云南省新分离蠓媒病毒分子特征及其与动物疫病关系研究

A large number of domestic animal diseases, including fever, hemorrhagic disease, occurred in Yunnan province every year, whcih caused huge losses to the domestic animal industry. The aetiology of animal disease is still unknown, while the epidemic time of animal diseases concentrated during the breeding season of culicoides, but its characteristics were different from the disease caused by bluetongue virus, a culicoides-borne arbovirus, suggesting that there are likely other culicoides-borne virus diseases besides animal diseases caused by bluetongue virus. To examine the unknown culicoides-borne viruses diseases, 36 strains of culicoides-borne viruses have been isolated from culicoides collected from prevalent area of domestic animal disease in Yunnan Province, the preliminary study on these viruses indicates that there is a novel culicoiodes-borne arboviruses and its relationship with animal diseases in these region. To further elucidate the molecular characteristics and related diseases of these novel viruses, we will definitude phylogenetic and evolutionary characterization on the basis of the genome sequence of novel culicoides-borne viruses. Importantly, the the relationship between the viruses and the local animal disease will be investigated by means of specific molecular biology and serology detection method. In addtional, systematic identification and molecular characteristics of the others virus isolated from culicoides in Yunnan were preformed to determine taxonomic status, molecular genetic characteristics and the relationship between the viruses and the local animal disease. This program fouse on phthogens of animals and the prevention and control of diseases, which are of importance to veterinary service and public health in Yunnan Province.

云南省每年发生大量家养动物疾病，包括发热、出血性疾病等，导致大量牲畜死亡，给家养动物业带来巨大损失。动物疫病病因不清，且流行时间相对集中在库蠓繁殖期，但其特点与已经发现的蓝舌病毒感染不同，提示这些地区存在其它蠓虫传播的病毒性疾病。为调查未知蠓媒病毒病，本项目组在云南省主要家养动物疫区采集的库蠓中已分离到36株病毒，初步研究结果提示云南省疫区存在1种新的蠓媒病毒并与家畜动物疫病相关。为进一步阐明新分离蠓媒病毒分子特征及其与动物疫病的关系，本申请拟对该新种病毒开展系统研究，包括全基因组序列测定与分析，建立特异性分子生物学和血清学检测方法，继而开展病毒与家养动物疫病关系研究等。此外，还将对其它尚未鉴定病毒分离株开展系统鉴定，明确分类地位和分子遗传特征及其与当地动物疫病关系。本项目对于解释云南省动物疫病病原体，阐明蠓虫传播病毒与我国动物疫病关系，保障我国畜牧业健康发展和畜产品安全具有重要意义。

云南省家养动物蠓传病毒性疾病流行比较严重，给家养动物业带来巨大损失。本项目从2013到2018年在云南及其它8个省50县共采集蠓虫37万多只，采用形态学和分子生物学方法鉴定了库蠓、铗蠓、毛蠓、阿蠓、埃蠓等53种14637只，6种重要传播媒介库蠓地理分布具有明显差异。云南省14个县采集的库蠓和蚊虫病毒组学研究结果提示库蠓可能携带大约70个科的病毒，大约75%是未知病毒；库蠓中同样也检测到一些蚊传病毒，但比蚊虫还携带有更多样的感染人或动物的病毒类型，揭示了库蠓可能是一种和蚊子同样重要的病毒传播媒介，应该在将来的病原监测中占据重要位置。从采集到蠓虫中分离物190株，已经鉴定了116株，它们分别属于6个科7个属13种病毒，其中6种病毒为库蠓首次分离。遗传进化分析结果提示1种为新种病毒，3种新血清型病毒，还发现当地分离的蓝舌病毒存在不同来源病毒基因片段的重配。小鼠致病性实验结果提示7种病毒能引起乳鼠死亡，3种能引起成年鼠和4种能引起免疫缺陷小鼠死亡。新分离Tibet环状病毒能够感染成年鼠小鼠，该病毒还能感染美丽奴绵羊并出现体温升高，但不能感染半细毛羊和山羊。此外我们还建立了3种蠓传病毒核酸和血清学检测方法，开展蠓传病毒与动物疾病关系研究，结果显示当地动物普遍存在Tibet环状病毒-2、鹿流行性出血热病毒-10和盖塔病毒等多种蠓传病毒感染。研究结果丰富了我国虫媒病毒资源，对于解释云南省动物疾病病原体，阐明蠓传病毒与我国动物疫病关系，保障我国畜牧业健康发展和畜产品安全具有重要意义。项目资助发表SCI论文4篇，中文核心论文9篇，待发表4篇， 1项农业卫生标准获得资助去制定。2人晋升助理研究员，培养硕士生2名、基层学员3名。项目投入经费50万元，支出45.37万元，各项支出基本与预算相符。剩余经费4.63万元，剩余经费计划用于本项目研究后续支出。