Sequence Analysis of Mitochondrial DNAs of 12S rRNA, 16S rRNA, and Cytochrome Oxidase Subunit 1(COI) Regions in Slow Lorises (Genus Nycticebus) May Contribute to Improved Identification of Confiscated Specimens

Abstract

The slow loris (Nycticebus) is a prosimian that is popular among exotic pet lovers. In Japan, many slow lorises have been imported illegally. Prosimians that have been confiscated in raids are protected in Japanese zoos, and the number of such animals has increased. In most cases, the country of origin remains unknown and even the species can be difficult to identify from the animal's physical appearance alone. We have attempted to resolve this problem by using DNA analysis. DNA samples of five species, consisting of the Pygmy slow loris (Nycticebus pygmaeus), Bengal slow loris (Nycticebus bengalensis), Sunda slow loris (Nycticebus coucang), Javan slow loris (Nycticebus javanicus), and Bornean slow loris (Nycticebus menagensis), were extracted, amplified, and the nucleotide sequences of mitochondrial 12S rRNA, 16S rRNA, and the cytochrome oxidase subunit 1(COI) regions were compared. Differences of nucleic acid sequences of representative individuals were demonstrated.

Rediscovery of Nycticebus coucang insularis Robinson, 1917 (Primates: Lorisidae) at Tioman Island and its Mitochondrial Genetic Assessment

“…In general, two major groups were observed. The basal group was formed by N. pygmaeus that diverged earlier from the other Nycticebus species, corresponding to previous studies (Chen et al 2006(Chen et al , 2004Somura et al 2012). The second group consisted of the remaining Nycticebus species (N. bengalensis, N. coucang, N. javanicus and N. menagensis) which were previously ranked at subspecies level within N. coucang.…”

“…In BEAST v2.0, the following settings were enforced; four independent runs each with 50 million MCMC generations and sub-sampled every 50,000 generations, HKY85 substitution model with five gamma category counts (Hasegawa et al 1985), Relaxed Clock Log Normal and Birth Death Model (Stadler 2010). Gray Slender Loris (Loris lydekkerianus), Red Slender Loris (Loris tardigradus) and West African Potto (Perodicticus potto) were used as the outgroup species in phylogenetic analyses following Somura et al (2012) as these species are representatives from the major branches within the family Lorisidae.…”

Molecular data confirm the presence of Nycticebus bengalensis on Langkawi Island, Malaysia

“…The phylogenetic clade formation shown in this study is consistent with previous studies conducted on the molecular phylogeny of Nycticebus. While this study did not employ the real genetic samples of N. pygmaeus as others have done (Chen et al 2006;Somura et al 2012), we managed to produce the same tree topology in which N. pygmaeus diverged earlier as compared to N. bengalensis and N. coucang (Rovie-Ryan et al 2018). We also find support for following suggestions by Rovie-Ryan et al (2018) to utilize samples from known and credible localities only, thus increasing the confidence level of our analysis confirming the presence of N. bengalensis in Langkawi Island.…”

Molecular Phylogenetic Relationships and Unveiling Novel Genetic Diversity among Slow and Pygmy Lorises, including Resurrection of Xanthonycticebus intermedius

“…Additionally, within the N. bengalensis clade are groups of N. bengalensis samples denoting some additional genetic variation, as has been reported in previous studies [ 51 ], but unlike the Bornean groups, these do not consistently correspond to geographic areas. One group is entirely comprised of samples of the ‘dark morph’ N. bengalensis from Thailand, but the ‘northern’ N. bengalensis group includes samples from as far apart from one another as Assam, northern Burma/Myanmar, northern Vietnam, and Phu Quoc Island off the southern coast of Vietnam.…”

“…Large taxonomic and geographic gaps in coverage also impede the potential of these resources to inform wildlife trade research and management. For example, our analysis is still lacking samples from large swaths of the N. bengalensis range, as well as peninsular Thailand, southern Sumatra, and southern Borneo, which remains an issue for many rescue centers within their distribution to diagnose appropriate provenance for the release of healthy individuals back to the wild [ 51 , 80 , 81 ].…”