Primer Design for Non-Invasive Genetic Identification of West African Threatened Primates

Abstract

The Guinean Forests are renowned for their primate diversity, with nearly 30 distinct species. These forests have been identified as some of Africa's most critical primate conservation areas. However, intensive poaching pressure and habitat degradation cause severe threats to these species. The remaining primate populations have low population densities, have secretive habitats, and are wary of humans. This can make field studies and direct observation difficult. To develop a reliable, sensitive and simple molecular identification method using Polymerase Chain Reaction (PCR), we sequenced the mitochondrial D-loop Hypervariable 1 region (HVR1) of Diana monkey ( Cercopithecus diana) and Campbell monkey ( Cercopithecus campbelli). From these sequences and their orthologs within Eastern black-and-white colobus ( Colobus guereza) obtained from GenBank, we designed species-specific primers to amplify fragments of the mitochondrial D-loop HVR1 gene from faecal samples. These primers allow us to differentiate between Diana monkey ( C. diana), Campbell monkey ( C. campbelli), lesser spot-nosed monkey ( Cercopithecus petaurista), Red colobus ( Piliocolobus badius), Olive colobus ( Procolobus verus), King colobus ( C. polykomos), and Geoffroy's black-and-white colobus ( C. vellerosus), demonstrating their potential for the identification of West African threatened primates.