关键词： bat   European   immune   lyssavirus   T cell  

摘要： European bat lyssaviruses (EBLV) types 1 and 2 are closely related to classical rabiesvirus (RABV), and are capable of causingrabies interrestrial mammals, including humans. The authors have investigated the murine host innate immune response in the brain, salivary gland, spinal cord, and blood, following peripheral in oculation with EBLV-2. In the brain, increases in Toll-like receptor-3 (TLR 3) transcript preceded overt disease, with a range of inflammatory gene transcripts increasing during the clinical stage of infection. This included transcripts for interleukin-6 (IL-6), interferon-gamma (IFN-gamma), and CXC chemokine ligand 10 (CXCL10). In the salivary gland, there was a small but significant increase of CXCL10 gene transcript and a limited increase in 2'-5' oligoadenylate synthetase (2'-5' OAS1) transcript. In the blood, there was an increase in levels of IFN-gamma and virus-neutralizing antibodies (VNAs) were detected prior to the appearance of clinical signs. These changes were associated with severe lymphocyte in filtration observed within the spinal cord and dorsal root ganglia (DRG), which was dominated by T lymphocytes and associated with widespread inflammatory changes. The authors speculate that the increase of inflammatory cytokines and chemokines in response to EBLV-2 infection leads to a dramatic increase in T-cell in filtration and provides evidence for a robust immune response to lyssavirus infection that may not commonly occur in RABV infection.

关键词： 蝙蝠   冠状病毒   血清学   分子流行病学  

摘要： 研究背景和目的 SARS爆发后,蝙蝠作为多种人兽共患病病毒的自然宿主越来越引起人们的重视。 1.蝙蝠种类多,分布广 蝙蝠隶属翼手目(Chiroptera),是哺乳类中分布最广、数量最多的动物之一,全世界大约有17科180属,约960种,其中我国有7科29属107种。翼手目分两个亚目:大蝙蝠亚目(Megachiroptera),又称狐蝠,我国有1科5属7种;小蝙蝠亚目(Microchiroptera),即通常所说的蝙蝠,我国有6科24属100种。 除南北极外,蝙蝠在世界各地都有分布,包括在高纬度地区、荒凉的沙漠和孤立的岛屿上,甚至在撒哈拉大沙漠也有蝙蝠的活动。然而,大多数蝙蝠种类生活在热带和亚热带地区。蝙蝠物种丰富,分布广泛,寿命长,并有很强的飞行能力,其中一些蝙蝠种类还有长途迁移的习性。 蝙蝠与人类接触密切。有些蝙蝠就栖息在屋檐下,或是房屋的裂缝中;另外蝙蝠在觅食时经常会误闯入居民家中;一些旅游景点的岩洞中也居住有蝙蝠。在中医里面,菊头蝠的粪便被做成“夜明沙”用于治疗眼疾;在广东,果蝠被一些餐馆作为野味摆上餐桌;湖南一些地区则相信蝙蝠可以用来治疗癫痫。 2.蝙蝠与冠状病毒 2002～2003年爆发的SARS,在确定SARS冠状病毒(SARS-CoV)的动物源性后,香港大学微生物系和中科院的研究人员分别发表文章宣布从菊头蝠科的蝙蝠中检测出蝙蝠SARS样冠状病毒(Bat-SARS CoV),与人SARS冠状病毒的核苷酸同源性＞88%,并且从菊头蝠科的成员中检出很高的抗体阳性率。因此,研究人员将冠状病毒的自然宿主锁定在蝙蝠上,并对野生蝙蝠展开了一系列的研究,结果发现,蝙蝠所携带的冠状病毒基因多样性极其丰富,迄今为止已经从蝙蝠中发现了十几种新的冠状病毒。但是通过对冠状病毒与宿主受体结合的S蛋白基因编码区的同源性分析表明,bat-SARS CoV与SARS-CoV S基因的同源性仅为64%,因此,SARS-CoV的直接祖先还未被发现,其从感染动物宿主到感染人宿主的进化历程还没有完全搞清楚,对已发现的冠状病毒进行的系统进化分析也表明还有更多新的冠状病毒没有被发现,现在已知的可能只是自然界中的冠状病毒一个很小的子集。 鉴于蝙蝠携带冠状病毒基因型多态性丰富,以及蝙蝠的生态学特性,期望从中发现其他与SARS-CoV更为同源的冠状病毒,描绘出病毒在自然宿主中的详细进化历程,有助于预防和控制SARS的再次爆发流行。因此,我们从海南、广东和湖南省部分地区收集蝙蝠标本,利用免疫学和分子生物学检测技术对蝙蝠携带冠状病毒进行一次初步的分子流行病学调查。 研究方法 1.蝙蝠标本的收集和处理 2007年5月至8月之间,在海南、广东和湖南省部分地区共收集蝙蝠468只,进行编号后,请广州大学生命科学院蝙蝠研究专家吴毅教授进行鉴定。采集蝙蝠的地方主要有废弃的房屋,或者房屋的缝隙、屋檐,以及野外阴湿的山洞。 ①血清的采集。主要用两种方法,后肢静脉采血和心脏采血。采集的血样4℃静置5h后,3000r/min离心10min,取上清,装在EP管中,编号,4℃运送回实验室,立即检测。 ②直肠样本的处理。通过无菌操作解剖蝙蝠,取肛门上端直肠组织标本约2～3cm,保存在含有RNA1ater的冻存管中,4℃运输,回实验室后,立即检测或放入-70℃保存。 2.蝙蝠血清冠状病毒抗体的SPA-ELISA检测 ①利用葡萄球菌A蛋白(Staphylococcus protein A,SPA)能够天然地与人及某些哺乳动物的IgG分子上的Fc片段结合的特性,建立了一种新的SPA-ELISA方法,用于检测蝙蝠血清冠状病毒抗体; ②利用建立的SPA-ELISA对海南、广东和湖南部分地区收集的蝙蝠血清进行检测。 3.蝙蝠肠道冠状病毒的RT-PCR检测 ①参考文献,以及对已知的冠状病毒序列进行比对,找到冠状病毒的保守序列设计引物。引物扩增的目的基因位于RNA依赖的RNA多聚酶编码区,能够扩增已知的大部分冠状病毒,包括蝙蝠冠状病毒; ②利用RT-PCR检测蝙蝠直肠样本中的冠状病毒。 4.测序和基因分析 ①对于扩增获得的目的基因进行测序; ②测序结果利用GeneBank的BLAST软件进行在线同源性分析; ③在GeneBank下载其他冠状病毒序列,利用Clustal W进行多序列比对分析; ④用MEGA 4.0软件,采用Neighbor-Joining法,Jukes-Cantor模型做进化树分析。 5.质量控制 ①枪头、EP管、冻存管、手套等实验耗材均为一次性用品; ②逆转录过程采用的枪头、EP

关键词： EBLV   bat   lyssavirus   rabies   sheep   virus  

摘要： European bat lyssaviruses (EBLVs) have been known to cross the species barrier from their native bat host to other terrestrial mammals. In this study, we have confirmed EBLV-1 and EBLV-2 susceptibility in sheep (Ovis ammon) following intracranial and peripheral (intramuscular) inoculation. Notably, mild clinical disease was observed in those exposed to virus via the intramuscular route. Following the intramuscular challenge, 75% of the animals infected with EBLV-1 and 100% of those that were challenged with EBLV-2 developed clinical signs of rabies and then recovered during the 94-day observation period. Disease pathogenesis also varied substantially between the two viruses. Infection with EBLV-1 resulted in peracute clinical signs, which are suggestive of motor neuron involvement. Antibody induction was observed and substantial inflammatrory infiltrate in the brain. In contrast, more antigen was detected in the EBLV-2-infected sheep brains but less inflammatory infiltrate and no virus neutralising antibody was evident. The latter involved a more protracted disease that was behaviour orientated. A high infectious dose was required to establish EBLV infection under experimental conditions (>= 5.0 logs/ml) but the infectious dose in field cases remains unknown. These data confirm that sheep are susceptible to infection with EBLV but that there is variability in pathogenesis including neuroinvasiveness that varies with the route of infection. This study suggests that inter-species animal-to-animal transmission of a bat variant of rabies virus to a terrestrial mammal host may be limited, and may not always result in fatal encephalitis.

关键词： Chinese horseshoe bats   coronavirus   HKU2   SARS   novel   receptor   origin  

摘要： Apart from bat-SARS-CoV, we have identified a novel group I coronavirus, bat-CoV HKU2, in Rhinolophus sinicus (Chinese horseshoe bats). Since it has been suggested that the receptor-binding motif (RBM) of SARS-CoV may have been acquired from a group I coronavirus, we conducted a surveillance study and identified bat-SARS-CoV and bat-CoV HKU2 in 8.7% and 7.5% respectively of R. sinicus in Hong Kong and Guangdong. Complete genome sequencing of four strains of bat-CoV HKU2 revealed the smallest coronavirus genome (27164 nucleotides) and a unique spike protein evolutionarily distinct from the rest of the genome. This spike protein, sharing similar deletions with other group 2 coronaviruses in its C-terminus, also contained a 15-amino acid peptide homologous to a corresponding peptide within the RBM of spike protein of SARS-CoV, which was absent in other coronaviruses except bat-SARS-CoV. These suggest a common evolutionary origin in the spike protein of bat-CoV HKU2, bat-SARS-CoV, and SARS-CoV. (c) 2007 Elsevier Inc. All rights reserved.

关键词： SARS样冠状病毒   狂犬病毒   蝙蝠  

摘要： 目的了解广西岭景镇某山村蝙蝠携带SARS样冠状病毒及狂犬病毒的情况。方法对广西岭景镇某山村开展流行病学调查,捕捉蝙蝠,采集咽拭、肛拭及粪便标本,进行RT-PCR和细胞分离培养检测狂犬病毒(Rabies virus,RV)、SARS样冠状病毒(SARS-like Coronavirus,SARS-like-CoV)。结果本次调查收集了4只犬蝠(Cynopterus sphinx)、6只普氏蹄蝠(Hipposideros pratti,)和7只中菊头蝠(Rhinolophus affinis),在采集的标本中未检测到SARS样冠状病毒及狂犬病毒。结论未发现调查的3种蝙蝠携带SARS样冠状病毒及狂犬病毒。

关键词： BATS   CORONAVIRUSES   SARS (Disease)   RESPIRATORY diseases   RNA polymerases   VIRUSES  

摘要： Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness. in humans during 2002-2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.

关键词： Animals   Brazil   Chiroptera   Molecular Sequence Data   Nucleocapsid Proteins  

摘要： Thirty-three Brazilian bat rabies viruses (RVs) were studied by sequence analysis and were compared against sequences of bat-related RVs from other regions of the Americas. Phylogenetic analysis revealed that bat-related RVs formed several monophyletic lineages and that these were associated with bat species. Brazilian bat RVs were found to include nine major lineages, one of which grouped with RVs isolated from Lasiurus spp. from different regions of the Americas. These results suggest that there is considerable diversity among Brazilian bat RV variants and that some of these RV variants may be associated with bats from other countries.

关键词： PARAMYXOVIRUSES   VIRUSES   BATS   INFECTION   MEDICAL research   IMMUNOCHEMISTRY   BIOCHEMISTRY  

摘要： Tioman virus, a relatively new paramyxovirus, was isolated from fruit bats (Pteropus species) on Tioman Island, Malaysia, in 2001. The objective of this study was to determine the prevalence of antibodies to T. virus in island inhabitants, by use of comparative ELISA and serum neutralization assays. Of the 169 human sera analyzed, 5 (similar to 3.0%) were positive for T virus, by comparative ELISA. Of these 5 sera, 3 (1.8% of the total) had neutralizing antibodies against T virus, suggesting previous infection of this study population by this virus or a similar virus.

关键词： case reports   clinical aspects   diagnosis   disease vectors   immunohistochemistry   rabies   reverse transcriptase PCR   wild animals   zoonoses  

摘要： An adult female Daubenton''s bat (Myotis daubentonii) was found in shropshire, UK, during 12 August 2007. It was taken to a wildlife rehabilitation centre, where it died on the same day after showing signs of aggression, dehydration, anorexia and inability to fly. Both the person who found the bat and the rehabilitator were bitten by the animal, and were given appropriate postexposure prophylaxis for rabies. Immunohistochemistry and reverse trancriptase-polymerase chain reaction (RT-PCR) of the dead bat were positive for European bat lyssavirus type 2 (EBLV-2) infection. Tests were also conducted 2 pipistrelle bats (Pipistrellus pipistrellus) which were found dead on 15 August within 100 m of where the Daubenton''s bat was found. Both pipistrelle bats were negative for EBLV infection. This is the sixth case of EBLV-2 infection in a bat in the UK.

关键词： lyssavirus   Germany   eptesicus fuscus   genomic insertion  

摘要： Molecular typing has consistently identified European bat lyssaviruses from Germany as EBLV type 1a (EBLV-1a). This report confirms the presence of the closely related EBLV-1b in southern Germany, a group previously reported from The Netherlands, France and Spain. Furthermore, two of three German EBLV-1b isolates contain a 6 base pair insertion within the 3' untranslated region (UTR) of the nucleoprotein gene. This feature was shared with a third EBLV-1b isolate from a region of France adjacent to the French-German border. Further investigation revealed a two base pair insertion in a single isolate of EBLV-2 at the same genomic location. Although the length of the nucleoprotein gene 3'UTRs do vary between lyssavirus genotypes, such insertions have not been recorded within genotypes and could be the result of duplications within the nucleoprotein mRNA transcript polyadenylation signal. Crown Copyright (C) 2007 Published by Elsevier B.V. All rights reserved.