Yunnan Provincial Key Laboratory of Vector-Borne Diseases Control and Research, Yunnan Provincial Collaborative Innovation Center for Public Health and Disease Prevention and Control, Yunnan Institute of Parasitic Diseases Innovative Team of Key Techniques for Vector Borne Disease Control and Prevention (Developing), Yunnan Institute of Parasitic Diseases, Pu'er, 665099, People's Republic of China.
Department of Tropical Diseases, Faculty of Naval Medicine, Naval Medical University, Shanghai, 200433, China.
Parasit Vectors. 2022 Mar 18;15(1):94. doi: 10.1186/s13071-022-05167-0.
To develop an effective malaria vector intervention method in forested international border regions within the Greater Mekong Subregion (GMS), more in-depth studies should be conducted on local Anopheles species composition and bionomic features. There is a paucity of comprehensive surveys of biodiversity integrating morphological and molecular species identification conducted within the border of Laos and Cambodia.
A total of 2394 adult mosquitoes were trapped in the Cambodia-Laos border region. We first performed morphological identification of Anopheles mosquitoes and subsequently performed molecular identification using 412 recombinant DNA-internal transcribed spacer 2 (rDNA-ITS2) and 391 mitochondrial DNA-cytochrome c oxidase subunit 2 (mtDNA-COII) sequences. The molecular and morphological identification results were compared, and phylogenetic analysis of rDNA-ITS2 and mtDNA-COII was conducted for the sequence divergence among species.
Thirteen distinct species of Anopheles were molecularly identified in a 26,415 km border region in Siem Pang (Cambodia) and Pathoomphone (Laos). According to the comparisons of morphological and molecular identity, the interpretation of local species composition for dominant species in the Cambodia-Laos border (An. dirus, An. maculatus, An. philippinensis, An. kochi and An. sinensis) achieved the highest accuracy of morphological identification, from 98.37 to 100%. In contrast, the other species which were molecularly identified were less frequently identified correctly (0-58.3%) by morphological methods. The average rDNA-ITS2 and mtDNA-COII interspecific divergence was respectively 318 times and 15 times higher than their average intraspecific divergence. The barcoding gap ranged from 0.042 to 0.193 for rDNA-ITS2, and from 0.033 to 0.047 for mtDNA-COII.
The Cambodia-Laos border hosts a high diversity of Anopheles species. The morphological identification of Anopheles species provides higher accuracy for dominant species than for other species. Molecular methods combined with morphological analysis to determine species composition, population dynamics and bionomic characteristics can facilitate a better understanding of the factors driving malaria transmission and the effects of interventions, and can aid in achieving the goal of eliminating malaria.
为了在大湄公河次区域（GMS）的森林化国际边境地区开发一种有效的疟疾媒介干预方法，应更深入地研究当地按蚊物种组成和生物学特征。在老挝和柬埔寨边境地区进行的综合生物多样性调查，结合形态学和分子物种鉴定的研究很少。
在柬埔寨-老挝边境地区共捕获 2394 只成年蚊子。我们首先对按蚊进行形态学鉴定，然后使用 412 个重组 DNA-内部转录间隔区 2（rDNA-ITS2）和 391 个线粒体 DNA-细胞色素 c 氧化酶亚单位 2（mtDNA-COII）序列进行分子鉴定。比较了分子和形态鉴定结果，并对 rDNA-ITS2 和 mtDNA-COII 进行了系统发育分析，以研究物种间的序列差异。
在 26415 公里的 Siem Pang（柬埔寨）和 Pathoomphone（老挝）边境地区，通过分子鉴定共发现 13 种不同的按蚊。根据形态和分子同一性的比较，对柬埔寨-老挝边境地区优势种（按蚊、按蚊、按蚊、按蚊和按蚊）的本地物种组成的解释，形态鉴定的准确性最高，为 98.37%至 100%。相比之下，其他通过分子方法鉴定的物种，形态鉴定的准确性较低（0-58.3%）。rDNA-ITS2 的种间平均差异分别比种内差异高 318 倍和 15 倍。rDNA-ITS2 的条码间隙范围为 0.042 至 0.193，mtDNA-COII 的条码间隙范围为 0.033 至 0.047。
柬埔寨-老挝边境地区拥有丰富的按蚊物种。与其他物种相比，形态学鉴定对优势种的鉴定准确性更高。结合形态分析的分子方法确定物种组成、种群动态和生物学特征，可以更好地了解驱动疟疾传播的因素和干预措施的效果，有助于实现消除疟疾的目标。
