Forensically Important Blow Flies Chrysomya pinguis, C. villeneuvi, and Lucilia porphyrina (Diptera: Calliphoridae) in a Case of Human Remains in Thailand원문보기
Monum, Tawatchai
(Department of Forensic Medicine, Faculty of Medicine, Chiang Mai University)
,
Sukontason, Kabkaew L.
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Sribanditmongkol, Pongruk
(Department of Forensic Medicine, Faculty of Medicine, Chiang Mai University)
,
Sukontason, Kom
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Samerjai, Chutharat
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Limsopatham, Kwankamol
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Suwannayod, Suttida
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Klong-klaew, Tunwadee
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
,
Wannasan, Anchalee
(Department of Parasitology, Faculty of Medicine, Chiang Mai University)
This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body o...
This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body of an unknown adult male was discovered in a high mountainous forest during winter in Chiang Mai province. The remains were infested heavily with thousands of blow fly larvae feeding simultaneously on them. Morphological identification of adults reared from the larvae, and molecular analysis based on sequencing of 1,247 bp partial mitochondrial cytochrome c oxidase subunit 1 gene (CO1) of the larvae and puparia, confirmed the above mentioned 3 species. The approving forensic fly evidence by molecular approach was described for the first time in Thailand. Moreover, neighbor-joining phylogenetic analysis of the CO1 was performed to compare the relatedness of the species, thereby affirming the accuracy of identification. As species of entomofauna varies among cases in different geographic and climatic circumstances, C. pinguis and L. porphyrina were added to the list of Thai forensic entomology caseworks, including colonizers of human remains in open, high mountainous areas during winter. Further research should focus on these 3 species, for which no developmental data are currently available.
This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body of an unknown adult male was discovered in a high mountainous forest during winter in Chiang Mai province. The remains were infested heavily with thousands of blow fly larvae feeding simultaneously on them. Morphological identification of adults reared from the larvae, and molecular analysis based on sequencing of 1,247 bp partial mitochondrial cytochrome c oxidase subunit 1 gene (CO1) of the larvae and puparia, confirmed the above mentioned 3 species. The approving forensic fly evidence by molecular approach was described for the first time in Thailand. Moreover, neighbor-joining phylogenetic analysis of the CO1 was performed to compare the relatedness of the species, thereby affirming the accuracy of identification. As species of entomofauna varies among cases in different geographic and climatic circumstances, C. pinguis and L. porphyrina were added to the list of Thai forensic entomology caseworks, including colonizers of human remains in open, high mountainous areas during winter. Further research should focus on these 3 species, for which no developmental data are currently available.
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문제 정의
Despite flies of forensic importance being known, this study was the first to confirm the presence of L. porphyrina in human remains in Thailand, especially by molecular approach, thereby listing the forensic importance of this species in the country. Its collection at the death scene in this case is comparable to that in several studies of colonized human cadavers and/or carcasses.
Conversely, cases in high mountainous habitats are rare. Therefore, this study reports a case in a high mountainous area during winter, with the aim of adding information pertaining to species of forensic importance. This is a case of blow flies (Diptera: Calliphoridae), Chrysomya pinguis (Walker), Chrysomya villeneuvi (Patton), and Lucilia porphyrina (Walker), being found on human remains, 2 of which have not been reported in Thailand before.
제안 방법
The identification of blow flies in this study was based on the morphology of larvae sampled from the corpse and adults reared from immature stages, and affirmed by molecular analysis. It should be noted that during entomological sampling from a forensic autopsy, the third instar of C.
When comparing all forensic entomology cases analyzed in Thailand since 2000, this study was the first to detect C. pinguis and L. porphyrina in human remains under tropical conditions during winter in a high mountainous area. In past forensics, C.
대상 데이터
The corpse of an unknown male was discovered on the ground in a high open mountainous area (-1,200 m asl.) in Chiang Mai province, northern Thailand (Fig. 1), during the winter of 2016. The temperature at the death scene ranged from 15 to 18˚C.
이론/모형
In addition, neighbor-joining (NJ) phylogenetic tree based on the CO1 sequences (1,213bp) of Thai blow fly species and available data from the GenBank, was constructed in the MEGA6 [6], using the Kimura 2-parameter (K2P) model with 1,000 bootstrap replicates. The CO1 sequences of Chrysomya megacephala (Fabricius) and Lucilia ampullacea (Villeneuve) which are closely related species to C.
성능/효과
porphyrina, were identified in this study. All analyzed CO1 sequences showed the highest identity of up to 99-100%; with most of them top rank hitting CO1 sequences of the Thai blow flies from Chiang Mai that were submitted previously to GenBank. Within the same species, the intraspecific divergence in CO1 of C.
The consequences of successful sequence analysis are summarized in Table 1. Based on a BLAST search, 2 genera comprised 3 species of C. pinguis, C. villeneuvi, and L. porphyrina, were identified in this study. All analyzed CO1 sequences showed the highest identity of up to 99-100%; with most of them top rank hitting CO1 sequences of the Thai blow flies from Chiang Mai that were submitted previously to GenBank.
porphyrina were observed in this study. Furthermore, the BLAST results showed non-identical (100%) CO1 sequence from GenBank, but only 99% identity of other Thai L. porphyrina isolates was hit (Table 1). To our knowledge, L.
The usefulness of CO1 barcoding as a potential identification tool was shown convincingly, splitting each fly genus into separated clade. However, CO1 was not completely distinguishable the species of L. porphyrina from L. ampullacae and sorted them into the same Lucilia clade as closely related sister species (Fig. 4). For L.
Briefly, after placing some larvae in near-boiling water, the dead larvae were then cut across the middle of the second thoracic segment (TS2) and 7th abdominal segment (AS7), using a sharp surgical blade at the 2 sites, in order to view the anterior and posterior spiracles, respectively. Incomplete peritreme, a unique characteristic of maggots in the subfamily Chrysomyinae, was found at the posterior spiracle of the hairy larvae, revealing the species as C. villeneuvi [2]. However, the creamy white and the pinkish white of non-hairy larvae contrarily possessed incomplete and complete peritremes, respectively, defining as 2 distinct unidentified species.
Investigation in this study indicated a higher abundance of non-hairy maggots, when compared to the hairy-maggot of C. villeneuvi in the current casework. However, this study observed a small number of C.
In addition, neighbor-joining (NJ) phylogenetic tree based on the CO1 sequences (1,213bp) of Thai blow fly species and available data from the GenBank, was constructed in the MEGA6 [6], using the Kimura 2-parameter (K2P) model with 1,000 bootstrap replicates. The CO1 sequences of Chrysomya megacephala (Fabricius) and Lucilia ampullacea (Villeneuve) which are closely related species to C. pinguis and L. porphyrina, respectively, were also added into the analysis, together with an outgroup sequence of a house fly, Musca domestica (Linnaeus). According to the tree (Fig.
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