Samerjai Chutharat, Sukontason Kabkaew L, Sontigun Narin, Sukontason Kom, Klong-Klaew Tunwadee, Chareonviriyaphap Theeraphap, Kurahashi Hiromu, Klimpel Sven, Kochmann Judith, Saeung Atiporn, Somboon Pradya, Wannasan Anchalee
Center in Insect Vector Study, Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Graduate School, Chiang Mai University, Chiang Mai 50200, Thailand.
Insects. 2019 Dec 18;11(1):2. doi: 10.3390/insects11010002.
Flesh flies (Sarcophagidae) are necrophagous insects initially colonizing on a corpse. The species-specific developmental data of the flies collected from a death scene can be used to estimate the minimum postmortem interval (PMI). Thus, the first crucial step is to correctly identify the fly species. Because of the high similarity among species of flesh flies, DNA-based identification is considered more favorable than morphology-based identification. In this study, we demonstrated the effectiveness of combined sequences (2216 to 2218 bp) of cytochrome c oxidase subunit I and II genes (COI and COII) for identification of the following 14 forensically important flesh fly species in Thailand: Boettcherisca nathani Lopes, Fengia ostindicae (Senior-White), Harpagophalla kempi (Senior-White), Liopygia ruficornis (Fabricius), Lioproctia pattoni (Senior-White), Lioproctia saprianovae (Pape & Bänziger), Parasarcophaga albiceps (Meigen), Parasarcophaga brevicornis (Ho), Parasarcophaga dux (Thomson), Parasarcophaga misera (Walker), Sarcorohdendorfia antilope (Böttcher), Sarcorohdendorfia inextricata (Walker), Sarcorohdendorfia seniorwhitei (Ho) and Seniorwhitea princeps (Wiedemann). Nucleotide variations of Thai flesh flies were evenly distributed throughout the COI-COII genes. Mean intra- and interspecific variations ranged from 0.00 to 0.96% and 5.22% to 12.31%, respectively. Using Best Match (BM) and Best Close Match (BCM) criteria, identification success for the combined genes was 100%, while the All Species Barcodes (ASB) criterion showed 76.74% success. Maximum Likelihood (ML) and Bayesian Inference (BI) phylogenetic analyses yielded similar tree topologies of monophyletic clades between species with very strong support values. The achieved sequences covering 14 forensically important flesh fly species including newly submitted sequences for B. nathani, F. ostindicae and S. seniorwhitei, can serve as a reliable reference database for further forensic entomological research in Thailand and in other areas where those species occur.
麻蝇(麻蝇科)是最初在尸体上定殖的食尸性昆虫。从死亡现场采集的苍蝇的物种特异性发育数据可用于估计最短死后间隔时间(PMI)。因此,第一步关键是正确识别苍蝇物种。由于麻蝇物种之间相似度很高,基于DNA的鉴定比基于形态学的鉴定更具优势。在本研究中,我们证明了细胞色素c氧化酶亚基I和II基因(COI和COII)的组合序列(2216至2218 bp)对于鉴定泰国以下14种具有法医学重要性的麻蝇物种的有效性:纳氏麻蝇Boettcherisca nathani Lopes、东方麻蝇Fengia ostindicae (Senior-White)、肯氏麻蝇Harpagophalla kempi (Senior-White)、红角麻蝇Liopygia ruficornis (Fabricius)、帕氏麻蝇Lioproctia pattoni (Senior-White)、萨氏麻蝇Lioproctia saprianovae (Pape & Bänziger)、白头亚麻蝇Parasarcophaga albiceps (Meigen)、短角亚麻蝇Parasarcophaga brevicornis (Ho)、肥须亚麻蝇Parasarcophaga dux (Thomson)、污麻蝇Parasarcophaga misera (Walker)、羚羊麻蝇Sarcorohdendorfia antilope (Böttcher)、难解麻蝇Sarcorohdendorfia inextricata (Walker)、史氏麻蝇Sarcorohdendorfia seniorwhitei (Ho)和威氏麻蝇Seniorwhitea princeps (Wiedemann)。泰国麻蝇的核苷酸变异均匀分布于整个COI-COII基因。种内和种间平均变异分别为0.00%至0.96%和5.22%至12.31%。使用最佳匹配(BM)和最佳近似匹配(BCM)标准,组合基因的鉴定成功率为100%,而所有物种条形码(ASB)标准的成功率为76.74%。最大似然法(ML)和贝叶斯推断(BI)系统发育分析得出物种间单系类群的树形拓扑结构相似,支持值非常高。所获得的涵盖14种具有法医学重要性的麻蝇物种的序列,包括新提交的纳氏麻蝇、东方麻蝇和史氏麻蝇的序列,可为泰国及其他有这些物种分布地区的进一步法医昆虫学研究提供可靠的参考数据库。