China Animal Husbandry & Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (10): 4930-4940.doi: 10.16431/j.cnki.1671-7236.2025.10.038

• Preventive Veterinary Medicine • Previous Articles    

Isolation,Identification and Genetic Characteristics Analysis of Genotype G10P[11] Bovine Rotavirus in Yunnan Province

HUANG Jie1,2, ZHU Pei2, LI Zhanhong2, WANG Jinping2, HE Yuwen2, YU Jianrong3, XU Xinming4, SHA Lidong5, CHEN Peifu1, YAO Jun2   

  1. 1. College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, China;
    2. Key Laboratory of Tropical and Subtropical Animal Viral Diseases of Yunnan Province, Yunnan Academy of Animal Husbandry and Veterinary Sciences, Kunming 650224, China;
    3. Yunnan Vocational College of Agriculture, Kunming 650212, China;
    4. Animal Disease Prevention and Control Center of Lufeng City, Chuxiong Prefecture, Yunnan Province, Lufeng 651200, China;
    5. Animal Disease Prevention and Control Center of Fumin County, Kunming City, Fumin 650400, China
  • Received:2025-02-21 Published:2025-09-30

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Abstract: 【Objective】 Bovine rotavirus (BRV) type A is an important pathogen causing severe enteritis in newborn calves.The experiment aimed to understand the genotypes and molecular characteristics of the prevalent BRV strains in large-scale cattle farms in Yunnan province,providing a theoretical basis for the prevention and control of viral diarrhea in cattle farms.【Method】 Pathogen detection was carried out on 7 fecal samples of Simmental calf diarrhea from a large-scale beef cattle farm in Yunnan by RT-PCR method.Samples positive for BRV nucleic acid were subjected to continuous passage and isolation of the virus through the MA-104 cell line.The virus proliferation was detected by indirect immunofluorescence assay (IFA),and the virus growth curve was plotted.The whole genomes of the isolated strain were further sequenced.The genotypes were analyzed using online comparison software.The genetic evolutionary trees of VP4 and VP7 genes were drawn using Mega 7.0 software, respectively,and the amino acid mutations were analyzed using GeneDoc software.【Result】 7 diarrhea samples were all positive for BRV,from which a BRV strain was isolated and named YNLL-2023,with being genotyped as G10-P[11]-I2-R2-C2-M2-A3-N2-T6-E2-H2.MA-104 cells infected with the isolated strain showed obvious cytopathic effect (CPE) for 12 h.The characteristic fragment of approximately 300 bp of the VP6 gene of the isolated strain was obtained by RT-PCR amplification.IFA showed that specific green fluorescence appeared in the cytoplasm of MA-104 cells after the isolated strain infected.The growth curve showed that the virus titer continuously increased from 6 to 54 h after inoculation,reaching a peak of 105.57 TCID50/100 μL.Genetic evolution analysis indicated that the nucleotide sequence of VP4 gene of the isolated strain was in the same branch as the RVA/Yak-tc/CHN/HB-3/2021/G10P[11] strain (accession No.:ON711387.1).The nucleotide sequence of VP7 gene was in the same branch as the RVA/Bovine-wt/CHN/JS31/2020/G10 strain (accession No.:PQ332932.1) and the RVA/Bovine-wt/CHN/SD1/2023/G10 strain (accession No.：PQ332943.1).Amino acid sequence analysis demonstrated that there were 7 amino acid mutations in VP4 between the isolate and the P[11] type sheep vaccine strain,and 2 amino acid mutations in VP7 between the isolate and the G10 type human-bovine recombinant vaccine strain in neutralizing epitope region.【Conclusion】 In this experiment,one strain of type G10P[11] BRV was successfully isolated from MA-104 cells.This strain was highly similar to several strains of bovine origin in China，but there were differences in neutralizing epitopes with vaccine strains of the same genotype.

Key words: Bovine rotavirus (BRV); genotype G10P[11]; biological properties; genetic characteristics

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