关键词： .specimens   isolated   serum samples   genus   Institut Pasteur   Fruit bats   MADAGASCARIENSIS   Pteropus   Tioman   Pteropodid Bats   Malagasy   Eidolon   dupreanum   serum neutralization   Nipah virus   henipaviruses   TiV   Pteropodids  

摘要： Specimens were obtained from the 3 Malagasy fruit bats, Pteropus rufus, Eidolon dupreanum, and Rousettus madagascariensis. Antibodies against Nipah, Hendra, and Tioman viruses were detected by immunoassay in 23 and by serum neutralization tests in 3 of 427 serum samples, which suggests that related viruses have circulated in Maclaaascar.

关键词： 狂犬病毒   蝙蝠   流行病学   系统发生   间接免疫荧光  

摘要： 狂犬病是由狂犬病毒（Rabies virus，RABV）引起的一种人兽共患自然疫源性疾病，死亡率几乎为100％。在发达国家，人的狂犬病已极为罕见甚至完全消除，犬已经不再是狂犬病传播的主要媒介。我国人狂犬病的发病数位居世界第二，2002年以来年度发病及死亡人数均超过2000例，是严重危害我国公共卫生安全的重要疫病。从建国初至今，我国对动物狂犬病未进行过系统的流行病学统计。因此针对狂犬病的自然宿主——蝙蝠开展RABV的病原生态学研究，对于阐明我国蝠源RABV的分布及遗传衍化关系具有重要意义，对于预警、减少或消灭狂犬病对人类的威胁，具有重要的公共卫生意义。 正确的狂犬病流行病学研究方法离不开制定合理有效的调查程序，包括样品的采集、背景资料的收集整理、研究方法的选择和研究结果的分析。为了获得中国部分地区蝙蝠狂犬病的流行资料，我们首先制定了蝙蝠狂犬病流行病学研究的样品采集方案，依据卫生部发布的人狂犬病疫情公报，将南方狂犬病高发区确定为重点调查采样的地区。最终在中国南方4省（广东省、广西省、云南省和海南省）14个地区和北方吉林省的长春共采集了4科12个蝙蝠品种的1387只外表健康蝙蝠进行了本次病原流行病学调查。至课题完成，共获得蝙蝠脑组织样品1287份，蝙蝠血清样品732份。在上述样品的采／收集时，我们均建立了严格的样品登记制度，详细记录了全部样品的明细档案资料，包括采集当地的狂犬病流行情况、RABV易感动物种群及分布、人文环境等，同时还建立了严格的档案登记制度，详细记录了与样品相关的流行病学信息，这些资料的收集整理对于后期的数据分析以及防控政策的制定都具有重要意义。 引起狂犬病的RABV是弹状病毒科（Rhabdoviridae）狂犬病毒属（Lyssavirus）的成员，作为单股负链RNA病毒，其核蛋白（N）的编码基因——N基因由于型间相对保守和复制过程中的高拷贝的特点，成为病毒分型和分子诊断的目标序列。现已采用系统发生分析的方法将已发现的RABV确定为7个基因型，此外尚有4种待分型。目前我国狂犬病毒野外分离株仅限于基因Ⅰ型，但是考虑到我国地处欧亚大陆，地理环境和多种野生动物栖息的生态环境，使我们有理由怀疑我国可能存在其他基因型的狂犬病毒，即狂犬相关病毒（Rabies-related virus，RRV）。为了快速完成批量样品／不同品质样品的RABV检测，我们参考了不同基因型狂犬病毒的N基因序列，设计了nested RT-PCR引物并建立了检测方法。灵敏度试验表明，所建立的nested RT-PCR方法可检出0.2TCID50的SRV9细胞毒；灵敏度较MIT法高出100倍。特异性试验表明，本方法可以扩增狂犬病毒的全部基因型的毒株；可供检测的样品包括犬源、牛源、猪源、羊源、蝙蝠源的组织样品，可以用于我国狂犬病毒易感动物带毒的筛查。该方法已在国内多家兽医疾病监控部门应用，反馈效果很好。 通过对所收／采集到的动物样品的病原学检测，获得蝠源RABV N基因部分序列79条，蝠源RABV G基因部分序列19条。检测结果证实，广东蝙蝠脑组织RABV感染阳性率为8.22％（60／730），广西蝙蝠脑组织阳性率为3.72％（11／296），云南蝙蝠脑组织阳性率为7.14％（2／28），海南蝙蝠脑组织阳性率为2.58％（6／233）。在对蝠源RABV序列进行系统发生分析时，我们参考了GenBank收集的我国及世界范围RABV流行株、固定毒株及RRV的基因序列。结果表明，我国蝠源RABV的79条N基因部分序列，核苷酸的最低同源性达81.7％，氨基酸的最低同源性为89.4％。多序列比较证实不同地域毒株间变异的核苷酸位点和氨基酸位点具有明显的规律性；系统发生分析表明，这些毒株归属为三个进化群：Ⅰa、Ⅰb和Ⅰc；群内地域性特征明显，且同源关系较近。结合参考序列数据，证明我国动物源RABV均为基因Ⅰ型，可以划分为5个群，除蝠源RABV独立的三个群外，其他动物源RABV尚组成2个群；世界蝠源RABV流行株地域性分群明显，群间地域性分布与大陆架结构密切相关，中国流行株在其中表现出明显的地域性与进化程度上的保守性。对蝠源RABV与动物源性RABV G基因的系统发生分析结果表明我国蝠源RABV G基因与我国和其他国家动物源性RABV G基因亲缘关系较远，这可能是我国蝙蝠鲜有传播RABV的原因。 为调查我国蝙蝠感染RABV的情况，本研究利用FITC-Protein A与FITC-Protein G混合物作为荧光二抗，建立了能够检测蝙蝠血清抗体的间接免疫荧光染色法（indirect fluorescent antibody test，iFAT），应用该方法和改良的荧光抗体病毒中和试验（modified FAVN，mF

关键词： 狂犬病毒   禽流感病毒   蝙蝠  

摘要： 目的调查了解广州及周边地区蝙蝠携带狂犬病毒和禽流感病毒的情况，分析蝙蝠与人类相关疾病的关系。方法2004年9月～2005年11月，在广州市及广州市野生动物市场收集来自广州及周边地区的分属2个亚目7个种的蝙蝠共794只：犬蝠（Cynopterus sphinx）330只、普氏蹄蝠（Hipposideros pratti）7只、棕果蝠（Rousettus leschenauh）154只、普通长翼蝠（Miniopterus schreibersi）5只、小黄蝠（Scotophilus kuhlii）52只、中菊头蝠（Rhinolophus affinis）209只及小菊头蝠（Rhinolophus pusillus）37只。收集的蝙蝠分别取得标本；咽拭713份、血清452份、脑组织742份、肺组织742份及直肠组织742份共计3391份，进行RT-PCR和细胞分离培养、检测狂犬病毒（RV）与禽流感病毒（AIV）。结果收集的蝙蝠标本均未检测、分离到狂犬病毒和禽流感病毒。结论广州及周边地区的7种蝙蝠可能未携带狂犬病毒和禽流感病毒。

关键词： rabies virus   lyssavirus   bats   population dynamics   maximum likelihood   coalescent theory   phylogenetic analysis  

摘要： Rabies virus (RABV) is endemic in terrestrial mammals throughout the world and in bats on the American continent. We performed the most extensive phylogenetic analyses of bat RABV sequences undertaken to date using a variety of genes. Our study supported previous suggestions that viral sequences are grouped according to the behaviour of the host species. However, there was more genetic and geographical diversity within each phylogenetic group than previously recognised, including evidence for new groups. Furthermore, three clades of Latin American bat RABV that were distinct from the previously identified "group IV" bat RABV clade and more closely related to North American bat RABV clades, were identified. Strikingly, phylogenetic trees for the G (glycoprotein) gene had a significantly different evolutionary history to those inferred for the N (nucleoprotein) and P (phosphoprotein) genes, and an analysis of these competing topologies revealed that it is not possible on current data to resolve whether bat RABV arose from terrestrial mammal RABV, or vice-versa. Finally, using coalescent approaches, we estimated that RABV had similar rates of population growth and nucleotide substitution (similar to 2.5-4 x 10(-4) substitutions per site, per year) in both bats and terrestrial mammals, despite underlying differences in epidemiology. (c) 2006 Elsevier B.V. All rights reserved.

关键词： papillomavirus   fruit bat   Rousettus aegyptiacus   Chiroptera   carcinoma   phylogenetic analysis  

摘要： The complete genomic DNA of a novel papillomavirus (PV) was isolated from a basosquamous carcinoma on the wing of an Egyptian fruit bat (Rousettus aegyptiacus). Initial short sequences of the E1 and L1 genes of this virus were retrieved by PCR with degenerate papillomavirus-specific primers, and the entire R. aegyptiacus papillomavirus type 1 (RaPV-1) DNA was then amplified by long template PCR, cloned and sequenced with a transposon insertion method. The RaPV-1 genome counts 7970 basepairs and contains the typical papillomavirus open reading frames (ORF) (E1, E2, E4, E6, E7, L1 and L2). Based on a concatenated alignment of the E1, E2, L1 and L2 open reading frames of RaPV-1 and 46 other human and animal papillomavirus type species, a neighbor-joining phylogenetic tree was constructed. This phylogenetic analysis shows that RaPV-1 has a close-to-root position in the papillomavirus evolutionary tree. Since RaPV-1 is only distantly related to other papillomaviruses (with maximally 50% nucleotide sequence identity across the L1 open reading frame), it cannot be assigned to one of the existing papillomavirus genera and therefore represents the first member of a novel, as yet unnamed, close-to-root papillomavirus genus. This is the first time a papillomavirus has been isolated and characterized from a member of the Chiroptera order. (c) 2006 Elsevier B.V. All rights reserved.

关键词： BATS   VIRUS diseases   RNA   TISSUES   INFECTION   EPTESICUS  

摘要： Here we describe the results of experimental infections of captive big brown bats (Eptesicus fuscus) with three newly isolated bat lyssaviruses from Eurasia (Aravan, Khujand, and Irkut viruses). Infection of E. fuscus was moderate (total, 55-75%). There was no evidence of transmission to in-contact cage mates. Incubation periods for Irkut virus infection were significantly shorter (p < 0.05) than for either Aravan or Khujand virus infections. In turn, quantification of viral RNA by TaqMan PCR suggests that the dynamics of Irkut virus infection may differ from those of Aravan/Khujand virus infection. Although infectious virus and viral RNA were detected in the brain of every rabid animal, dissemination to non-neuronal tissues was limited. Levels of viral RNA in brain of Aravan/Khujand virus-infected bats was significantly correlated with the number of other tissues positive by TaqMan PCR (p < 0.05), whereas no such relationship was observed for Irkut virus infection (where viral RNA was consistently detected in all tissues other than kidney). Infectious virus was isolated sporadically from salivary glands, and both infectious virus and viral RNA were obtained from oral swabs. The detection of viral RNA in oral swabs suggests that viral shedding in saliva occurred < 5 days before the onset of clinical disease.

关键词： Fruit bats   Ebola virus   Microbial genetics   Epidemiology   Phylogenetic analysis   Wildlife   Evolutionary rate   Genetic epidemiology  

摘要： [...]according to our phylogenetic estimate, the partial L sequence of this genetic ancestor would have looked identical to that sampled from infected humans during outbreaks in late 2001 and early 2002 (Entsiami and Mendemba, Figure 1), suggesting that this ancestor could not be much older. Under such a scenario, the most recent common ancestor could not be traced back further into the past because an extremely small, effective viral population size has caused the descendents of all but one of the previously existing viral lineages to be lost. Since no trapping study on bats was undertaken before 2001, we could not directly address this issue.

关键词： 冠状病毒   病毒流行病学   基因多样性   蝙蝠   中国   野生动物   SAILS   生态系统   种间传播   生态学  

摘要： 冠状病毒可以感染多种动物,包括禽类、牲畜和人类,目前可以分成三组。冠状病毒在不同宿主之间的种间传播形成一种复杂的生态系统,目前还没有人从事这方面的研究。SAILS的爆发,以及一些新发传染病要求我们研究冠状病毒的生态学特点,尤其是在蝙蝠和其他的野生动物中。[第一段]

关键词： 尼帕病毒   蝙蝠   感染调查   泰国   免疫法检测   病毒RNA   感染情况   抗体阳性   持续感染   IgG  

摘要： 为了了解泰国蝙蝠群中尼帕病毒(首次出现学名宜合称NV)的感染情况,本文作者用酶免疫法检测1304只蝙蝠,结果发现82只蝙蝠体内尼帕病毒IgG抗体阳性,另在蝙蝠的唾液和尿液中也发现了尼帕病毒RNA,表明尼帕病毒在泰国蝙蝠体内存在持续感染。[第一段]

关键词： disease   surveillance   European bat lyssavirus   rabies   zoonoses   conservation  

摘要： Worldwide, there are more than 1100 species of the Order Chiroptera, 45 of which are present in Europe, and 16 in the UK. Bats are reservoirs of, or can be infected by, several viral diseases, including rabies virus strains (in the Lyssavirus genus). Within this genus are bat variants that have been recorded in Europe;European bat lyssavirus 1 (EBLV-1), European bat lyssavirus 2 (EBLV-2) and, four currently unclassified isolates. Since 1977, 783 cases of EBLVs (by isolation of viral RNA) have been recorded in Europe. EBLV-1 or EBLV-2 has been identified in 12 bat species, with over 95% of EBLV-1 infections identified in Eptesicus serotinus. EBLV-2 is associated with Myotis species (Myotis daubentonii and Myotis dasycneme). A programme of passive surveillance in the United Kingdom between 1987 and 2004 tested 4871 bats for lyssaviruses. Of these, four M. daubentonii (3.5796 of submitted M. daubentonii) were positive for EBLV-2. Potential bias in the passive surveillance includes possible over-representation of synanthropic species and regional biases caused by varying bat submission numbers from different parts of the UK. In 2003, active surveillance in the UK began, and has detected an antibody prevalence level of 1-5% of EBLV-2 in M. daubentonii (n = 350), and one bat with antibodies to EBLV-1 in E. serotinus (n = 52). No cases of live lyssavirus infection or lyssavirus viral RNA have been detected through active surveillance. Further research and monitoring regarding prevalence, transmission, pathogenesis and immunity is required to ensure that integrated bat conservation continues throughout Europe, whilst enabling informed policy decision regarding both human and wildlife health issues. (c) 2006 Elsevier Ltd. All rights reserved.