中华硬蜱、越原血蜱和青海血蜱经期传播莱姆病螺旋体的实验研究

以实验室培育的非感染中华硬蜱、越原血蜱和青海血蜱刺叮人工感染21天的阳性KM鼠,利用分离培养和PCR方法检测蜱对莱姆病螺旋体Borrelia garinii的保持能力以及体内螺旋体对敏感动物的感染能力。中华硬蜱、越原血蜱和青海血蜱非感染幼蜱均可以通过吸血获得莱姆病螺旋体, 然而,它们在饱血、消化、蜕皮和再次吸血过程中对莱姆病螺旋体的保持能力差异很大。只有中华硬蜱可以保持活的莱姆病螺旋体直至蜕化为下一发育阶段,蜕化后仍携带莱姆病螺旋体并对敏感KM鼠有感染能力。而越原血蜱和青海血蜱对莱姆病螺旋体的保持能力不能跨越消化、蜕皮阶段,蜕化后这两种血蜱不再携带莱姆病螺旋体,因此,这两种血蜱不具备经期传播能力,作为莱姆病媒介的可能性不大。中华硬蜱于幼蜱到若蜱以及若蜱到成蜱两个阶段均具备经期传播莱姆病螺旋体的能力,提示了中华硬蜱作为南方莱姆病的潜在媒介是可能的。     

森林革蜱雌蜱唾液腺的结构与变化(英文)

森林革蜱 (DermacentorsilvarumOlenev)雌蜱唾液腺由唾液腺管和大量的腺泡组成。从假头基到唾液腺末端 ,唾液腺管分为三部分 ,即中央腺管、主分支腺管和小叶管。球状的腺泡分布在各级腺管上。气管和中央腺管并行。腺泡呈圆形或近圆形 ,表面呈褶皱状 ,并有细小的气管分布。饥饿雌蜱唾液腺长度短 (5 4 7 3 3 μm) ;吸血后长度增加 ,吸血后 3天达到最大值 (1 1 0 9 40 μm)。从吸血后 3天到饱血后前 3 天 ,无明显变化 ,饱血后 4天明显缩短 (5 0 0 0 0 μm)。饥饿雌蜱的腺泡直径短 (45 2 4 μm) ,吸血期逐渐增大 ,吸血后 5天达到最大值 (74 1 0 μm)。饱血后腺泡逐渐萎缩并于饱血后 4天退化。     

草原革蜱和森林革蜱幼虫的形态学研究

将采自不同地区的草原革蜱Dermucentor nuttalli Olenev和森林革蜱Dermaeentor silvarumOlenev成虫在实验室中培养,对所获幼虫的形态特征用生物统计方法进行分析。结果证明,革蜱幼虫的一些形态变异很大,同一种革蜱采自不同地区或不同雌虫所产的幼虫的个体变异甚至比种间变异还大。所以,前人根据个别地区少量材料所描述的上述幼虫形态特征及所做的检索表,不能广泛适用。作者分析了大量材料后,找出上述两种幼虫较稳定的鉴别特征。根据假头宽和盾板长可以区分草原革蜱和森林革蜱;按照肩毛(Sc)和第一对背中毛(Cd₁)的长度可以把上述两种幼虫与我国北部分布的其他三种革蜱(边缘革蜱、银盾革蜱和网纹革蜱)区别开来。     

新疆四种优势种革蜱超微结构观察

【目的】明确新疆优势种革蜱超微结构鉴别特征,以期准确鉴别媒介蜱。【方法】本研究以新疆疫区4种优势种革蜱——边缘革蜱D.marginatus、草原革蜱D.nuttalli、森林革蜱D.silvarum和银盾革蜱D.niveus为实验材料,在体视显微镜下观察并鉴定物种后,进行扫描电镜观察。【结果】银盾革蜱、森林革蜱、草原革蜱和边缘革蜱体躯大小、盾板珐琅彩存在差异;银盾革蜱和边缘革蜱的肢节Ⅳ(股节、胫节和后跗节)腹面都具有3个齿状突,而草原革蜱的3个齿状突位于胫节和后跗节,森林革蜱仅后跗节有3个齿状突。此外,这4种优势种革蜱须肢腹面内侧缘刚毛排列、爪垫长度与爪长度之间的比例等特征也存在明显差异。【结论】银盾革蜱、森林革蜱、草原革蜱及边缘革蜱以肢节Ⅳ腹面齿状突和气门板为鉴别特征,结合须肢腹面内侧缘刚毛排列、爪垫长度与爪长的比例等特征,可快速准确鉴别,对新疆地区蜱传疾病综合防控具有应用意义。     

一种收集蜱类唾液的方法

建立了一种收集蜱类唾液的方法。注射多巴胺(20mgmL)10μL于森林革蜱DermacentorsilvarumOlenev吸血雌蜱血腔内,可使唾液腺分泌唾液,用毛细管连续收集30min,可得唾液15～20μL蜱。     

长角血蜱不同发育期盾窝超微结构的比较研究

为阐明蜱类盾窝及其发育特点,用扫描电镜观察了长角血蜱Haemaphysalis longicornis不同发育期盾窝的结构,并分析了血餐对盾窝发育的影响.结果表明：幼蜱仅具1对盾窝原基,且每个盾窝原基有1个盾窝孔;若蜱盾窝有了一定的发育,面积（长径×短径）增大且盾窝孔数增多（2~6个）;成蜱盾窝面积最大,且盾窝孔数达21~35个.盾窝的发育主要在幼蜱蜕皮阶段及若蜱的吸血和蜕皮阶段,雌蜱盾窝孔径显著大于雄蜱（P<0.01）,成蜱、若蜱和幼蜱的盾窝孔孔径在吸血过程中（交配雌蜱除外）各虫期均无显著变化 （P>0.05）.综合分析成蜱与未成熟蜱盾窝孔径,发现它们之间无显著差异 （P>0.05）,这在一定程度上说明蜱类的盾窝孔径在未成熟期可能已经有了雌雄分化.     

辽宁省蜱类的生态地理分布

辽宁省已知的蜱类共13种, 分属于2科6属.本文依据不同地理景观区内蜱的种类与数量, 并结合其不同生境类型, 对蜱类的生态地理分布问题作了讨论.四个地理景观区内蜱种类与数量情况如下:(1)黄土丘陵台地干草原与西辽河风沙草原区:黄土丘陵台地干草原区蜱种类少, 但数量多.草原血蜱为优势种, 草原硬蜱数量较少.西辽河沙地草原区有草原血蜱、草原革蜱、草原硬蜱, 前两种均为优势种.(2)辽西山地疏林灌从区:本区有森林革蜱、长角血蜱、全沟硬蜱及日本锐缘蜱, 前两种为优势种.(3)辽东山地森林及林缘草地区:本区蜱种较多, 林区以日本血蜱、嗜群血蜱、全沟硬蜱为多见;林缘草地以长角血蜱、森林革蜱为优势种, 另外还有钝跗硬蜱、血红扇头蜱.(4)平原丘陵农区:木区蜱种虽然不少, 但分布局限, 数量小.有长角血蜱、草原血蜱、森林革蜱、微小牛蜱, 波斯锐缘蜱.     

草原革蜱生理龄期的观察

草原革蜱Dcrmacentor nuttalli分布于我国新疆、内蒙等地.苏联、蒙古亦有发现,国内外均巳证实是蜱媒斑点热的传播媒介.采用目测、解剖、切片三种方法,进行了生理龄期观察,目测法是观测蜱的腹部厚度变化,以体侧缘为纵轴线比较腹面与此线构成的距离,分成4个生理龄期;解剖法的主要指征是蜱中肠支囊体积和形态变化,马氏管内鸟嘌呤含量和支气管束等形态的变化;切片法的主要指征是中肠上皮细胞形态变化和细胞内血红蛋白颗粒和血红素的变化.三种方法比较,结果基本一致,误差在10％左右.草原革蜱在新疆阿勒泰地区自3月下旬至5月上旬绝大部分为Ⅱ、Ⅲ期的强壮蜱,此时正值当地牧放季节,处于活动高峰的媒介蜱,具有重要的流行病学意义.     

华硬蜱和二棘血蜱的交叉免疫反应

本文首次比较了经中华硬蜱(Ixodes sinensis)叮咬三次后再经二棘血蜱(Haimaphysalis bispinosa)叮咬的家兔与仅经二棘血蜱叮咬的家兔的交叉免疫抗性。二棘血蜱叮咬被中华硬蜱致敏的家兔时,吸血增重为：143.12±32.67mg,但二棘血蜱在正常家兔体上寄生,初次吸血增重为：181.30±44.35mg,两者之间有显著性差异(P<0.01)。中华硬蜱和二棘血蜱唾液腺提取物(SGE)经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),显示两者分别有24条和22条电泳带,中华硬蜱主带有6条,分子量分别为142、105、94、66/65、64和56kD,而二棘血蜱主带有5条,分子量分别为：215、114、105、66/65和58kn,经中华硬蜱叮咬致敏的家兔血清和经二棘血蜱叫‘咬致敏的家兔血清作免疫印渍,均显示出105kD这一电泳带。该实验表明中华硬蜱和二棘血蜱叮咬家兔两者之间存在着交叉免疫反应,提示105kD蛋白质抗原可能是两者的共同抗原。     

性信息素2，6-二氯酚在长角血蜱交配行为中的作用

长角血蜱Haemaphysalis longicornis的交配行为包括7个时期,行为的完成依赖于性信息素的调节。生物测定表明：雄蜱的行为反应受雌蜱分泌的性信息素影响。堵塞雌蜱盾窝其行为受到抑制,点滴2,6-DCP或雌蜱盾窝腺提取物则被恢复。用气相色谱法测定了雌蜱盾窝腺中2,6-DCP的含量;吸血后1～２天含量最高(11.12 ng/只);吸血后3～５天即交配前下降交维持在一较恒定的水平;吸血后6～７天即交配后明显降低;饱血后检测不到2,6-DCP。2,6-DCP是长角血蜱性信息素的一种成分。     

The role of medium-sized mammals as reservoirs of Borrelia burgdorferi in southern New York

The ability of raccoons (Procyon lotor), striped skunks (Mephitis mephitis) and opossums (Didelphis virginiana) to serve as reservoirs of Borrelia burgdorferi, the spirochetal agent of Lyme disease, was compared with that of white-footed mice (Peromyscus leucopus). Twenty-eight (28) medium-sized mammals and 34 white-footed mice were captured in Westchester County, New York (USA) in summer 1986. Animals were caged over pans of water for 1 to 2 days to recover engorged tick larvae (Ixodes dammini) that detached from the hosts after feeding. With the exception of mice, numbers of engorged tick larvae recovered exceeded those counted during initial examinations of the hosts by 30% (opossums) to nearly 90% (raccoons). Newly-molted nymphal ticks derived from the engorged larvae were examined for the presence of spirochetes by darkfield microscopy. Percentage infection was 5% (n = 22) for ticks from skunks and 14% (n = 191) for ticks from raccoons. None of 24 nymphs from larvae that fed on opossums survived long enough for spirochete examination. By comparison, 40% (n = 72) of nymphs from larvae which fed on white-footed mice were infected. Of the individual hosts from which molted nymphs had fed as larvae, 67% of mice, 33% of skunks, and 55% of raccoons produced spirochete-positive ticks.     

Ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum ticks to acquire and transstadially transmit Borrelia garinii

In China, the ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum to transmit Borrelia spirochetes was determined under laboratory conditions. Results showed that all three tick species can acquire spirochetes by feeding on infected mice. However, the capability of the three species to maintain spirochetes was different. Only I. persulcatus is able to maintain spirochetes alive during molting, and subsequent tick stages transmitted the infection to naive mice. In H. concinna and D. silvarum ticks, spirochetes could not survive after the digestion period of blood and after the molting process was finished, spirochetes were no longer detected. Contrary to H. concinna and D. silvarum, I. persulcatus should be considered the principal vector of Lyme disease in north China.     

Coinfection with Borrelia burgdorferi sensu stricto and Borrelia garinii alters the course of murine Lyme borreliosis

Ixodes ricinus ticks and mice can be infected with both Borrelia burgdorferi sensu stricto and Borrelia garinii. The effect of coinfection with these two Borrelia species on the development of murine Lyme borreliosis is unknown. Therefore, we investigated whether coinfection with the nonarthritogenic B. garinii strain PBi and the arthritogenic B. burgdorferi sensu stricto strain B31 alters murine Lyme borreliosis. Mice simultaneously infected with PBi and B31 showed significantly more paw swelling and arthritis, long-standing spirochetemia, and higher numbers of B31 spirochetes than did mice infected with B31 alone. However, the number of PBi spirochetes was significantly lower in coinfected mice than in mice infected with PBi alone. In conclusion, simultaneous infection with B. garinii and B. burgdorferi sensu stricto results in more severe Lyme borreliosis. Moreover, we suggest that competition of the two Borrelia species within the reservoir host could have led to preferential maintenance, and a rising prevalence, of B. burgdorferi sensu stricto in European I. ricinus populations.     

Arthropod- and host-specific Borrelia burgdorferi bbk32 expression and the inhibition of spirochete transmission

Antisera to BBK32 (a Borrelia burgdorferi fibronectin-binding protein) and BBK50, two Ags synthesized during infection, protect mice from experimental syringe-borne Lyme borreliosis. Therefore, B. burgdorferi bbk32 and bbk50 expression within Ixodes scapularis ticks and the murine host, and the effect of BBK32 and BBK50 antisera on spirochetes throughout the vector-host life cycle were investigated. bbk32 and bbk50 mRNA and protein were first detected within engorged ticks, demonstrating regulated expression within the vector. Then bbk32 expression increased in mice at the cutaneous site of inoculation. During disseminated murine infection, bbk32 and bbk50 were expressed in several murine tissues, and mRNA levels were greatest in the heart and spleen at 30 days. BBK32 antisera protected mice from tick-borne B. burgdorferi infection and spirochete numbers were reduced by 90% within nymphs that engorged on immunized mice. Moreover, 75% of these ticks did not retain spirochetes upon molting, and subsequent B. burgdorferi transmission by adult ticks was impaired. Larval acquisition of B. burgdorferi by I. scapularis was also inhibited by BBK32 antisera. These data demonstrate that bbk32 and bbk50 are expressed during tick engorgement and that BBK32 antisera can interfere with spirochete transmission at various stages of the vector-host life cycle. These studies provide insight into mechanisms of immunity to Lyme borreliosis and other vector-borne diseases.     

Refractoriness of the western fence lizard (Sceloporus occidentalis) to the Lyme disease group spirochete Borrelia bissettii

The western fence lizard, Sceloporus occidentalis, is refractory to experimental infection with Borrelia burgdorferi sensu stricto, one of several Lyme disease spirochetes pathogenic for humans. Another member of the Lyme disease spirochete complex, Borrelia bissettii, is distributed widely throughout North America and a similar, if not identical, spirochete has been implicated as a human pathogen in southern Europe. To determine the susceptibility of S. occidentalis to B. bissettii, 6 na?ve lizards were exposed to the feeding activities of Ixodes pacificus nymphs experimentally infected with this spirochete. None of the lizards developed spirochetemias detectable by polymerase chain reaction for up to 8 wk post-tick feeding, infected nymphs apparently lost their B. bissettii infections within 1-2 wk after engorgement, and xenodiagnostic L. pacificus larvae that co-fed alongside infected nymphs did not acquire and maintain spirochetes. In contrast, 3 of 4 na?ve deer mice (Peromyscus maniculatus) exposed similarly to feeding by 1 or more B. bissettii-infected nymphs developed patent infections within 4 wk. These and previous findings suggest that the complement system of S. occidentalis typically destroys B. burgdorferi sensu lato spirochetes present in tissues of attached and feeding I. pacificus nymphs, thereby potentially reducing the probability of transmission of these bacteria to humans or other animals by the resultant adult ticks.     

Seasonal abundance and activity of the tick Dermacentor silvarum in northern China

Studies on the seasonal abundance and activity of larvae, nymphs and adults of Dermacentor silvarum (Ixodida: Ixodidae), both on and off the host, were carried out over a 2-year period from February 2008 to January 2010 in northern China. During the study period, feeding ticks were removed weekly from domestic sheep and free-living ticks were collected weekly by flag-dragging in four different habitat types. Rodents were captured and inspected for immature ticks from June to September 2008. The results of this study suggest that the ticks mainly reside in shrubs and complete only one generation per year with sequential seasonal distribution and little overlap between the activity of the larvae and nymphs. Adults were most prevalent from late February to late May and peaked in number in mid-April. Larvae were found from early June to early September and peaked in number in mid-July. Nymphs were mainly distributed from late June to late September and peaked in mid-August. Adult and nymphal D. silvarum were found primarily on the ears of sheep, but no larvae were found on sheep. Additionally, an overwintering male adult population was detected on sheep after October, but no free-living adults were found by dragging.     

Vector competence of Ixodes angustus (Acari: Ixodidae) for Borrelia burgdorferi sensu stricto

The vector competence of Ixodes angustus for Borrelia burgdorferi sensu stricto (s.s.) was investigated in the laboratory. The larval progeny of female ticks from Washington State were placed on Swiss-Webster mice that had been inoculated intravenously with 10⁸ spirochetes each of a Californian isolate of B. burgdorferi. Spirochetes were detected in 6 (12%) of 50 nymphs derived from larvae that had fed on these animals. Ten nymphs from the same cohort of experimentally infected ticks were placed on each of 4 naive deer mice (Peromyscus maniculatus). One of the mice seroconverted to B. burgdorferi and spirochetes were isolated from its ear tissues 4 weeks after exposure to ticks. Further vector competence trials were conducted with I. angustus ticks from California. Larvae were fed on deer mice that had been inoculated intradermally with B. burgdorferi along with larvae of I. spinipalpis as a comparison group. There was no significant difference in the prevalence of infection in nymphs of I. angustus (8.2%) versus those of I. spinipalpis (12.1%). We conclude that I. angustus is a competent experimental vector of B. burgdorferi s.s. and its efficiency for acquiring and transstadially passing such spirochetes is similar to that of I. spinipalpis.     

Avian and mammalian hosts for spirochete-infected ticks and insects in a Lyme disease focus in Connecticut.

Spirochetes and their vectors and reservoirs were studied in a Lyme disease focus in East Haddam, Connecticut, from mid-May through September 1983. Ixodes dammini subadults were comparable in number on white-footed mice (Peromyscus leucopus) (means = 2.9 +/- 3.6 SD) to those on 27 different species of birds (means = 2.3 +/- 4.2 SD) representing 11 families within the order Passeriformes. Less commonly found ticks on birds (means less than or equal to 0.1) were immature Ixodes dentatus and Haemaphysalis leporispalustris. Although spirochete-infected I. dammini larvae and nymphs were taken off eight and nine different species of birds, respectively, significantly fewer positive larvae were removed from birds than from white-footed mice. Spirochetes were detected in the midguts of I. dammini, Dermacentor variabilis, and H. leporispalustris and two species of insects (Cuterebra fontinella and Orchopeas leucopus). Possibly, arthropods other than I. dammini vector these spirochetes in northeastern United States. Spirochetes grew in a cell-free medium inoculated with bloods from four white-footed mice, one woodland jumping mouse (Napaeozapus insignis), one northern mockingbird (Mimus polyglottos), one gray catbird (Dumetella carolinensis), two prairie warblers (Dendroica discolor), one orchard oriole (Icterus spurius), one common yellowthroat (Geothlypis trichas), and one American robin (Turdus migratorius). We suggest that avian hosts, like mammals, develop spirochetemias of the causative agent of Lyme disease. Erythematous tissues from a white-footed mouse were infected with spirochetes.     

Association of Borrelia garinii and B. valaisiana with songbirds in Slovakia

In Europe, 6 of the 11 genospecies of Borrelia burgdorferi sensu lato are prevalent in questing Ixodes ricinus ticks. In most parts of Central Europe, B. afzelii, B. garinii, and B. valaisiana are the most frequent species, whereas B. burgdorferi sensu stricto, B. bissettii, and B. lusitaniae are rare. Previously, it has been shown that B. afzelii is associated with European rodents. Therefore, the aim of this study was to identify reservoir hosts of B. garinii and B. valaisiana in Slovakia. Songbirds were captured in a woodland near Bratislava and investigated for engorged ticks. Questing I. ricinus ticks were collected in the same region. Both tick pools were analyzed for spirochete infections by PCR, followed by DNA-DNA hybridization and, for a subsample, by nucleotide sequencing. Three of the 17 captured songbird species were infested with spirochete-infected ticks. Spirochetes in ticks that had fed on birds were genotyped as B. garinii and B. valaisiana, whereas questing ticks were infected with B. afzelii, B. garinii, and B. valaisiana. Furthermore, identical ospA alleles of B. garinii were found in ticks that had fed on the birds and in questing ticks. The data show that songbirds are reservoir hosts of B. garinii and B. valaisiana but not of B. afzelii. This and previous studies confirm that B. burgdorferi sensu lato is host associated and that this bacterial species complex contains different ecotypes.