation Centre of Substantial Pig Production, Hubei Province China, Huazhong Agricultural University, Wuhan 430070, China.
² Wuhan Keqian Biology Co. Ltd, Wuhan 430023, China.
³ College of Veterinary Medicine, Cooperative Innovation Centre of Substantial Pig Production, Hubei Province China, Huazhong Agricultural University, Wuhan 430070, China.
⁴ The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China; College of Veterinary Medicine, Cooperative Innovation Centre of Substantial Pig Production, Hubei Province China, Huazhong Agricultural University, Wuhan 430070, China; National Animal Tuberculosis Para-Reference Laboratory (Wuhan) of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, China; Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, Huazhong Agricultural University, Wuhan 430070, China.
⁵ The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China.
⁶ The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China; College of Veterinary Medicine, Cooperative Innovation Centre of Substantial Pig Production, Hubei Province China, Huazhong Agricultural University, Wuhan 430070, China; National Animal Tuberculosis Para-Reference Laboratory (Wuhan) of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, China; Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: aizhen@mail.hzau.edu.cn.

PMID: 33434703
DOI: 10.1016/j.meegid.2021.104715

Abstract

Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.

Keywords: Bovine rotavirus; Calf diarrhea; ESI-LC-MS/MS genome constellation; Isolation; RVA; RVB.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cattle
Cell Line
China
Chromatography, Liquid
Macaca mulatta
Rotavirus / isolation & purification*
Rotavirus / metabolism
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Viral Proteins / metabolism*

Substances

Viral Proteins