Evaluation of Gut Microbiota Stability and Flexibility as a Response to Seasonal Variation in the Wild François' Langurs (Trachypithecus francoisi) in Limestone Forest.

The coevolution between gut microbiota and the host markedly influences the digestive strategies of animals to cope with changes in food sources. We have explored the compositional structure and seasonal variation in the gut microbiota of François' langur in a limestone forest in Guangxi, southwest China, using 16S rRNA sequencing. Our results demonstrated that Firmicutes and Bacteroidetes were the dominant phyla in langurs, followed by Oscillospiraceae, Christensenellaceae, and Lachnospiraceae at the family level. The top five dominant phyla did not show significant seasonal variations, and only 21 bacterial taxa differed at the family level, indicating stability in gut the microbiota possibly with respect to foraging for several dominant plants and high-leaf feeding by the langurs. Moreover, rainfall and minimum humidity are important factors affecting the gut microbiota of the langurs, but they explain few changes in bacterial taxa. The activity budget and thyroid hormone levels of the langurs did not differ significantly between seasons, indicating that these langurs did not respond to seasonal changes in food by regulating behavior or reducing metabolism. The present study indicates that the gut microbiota's structure is related to digestion and energy absorption of these langurs, providing new perspectives on their adaptation to limestone forests. IMPORTANCE François' langur is a primate that particularly lives in karst regions. The adaptation of wild animals to karst habitats has been a hot topic in behavioral ecology and conservation biology. In this study, gut microbiota, behavior, and thyroid hormone data were integrated to understand the interaction of the langurs and limestone forests from the physiological response, providing basic data for assessing the adaptation of the langurs to the habitats. The responses of the langurs to environmental changes were explored from the seasonal variations in gut microbiota, which would help to further understand the adaptive strategies of species to environmental changes.

Diet and feeding behavior of a group of high-altitude rhesus macaques: high adaptation to food shortages and seasonal fluctuations.

Diet and feeding behavior data are crucial to a deep understanding of the behavioral response and adaptation of primates to a high-altitude environment. From August 2019 to June 2021, we collected data on the feeding behavior of a high-altitude rhesus macaque Macaca mulatta group from Yajiang County, Western Sichuan Plateau, which has an altitude of over 3,500 m. The results showed that feeding (33.0 ± 1.8%) and moving (28.3 ± 2.6%) were the dominant behavior of rhesus macaques. Macaques ate 193 food items, comprising 11 food categories from 90 species. Our study found that plant roots (30.9 ± 30.1%) and young leaves (28.0 ± 33.1%) were the main foods eaten by macaques. The preferred foods of rhesus macaques were young leaves, fruits, and seeds, and the consumption of these items was positively correlated with its food availability. When the availability of preferred foods was low, macaques took plant roots, barks, and fallen leaves as fallback foods. In particular, roots were a dominant food item in winter, and this way of feeding became a key survival strategy. Our results suggest that, facing the relative scarcity and strong seasonal fluctuations of food resources in high-altitude habitat, macaques adopt active foraging strategies, relying on a variety of food species and adjusting flexibly their food choices based on food availability, which may help to maximize the energy efficiency of high-altitude macaques.

Captivity restructures the gut microbiota of François' langurs (Trachypithecus francoisi).

Gut microbiota is crucial to primate survival. Data on the gut microbiota of captive and wild animals can provide a physiological and ecological basis for the conservation of rare and endangered species. To study the effect of captivity on the gut microbiota, we examine the difference in the gut microbiota composition between captive and wild Francois' langurs (Trachypithecus francoisi), using 16S rRNA sequencing technology. The results showed that the composition of the gut microbiota of captive and wild langurs was characterized by Firmicutes (51.93 ± 10.07% vs. 76.15 ± 8.37%) and Bacteroidetes (32.43 ± 10.00% vs. 4.82 ± 1.41%) at the phylum level and was characterized by Oscillospiraceae (15.80 ± 5.19% vs. 30.21 ± 4.87%) at the family level. The alpha diversity of gut microbiota in captive langurs was higher than those in wild, such as the Shannon index (4.45 ± 0.33 vs. 3.98 ± 0.19, P < 0.001) and invSimpson index (35.11 ± 15.63 vs. 19.02 ± 4.87, P < 0.001). Principal coordinates analysis (PCoA) results showed significant differences in the composition of gut microbiota between captive and wild langurs at both the phylum and family levels (weight UniFrac algorithm, phylum level: R2 = 0.748, P = 0.001; family level: R2 = 0.685, P = 0.001). The relative abundance of Firmicutes (51.93 ± 10.07%) in captive langurs was lower than that of wild langurs (76.15 ± 8.37%), and the relative abundance of Bacteroidetes (32.43 ± 10.00%) in captive langurs was higher than that of wild (4.82 ± 1.41%). Our study concludes that dietary composition could be a crucial determinant in shaping the gut microbiota of langurs because more fiber-rich foods used by the wild langurs could increase the abundance of Firmicutes, and more simple carbohydrate-rich foods consumed by the captive langurs increase the abundance of Bacteroidetes. We highlight the importance of captivity on the gut microbiota and the need to consider the gut microbiota in animal provision.

Gut microbiota of white-headed black langurs (Trachypithecus leucocephalus) in responses to habitat fragmentation.

The white-headed black langur (Trachypithecus leucocephalus) is exclusively distributed in the karst forests and is critically endangered owing to habitat fragmentation. Gut microbiota can provide physiological data for a comprehensive study of the langur's response to human disturbance in the limestone forest; to date, data on spatial variations in the langurs' gut microbiota are limited. In this study, we examined intersite variations in the gut microbiota of white-headed black langurs in the Guangxi Chongzuo White-headed Langur National Nature Reserve, China. Our results showed that langurs in the Bapen area with a better habitat had higher gut microbiota diversity. In the Bapen group, the Bacteroidetes (13.65% ± 9.73% vs. 4.75% ± 4.70%) and its representative family, Prevotellaceae, were significantly enriched. In the Banli group, higher relative abundance of Firmicutes (86.30% ± 8.60% vs. 78.85% ± 10.35%) than the Bapen group was observed. Oscillospiraceae (16.93% ± 5.39% vs. 16.13% ± 3.16%), Christensenellaceae (15.80% ± 4.59% vs. 11.61% ± 3.60%), and norank_o__Clostridia_UCG-014 (17.43% ± 6.64% vs. 9.78% ± 3.83%) were increased in comparison with the Bapen group. These intersite variations in microbiota diversity and composition could be accounted for by differences in food resources caused by fragmentation. Furthermore, compared with the Banli group, the community assembly of gut microbiota in the Bapen group was influenced by more deterministic factors and had a higher migration rate, but the difference between the two groups was not significant. This might be attributed to the serious fragmentation of the habitats for both groups. Our findings highlight the importance of gut microbiota response for the integrity of wildlife habitats and the need in using physiological indicators to study the mechanisms by which wildlife responds to human disturbances or ecological variations.

Seasonal variations in gut microbiota of semiprovisioned rhesus macaques (Macaca mulatta) living in a limestone forest of Guangxi, China.

Assessment of gut microbiota, used to explore ecological adaptation strategies and evolutionary potential of species, provides a new viewpoint to the conservation and management of endangered animals. In this research, the gut microbiota of a group of semiprovisioned rhesus macaques (Macaca mulatta) living in a limestone forest exhibiting seasonal changes in plant items were studied to investigate the adaptation strategies of these macaques to this specific habitat. The findings revealed significant seasonal changes in the diversity and composition of the rhesus macaques' gut microbiota, which were higher in the rainy season than in the dry season. In the rainy season, Bacteroidetes (31.83 ± 16.14% vs. 19.91 ± 18.20%) were significantly increased and Prevotella (23.70 ± 15.33% vs. 15.40 ± 16.10%), UCG-002 (4.48 ± 3.16% vs. 2.18 ± 2.01%), and UCG-005 (4.22 ± 2.90% vs. 2.03 ± 1.82%) were more enriched at the genus level. In the dry season, Firmicutes significantly increased (71.84 ± 19.28% vs. 60.91 ± 16.77%), and Clostridium_sensu_stricto_1 (8.45 ± 9.72% vs. 4.76 ± 6.64%), Enterococcus (10.17 ± 13.47% vs. 0.69 ± 2.36%), and Sarcina (4.72 ± 7.66% vs. 2.45 ± 4.71%) were more enriched at the genus level. These differences in gut microbiota may be due to seasonal variations in plant items in these habitats alongside changes in the provisioned foods from tourists. Additionally, deterministic processes predominate the assembly of the macaque's gut microbiota community. This indicates that the animal's high reliance on natural plants and provisioned foods increased the impact of deterministic processes. This study concludes that a balance between provisioned foods and natural plants might be vital in shaping the gut microbiota in the macaques. Furthermore, the dynamic adjustment in gut microbiota might be a physiological mechanism for the macaques in response to the seasonal variations in the ecological factors and food provision.

Habitat use of the white-headed langurs in limestone forest of Southwest Guangxi, China: Seasonality and group size effects.

Understanding how animals cope with habitat-specific environmental factors can assist in species conservation management. We studied the habitat use of four groups (two large and two small groups) of white-headed langurs (Trachypithecus leucocephalus) living in the forest of southwest Guangxi, China between September 2016 and February 2017 via instantaneous scan sampling. Our results showed that the langurs primarily used hillsides (55.91% ± 6.47%), followed by cliffs (29.70% ± 5.48%), hilltops (7.26% ± 3.55%), flat zones (6.99% ± 6.58%), and farmlands (0.14% ± 0.28%). The langurs moved most frequently on hillsides (49.35% ± 6.97%) and cliffs (35.60% ± 9.17%). The hillsides were more frequently used (66.94% ± 7.86%) during feeding, and the langurs increased the use of hilltops during the rainy season, and the use of cliffs in the dry season. The langurs frequently rested on hillsides (49.75% ± 8.16%) and cliffs (38.93% ± 8.02%). The larger langur group used cliffs more frequently when moving and resting, whereas the small langur group used hillsides more frequently while resting. Langurs in all groups avoided the flat zones for feeding. Their use of habitat reflected the balancing of foraging needs, thermoregulation, and predator avoidance. We conclude that the ecological factors are determinants of habitat use for white-headed langurs. Our findings suggest that conservation efforts should focus on protecting the vegetation on the hillsides and restoring the vegetation on the flat zones.

Food availability, temperature, and day length drive seasonal variations in the positional behavior of white-headed langurs in the limestone forests of Southwest Guangxi, China.

Information on positional behavior contributes to the understanding of the ecological adaptation mechanisms in animals. We collected data on the positional behavior of white-headed langurs (Trachypithecus leucocephalus) at the Guangxi Chongzuo White-Headed Langur National Nature Reserve from September 2016 to August 2017 via instantaneous scan sampling method. This study aimed to examine the importance of positional behavior flexibility in limestone forests characterized by seasonal variations in climate and food resources. Our results indicated that langurs adopted leaping (47.92% ± 5.50%) and vertical climbing (40.13% ± 6.20%) as their predominant locomotor modes and sitting (83.08% ± 4.70%) as their predominant posture. Their positional behavior exhibited marked seasonal variations. More specifically, langurs used quadrupedal walking more frequently during the dry season than during the rainy months. In the stationary state, they sat more frequently during the dry season, whereas they laid and suspended more often during the rainy season. Their positional behavior was affected by fruit availability, day length, and temperature. Quadrupedal walking increased with the decrease in fruit availability, whereas leaping was positively correlated with fruit availability. Moreover, sitting was positively correlated with average temperature but negatively correlated with day length. Lying was also negatively correlated with temperature but positively correlated with day length. We conclude that white-headed langurs adapt to limestone forests with positional behavior flexibility in response to seasonality. Our research provides evidence of the effects of food availability, ambient temperature, and day length on the positional behavior of white-headed langurs, highlighting the need to understand their behavioral ecology and the influence of ecological factors on behavioral adaptation.

Interannual variation in food choice of white-headed langur inhabiting limestone forests in Fusui, southwest Guangxi, China.

Food habits are important factors in the adaptation of wild nonhuman primates. White-headed langurs (Trachypithecus leucocephalus) are endemic to heavily fragmented limestone forests and adapt to unique living environments via flexible food selection strategies. In this study, we compared the dietary data for white-headed langurs living in Chongzuo White-headed Langur National Nature Reserve in 2013 and 2016 to evaluate interannual variations in diet. Our results indicated that young leaves were the main food source for langurs, accounting for 52.4% (SD 25.4%) and 65.2% (SD 22.4%) of their diet in 2013 and 2016, respectively. The pattern of plant part consumption was similar between the two years. The consumption of young leaves varied with the availability of young leaves, whereas the consumption of mature leaves was negatively correlated with young leaf availability. The consumption of plant species and diet diversity were higher in 2013 than in 2016. In both 2013 and 2016, although diet diversity varied with the consumption of mature leaves, it was negatively correlated with the consumption and availability of young leaves. Dietary interannual variation is likely to either be linked to phenological variations or indicate that white-headed langurs have a flexible ecological adaptation coping with habitat fragmentation.

Gut microbiota are associated with sex and age of host: Evidence from semi-provisioned rhesus macaques in southwest Guangxi, China.

Host characteristics, such as sex and age, are closely associated with the structure and function of gut microbiota; however, less is known about the effects of age and sex on the gut microbiota of nonhuman primates, and therefore, our knowledge of interindividual variability in host gut microbiota is limited. In this study, 153 fecal samples from rhesus macaques (Macaca mulatta) were analyzed using high-throughput 16S rRNA sequencing in order to explore associations between age and sex of the host and their gut microbiota. The results indicated that female macaques had higher alpha diversity and a more unique gut microbiota than did males. The proportion of Proteobacteria, Tenericutes, Cyanobacteria, unclassified bacteria, and Verrucomicrobia was higher in females than that in males. We also found that adults of both sexes had a higher alpha diversity, a higher proportion of norank Ruminococcaceae, Oscillospira, norank Lachnospiraceae, norank Clostridiales, and Succinivibrio, and a lower proportion of Enterococcus than immatures. Functional analyses revealed that the richness of metabolic pathways was higher in females than males and in adults compared with immatures. These results could be attributed to differences in the nutritional requirements and hormone levels of macaques of different sex and age classes. We conclude that variation in the gut microbiota of different sex and age classes of rhesus macaques may be linked to age- and sex-specific differences in nutrient requirements and hormone levels. These results highlight the importance of host age and sex on the structure and function of the gut microbiota and the need to consider physiological traits when conducting studies on the gut microbiota.

Diet, food availability, and climatic factors drive ranging behavior in white-headed langurs in the limestone forests of Guangxi, southwest China.

Changes in abiotic and biotic factors can affect the efficiency of biological systems in animals, forcing them to adjust their behaviors in response to daily and seasonal variations. From September 2016 to August 2017, we collected ranging behavior data on four groups of white-headed langurs ( Trachypithecus leucocephalus) in the Guangxi Chongzuo White-Headed Langur National Nature Reserve, Guangxi, southwest China. We simultaneously analyzed how multiple ecological factors affect langur ranging behavior, which should facilitate our understanding of the potential mechanisms underlying their adaptation to limestone habitats. Results showed that langur ranging behavior was significantly affected by diet composition, food availability, and climatic factors. Specifically, moving time and daily path length increased with the increase in dietary diversity. Furthermore, moving time and daily path length were positively associated with the availability of fruit and relative humidity of the forest, and moderately associated with temperature and relative humidity of bare rock. Our study demonstrated that langurs maintain stable moving and feeding times and exhibit a short daily travel distance, likely adopting an energy-conserving behavioral strategy in response to food shortages and high temperatures in the fragmented karst forest. These results highlight the importance of food availability and temperature in shaping the ranging behavior of these karst-dwelling primates.

Seasonal variation in the gut microbiota of rhesus macaques inhabiting limestone forests of southwest Guangxi, China.

Data on the gut microbiota of animals can provide new insights into dietary ecology of hosts, consequently assisting in understanding their adaptation strategy and evolutionary potential. We studied the gut microbiota composition and function of the wild rhesus macaques (Macaca mulatta) using 16S rRNA sequencing method. Our results revealed that the gut microbiota of the wild rhesus macaques was dominated by Firmicutes, Bacteroidetes, and Spirochaetes. Diversity and richness of gut microbiota were higher during the dry season than the rainy season. Specifically, higher proportions of Firmicutes, Tenericutes, Cyanobacteria, and unclassified bacteria at the phylum level and more Coprococcus at the genus level were detected in the dry season. Predictive functional analysis showed that pathways associated with carbohydrate metabolism and drug resistance (antimicrobial and antineoplastic) were richer in the dry season. These seasonal differences in microbiota could be due to their heavier dependence on leaf-based diet in the dry season. Additionally, macaques in limestone forests had a higher percentage of Spirochaetes, probably suggesting that the proportion of fruits in dietary composition also play an important role in the gut microbiota. We concluded that diet was strongly linked to the diversity, composition, and function of the gut microbiota in the wild groups of rhesus macaques living in the limestone forest, highlighting the importance of diet in the gut microbiota of macaques and the need to conduct further study on the adaptation strategy in response of environmental changes in the ground of gut microbiota.

Nutrient contents predict the bamboo-leaf-based diet of Assamese macaques living in limestone forests of southwest Guangxi, China.

Determining the nutrient factors influencing food choice provides important insight into the feeding strategy of animals, which is crucial for understanding their behavioral response to environmental changes. A bamboo-leaf-based diet is rare among mammals. Animals' food choice and nutritional goals have been explained by several frameworks; however, the influence of nutrients on food choice in bamboo-leaf-based macaques is not yet available. Assamese macaques (Macaca assamensis) inhabiting limestone forests are characterized by such a bamboo-leaf-based diet, predominantly consuming young leaves of Bonia saxatilis, a shrubby, karst-endemic bamboo. We studied the feeding behavior of one group of Assamese macaques using instantaneous scan sampling in limestone forests of the Guangxi Nonggang National Nature Reserve in southwest Guangxi, China. We compared the nutrient content of staple food and nonfood items and examine the role of key nutrients in the food selection of macaques. Our results showed that young leaves of bamboo B. saxatilis contained more water, crude protein, phosphorus, and less tannin than nonfood items. Furthermore, staple foods contained a higher content of water and less content of calcium than nonfood items. More specifically, quantities of water, crude protein, calcium, and phosphorus in food items were critical factors affecting feeding time on a specific plant item. Our results suggest that young bamboo leaves could meet macaques' required protein and water intake, while enabling them to maintain their mineral balance, consequently facilitating to maintain the primates' bamboo-leaf-diet in the limestone forest. Our findings confirm the effects of nutrient contents in food choice of Assamese macaques, highlighting the importance of the nutrient contents in maintaining their bamboo-based diet and the need to increase the knowledge on their nutritional strategy adapted to the bamboo-dominated diet inhabiting the unique limestone habitat.

Behavioral adjustments and support use of François' langur in limestone habitat in Fusui, China: Implications for behavioral thermoregulation.

Climatic factors such as temperature and humidity vary seasonally in primate habitats; thus, behavioral adjustments and microhabitat selection by primate species have been interpreted as behavioral adaptations. François' langur (Trachypithecus francoisi), a native species to southwest China and northern Vietnam, inhabits a limestone habitat with extreme climatic conditions. To understand the potential effects of climatic seasonality on this species, we collected data on the individual behavioral budgets in a T. francoisi group between January and December 2010 in Fusui County, China. Monthly, we performed 5-11 days of observation during this period, using focal animal sampling and continuous recording methods. We also recorded ambient temperature (T a) and relative humidity (H r) data at our study site. Results indicated that T a and H r were significantly correlated with each other and fluctuated dramatically on a daily, monthly, and seasonal basis. The amount of time spent resting, grooming, basking, and huddling also varied on a daily, monthly, and seasonal basis. The proportion of resting time and total sedentary activity time significantly increased at high and low T as, respectively. The total sedentary time, resting time, and plant branch use all showed positive significant correlations with T a. Our results suggest that behavioral adjustment and support use of T. francoisi, at least partly, were related to thermoregulation. T. francoisi minimized thermal stress through behavioral adjustments and support use. It is an adaptive behavior associated with the climatic extremes of limestone habitat. This study can potentially advise conservation management strategies in this specific habitat. Conservation efforts should focus on vegetation restoration in langurs' habitat, including those in the foothills.

Seasonal Variation in Activity Budget of Assamese Macaques in Limestone Forest of Southwest Guangxi, China.

The activity budget is important for understanding behavioural variability and adaptation in primates. Assamese macaques (Macaca assamensis) found in the limestone forest of Guangxi Nonggang National Nature Reserve, Southwest China, primarily feed on young leaves of Bonia saxatilis (a shrubby, karst-endemic bamboo). To understand how a specific bamboo leaf-based diet and ecological factors affect activity budget, one group of Assamese macaques was studied using instantaneous scan sampling for 1 year. The macaques spent most of their time feeding (32.7 ± 5.4%), followed by resting (28.6 ± 6.3%), moving (28.6 ± 5.3%), grooming (8.0 ± 3.0%), playing (1.7 ± 1.6%) and other activities (0.4 ± 0.2%). Their activity budget was similar to that of typical frugivorous primates and bamboo-dominated primates, which spent more time on active behaviours (feeding and moving, 61.3 ± 6.0%) than on inactive behaviours (resting and grooming, 36.6 ± 6.4%). The macaques spent significantly more time resting during the fruit-lean season and more time moving during the fruit-rich season. Their activity budget was significantly affected by diet. Resting time increased with decreased fruit consumption, whereas moving time decreased with the increasing mature leaf consumption. Playing time decreased when the macaques consumed more young bamboo leaves. The activity budget was also influenced by food availability and climatic factors. Resting time increased with decreasing temperature, whereas moving time increased with increasing day length and young leaf availability. Grooming time increased with decreasing day length and increasing temperature, and playing time increased with increasing day length. Our findings provide evidence of the importance of diet, food availability, temperature and day length in coping with seasonal variation in ecological factors, highlighting the need to increase knowledge of the behavioural ecology of the Assamese macaques living in the unique limestone forest and to understand the influence of a bamboo-dominated diet and ecological factors on their survival.

Gut microbiota of provisioned and wild rhesus macaques (Macaca mulatta) living in a limestone forest in southwest Guangxi, China.

The gut microbiota plays an important role in animal health and is strongly affected by the environment. Captivity and human source food have been shown to influence drastically the gut microbiota composition and function of wild animals. Therefore, in the present study, the gut microbiota of provisioned and wild populations of limestone-living rhesus macaques (Macaca mulatta) were compared using high-throughput 16S rRNA sequencing and bioinformatic analyses. The results indicated that provisioned macaques had a higher microbial richness than wild macaques, but there was no significant difference in the evenness of the gut microbiota between the two populations. Provisioned macaques also showed a higher abundance of Firmicutes and a lower abundance of Bacteroidetes than wild macaques. Functional analysis revealed that wild macaques had enriched microbial pathways involved in glycan biosynthesis and metabolism, transport and catabolism, and the digestive and endocrine systems, while provisioned macaques were richer in pathways associated with signaling molecules and interaction, neurodegenerative diseases. These differences were likely due to modification of the gut microbiota of the provisioned macaques to enable the digestion of new foods.

Effect of Group Size on Time Budgets and Ranging Behavior of White-Headed Langurs in Limestone Forest, Southwest China.

The ecological constraints model is well supported by data from most frugivorous primates; however, the prediction power of the model is weak for folivorous primates. From September 2016 to August 2017, we collected comparative data on time budgets, daily path lengths and diets of four groups of white-headed langurs (Trachypithecus leucocephalus), including two large groups (G-DS and G-ZWY) and two small groups (G-LZ and G-NN) in Chongzuo White-Headed Langur National Nature Reserve, Guangxi Province, Southwest China. The aim was to obtain evidence of foraging competition and to test the ecological constraints model on this highly folivorous primate in its karst habitat. The results showed that langurs in the larger groups spent more time traveling, less time resting, and had a longer average daily path length than those in the small groups. Diet composition and dietary diversity were not significantly different between the large and small groups. Our study demonstrates that langurs from large groups suffer scramble competition in limestone forests and supports the validity of the ecological constraints model for folivores.

Positional Behaviours of François' Langur (Trachypithecus francoisi) in the Limestone Forest of Nonggang, Guangxi, South-West China.

Primates' positional behaviours provide information for understanding relationships among morphology, ecology and behavioural flexibility. From September 2005 to August 2006, we collected data via instantaneous scan sampling on positional behaviours in François' langurs at Nonggang Nature Reserve, south-west China. We predicted that these langurs would use (1) leaping as the predominant locomotor mode and (2) bipedal standing more frequently in the rainy season than in the dry season. Our result showed that leaping was the dominant locomotor mode (38.38%), followed by -quadrupedal walking (31.2%), vertical climbing (25.1%) and quadrupedal running (5.3%). The ground was the most frequently used stratum during movement (33.4%). Most locomotion through trees occurred on small- (48.7%) and medium-sized (47.6%) substrates. Locomotor mode, forest stratum use and substrate use during movement did not vary seasonally. When stationary, sitting was the most common posture (92.1%), followed by bipedal standing (3.7%), lying (3.5%), quadrupedal standing (0.6%), suspending (0.2%) and back-lying (<0.1%). Posture varied significantly with the season. During resting, langurs used sitting and bipedal standing more frequently in the dry season, while adopting lying more frequently in the rainy season. During feeding, sitting was adopted more frequently in the rainy than in the dry season, whereas bipedal standing was used more frequently in the dry season. Langurs spent more feeding time on the ground in the dry than in the rainy season. Locomotor patterns in François' langurs are likely linked to morphological and anatomical characteristics, along with the limestone forest's structure. Our result completely supported prediction 1 but not prediction 2. This study suggests that seasonal variation in positional behaviour might result from the temporal difference in spatial distribution of foods and behavioural thermoregulation strategy. We found that François' langurs adjusted positional behaviour in response to seasonality, and this behavioural flexibility allows them to survive in a variety of habitats, including limestone forests.

Surface-Enhanced Raman Spectroscopy on Self-Assembled Au Nanoparticles Arrays for Pesticides Residues Multiplex Detection under Complex Environment.

The high reproducibility of trace detection in complex systems is very hard but crucial to analytical technology and science. Here, we present a surface-enhanced Raman scattering (SERS) platform made by large-scale self-assembly of Au nanoparticle (NP) arrays at the cyclohexane/water interface and its use for pesticides residues trace detection. The analyte molecules spontaneously localize into the Au NPs' nanogaps during the self-assembly process, yielding excellent Raman signal enhancement by surface effects, and possibly both by the concentration of the analytes into the array and by plasmonic hot-spot formation. Transmission electron microscopy (TEM) images demonstrate a good uniformity of interparticle distances (2⁻3 nm) in the Au NP arrays. SERS experiments on crystal violet (CV) molecules demonstrated that the relative standard deviations (RSD) of the band intensities at 1173, 1376, and 1618 cm-1 were 6.3%, 6.4%, and 6.9%, respectively, indicating high reproducibility of the substrate. Furthermore, we demonstrate that two pesticides dissolved in organic and aqueous phases could be simultaneously detected, suggesting an excellent selectivity and universality of this method for multiplex detection. Our SERS platform opens vast possibilities for repeatability and sensitivity detection of targets in various complex fields.

Variations in diet composition of sympatric Trachypithecus francoisi and Macaca assamensis in the limestone habitats of Nonggang, China.

Comparative studies of sympatric species are essential for understanding behavioral and ecological adaptation as well as the mechanisms that can reduce resource competition to allow coexistence. François' langurs (Trachypithecus francoisi) and Assamese macaques (Macaca assamensis) are sympatric primate species found in the limestone seasonal rainforests of Nonggang Nature Reserve, southwestern Guangxi, China. To explore their different adaptation strategies, we collected data on diet using scan sampling at 15-min intervals. Our results revealed that François' langurs showed a more flexible diet composition than Assamese macaques. François' langurs increased dietary diversity and mature leaf consumption in response to seasonal scarcity of preferred young leaves and fruits, whereas Assamese macaques relied heavily on young bamboo leaves (Indocalamus calcicolus) in most months. These variations reflect the differences in digestive physiology, morphology, and the temporal and spatial distribution of food resources.

Effect of habitat fragmentation on ranging behavior of white-headed langurs in limestone forests in Southwest China.

The critically endangered white-headed langur (Trachypithecus leucocephalus) is confined to fragmented karst forests of southwest Guangxi Province, China. A lack of information on the influence of habitat fragmentation on langur behavior has prevented a comprehensive understanding of their ranging behavior and the development of effective langur conservation strategies. We collected comparative data on time budgets, daily path lengths, home range and diets of four langur groups inhabiting the lightly fragmented Fusui forest (G1, G2) and the more heavily fragmented Chongzuo forest (G3, G4). The aim was to explore the effect of this fragmentation on langur ranging behavior. Our results showed that the Fusui groups spent more time on moving and less time on feeding and playing than the Chongzuo groups. Daily path lengths were 472.4-536.1 m for the Fusui groups and 449.6-480.7 m for the Chongzuo groups, indicating no marked inter-site variation. The Fusui groups occupied much larger home ranges (23.8-33.8 ha) than the Chongzuo groups (14.5-15.8 ha). However, all groups had similar monthly home ranges. Diets significantly differed among langur groups. The Fusui groups consumed more young leaves and had much lower diet diversity compared with the Chongzuo groups. Our findings indicate that habitat fragmentation is one of the crucial determinants of white-headed langur ranging behavior because fragmentation reduces and restricts the home range. Langurs in fragmented habitat adopt an energy conservation strategy characterized by devoting more time to feeding and less time to moving, with a smaller home range and consumption of more plant species. We argue that linking fragmented forests with corridors should be considered a priority in a wider and comprehensive longer term langur population conservation and habitat management strategy.