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Molecular Species Identification of Six Forensically Important Iranian Flesh Flies (Diptera)
Fahimeh Talebzadeh1, Mohammad Ali Oshaghi1, Kamran Akbarzadeh1
1Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Background:
Flesh flies (Diptera: Sarcophagidae) are considered as myiasis agents and important evidences in forensic investigations. However, their use has been restricted because, at all larval stages and female adults, morphological species identification is difficult or very challenging. This study investigated to test utility of mitochondrial cytochrome oxidase subunit I (mt-COI) sequences for differentiation of six forensically important Iranian flesh flies namely, Sarcophaga crassipalpis, S. flagellifera, S. hirtipes, S. aegyptica, S. africa and S. argyrostoma.
Methods:
Male specimens were morphologically identified to species level and then the genomic DNA of the flies were extracted and subjected to polymerase chain reaction (PCR) against mt-COI gene. The PCR products were sequenced and the obtained sequences were analyzed for the species specific restriction fragment length polymorphisms (RFLPs).
Results:
Rate of genetic variation between species was 6-10% which was enough to find restriction enzymes (RE) that were able to produce species-specific RFLP profiles. Combinations of three REs: BsrFI, RsaI and HinfI, provided diagnostic bands for identification of the six Sarcophaga species.
Conclusion:
The results of this study showed that molecular markers such as RFLPs enhancing the use of evidence from flesh flies in forensic investigation. However, lack proper restriction sites in the COI region inhibited introduction of a single restriction enzyme for easy species identification. It is recommended to apply larger part of DNA such as combination of COI and COII genes to provide better RFLP markers for species identification of flesh flies.
Insights
Molecular markers like restriction fragment length polymorphisms (RFLPs) can aid flesh fly identification in forensic investigations. A combination of three restriction enzymes successfully differentiated six forensically important Sarcophaga species.
Area of Science:
- Forensic entomology
- Molecular biology
- Taxonomy
Background:
- Flesh flies (Sarcophagidae) are crucial in forensic investigations but difficult to identify morphologically.
- Accurate identification of flesh flies is vital for determining post-mortem intervals.
- This study focuses on six forensically significant Iranian Sarcophaga species.
Purpose of the Study:
- To evaluate the utility of mitochondrial cytochrome oxidase subunit I (mt-COI) gene sequences for differentiating six Iranian Sarcophaga species.
- To develop a molecular method for reliable species identification of forensically important flesh flies.
- To assess the potential of restriction fragment length polymorphisms (RFLPs) for forensic applications.
Main Methods:
- Morphological identification of male flesh flies to species level.
- DNA extraction and polymerase chain reaction (PCR) amplification of the mt-COI gene.
- Sequence analysis and RFLP profiling using selected restriction enzymes.
Main Results:
- Genetic variation between species ranged from 6-10%, sufficient for RFLP differentiation.
- A combination of three restriction enzymes (BsrFI, RsaI, HinfI) generated species-specific RFLP profiles.
- This molecular approach successfully identified the six Sarcophaga species studied.
Conclusions:
- RFLPs based on mt-COI sequences enhance the forensic utility of flesh fly evidence.
- The limited number of restriction sites in the COI region necessitates using multiple enzymes for identification.
- Future research should explore larger DNA regions, like combining COI and COII genes, for improved RFLP markers.
