摘要
为了查明湖北省某种鸡场发病鸡的病原并了解其遗传进化特征及致病性,试验首先提取发病鸡的卵泡和脾脏组织样本核酸,分别对坦布苏病毒、鸡传染性支气管炎病毒、新城疫病毒、禽流感病毒、减蛋综合征病毒和滑液囊支原体6种病原体进行检测,然后对检出的病原体进行分离培养、效价测定、遗传进化分析、中和试验和致病性试验(产蛋鸡、鸭胚和产蛋鸭)。结果表明:病料中仅坦布苏病毒核酸检测结果为阳性,新城疫病毒、禽流感病毒、减蛋综合征病毒、鸡传染性支气管炎病毒和滑液囊支原体5种病原核酸检测结果均为阴性,确定病原为坦布苏病毒。经病毒的分离培养与RT-PCR扩增后确定分离到1株坦布苏病毒,将该毒株命名为坦布苏病毒HuB株,连续传代3次获得的病毒液效价为1×10^(7.0)TCID_(50)/mL。坦布苏病毒HuB株E基因与近年来流行的坦布苏病毒参考株E基因的序列相似性为86.5%~99.8%,其中与中国鸡源参考株(GX2021和CTLN)、中国蚊源参考株(YN2020)和泰国鸭源参考株(CU56)的亲缘关系较近。坦布苏病毒HuB株能被坦布苏病毒阳性血清完全中和。接种坦布苏病毒HuB株的产蛋鸡主要表现为采食量、产蛋量明显下降,卵泡变性、液化和出血,部分鸡只表现为卵泡萎缩、腹腔中有少量干酪样物;接种坦布苏病毒HuB株的鸭胚于攻毒后96小时内全部死亡(10/10),死亡鸭胚主要表现为体表发红、出血,羽毛发育迟缓,肝脏出血、呈斑驳样;接种坦布苏病毒HuB株的产蛋鸭主要表现为停止产蛋,卵泡出血、变性、萎缩。说明成功从该种鸡场发病鸡病料中分离到1株坦布苏病毒,该毒株可能由泰国鸭源毒株变异而来,且该毒株对鸭胚有致死性,对产蛋鸭的致病性较产蛋鸡更高。
In order to identify the pathogen of the infected chickens in a chicken farm in Hubei Province,and to understand its genetic evolution characteristics and pathogenicity,in the experiment,firstly,nucleic acids were extracted from follicular and spleen tissue samples of diseased chickens;six pathogens of Tembusu virus(TMUV),avian Infectious bronchitis virus(IBV),Newcastle disease virus(NDV),avian Influenza A virus(AIV),Egg drop syndrome virus(EDSV)and Mycoplasma synoviae(MS)were tested,respectively.Then the pathogen detected was subjected to isolation and culture,titer determination,genetic evolution analysis,neutralization test and pathogenicity test(laying hen,duck embryo and laying duck).The results showed that only the nucleic acid test result of TMUV in the diseased material was positive;the nucleic acid test results of five pathogens,namely,NDV,AIV,EDSV,IBV,MS,were all negative.TMUV was identified as the cause.After isolation and culture of the virus and RT-PCR amplification,one strain of TMUV was identified,which was named TMUV HuB strain;the viral liquid obtained from three consecutive passages was determined to have a titer of 1×10^(7.0)TCID_(50)/mL.The sequence similarity between the E gene of TMUV strain HuB and the E gene of the TMUV reference strain,which had become popular in recent years,was 86.5%-99.8%;among them,it was more closely related to the reference strains of chicken-derived(GX2021 and CTLN)from China,mosquito-derived(YN2020)from China,and duck-derived(CU56)from Thailand.TMUV HuB strain could be completely neutralized by TMUV positive serum.Laying hens inoculated with the HuB strain of TMUV mainly showed a-significant decrease in feed intake and egg production,follicular degeneration,liquefaction and hemorrhage;some chickens showed follicular atrophy and a small amount of caseous material in the abdominal cavity.All duck embryos inoculated with the HuB strain of TMUV died within 96 hours after chanllege(10/10);the dead duck embryos mainly showed redness and hemorrhage on the body surface o
作者
刘涛
何平有
范京惠
尤永君
邹立宏
LIU Tao;HE Pingyou;FAN Jinghui;YOU Yongjun;ZOU Lihong(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Ringpu(Baoding)Biological Pharmaceutical Co.,Ltd.,Baoding 071000,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2024年第16期62-67,77,117,118,共9页
Heilongjiang Animal Science And veterinary Medicine
基金
保定市科技计划项目(2172P006)。
关键词
鸡源坦布苏病毒
坦布苏病毒
病毒分离
病毒鉴定
遗传进化分析
致病性试验
chicken-derived Tembusu virus
Tembusu virus
virus isolation
virus identification
genetic evolutionary analysis
pathogenicity trest
