云南省西双版纳州蚊虫分布特点及与虫媒病毒的关系

１９８１、１９８２、１９８３、１９８６、１９８７和１９８８年在西双版纳州景洪、勐海和勐腊县（市）捕获成年雌性蚊虫８属３４种３４５０８只。夜晚在农村畜圈及其周围采获蚊虫２１种,优势蚊种为棕头库蚊和三带喙库蚊;白天在野外竹林采获蚊虫２５种,优势蚊种为贺斑伊蚊和白蚊伊蚊。从三带喙库蚊（８株）、霜背库蚊（４株）、伪杂鳞库蚊（３株）、环带库蚊（１株）、棕头库蚊（１株）、中华按蚊（３株）、刺扰伊蚊（２株）、白     

老挝波乔会晒县和敦蓬县居民区蚊虫种类调查

目的调查老挝波乔省会晒县和敦蓬县居民区成蚊种类组成,为制定当地媒介控制措施提供依据。方法采用诱蚊灯通宵捕蚊法和电动捕蚊器法采集成蚊,采用形态学方法鉴定蚊虫种类。结果共捕获蚊虫3亚科7属38种13 537只,乙型脑炎媒介三带喙库蚊和棕头库蚊属于当地优势蚊种,分别占捕获总数的75.57%(10 230/13 537)和13.61%(1 843/13 537);疟疾媒介中华按蚊和登革热媒介白纹伊蚊分别占捕获总数的0.57%(77/13 537)和0.94%(127/13 537)。结论老挝波乔省会晒县和敦蓬县蚊虫种类丰富,乙型脑炎媒介三带喙库蚊、疟疾媒介中华按蚊和登革热媒介白纹伊蚊广泛存在,提示当地存在乙型脑炎、登革热、疟疾等重要虫媒传染病流行的风险,当地应加强对上述媒介蚊虫的监测。     

云南景东县乙型脑炎流行状况调查

目的调查云南省景东县流行性乙型脑炎的流行现状。方法媒介蚊虫调查采用诱蚊灯通宵捕蚊方法,蚊虫乙脑病毒检测采取RT-PCR方法,健康人群血清乙脑病毒抗体检测采用ELISA。结果共采集到蚊虫4属15种6653只,其中三带喙库蚊和中华按蚊属于当地的优势种群,分别占总捕获蚊数的82.07%和10.46%。RT-PCR检测三带喙库蚊和中华按蚊共16批1310只,各有1株感染乙脑病毒;ELISA检测当地健康人群血清乙脑病毒抗体阳性率为72.96%,不同年龄组人群乙脑病毒抗体水平差异有统计学意义(χ2=51.589,P0.01)。结论云南景东县蚊虫种类较多,三带喙库蚊和中华按蚊有传播乙型脑炎的作用。     

甘肃省庆阳市媒介蚊虫类携带病毒调查

目的了解甘肃省庆阳市区域媒介蚊虫类分布特点及携带病毒情况,为防控虫媒病毒相关疾病提供科学依据。方法采用电子诱蚊灯、捕虫网诱捕,并进行分类;用细胞培养法分离病毒,用血清学分子生物学方法进行病毒鉴定。结果2012年捕获蚊虫5种,4238只,其中淡色库蚊2459只,占58．0％;中华按蚊794只,占18．73％;三带喙库蚊652只,占15．38％;刺扰伊蚊215只,占5．08％;济南按蚊118只,占2．8％;捕获的三带喙库蚊中分离出1株盖塔病毒,淡色库蚊检出6株库蚊黄病毒。结论淡色库蚊是庆阳市区域的优势蚊种,蚊虫携带黄病毒和盖塔病毒等虫媒病毒,应加大对该地区蚊虫及虫媒病毒的监测、调查、研究及防治工作。     

云南省乙型脑炎几个重要流行区的传播媒介调查

目的了解云南省乙型脑炎重要流行区传播媒介种群密度和地区分布。方法现场CDC诱蚊灯诱捕成蚊;吸蚊管人工捕蚊2种方法调查。结果CDC诱蚊灯诱捕成蚊法调查了5个县,捕获蚊虫6属37种8023只,其中思茅6属34种6786只,三带喙库蚊占49.01%;麻栗坡、马关、砚山、邱北、捕获蚊虫6属21种1237只,其中三带喙库蚊占71.06%;人工捕成蚊法调查了7个地区,捕获蚊虫5属21种11301只,其中思茅翠云区调查4个乡(镇)捕获蚊虫5属21种10482只,三带喙库蚊占64.44%,其它6个地区捕获蚊虫4属16种819只,乙型脑炎主要传播媒介三带喙库蚊景谷占49.13%,孟连占46.15%,陇川占35.59%;结论调查结果显示:这些地区乙型脑炎疾病传播媒介品种较多,特别是三带喙库蚊在乙型脑炎流行区种群大密度较高,地区分布较广,大量危险因素存在。     

基于核糖体 DNA 第二内转录间隔区基因的 9 种蚊虫分子鉴定

目的 分析常见 9 种蚊虫核糖体 DNA 第二内转录间隔区( rDNA-ITS2 )基因序列特征,为蚊虫种类分 子鉴定方法探讨提供依据。 方法 在山东省济宁市采集淡色库蚊、三带喙库蚊、二带喙库蚊、白纹伊蚊、刺扰伊蚊、中 华按蚊、骚扰阿蚊、常型曼蚊和黄色柯蚊成蚊,形态学种类鉴定后,提取上述 9 种蚊虫 DNA,PCR 扩增 rDNA-ITS2 基 因并测序;在 GenBank 中进行同源性比对,采用 Bioedit 7. 0 软件及 DNAMAN 软件对测序结果进行比对分析,通过 DNASTAR、ClustalX 1. 81 和 Mega6 软件分析列特征并构建系统进化树探讨系统发生关系。 结果 9 种蚊虫的 rDNA- ITS2 长度在 343 bp 与 577 bp 之间,共有 59 个保守位点、449 个变异位点、235 个简约信息位点和 191 个单态位点,9 种蚊虫种间序列同源性为 28. 21% ~ 53. 76%;所有蚊虫的 rDNA-ITS2 基因分子鉴定与形态学鉴定吻合率 100%,库 蚊属的淡色库蚊、三带喙库蚊和二带喙库蚊聚成一类,伊纹属的白纹伊蚊和刺扰伊蚊聚成一类,不同种蚊虫为独立 分支。 结论 核糖体 DNA 第二内转录间隔( rDNA-ITS2 )基因可用于蚊虫属和种的鉴定。     

云南省洱源县乙型脑炎流行病学监测

本文作者于１９８６年５月－１９８８年２月对云南省乙脑高发行区洱源县右所镇的蚊类种群组成，季节消长，成蚊自然毒率；媒蚊密度与乙脑流行关系，猪群感染乙脑动态与人群发病的关系；乙脑流行前后人群免疫水平；乙脑病毒的分离与鉴定等项目进行了系统监测研究。结果表明：中华按蚊，纹库蚊是该地区优势蚊种，１－１１月可见蚊虫活动。从人房及畜厩捕获的中华按蚊，三带喙库蚊及麻翅库蚊中分离到乙脑病毒８株。证明中华按蚊及三带喙     

云南省洱源县流行性乙型脑炎暴发流行的病原分离与鉴定

１９９１年７￣１０月,云南省洱源县发生流行性乙型脑炎暴发流行,采用滤过性试验,免疫荧光试验,交互ＨＩ试验,交互ＣＦ试验和中和试验方法,对从该县流行期采集的病人血清分离的２株病毒和从三带喙库蚁分离到的２株病毒进行鉴定,证实４株病毒均为流行性乙型脑炎病毒,从病原学证实为流行性乙型脑炎暴发流行,同时还查明了三带喙库蚊是本次乙型脑炎流行的主要传播媒介。     

云南省通海县蚊虫2株乙型脑炎病毒的分离鉴定

目的了解云南省通海县蚊虫携带的乙型脑炎病毒(JEV)情况、分子特征及基因分型。方法 2015年7月在云南省玉溪市通海县采用诱蚊灯在调查点通宵捕捉蚊虫,蚊虫研磨后分别接种BHK-21和C6/36细胞进行病毒分离,使用黄病毒属特异引物进行初步鉴定,使用乙型脑炎病毒E基因特异引物进行RT-PCR扩增及测序,使用Megalign、Genedoc、Mega7等软件进行序列分析。结果共捕获三带喙库蚊、中华按蚊、致倦库蚊和骚扰阿蚊2 300只,其中三带喙库蚊为优势蚊种(78%)。分46批进行病毒分离,获得2株阳性分离物,接种BHK-21细胞96 h,引起细胞病变。使用黄病毒属特异引物和乙型脑炎病毒E基因特异引物扩增均为阳性。序列分析结果显示,2株新分离乙型脑炎病毒与疫苗株SA14-14-2在E基因存在12个氨基酸差异位点,在8个与乙脑病毒毒力相关的位点上存在6个差异位点。E基因遗传进化分析显示,2株新分离病毒与基因Ⅰ型乙型脑炎病毒位于同一进化分支,核苷酸和氨基酸同源性分别为91.8%～99.1%和98.0%～100%。遗传进化分析显示,其与云南2016年蚊虫中分离的乙脑病毒株JEV/mosq/YN/2016亲缘关系较近,核苷酸和氨基酸同源性分别为99.1%和99.8%。结论云南省通海县分离的2株病毒鉴定为基因Ⅰ型乙型脑炎病毒,其与云南省近年来流行的毒株遗传进化关系较近。2毒株的E基因上与毒力相关的关键位点中有2个位点发生突变,结构域Ⅲ的关键抗原未发生改变,因此SA14-14-2减毒疫苗株仍可用于该地区人群的免疫接种,以预防乙型脑炎的发生和流行。     

云南省通海县蚊虫2株乙型脑炎病毒的分离鉴定北大核心CSCD

目的了解云南省通海县蚊虫携带的乙型脑炎病毒(JEV)情况、分子特征及基因分型。方法2015年7月在云南省玉溪市通海县采用诱蚊灯在调查点通宵捕捉蚊虫,蚊虫研磨后分别接种BHK-21和C6/36细胞进行病毒分离,使用黄病毒属特异引物进行初步鉴定,使用乙型脑炎病毒E基因特异引物进行RT-PCR扩增及测序,使用Megalign、Genedoc、Mega7等软件进行序列分析。结果共捕获三带喙库蚊、中华按蚊、致倦库蚊和骚扰阿蚊2300只,其中三带喙库蚊为优势蚊种(78%)。分46批进行病毒分离,获得2株阳性分离物,接种BHK-21细胞96h,引起细胞病变。使用黄病毒属特异引物和乙型脑炎病毒E基因特异引物扩增均为阳性。序列分析结果显示,2株新分离乙型脑炎病毒与疫苗株SA14-14-2在E基因存在12个氨基酸差异位点,在8个与乙脑病毒毒力相关的位点上存在6个差异位点。E基因遗传进化分析显示,2株新分离病毒与基因Ⅰ型乙型脑炎病毒位于同一进化分支,核苷酸和氨基酸同源性分别为91.8%~99.1%和98.0%~100%。遗传进化分析显示,其与云南2016年蚊虫中分离的乙脑病毒株JEV/mosq/YN/2016亲缘关系较近,核苷酸和氨基酸同源性分别为99.1%和99.8%。结论云南省通海县分离的2株病毒鉴定为基因Ⅰ型乙型脑炎病毒,其与云南省近年来流行的毒株遗传进化关系较近。2毒株的E基因上与毒力相关的关键位点中有2个位点发生突变,结构域Ⅲ的关键抗原未发生改变,因此SA14-14-2减毒疫苗株仍可用于该地区人群的免疫接种,以预防乙型脑炎的发生和流行。     

云南省景洪市虫媒病毒调查分析

近１０多年来，先后从云南省景洪市各乡镇的当地病人、猪、蝙蝠和蚊虫体内分离出乙型脑炎病毒２２株，从白纹伊蚊中分离到登革４型病毒１株，从蝙蝠和蚊体内分离到基孔肯雅病毒５株，从患急性期血液中分离到辛德毕斯病毒１株，从发热病人血液、脑炎病人脑脊液、猪血清和牛血清中分离到新环状病毒４７株，从黄胸鼠肺脏中检查出流行性出血热病毒抗原阳性６份；人群、猪恒河猴或鼠类血清中亦检出上述６种病毒的抗体，表明景洪市存在乙     

Growth characteristics of the chimeric Japanese encephalitis virus vaccine candidate, ChimeriVax-JE (YF/JE SA14--14--2), in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes

The Japanese encephalitis (JE) virus vaccine candidate, ChimeriVax-JE, which consists of a yellow fever (YF) 17D virus backbone containing the prM and E genes from the JE vaccine strain JE SA14--14--2, exhibits restricted replication in non-human primates, producing only a low-level viremia following peripheral inoculation. Although this reduces the likelihood that hematophagous insects could become infected by feeding on a vaccinated host, it is prudent to investigate the replication kinetics of the vaccine virus in mosquito species that are known to vector the viruses from which the chimera is derived. In this study ChimeriVax-JE virus was compared to its parent viruses, as well as to wild-type JE virus, for its ability to replicate in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes. Individual mosquitoes were exposed to the viruses by oral ingestion of a virus-laden blood meal or by intrathoracic (IT) virus inoculation. ChimeriVax-JE virus did not replicate following ingestion by any of the three mosquito species. Additionally, replication was not detected after IT inoculation of ChimeriVax-JE in the primary JE virus vector, Cx. tritaeniorhynchus. ChimeriVax-JE exhibited moderate growth following IT inoculation into Ae. aegypti and Ae. albopictus, reaching titers of 3.6-5.0 log(10) PFU/mosquito. There was no change in the virus genotype associated with replication in mosquitoes. Similar results were observed in mosquitoes of all three species that were IT inoculated or had orally ingested the YF 17D vaccine virus. In contrast, all mosquitoes either IT inoculated with or orally fed wild-type and vaccine JE viruses became infected, reaching maximum titers of 5.4-7.3 log(10) PFU/mosquito. These results indicate that ChimeriVax-JE virus is restricted in its ability to infect and replicate in these mosquito vectors. The low viremia caused by ChimeriVax-JE in primates and poor infectivity for mosquitoes are safeguards against secondary spread of the vaccine virus.     

Mosquito surveys carried out on Green Island, Orchid Island, and Penghu Island, Taiwan, in 2003

Field surveys of mosquitoes were carried out on Green, Orchid, and Penghu Islands in 2003 to ascertain the status of mosquito vectors. Eighteen species of mosquitoes were collected, including three species of Anopheles, four species of Aedes, eight species of Culex, two species of Armigeres, and one species of Malaya. Seventeen previously recorded species were not collected in this study but 11 species collected had not previously been recorded. Ten newly recorded species, An. maculatus, An. takasagoensis, Ae. alcasidi, Ae. lineatopennis, Ae. vexans vexans, Ar. omissus, Cx. vishnui, Cx. halifaxii, Cx. hayashii, and Cx. neomimulus, were collected on Green Island and one previously unrecorded species, Ar. subalbatus, was collected on Orchid Island. Potential vectors An. maculatus and An. sinensis, malaria vectors in Korea and Mainland China, Ae. albopictus, a vector of dengue in Taiwan and West Nile virus in the USA, Cx. vishnui and Cx. tritaeniorhynchus, Japanese encephalitis vectors in Taiwan, Ae. vexans vexans, an eastern equine encephalitis vector in the USA, and Cx. quinquefasciatus, a vector of filariasis in Taiwan and West Nile virus in the USA, were among the mosquito species collected.     

云南省澜沧江下游地区蚊虫群落及地理生态位的研究

对云南省澜沧江下游地区7县（市）的14个乡镇蚊虫群落的优势种组成及分布、集中性指数、多样性指数、均 匀度指数和地理生态位宽度进行了分析。结果表明农村畜卷及人房周围夜间活动蚊虫优势种共7个,包括三带喙库蚊、棕头库蚊、迷走按蚊、伪杂鳞库蚊、霜背库蚊、中华按蚊和菲律按蚊;野外竹林树林白天活动蚊虫优势种6个,分别为白纹伊库蚊、圆斑伊蚊、多栉领方、骚扰阿蚊、刺扰伊蚊和伪白纹伊蚊;蚊虫多样性指数变化顺序：农村畜圈及人房周围生态为勐腊县关累镇（2.132)、景洪市大勐龙乡(2.018)、勐腊县象明乡（1.892）、景洪市城郊（1.856)、孟连县勐马5级（1.842)、景洪市勐罕乡（1.823)、思茅市城郊（1.688)、勐腊县城郊（1.659)、勐海县城郊（1.557)、思 茅市思茅港（1.527)、澜沧县雅口乡（1.841)、勐海县打洛镇（1.121)、澜沧县东郎乡（0.814）和普洱县同兴乡（0.553);野外竹林树林生态为景洪市城 郊（1.841)、勐腊县城郊（1.743)、景洪市大勐龙乡（1.278)、孟连县勐马乡（1.160)、澜沧县东朗乡（1.077)、勐海县打洛镇（1.031)和澜沧县雅口乡（0.741);各地蚊虫多样性指数较大,差异不显。夜间活动蚊虫中,三带喙库蚊、迷走按蚊、棕头库蚊和中华按蚊地理生态位宽度最大,分别为:0.786、0.773、0.755和0.738;白天活动蚊虫中,白纹伊蚊地理生态位宽度最大（0.826).     

First isolation of Japanese encephalitis from Culex quinquefasciatus in Thailand

Isolation of Japanese encephalitis (JE) virus using C6/36 cell and immunofluorescence virus antigen detection techniques was attempted from female mosquitoes collected with CDC gravid traps in Samut Songkhram Province in the central region and in Phuket Province in southern Thailand, in 2003. One thousand and eighty female mosquitoes including 6 species of the Culicidae family (Culex quinquefasciatus, Cx. gelidus, Cx. tritaeniorhynchus, Cx. whitmorei, Cx. vishnui complex, Cx. s.g. culiciomyia) (pooled by specific specimen), were processed for virus isolation. Two pools of Cx. quinquefasciatus yielded a JE virus isolation. This represents the first report of JE virus isolation from Cx. quinquefasciatus in Thailand.     

Vector competence of five common mosquito species in the People's Republic of China for Western equine encephalitis virus

Two strains of the Western equine encephalitis virus (WEEV) were first detected and isolated in China in 2001. The maintenance and transmission cycles of WEEV in China are currently not well understood, and the mosquito vectors involved in these cycles are unknown. To understand the ability of the local mosquitoes in China to transmit WEEV, the vector competence of five mosquito species, namely, Culex pipiens pallens Coquillett, Cx. p. quinquefasciatus Say, Aedes (Stegomyia) albopictus Skuse, Ae. (Stegomyia) aegypti Linnaeus, and C. tritaeniorhynchus Giles, for WEEV were evaluated. Infection rates for Cx. p. pallens, Cx. p. quinquefasciatus, Cx. tritaeniorhynchus, Ae. Albopictus, and Ae. aegypti were 46%, 60%, 80%, 37%, and 25%, respectively. Dissemination rates for the same species were 60%, 61%, 75%, 55%, and 50%, respectively. Transmission rates were 41%, 53%, 57%, and 45% for Cx. p. pallens, Cx. p. quinquefasciatus, Ae. Albopictus, and Ae. Aegypti, respectively. Infection rates were significantly different between species, but the difference between dissemination and transmission rates were nonsignificant. These results suggest that several local mosquito species in China are competent laboratory vectors for WEEV.     

Growth characteristics of ChimeriVax-DEN2 vaccine virus in Aedes aegypti and Aedes albopictus mosquitoes

The chimeric yellow fever (YF) 17D-dengue type 2 (ChimeriVax-DEN2) vaccine virus developed by Acambis, Inc. (Cambridge, MA) contains the prM and E genes of wild-type (wt) dengue 2 (DEN-2) (strain PUO-218) virus in the YF vaccine virus (strain 17D) backbone. The potential of ChimeriVax-DEN2 virus to infect and be transmitted by Aedes aegypti, the principal DEN and YF virus mosquito vector, and Aedes albopictus, a species that occurs in areas of active transmission of YF and DEN viruses, was evaluated. Mosquitoes were intrathoracically (IT) inoculated with virus or were fed a virus-laden blood meal, and the replication kinetics of ChimeriVax-DEN2 were compared with the wt DEN-2 and YF 17D vaccine viruses. Replication of YF 17D virus is attenuated in cultured Ae. albopictus C6/36 mosquito cells and in Ae. aegypti and Ae. albopictus mosquitoes. Growth of ChimeriVax-DEN2 virus similarly was restricted in C6/36 cells and in mosquitoes. ChimeriVax-DEN2 replicated in 56% of IT inoculated Ae. aegypti, and virus disseminated to head tissue in 36%, with a mean viral titer of 1.8 log10 PFU/mosquito. Of mosquitoes, 16% of Ae. aegypti and 24% of Ae. albopictus were infected 14 days after a blood meal containing ChimeriVax-DEN2, but virus did not disseminate to head tissue. In contrast, DEN-2 replicated in all IT inoculated and orally infected Ae. aegypti (mean titer 5.5 log10 PFU/mosquito), and virus disseminated to head tissue in 95%. Of Ae. albopictus, 84% were infected after a blood meal containing DEN-2 virus; dissemination occurred in 36%. Replication of ChimeriVax-DEN2 virus in mosquitoes corresponded to that of YF 17D vaccine virus, which is restricted in its ability to infect and replicate in mosquitoes. Therefore, transmission of ChimeriVax-DEN2 virus by vector mosquitoes is unlikely.     

三带喙库蚊和致倦库蚊经口感染乙型脑炎病毒疫苗SA14-14-2株的研究

目的通过实验，确定被SA14—14—2乙脑减毒活疫苗免疫的宿主动物和人在被媒介蚊虫叮咬后，是否存在感染和传播的可能。方法建立三带喙库蚊和致倦库蚊的实验室种群，用乙型脑炎SA14—14—2疫苗株病毒和乙脑野毒株经口感染两种库蚊，感染后不同时间取一定数量的蚊，研磨制成悬液，应用空斑试验方法检测感染后不同时间蚊体内的病毒存在情况和病毒滴度。结果在用6．0610gPFU／ml SA14～14—2病毒经口感染的两种库蚊中，没有检测到病毒空斑，即没发现蚊虫的感染；在用较高的6．1810gPFU／ml病毒经口感染的三带喙库蚊和致倦库蚊中，分别有一组蚊虫出现低滴度感染，空斑形成单位分别是1．24和1．11log10PFU／ml；在用野毒株经口感染的两种库蚊，共计19组蚊虫中，有14组发生感染，空斑形成单位在3．18～4．7910g10PFU／ml。结论作为乙脑主要传播媒介的三带喙库蚊和致倦库蚊对SA14—14—2疫苗株病毒的经口感染和病毒在体内的复制严重受限，当叮咬接种疫苗的人后，不具备发生乙脑病毒感染和传播的能力。     

Correlation of population indices of female Culex tritaeniorhynchus with Japanese encephalitis viral activity in Kapuk, Indonesia

Nineteen consecutive monthly light trap collections of mosquitoes were made between October 1978 and April 1980 in Kapuk, Indonesia. Kapuk is a small suburb of Jakarta where pigs are raised in close proximity to rice paddies which are breeding sites for Culex tritaeniorhynchus. Japanese encephalitis (JE) virus is believed to be endemic and has been recovered from mosquitoes and pigs in the area on several occasions. A total of 18,435 female Cx. tritaeniorhynchus were allocated to 359 pools of approximately 50 per pool. Virus isolations were attempted in both Vero and BHK-21 cells and agents producing cytopathic effect were identified in a micro-neutralization test. Nineteen strains of JE were recovered from the 359 pools of Cx. tritaeniorhynchus tested. The light trap index of female Cx. tritaeniorhynchus (X) and the relative frequency of pools positive for JE (Y) for each month of the study were plotted and correlation coefficients (r) calculated after transforming the mosquito population data logarithmically and the relative frequencies of isolation by arcsine square root. The close fit of the data (p less than 0.001) to an inverse linear model (1/y = a + b log10X) suggests a close dependence of JE viral activity on the population dynamics of Cx. tritaeniorhynchus. Three additional strains of JE were recovered from other Culex spp. at the same study site. One strain each was isolated from individual pools of Cx. gelidus, Cx. vishnui and Cx. fuscocephala. Cx. tritaeniorhynchus was more frequently infected with JE than the other species tested.