长角血蜱和微小扇头蜱的形态与分子生物学鉴定

目的 建立鉴定长角血蜱和微小扇头蜱的形态学和分子生物学相结合的方法.方法 采集自湖北和河南省布尼亚新病毒疫区家养动物体表寄生的蜱及草丛、灌木中的蜱;先以形态学鉴定,后用PCR方法扩增得到蜱的12S rDNA,测序后进行同源性和系统进化分析.结果 形态学方法鉴定采集到两种蜱:长角血蜱(Haemaphysalis longicornis)和微小扇头蜱(Rhipicephalus microplus).蜱的12S rDNA经克隆、测序,用PAUP 4.0软件构建系统发生树,湖北、河南省两种蜱的12S rDNA序列分别与长角血蜱和微小扇头蜱聚类,与形态学鉴定结果一致.结论 在传统形态学分类的基础上结合分子生物学鉴定方法能更准确地鉴定蜱的种类.

Abstract：

Objective To establish a method combined morphology and molecular marker for identifying Haemaphysalis longicomis and Rhipicephalus microplus. Methods Ticks were collected from domestic animals and wild environment in epidemic area of Hubei and Henan provinces where cases of fever with thrombocytopenia syndrome were prevalent. We classified the ticks by morphology characteristics before 12S rDNA of ticks were amplified by PCR and subsequently sequenced. Phylogenetic tree was constructed by PAUP4.0. Results The ticks belonged to Haemaphysalis longicomis and Rhipicephalus microplus through observation and analysed by the morphological characteristics of the ticks. 12S rDNA was cloned and sequenced while data confirmed the morphological identification of the results. Conclusion The method based on morphology that combined with molecular marker seemed a good method for the identificaton of ticks.     

Human Rickettsia heilongjiangensis Infection, Japan

A case of Rickettsia heilongjiangensis infection in Japan was identified in a 35-year-old man who had rash, fever, and eschars. Serum contained R. heilongjiangensis antibodies, and eschars contained R. heilongjiangensis DNA. R. heilongjiangensis was also isolated from ticks in the suspected geographic area of infection.     

河北省西部山区蜱传斑点热群立克次体分子流行病学研究

目的 调查河北省西部山区蜱类斑点热群立克次体带菌情况,为蜱传斑点热群防控提供科学依据.方法 根据已发表斑点热群立克次体OmpA外膜蛋白基因序列设计特异性引物,对河北省西部山区采集的长角血蜱进行PCR检测,并对阳性样本进行测序和序列分析,建立分子系统进化树.结果 在1227份蜱DNA样本中检测出91份阳性,阳性率为7.42％;序列分析结果显示河北省西部山区长角血蜱携带立克次体与河北株暂定种(登录号:HQ651815、HQ651817、HQ651818、HQ651819、HQ651823、HQ651824)、福建株FUJ(登录号:AF169629)同处于一个分支,与河北株立克次体暂定种同源性最高(99.02％),其次是福建株(98.50％)、黑龙江立克次体绥芬株HLJ-054(登录号:AF179362)和黑龙江立克次体虎林株HL-93(登录号:AF179364)(98.13％)、日本株YM(登录号:U43795)(97.92％).结论 河北省西部山区蜱传斑点热群立克次体感染较为严重,应尽快制定防制措施,以免危及动物及人类健康.     

Haemaphysalis longicornis tick bites are a possible cause of red meat allergy in Japan

Recent studies revealed that Amblyomma or Ixodes tick bites may cause red meat allergy, in which galactose‐α‐1,3‐galactose (α‐Gal) is a major IgE‐binding epitope. The incidence of red meat allergy is high in Shimane Prefecture, as is tick‐transmitted Japanese spotted fever. Therefore, we speculated that tick bites may cause these meat allergies. The carbohydrate α‐Gal was detected in the salivary gland protein of Haemaphysalis longicornis (H. longicornis), the vector for Japanese spotted fever, by immunoblotting using anti‐α‐Gal antibody. H. longicornis salivary gland protein‐specific IgE was detected in the sera of 24 of 30 patients with red meat allergies. Sensitization to tick salivary gland protein containing α‐Gal is possibly a major etiology of red meat allergy; the carbohydrate plays a crucial role in its allergenicity. These results further indicate that the α‐Gal epitope is present not only in Amblyomma or Ixodes, but also in Haemaphysalis.     

长角血蜱、嗜群血蜱经期传播莱姆病螺旋体Borrelia garinii的实验研究

目的　探讨长角血蜱、嗜群血蜱经期传播莱病螺旋体的可能性。方法　通过皮下注射KM鼠建立实验感染动物模型 ,以此阳性感染鼠感染试蜱非感染种群 ,观察试蜱的感染能力以及以莱姆病螺旋体的保持能力。结果　通过剌叮阳性KM鼠 ,长角血蜱、嗜群血蜱幼蜱饱血后分别获得 6 1 3%、75 0 %的阳性感染 ,但所感染的螺旋体在感染后 2d即死亡消解 ,不能重新获得分离 ,PCR扩增阳性也只能持续到饱血后 8d ,幼蜱蜕化为若蜱后 ,所有检测结果均为阴性 ;这两种蜱的若蜱也都可通过吸血获得 70 0 %的检测阳性率 ,感染后长角血蜱、嗜群血蜱可分别在在饱血后 5、10d内保持螺旋体的活性 ,PCR检测阳性率可延迟至饱血后 15d。此后直至蜕化成为成蜱 ,所有血、蜱检测结果均呈阴性 ;长角血蜱、嗜群血蜱的不同种群在感染和保持螺旋体能力上表现一致。结论　长角血蜱、嗜群血蜱的幼蜱和若蜱虽可以感染莱姆病螺旋体但不能经期传播到下一发育阶段 ,不具备经期传播能力 ,作为莱姆病主要媒介的可能性不大。     

三种硬蜱季节消长研究

在山西省晋中太行山地带,进行了日本血蜱、长角血蜱和森林革蜱季节动态调查研究,查清了三种硬蜱成虫、若虫及幼虫活动季节及对不同宿主选择,为消灭三种硬蜱提供了理论依据。     

北京西山地区长角血蜱及其体内伯氏疏螺旋体实验观察

目的 :实验观察长角血蜱及其体内伯氏疏螺旋体形态结构。方法 :1994～ 1996年在北京西山地区采集长角血蜱 180只。标本应用蜱整体封片法、冰冻切片和石蜡切片结合HE染色和硝酸银染色法分别检测和光镜观察。结果 :蜱整体封片法直接镜检即可观察到蜱外部基本形态结构。HE染色观察蜱内部器官细微形态结构。冰冻切片和石蜡切片结合硝酸银染色 ,检出北京西山地区长角血蜱 (包括幼蜱、若蜱、成蜱和饱血蜱 )伯氏疏螺旋体感染率为 2 3 % ,显微镜下可清晰观察到蜱体伯氏疏螺旋体多种不同的形态。结论 :上述方法对莱姆病媒介蜱及伯氏疏螺旋体的流行病学研究有一定价值     

Borrelia burgdorferi Sensu Stricto DNA in Field-Collected Haemaphysalis longicornis Ticks,Pennsylvania, United States

We collected questing Haemaphysalis longicornis ticks from southeastern counties of Pennsylvania, USA. Of 263 ticks tested by PCR for pathogens, 1 adult female was positive for Borrelia burgdorferi sensu stricto, yielding a 0.4% infection rate. Continued monitoring of this invasive tick is essential to determine its public health role.     

Severe Fever with Thrombocytopenia Syndrome Virus in Ticks Collected from Humans,South Korea, 2013

We investigated the infection rate for severe fever with thrombocytopenia syndrome virus (SFTSV) among ticks collected from humans during May–October 2013 in South Korea. Haemaphysalis longicornis ticks have been considered the SFTSV vector. However, we detected the virus in H. longicornis, Amblyomma testudinarium, and Ixodes nipponensis ticks, indicating additional potential SFTSV vectors.     

吉林省延边地区自然生境内蜱种类的调查

目的调查吉林省延边地区自然生境内蜱的种类和分布。方法 2014年5—7月在延边地区龙井市、珲春市、延吉市3个市的6个调查位点用布旗法采集游离蜱类样本,显微镜下依据经典蜱类形态学种属鉴定蜱的种类。结果共采集的蜱280只,隶属2属2种,其中长角血蜱占94.64%(265/280)、全沟硬蜱占5.36%(15/280)。龙井市采集的86只蜱中,长角血蜱和全沟硬蜱的比例分别为83.72%(72/86)和16.28%(14/86);珲春市采集的187只蜱中,长角血蜱和全沟硬蜱的比例分别为99.47%(186/187)和0.53%(1/187);延吉市采集的7只蜱全部是长角血蜱。结论延边地区自然生境内主要以长角血蜱和全沟硬蜱为主,长角血蜱是优势蜱。