Hylobates Lar Research Articles

Hidden in the white matter: Current views on interstitial white matter neurons.

The mammalian brain comprises two structurally and functionally distinct compartments: the gray matter (GM) and the white matter (WM). In humans, the WM constitutes approximately half of the brain volume, yet it remains significantly less investigated than the GM. The major cellular elements of the WM are neuronal axons and glial cells. However, the WM also contains cell bodies of the interstitial neurons, estimated to number 10 to 28 million in the adult bat brain, 67 million in Lar gibbon brain, and 450 to 670 million in the adult human brain, representing as much as 1.3%, 2.25%, and 3.5% of all neurons in the cerebral cortex, respectively. Many studies investigated the interstitial WM neurons (IWMNs) using immunohistochemistry, and some information is available regarding their electrophysiological properties. However, the functional role of IWMNs in physiologic and pathologic conditions largely remains unknown. This review aims to provide a concise update regarding the distribution and properties of interstitial WM neurons, highlight possible functions of these cells as debated in the literature, and speculate about other possible functions of the IWMNs and their interactions with glial cells. We hope that our review will inspire new research on IWMNs, which represent an intriguing cell population in the brain.

Assessment of Gut Microbiome Variations in the Mother and Twin Infant of Captive White-handed Gibbons (Hylobates lar) Reveals the Presence of Beneficial and Pathogenic Bacteria

The white-handed gibbon (Hylobates lar) is one of the three Hylobatidae species found in the Peninsular Malaysia region. Studies on primate gut microbiota have yet to be conducted in Malaysian Hylobatidae, but previous studies have covered Cercopithecidae. Knowledge of the gut microbiota of endangered captive Malaysian Hylobatidae primates is important for health assessment and conservation management. Thus, the main objective of this study was to determine the gut microbiota profile of captive white-handed gibbons using the metabarcode 16S rRNA gene. Two H. lar fecal samples collected from the mother and twin infant at Zoo Melaka were used for DNA extraction. Next-generation sequencing (NGS) of the 16S rRNA gene was performed. The NGS data were analyzed on the basis of amplicon sequence variation. The gut microbiota of H. lar was dominated by Firmicutes and Bacteroidota because of their important roles in fermentation and nutrient assimilation from plant-based food sources. Predominance of Prevotella in the gut microbiota of H. lar indicates that the captivity condition causes the gut microbiota of gibbons to resemble that of humans. Treponema, a pathogenic bacterium, was also detected in the mother and twin infant. Variation in the bacterial community profile between the mother and the infant indicated differences in dietary adaptations and physiological state. This study provides an overview of gibbon health levels through gut microbiome screening and can be used when considering Malaysian primate health welfare and captive management.

Active self-treatment of a facial wound with a biologically active plant by a male Sumatran orangutan

Although self-medication in non-human animals is often difficult to document systematically due to the difficulty of predicting its occurrence, there is widespread evidence of such behaviors as whole leaf swallowing, bitter pith chewing, and fur rubbing in African great apes, orangutans, white handed gibbons, and several other species of monkeys in Africa, Central and South America and Madagascar. To the best of our knowledge, there is only one report of active wound treatment in non-human animals, namely in chimpanzees. We observed a male Sumatran orangutan (Pongo abelii) who sustained a facial wound. Three days after the injury he selectively ripped off leaves of a liana with the common name Akar Kuning (Fibraurea tinctoria), chewed on them, and then repeatedly applied the resulting juice onto the facial wound. As a last step, he fully covered the wound with the chewed leaves. Found in tropical forests of Southeast Asia, this and related liana species are known for their analgesic, antipyretic, and diuretic effects and are used in traditional medicine to treat various diseases, such as dysentery, diabetes, and malaria. Previous analyses of plant chemical compounds show the presence of furanoditerpenoids and protoberberine alkaloids, which are known to have antibacterial, anti-inflammatory, anti-fungal, antioxidant, and other biological activities of relevance to wound healing. This possibly innovative behavior presents the first systematically documented case of active wound treatment with a plant species know to contain biologically active substances by a wild animal and provides new insights into the origins of human wound care.

Stubby versus stabby: A preliminary analysis of canine microwear in primates: Implication for inferring ingestive behaviors.

Molar and incisor microwear reflect aspects of food choice and ingestive behaviors in living primates and have both been used to infer the same for fossil samples. Canine microwear, however, has received less attention, perhaps because of the prominent role canines play in social display and because they are used as weapons-while outside of a few specialized cases, their involvement in diet related behaviors has not been obvious. Here, we posit that microwear can also provide glimpses into canine tooth use in ingestion. Canines of Sumatran long-tailed macaques (Macaca fascicularis), agile gibbons (Hylobates agilis), lar gibbons (Hylobates lar), Thomas' leaf monkeys (Presbytis thomasi), and orangutans (Pongo abelii), and two African great apes, bonobos (Pan paniscus) and common chimpanzees (Pan troglodytes schweinfurthii), were considered. The labial tips of maxillary canine replicas were scanned using a white-light confocal profiler, and both feature and texture analyses were used to characterize microwear surface patterning. The taxa exhibited significant differences in canine microwear. In some cases, these were consistent with variation in reported anterior tooth use such that, for example, the orangutans, known to use their front teeth extensively in ingestion, had the highest median number of microwear features on their canines, whereas the gibbons, reported to use their front teeth infrequently in food acquisition, had the lowest.

Variation of thoracolumbar vertebral morphology in anthropoid primates.

Morphological variation among extant primates in the lumbar vertebral column is well studied. However, knowledge concerning the thoracic spine, an important region responsible for supporting and facilitating movement in the upper trunk, remains relatively scarce. Consequently, our comprehension of the functional differentiation exhibited throughout the thoracolumbar vertebral column among various primate species remains constrained. In this study, we examined patterns of morphological variation in the thoracolumbar vertebral column of extant hominoids, cercopithecoids, and Ateles. We collected external shape data on 606 thoracic and lumbar vertebrae from Homo sapiens, Pan troglodytes, Hylobates lar, Macaca fuscata, Chlorocebus aethiops, Colobus guereza, Ateles geoffroyi, and A. belzebuth. Forty-four landmarks were obtained on the three-dimensional surface. Geometric morphometrics was used to quantify the centroid size and variation of the shapes of thoracic and lumbar vertebrae. Cercopithecoids exhibited greater variation in the size and shape of their thoracic and lumbar vertebrae compared to hominoids and Ateles. Although many vertebral features contributed to the observed variation throughout the thoracolumbar vertebral column within the taxon, the transverse and spinous processes exhibited relatively major contributions. Our results suggest that quadrupedal locomotion requires the functional differentiation between thoracic and lumbar vertebrae, and for hominoids, functional adaptation to orthograde posture necessitates a relatively more uniform shape of thoracic and lumbar vertebrae.

Integration-free induced pluripotent stem cells from three endangered Southeast Asian non-human primate species

Advanced molecular and cellular technologies provide promising tools for wildlife and biodiversity conservation. Induced pluripotent stem cell (iPSC) technology offers an easily accessible and infinite source of pluripotent stem cells, and have been derived from many threatened wildlife species. This paper describes the first successful integration-free reprogramming of adult somatic cells to iPSCs, and their differentiation, from three endangered Southeast Asian primates: the Celebes Crested Macaque (Macaca nigra), the Lar Gibbon (Hylobates lar), and the Siamang (Symphalangus syndactylus). iPSCs were also generated from the Proboscis Monkey (Nasalis larvatus). Differences in mechanisms could elicit new discoveries regarding primate evolution and development. iPSCs from endangered species provides a safety net in conservation efforts and allows for sustainable sampling for research and conservation, all while providing a platform for the development of further in vitro models of disease.

A map of white matter tracts in a lesser ape, the lar gibbon

The recent development of methods for constructing directly comparable white matter atlases in primate brains from diffusion MRI allows us to probe specializations unique to humans, great apes, and other primate taxa. Here, we constructed the first white matter atlas of a lesser ape using an ex vivo diffusion-weighted scan of a brain from a young adult (5.5 years) male lar gibbon. We find that white matter architecture of the gibbon temporal lobe suggests specializations that are reminiscent of those previously reported for great apes, specifically, the expansion of the arcuate fasciculus and the inferior longitudinal fasciculus in the temporal lobe. Our findings suggest these white matter expansions into the temporal lobe were present in the last common ancestor to hominoids approximately 16 million years ago and were further modified in the great ape and human lineages. White matter atlases provide a useful resource for identifying neuroanatomical differences and similarities between humans and other primate species and provide insight into the evolutionary variation and stasis of brain organization.

Cowpox in zoo and wild animals in the United Kingdom

Cowpox virus is considered to be a re-emerging zoonotic pathogen and a public health threat due to increasing numbers of cases in humans and animals in Europe over the past decade, including within the United Kingdom (UK). We present epidemiological data and diagnostic features of 27 recent, naturally occurring cowpox cases in zoo and wild animals across the UK, including the first reports of cowpox in two snow leopards (Panthera uncia), a Bengal tiger (Panthera tigris tigris), three Chilean pudus (Pudu puda), a Malayan tapir (Tapirus indicus) and a Eurasian otter (Lutra lutra), and the first reports of Orthopoxvirus infection in a lar gibbon (Hylobates lar), a Southern tamandua (Tamandua tetradactyla) and an aardvark (Orycteropus afer). This study provides a detailed overview of cowpox infections in a wide range of non-domestic animal species, presents a range of methods for diagnosis and demonstrates the value of retrospective analysis of pathology surveillance in revealing epidemiological links.

A cooperation experiment with white-handed gibbons (Hylobates lar)

Cooperative behaviors among individuals of numerous species play a crucial role in social interactions. There is a special interest in investigating the occurrence of cooperation among apes because this knowledge could also shed light on evolutionary processes and help us understand the origin and development of cooperation in humans and primates in general. Gibbons are phylogenetically intermediate between the great apes and monkeys, and therefore represent a unique opportunity for comparisons. The aim of the present study was to discover whether or not white-handed gibbons (Hylobates lar) show cooperative behaviors. In order to test for the respective behaviors, the gibbons were presented with a commonly used experimental cooperative rope-pulling task. The gibbons in this study did not exhibit cooperative behaviors during the problem-solving task. However, prior training procedures could not be fully completed, hence this project constitutes only the onset of exploring cooperative behaviors in gibbons. Additional behavioral observations revealed that the gibbons spent significantly more time “out of arm’s reach to everyone”, suggesting that they are less often involved in social interactions, than other, more cooperative primates.

Coordination within paraspinal muscles during bipedalism in humans, a white-handed gibbon, and a Japanese macaque

Coordination within paraspinal muscles during bipedalism in humans, a white-handed gibbon, and a Japanese macaque

Isochrony and rhythmic interaction in ape duetting.

How did rhythm originate in humans, and other species? One cross-cultural universal, frequently found in human music, is isochrony: when note onsets repeat regularly like the ticking of a clock. Another universal consists in synchrony (e.g. when individuals coordinate their notes so that they are sung at the same time). An approach to biomusicology focuses on similarities and differences across species, trying to build phylogenies of musical traits. Here we test for the presence of, and a link between, isochrony and synchrony in a non-human animal. We focus on the songs of one of the few singing primates, the lar gibbon (Hylobates lar), extracting temporal features from their solo songs and duets. We show that another ape exhibits one rhythmic feature at the core of human musicality: isochrony. We show that an enhanced call rate overall boosts isochrony, suggesting that respiratory physiological constraints play a role in determining the song's rhythmic structure. However, call rate alone cannot explain the flexible isochrony we witness. Isochrony is plastic and modulated depending on the context of emission: gibbons are more isochronous when duetting than singing solo. We present evidence for rhythmic interaction: we find statistical causality between one individual's note onsets and the co-singer's onsets, and a higher than chance degree of synchrony in the duets. Finally, we find a sex-specific trade-off between individual isochrony and synchrony. Gibbon's plasticity for isochrony and rhythmic overlap may suggest a potential shared selective pressure for interactive vocal displays in singing primates. This pressure may have convergently shaped human and gibbon musicality while acting on a common neural primate substrate. Beyond humans, singing primates are promising models to understand how music and, specifically, a sense of rhythm originated in the primate phylogeny.

The effects of visitors and social isolation from a peer on the behavior of a mixed-species pair of captive gibbons

Human visitors affect the behavior of captive animals, which is the so-called visitor effect. The number and behavior of visitors may influence stress-related behaviors in captive animals, such as self-scratching, yawning, and visitor-directed vigilance. A social group setting can be applied to alleviate such negative visitor effects and facilitate social behavior and interactions between individuals. In this study, we examined how the number and behavior of visitors are related to stress-related behaviors of a captive mixed-species gibbon pair comprising a yellow-cheek gibbon (Nomascus gabriellae) and a white-handed gibbon (Hylobates lar). The two gibbons were separated during the study period, and we examined whether the social isolation stimulated the visitor effect. The frequency of stress-related behaviors of the gibbons increased and the social playing between them decreased proportionally to visitor number. In the indoor enclosure, the gibbons increased their visitor-directed vigilance when visitors shouted or struck the glass partition. Our findings indicate that the number and behavior of visitors negatively affect captive gibbons and that a mixed-species social setting can help gibbons reduce visitor-induced stress. Future studies with larger sample sizes will improve the understanding of the visitor effect and the social setting in the captivity.

Spatial distribution of white-handed gibbon calls in relation to forest vertical components, Malaysia, from a perspective of forest management

The aim of this study was to develop a tool that allows for rapid assessment of arboreal animal distributions. In our acoustic triangulation method, we placed directional microphones atop two canopy towers to get the bearings of calls, allowing us to study the spatial distribution of calling locations of white-handed gibbon ( Hylobates lar ) within 2400 ha of the Pasoh Forest Reserve in Peninsular Malaysia in relation to three forest vertical components. The calling locations were analyzed in relation to a digital surface model, a digital terrain model, and canopy height (difference between the first two parameters) derived from LiDAR data. The locations where 49 calling points originated were mapped during the survey period (December 2018 to August 2019). All forest vertical components at the calling locations were significantly higher than the average values over the whole study site, which suggests that white-handed gibbons choose taller canopy trees at locations with higher elevation. Topographically convex areas (ridgetops or upper hillslopes) tended to be chosen as calling locations over concave areas (e.g., valleys, lower slopes). In addition, calling locations were concentrated in primary forest rather than in regenerating forest, where the canopy height was shorter and more structurally homogeneous than in the primary forest. Based on these results, we mapped the area of high probability for gibbon calling locations by compiling the four parameters of forest vertical components using principal component analysis. This map based on the vertical aspects of vegetation can be helpful in properly managing the forest for gibbon conservation, enabling a rapid assessment of land-use plans within a geographic area where the calling locations occur with high probability.

Movement dynamics of gibbons after the construction of canopy bridges over a park road

Abstract Gibbons (Hylobatidae) are species highly adapted to tree-top living. Thus, their movement can be compromised due to the negative impact roads have on canopy habitats. In this study, we built two single-rope artificial canopy bridges and a ladder bridge at two out of five locations where a group of white-handed gibbons (Hylobates lar) in Khao Yai National Park, Thailand were known to cross a main park road. We compared road crossing frequencies, home-range characteristics, and other ad libitum observations during the periods before and after bridge installation. After bridge construction was complete, the group took 10 weeks to use the single rope bridges to navigate over the road. During 442 group follow observation hours and 539 bridge observation hours, 131 crosses over the road were observed. The adult female usually crossed the road first, and the group showed a clear preference for the single-rope bridges over the ladder bridge (92 crossings versus 5). Gibbons crossed the road approximately once a day and crossed mostly at the bridge locations both before and after bridge construction. There were not significant changes in crossing rates from before (crossing between the tree branches and on the ground) to after bridge installation at both the places where bridges were installed (crossing using the bridges). Nonetheless, with more crossings being in the bridges than on the ground after bridge installation, crossings were presumably safer. These findings suggest that gibbons will cross a road on the ground, risking predation, encountering people, or being hit by a vehicle, but artificial canopy bridges provided a safer crossing option since gibbons no longer crossed on the road or jumped across wide gaps at the two locations where bridges were constructed. Maintaining canopy connectivity over roads using artificial bridges logically improves home range connectivity, potentially gene flow, and safety of canopy dwellers. However, connecting areas which were not previously connected should be considered carefully. The new connection could disrupt group dynamics, particularly for species that defend territories, such as gibbons.

Host species and habitats shape the bacterial community of gut microbiota of three non-human primates: Siamangs, white-handed gibbons, and Bornean orangutans.

The gut microbiome is essential for a host to digest food, maintain health, and adapt to environments. Bacterial communities of gut microbiota are influenced by diverse factors including host physiology and the environment. Many non-human primates (NHPs), which are physiologically close to humans, are in danger of extinction. In this study, the community structure of the gut microbiota in three NHPs: siamangs (Symphalangus syndactylus, Ss), Bornean orangutans (Pongo pygmaeus, Pp), and white-handed gibbons (Hylobates lar, Hl)—housed at the largest Zoo in Taiwan were analyzed. Pp and Ss were housed in the Asian tropical rainforest area, while Hl was housed in two separate areas, the Asian tropical rainforest area and the conservation area. Bacterial community diversity of Ss, indicated by the Shannon index, was significantly higher compared with that of Hl and Pp, while the richness (Chao 1) and observed operational taxonomic units (OTUs) were similar across the three species of NHPs. Host species was the dominant factor shaping the gut microbial community structure. Beta-diversity analysis including non-metric multidimensional scaling (NMDS) and unweighted pair group method with arithmetic mean (UPGMA) suggested gut bacterial communities of Hl housed in the conservation area were closely related to each other, while the bacterial communities of Hl in the rainforest area were dispersedly positioned. Further analysis revealed significantly higher abundances of Lactobacillus fermentum, L. murinus, and an unclassified species of Lactobacillus, and a lower abundance of Escherichia-Shigella in Hl from the conservation area relative to the rainforest area. The ratio of Lactobacillus to Escherichia-Shigella was 489.35 and 0.013 in Hl inhabiting the conservation and rainforest areas, respectively. High abundances of Lactobacillus and Bifidobacterium and a high ratio of Lactobacillus to Escherichia-Shigella were also observed in one siamang with notable longevity of 53 years. Data from the study reveal that host species acted as the fundamental driving factor in modulating the community structure of gut microbiota, but that habitats also acted as key determinants within species. The presence and high abundance of probiotics, such as Bifidobacterium and Lactobacillus, provide potential indicators for future diet and habitat optimization for NHPs, especially in zoological settings.

Branching patterns of the vascularization and innervation of the primate forelimb.

In this study, we investigate the branching patterns of the vascularization and innervation of the primate forelimb by performing detailed dissections of five unembalmed nonhuman primate specimens belonging to five different species, that is, rhesus macaque (Macaca mulatta), white-handed gibbon (Hylobates lar), Western gorilla (Gorilla gorilla), chimpanzee (Pan troglodytes), and bonobo (Pan paniscus). Results are compared with five embalmed human specimens (Homo sapiens), and anatomical data of previous studies on nonhuman primates are also included to provide a broader comparative framework. The results show that the overall configuration of the forelimb blood vessels and nerves of the different primate species is similar, although some apparent interspecific differences are found. In all nonhuman primates, in contrast to humans, the superficial vena basilica is absent. Moreover, in gorilla, chimpanzee, and bonobo, the superficial venacephalica is confined to the forearm. In humans, both an arteria interossea anterior and posterior are present, while in nonhuman primates, only an arteriainterossea anterior is present, which migrates to the posterior side at the level of the musculus pronator quadratus. For the innervation, the nervus (n.) medianus and n.ulnaris connect in the forearm of the gorilla and macaque. In the gibbon, the brachial plexus shows some differences in the branching pattern at the fasciculus level compared to the other primates. We conclude that the forelimb innervation branching pattern shows some minor differences between the nonhuman primate species, compared to higher plasticity in the vascularization. However, the exact functional implications of these differences still remain unclear. Therefore, more research on a broader range of primate species and sampling more specimens for each taxon are needed.

Introgression and mating patterns between white-handed gibbons (Hylobates lar) and pileated gibbons (Hylobates pileatus) in a natural hybrid zone.

Gibbons (Family Hylobatidae) are a suitable model for exploring hybridization in pair-living primates as several species form hybrid zones. In Khao Yai National Park, Thailand, white-handed gibbons (Hylobates lar) and pileated gibbons (Hylobates pileatus) are distributed parapatrically and hybridize in a narrow zone. Their phenotypic characteristics suggest limited inter-species gene flow, although this has never been assessed. To uncover the history and degree of gene flow between the two species, we studied the genetic structure of gibbons in the hybrid zone by analyzing fecal DNA samples, phenotypic characteristics, vocalizations and individuals' social status. We determined eight autosomal single nucleotide variant (SNV) loci, and mitochondrial DNA (mtDNA) and Y-chromosomal haplotypes of 72 gibbons. We compared these markers with reference types of wild pureblood white-handed gibbons (n = 12) in Kaeng Krachan National Park and pureblood pileated gibbons (n = 4) in Khao Soi Dao Wildlife Sanctuary. Autosomal genotypic analyses confirmed the various levels of mixed ancestry for several adult gibbons with or without atypical phenotypic traits in Khao Yai National Park. In some other adult gibbons, the mixed ancestry was not detected in either autosomal SNVs or their phenotypic traits but the mtDNA. Both male and female adult hybrids formed reproductive units mainly with a phenotypic pureblood partner and many of them produced offspring. Taken together, our results suggest that once hybridization occurs, white-handed-pileated-gibbon hybrids can reproduce with either parental species and that the backcrossing and thus introgression may occur in successive generations, with no drastic changes in phenotypic appearance.

Obstetric adaptation of the human pelvic inlet based on a sample of females and males matched for body size, and comparison with three species of primates that are sexually monomorphic in body size.

Human males are larger than females in body size; females are larger than males in pelvic size. Percentage pelvic sexual size dimorphism (sex with larger mean/sex with lower mean*100%) is used to infer selection for obstetric adaptation. As pelvic size is associated with body size, interpretation of percentage sexual size dimorphism necessitates control of body size. This study uses a larger sample of 221 females and 204 males to identify a subsample of females and males matched for femoral length, femoral head diameter, and clavicular length (25 females, 46 males). For five measures of the pelvic inlet, the human sample with control of body size is compared with three primate species that are sexually monomorphic in body size and give birth to newborns with higher relative body masses than those of humans: Aotus azarae (15 females, 19 males), Hylobates lar (29 females, 29 males), and Saguinus geoffroyi (28 females, 30 males). Results show that humans (112.0%) have the highest percentage sexual size dimorphism of pelvic inlet circumference: A. azarae (108.2%), H. lar (105.4%), and S. geoffroyi (103.3%). Humans have higher sexual size dimorphism of inlet circumference despite lower relative newborn body mass compared to the other species because human newborns are of larger body mass than predicted for an anthropoid with our adult female body mass. In humans, birthing of bigger babies than predicted for our body mass increases selection pressure on females for enlargement of the pelvic inlet.

Формирование эмпирических понятий у приматов разных таксономических групп

Much has been written about categorization in primates. However, there is a dearth in comparative studies on the formation of concepts and faculty of judgement by analogy (the part of analogical reasoning) in primates of various taxonomic groups. Our study is a comparative investigation of the formation of empirical concepts and faculty of judgement by analogy in semi-monkeys (ring-tailed lemurs), Old World monkeys (rhesus macaques), small apes (white-handed gibbons) and children aged 4 to 5. The formation of empirical concepts included the ability to differentiate between different object features. Four images were presented per a trial. The number of features underlying a concept did not affect the formation of concepts in lemurs and macaques while it made a difference for gibbons and children. This might be due to the use of different learning strategies during the formation of concepts. The children were faster than the non-human subjects to form concepts based on one feature due to the faculty of judgement by analogy. However, they lost their advantage when the formation of the concept was based on two features. Gibbons had big difficulties when inhibitory control was necessary for inhibiting the previous concept to form a new one. The study focused on the inductive function of abstract thinking responsible for the formation of concepts based on one or two features and the deductive function of abstract thinking responsible for faculty of judgement by analogy in primates of various taxonomic groups.

Taxonomic and genetic assessment of captive White-Handed Gibbons (Hylobateslar) in Peninsular Malaysia with implications towards conservation translocation and reintroduction programmes.

Conservation translocation and reintroduction for the purpose of repopulating and reinforcing extirpated or depleted populations has been recognised as an important conservation tool, particularly for gibbon conservation in the immediate future. Feasibility assessments involving multiple factors, including taxonomic and genetic assessment of rescued and captive gibbons, are imperative prior to translocation and reintroduction programmes. In this study, we attempt to determine the subspecies and origin of captive Hylobateslar, White-handed gibbons, from Peninsular Malaysia to assist in future translocation and reintroduction programmes. A total of 12 captive and rescued H.lar samples were analysed using the control region segment of mitochondrial DNA. Sequence analyses and phylogenetic trees constructed using neighbour-joining, maximum likelihood, Bayesian inference, and network methods congruently differentiate all 12 captive individuals used in this study from other H.lar subspecies suggesting that these individuals belong to the H.larlar subspecies. In addition, two populations of H.l.lar were observed: (1) a southern population consisting of all 12 individuals from Peninsular Malaysia, and (2) a possible northern population represented by three individuals (from previous studies), which might have originated from the region between the Isthmus of Kra, Surat Thani-Krabi depression, and Kangar-Pattani. Our findings suggest that the complete control region segment can be used to determine the subspecies and origin of captive H.lar.