Monitoring compliance of CITES lion bone exports from South Africa

From 2008 to 2018, South Africa permitted the export of captive-bred African lion (Panthera leo) skeletons to Southeast Asia under CITES Appendix II. Legal exports rose from approximately 50 individuals in 2008 to a maximum of 1,771 skeletons in 2016, and has led to ongoing concerns over possible laundering of non-lion, multiple-source and wild-sourced bones. South Africa is required under its obligations to CITES to employ mechanisms for monitoring and reporting trade, and to limit the potential for illegal trade and laundering of lion and other large felid bones. Monitoring tools for legal trade are critical to compliance with CITES. Here we evaluate the CITES-compliance procedure implemented by South Africa for export of lion bones and identify six essential general points for consideration in the implementation of animal export quota compliance protocols. We provide specific insight into the South African lion bone export monitoring system through: i) outlining the protocols followed; ii) assessing the utility of cranial morphology to identify species; iii) evaluating skeleton consignment weight as a monitoring tool; and iv) presenting molecular (DNA) species assignment and pairwise-comparative sample matching of individuals. We describe irregularities and illicit behaviour detected in the 2017 and 2018 lion bone quotas. Notably, we report that the compliance procedure successfully identified and prevented the attempted laundering of a tiger (P. tigris) skeleton in 2018. We emphasise the utility of mixed-method protocols for the monitoring of compliance in CITES Appendix II export quota systems

DNA analyses of large pangolin scale seizures: Species identification validation and case studies

Pangolins are the mosttrafficked mammal in theworld, and all eightspecies are listed under CITESAppendix I.DNAbased wildlife forensic techniques are recognized as an important component of investigating a pangolin seizure. In particular, determining the species of pangolin in a seizure will 1) confirm the presence of pangolin to establish the legality of any trade, and 2) ensure appropriate laws are applied to theirfullest extentin a prosecution. Furthermore, valuable intelligence data, such as determining the geographic provenance of samples, can be produced through analysis of pangolin seizures. Despite the immense scale of the pangolin trade, standardized wildlife forensic techniquesfortesting pangolin seizures are in theirinfancy. To addressthis, here, we present a standardized genetic marker suitable for species identification of all eight pangolin species, and outline practical strategies for sampling large-volume pangolin scale seizures. We assessed the repeatability, reproducibility, robustness, sensitivity and phylogenetic resolution of this species identification test. Critically, the assay was tested in four wildlife forensic laboratories involved in testing pangolins. Additionally, we demonstrated the test’s utility to conduct geographic provenance analysis of Phataginus tricuspis samples. We analysed five large-volume pangolin scale seizures in Malaysia, which elucidated key targetspecies, poaching hotspots, and trafficking routes. Phataginustricuspis wasthe most commonly identified species(88.8%)from the seizure samples, and 84.3% of these P. tricuspisindividuals were likely sourced from western central Africa. We expect the im

Nucleotide sequence analysis to identify a one-step mutation in a STR DNA profile during paternity testing at locus D7S820

During routine paternity testing with AmpFℓSTR Identifiler Plus Kit, the kit failed to amplify the child’s allele at locus D7S820 leading to parent-child inconsistency with a single-step mutation. The aim was to identify possible causes of this mismatch. New singleplex primers were designed and the samples were amplified, cloned and sequenced using pJET1.2/blunt cloning vector forward and reverse sequencing primers. The amplicons were ascertained using CLC Bio Main Workbench. We confirmed the presence of allele dropout at the child’s locus. We describe a single nucleotide polymorphism (SNP), from cytosine (C) to thymine (T) in the kit primer binding site region of the alleged father’s profile. The child’s profile changed from homozygous to heterozygous showing that the commercial kit failed to amplify the allele and this was concluded to be most likely due to polymerase slippage