Kampen H, Schöler A, Metzen M, Oehme R, Hartelt K, Kimmig P, Maier W A
Institute for Medical Parasitology, University of Bonn, Sigmund-Freud-Str, Bonn, Germany.
Exp Appl Acarol. 2004;33(1-2):93-102. doi: 10.1023/b:appa.0000029975.92510.90.
Larvae of the trombiculid mite Neotrombicula autumnalis were collected at 18 sites in and around Bonn, Germany, to be screened for infection with Borrelia burgdorferi s.l. by means of PCR. Questing larvae numbering 1380 were derived from the vegetation and 634 feeding ones were removed from 100 trapped micromammals including voles, mice, shrews and hedgehogs. In a laboratory infection experiment, a further 305 host-seeking larvae from the field were transferred onto Borrelia-positive mice and gerbils, and examined for spirochete infection at various intervals after repletion. In three cases borrelial DNA could be amplified from the mites: (1) from a larva feeding on a wild-caught greater white-toothed shrew (Crocidura russula), (2) from a pool of four larvae feeding on a B. garinii-positive laboratory mouse, and (3) from a nymph that had fed on a B. afzelii-positive laboratory gerbil as a larva. In the first case, borrelial species determination by DNA hybridization of the PCR product was only possible with a B. burgdorferi complex-specific probe but not with a species-specific one. In the second case, probing showed the same borrelial genospecies (B. garinii) as the laboratory host had been infected with. In the latter case, however, DNA hybridization demonstrated B. valaisiana while the laboratory host had been infected with B. afzelii. Subsequent DNA sequencing confirmed much higher similarity of the PCR product to B. valaisiana than to B. afzelii indicating an infection of the mite prior to feeding on the laboratory host. The negligible percentage of positive mites found in this study suggests that either the uptake of borrelial cells by feeding trombiculids is an extremely rare event or that ingested spirochetes are rapidly digested. On the other hand, the results imply a possible transstadial and transovarial transmission of borreliae once they are established in their trombiculid host. However, unless the transmission of borreliae to a given host is demonstrated, a final statement on the vector competence of trombiculid mites is not possible.
在德国波恩及其周边的18个地点采集了秋新恙螨幼虫,通过聚合酶链反应(PCR)对其进行伯氏疏螺旋体复合群(Borrelia burgdorferi s.l.)感染筛查。从植被中采集到1380只活动幼虫,从100只捕获的小型哺乳动物(包括田鼠、小鼠、鼩鼱和刺猬)身上移除了634只正在取食的幼虫。在一项实验室感染实验中,将另外305只从野外采集的寻找宿主的幼虫转移到感染了伯氏疏螺旋体的小鼠和沙鼠身上,并在饱食后的不同时间间隔检查其螺旋体感染情况。在三个案例中,可从螨类中扩增出伯氏疏螺旋体DNA:(1)从一只正在取食野生捕获的大齿白鼩(Crocidura russula)的幼虫中;(2)从四只正在取食感染了伽氏疏螺旋体(B. garinii)的实验室小鼠的幼虫混合样本中;(3)从一只若虫中,该若虫幼虫期取食了感染阿氏疏螺旋体(B. afzelii)的实验室沙鼠。在第一个案例中,仅使用伯氏疏螺旋体复合群特异性探针通过PCR产物的DNA杂交来确定伯氏疏螺旋体种类是可行的,而使用种特异性探针则不行。在第二个案例中,探针显示的伯氏疏螺旋体基因种(伽氏疏螺旋体)与实验室宿主感染的相同。然而,在第三个案例中,DNA杂交显示为瓦莱斯疏螺旋体(B. valaisiana),而实验室宿主感染的是阿氏疏螺旋体。随后的DNA测序证实,PCR产物与瓦莱斯疏螺旋体的相似性远高于与阿氏疏螺旋体的相似性,这表明该螨在取食实验室宿主之前就已被感染。本研究中发现的阳性螨类比例可忽略不计,这表明要么取食的恙螨摄取伯氏疏螺旋体细胞是极其罕见的事件,要么摄入的螺旋体被迅速消化。另一方面,结果表明一旦伯氏疏螺旋体在其恙螨宿主中定殖,可能存在跨龄期和经卵传播。然而,除非证明伯氏疏螺旋体能够传播给特定宿主,否则无法对恙螨的传播能力作出最终结论。