Abstract Varanids, known as monitor lizards are traded as pet animal commodities and demand for skin trade especially in Southeast Asian countries. Given the significant demand in the international trade, accurate species identification is imperative for Varanids particularly endangered species to prevent misidentification and fraud in marketplaces as Varanids species exhibit morphological similarities. Therefore, advanced molecular technique is vital to facilitate accurate species identification especially for closely related species. Besides, few research have used cytochrome oxidase subunit I (COI) to track the genetic and phylogenetic analysis of Varanids. In this study, the COI mitochondrial DNA region was tested as a molecular marker for species identification of Malaysia monitor lizards using the DNA barcode. Two species were successfully identified from eight samples from Pantai Penyabong, Mersing, Johor which are V. salvator and V. nebulosus based on phylogenetic analysis (Neighbor-Joining, Maximum Parsimony and Bayesian Tree). This study showed that by using their relative genetic divergence and evolutionary links, DNA barcodes can aid in the species identification through molecular technique. Short sequences can be used to identify closely related species, and the possibility of species misidentification among the study’s samples should be taken into account. DNA barcoding techniques should be widely practised especially in wildlife forensics to facilitate precise wildlife identification and improve the efficiencies in combatting illegal wildlife trade in Malaysia.
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