山东省水貂五种病毒病分子流行病学调查

发布时间：2018-06-30 04:50

本文选题：水貂犬瘟热病毒 + 水貂细小病毒　；参考：《山东农业大学》2015年硕士论文

【摘要】：水貂是一种珍贵的毛皮动物,具有较高的经济价值。山东有适宜水貂生长的气候环境和丰富的饲料资源,逐步成为全国水貂养殖大省。然而随着养殖户不断扩大养殖规模、增加养殖密度,饲养管理水平又过于粗放,导致各种水貂疾病也随之发生,给水貂养殖业的健康发展带来了巨大威胁。水貂犬瘟热(Canine Distemper,CD)是由犬瘟热病毒(Canine Distemper Virus,CDV)引起的一种急性、热性、高度接触性的传染病。幼年貂感染死亡率可高达80%~90%,成年貂感染死亡率为30%~50%,患病母貂常常出现流产、空怀和胚胎吸收,是危害我国毛皮动物养殖业的主要传染病之一。水貂病毒性肠炎是由水貂细小病毒(Mink Enteritis Virus,MEV)感染引起的一种以胃肠粘膜发炎、出血及严重腹泻为主要特征的急性、烈性、高致死性和高度接触性传染病,常于7~9月份暴发流行,冬季散发,已发病的貂场常于第二年夏、秋季节再次发病。流感病毒(Influenza Virus,IV)、伪狂犬病毒(Pseudo Rabies Virus,PRV)和圆环病毒(Porcine CircoVirus,PCV)在水貂中一直都是散在发生,没有得到足够的重视,但近几年感染率有所上升。本研究对威海、临沂、日照、潍坊、聊城、济宁、滨州、菏泽八个地区的水貂进行样品采集,其中肛门拭子185份,病死貂381只(分别采集每只水貂的肝、肺、肾、脾、肠、脑),共计566份,其中233份来自威海,86份来自潍坊,171份来自临沂,31份来自日照,14份来自济宁,11份来自菏泽,11份来自滨州,9份来自聊城。用PBS对肛门拭子进行洗脱、离心,收集上清、待检;分别用研磨器对采集的肝、肺、肾、脾、肠、脑等器官组织进行研磨,用PBS制成10%的组织悬液,离心,取上清、待检。采用PCR技术和荧光PCR技术对处理好的样品进行检测。结果显示,CDV的感染率约17%,MEV的感染率为7.95%,IV的感染率为3.53%,PRV的感染率为5.82%,PCV2的感染率为3.53%;混合感染主要出现在CDV和MEV之间,混合感染率为2.83%。实验结果还显示CDV和MEV的发病时间主要集中在夏、秋季节,尤其是7~9月份,IV主要集中在冬季发病,尤其是1~3月份。另外,本研究对分离到的8株CDV毒株的H基因进行了克隆测序、分析。结果显示,8株CDV H基因的核苷酸同源性在99.0~100%之间,其中分离株SDMK07和SDMK08的基因序列同源性为100%;8株病毒H基因与参考毒株的H基因同源性范围为90.0%~99.7%,其中这8株病毒H基因与参考毒株WF1307M的H基因同源性最高,在99.1%~99.7%之间。遗传进化分析说明,这8株CDV都来自同一大分支上,可能是起源于Hamamastu毒株的野毒株。结果表明,CDV依然是危害水貂养殖的重要疫病,且CDV易与其他病原混合感染;MEV在各地区也普遍发生;IV、PRV和PCV2多为散在发生。各病毒的发病时间和病毒流行特点基本一致。本研究对水貂五种病毒病的发病情况做了科学统计,为进一步防治提供了流行病学数据。
[Abstract]:Mink is a precious fur animal with high economic value. Shandong has a suitable climatic environment for mink growth and abundant feed resources, which has gradually become a large province of mink breeding in China. However, with the continuous expansion of breeding scale, increasing the density of breeding, raising and management level is too extensive, resulting in a variety of mink diseases also occurred, to the healthy development of mink industry has brought a huge threat. Mink distemper CD is an acute, feverish and highly contagious disease caused by Canine distemper virus (CDV). The mortality rate of infantile mink infection can be as high as 80% and 90%. The mortality rate of adult mink infection is 30%. The female mink often has miscarriage, empty pregnancy and embryo absorption, which is one of the major infectious diseases that harm the fur animal breeding industry in China. Mink viral enteritis is an acute, severe, highly lethal and highly contagious infectious disease characterized by gastrointestinal mucosal inflammation, bleeding and severe diarrhea caused by mink parvovirus (Mink Enteritis virus MEV) infection. In winter, the diseased mink farm often reappeared in the following summer and autumn. Influenza virus IV, pseudorabies virus PRV and porcine Circovirus PCV have been scattered in minks and have not received enough attention, but the infection rate has increased in recent years. In this study, samples were collected from minks from eight areas of Weihai, Linyi, Rizhao, Weifang, Liaocheng, Jining, Binzhou and Heze, of which 185 anal swabs and 381 dead minks (liver, lung, kidney, spleen, intestine, respectively) were collected from each mink. Of the 566, 233 from Weihai, 86 from Weifang, 171 from Linyi, 31 from Rizhao, 14 from Jining, 11 from Heze, 11 from Binzhou, 9 from Liaocheng. The anal swabs were eluted by PBS, centrifuged, supernatant collected and examined, liver, lung, kidney, spleen, intestine, brain and other organs and tissues were ground with grinder, and 10% tissue suspension was made by PBS. PCR and fluorescence PCR techniques were used to detect the treated samples. The results showed that the infection rate of MEV was about 7.955.The infection rate of PRV was 3.53. the infection rate of PCV2 was 5.82% and the infection rate of PCV2 was 3.53.The mixed infection rate was mainly between CDV and MEV, and the mixed infection rate was 2.83%. The results also showed that the onset time of CDV and MEV was mainly in summer and autumn, especially in July and September, especially in winter, especially in January and March. In addition, the H gene of 8 CDV strains was cloned and sequenced. The results showed that the nucleotide homology of CDV H gene was between 99.0% and 100%. The homology of H gene between SDMK07 and SDMK08 was 100 and the homology range of H gene between strain H and reference strain was 90.00.The homology of H gene between these 8 strains and reference strain WF1307M was the highest, between 99.7% and 99.7%, and the homology of H gene between these 8 strains and reference strain WF1307M was between 99.7% and 99.7%, among them, the homology of H gene between SDMK07 and SDMK08 was between 99.7% and 99.7%. Genetic and evolutionary analysis showed that all of the 8 CDV strains were from the same branch, and probably originated from the wild strain of Hamamastu strain. The results showed that CDV was still an important epidemic disease in mink culture, and it was easy to mix with other pathogens to infect MEV. The onset time of each virus and the epidemic characteristics of the virus are basically the same. In this study, the incidence of five virus diseases in mink was scientifically counted, which provided epidemiological data for further prevention and control.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.92

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