Illegal product manufacturing and exportation from Pakistan: Revealing the factuality of highly processed wildlife skin samples via DNA mini-barcoding

Abstract

Illegal trade is a major threat to the biodiversity and the efforts initiated for the conservation of wildlife. The shortcomings of the traditional taxonomic identification methods have been coped by a revolutionary and emerging technique, the "DNA (Deoxyribonucleic Acid) barcoding". Here we report a case of trader who was allegedly making footwear for a famous international celebrity from wild animal cutis. The samples confiscated during a raid on a footwear manufacturing industry by KP Wildlife department in August, 2016, were received by Bioresource Research Centre (BRC) for molecular identification on 1 September, 2016. The study costed about USD 88 from processing to the identification of the samples. The samples identified via DNA mini-barcoding by targeting cytochrome oxidase I (COI) gene belong to Gazella bennettii and Bos taurus. Such studies are helpful for credible investigations that only lead to effective prosecution and control of illegal wildlife trade ultimately helping in conservation of wild animals.

Single Fragment or Bulk Soil DNA Metabarcoding: Which is Better for Characterizing Biological Taxa Found in Surface Soils for Sample Separation?

“…In contrast to this, higher yields for both COI amplicons were obtained from the bulk soil extracts (Figure 2). The COI primers used in this study are universal and have been widely used in studies with diverse invertebrate and vertebrate animal taxa [35,36,37,38,39,40]. In our initial study [20], the total quantity of DNA isolated from individual insect fragments was low (0.45 ± 1.27 μg) and successful amplification of either COI region (i.e., entire or mini) was only obtained in 74% of fragments.…”

“…For clarity, amplifications of plant barcoding loci matK and rbcL were completed using the KAPA3G Plant DNA polymerase (KAPA Biosystems, Wilmington, MA, USA) and the COI barcode region was amplified using the Q5 ® Hot Start High-Fidelity DNA polymerase (New England BioLabs Inc, Ipswich, MA, USA). It is important to emphasize that while the COI primers used in Meiklejohn et al [20] amplified the appropriate region from individual insect fragments, these primer pairs are universal and contain some degenerate bases at the third codon position to permit successful amplification across a diverse range of vertebrate and invertebrate animal taxa [35,36,37,38,39,40]. A negative and appropriate positive control (either an extract of a Gerbera sp.…”

Forensic DNA Analysis

“…DNA meta-barcoding has been successfully applied to the illegal orchid trade as well as in the identification of endangered species in complex samples. − DNA mini-barcoding was utilized for the identification of highly processed animal skin and fur, wildlife skin samples, and samples of animal claws. − The forensic analysis of cytochrome b (cyt b), a mtDNA gene, has become an essential tool for species identification in routine practice, and for this reason, a number of different primers were designed with species identification capabilities. − Another study combined a morphometric approached with DNA analysis using cyt b and 16S rRNA genes to identify seized samples of tiger claws in India . Illegal trading of ivory is responsible for the decrease in elephant populations.…”

Utility of mitochondrialCOI gene for identification of wild ungulate species of conservational importance from Pakistan

“…The advancement in species identification research using molecular techniques, particularly DNA barcoding has been a great milestone in determining ungulate diversity worldwide. It has been applied to studies regarding biodiversity, phylogeny, food adulteration cases, illegal trades of ungulates and their manufactured products (Kumar et al 2017;Zhong et al 2017;Khan et al 2018;Shukla et al 2019;Bhaskar et al 2020). However, most of the ungulate diversity of Pakistan has not yet been studied through mitogenome analysis and has been restricted to the pioneer morphological studies conducted by Schaller (1980), Mirza (1998), andRoberts (1997).…”