农业生物技术学报2024，Vol.32Issue(11)：2649-2661,13.DOI:10.3969/j.issn.1674-7968.2024.11.018
兔出血症病毒2型浙江株鉴定及致病性研究
Identification and Pathogenicity Study of a Strain of Zhejiang Rabbit hemorrhagic disease virus type 2
摘要
Abstract
Rabbit hemorrhagic disease virus type 2(RHDV2)seriously threatens the safety of Oryctolagus cuniculus breeding and production.This study aimed to identify the pathogen of suspected RHDV2 infection in a meat rabbit farm in Zhejiang province,and clarify the pathogenic characteristics of isolated strain and its influence on the structure of rabbit intestinal flora.The liver samples of dead rabbits were collected firstly,the capsid protein VP60 gene of RHDV2 was amplified by RT-PCR,and pathogens were identified by sequencing and phylogenetic tree analysis.Animal regression test was used to clarify the infection and pathogenic characteristics of isolated strain,and 16S rDNA sequencing technology was used to analyze the structural changes of cecal microbiota between the blank control group and RHDV2 group.The results of the sequencing and homology analysis showed that the target fragment amplified by RT-PCR was RHDV2 VP60 partial sequence(GenBank No.PQ215520),which was homologous to China RHDV2 strain(GenBank No.OQ570964.1)with a consistency of 98.66％.The results of animal regression test showed that all 50-day-old rabbits died within 48 h of infection,and presented typical clinical symptoms such as mouth and nose bleeding.Staining of tissue sections showed disorders of liver tissue structure,and there were obvious flake necrosis hepatocytes in the liver parenchyma,more inflammatory cell infiltration in the alveolar wall,and large-scale pulmonary edema.The liver RNA of infected rabbits was extracted for PCR amplification and sequencing,and the results showed that it was consistent with the isolated strain from the diseased rabbit farm.The results of 16S rDNA high-throughput sequencing showed significant differences in the structure and relative abundance of intestinal flora between the RHDV2 group and healthy control group,and 13 species with significant differences were screened at the genus level,indicating that RHDV2 infection altered the original microbiome of the rabbit's intestine.This study explored the pathogenicity of RHDV2 isolates in Zhejiang province for the first time,which provides a reference for the prevention,control,diagnosis and treatment of RHDV2.关键词
兔出血症病毒2型(RHDV2)/RT-PCR/病理学观察/肠道菌群Key words
Rabbit hemorrhagic disease virus type 2(RHDV2)/RT-PCR/Pathological observation/Intestinal flora分类
农业科技引用本文复制引用
郝方姣,季权安,黄叶娥,崔雪梅,胡子喆,宋厚辉,鲍国连,刘燕..兔出血症病毒2型浙江株鉴定及致病性研究[J].农业生物技术学报,2024,32(11):2649-2661,13.基金项目
浙江省自然科学基金(LTGD24C180001) （LTGD24C180001）
浙江省重点研发项目(2021C02068-7) （2021C02068-7）
国家兔产业技术体系项目(CARS-43-C-2) （CARS-43-C-2）
