Abstract
To support efforts in prosecuting wildlife crimes, we developed and validated a multiplex High-Resolution Melt (M-HRM) assay for the identification of proboscidean taxa commonly required to be identified or excluded in ivory seizures and forensic casework: Asian elephant (Elephas maximus), African elephant (Loxodonta spp.), mammoth (Mammuthus spp.), and mastodon (Mammut spp.). Five hundred and fifty (550) blood, tissue, and ivory samples from individuals of these 4 proboscidean taxa were used to develop and validate the 2 proboscidean-specific mitochondrial sites targeted by this assay. The 28-basepair (bp) 16S ribosomal RNA (rRNA) and 54-bp cytochrome b (Cytb) gene segments yield a combination of melt peaks that create composite melt profiles unique to each of the 4 proboscidean taxa. Wildlife forensic laboratories can use this sensitive, rapid, and cost-effective assay to assist efforts to combat the unlawful commercialization of proboscidean ivory and to stop the poaching crisis leading to the decline of these ivory-bearing species in the wild.
Highlights
Illegal poaching and wildlife trafficking is the fourth largest trans continental crime, worth an estimated $20 billion each year [1]
We have developed a multiplex High-Resolution Melt (M-HRM) assay that can distinguish between 4 proboscidean taxa protected under California’s ivory law and are required to either be identified or excluded from casework consideration – African elephant (Loxodonta spp.), Asian elephant (Elephas maximus), mammoth (Mammuthus spp.), and mastodon (Mammut spp.)
Sperm whale produced a double melt peak at cytochrome b (Cytb) that could be distinguished from target peaks, due to peak morphology and melt temperatures of 76.6 ◦C and 80 ◦C
Summary
Illegal poaching and wildlife trafficking is the fourth largest trans continental crime, worth an estimated $20 billion each year [1]. In 2015, California enacted a law (AB 96, codified as Fish and Game Code section 2022) that prohibits the purchase, sale, offer for sale, possession with intent to sell, or importa tion with intent to sell of ivory from elephant, mammoth, and mastodon as well as other non-proboscidean species [5]. It is worth noting that there are other species included in the Order Proboscidea, most notably the extinct straight-tusked elephant (Paleo loxodon antiquus), which aside from mammoth, has the most abundant fossil records in Eurasia. Even though nuclear DNA studies show the straight-tusked elephant lineage derives from lineages related to African forest elephant and mammoth [7], they have not been reported as commercialized or otherwise protected under law, and have been omitted from this study
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