从自然感染犬只身上采集的狭义血红扇头蜱(蜱螨亚纲:硬蜱科)中成功分离出婴儿利什曼原虫。
Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs.
作者信息
Medeiros-Silva Viviane, Gurgel-Gonçalves Rodrigo, Nitz Nadjar, Morales Lucia Emilia D' Anduraim, Cruz Laurício Monteiro, Sobral Isabele Gonçalves, Boité Mariana Côrtes, Ferreira Gabriel Eduardo Melim, Cupolillo Elisa, Romero Gustavo Adolfo Sierra
机构信息
Programa de Pós-Graduação em Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil.
Laboratório de Parasitologia Médica e Biologia de Vetores, Universidade de Brasília, Brasília, DF, Brazil.
出版信息
BMC Vet Res. 2015 Oct 9;11:258. doi: 10.1186/s12917-015-0576-5.
BACKGROUND
The main transmission route of Leishmania infantum is through the bites of sand flies. However, alternative mechanisms are being investigated, such as through the bites of ticks, which could have epidemiological relevance. The objective of this work was to verify the presence of Leishmania spp. in Rhipicephalus sanguineus sensu lato collected from naturally infected dogs in the Federal District of Brazil.
METHODS
Ticks were dissected to remove their intestines and salivary glands for DNA extraction and the subsequent amplification of the conserved region of 120 bp of kDNA and 234 bp of the hsp70 gene of Leishmania spp. The amplified kDNA products were digested with endonucleases HaeIII and BstUI and were submitted to DNA sequencing. Isolated Leishmania parasites from these ticks were analyzed by multilocus enzyme electrophoresis, and the DNA obtained from this culture was subjected to microsatellite analyses.
RESULTS
Overall, 130 specimens of R. sanguineus were collected from 27 dogs. Leishmania spp. were successfully isolated in culture from five pools of salivary glands and the intestines of ticks collected from four dogs. The amplified kDNA products from the dog blood samples and from the tick cultures, when digested by HaeIII and BstUI, revealed the presence of L. braziliensis and L. infantum. One strain was cultivated and characterized as L. infantum by enzyme electrophoresis. The amplified kDNA products from the blood of one dog showed a sequence homology with L. braziliensis; however, the amplified kDNA from the ticks collected from this dog showed a sequence homology to L. infantum.
CONCLUSION
The results confirm that the specimens of R. sanguineus that feed on dogs naturally infected by L. infantum contain the parasite DNA in their intestines and salivary glands, and viable L. infantum can be successfully isolated from these ectoparasites.
背景
婴儿利什曼原虫的主要传播途径是通过白蛉叮咬。然而,其他传播机制也在研究中,比如蜱虫叮咬,这可能具有流行病学意义。本研究的目的是验证从巴西联邦区自然感染犬只身上采集的血红扇头蜱(广义)中是否存在利什曼原虫属。
方法
解剖蜱虫以取出其肠道和唾液腺用于DNA提取,随后扩增利什曼原虫属120bp的kDNA保守区域和234bp的hsp70基因。扩增的kDNA产物用核酸内切酶HaeIII和BstUI消化,并进行DNA测序。对从这些蜱虫中分离出的利什曼原虫寄生虫进行多位点酶电泳分析,并对从该培养物中获得的DNA进行微卫星分析。
结果
总共从27只犬只身上采集了130只血红扇头蜱样本。从四只犬只身上采集的蜱虫的五个唾液腺和肠道样本池中成功培养出了利什曼原虫属。犬血样本和蜱虫培养物扩增的kDNA产物经HaeIII和BstUI消化后,显示存在巴西利什曼原虫和婴儿利什曼原虫。通过酶电泳培养并鉴定出一株为婴儿利什曼原虫。一只犬只血液中扩增的kDNA产物与巴西利什曼原虫具有序列同源性;然而,从这只犬只身上采集的蜱虫扩增的kDNA与婴儿利什曼原虫具有序列同源性。
结论
结果证实,以自然感染婴儿利什曼原虫的犬只为食的血红扇头蜱样本在其肠道和唾液腺中含有寄生虫DNA,并且可以从这些体外寄生虫中成功分离出活的婴儿利什曼原虫。