Monica Mwale

Forensic application of DNA barcoding for identification of illegally traded African pangolin scales

The escalating growth in illegal wildlife trade and anthropogenic habitat changes threaten the survival of pangolin species worldwide. All eight extant species have experienced drastic population size reductions globally with a high extinction risk in Asia. Consequently, forensic services have become critical for law enforcement, with a need for standardised and validated genetic methods for reliable identifications. The seizure of three tonnes of pangolin scales, believed to have originated from Africa, by Hong Kong Customs Authorities provided an opportunity for the application of DNA barcoding in identifying scales. Three mitochondrial DNA gene regions (COI, Cyt b, and D-loop) were amplified for a subsample of the confiscated material and compared with taxonomically verified references. All four African species were recovered as monophyletic with high interspecific uncorrected p-distance estimates (0.048-0.188) among genes. However, only three of four African species (Phataginus tricuspis, Phataginus tetradactyla, and Smutsia gigantea, originating from West and Central Africa) and one of four Asian species (Manis javanica from Southeast Asia) were identified among scales. Although the assignment of unknown scales to specific species was reliable, additional genetic tools and representative reference material are required to determine geographic origins of confiscated pangolin specimens.

Multiple independent reduction or loss of antifreeze trait in low Antarctic and sub-Antarctic notothenioid fishes

Abstract Antifreeze glycoprotein (AFGP) in Antarctic notothenioids presumably evolved once at the base of the notothenioid radiation in the Southern Ocean. Some species closely related to the endemic Antarctic notothenioids occur in non-freezing sub-Antarctic waters where antifreeze protection is unnecessary. We examined the antifreeze trait (phenotype and genotype) of these sub-Antarctic species to help infer their evolutionary history and origin. The status of the AFGP genotype (AFGP coding sequences in DNA) and/or phenotype (serum thermal hysteresis) varies widely, from being undetectable in Dissostichus eleginoides and Patagonotothen species from the Falkland Islands, minimal in Marion Island Paranotothenia magellanica and Lepidonotothen squamifrons from the South Sandwich and Bouvet islands, to considerable genotype in the Falkland Islands Champsocephalus esox and Marion Island Harpagifer georgianus. All low Antarctic notothenioid species examined show substantial AFGP trait. Mapping of the AFGP trait status onto ND2 phylogenetic trees of a large sampling of notothenioids revealed that AFGP trait reduction or loss occurred at least three independent times in different lineages.

A tale of the traded cat: development of a rapid real-time PCR diagnostic test to distinguish between lion and tiger bone

A rapid allelic discrimination real-time polymerase chain reaction (qPCR) assay has been developed for the confirmation of lion ( Panthera leo ) or tiger ( P. tigris ) DNA. The method includes a low cost allele specific PCR assay designed for each SNP variant where the genotype is determined by incorporation and subsequent fluorescence of FAM or HEX following PCR. The described method was able to accurately and rapidly distinguish between the two species, and all three mitochondrial regions (16S, 12S and control region) performed equally well. The proposed real-time PCR assay can be applied for the accurate confirmation of either lion or tiger DNA that could be used by law enforcement agencies around the world as a tool to monitor illegal trade of tiger bones.

Molecular systematics and taxonomic status of three latitudinally widespread nototheniid (Perciformes: Notothenioidei) fishes from the Southern Ocean

The taxonomic status of the three nototheniids, Lepidonotothen squamifrons, L. larseni and Gobionotothen marionensis from different localities in the Southern Ocean is re-evaluated at the DNA level. DNA sequence divergences and phylogenetic relationship were estimated using a combined mitochondrial (mtDNA, ND2 and COI) dataset and data for one nuclear gene (S7 intron 1). Phylogenies of both datasets had Lepidonotothen kempi nested within L. squamifrons lineage, with low sequence divergences (0% to 0.4%) between the two nominal species suggesting that they are populations of one species. Therefore, these results do not support the previous splitting of L. squamifrons into different species. Similarly, the L. larseni specimens also represented a single genetic unit (0.3% to 0.6%) with low geographic variation between Atlantic and Indian Ocean specimens, which does not support the splitting of this species into geographically restricted species. The mtDNA phylogeny clearly separated individuals of G. acuta from Kerguelen, Heard and MacDonald Islands from G. marionensis individuals into different clades, with sequence divergence of 2.9% between these clades supporting they are different species.

Re-description of two species of the cardinalfish genus Archamia (Teleostei: Apogonidae) from the Red Sea and Western Indian Ocean.

The cardinalfishes Archamia bilineata and A. pallida were originally described from a small number of specimens collected in the Gulf of Aqaba, Red Sea and Oman, respectively. Both species are re-described using specimens collected recently in Yemen and Kenya, including the first known adult of A. pallida. These new collections increased the geographical range of both species considerably. Differences between the two populations now known for each of the species are discussed.