关键词： 蝙蝠甲型流感病毒   H17N10亚型   唾液酸   RNA聚合酶  

摘要： 2012年研究人员在蝙蝠中发现了甲型流感病毒H17N10新亚型。蛋白序列及结构分析显示,HA17蛋白及NA10蛋白与已知的甲型流感病毒的血凝素和神经氨酸酶不同,它们不能与唾液酸结合,表明H17N10亚型病毒的HA17蛋白及NA10蛋白可能具有新功能。本文将扼要介绍有关蝙蝠甲型流感病毒H17N10亚型的研究进展。

关键词： emerging infectious diseases of bat origin   Hendra virus in flying-foxes   Nipah virus   severe acute respiratory syndrome coronavirus   Ebola virus   Marburg virus  

摘要： Viruses that originate in bats may be the most notorious emerging zoonoses that spill over from wildlife into domestic animals and humans. Understanding how these infections filter through ecological systems to cause disease in humans is of profound importance to public health. Transmission of viruses from bats to humans requires a hierarchy of enabling conditions that connect the distribution of reservoir hosts, viral infection within these hosts, and exposure and susceptibility of recipient hosts. For many emerging bat viruses, spillover also requires viral shedding from bats, and survival of the virus in the environment. Focusing on Hendra virus, but also addressing Nipah virus, Ebola virus, Marburg virus and coronaviruses, we delineate this cross-species spillover dynamic from the within-host processes that drive virus excretion to land-use changes that increase interaction among species. We describe how land-use changes may affect co-occurrence and contact between bats and recipient hosts. Two hypotheses may explain temporal and spatial pulses of virus shedding in bat populations: episodic shedding from persistently infected bats or transient epidemics that occur as virus is transmitted among bat populations. Management of livestock also may affect the probability of exposure and disease. Interventions to decrease the probability of virus spillover can be implemented at multiple levels from targeting the reservoir host to managing recipient host exposure and susceptibility.

关键词： 蝙蝠   SARS样冠状病毒   核苷酸序列   β干扰素   缺失突变体   转录调控  

摘要： SARS-CoV(Severe Acute Respiratory Syndrome Coronavirus)是造成2002-2003年SARS疫情的病原，造成全球8094人感染和774人死亡。在追踪SARS-CoV溯源的过程中，本实验室在菊头蝠中发现与SARS-CoV相似的病毒，命名为SARS样冠状病毒(SARS-like CoV,SL-CoV)。近期，我们分离到一株与SARS-CoV更接近的SL-CoV WIV1。该病毒可以利用人、果子狸和中华菊头蝠的ACE2作为其功能受体，并且能感染人、猪、猴以及蝙蝠的多种细胞，进一步证实蝙蝠是SARS-CoV的自然宿主。SL-CoV WIV1与人SARS-CoV（Tor2株）的全基因组核苷酸序列同源性为95％，但在其ORF6和ORF7之间存在一个额外的未知功能的ORF，命名为ORFx。在本研究中，我们探讨了SL-CoV WIV1 ORFx的功能，取得了以下结果:\n 1.用Northern blot证实WIV1野生型病毒中有十条亚基因组mRNA，而ORFx缺失突变体病毒中只有九条，由此可以确定位于ORF6与ORF7之间的ORFx可以在WIV1中转录。RT-PCR的实验结果也验证了这一结论并且确定ORFX利用了与其他ORF相同的转录调控序列。\n 2.我们在大肠杆菌中表达了全长ORFx并制备兔多克隆抗体，用间接免疫荧光的方法证明ORFx在WIV1感染的细胞中表达。\n 3.通过反向遗传学技术，我们构建了GFP替代ORFx的突变体病毒，进一步证明ORFx在病毒感染的细胞中表达，但ORFx的缺失并不影响病毒的复制。表明ORFx为一个非必需的辅助蛋白。\n 4.利用荧光素酶双报告系统研究ORFx在调节天然免疫反应中的作用，结果发现ORFx对β干扰素有微弱的抑制作用，但能够剂量依赖性激活ds-RNA介导的NF-κB活化，还可以上调宿主细胞中IL-6、IL-8和TNF的表达，推测其可能参与炎症反应。

关键词： 蝙蝠蛾拟青霉   发酵   菌粉   免疫调节  

摘要： 蝙蝠蛾拟青霉（Paecilomyces hepiali）一种冬虫夏草的无性型，其有效成分与天然虫草类似，对其进行深入开发与研究具有重要意义。本研究采用化学计量学方法和满意度函数对本实验室选育的蝙蝠蛾拟青霉菌种SY02进行发酵条件的优化。首先，采用单因素法优化蝙蝠蛾拟青霉SY02发酵培养基中的碳、氮源和无机盐的种类和浓度，然后进行部分因子试验考察培养基组分对菌株发酵的影响，并应用中心组合设计试验进一步优化培养基配方，经神经网络结合遗传算法分析获得最佳培养基配方为(％)：葡萄糖3.35%，蛋白胨1.686%，酵母浸粉1.80%，KH2PO4·3H2O0.20%，MgSO4·7H2O0.40%，ZnCl20.002%和VB10.01%。按照一定比例组成的最佳培养基配方。在此最优培养基配方下蝙蝠蛾拟青霉的发酵期望值为0.5124，较未优化期望值提高了41.925%。 然后采用Plackett-Burman设计实验筛选蝙蝠蛾拟青霉5L罐发酵条件中对产量影响显著的因素，然后对筛选出的影响显著的因素进行单因素实验优化。通过以上研究，筛选得到影响显著的因素为搅拌转速，其最佳值为300r/min。最终得到蝙蝠蛾拟青霉的5L发酵罐发酵条件为：温度26℃，接种量7%，装液量3L，种龄3d，转速300r/min，通气量150L/h，pH值自然，发酵时间4d，在此最佳条件下，蝙蝠拟青霉各有效成分的产量分别为菌丝体干重15.09g/L、多糖1.04g/L、腺苷0.07g/L、虫草酸1.81g/L、蛋白质9.40g/L。 在此基础上对蝙蝠蛾拟青霉菌粉进行了质量标准的研究，采用性状鉴别，薄层色谱法鉴别，基因鉴别的方法对样品进行了定性鉴别，从水分测定、灰分测定、重金属检查几方面对样品进行了一般杂质检查，拟将水分含量不超过7%，总灰分含量不超过1.3%，酸不溶性灰分含量不超过0.15%列入本品质量标准正文检查项下；采用高效液相色谱法对本品腺苷含量进行测定，暂定本品腺苷含量不得少于2.53mg/g. 对SY02菌粉的初步免疫活性进行研究，受试小鼠的NK细胞杀伤活性证实本菌粉可增强机体非特异性免疫功能。其次，菌粉可以使迟发型超敏反应加剧，使淋巴细胞转化率增加，二者共同证明了菌粉可以提高机体的细胞免疫功能。蝙蝠蛾拟青霉菌粉对IL10、IL12、IL12R、IFN-α等因子水平有上调效果，蝙蝠蛾拟青霉菌粉可能具有提高机体免疫功能的效果，具体的作用机制有待进一步实验研究。急性毒性试验初步证实本菌粉无毒性，可作为保健食品原料进行开发。

关键词： 蝙蝠   乙脑病毒   血清抗体   病毒血症   实验  

摘要： 研究背景和目的流行性乙型脑炎病毒(Japanese encephalitis virus, JEV,简称乙脑病毒)属于黄病毒科黄病毒属病毒,首次于1935年在日本从患者脑组织中分离获得(Nakayama病毒株,简称NAK),故又称日本脑炎病毒。乙脑病毒为单股正链RNA病毒,大小约为11 kb,只含一个长的开放阅读框架(open reading frame, ORF),编码3个结构蛋白(C蛋白、PrM or M蛋白、E蛋白)和7个非结构蛋白(NS1、NS2a、NS2b、NS3、NS4a、NS4b和NS5),分五个基因型(GⅠ、GⅡ、 GⅢ、GIV和GV)。常用的敏感实验动物是乳鼠,脑内接种乙脑病毒后3-4天发病,一周左右死亡,脑组织内含大量感染性病毒,是分离病毒、大量制备抗原的可靠方法。乳仓鼠肾细胞(Baby hamster syrian kidney cells,BHK-21)、白纹伊蚊细胞(A mosquito cells,C6/36)及鸡胚成纤维细胞则是常用的敏感细胞,病毒在这些细胞内增殖可引起细胞圆缩、颗粒增多及细胞脱落等。乙脑病毒所致的疾病为流行性乙型脑炎(Japanese encephalitis B,JE,简称乙脑),又称日本脑炎。乙脑流行于热带及亚热带地区,经蚊媒传播,主要侵犯人及某些动物的中枢神经系统,是威胁人群特别是儿童健康的重要人兽共患传染病,其潜伏期一般在10-14天左右,在临床上主要表现为高热、剧烈头痛、恶心、呕吐及意识障碍等症状,重者可出现强直痉挛、惊厥及脑膜刺激征,甚至出现呼吸衰竭而死亡等。人被带毒蚊叮咬后,大多数呈隐性感染,只有少数人发病为脑炎,发病率一般在2/10-10/10万之间,但其病死率达20%～30%,且30%～50%的幸存者有神经系统后遗症。因此,乙脑是社会和家庭的巨大疾病负担,是重要的公共卫生问题。乙脑首先于1871年在日本被发现,随后扩展到了印度、巴基斯坦、中国、澳大利亚、新加坡等国家和地区,但其主要还是流行于亚洲东南部。我国是乙脑高发地区之一,全国除了青海以外的省、市均有乙脑病例的报道,在1957年、1966和1971年曾先后发生三次乙脑暴发流行,后两次发病人数分别高达15万和17万多,发病率达20/10万以上。20世纪80年代以后,随着对适龄儿童进行乙脑疫苗尤其是减毒活疫苗的接种,我国乙脑发病率有了较大幅度的下降,并长期维持在相对较低水平,但每年的病例数仍波动于10000例左右,而且局部地区时有暴发流行。野生鸟类和猪是乙脑病毒的主要储存宿主,猪还是扩增宿主,三带喙库蚊是主要的传播媒介,而人类是乙脑病毒的终宿主。另外,一些其他种类的脊椎动物也被证明可以自然感染乙脑病毒,其中包括蝙蝠。迄今为止,已经在蝙蝠体内检测、分离到80多种病毒,例如亨德拉病毒、埃博拉病毒、冠状病毒、尼帕尔病毒,其中也包括乙脑病毒,所以蝙蝠已被认为是多种人畜共患病病毒的重要储存宿主。中国和日本的研究均表明在蝙蝠体内能检测到乙脑病毒和/或乙脑病毒的血清抗体。2008-2009年期间,本课题组亦从广东、海南及湖南的蝙蝠体内分离出4株乙脑病毒(GD1、HN2、YY87和YY158),其中GD1和HN2的全基因组序列已发表,均属于GIII型。我们课题组通过对比发现蝙蝠来源的乙脑病毒株(GD1和HN2)和人来源的乙脑病毒株(NAK)的核苷酸和氨基酸序列具有高度相似性,但是这些乙脑病毒株对蝙蝠的感染力(如毒力、免疫性反应等)是否存在差异尚不清楚。果蝠以水果为食,体型较大,易于饲养,与人类和蚊子关系密切,且曾有研究表明果蝠可通过人工注射途径而实验感染乙脑病毒。因此,为探讨人来源及蝙蝠来源乙脑病毒株对蝙蝠的感染性是否存在差异,我们选择果蝠为实验动物,进行了本实验。研究方法1.蝙蝠的捕获与处理分别于2013年6月和2014年6月,在广东和海南省部分地区捕获果蝠,捕获蝙蝠的地点主要为废旧房屋和公园棕榈树。怀孕和哺乳期的蝙蝠放生,其余的果蝠则带回实验室动物房饲养。每天喂养新鲜的水果(香蕉、番石榴、苹果和梨子等)。2.病毒株在实验中使用了1株人来源的乙脑病毒株(NAK)和2株蝙蝠来源的乙脑病毒株(HN2和GD1)。NAK病毒株为乙脑病毒原型株,于1935年从一个日本乙脑死亡病人脑组织中分离到,HN2病毒株于2008年分离自海南地区的长翼蝠,GD1病毒株于2009年分离自广东地区的大足鼠耳蝠。3株乙脑病毒在注射果蝠之前均通过BHK-21细胞进行病毒扩增,并分别计算其扩增后病毒液的半数组织培养感染剂量(Tissue culture infective dose, TCID50)。3.蝙蝠感染乙脑病毒研究中包括了2个实验。实验一有8只果蝠,随机分成3组(NAK1, HN2-1和GD1-1组),其中NAK1组有3只果蝠(bat1,bat2和b

关键词： Infectious diseases  

摘要： Australian bat lyssavirus (ABLV) infection in humans is rare but fatal, with no proven effective therapy. ABLV infection can be prevented by administration of a post-exposure prophylaxis regimen of human rabies immunoglobulin and rabies vaccine. All Australian bats (flying foxes and microbats) should be considered to be carrying ABLV unless proven otherwise. Any bat-related injury (bite, scratch or mucosal exposure to bat saliva or neural tissue) should be notified immediately to the relevant public health unit no matter how small the injury or how long ago it occurred. Human-to-human transmission of ABLV has not been reported but is theoretically possible. Standard infection control precautions should be employed when managing patients with suspected or confirmed ABLV infection.

关键词： Astroviruses   Central Europe   Coronaviruses   Calciviruses   Bats  

摘要： Background: Bat-borne viruses pose a potential risk to human health and are the focus of increasing scientific interest. To start gaining information about bat-transmitted viruses in Hungary, we tested multiple bat species for several virus groups between 2012 and 2013. Materials and Methods: Fecal samples were collected from bats across Hungary. We performed group-specific RT-PCR screening for astro-, calici-, corona-, lyssa-, othoreo-, paramyxo-, and rotaviruses. Positive samples were selected and sequenced for further phylogenetic analyses. Results: A total of 447 fecal samples, representing 24 European bat species were tested. Novel strains of astroviruses, coronaviruses, and caliciviruses were detected and analyzed phylogenetically. Out of the 447 tested samples, 40 (9%) bats were positive for at least one virus. Bat-transmitted astroviruses (BtAstV) were detected in eight species with a 6.93% detection rate (95% confidence interval [CI] 4.854, 9.571). Coronaviruses (BtCoV) were detected in seven bat species with a detection rate of 1.79% (95% CI 0.849, 3.348), whereas novel caliciviruses (BtCalV) were detected in three bat species with a detection rate of 0.67% (95% CI 0.189, 1.780). Phylogenetic analyses revealed a great diversity among astrovirus strains, whereas the Hungarian BtCoV strains clustered together with both alpha- and betacoronavirus strains from other European countries. One of the most intriguing findings of our investigation is the discovery of novel BtCalVs in Europe. The Hungarian BtCalV did not cluster with any of the calcivirus genera identified in the family so far. Conclusions: We have successfully confirmed BtCoVs in numerous bat species. Furthermore, we have described new bat species harboring BtAstVs in Europe and found new species of CalVs. Further long-term investigations involving more species are needed in the Central European region for a better understanding on the host specificity, seasonality, phylogenetic relationships, and t

关键词： 无规定马属动物疫病区   蝙蝠   种类   分布   尼帕病毒  

摘要： [目的 ]了解广东从化无规定马属动物疫病区蝙蝠的分布及其尼帕病毒感染情况。[方法 ]采用声频调查结合粘网捕捉和手抄网捕捉方法进行蝙蝠样品的采集,并用荧光定量PCR检测方法对采集的蝙蝠咽/肛拭子进行尼帕病毒进行检测。[结果 ]发现广东从化无规定马属动物疫病区的蝙蝠集群性并不明显,种群密度较低且平均,调查记录的蝙蝠大多是属于小蝙蝠亚目的食虫蝙蝠,并未发现尼帕病毒的自然宿主―狐蝠;对采集的241只蝙蝠进行尼帕病毒检测,结果均为阴性。[结论 ]本次调查未发现蝙蝠携带尼帕病毒,但也发现本地区存在较多数量和种类的蝙蝠,为预防和控制尼帕病毒传入及可能发生的流行,有必要开展对蝙蝠的长期监测。

关键词： INFLUENZA A virus   POLYMERASES   BATS   TROPISMS in viruses   HEMAGGLUTININ  

摘要： The recent identification of highly divergent influenza A viruses in bats revealed a new, geographically dispersed viral reservoir. To investigate the molecular mechanisms of host-restricted viral tropism and the potential for transmission of viruses between humans and bats, we exposed a panel of cell lines from bats of diverse species to a prototypical human-origin influenza A virus. All of the tested bat cell lines were susceptible to influenza A virus infection. Experimental evolution of human and avian-like viruses in bat cells resulted in efficient replication and created highly cytopathic variants. Deep sequencing of adapted human influenza A virus revealed a mutation in the PA polymerase subunit not previously described, M285K. Recombinant virus with the PA M285K mutation completely phenocopied the adapted virus. Adaptation of an avian virus-like virus resulted in the canonical PB2 E627K mutation that is required for efficient replication in other mammals. None of the adaptive mutations occurred in the gene for viral hemagglutinin, a gene that frequently acquires changes to recognize host-specific variations in sialic acid receptors. We showed that human influenza A virus uses canonical sialic acid receptors to infect bat cells, even though bat influenza A viruses do not appear to use these receptors for virus entry. Our results demonstrate that bats are unique hosts that select for both a novel mutation and a well-known adaptive mutation in the viral polymerase to support replication. IMPORTANCE Bats constitute well-known reservoirs for viruses that may be transferred into human populations, sometimes with fatal consequences. Influenza A viruses have recently been identified in bats, dramatically expanding the known host range of this virus. Here we investigated the replication of human influenza A virus in bat cell lines and the barriers that the virus faces in this new host. Human influenza A and B viruses infected cells from geographically and evolutionar

关键词： 云南省   病毒学   蝙蝠   疾病预防控制中心   军事医学   地方病   研究所   科学院  

摘要： 据2014年5月19日《中国科学报》报道，中国军事医学科学院军事兽医研究所涂长春联合云南省地方病防治所和成都军区疾病预防控制中心，在云南省蝙蝠病毒研究方面取得重要进展。相关成果发表于《病毒学杂志》。