Using DNA metabarcoding and direct behavioural observations to identify the diet of proboscis monkeys (Nasalis...

Characterizing the feeding ecology of threatened species is essential to establish appropriate conservation strategies. We focused our study on the proboscis monkey (Nasalis larvatus), an endangered primate species which is endemic to the island of Borneo. Our survey was conducted in the Lower Kinabatangan Wildlife Sanctuary (LKWS), a riverine protected area that is surrounded by oil palm plantations. We aimed to determine the diet of multiple proboscis monkey groups by using two methods. First, we conducted boat-based direct observations (scan and ad libitum sampling) and identified 67 plant species consumed by the monkeys at their sleeping sites in early mornings and late afternoons. Secondly, we used the DNA metabarcoding approach, based on next-generation sequencing (NGS, MiSeq Illumina) of faecal samples (n = 155), using the short chloroplast sequence, the trnL (UAA) P6 loop. In addition, we built a DNA reference database with the local plants available in the LKWS. When combining feeding data from both methods, we reported a diverse dietary ecology in proboscis monkeys, with at least 89 consumed plant taxa, belonging to 76 genera and 45 families. Moreover, we were able to add 22 new genera as part of the diet of this endangered colobine primate in the LKWS. The two methods provided congruent and complementary results, both having their advantages and limitations. This study contributed to enhance the knowledge on the feeding ecology of proboscis monkeys, highlighting the significance of several plant species that should further be considered in habitat restoration plans or corridor establishment.