Functional genomics analysis reveals the evolutionary adaptation and demographic history of pygmy lorises.

Lorises are a group of globally threatened strepsirrhine primates that exhibit many unusual physiological and behavioral features, including a low metabolic rate, slow movement, and hibernation. Here, we assembled a chromosome-level genome sequence of the pygmy loris (Xanthonycticebus pygmaeus) and resequenced whole genomes from 50 pygmy lorises and 6 Bengal slow lorises (Nycticebus bengalensis). We found that many gene families involved in detoxification have been specifically expanded in the pygmy loris, including the GSTA gene family, with many newly derived copies functioning specifically in the liver. We detected many genes displaying evolutionary convergence between pygmy loris and koala, including PITRM1. Significant decreases in PITRM1 enzymatic activity in these two species may have contributed to their characteristic low rate of metabolism. We also detected many evolutionarily convergent genes and positively selected genes in the pygmy loris that are involved in muscle development. Functional assays demonstrated the decreased ability of one positively selected gene, MYOF, to up-regulate the fast-type muscle fiber, consistent with the lower proportion of fast-twitch muscle fibers in the pygmy loris. The protein product of another positively selected gene in the pygmy loris, PER2, exhibited weaker binding to the key circadian core protein CRY, a finding that may be related to this species' unusual circadian rhythm. Finally, population genomics analysis revealed that these two extant loris species, which coexist in the same habitat, have exhibited an inverse relationship in terms of their demography over the past 1 million years, implying strong interspecies competition after speciation.

CLINICAL FINDINGS OF DENTAL DISEASE AND POTENTIAL CONTRIBUTING FACTORS IN PYGMY SLOW LORISES (NYCTICEBUS PYGMAEUS) UNDER HUMAN CARE.

Dental disease is a common finding in pygmy slow lorises (Nycticebus pygmaeus) under human care, but the etiology is not fully understood. The small oral cavity in this species can make diagnosis of dental disease difficult. This retrospective study evaluated medical records and diet and husbandry protocols from 18 participating institutions with the objective of describing the signalment, clinical signs, physical exam findings, tooth type, tooth location, diagnostics used, and treatments performed to help guide care for dental disease. In addition, the study aimed to identify potential contributing factors to dental disease in this species. Of 59 animals with medical records evaluated, 42 (71.2%) had dental disease: 19 (44.2%) males, 20 (46.5%) females, and 3 (9.3%) without gender documented. Average age at onset of dental disease was 7.6 yr in males and 9 yr in females. Multiple lorises with dental disease (n = 12; 28.6%) had no premonitory clinical signs, and dental disease was found incidentally on examination. On dental examination, 30 lorises (71.4%) had evidence of gingivitis. In 13 cases skull radiographs were taken, but the majority of images (n = 8; 61.5%) were nondiagnostic for pathologic dental changes. A small proportion of cases with dental abnormalities (n = 4; 9.5%) were diagnosed using computed tomography. In total, 175 teeth were extracted from 31 patients; molars were the most frequently extracted tooth (n = 55; 31.4%). No substantial differences in diets were noted among many of the participating institutions, and not all slow lorises evaluated developed dental disease (n = 17; 28.8%). This retrospective study provides clinical findings on slow loris dental disease and guidance for the veterinary care and management of slow lorises under human care.

Welfare assessment of slow loris (Nycticebus spp.) at an Indonesian primate rehabilitation center: Development and validation of body condition score.

Body condition scoring (BCS) assessment can reflect animal welfare status and help the veterinarian to make a quick health management decision, including for confiscated slow loris (Nycticebus spp.). The confiscated slow loris should be rehabilitated in a rehabilitation center before being released. It is essential to monitor the welfare of slow loris to ensure that candidates are released. Assessment of animal welfare status requires representative measurable criteria and indicators. However, there is no standardized BCS for slow loris yet. This study focuses on developing and validating BCS based on body weight and circumference. In this study, 180 individuals were evaluated and scored. We measured body weight and circumferences to validate the assessment of BCS. There are no significant differences in body weight and circumferences within species and sexes. Muscle mass and fat deposits were palpated, visually viewed, and grouped in five BCS. There was a significant difference in body weight and circumference between BCS levels. According to this study, the development of BCS is valid and can be used to slow loris in prevailing circumstances and any ex-situ facilities.

Reducing stress and stereotypic behaviors in captive female pygmy slow lorises (Nycticebus pygmeaus).

Improving captive conditions of pygmy slow lorises (Nekaris and Nijman have recently suggested that the pygmy slow loris should be called the pygmy loris and is distinctive enough to warrant a new genus, Xanthonycticebu) (Nycticebus pygmeaus) poses many challenges because detailed aspects of their lives in the wild are incomplete. This hinders efforts to replicate sustainable environments for them. To improve their well-being in captivity, eight rescued female pygmy slow lorises at the Japan Monkey Center (JMC) were socially housed in two types of groups following their solitary housing: two pairs and one group of four individuals. They spent much of their time in affiliative behaviors, as well as sharing sleeping sites after placement in a social group. The purpose of my study was to examine whether social housing helped in reducing stress by comparing fecal glucocorticoids and stereotypic behaviors when housed alone and when with conspecifics. Overall, the levels of fecal glucocorticoids were significantly lower when socially housed than when kept alone. One individual exhibited stereotypic behavior when housed alone, but this behavior disappeared after social housing. These findings support recent evidence that pygmy slow lorises are social animals and will benefit from group housing in captivity. We conclude that social housing of pygmy slow lorises improves their well-being by reducing stress levels, and that their group housing in captivity can provide dividends for the conservation of this endangered nocturnal primate because lorises intended for release should find it easier to adapt to natural conditions.

Breaking the constraint on the number of cervical vertebrae in mammals: On homeotic transformations in lorises and pottos.

Mammals almost always have seven cervical vertebrae. The strong evolutionary constraint on changes in this number has been broken in sloths and manatees. We have proposed that the extremely low activity and metabolic rates of these species relax the stabilizing selection against changes in the cervical count. Our hypothesis is that strong stabilizing selection in other mammals is largely indirect and due to associated pleiotropic effects, including juvenile cancers. Additional direct selection can occur due to biomechanical problems (thoracic outlet syndrome). Low metabolic and activity rates are thought to diminish these direct and indirect effects. To test this hypothesis within the primates, we have compared the number of cervical vertebrae of three lorisid species with particularly low activity and metabolic rates with those of more active primate species, including with their phylogenetically closest active relatives, the galagids (bushbabies). In support of our hypothesis, we found that 37.6% of the lorisid specimens had an abnormal cervical count, which is a higher percentage than in the other nine primate families, in which the incidence varied from zero to 2.2%. We conclude that our data support the importance of internal selection in constraining evolvability and of a relaxed stabilizing selection for increasing evolvability. Additionally, we discuss that there is no support for a role of the muscularized diaphragm in the evolutionary constraint.

Social relationships among captive male pygmy slow lorises (Nycticebus pygmaeus): Is forming male same-sex pairs a feasible management strategy?

Little is known about the social behavior of pygmy slow lorises, in particular, the social relationships of same-sex individuals have rarely been investigated. The Slow Loris Conservation Center was built at the Japan Monkey Center to enhance the welfare of confiscated slow lorises, promote their conservation, improve public education, and perform scientific research on the species. In the course of improving housing conditions, several same-sex pairs of pygmy slow lorises were formed. We monitored their behaviors and fecal glucocorticoid metabolite (FGM) levels to understand whether male same-sex pairings could be a feasible management strategy. The subjects were 10 male and 6 female lorises for comparison, all of whom were over 5 years old. We successfully formed five pairs of male lorises after eight formation attempts. Male pairs initially showed some aggressive behaviors; however, the rate decreased approximately 10 days after introduction. All of the male pairs eventually exhibited extensive affiliative social behaviors, including allogrooming and social play, during the dark (active) phase, and sleep site sharing during the light (inactive) phase. The rate of sleep site sharing during the light phase was higher than expected, suggesting that the pairs preferred to stay near each other. There was no evidence of increased stress after a long period of male-male social housing. Female same-sex pairs and male-female pairs demonstrated a high level of affiliative behaviors right after the introduction. These results highlight the flexibility and high sociability of this species and indicate that such same-sex pairings are a feasible option for their social management.

Variation in predicted COVID-19 risk among lemurs and lorises.

The novel coronavirus SARS-CoV-2, which in humans leads to the disease COVID-19, has caused global disruption and more than 2 million fatalities since it first emerged in late 2019. As we write, infection rates are at their highest point globally and are rising extremely rapidly in some areas due to more infectious variants. The primary target of SARS-CoV-2 is the cellular receptor angiotensin-converting enzyme-2 (ACE2). Recent sequence analyses of the ACE2 gene predict that many nonhuman primates are also likely to be highly susceptible to infection. However, the anticipated risk is not equal across the Order. Furthermore, some taxonomic groups show high ACE2 amino acid conservation, while others exhibit high variability at this locus. As an example of the latter, analyses of strepsirrhine primate ACE2 sequences to date indicate large variation among lemurs and lorises compared to other primate clades despite low sampling effort. Here, we report ACE2 gene and protein sequences for 71 individual strepsirrhines, spanning 51 species and 19 genera. Our study reinforces previous results while finding additional variability in other strepsirrhine species, and suggests several clades of lemurs have high potential susceptibility to SARS-CoV-2 infection. Troublingly, some species, including the rare and endangered aye-aye (Daubentonia madagascariensis), as well as those in the genera Avahi and Propithecus, may be at high risk. Given that lemurs are endemic to Madagascar and among the primates at highest risk of extinction globally, further understanding of the potential threat of COVID-19 to their health should be a conservation priority. All feasible actions should be taken to limit their exposure to SARS-CoV-2.

Bioacoustics as a tool to monitor the estrus cycle in a female slow loris (Nycticebus sp.).

Evidence-based husbandry leading to increased reproductive success and strengthening of ex situ populations of slow lorises (Nycticebus spp.) living in zoos is highly important. Better fulfillment of their social needs is one of the main priorities in achieving these objectives. We performed 21-month long acoustic monitoring of a zoo-kept female slow loris (Nycticebus sp.) housed either singly or with one of two males to examine whether her estrus cycle potentially could be detected based on her vocal activity. We found a regular cycle of remarkably increased whistle production that lasted approximately 31.2 days in a nocturnal exhibit and approximately 39 days in an off-show room. The regular cycle of increased vocal activity corresponded to a previously described estrus cycle of slow lorises and was observable in the presence of both males as well as when the female was housed singly. Additionally, vaginal smears collected from the female close to the peak period of her vocal activity showed signs of proestrus and estrus. The acoustic properties of the whistles, specifically that they did not overlap with or were loud enough to exceed background noise commonly occurring in zoos, made them perfect candidates for analyses involving automatic processing of a large number of recordings. We conclude that bioacoustics represents a promising, completely noninvasive and relatively easily applicable tool that allows detection and anticipation of the estrus cycle in some females, thus improving the social management of slow lorises living in zoos.

Variation in Gut Microbiota of Captive Bengal Slow Lorises.

Gastrointestinal microbiome plays an important role in animal metabolism, immune system and pathology associated with health and disease. Many wild slow lorises were confiscated from illegal trade into captivities and experienced a range of changes in living environment and diet. Microbiome analysis contributes to improving captive management by identifying the alteration in their gastrointestinal microbial communities and aiding in determining the factors affecting the health of captive slow lorises. The fecal samples of eighteen Bengal slow lorises (Nycticebus bengalensis) were used to compare gut microbiota from four rescue centers located in Dehong, Gejiu, Nanning and Puer cities of China. The results showed a significant site-dependent difference in microbial community diversity. Similar to other Lorisinae species, the Phyla including Bacteroidetes, Firmicutes and Proteobacteria dominated their gut microbiome composition. The Gejiu group exhibited a higher overall diversity and the unique OTUs, which is resulted from long-term isolated husbandry and heavy human disturbances. The scarcity of gums in the captive diet was likely to cause a lower abundance of Prevotella associated with soluble fiber degradation. The variation of intestinal microbiota in different environments highlights the necessity to improve feed preparation and husbandry management for the captive Bengal slow lorises.

Artificial canopy bridges improve connectivity in fragmented landscapes: The case of Javan slow lorises in an agroforest environment.

Canopy bridges are increasingly used to reduce fragmentation in tropical habitats yet monitoring of their impact on the behavior of primates remains limited. The Javan slow loris (Nycticebus javanicus) is endemic to Java, Indonesia, where the species most often occurs in human-dominated, highly patchy landscapes. Slow lorises cannot leap, are highly arboreally adapted, and are vulnerable on the ground. To increase arboreal connectivity, as part of a long-term conservation project in Cipaganti, West Java, we built and monitored seven slow lorises bridges of two types-waterline or rubber-and monitored their use by seven adult individuals from 2016 to 2017. Motion triggered camera traps collected data for 195 ± standard deviation (SD) 85 days on each bridge. We collected 341.76 hr (179.67 hr before and 162.09 hr after the installation of bridges) of behavioral and home range data via instantaneous sampling every 5 min, and terrestrial behavior (distance and duration of time spent on the ground) via all occurrences sampling. We found that slow lorises used bridges on average 12.9 ± SD 9.7 days after their installment mainly for traveling. Slow lorises showed a trend toward an increase in their home range size (2.57 ha before, 4.11 ha after; p = 0.063) and reduced ground use (5.98 s/hr before, 0.43 s/hr; p = 0.063) after implementation of bridges. Although the number of feeding trees did not change, new feeding trees were included in the home range, and the proportion of data points spent traveling and exploring significantly decreased (p = 0.018). Waterline bridges serve a purpose to irrigate the crops of local farmers who thus help to maintain the bridges, and also ascribe value to the presence of slow lorises. Other endemic mammal species also used the bridges. We advocate the use and monitoring of artificial canopy bridges as an important supplement for habitat connectivity in conservation interventions.

Species Identification and Unlocking Hidden Genetic Diversity of Confiscated Slow Lorises (Nycticebus spp.) Based on Mitochondrial DNA Markers.

Confiscated slow lorises (Nycticebus spp.) at Bangpra Water-Bird Breeding Center (BWBC) in Thailand provided an opportunity to demonstrate the application of noninvasive genetic approaches for species identification when morphology of the animals was ambiguous. The slow lorises at BWBC had been assigned to either N. bengalensis or N. pygmaeus, based on body size. However, the morphology of N. bengalensis is highly variable and overlaps with that of N. coucang (sensu stricto). Phylogenetic analysis of cytochrome b and d-loop mitochondrial regions placed all confiscated N. pygmaeus with the published sequences of N. pygmaeus and distinguished them from other Nycticebus. All other confiscated individuals formed a monophyletic clade, most individuals grouping with published N. bengalensis sequences from wild populations in Vietnam and distinct from Peninsular Malaysian and Sumatran N. coucang, Javan N. javanicus and Bornean N. menagensis. Six individuals within the N. bengalensis clade formed a separate subgroup that did not group with any reference material as indicated by phylogenetic and haplotype network analyses. Whether these trafficked individuals are undiscovered wild populations will require further investigation. Additional genetic studies of wild slow loris populations in different regions are therefore urgently required for reference to aid the protection and conservation of these threatened species.

A Novel Method for Assessing Enamel Thickness Distribution in the Anterior Dentition as a Signal for Gouging and Other Extractive Foraging Behaviors in Gummivorous Mammals.

Gummivory poses unique challenges to the dentition as gum acquisition may often require that the anterior teeth be adapted to retain a sharp edge and to resist loading because they sometimes must penetrate a highly obdurate substrate during gum extraction by means of gouging or scraping. It has been observed previously that the enamel on the labial surface of the teeth used for extraction is thicker relative to that on the lingual surface in taxa that extract gums, while enamel is more evenly distributed in the anterior teeth of taxa that do not regularly engage in extractive behaviors. This study presents a quantitative methodology for measuring the distribution of labial versus lingual enamel thickness among primate and marsupial taxa in the context of gummivory. Computed microtomography scans of 15 specimens representing 14 taxa were analyzed. Ten measurements were taken at 20% intervals starting from the base of the crown of the extractive tooth to the tip of the cutting edge across the lingual and labial enamel. A method for including worn or broken teeth is also presented. Mann-Whitney U tests, canonical variates analysis, and between-group principal components analysis were used to examine variation in enamel thickness across taxa. Our results suggest that the differential distribution of enamel thickness in the anterior dentition can serve as a signal for gouging behavior; this methodology distinguishes between gougers, scrapers, and nonextractive gummivores. Gouging taxa are characterized by significantly thicker labial enamel relative to the lingual enamel, particularly towards the crown tip. Examination of enamel thickness patterning in these taxa permits a better understanding of the adaptations for the extraction of gums in extant taxa and offers the potential to test hypotheses concerning the dietary adaptations of fossil taxa.

Extraordinary grip strength and specialized myology in the hyper-derived hand of Perodicticus potto?

Previous behavioral reports of the African lorisid, Perodicticus potto, have speculated that these animals have an extraordinary grip strength. This ability is hypothesized to be facilitated by a range of anatomical features within the forelimb, ranging from the presence of a retia mirabilia in its wrist to the hyper-abduction of its pollex. Despite numerous behavioral reports, however, this claim of extraordinary grip strength has not been empirically substantiated. This study quantifies the physiological cross-sectional area of the digital flexor muscles within P. potto. These data are compared with a broad primate sample, including several similarly sized strepsirrhines. Contrary to expectation, we found that P. potto actually has relatively below-average digital flexor PCSA. However, we identified other myological characteristics in the upper limb of P. potto that were unexpected, including the largest brachioradialis muscle (an elbow flexor) among our primate sample, and - despite P. potto having only a vestigial second digit - an independent digital extensor indicis that is absent in almost a quarter of our primate sample.

Nasal airflow in the pygmy slow loris (Nycticebus pygmaeus) based on a combined histological, computed tomographic and computational fluid dynamics methodology.

'Macrosmatic' mammals have dedicated olfactory regions within their nasal cavity and segregated airstreams for olfaction and respiratory air-conditioning. Here, we examined the 3D distribution of olfactory surface area (SA) and nasal airflow patterns in the pygmy slow loris (Nycticebus pygmaeus), a primate with primitive nasal cavities, except for enlarged eyes that converge upon the posterodorsal nasal region. Using the head of an adult loris cadaver, we co-registered micro-computed tomography (CT) slices and histology sections to create a 3D reconstruction of the olfactory mucosa distribution. Histological sections were used to measure olfactory surface area and to annotate CT reconstructions. The loris has a complex olfactory recess (∼19% of total nasal SA) with multiple olfactory turbinals. However, the first ethmoturbinal has a rostral projection that extends far anterior to the olfactory recess, lined by ∼90% non-olfactory epithelium. Only one (of three) frontoturbinals bears olfactory mucosa. Computational fluid dynamics simulations of nasal airflow and odorant deposition revealed that there is some segregation of respiratory and olfactory flow in the loris nose, but that it is not as distinct as in well-studied 'macrosmats' (e.g. the dog). In the loris, airflow is segregated medially and laterally to vertically elongated, plate-like first ethmoturbinals. Thus, lorises may be said to have certain macrosmatic anatomical characteristics (e.g. olfactory recess), but not segregated nasal airflow patterns that are optimized for olfaction, as in canids. These results imply that a binary 'microsmatic/macrosmatic' dichotomy does not exist. Rather, mammals appear to exhibit complex trends with respect to specialization of the turbinals and recesses.

Slow loris density in a fragmented, disturbed dry forest, north-east Thailand.

Primate survival in disturbed forests can be governed by a complex of forest variables. For nocturnal arboreal primates, determining these ecological features is notoriously difficult but is critically important for their conservation. Here we assessed the effects of forest type, food availability, human disturbance, and forest structure on the nocturnal Bengal slow loris (Nycticebus bengalensis) in Sakaerat Biosphere, north-east Thailand, a small disturbed fragment containing dry evergreen, dry dipterocarp, and plantation forests. Distance sampling revealed plantation forest had the highest density estimate (27 loris/km2 ) followed by dry evergreen forest (17 loris/km2 ), while dry dipterocarp forest only had four detections. Based on forest type selection analyses, loris had a positive association with sap and fruit-flower productivity which was highest in plantation forest compared to the natural dry evergreen or dry dipterocarp forests. Furthermore, forest structure selection analysis indicated loris preferred denser canopies. Loris presence was positively associated with canopy height, canopy connections, tree trunk vegetation cover, and canopy density. The higher loris density found in the plantations, as also reported by other studies, was associated with dense vegetation structure and higher food productivity, particularly sap. However, the important structural features found in the plantation forest are strongly dependent on forest maturity, and the reduced longevity of such exotic tree species may limit the conservation importance of plantation forest, merely acting as a short-term alternative to natural habitat.

Potential Rediscovery of a Distinct Slow Loris Nycticebus (Mammalia: Strepsirrhini: Lorisidae) from Pulau Tioman.

Several slow loris (Nycticebus) sightings have occurred on the island of Pulau Tioman, Peninsular Malaysia, from 2011 to 2018. Records discussed here represent the first confirmed sightings and photographic evidence of Nycticebus on Tioman since its discovery in 1915, refuting presumptions that the Tioman slow loris is extinct. Although originally considered a subspecies of the Sunda slow loris (Nycticebus coucang), several morphological characteristics apparent in all observed individuals, including the white interocular stripe, rufous colouration and pale dorsal stripe, are similar to the Philippine slow loris (Nycticebus menagensis). Further, the broad snout and ears may be unique to this population and suggest that the population may be distinct. I, therefore, recommend that future studies consider the taxonomic status of remote and isolated Nycticebus populations given the possibility that they may represent distinct and unrecognised taxa.

The Use of Ultrasonic Communication to Maintain Social Cohesion in the Javan Slow Loris (Nycticebus javanicus).

Only a handful of primate taxa use ultrasonic vocalisations (those ≥20 kHz) to communicate. The extent and uses of ultrasonic communication remain poorly understood, potentially ranging from echolocation, advertisement of reproductive status and resource availability, social cohesion, to predator avoidance. Here, using active acoustics whereby the study subjects were observed throughout their activity period, we describe the first purely ultrasonic call from a strepsirrhine primate (family Lorisidae), recorded in a completely wild setting, and hypothesise about its function. We identified one type of ultrasonic call, the doublet click, from 14 Javan slow lorises (Nycticebus javanicus) produced by males and females of juvenile, subadult and adult ages within their social groups (n = 791, mean = 46.0 kHz). We ran quadratic discriminant function analysis, finding dominant frequency and doublet click duration as the key parameters for identifying individuals' sex and age. Significantly more vocalisations were produced during affiliative social behaviour, suggesting that the call serves a social cohesion function. Considering the range of other cryptic behaviours, including slow and silent locomotion, and the high degree of territoriality associated with venomous attacks on conspecifics, the call may also serve as a safety strategy, allowing family members to regulate distance from other slow lorises and to communicate cryptically whilst avoiding predators.

Assessing Public Perceptions and Solutions to Human-Monkey Conflict from 50 Years in Sri Lanka.

Sri Lanka is a biodiversity hotspot with high human density that contributes to increasing human-monkey conflict (HMC). In 50 years of primate studies there, the development of HMC has been documented, and many workshops and interventions organized to ameliorate HMC. These activities prompted the present survey. In the extensive lowland dry zone of Sri Lanka, the affected nonhuman primates are the toque macaque, gray and purple-faced langurs and slender loris. We surveyed and evaluated the attitudes of rural residents towards these four species in an effort to contribute to an ethnoprimatological approach to conservation, i.e., promote a coexistence and sharing of habitat between humans and monkeys. We selected 13 villages near Polonnaruwa, located centrally in the dry zone. The four nonhuman primate species differ in their behavioral ecologies, and this influenced how frequently they were thought of as pests. Most HMC was with the macaque and gray langur, less with the purple-faced langur and least with the loris. The underlying sentiment among stakeholders towards monkeys was generally either neutral or positive. Nonetheless, the majority (80%) of people desired a translocation of the troublesome monkeys from their properties to protected areas, which is impractical. Few (< 1%) openly wanted monkeys destroyed. While a traditional reverence for monkeys provides a solid basis for science and media-based education, it also contributes to the feeding of monkeys and consequent unnatural population growth, and enhanced HMC. Public understanding of the underlying causes of HMC was poor, hindering effective solutions. A combination of a feeding ban, possibly contraceptive intervention at localized HMC trouble spots, and extensive education may be the only benign alternatives to the destruction of wild primates by a powerful minority. Coexistence through strengthening and expansion of exclusive suitable protected habitats for all wildlife is a priority.

Anaphylactic shock following a bite of a wild Kayan slow loris (Nycticebus kayan) in rural Sarawak, Malaysian Borneo.

Nycticebus spp, commonly known as the slow lorus, is a small nocturnal primate found mainly in Asia. The adult slow loris weighs between 265 g and 1150 g depending on the type of species. It has a characteristic round head with large, forward-facing eyes. Slow lorises are known for their poisonous bite and are the only venomous primates. To date, there have been two published cases of slow loris bite in humans. This case report illustrates a case of anaphylactic shock following a bite of a wild Kayan slow loris (Nycticebus kayan) to a young man at Mulu District, in a remote area of Sarawak, Malaysian Borneo. The patient developed dyspnoea, a feeling of suffocation, swollen lips and cramp-like sensations over both hands. He subsequently developed syncope and hypotension. The patient was clinically stable following intramuscular injection of adrenaline 0.5 mg stat dose.

Using a multi-gene approach to infer the complicated phylogeny and evolutionary history of lorises (Order Primates: Family Lorisidae).

Extensive phylogenetic studies have found robust phylogenies are modeled by using a multi-gene approach and sampling from the majority of the taxa of interest. Yet, molecular studies focused on the lorises, a cryptic primate family, have often relied on one gene, or just mitochondrial DNA, and many were unable to include all four genera in the analyses, resulting in inconclusive phylogenies. Past phylogenetic loris studies resulted in lorises being monophyletic, paraphyletic, or an unresolvable trichotomy with the closely related galagos. The purpose of our study is to improve our understanding of loris phylogeny and evolutionary history by using a multi-gene approach. We used the mitochondrial genes cytochrome b, and cytochrome c oxidase subunit 1, along with a nuclear intron (recombination activating gene 2) and nuclear exon (the melanocortin 1 receptor). Maximum Likelihood and Bayesian phylogenetic analyses were conducted based on data from each locus, as well as on the concatenated sequences. The robust, concatenated results found lorises to be a monophyletic family (Lorisidae) (PP ≥ 0.99) with two distinct subfamilies: the African Perodictinae (PP ≥ 0.99) and the Asian Lorisinae (PP ≥ 0.99). Additionally, from these analyses all four genera were all recovered as monophyletic (PP ≥ 0.99). Some of our single-gene analyses recovered monophyly, but many had discordances, with some showing paraphyly or a deep-trichotomy. Bayesian partitioned analyses inferred the most recent common ancestors of lorises emerged ∼42 ±â€¯6 million years ago (mya), the Asian Lorisinae separated ∼30 ±â€¯9 mya, and Perodictinae arose ∼26 ±â€¯10 mya. These times fit well with known historical tectonic shifts of the area, as well as with the sparse loris fossil record. Additionally, our results agree with previous multi-gene studies on Lorisidae which found lorises to be monophyletic and arising ∼40 mya (Perelman et al., 2011; Pozzi et al., 2014). By taking a multi-gene approach, we were able to recover a well-supported, monophyletic loris phylogeny and inferred the evolutionary history of this cryptic family.