新疆维吾尔自治区夏尔西里自然保护区全沟硬蜱复合感染蜱媒病原研究

目的 了解新疆维吾尔自治区(新疆)夏尔西里自然保护区全沟硬蜱蜱媒病原复合感染情况。方法 2012年6月至2013年6月在新疆夏尔西里自然保护区采用布旗法采集全沟硬蜱,通过巢式PCR方法对蜱体内莱姆病伯氏疏螺旋体[Borrelia(B.) burgdorferi] 5S~23S rRNA 基因间隔区、无形体(Anaplasma)16S rRNA基因、斑点热立克次体(Rickettsia)ompA基因、Q热贝氏柯克斯体Coxiella(C.)burneti的com1基因、巴贝西原虫(Babesia)的Nss-rRNA基因进行扩增。结果 204只全沟硬蜱中104只蜱病原检测阳性,阳性率为50.98%,其中伯氏疏螺旋体阳性率为34.31%(n=70),斑点热立克次体阳性率为28.92%(n=59),嗜吞噬细胞无形体阳性率为9.31%(n=19),未检出Q热贝氏柯克斯体和巴贝西原虫。χ²检验显示,不同病原感染率差异有统计学意义(χ²=38.517,P=0.000)。204只蜱中有39只存在复合感染,复合感染率为19.12%(39/204),在病原检测阳性蜱中复合感染率为37.50%(39/104)。伯氏疏螺旋体与斑点热立克次体复合感染率为16.18%(n=33);伯氏疏螺旋体与嗜吞噬细胞无形体复合感染率为4.90%(n=10),斑点热立克次体与嗜吞噬细胞无形体复合感染率为2.94%(n=6),3种病原体复合感染率为2.45%(n=5)。经χ²检验,3种病原混合感染率差异有统计学意义(χ²=41.328,P=0.000)。结论 新疆夏尔西里自然保护区全沟硬蜱存在伯氏疏螺旋体、嗜吞噬细胞无形体、斑点热立克次体的自然感染,并存在2~3种病原的复合感染。     

内蒙古部分边境口岸地区主要蜱类及蜱媒病原检测

目的 调查内蒙古主要陆地边境口岸地区蜱的种群分布、构成和自然感染病原体情况.方法 采用人工/小时布旗法和宿主体上搜法采集蜱标本,PCR法进行病原检测.结果 在被调查的策克、满都拉、满洲里3个口岸地区共采集蜱1313只,隶属于1科4属7种.草原革蜱在3个口岸均有分布,策克口岸获蜱占69.08%、蜱种多(6种)、短小扇头蜱为该口岸的优势种占74.86%.3个口岸地区共检测出5种蜱传疾病病原体,其中贝氏斯柯氏体仅在策克检出;平均感染率依次为莱姆病螺旋体15.08%、人巴贝西原虫3.35%、斑点热群立克次体1.98%、贝氏斯柯氏体1.07%、埃立克体0.99%.蜱感染莱姆病螺旋体阳性率在上述3个口岸地区均较高,分别为13.56%、22.88%、5.00%,且地区间差异有统计学意义;人巴贝两原虫、斑点热群立克次体地区间阳性率差异有统计学意义.结论 莱姆病螺旋体等5种蜱媒病原在策克、满都拉、满洲里口岸地区有不同程度自然感染.     

绥芬河公路口岸附近地区鼠类EHF IgG抗体调查报告

目的 了解绥芬河公路口岸附近EHF在鼠类中的分布情况.方法 对鼠类进行EHF IgG抗体监测.结果 检测142只、5种鼠类的血样,检出EHF IgG抗体阳性29例,阳性率为20.42%.其中黑线姬鼠的IgG抗体阳性率较高,为30.95%.结论 EHF宿主是属于多源性动物宿主,而黑线姬是主要贮存宿主和传染源,提示我们要注意防鼠、灭鼠.     

二连浩特口岸羊寄生蜱种类及蜱媒病原调查

目的调查二连浩特羊寄生蜱种类及蜱携带病原情况。方法对蜱进行形态学种类鉴定,然后提取单只蜱DNA基因组及总RNA,使用PCR技术对莱姆病螺旋体、Q热立克次体、巴贝西原虫、斑点热立克次体、无形体、森林脑炎病毒、布尼亚病毒、克里米亚-刚果出血热病毒等病原体进行检测。结果共采集羊寄生蜱439只,包括亚东璃眼蜱、森林革蜱、草原血蜱。从森林革蜱中检测到斑点热立克次体1例,检出率为0.227 8%,其他病原未检出。结论在二连浩特口岸地区蜱种中存在斑点热立克次体感染。     

集安口岸地区鼠类汉坦病毒自然感染情况调查

目的 了解集安口岸地区鼠类感染流行性出血热的情况.方法 采用夹夜法捕鼠,取鼠肺标本,采用RT - PCR方法检测汉坦病毒.结果 共捕鼠64只,隶属于1目2科4属5种,鼠类携带汉坦病毒率为6.25％,主要是黑线姬鼠和褐家鼠.结论 应进一步加强防鼠灭鼠和鼠类肾综合征出血热监测,严防肾综合征出血热的发生和流行.     

绥芬河、东宁口岸蜱种群及携带病原体的调查

目的 了解绥芬河、东宁口岸蜱的种群生态及其携带病原体情况.方法 采用人工小时布旗法选择4种不同生境进行蜱类采集;接种于豚鼠分离立克次体:接种于BSKⅡ培养基分离莱姆病螺旋体.结果 共采集蜱6 354只,隶属于1科3属4种;1983～1985年共采集成蜱3 539只,接种豚鼠获得立克次体14株;2002年共采集成蜱2 815只,平均密度为31.6只/人工小时,获得莱姆病螺旋体1株.结论 基本查清绥芬河、东宁口岸蜱的种群构成,两口岸密度较高,种群生态特征明显,显示存在急性蜱传斑点热和莱姆病疫源地.     

绥芬河公路口岸附近地区鼠类流行性出血热IgG抗体的调查报告

[目的]了解绥芬河公路口岸附近流行性出血热(EHF)在鼠类中的流行动态。[方法]对鼠类进行EHF IgG抗体监测。[结果]检测5种142只鼠类的血样,检出EHF IgG抗体阳性29只,阳性率为20.42％。其中黑线姬鼠的IgG抗体阳性率较高,为30.95％。[结论]EHF的宿主为多源性动物宿主,而黑线姬鼠是主要储存宿主和传染源,应在该区域加强防鼠、灭鼠工作。     

金华地区部分蜱媒传染病感染状况调查研究

目的为了解金华地区蜱与部分蜱媒传染病的种类、分布以及危害状况，建立适合于当地的蜱媒病预警和防制技术体系。方法采用夹线法，在金华某地区林区采集鼠形动物和体外寄生蜱，采用PCR和序列测定方法对该地区的蜱媒病原的感染情况进行了初步流行病学调查研究。结果在体外寄生的中华硬蜱中检出艾氏螺旋体Borreliaafzelii和普氏立克次体Rickettsia prowazekii，并发现这两种病原体的复合感染现象。结论金华地区有莱姆病和普氏立克次体存在，可能引起一些蜱媒传染病的发生，应引起公共卫生部门的重视。     

厦门地区部分人群莱姆病感染及蜱媒调查

目的了解厦门地区莱姆病感染及其蜱类生物媒介的感染情况。方法以间接免疫荧光抗体试验(IFA)检测人群血清莱姆病IgG抗体,以PCR法检测蜱类可能携带的病原体DNA片段。结果检测239份人血清,IgG抗体阳性5份,阳性率为2.09%,检测蜱媒499只均为阴性。结论厦门地区人群血清中检测出莱姆病螺旋体抗体,证实厦门地区存在莱姆病感染者。     

东北部分地区蜱携带几种病原立克次体的分子检测

目的为了解东北地区蜱中携带立克次体的情况。方法应用人工小时布旗法采集东北部分地区不同生境的游离蜱,采用PCR作斑点热立克次体、查菲埃立克体、人粒细胞无形体DNA检测。结果从东北地区全沟硬蜱和森林革蜱检出查菲埃立克体DNA,全沟硬蜱平均阳性率为3.29%,森林革蜱为1.88%;从全沟硬蜱检出人粒细胞无形体DNA,平均阳性率为2.98%;从嗜群血蜱检出斑点热立克次体DNA,其平均阳性率(6.67%)明显高于全沟硬蜱(1.69%)和森林革蜱(2.36%)。结论东北地区全沟硬蜱和森林革蜱可能是查菲埃立克体、人粒细胞无形体的主要携带媒介,嗜群血蜱主要携带斑点热立克次体,提示东北地区存在三种蜱媒立克次体病的自然疫源地。     

Isolation of Candidatus Rickettsia vini from Belgian Ixodes arboricola ticks and propagation in tick cell lines

Candidatus Rickettsia vini was originally detected in Ixodes arboricola ticks from Spain, and subsequently reported from several other Western Palearctic countries including Belgium. Recently, the bacterium was isolated in mammalian (Vero) cell culture from macerated male I. arboricola from Czech Republic, but there have been no reports of propagation in tick cells. Here we report isolation in a tick cell line of three strains of Ca. R. vini from I. arboricola collected from nests of great tits (Parus major) in Belgium. Internal organs of one male and two engorged female ticks were dissected aseptically, added to cultures of the Rhipicephalus microplus cell line BME/CTVM23 and incubated at 28 °C. Rickettsia-like bacteria were first seen in Giemsa-stained cytocentrifuge smears between 2 and 15 weeks later. Two of the isolates grew rapidly, destroying the tick cells within 2–4 weeks of onward passage in BME/CTVM23 cells, while the third isolate grew much more slowly, only requiring subculture at 4−5-month intervals. PCR amplification of bacterial 16S rRNA and Rickettsia gltA, sca4, ompB, ompA and 17-kDa genes revealed that all three isolates were Ca. R. vini, with 100 % identity to each other and to published Ca. R. vini sequences from other geographical locations. Transmission electron microscopy revealed typical single Rickettsia bacteria in the cytoplasm of BME/CTVM23 cells. The Ca. R. vini strain isolated from the male I. arboricola tick, designated Boshoek1, was tested for ability to grow in a panel of Ixodes ricinus, Ixodes scapularis and R. microplus cell lines and in Vero cells. The Boshoek1 strain grew rapidly, causing severe cytopathic effect, in the R. microplus line BME26, the I. ricinus line IRE11 and Vero cells, more slowly in the I. ricinus line IRE/CTVM19, possibly established a low-level infection in the I. ricinus line IRE/CTVM20, and failed to infect cells of any of four I. scapularis lines over a 12-week observation period. This study confirmed the applicability of the simple tick organ-cell line co-cultivation technique for isolation of tick-borne Rickettsia spp. using BME/CTVM23 cells.     

北京口岸地区鼠类和刺猬体外寄生虫的监测

目的了解北京口岸地区鼠类和刺猬体外寄生虫的种类和组成。方法采用夹夜法,在北京口岸捕获到小家鼠、褐家鼠、黑线姬鼠和刺猬,并从其体外捡取寄生虫。结果共采集到鼠类和刺猬体外寄生虫12种282只,经鉴定革螨6种61只,蜱类4种203只,恙螨1种13只,蚤类1种5只。其种类分别为毒厉螨、耶氏厉螨、纳氏厉螨、格氏血厉螨、驹鼯赫刺螨、家蝇巨螯螨、森林革蜱、草原革蜱、血红扇头蜱、长角血蜱、须纤恙螨和印鼠客蚤。结论除刺猬外,各鼠种染虫率和染虫指数较低。     

饵料消耗率检验口岸范围鼠类侵害程度的可行性研究

目的通过鼠类对不同饵料消耗率的比较,对口岸范围鼠类侵害程度进行研究。方法以盐城＂陆、海、空＂口岸范围内各类型单位室内外鼠类活动作为研究对象,根据食饵消耗率,分析阳性频度分布趋势,确立鼠类侵害程度指标值。结果根据鼠征阳性率范围均值、离散度及频度分布理论,考虑卫生检疫实践,提出盐城口岸范围内鼠征法侵害程度分级标准：Ⅰ级（不足为害）侵害率0～2%;Ⅱ级（轻度侵害）侵害率〉2%～10%;Ⅲ级（中度侵害）侵害率〉10%～30%;Ⅳ级（重度侵害）侵害率〉30%。结论提出用饵料消耗率区分口岸范围鼠类侵害程度分级标准,可与鼠征法同时或单独使用,适合口岸大范围监测要求。     

Detection of tick-borne pathogens in Rhipicephalus sanguineus sensu lato and dogs from different districts of Portugal

Dogs are highly exposed to pathogens transmitted by ectoparasites. The Mediterranean climate of Southern Europe, together with the presence of stray and/or neglected pets in close proximity with humans, contribute for tick expansion and stand for increased risk to infections in humans due to the zoonotic potential of many of these agents.The aim of this study was to perform a molecular survey in dogs (suspected of tick-borne disease and/or infested with ticks), as well as in ticks collected from those animals, from 12 districts of Portugal to investigate the occurrence of Rickettsia spp. and other tick-borne pathogens (Babesia, Ehrlichia, Anaplasma and Hepatozoon). Additionally, a serological survey of spotted fever group Rickettsia in Portuguese dogs was performed using an in-house immunofluorescence assay (IFA).A total of 200 whole-blood samples and 221 Rhipicephalus sanguineus s. l. ticks were collected from dogs.A total of 14 (7 %) blood samples and 10 (4.5 %) ticks yielded presumptively positive 420-bp amplicons using the Rickettsia spp. partial ompB nested PCR. Screening of the ompB-positive samples using the gltA gene showed 8 positive ticks. All Rickettsia ompB and gltA sequences had the highest identity with R. massiliae. The Rickettsia-positive dogs were further tested for other tick-borne pathogens and were found to be infected with Babesia spp. (n = 5), but not with Ehrlichia, Anaplasma or Hepatozoon.Of the 149 dog serum specimens tested in the serological assay, 103 (69 %) were positive for IgG antibodies against spotted fever group Rickettsia. Antibodies were found in dogs from all the studied districts, in 55 (53 %) of the stray and in 48 (47 %) of the owned dogs.Our study detected and characterized for the first time R. massiliae in dogs from Portugal, broadening the geographical range of this canine pathogen and adding knowledge to the impact of this disease in dogs.     

豫南地区蜱类分布及蜱媒病原体调查与分析

摘要:目的　探讨豫南地区蜱虫分布特点,蜱媒病原体类型及流行性特征。方法　采用布旗法、体表检蜱法采集不同环境中蜱虫,采用PCR方法对蜱媒病原体的感染情况进行流行病学调查研究。结果　3-10月份采集蜱虫730只,分2属3种,其中长角血蜱占80.71%,为当地优势蜱种;蜱虫分布呈季节性消长;经PCR扩增出2种蜱媒病原体的特异性DNA片段,比对显示与发热伴血小板减少综合征病毒（SFTSV）、嗜吞噬细胞无形体有高度同源性;SFTSV阳性率为4.90%;嗜吞噬细胞无形体阳性率为2.49%。结论　豫南地区蜱虫分布有生态及季节性规律,蜱中携带SFTSV、人粒细胞无形体病的病原体,是造成人发热伴血小板减少综合征、无形体病的主要原因,应加强疫区防控措施。     

Biotyping of IRE/CTVM19 tick cell line infected by tick-borne encephalitis virus

Ticks, being vectors for a variety of pathogens such as tick-borne encephalitis virus (TBEV), have developed defense mechanisms and pathways against infections, allowing them to control the virus at a level that does not hinder their fitness and development. At the present moment, only a few studies focused on interactions between ticks and TBEV on a molecular level have been published. Here, a possible application of MALDI-TOF MS as a research tool for the investigation of tick-virus interactions was shown. Mass spectrometry (MS) profiles of TBEV-infected and non-infected IRE/CTVM19 tick cell line were compared using principal component analysis. MS spectra were clustered based on the cultivation time of cells, but not their infection status. Nevertheless, the analysis of loading plots revealed different factors (peaks) being involved in the clustering of infected and non-infected cells. Out of them, nine were assigned with proteins: five and four for non-infected and infected cells, respectively. Peak with m/z 8565 was found to be of interest because it was suppressed upon TBEV infection and assigned to proteasome subunit alpha type (B7QE67).     

Identification of protective antigens by RNA interference for control of the lone star tick, Amblyomma americanum

The lone star tick, Amblyomma americanum, vectors pathogens of emerging diseases of humans and animals in the United States. Currently, measures are not available for effective control of A. americanum infestations. Development of vaccines directed against tick proteins may reduce tick infestations and the transmission of tick-borne pathogens. However, the limiting step in tick vaccine development has been the identification of tick protective antigens. Herein, we report the application of RNA interference (RNAi) for screening an A. americanum cDNA library for discovery of tick protective antigens that reduce tick survival and weights after feeding. Four cDNA clones, encoding for putative threonyl-tRNA synthetase (2C9), 60S ribosomal proteins L13a (2D10) and L13e (2B7), and interphase cytoplasm foci protein 45 (2G7), were selected for vaccine studies in cattle, along with subolesin, a tick protective protein identified previously. In vaccinated cattle, an overall efficacy (E) > 30% was obtained when considering the vaccine effect on both nymphs and adults, but only 2D10, 2G7 and subolesin affected both tick stages. The highest efficacy of control for adult ticks (E > 55%) was obtained in cattle vaccinated with recombinant 2G7 or subolesin. These collective results demonstrated the feasibility of developing vaccines for the control of lone star tick infestations. The use of RNAi for identification of tick protective antigens proved to be a rapid and cost-effective tool for discovery of candidate vaccine antigens, and this approach could likely be applied to other parasites of veterinary and medical importance.     

湖北省鼠情动态及防制对策

对湖北省部分城乡三年来的鼠情监测结果分析表明,我省城乡鼠种构成以三大家鼠(褐家鼠、黄胸鼠、小家鼠)为主,基本查清了与人类接触密切,且传播EHF的鼠类的分布规律,证实全省城乡鼠密度有两个高峰(4～5月和9～10月),分析出我省EHF和鼠密度两者的高峰相吻合,鼠密度高峰在病峰之前,为灭鼠防病提供了科学依据。通过几年来大规模城乡灭鼠活动,三年来全省密度逐年下降,证实我省灭鼠防病工作质量可靠,鼠密度和EHF发病率大幅度下降,与鼠情及人间疫情监测结果相吻合。     

宁波口岸蜱类及莱姆病病原体携带情况的调查研究

目的掌握宁波口岸蜱的种群特征和莱姆病病原携带情况。方法采用人工小时布旗法采蜱,用PCR法检测蜱类中携带的莱姆病DNA片段。结果采集蜱872只,隶属于1科3属3种,分别为中华硬蜱、长角血蜱、镰形扇头蜱,其中长角血蜱占97.36%,为优势种;3-9月均可捕获蜱,3-6月为高峰期,7月下旬种群数量开始急剧减少;莱姆病螺旋体DNA片段检测均呈阴性。结论查清宁波口岸蜱的种群结构,宁波口岸尚无莱姆病自然疫源地的证据。