Use Of Lucilia Cuprina Wiedemann (1830) Larvae For Detection Of Heroin Metabolites And Identification Of Host DNA
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Date
2016-08
Authors
Ishak, Nooratiny
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Abstract
This study investigates the capability of 3rd instar larvae of Lucilia cuprina Wiedemann (1830) in toxicology analysis to detect the heroin metabolites by using GCMS application and in molecular analysis by utilizing the Cytochrome b gene (mtDNA) for precise identification of minced buffalo meat (which had been used as experimental materials in this study) in the absence of heroin. The study conducted also investigates the effect of heroin on the Lucilia cuprina duration of life cycle. Then the developed method was applied in exact forensic cases to examine the effectiveness of the developed method in handling both human and animal forensic cases. The toxicology analysis conducted on the 3rd instar larval crop was successfully detecting a metabolite of heroin, such as morphine. The heroin that was mixed with the minced buffalo (Bubalus bubalis, Linnaeus, 1758) meat in different concentrations, accelerated the duration of life cycle of L. cuprina larvae compared to the larvae that were collected from the minced buffalo meat without heroin, which had a longer life cycle. The molecular analysis conducted on 3rd instar larval crop, succesfully amplify desired results of minced buffalo meat (with heroin) DNA sequence, which is Bubalus bubalis. The addition of heroin was found to give an impact, which reflected the nucleotide changes to the buffalo DNA sequences obtained. Eleven (11) DNA polymorphisms were detected to occur in the buffalo DNA sequences, which derived from the 3rd instar larval crop. The nucleotide changes in the buffalo DNA sequence also led to some changes in protein sequences.
The molecular analysis carried out on the 3rd instar larval crop found in actual forensic cases successfully amplify DNA sequences of Gallus gallus, Linnaeus, (1758); Sus scropa, Linnaeus, (1758); Civet (Paradoxurus hermaphrodite, Pallas, 1777); and Bubalus bubalis, Linnaeus, (1758). Besides being able to detect the animal DNA sequences in animal forensic cases, the 3rd instar larval crop found in human forensic cases also managed to determine human DNA sequence and this evidence has, in fact, been used for court proceedings. The 3rd instar larval crop was used in both cases because the 3rd instar larvae have a maximum length and all the food are stored in the 3rd instar larval crop before larvae start to pupate. The length of 3rd instar larvae, indeed, is an important criterion because at this stage, the crop can easily be seen under a microscope and dissected to remove the crop. The study, which had been conducted in forensic case, displayed that entomological evidence, especially the larval stages, could be used in identifying the host DNA in conditions where biological evidence was absent.
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The capability of 3rd instar larvae of Lucilia cuprina Wiedemann , to detect the heroin metabolites.