Laboratory of Parasitology and Vector-Borne-Diseases, Institut Pasteur du Maroc, Casablanca, Morocco.
Molecular Genetics and Immunophysiopathology Research Team, Health and Environment Laboratory, Aïn Chock Faculty of Sciences, Hassan II University of Casablanca, Casablanca, Morocco.
Transbound Emerg Dis. 2022 May;69(3):1073-1083. doi: 10.1111/tbed.14064. Epub 2021 Mar 23.
In Morocco, leishmaniases are a major public health problem due to their genetic diversity and geographical distribution. Cutaneous leishmaniasis (CL) is an infectious disease caused by various species of Leishmania and transmitted typically by bite of phlebotomine sand flies. This study identifies sand fly fauna in Ibaraghen village, province of Azilal, which is a focus of CL, by combination of morphological and molecular methods (sequencing of COI gene, MALDI-TOF MS protein profiling). Nested-kDNA PCR was used to detect and identify Leishmania species within potential vector species. 432 CDC light traps were placed at different heights above ground level at four capture sites during a whole year. Traps at 1.5 m above the ground yielded capture of sand flies almost double compared to above ground level (29.33%), while the collection reached 55.09% when the traps were placed 2.5 m above ground. A total of 2,830 sand flies were collected, 2,213 unfed specimens were morphologically identified, 990 males (44.73%) and 1,223 females (55.26%) of 13 species; ten Phlebotomus species and three Sergentomyia species. Six species were analysed by MALDI-TOF MS protein profiling (4 Phlebotomus and 2 Sergentomiya species), and their identification was confirmed by COI sequencing. 1,375 unfed females were screened for the presence of Leishmania by nested-kDNA PCR in pools, 11/30 pools of P. sergenti showing a single band of 750 bp corresponding to L. tropica. Our results confirm the role of P. sergenti as a proven vector in Azilal focus of cutaneous leishmaniasis; however, the relative abundance of other species known as vectors of Leishmania species emphasizes the risk of introduction of L. infantum and L. major in this province. For the first time in Morocco, a combined approach to identify sand flies by both morphology and molecular methods based on DNA barcoding and MALDI-TOF MS protein profiling was applied.
在摩洛哥，由于其遗传多样性和地理分布，利什曼病是一个主要的公共卫生问题。皮肤利什曼病（CL）是一种由各种利什曼原虫引起的传染病，通常通过白蛉沙蝇的叮咬传播。本研究通过形态学和分子方法（COI 基因测序、MALDI-TOF MS 蛋白分析）结合鉴定了阿齐拉勒省伊巴拉根村的沙蝇区系，该地区是 CL 的一个重点。巢式 kDNA PCR 用于检测和鉴定潜在媒介物种中的利什曼原虫种。在一年中的四个采集点，在不同的离地高度设置了 432 个 CDC 轻型诱捕器。离地 1.5 米高的诱捕器捕获的沙蝇数量几乎是地面水平的两倍（29.33%），而当诱捕器放置在离地面 2.5 米高时，收集率达到 55.09%。共收集到 2830 只沙蝇，2213 只未喂食标本经形态学鉴定为 13 种中的 990 只雄性（44.73%）和 1223 只雌性（55.26%）；10 种白蛉和 3 种塞伦盖蒂亚种。通过 MALDI-TOF MS 蛋白分析（4 种白蛉和 2 种塞伦盖蒂亚种）对六种物种进行了分析，并通过 COI 测序确认了其鉴定。对 1375 只未喂食的雌性进行了巢式 kDNA PCR 检测，在 P. sergenti 的 30 个虫池中，有 11 个显示出 750 bp 的单一条带，对应于 L. tropica。我们的结果证实了 P. sergenti 作为阿齐拉勒皮肤利什曼病疫区的已证实媒介的作用；然而，其他已知作为利什曼物种媒介的物种的相对丰度强调了在该省引入 L. infantum 和 L. major 的风险。这是摩洛哥首次应用基于 DNA 条形码和 MALDI-TOF MS 蛋白分析的形态学和分子方法相结合来鉴定沙蝇。
