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Rickettsial infection in Amblyomma cajennense ticks and capybaras (Hydrochoerus hydrochaeris) in a Brazilian spotted fever-endemic area

机译:巴西斑疹热病流行地区的cambtenams壁虱和水豚的立克次体感染

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摘要

Abstract Background Brazilian spotted fever (BSF), caused by the bacterium Rickettsia rickettsii, is the deadliest spotted fever of the world. In most of the BSF-endemic areas, capybaras (Hydrochoerus hydrochaeris) are the principal host for the tick Amblyomma cajennense, which is the main vector of BSF. Methods In 2012, a BSF case was confirmed in a child that was bitten by ticks in a residential park area inhabited by A. cajennense-infested capybaras in Itú municipality, southeastern Brazil. Host questing A. cajennense adult ticks were collected in the residential park and brought alive to the laboratory, where they were macerated and intraperitoneally inoculated into guinea pigs. A tick-inoculated guinea pig that presented high fever was euthanized and its internal organs were macerated and inoculated into additional guinea pigs (guinea pig passage). Tissue samples from guinea pig passages were also used to inoculate Vero cells through the shell vial technique. Infected cells were used for molecular characterization of the rickettsial isolate through PCR and DNA sequencing of fragments of three rickettsial genes (gltA, ompA, and ompB). Blood serum samples were collected from 172 capybaras that inhabited the residential park. Sera were tested through the immunofluorescence assay using R. rickettsii antigen. Results A tick-inoculated guinea pig presented high fever accompanied by scrotal reactions (edema and marked redness). These signs were reproduced by consecutive guinea pig passages. Rickettsia was successfully isolated in Vero cells that were inoculated with brain homogenate derived from a 3rd passage-febrile guinea pig. Molecular characterization of this rickettsial isolate (designated as strain ITU) yielded DNA sequences that were all 100% identical to corresponding sequences of R. rickettsii in Genbank. A total of 83 (48.3%) out of 172 capybaras were seroreactive to R. rickettsii, with endpoint titers ranging from 64 to 8192. Conclusions A viable isolate of R. rickettsii was obtained from the tick A. cajennense, comprising the first viable R. rickettsi isolate from this tick species during the last 60 years. Nearly half of the capybara population of the residential park was seroreactive to R. rickettsii, corroborating the findings that the local A. cajennense population was infected by R. rickettsii.
机译:摘要背景巴西立克次氏体立克次氏体细菌引起的巴西斑疹热是世界上最致命的斑疹热。在大多数BSF流行地区,水豚(Hydrochoerus hydrochaeris)是the虫Amblyomma cajennense的主要寄主,后者是BSF的主要媒介。方法2012年,在巴西东南部伊图市的一个居民公园内,有一个儿童被s虫咬伤,该儿童被tick虫咬伤,该儿童居住在该公园内。在寄宿公园收集寄主询问ca.cajennense的成年s并将其活体带到实验室,在那里将它们浸软并腹膜内接种到豚鼠中。对出现高烧的tick虱豚鼠实施安乐死,将其内部器官浸软并接种到其他豚鼠中(豚鼠传代)。来自豚鼠传代的组织样品也通过壳管技术用于接种Vero细胞。通过PCR和三个立克次体基因(gltA,ompA和ompB)片段的DNA测序,将感染的细胞用于立克次体分离物的分子表征。从居住在住宅公园中的172个水豚采集血清样本。通过使用立克次氏菌抗原的免疫荧光测定法检测血清。结果接种tick的豚鼠出现高烧并伴有阴囊反应(水肿和明显发红)。这些迹象通过连续的豚鼠繁殖而得以再现。立克次体成功地在Vero细胞中分离,该细胞接种了来自第三代高热豚鼠的脑匀浆。此立克次体分离株(指定为ITU菌株)的分子鉴定产生的DNA序列与Genbank中立克次氏体的相应序列均100%相同。在172个水豚中,共有83个(48.3%)对立克次氏菌具有血清反应活性,终点滴度范围为64至8192。结论从ca虫中获得了立克次氏菌的可行分离株,其中包括第一个可行的R。在过去的60年中,rickettsi从这种壁虱物种中分离出来。居民公园的水豚种群中近一半对立克次氏菌具有血清反应活性,从而证实了当地的A. cajennense种群被立克次氏体感染的发现。

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