背景:按蚊群分布在整个东方和古北地区,可以传播疟疾等疾病,日本脑炎病毒,丝虫病。这项调查标志着首次进行全面研究,以对伊朗东北地区的这种疟疾媒介群的成分进行系统发育分析,与来自伊朗和世界各地不同地区的记录事件并列。
方法:从2023年4月至12月,使用各种方法从Golestan省的9个不同地点收集了蚊子。使用有效的分类键对收集的蚊子进行形态学鉴定。使用Sambio™试剂盒分离DNA。使用Oligo7和GeneRunner设计COI和ITS2引物。用Qiagen试剂盒进行PCR和纯化。随后,使用应用生物系统3730XL测序仪在MehrMam基因中心进行测序。然后使用BioEdit分析核苷酸序列并与GenBank数据进行比对。利用Kimura2参数进行碱基置换。基于AIC和BIC标准选择DNA模型。构造了贝叶斯树和最大似然树,以及单倍型网络。使用DnaSP软件计算的分子多样性统计。
结果:在这项研究中,共收集819只成年蚊子。A.hyrcanus是第二丰富的物种,主要发现于Kalaleh和Turkman县。测序和编辑的COI和ITS2序列以特定的登录号保存在GenBank中。使用ML进行系统发育分析,BI,和NJ方法证实了An的单系谱系。hyrcanus与强大的支持。伊朗的分子分析。hyrcanus发现了11种不同的单倍型,COI基因显示低多样性。ITS2基因揭示了两个分支-一个与伊朗有关,欧洲,和亚洲;另一个起源于伊朗西南部。单倍型网络显示两个主要群体-一个来自伊朗西南部,另一个来自伊朗北部。伊朗表现出六种不同的单倍型,而土耳其展示了最高的多样性。
结论:An.伊朗西南部的hyrcanus展示了一个独特的单倍群,提示可能的亚种分化。需要更多的研究来验证这种现象。
BACKGROUND: The Anopheles hyrcanus group is distributed throughout the Oriental and Palaearctic regions and can transmit diseases such as malaria, Japanese encephalitis virus, and filariasis. This investigation marks the inaugural comprehensive study to undertake a phylogenetic analysis of the constituents of this malaria vector group in the northeastern region of Iran, juxtaposed with documented occurrences from different areas within Iran and worldwide.
METHODS: Mosquitoes were collected using various methods from nine different locations in Golestan province from April to December 2023. The collected mosquitoes were identified morphologically using valid taxonomic keys. DNA was isolated using the Sambio™ Kit.
COI and ITS2 primers were designed using Oligo7 and GeneRunner. PCR and purification were performed with the Qiagen kit. Subsequently, sequencing was carried out at the Mehr Mam GENE Center using an Applied Biosystems 3730XL sequencer. The nucleotide sequences were then analyzed and aligned with GenBank data using BioEdit. Kimura 2-parameter was Utilized for base substitutions. DNA models were selected based on AIC and BIC criteria. Bayesian and Maximum Likelihood trees were constructed, along with a haplotype network. Molecular diversity statistics computed using DnaSP software.
RESULTS: In this study, a total of 819 adult mosquitoes were collected. An. hyrcanus was the second most abundant species, predominantly found in Kalaleh and Turkman counties. The sequenced and edited
COI and ITS2 sequences were deposited in GenBank under specific accession numbers. Phylogenetic analyses using ML, BI, and NJ methods confirmed a monophyletic lineage for An. hyrcanus with strong support. Molecular analysis of Iranian An. hyrcanus found 11 diverse haplotypes, with the
COI gene displaying low diversity. The ITS2 gene revealed two clades - one associating with Iran, Europe, and Asia; the other originating from southwestern Iran. The haplotype network showed two main groups - one from southwest Iran and the other from north Iran. Iran exhibited six distinct haplotypes, while Turkey showcased the highest diversity.
CONCLUSIONS: An. hyrcanus in southwestern Iran exhibits a distinct haplogroup, suggesting possible subspecies differentiation. Additional studies are required to validate this phenomenon.