关键词： 新疆   蝙蝠   病毒宏基因组学   遗传多样性  

摘要： 新疆幅员辽阔,是我国最大的省份,同时也是多种人兽共患病的自然疫源地。作为很多人兽共患病毒的自然宿主,蝙蝠在新疆的地理分布、种类和数量等背景资料目前尚不清楚,其携带的病毒多样性数据更为欠缺。为了掌握新疆伊犁州蝙蝠携带病毒的病原本底情况,本研究在2个地点采集了122只蝙蝠样品。病毒宏基因组学分析揭示这些蝙蝠携带18个科的病毒,包括噬菌体、昆虫病毒、脊椎动物病毒等。根据病毒宏基因组学结果,挑选冠状病毒、圆环病毒、星状病毒、副肠孤病毒等进行特异性PCR检测,发现其阳性率为1.3%～10.5%,其中星状病毒具有丰富的遗传多样性,部分和猪源星状病毒具有很高的同源性,提示可能发生跨种传播;而副肠孤病毒则是首次从蝙蝠中发现。本研究首次开展了新疆伊犁州蝙蝠病毒组研究,为了解新疆伊犁州蝙蝠携带病毒的多样性和本底情况提供了重要的基础数据。

关键词： Rabies   Insectivorous bats   Pathogenesis   Replication  

摘要： Rabies is a fatal and viral zoonosis that causes acute, progressive encephalitis and remains an important concern in public health. In the last few years, there has been a change in the epidemiological profile of rabies after implementing canine rabies control in the Americas, which has led to a significant increase in both human and pet cases of rabies associated with insectivorous bats. Thus, it is important to understand the pathogenesis caused by Rabies virus (RABV) isolates from insectivorous bats. Viral growth kinetics, cell-to-cell spread and virus uptake in vitro were analyzed for RABV isolates from Eptesicus furiralis and Myotis nigricans. For pathogenesis evaluation, mice were inoculated with RABV isolates from Eptesicus furiralis and Myotis nigricans, and clinical signs were observed for 40 days. We observed that the insectivorous bat strains showed a higher replication rate, faster cell-to-cell spread and delayed virus uptake in N2a cells. Furthermore, after the first sign of a clinical infection, mice infected with Myotis nigricans and Eptesicus furiralis isolates succumbed rapidly (6 +/- 9 days) compared with RABV strains associated with other reservoirs. Our results show that the insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. In addition, this study also indicates that the differences in the biological characteristics of the RABV strains are important to their pathogenicity. An enhanced understanding of rabies pathogenesis may be important for the development of novel therapies for humans and in the implementation of rabies control strategies.

关键词： mumps virus   bat-derived viruses   zoonosis   reverse genetics   viral entry   immune evasion  

摘要： Bats harbor a plethora of viruses with an unknown zoonotic potential. In-depth functional characterization of such viruses is often hampered by a lack of virus isolates. The genome of a virus closely related to human mumps viruses (hMuV) was detected in African fruit bats, batMuV. Efforts to characterize batMuV were based on directed expression of the batMuV glycoproteins or use of recombinant chimeric hMuVs harboring batMuV glycoprotein. Although these studies provided initial insights into the functionality of batMuV glycoproteins, the host range, replication competence, immunomodulatory functions, virulence, and zoonotic potential of batMuV remained elusive. Here, we report the successful rescue of recombinant batMuV. BatMuV infects human cells, is largely resistant to the host interferon response, blocks interferon induction and TNF-alpha activation, and is neurotoxic in rats. Anti-hMuV antibodies efficiently neutralize batMuV. The striking similarities between hMuV and batMuV point at the putative zoonotic potential of batMuV.

关键词： 仔猪   蝙蝠   毒源   《自然》杂志   急性腹泻   中国学者   大量死亡   致死性  

摘要： 日前,由中国学者领衔的一个国际团队在英国《自然》杂志发表报告说,他们发现一种导致猪出现致死性腹泻的新型冠状病毒源自蝙蝠,相关研究对防控新发传染病具有重要意义。2016年底,广东多个养猪场的仔猪出现严重急性腹泻并大量死亡。

关键词： 肯科伊病毒(KKV)   蛛蝇   勐腊  

摘要： 为调查云南省蝙蝠体表寄生虫蛛蝇中病原体的携带情况。采集蝙蝠体表蛛蝇标本,用细胞培养法分离病毒,对阳性分离物进行分子生物学鉴定,用RT-PCR（Reverse transcription PCR）、TCID（50）（Tissue culture infective dose）、免疫荧光等实验对病毒的特性进行研究。2012年从励腊县的棕果蝠（Rousettus leschenaulti）体表采集蛛蝇183只,利用节肢动物线粒体细胞色素氧化酶I（COI）的通用引物用PCR方法鉴定为蛛蝇科的Eucampsipoda sundaica。从这些蛛蝇中分离到1株阳性分离物MLBC1215,能引起金黄地鼠肾细胞（Golden hamster kidney cell,BHK-21）和非洲绿猴肾细胞（African green monkey kidney cell,VeroE6）出现圆缩、脱落的细胞病变（Cytopathic effect,CPE）。用RT-PCR方法扩增序列,测序结果显示MLBC1215病毒株为三节段RNA病毒,各基因组编码区序列长度分别为S基因为975核苷酸,M基因为4 568核苷酸,L基因为6 866核苷酸,3个基因节段核苷酸序列所形成的进化树拓扑构图相似,MLBC1215病毒株的S、M和L基因均与1970年泰国分离的肯科伊病毒（Kaeng Khoi orthobunyavirus,KKV）（PSC-19）处于同一进化分支。MLBC1215株的S、M、L片段与泰国PSC-19株的核苷酸的同源性分别为97.9%、94.4%和95.3%;氨基酸的同源性分别为98.7%、96.5%和98.4%。MLBC1215株在BHK-21细胞生长到60h达到最高滴度,TCID_（50）为10^（4.8）/0.1ml,随后病毒滴度逐渐下降,至144h TCID_（50）为10^（3.6）/0.1ml。MLBC1215株对小白鼠的主要器官均能引起病理改变,从而导致小白鼠发病死亡。采集的33份人血清中有9份IgG抗体与MLBC1215株有反应,检测结果为阳性,抗体滴度为4份1：20,4份1：40,1份1：80。本次研究证实云南省腊勐县蝙蝠体表吸血昆虫蛛蝇携带布尼亚病毒目KKV。

关键词： Desmodus rotundus   Innate immunity   ISGs   OAS1a   OAS1b   ADAR1   PKR  

摘要： Bats are known to harbor many zoonotic viruses, some of which are pathogenic to other mammals while they seem to be harmless in bats. As the interferon (IFN) response represents the first line of defense against viral infections in mammals, it is hypothesized that activation of the IFN system is one of the mechanisms enabling bats to co-exist with viruses. We have previously reported induction of type I IFN in a cell line from the common vampire bat, Desmodus rotundus, upon polyinosinic:polycytidylic acid (poly(I:C)) stimulation. To deepen our knowledge on D. rotundus' IFN-I antiviral response, we molecularly characterized three interferon-stimulated genes (ISGs), OAS1, PKR and ADAR1, closely implicated in the IFN-I antiviral response, and tested their functionality in our cellular model. We first found that D. rotundus encoded two OAS1 paralogs, OAS1a and OAS1b, and that the functional domains of the four ISGs characterized were highly conserved with those of other mammals. Despite their significant transcription level in the absence of stimulation, the transcription of the four ISGs characterized was enhanced by poly(I:C). In addition, the transcription of OAS1a and OAS1b appears to be differentially regulated. These findings demonstrate an active ISG antiviral response in D. rotundus in which OAS1b may play an important role. (C) 2018 Elsevier Ltd. All rights reserved.

关键词： Bat   Hepevirus   Epidemiology  

摘要： Several recent studies have reported that various bat species harbor bat hepatitis E viruses (BatHEV) belonging to the family Hepeviridae, which also contains human hepatitis E virus (HEV). The distribution and ecology of BatHEV are not well known. Here, we collected and screened 81 bat fecal samples from nine bat species in Japan to detect BatHEV RNA by RT-PCR using HEV-specific primers, and detected three positive samples. Sequence and phylogenetic analyses indicated that these three viruses were BatHEVs belonging to genus Orthohepevirus D like other BatHEV strains reported earlier in various countries. These data support the first detection of BatHEVs in Japanese microbats, indicating their wide geographical distribution among multiple bat species.

关键词： alphavirus   antibodies   arbovirus   bats   flavivirus   Grenada  

摘要： Antibody detection against selected potentially zoonotic vector-borne alphaviruses and flaviviruses was conducted on sera from bats from all six parishes in Grenada, West Indies. Sera were tested for (i) antibodies to flaviviruses West Nile virus, St. Louis encephalitis virus, Ilheus virus, Bussuquara virus (BSQV), Rio Bravo virus and all four serotypes of dengue virus (DENV) by plaque reduction neutralization test (PRNT);(ii) antibodies to alphaviruses western equine encephalitis virus, Venezuelan equine encephalitis virus and eastern equine encephalitis virus by epitope-blocking enzyme-linked immunosorbent assay (ELISA);and (iii) antibodies to the alphavirus chikungunya (CHIKV) by PRNT. Two species of fruit bats were sampled, Artibeus jamaicensis and Artibeus lituratus, all roosting in or within 1,000m of human settlements. Fifteen (36%) of the 42 bats tested for neutralizing antibodies to CHIKV were positive. The CHIKV-seropositive bats lived in localities spanning five of the six parishes. All 43 bats tested for epitope-blocking ELISA antibody to the other alphaviruses were negative, except one positive for Venezuelan equine encephalitis virus. All 50 bats tested for neutralizing antibody to flaviviruses were negative, except one that had a BSQV PRNT80 titre of 20. The CHIKV serology results indicate that bats living close to and within human settlements were exposed to CHIKV in multiple locations. Importantly, bats for this study were trapped a year after the introduction and peak of the human CHIKV epidemic in Grenada. Thus, our data indicate that bats were exposed to CHIKV possibly during a time of marked decline in human cases.

关键词： 蝙蝠流感病毒   H9N2亚型禽流感病毒   M蛋白   免疫原性  

摘要： 为比较新发蝙蝠流感病毒H17N10亚型与禽流感病毒H9N2亚型的M蛋白(包括M1和M2蛋白)的免疫原性,分别构建两种亚型流感病毒M1、M2的原核表达载体和真核表达载体,利用前期反向遗传学技术拯救获得两种重组病毒感染MDCK细胞,并分别与相应的单抗或多抗进行免疫印迹和免疫荧光鉴定。结果显示,两种亚型病毒的M1蛋白的原核表达产物均与M1多抗反应,均不与2株M1单抗反应;拯救的两种重组病毒感染细胞后及其M1蛋白的真核表达产物均与M1的单抗5F2反应,不与单抗3G8及M1的多抗反应,两种亚型的结果一致;而两种亚型的M2蛋白通过与单抗和多抗反应,出现了不一致的结果,M2的单抗仅与蝙蝠流感病毒M2的原核表达产物反应,不与H9N2禽流感病毒M2的原核表达产物反应,M2的多抗则反之;且M2的多抗仅与H9N2禽流感M2的真核表达产物以及拯救病毒反应,均不与蝙蝠流感病毒M2的真核表达产物及拯救病毒反应。本研究证实这两种亚型流感病毒的M2蛋白之间确实存在免疫原性的差异,并且筛选到了可以区分蝙蝠流感病毒和禽流感病毒的M2抗体,为进一步研究流感病毒M蛋白相关的生物学机制提供了基础。