Rhinopithecus Brelichi Research Articles

Study on the microbiome and resistome of wild and captive Guizhou golden monkeys (Rhinopithecus brelichi) in China

The knowledge of gut microbial information helps evaluate the nutrition and health of the host and can be used as an important tool for improving wildlife conservation and management. Guizhou golden monkey (Rhinopithecus brelichi) is on the International Union for Conservation of Nature (IUCN 2023) Red List. The purpose of the current study was to investigate the difference of gut microbiome and resistome among three different groups of Guizhou golden monkeys using a metagenome approach, thus providing valuable insights and tools for management of captive golden monkeys. Fecal samples of Guizhou golden monkeys were collected from three places including Fanjingshan National Nature Reserve in Guizhou (n =3, Wild), Panxi wildlife rescue center in Guizhou (n=3, CapGZ), and Beijing Zoo (n=3, CapBJ). Fecal DNA was extracted and libraries were prepared and then were sequenced on Illumina PE150 platform with 2 × 150 bp paired reads. The results showed that wild Guizhou golden monkeys had lower abundance of Firmicutes, Uroviricota, Spirochaetes, Fibrobacterota, Campylobacterota, and Candidatus Melainabacteria and higher abundance Proteobacteria, Bacteroidetes and Actinobacteria than the two groups of captive monkeys. PCoA analysis showed that fecal samples from three groups could be distinctly separated. Wild monkeys had significantly higher abundance of genes associated with metabolic function, including carbohydrate metabolism, amino acid metabolism, energy metabolism, and metabolism of cofactors and vitamins. Among the top 20 ARGs analyzed, more ARGs were detected in the wild monkeys than the rest two groups. Overall, this study provides a comprehensive analysis of microbiomes and resistome in wild and captive Guizhou golden monkeys. The data is important for improving the health, management, and conservation of Guizhou golden monkeys based on the deviations in gut microbiome. It also establishes the scientific foundation for future comparative studies on the gut microbiome and resistome of Guizhou golden monkeys and other closely related Rhinopithecus species.

Population Viability and Genetic Structure of the Last Remaining Population of the Critically Endangered Gray Snub-Nosed Monkey (Rhinopithecus brelichi)

Population Viability and Genetic Structure of the Last Remaining Population of the Critically Endangered Gray Snub-Nosed Monkey (Rhinopithecus brelichi)

The Conservation Implications of the Gut Microbiome for Protecting the Critically Endangered Gray Snub-Nosed Monkey (Rhinopithecus brelichi).

The gut microbiota plays a crucial role in regulating energy metabolism, facilitating nutrient absorption, and supporting immune function, thereby assisting the host in adapting to seasonal dietary changes. Here, we compare the gut microbiome composition of wild gray snub-nosed monkeys during winter (from October to December) and spring (from January to March) to understand differences in seasonal nutrient intake patterns. Snub-nosed monkeys are foregut fermenters and consume difficult-to-digest carbohydrates and lichen. To examine the digestive adaptations of gray snub-nosed monkeys, we collected 14 fresh fecal samples for DNA analysis during the winter and spring. Based on 16S rRNA sequencing, metagenomic sequencing, and functional metagenomic analyses, we identified that Firmicutes, Actinobacteria, Verrucomicrobia, and Bacteroidetes constitute a keystone bacterial group in the gut microbiota during winter and spring and are responsible for degrading cellulose. Moreover, the transition in dietary composition from winter to spring was accompanied by changes in gut microbiota composition, demonstrating adaptive responses to varying food sources and availability. In winter, the bacterial species of the genera Streptococcus were found in higher abundance. At the functional level, these bacteria are involved in fructose and mannose metabolism and galactose metabolism c-related pathways, which facilitate the breakdown of glycogen, starch, and fiber found in fruits, seeds, and mature leaves. During spring, there was an increased abundance of bacteria species from the Prevotella and Lactobacillus genera, which aid the digestion of protein-rich buds. Combined, these findings reveal how the gut microbiota adjusts to fluctuations in energy balance and nutrient intake across different seasons in this critically endangered species. Moreover, we also identified Pseudomonas in two samples; the presence of potential pathogens within the gut could pose a risk to other troop members. Our findings highlight the necessity of a conservation plan that focuses on protecting vegetation and implementing measures to prevent disease transmission for this critically endangered species.

Analysis of winter diet in Guizhou golden monkey (Rhinopithecus brelichi) using DNA metabarcoding data.

The Guizhou golden monkey (Rhinopithecus brelichi) is a critically endangered wildlife species, and understanding its diet composition may be useful for assessing its feeding strategies. DNA metabarcoding was used to determine the dietary diversity of R. brelichi. DNA was extracted from 31 faecal samples and amplified chloroplast rbcL and mitochondrial COI DNA was sequenced using the Illumina NovaSeq platform. A comparative analysis of the sequences revealed that the five most abundant plant genera were Magnolia, Morinda, Viburnum, Tetradium and Eurya. In winter, R. brelichi mostly consumed shrubs, herbs and shrubs/trees according to the habit of plant genera with higher abundances comparatively. The five most abundant families in animal diet were Psychodidae, Trichinellidae, Staphylinidae, Scarabaeidae and Trichoceridae. This study is the first to show the composition of the winter animal diets of R. brelichi based on DNA metabarcoding. These results provide an important basis for understanding the diet of wild R. brelichi, which inhabits only the Fanjingshan National Nature Reserve, China.

Interspecific associations between <i>Rhinopithecus brelichi</i> and its sympatric species using infrared cameras

Interspecific associations between <i>Rhinopithecus brelichi</i> and its sympatric species using infrared cameras

The Food Niche Overlap and Interspecific Relationship between the Sympatric Tibetan Macaque and Grey Snub-Nosed Monkey.

Assessing the trophic niche and interspecific relationships between related species and determining how the species maintain differences in nutritional niches while coexisting in the same area are important topics in ecological research. Therefore, exploring the mechanism of food resource utilization, competition and coexistence among species distributed in the same region is important. In this study, we used fecal samples and metagenome sequencing technology to study the plant feeding habits and coexistence mechanisms of Tibetan macaques (Macaca thibetana) and grey snub-nosed monkeys (Rhinopithecus brelichi) within the same area. In the winter of 2020, we collected a total of 40 fecal samples from Tibetan macaques and grey snub-nosed monkeys; of those, 29 samples were considered valid and were analyzed using DNA metabarcoding. The results showed that in winter, Tibetan macaques consumed plants from 117 families and 184 genera, whereas grey snub-nosed monkeys consumed plants from 109 families and 165 genera. Diversity analysis revealed that there was a significant difference in the food composition of Tibetan macaques and grey snub-nosed monkeys. Tibetan macaques had a broader food niche width than grey snub-nosed monkeys at the family and genus levels. In winter, the food niches of Tibetan macaques and grey snub-nosed monkeys almost entirely overlapped (0.99). Our research provides detailed dietary data for Tibetan macaques and grey snub-nosed monkeys and valuable information that can aid in conservation efforts targeting these species.

Hybrid origin of a primate, the gray snub-nosed monkey.

Hybridization is widely recognized as promoting both species and phenotypic diversity. However, its role in mammalian evolution is rarely examined. We report historical hybridization among a group of snub-nosed monkeys (Rhinopithecus) that resulted in the origin of a hybrid species. The geographically isolated gray snub-nosed monkey Rhinopithecus brelichi shows a stable mixed genomic ancestry derived from the golden snub-nosed monkey (Rhinopithecus roxellana) and the ancestor of black-white (Rhinopithecus bieti) and black snub-nosed monkeys (Rhinopithecus strykeri). We further identified key genes derived from the parental lineages, respectively, that may have contributed to the mosaic coat coloration of R. brelichi, which likely promoted premating reproductive isolation of the hybrid from parental lineages. Our study highlights the underappreciated role of hybridization in generating species and phenotypic diversity in mammals.

Recent Advances in Genetics and Genomics of Snub-Nosed Monkeys (Rhinopithecus) and Their Implications for Phylogeny, Conservation, and Adaptation.

The snub-nosed monkey genus Rhinopithecus (Colobinae) comprises five species (Rhinopithecus roxellana, Rhinopithecus brelichi, Rhinopithecus bieti, Rhinopithecus strykeri, and Rhinopithecus avunculus). They are range-restricted species occurring only in small areas in China, Vietnam, and Myanmar. All extant species are listed as endangered or critically endangered by the International Union for Conservation of Nature (IUCN) Red List, all with decreasing populations. With the development of molecular genetics and the improvement and cost reduction in whole-genome sequencing, knowledge about evolutionary processes has improved largely in recent years. Here, we review recent major advances in snub-nosed monkey genetics and genomics and their impact on our understanding of the phylogeny, phylogeography, population genetic structure, landscape genetics, demographic history, and molecular mechanisms of adaptation to folivory and high altitudes in this primate genus. We further discuss future directions in this research field, in particular how genomic information can contribute to the conservation of snub-nosed monkeys.

Limited Heredity Diversity of the Critically Endangered Guizhou Golden Monkeys

Guizhou golden monkey (Rhinopithecus brelichi) was a unique, endangered and endemic primate species in Guizhou Province. It was an isolated population caused by habitat loss and fragmentation due to the human disturbance in the recently 30 years in China, only distributed in Fanjingshan National Natural Reserve, Guizhou Province. To know the background with demonic population structure, we sequenced 867 bp of the mitochondrial DNA D-loop from 312 fresh fecal samples, results showed there 11 haplotypes among these samples, h was 0.517, π was 0.00413. It indicated that this species had the lowest genetic diversity among four golden monkeys in China and need strengthen the conversation concern for this species immediately.

Allometric escape and acoustic signal features facilitate high-frequency communication in an endemic Chinese primate

The principle of acoustic allometry-the larger the animal, the lower its calls' fundamental frequency-is generally observed across terrestrial mammals. Moreover, according to the Acoustic Adaptation Hypothesis, open habitats favor the propagation of high-frequency calls compared to habitats with complex vegetational structures. We carried out playback experiments in which the calls of the Guizhou snub-nosed monkey (Rhinopithecus brelichi) were used as stimuli in sound attenuation and degradation experiments to test the hypothesis that propagation of Guizhou snub-nosed monkey calls is favored above vs through the forest floor vegetation. We found that low-pitched Guizhou snub-nosed monkey vocalizations suffered less attenuation than its high-pitched calls. Guizhou snub-nosed monkeys were observed emitting high-pitched calls from 1.5 to 5.0m above the ground. The use of high-pitched calls from these heights coupled with the concomitant behavior of moving about above the understory may provide a signal for receivers which maximizes potential transmission and efficacy. Our results support the Acoustic Adaptation Hypothesis and suggest that by uncoupling its vocal output from its size, this monkey can produce a high-pitched call with a broad spectral bandwidth, thereby increasing both its saliency and the frequency range over which the animal may more effectively communicate in its natural habitat.

Habitat estimates reveal that there are fewer than 400 Guizhou snub-nosed monkeys, Rhinopithecus brelichi, remaining in the wild

Population size is a critical indicator used to assess the demographic profile and conservation status of animal species living in disturbed habitats. Using GIS technology, we calculated the remaining habitat area and estimated the population size of the Guizhou snub-nosed monkey (Rhinopithecus brelichi), an Endangered primate endemic to China. The last remaining population of this species resides in the Fanjingshan National Nature Reserve (FNNR). We combined data on forest cover and patterns of group movement to estimate the size and continuity of their habitat. The results indicated that the area of remaining suitable habitat is 69.6 km2. However the monkeys are confined to an area of only 27.8 km2 (6.6% of the total reserve area of 419 km2) located in the northern part of the reserve. The remaining 28.5 km2 of suitable habitat is located in the southern part of the reserve and inaccessible to the monkeys because of the recent construction of an aerial tram. Based on estimates of population density, we calculated that the remaining world’s population of R. brelichi is approximately 125–336 individuals. In order to protect this species from extinction, we recommend that the highly fragmented evergreen and deciduous broadleaf mixed forest adjacent to the remaining troop’s home range be given protected status and an active program of forest regeneration in this area be implemented immediately.

Altitudinal ranging of the Guizhou golden monkey (Rhinopithecus brelichi): Patterns of habitat selection and habitat use

Between September 2011 and August 2013, we studied patterns of habitat selection and habitat use in the Guizhou golden monkey (Rhinopithecus brelichi) at Fanjingshan National Nature Reserve, China. the monkeys ranged across elevations between 1432 and 2100 m. Within this altitudinal range we recorded 125 genera, 72 families, and 236 tree and vine species. From these, the Guizhou golden monkey was observe to consume food items from 104 species, 51 genera, and 31 plant families. Individual food species exploited by the monkeys varied significantly across seasons and by altitude. From October to March (Spring/Winter), the monkeys foraged across their entire 700 m elevation range. From April to September, however, individuals primarily restricted their activities to a narrow zone of between 1,700 m and 1,900 m. Our data indicate that seasonal changes in the Guizhou golden monkey dietary and ranging behavior are attributable to habitat and altitudinal specific differences in the availability of plant foods. The fact that the Guizhou golden monkey actively targets common plant foods appears to represent a low energy foraging strategy designed to minimize search time and travel. Finally, due to their ranging pattern associated with habitat specificity, all of the remaining forested habitat between 1432 and 2100 m should be protected.

Diet Versus Phylogeny: a Comparison of Gut Microbiota in Captive Colobine Monkey Species

Both diet and host phylogeny shape the gut microbial community, and separating out the effects of these variables can be challenging. In this study, high-throughput sequencing was used to evaluate the impact of diet and phylogeny on the gut microbiota of nine colobine monkey species (N=64 individuals). Colobines are leaf-eating monkeys that fare poorly in captivity-often exhibiting gastrointestinal (GI) problems. This study included eight Asian colobines (Rhinopithecus brelichi, Rhinopithecus roxellana, Rhinopithecus bieti, Pygathrix nemaeus, Nasalis larvatus, Trachypithecus francoisi, Trachypithecus auratus, and Trachypithecus vetulus) and one African colobine (Colobus guereza). Monkeys were housed at five different captive institutes: Panxi Wildlife Rescue Center (Guizhou, China), Beijing Zoo, Beijing Zoo Breeding Center, Singapore Zoo, and Singapore Zoo Primate Conservation Breeding Center. Captive diets varied widely between institutions, but within an institution, all colobine monkey species were fed nearly identical or identical diets. In addition, four monkey species were present at multiple captive institutes. This allowed us to parse the effects of diet and phylogeny in these captive colobines. Gut microbial communities clustered weakly by host species and strongly by diet, and overall, colobine phylogenetic relationships were not reflected in gut microbiota analyses. Core microbiota analyses also identified several key taxa-including microbes within the Ruminococcaceae and Lachnospiraceae families-that were shared by over 90% of the monkeys in this study. Microbial species within these families include many butyrate producers that are important for GI health. These results highlight the importance of diet in captive colobines.

Genomic analysis of snub-nosed monkeys (Rhinopithecus) identifies genes and processes related to high-altitude adaptation.

The snub-nosed monkey genus Rhinopithecus includes five closely related species distributed across altitudinal gradients from 800 to 4,500 m. Rhinopithecus bieti, Rhinopithecus roxellana, and Rhinopithecus strykeri inhabit high-altitude habitats, whereas Rhinopithecus brelichi and Rhinopithecus avunculus inhabit lowland regions. We report the de novo whole-genome sequence of R. bieti and genomic sequences for the four other species. Eight shared substitutions were found in six genes related to lung function, DNA repair, and angiogenesis in the high-altitude snub-nosed monkeys. Functional assays showed that the high-altitude variant of CDT1 (Ala537Val) renders cells more resistant to UV irradiation, and the high-altitude variants of RNASE4 (Asn89Lys and Thr128Ile) confer enhanced ability to induce endothelial tube formation in vitro. Genomic scans in the R. bieti and R. roxellana populations identified signatures of selection between and within populations at genes involved in functions relevant to high-altitude adaptation. These results provide valuable insights into the adaptation to high altitude in the snub-nosed monkeys.

An efficient protocol for genomic DNA extraction from the endangered species Rhinopithecus brelichi

The Guizhou snub-nosed monkey (Rhinopithecus brelichi) is an endangered primate with approximately 750 individuals living in the Mount Fanjing area of northwestern Guizhou province. It is required to understand its evolutionary history and genetic diversity in order to take better measures for protecting this endangered species. As a widely accepted DNA source, faeces of snub-nosed monkeys were collected and used for DNA extraction. We modified the traditional method by washing the intestinal exfoliated cells with acetone and treated the cell lysate with high temperature (just below boiling point at 94 °C). We obtained a high-quality DNA, which was used for polymerase chain reactions (PCRs). Taking the mitochondrial genes (including ND6, tRNA-Glu and Cyto B region) of Sichuan snub-nosed monkey as reference, we performed a PCR and efficiently obtained DNA fragments with expected size. Further sequencing results showed that the amplified mitochondrial gene fragments had a high identity (percentage) with the published gene sequence of Sichuan snub-nosed monkey. These results demonstrated that this modified protocol is reliable and can be used in the genetic studies of monkeys and related species.

Local People’s Knowledge and Attitudes Matter for the Future Conservation of the Endangered Guizhou Snub-Nosed Monkey (Rhinopithecus brelichi) in Fanjingshan National Nature Reserve, China

Ethnoprimatology seeks to untangle the complex relationship between human and nonhuman primates, and in doing so, can provide a better understanding of how the local cultural context affects conservation initiatives. Fanjingshan National Nature Reserve in China is the last stronghold for the remaining global population of the Endangered Guizhou snub-nosed monkey (Rhinopithecus brelichi). In an effort to contribute to conservation management plans, we aimed to explore local people’s knowledge and attitudes toward the Guizhou snub-nosed monkey and conservation in the reserve using an ethnoprimatological approach. We conducted ethnographic interviews, involving structured, semistructured, and open-ended interview techniques, with 104 households in 11 villages located in and around the reserve. The results indicate that knowledge about the reserve and the monkey is unevenly distributed among respondents; men are significantly more knowledgeable about the reserve than women and women are significantly more knowledgeable about the monkey than men. Respondents are aware of the rules of the reserve but do not always agree with the rules or understand the rationale behind them. Nonetheless, respondents describe conservation as a trade-off and their attitudes toward the monkey and efforts to conserve it are generally positive and supportive. They expressed a feeling of connectedness with the Guizhou snub-nosed monkey because of its observable, humanlike behaviors; a mutual dependence on the forest; and a shared ancestry. Although our goal was to provide specific recommendations to park officials at our study site, our results also more broadly inform conservation management efforts for protected areas globally. For example, we recommend improving communication between reserve officials and local communities, appreciating the role local folklore can play in conservation, incorporating villagers’ perspectives into conservation planning, and implementing educational programs that target a wide demographic, with a particular emphasis on women.

Whole-genome sequencing of the snub-nosed monkey provides insights into folivory and evolutionary history.

Colobines are a unique group of Old World monkeys that principally eat leaves and seeds rather than fruits and insects. We report the sequencing at 146× coverage, de novo assembly and analyses of the genome of a male golden snub-nosed monkey (Rhinopithecus roxellana) and resequencing at 30× coverage of three related species (Rhinopithecus bieti, Rhinopithecus brelichi and Rhinopithecus strykeri). Comparative analyses showed that Asian colobines have an enhanced ability to derive energy from fatty acids and to degrade xenobiotics. We found evidence for functional evolution in the colobine RNASE1 gene, encoding a key secretory RNase that digests the high concentrations of bacterial RNA derived from symbiotic microflora. Demographic reconstructions indicated that the profile of ancient effective population sizes for R. roxellana more closely resembles that of giant panda rather than its congeners. These findings offer new insights into the dietary adaptations and evolutionary history of colobine primates.

A short note on extractive foraging behavior in gray snub‐nosed monkeys

Extractive foraging (EF) involves the exploitation of hidden or embedded foods, generally any food that is not visible to the naked eye. Therefore, EF is particularly important for survival in marginal habitats as it provides seasonal fallback foods in low food availability seasons. Although many studies consider primates' EF behavior and category, colobine species are usually categorized as non-extractive foragers and few studies quantitatively examine their EF behavior. In this study, we examined the EF behavior of one colobine species, the gray snub-nosed monkey (Rhinopithecus brelichi), at Yangaoping in Fanjingshan National Nature Reserve, Guizhou. We recorded 6 categories of EF behaviors. The most frequently sought-out foods were seeds, young bamboo and invertebrates. Extracted foods accounted for an average of 26.02% of feeding records. As the monkey engages in little EF behavior in the winter when the food availability is low, these results seem to do not support the hypothesis that EF serves to secure additional resources during lean times in marginal or seasonal habitats. According to these findings, we suggest R. brelich should be considered as an extractive forager. Our study also highlights the need for increased representation of colobines in the EF literature to better inform the discussion concerning its link to primate brain evolution.

Genetic Diversity in Endangered Guizhou Snub-Nosed Monkeys (Rhinopithecus brelichi): Contrasting Results from Microsatellite and Mitochondrial DNA Data

To evaluate the conservation status of a species or population it is necessary to gain insight into its ecological requirements, reproduction, genetic population structure, and overall genetic diversity. In our study we examined the genetic diversity of Rhinopithecus brelichi by analyzing microsatellite data and compared them with already existing data derived from mitochondrial DNA, which revealed that R. brelichi exhibits the lowest mitochondrial diversity of all so far studied Rhinopithecus species. In contrast, the genetic diversity of nuclear DNA is high and comparable to other Rhinopithecus species, i.e. the examined microsatellite loci are similarly highly polymorphic as in other species of the genus. An explanation for these differences in mitochondrial and nuclear genetic diversity could be a male biased dispersal. Females most likely stay within their natal band and males migrate between bands, thus mitochondrial DNA will not be exchanged between bands but nuclear DNA via males. A Bayesian Skyline Plot based on mitochondrial DNA sequences shows a strong decrease of the female effective population size (Nef) starting about 3,500 to 4,000 years ago, which concurs with the increasing human population in the area and respective expansion of agriculture. Given that we found no indication for a loss of nuclear DNA diversity in R. brelichi it seems that this factor does not represent the most prominent conservation threat for the long-term survival of the species. Conservation efforts should therefore focus more on immediate threats such as development of tourism and habitat destruction.

Into the night: camera traps reveal nocturnal activity in a presumptive diurnal primate, Rhinopithecus brelichi

Most living primates exhibit a daytime or nighttime activity pattern. Strict diurnality is thought to be the rule among anthropoids except for owl monkeys. Here we report the diel activity pattern of an Asian colobine, the Guizhou snub-nosed monkey Rhinopithecus brelichi, based on a methodology that relied on using 24-h continuously operating camera traps. We conducted the study in Fanjingshan National Nature Reserve in Guizhou, China from March 22 to May 19 and from June 17 to October 14, 2011. After standardizing all time elements to a meridian-based time according to the geographic coordinates of the study site, we showed unequivocally that the monkeys, though predominantly diurnal, exhibited activity beyond daylight hours throughout the study. Specifically, their activity at night and during twilight periods suggests a complex interplay of behavioral adaptations, among others, to living in a temperate environment where day length and food resources fluctuate substantially across seasons. We contend that, under prevailing ecological conditions, so-called strictly diurnal primates may adjust their activity schedule opportunistically in order to increase energy intake. We also discuss the advantages of using camera traps in primate studies, and how the standardized use of meridian-based time by researchers would benefit comparisons of diel activity patterns among primates.