江苏省莱姆病生物媒介生态及防制研究

目的：为摸清我省莱姆病生物媒介－蜱的种群、分布、消长及其带菌情况,研究防制方法。方法：用布旗法及动物体外捕捉法收集蜱,分类鉴定,并进行季节消长调查;应用直接荧光抗体染色法查蜱中肠带菌率,ＢＳＫ培养基分离病原体。现场喷洒毒死蜱和高效氯氰菊酯,比较自理前后及处理组与对照组蜱密度变化情况;使用避蚊胺作实验室和现场驱蜱效果观察。结果：我省长江南北均有蜱存在,隶属１科３属５种;蜱密度有明显的季节性。二棘血蜱     

全沟硬蜱是我国北方地区莱姆病螺旋体的主要传播媒介

研究我国北方地区莱姆病螺旋体的生物媒介—蜱的种类、带菌状况及在传播莱姆病中的作用。用布旗法和动物诱捕法在山林地区采集蜱，分类鉴定，直接荧光抗体染色法查蜱中肠带菌率，ＢＳＫ培养基分离病原体，单克隆抗体鉴定菌株。调查蜱叮咬率与人群莱姆病感染率、患病率以及蜱的季节消长与莱姆病慢性游走性红斑（ＥＣＭ）病例发生的季节性关系。在北方１０省（市、区）共采集到１００００多只蜱，分属为２科７属１１种，以全沟硬蜱为优势种，占８０％～９５％。４种蜱的中肠携带莱姆病螺旋体，全沟硬蜱的带菌率高达２０．００％～４５．００％。从４种蜱分离到９８株莱姆病螺旋体，其中９０％以上是从全沟硬蜱分离的。全沟硬蜱叮咬率高的人群，莱姆病的感染率和患病率也高。全沟硬蜱的季节消长与ＥＣＭ发生的季节性相一致。证实我国北方地区至少有４种蜱的中肠内携带莱姆病螺旋体，莱姆病螺旋体的主要生物媒介是全沟硬蜱。     

江苏省蜱及其携带莱姆病螺旋体初步调查

目的：了解我省莱姆病主要传播媒介蜱的种类、分布、消长及其携带莱姆病螺旋体情况。方法：在山区、林区用布旗法及动物体外捕捉法收集蜱，并进行病原体分离。结果：我省长江南北均有蜱存在，隶属１科２属４种；蜱密度有明显季节性，４月初出现，５～６月为高峰，８月逐渐下降，至１０月下旬消失；蜱研磨培养２５组５０管，３个月定期镜检，未发现莱姆病螺旋体。结论：我省蜱的莱姆病传播媒介和贮存宿主作用尚不能定论     

应用圆形分布法研究菌痢发病率与家蝇带菌率的关系

本文应用圆形分布法研究了人群菌痢发病率与家蝇带菌率的季节特征以及二者的关系。结果显示，人群菌痢发病的高峰日为７月２９日，高峰期为４月１７日￣１１月９日；家蝇带菌的高峰日为７月１６日，高峰期为３月５日￣１１月２６日。家蝇带菌高峰日比菌痢发病高峰日提前２周，前者的高峰期比后者提前１个月。提示家蝇带菌率高低是影响人群菌痢流行的主要因素之一。     

中国莱姆病螺旋体主要生物媒介的调查研究

为研究中国莱姆病螺旋体的生物媒介———蜱的种类、带菌状况及在传播莱姆病中的作用 ,采用布旗法和动物诱捕法在山林地区采集蜱 ,进行分类鉴定 ,应用直接荧光抗体染色法查蜱中肠带菌率 ,BSK培养基分离病原体 ,单克隆抗体鉴定菌株。调查蜱叮咬率与人群莱姆病感染率、患病率以及蜱的季节消长与莱姆病慢性游走性红斑 (ECM )病例发生的季节性关系。在全国 2 0个省 (市、区 )共采集到170 0 0多只蜱 ,分属为 2科 8属 2 3种 ,蜱类分布的地区差异较大 ,北方林区以全沟硬蜱为优势种 ,南方则以二棘血蜱和粒形硬蜱为优势种。 9种蜱的中肠携带莱姆病螺旋体 ,其带菌率以全沟硬蜱、二棘血蜱和粒形硬蜱为高 ,分别为 2 0 .0 0 %～ 4 5.0 0 %、16.0 0 %～ 4 0 .0 0 %和 2 4 .0 0 %。从 8种蜱分离出 10 8株莱姆病螺旋体 ,其单克隆抗体反应与美国B31菌株有明显的差异 ,绝大多数中国菌株对H6831呈阴性反应。蜱叮咬率高的人群 ,莱姆病的感染率和患病率也高。全沟硬蜱的季节消长与ECM发生的季节性相一致。证实多种蜱的中肠内携带莱姆病螺旋体 ;我国北方林区莱姆病螺旋体的主要生物媒介是全沟硬蜱 ,南方林区二棘血蜱和粒形硬蜱是重要的生物媒介。     

全沟硬蜱在黑龙江小兴安岭莱姆病螺旋体传播中的作用

目的：探讨全沟硬蜱在莱姆病螺旋体传播中的作用。方法：用布旗法在林区草地采蜱，并分类鉴定；用直接荧光抗体法检查蜱类中肠带菌情况，并分离菌株。结果：全沟硬蜱为当地优势蜱种，人群发病与该蜱叮咬率密切相关；该蜱中肠带菌率为４０％，并从１４组（７０只）蜱中分离出２株莱姆病螺旋体；新分离株的单克隆抗体反应与国内分离株Ｍ７相同，而与美国菌株Ｂ３１有区别。结论：当地全沟硬蜱是莱姆病螺旋体的主要生物媒介。     

新安县莱姆病血清流行病学调查

我们于１９９１－１９９２年，采用免疫荧光技术检测了新安县２６６份血清中抗莱姆病螺旋体抗体，阳性３６例，人群感染率１３．５３％，不同年龄组差异明显；蜱中肠带菌检查阳性率２６．００％，初步认为长用血蜱和微小牛蜱为传播媒介；鼠血清中也检测出了莱姆病抗体。     

吉林省莱姆病疫源地地理分布

目的了解吉林省莱姆病的疫源地情况。方法按《莱姆病流行病学个案调查表》的内容对吉林省9个地区进行了人群、动物感染率，媒介蜱带菌率，蜱活动规律、消长时间、生存条件等调查；对吉林省莱姆病自然疫源地的分布、地理景观进行分析，对山区、半山区、平原调查的数据进行统计学处理。结果吉林省9个地区35个市县、55个乡镇的3561人群蜱叮咬率为91．00％，2292人群感染率为6．20％；马、牛、羊、狗、野鼠5种动物2499份调查，感染率为22．13％；抓捕的3570媒介蜱带菌率为35．80％。结论吉林省9个地区及旅游景点存在莱姆病疫源地，莱姆病并非属林业型疾病。     

密云地区莱姆病螺旋体宿主动物和传播媒介感染状况调查

目的了解密云地区莱姆病宿主动物和传播媒介的感染情况,以及宿主动物的分布情况和传播媒介蜱的季节消长规律,为密云地区莱姆病的防制提供科学依据。方法采用布旗法捕蜱,用倒置显微镜观察蜱的形态,对蜱进行鉴定。采用夹夜法捕鼠;用BSKII培养基对蜱和鼠标本进行病原分离。采用巢式聚合酶链反应（PCR）对蜱和鼠进行病原学检测。羊血清莱姆病抗体检测采用间接免疫荧光抗体法。结果密云地区捕获的蜱均为长角血蜱,其消长曲线具有明显的季节性,巢式PCR检测蜱带菌率为10．16％;黑线姬鼠和北社鼠是该地区优势鼠种,巢式PCR检测鼠带菌率为4．26％;当地放养山羊血清学检测阳性率为12．00％。结论在蜱和鼠中均检测到莱姆病螺旋体的特异片段,表明密云地区可能为莱姆病自然疫源地。     

中国逊克和俄罗斯波亚尔科沃边境蜱类监测分析

目的监测中国逊克和俄罗斯波亚尔科沃边境蜱的种类构成和季节消长。方法采用人工布旗小时法采集蜱。结果 2012年3-9月在逊克和波亚尔科沃采获蜱1 804只,计3属4种(森林革蜱、全沟硬蜱、嗜群血蜱、日本血蜱)。优势种群为森林革蜱,其中逊克占95.57%,波亚尔科沃占90.87%。不同月份的蜱种构成有所不同。逊克蜱的活动季节在4月上半月至9月上半月(165 d),波亚尔科沃蜱的活动季节在4月下半月至9月上半月(150 d)。逊克和波亚尔科沃的森林革蜱在整个活动季节均出现1次高峰,分别在4月下半月和5月上半月。结论本次监测为中国逊克和俄罗斯波亚尔科沃蜱的种群构成,季节消长,蜱与疾病和防治研究提供了参考依据。     

Relapsing fever-like spirochetes infecting European vector tick of Lyme disease agent

To determine whether relapsing fever-like spirochetes associated with hard ticks may infect Ixodes ricinus ticks in central Europe, we screened questing ticks for 16S rDNA similar to that of Asian and American relapsing fever-like spirochetes. We compared the prevalence of these spirochetes to that of Lyme disease spirochetes transmitted by the same vector. Relapsing fever-like spirochetes infect 3.5% of questing vector ticks in our three central European sites near the Rhein Valley. These spirochetes differ genetically from their American and Asian analogs while being relatively homogeneous in the region we sampled. The Lyme disease genospecies most commonly detected in central Europe are distributed broadly, whereas those that are less frequently found appear to be place-specific. The absence of co-infected ticks suggests that relapsing fever-like and Lyme disease spirochetes may not share hosts. Exposure risk for relapsing fever-like spirochetes is similar to that of certain Lyme disease genospecies. Although many persons may be bitten by ticks infected by relapsing fever-like spirochetes, health implications remain unknown.     

Migratory passerine birds as reservoirs of Lyme borreliosis in Europe

To define the role of birds as reservoirs and disseminators of Borrelia spirochetes, we characterized tick infestation and reservoir competence of migratory passerine birds in Sweden. A total of 1,120 immature Ixodes ricinus ticks were removed from 13,260 birds and assayed by quantitative polymerase chain reaction (PCR) for Borrelia, followed by DNA sequencing for species and genotype identification. Distributions of ticks on birds were aggregated, presumably because of varying encounters with ticks along migratory routes. Lyme borreliosis spirochetes were detected in 160 (1.4%) ticks. Borrelia garinii was the most common species in PCR-positive samples and included genotypes associated with human infections. Infestation prevalence with infected ticks was 5 times greater among ground-foraging birds than other bird species, but the 2 groups were equally competent in transmitting Borrelia. Migratory passerine birds host epidemiologically important vector ticks and Borrelia species and vary in effectiveness as reservoirs on the basis of their feeding behavior.     

中国六省蜱中莱姆病螺旋体分离与鉴定

目的 了解中国6省蜱中莱姆病螺旋体携带情况.方法 在6省各选取2个采样点捕蜱,采用病原分离培养和巢式PCR方法对蜱进行莱姆病螺旋体检测,通过基因测序确定分型.结果 6省共采集到2200余只蜱,约1000只蜱用于病原分离培养,从吉林省长白县全沟硬蜱标本中分离到13株螺旋体,从贵州省道真粒形硬蜱标本中分离到9株螺旋体.1255只蜱用于PCR检测,从6省的蜱标本中检测到莱姆病螺旋体特异片段,其中吉林省(长白县27.08%、通化县20.41%)、青海省(互助县25.06%、黄南县21.11%)和贵州省(道真县25.63%)蜱中莱姆病螺旋体阳性率较高,山西省(垣曲县4.72%、交城县3.64%)蜱中莱姆病螺旋体阳性率较低.通过序列同源性分析确定吉林、青海、甘肃和山西省蜱中莱姆病螺旋体基因型均为Borrelia garinii.贵州、湖南省的基因型均为Borrelia valaisiana.结论 6省蜱中均带有莱姆病螺旋体,且带菌率有差异;山西省蜱中存在Borrelia ganmi型莱姆病螺旋体,湖南省蜱中存在Borrelia valaisiana型莱姆病螺旋体.     

Enzootic origins for clinical manifestations of Lyme borreliosis

Both early localized and late disseminated forms of Lyme borreliosis are caused by Borrelia burgdorferi senso lato. Differentiating between the spirochetes that only cause localized skin infection from those that cause disseminated infection, and tracing the group of medically-important spirochetes to a specific vertebrate host species, are two critical issues in disease risk assessment and management. Borrelia burgdorferi senso lato isolates from Lyme borreliosis cases with distinct clinical manifestations (erythema migrans, neuroborreliosis, acrodermatitis chronica atrophicans, and Lyme arthritis) and isolates from Ixodes ricinus ticks feeding on rodents, birds and hedgehogs were typed to the genospecies level by sequencing part of the intergenic spacer region. In-depth molecular typing was performed by sequencing eight additional loci with different characteristics (plasmid-bound, regulatory, and housekeeping genes). The most abundant genospecies and genotypes in the clinical isolates were identified by using odds ratio as a measure of dominance. Borrelia afzelii was the most common genospecies in acrodermatitis patients and engorged ticks from rodents. Borrelia burgdorferi senso stricto was widespread in erythema migrans patients. Borrelia bavariensis was widespread in neuroborreliosis patients and in ticks from hedgehogs, but rare in erythema migrans patients. Borrelia garinii was the dominant genospecies in ticks feeding on birds. Spirochetes in ticks feeding on hedgehogs were overrepresented in genotypes of the plasmid gene ospC from spirochetes in erythema migrans patients. Spirochetes in ticks feeding on hedgehogs were overrepresented in genotypes of ospA from spirochetes in acrodermatitis patients. Spirochetes from ticks feeding on birds were overrepresented in genotypes of the plasmid and regulatory genes dbpA, rpoN and rpoS from spirochetes in neuroborreliosis patients. Overall, the analyses of our datasets support the existence of at least three transmission pathways from an enzootic cycle to a clinical manifestation of Lyme borreliosis. Based on the observations with these nine loci, it seems to be justified to consider the population structure of B. burgdorferi senso lato as being predominantly clonal.     

我国莱姆病螺旋体生物媒介的初步调查

自1987年以来,我们在全国开展了莱姆病调查,从黑龙江、吉林、辽宁、内蒙、河北、新疆、湖北、四川、福建和北京等省、市采集的全沟硬蜱、粒形硬蜱、壤塘硬蜱、嗜群血蜱、二棘血蜱和长角血蜱分离出71株莱姆病螺旋体,其中55株从全沟硬蜱分离。全沟硬蜱是东北和西北针阔叶林区的优势蜱种,其带菌率为20~45%,表明全沟硬蜱是我国北方莱姆病螺旋体的主要生物媒介。虽然从我国南方的某些硬蜱属和血蜱属种类分离到莱姆病螺旋体,但是南方的主要生物媒介还不清楚。从我国蜱体内分离的螺旋体在超微结构、单克隆抗体反应和外表蛋白等方面与美国菌株有某些区别。     

Vector competence studies with hard ticks and Borrelia burgdorferi sensu lato spirochetes: A review

Use of emerging technology allowing for identification of genetic material from pathogens and endosymbionts in ticks collected from humans, domestic animals, wildlife, or the environment has resulted in an avalanche of new data on tick-microorganism associations. This rapidly growing stream of new information is a tremendous resource but also presents challenges, including how detection of pathogen genetic material in ticks should best be interpreted. There is a tendency in the more recent published literature to incorrectly use the term “vector” based on detection of pathogen genetic material from tick species not experimentally confirmed to serve as vectors of the pathogen in question. To serve as a vector of a horizontally maintained pathogen, such as a Borrelia burgdorferi sensu lato (s.l.) Lyme borreliosis spirochete, the tick species in question must be capable of acquiring the pathogen while feeding in the larval or nymphal stage on an infectious host, maintaining it transstadially through the molt, and then transmitting the pathogen to a naïve host while feeding in the subsequent nymphal or adult stage. This review examines the experimental evidence for and against species of hard (ixodid) ticks from different genera to serve as vectors of B. burgdorferi s.l. spirochetes. Of the 18 Ixodes species ticks evaluated to date, 13 were experimentally confirmed as vectors of B. burgdorferi s.l. spirochetes. These studies focused primarily on the three major Lyme borreliosis agents: Borrelia burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii. In striking contrast, none of 8 tick species from other genera (1 Amblyomma species, 5 Dermacentor species, and 2 Haemaphysalis species) evaluated to date were unequivocally experimentally confirmed as vectors of B. burgdorferi s.l. spirochetes. The strength of the evidence for or against each tick species to serve as a vector of B. burgdorferi s.l. spirochetes is discussed together with key knowledge gaps and research challenges.     

一组长角血蜱中检出莱姆病螺旋体和斑点热群立克次体复合感染

目的 了解浙江省山区野生动物和蜱中莱姆病、斑点热、埃立克体病(无形体病)的感染情况.方法 采用巢式PCR对采集的鼠、蜱标本进行莱姆病伯氏疏螺旋体、斑点热群立克次体、埃立克体(无形体)特异性核酸片段检测分析.结果 从121份鼠标本和105组蜱标本中检出阳性结果 14份.鼠标本中检出伯氏疏螺旋体5S～23S rDNA间隔区片段1份和埃立克体(无形体)16SrDNA 5'端片段2份.蜱标本中检出阳性11份,包括伯氏疏螺旋体5S～23S rDNA间隔区片段3份和斑点热群立克次体外膜蛋白OmpA基因5'端片段8份.其中1组长角血蜱成虫标本为伯氏疏螺旋体和斑点热群立克次体复合感染,5S～23S rRNA基因间隔区和ompA基因片段均阳性,分别与伯氏疏螺旋体法雷氏基因型和马赛立克次体株等关系较近.结论 在同一组长角血蜱成虫中同时检出莱姆病疏螺旋体和斑点热群立克次体复合感染.