Abstract
ABSTRACTCrocodiles have been hunted and consumed for centuries for skins, nutrients, and medicines. These indomitable trends have overpowered restrictions from wildlife and conservation agencies, continuing the illegal trades of crocodiles across the world. This paper described the development of a very stable, fast, and secured polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay for the confirmed detection of Crocodylus porosus under any matrices and decomposing treatments. Two very short-sites (77 and 127-bp) of atp6 and cytb genes of C. porosus were controlled digested with AciI enzyme; producing distinctive RFLP patterns (83, 54, 44 & 23 bp). The enzyme digested assay was stable following extreme boiling, autoclaving, and microwaving treatments that break down DNA. The sensitivity was tested and validated in model meatballs and it was suitable for detecting 0.01% crocodile meatball matrices. The optimized RFLP assay was used to screen 3 commercial meatballs and 21 traditional medicines (TM). While no crocodile DNA was found in commercial chicken meatballs, 4/21 TM products were found correctly labelled to contain C. porosus DNA. The novel assay demonstrated sufficient merit to be used by regulatory agencies for any forensic and/or archaeological identification of C. porosus even under the state of decomposition.
Highlights
In 2016, the estimated global crimes on environment were USD 91–259 billion, threatening governments’ revenues, legitimate businesses, sustainable development, peace, and security.[1]
traditional medicines (TM) preparations often involve decoction method to consume the essences of the materials and it makes the release of DNA from background matrices very difficult.[35,36]
Crocodylus porosus is one of 23 Crocodylus species, and despite being protected, it has been overexploited for its various organs, such as meat and skins, which are believed to have healing properties for uses in exotic food products and revitalizing TM
Summary
In 2016, the estimated global crimes on environment were USD 91–259 billion, threatening governments’ revenues, legitimate businesses, sustainable development, peace, and security.[1]. Tensen[6] highlighted the four main reasons of increasing demand for wildlife: the uses of animal products in TM; superior taste and nutritive quality of wild meat as compared to captive breeds; unavailability and the growing exotic pet industry. In this regard, the increasing demand of some reptile meats, such as crocodiles, caimans, alligators, iguanas, and turtles has resulted in the development of
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