Zoonotic pathogens in wild Asian primates: a systematic review highlighting research gaps.

Introduction: Ongoing global changes, including natural land conversion for agriculture and urbanization, modify the dynamics of human-primate contacts, resulting in increased zoonotic risks. Although Asia shelters high primate diversity and experiences rapid expansion of human-primate contact zones, there remains little documentation regarding zoonotic surveillance in the primates of this region. Methods: Using the PRISMA guidelines, we conducted a systematic review to compile an inventory of zoonotic pathogens detected in wild Asian primates, while highlighting the coverage of primate species, countries, and pathogen groups surveyed, as well as the diagnostic methods used across the studies. Moreover, we compared the species richness of pathogens harbored by primates across diverse types of habitats classified according to their degree of anthropization (i.e., urban vs. rural vs. forest habitats). Results and discussion: Searches of Scopus, PubMed, and the Global Mammal Parasite Database yielded 152 articles on 39 primate species. We inventoried 183 pathogens, including 63 helminthic gastrointestinal parasites, two blood-borne parasites, 42 protozoa, 45 viruses, 30 bacteria, and one fungus. Considering each study as a sample, species accumulation curves revealed no significant differences in specific richness between habitat types for any of the pathogen groups analyzed. This is likely due to the insufficient sampling effort (i.e., a limited number of studies), which prevents drawing conclusive findings. This systematic review identified several publication biases, particularly the uneven representation of host species and pathogen groups studied, as well as a lack of use of generic diagnostic methods. Addressing these gaps necessitates a multidisciplinary strategy framed in a One Health approach, which may facilitate a broader inventory of pathogens and ultimately limit the risk of cross-species transmission at the human-primate interface. Strengthening the zoonotic surveillance in primates of this region could be realized notably through the application of more comprehensive diagnostic techniques such as broad-spectrum analyses without a priori selection.

Linking glucocorticoid variations to monthly and daily behavior in a wild endangered neotropical primate.

Identifying the factors swaying physiological stress levels in wild animals can help depict how they cope with environmental and social stressors, shedding light on their feeding ecology, behavioral plasticity, and adaptability. Here, we used noninvasive methods to explore the link between glucocorticoid levels and behavior in an endangered neotropical primate facing habitat fragmentation pressure, the black lion tamarin (Leontopithecus chrysopygus). We investigated monthly and day-to-day glucocorticoid variations independently to attempt to disentangle the complex nature of the adrenocortical activity. Between May 2019 to March 2020, we followed two groups of black lion tamarins in two different areas, a continuous forest and a small fragment, and gathered behavioral data (over 95 days in total; 8.6 ± 3.9 days/month) and fecal samples (Nsamples = 468; 4.93 ± 3.5 samples/day) simultaneously. Preliminary analyses enabled us to identify circadian variations linked to the biological rhythm, which were taken into account in subsequent models. Monthly analyses revealed that black lion tamarin fecal glucocorticoid metabolite levels vary according to changes in activity budget associated with the fruit consumption, movement, and resting time of the groups. At a day-to-day level, while intergroup encounters led to increases in fecal glucocorticoid metabolite concentrations, we found that changes in food intake or activity level did not trigger physiological stress responses. These findings suggest that diet and ranging patterns, driven by food availability and distribution, influence physiological stress at a seasonal scale, while acute stressors such as interspecific competition trigger short-term stress responses. Exploring fecal glucocorticoid metabolite variations over different timescales can help uncover the predictive and reactive facets of physiological stress in wild species. Moreover, having a comprehensive understanding of the physiological state of species is a valuable conservation tool for evaluating how they cope in changing environments.

Unmanned aerial vehicles (UAVs) with thermal infrared (TIR) sensors are effective for monitoring and counting threatened Vietnamese primates.

Monitoring the population size of threatened primate species with minimal disturbance is becoming an outstanding requirement for conservation and wildlife management. Drones with thermal infrared (TIR) and visible spectrum (RGB) imaging are increasingly used to survey arboreal primates, but ground-truthing is still required to assess the effectiveness of drone-based count estimates. Our pilot study aims to assess the ability of a drone with both TIR and RGB sensors to detect, count, and identify semi-wild population of four endangered species of langurs and gibbon in the Endangered Primate Rescue Center (EPRC) in northern Vietnam. We found that TIR imagery enabled higher detection rates compared to RGB imagery and obtained an accurate count with the TIR only after four drone flights. We could identify langurs species based on thermal signature at a flight height of 50 m from the ground level (max tree height = 15 m), via size and shape of the body. With TIR imagery, we were able to record inconspicuous behaviors such as foraging and play. While some individuals initially showed flight or avoidance behaviors when the drone was sighted, these behaviors decreased or were absent on following drone surveys. Our study suggests that monitoring and precisely counting langur and gibbon species populations could be successful with the use of thermal drones only.

First survey on seroprevalence of Japanese encephalitis in long-tailed macaques (Macaca fascicularis) in Bali, Indonesia.

Background and Aim: Japanese encephalitis (JE) is a zoonotic infectious inflammatory brain disease caused by the JE virus (JEV). Considerable research into the seroprevalence of JE in domestic animals has been conducted, but there have been no reports of its occurrence in wild animals, including long-tailed macaques (Macaca fascicularis). This study aimed to estimate the seroprevalence of JEV infection and its determinants in long-tailed macaques in Bali and the prevalence of mosquito vectors. Materials and Methods: Blood samples (3 mL) were collected from a population of M. fascicularis (92 heads) inhabiting a small forest with irrigated rice field nearby (wetland area) in Ubud, Gianyar, and from two populations in dryland areas with no wet rice field (Uluwatu, Badung, and Nusa Penida, Bali Province, Indonesia). The collected sera were tested for antibodies against JEV using a commercially available enzyme-linked immunosorbent assay kit (qualitative monkey JE Immunoglobulin G antibody kit). The seropositivity of the antibodies was then compared based on different variables, namely, habitat type, age, and sex. Results: The seroprevalence of the JEV antibodies in all the samples tested was found to be 41.3%. The seropositivity of the monkey serum samples collected from the wetland area was 46.4%, which was higher than the seropositivity of the sera samples collected from the dried field areas (1.25%). Monkey sera collected from the wetland areas were 6.1 times (odds ratio [OR]: 6.1; 95% confidence interval [CI]: 0.71-51.5, p>0.05) more likely to be seropositive compared to the monkey sera collected from the dried field areas. Meanwhile, female monkeys were 1.79 times (OR: 1.79; 95% CI: 0.76-4.21; p>0.05) more likely to be seropositive to JEV than males. Similarly, juvenile monkeys were 2.38 times (OR: 2.38; 95% CI: 0.98-5.79); p>0.05) more likely to be seropositive against the JEV than adult monkeys. However, none of these differences achieved statistical significance. Regarding the JEV mosquito vector collection, more Culex mosquitoes were found in the samples from the wetland areas than from the dried field areas. Conclusion: The study confirms the existence of JEV infection in long-tailed macaques in Bali. There were patterned seropositivity differences based on habitat, age, and sex of the monkeys, but these were not significant. The possibility of monkeys as a JEV reservoir and the presence of the mosquitoes as the JEV vector are suggested but require more study to confirm.

First Descriptive Analysis of the Faecal Microbiota of Wild and Anthropized Barbary Macaques (Macaca sylvanus) in the Region of Bejaia, Northeast Algeria.

Previous research has revealed the gut microbiota profile of several primate species, as well as the impact of a variety of anthropogenic factors, such as tourist food supply, on these bacterial communities. However, there is no information on the gut microbiota of the endangered wild Barbary macaque (Macaca sylvanus). The present study is the first to characterize the faecal microbiota of this species, as well as to investigate the impact of tourist food provisioning on it. A total of 12 faecal samples were collected in two groups of M. sylvanus in the region of Bejaia in Algeria. The first group-a tourist-provisioned one-was located in the tourist area of the Gouraya National Park and the second group-a wild-feeding one-was located in the proximity of the village of Mezouara in the forest of Akfadou. After DNA extraction, the faecal microbiota composition was analysed using 16S rDNA sequencing. Statistical tests were performed to compare alpha diversity and beta diversity between the two groups. Non-metric multidimensional scaling analysis (NMDS) was applied to visualize biodiversity between groups. Behaviour monitoring was also conducted to assess the time allocated to the consumption of anthropogenic food by the tourist-provisioned group. Our results revealed the presence of 209 bacterial genera from 17 phyla in the faecal microbiota of Barbary macaques. Firmicutes was the most abundant bacterial phylum, followed by Bacteroidetes and Verrucomicrobia. On the other hand, the comparison between the faecal microbiota of the two study groups showed that tourism activity was associated with a significant change on the faecal microbiota of M.sylvanus, probably due to diet alteration (with 60% of feeding time allocated to the consumption of anthropogenic food). The potentially low-fibre diet at the tourist site adversely influenced the proliferation of bacterial genera found in abundance in the wild group such as Ruminococcaceae. Such an alteration of the faecal microbiota can have negative impacts on the health status of these animals by increasing the risk of obesity and illness and calls for special management measures to reduce the provisioning rate in tourist areas.

Effect of Infant Presence on Social Networks of Sterilized and Intact Wild Female Balinese Macaques (Macaca fascicularis).

Contraception is increasingly used to control wild animal populations. However, as reproductive condition influences social interactions in primates, the absence of new offspring could influence the females' social integration. We studied two groups of wild macaques (Macaca fascicularis) including females recently sterilized in the Ubud Monkey Forest, Indonesia. We used social network analysis to examine female grooming and proximity networks and investigated the role of infant presence on social centrality and group connectivity, while controlling for the fertility status (sterilized N = 14, intact N = 34). We compared the ego networks of females experiencing different nursing conditions (young infant (YI) vs. old infant (OI) vs. non-nursing (NN) females). YI females were less central in the grooming network than other females while being more central in proximity networks, suggesting they could keep proximity within the group to protect their infant from hazards, while decreasing direct grooming interactions, involving potential risks such as kidnapping. The centrality of sterilized and intact females was similar, except for the proximity network where sterilized females had more partners and a better group connectivity. These results confirm the influence of nursing condition in female macaque social networks and did not show any negative short-term effects of sterilization on social integration.

Tubectomy of Pregnant and Non-pregnant Female Balinese Macaques (Macaca Fascicularis) With Post-operative Monitoring.

Worldwide, primates, and humans increasingly share habitats and often enter in conflict when primates thrive in human-dominated environments, calling for special management measures. Reproductive control is increasingly used to manage population growth but very few monitoring data are available. Therefore, the efficiency and implications of such programs require a careful examination. In the context of a contraception program in wild female long-tailed macaques in Ubud, Bali, conducted over four successive campaigns between 2017 and 2019, including 140 females (i.e., 41.9% of the reproductive females of the population in 2019), modifications of an endoscopic tubectomy procedure, a permanent sterilization method, clinical evaluation of this method, and the post-operative monitoring results of the neutered females after release are described. This surgical approach was applicable for pregnant females: 28.6% of the treated females were pregnant at the time of the surgery. The procedure used a single lateral port to reach and cauterize both oviducts in non-pregnant as well as in early to mid-term pregnant females. Pregnant females nearer to term required a second lateral port to access both oviducts masked by the size of the gravid uterus. Moreover, bipolar thermocauterization was utilized successfully without resection to realize the tubectomy. The average duration of the laparoscopic surgery was 14 min for non-pregnant females and 22 min for pregnant females. Animals were released 3 h 22 min in average following their capture. This short holding time, recommended for free-ranging primates, was made possible by the minimal invasiveness of the sterilization approach. A laparoscopic post-operative evaluation conducted on two patients during the following campaign confirmed that the oviducts were definitely disrupted and no longer patent. Moreover, no new pregnancies in sterilized females were recorded during the 3-year observation period. The survival rate of the treated females 6 months after sterilization was high (96.3%) with no major post-operative complications clinically recorded. Among females that were pregnant during surgery, 81.1% were confirmed to experience term delivery. This study demonstrates the safety and efficiency of endoscopic tubectomy, even for pregnant females, as a mean of wild macaques' population control.

A meta-analysis of anthropogenic impacts on physiological stress in wild primates.

As humanity continues to alter the environment extensively, comprehending the effect of anthropogenic disturbances on the health, survival, and fitness of wildlife is a crucial question for conservation science. Many primate populations occupy suboptimal habitats prone to diverse anthropogenic disturbances that may be sources of acute and chronic stress. Quantification of glucocorticoid (GC) concentrations has repeatedly been used to explore the impact of disturbances on physiological stress. Although it is still debated, prolonged elevation of GC levels may impair reproduction, growth, and immune system activity of individuals. We quantified the effect of anthropogenic disturbances on physiological stress in primates with a global meta-analysis based on data from 26 articles, covering 24 distinct species in 13 different countries. Anthropogenic disturbances were classified into 6 distinct categories: habitat loss, habitat degradation, ongoing logging, hunting, tourism, and other human activities. We calculated effect sizes (Hedges' g) with the standardized mean difference in GC concentrations between primates affected by human activity and their undisturbed conspecifics. We ran random-effects models and subgroup analyses to estimate the overall effect as well as a cumulative effect size for each disturbance category. Overall, primates inhabiting sites subject to anthropogenic disturbances exhibited significantly higher GC levels (g = 0.60; 95% CI: 0.28-0.93). Habitat loss and hunting were overall associated with increased GC concentrations, whereas the cumulative effects of the other disturbances were not statistically significant. Biologically, high GC levels may increase fitness by enabling individuals to overcome the challenges linked to anthropogenic disturbances. However, primates in disturbed environments may have sustained elevated GC levels. To strengthen future research, it is necessary to control confounding factors systematically (e.g., diet, reproductive status, predatory pressure, and resource availability) and improve understanding of the link between GC levels and the health, fitness, and survival of animals.

Metaanálisis de los Impactos Antropogénicos sobre el Estrés Fisiológico de los Primates Salvajes Resumen Conforme la humanidad continúa alterando extensivamente el ambiente, comprender el efecto de las perturbaciones antropogénicas sobre la salud, supervivencia y la aptitud de la fauna silvestre es una cuestión muy importante para las ciencias de la conservación. Muchas poblaciones de primates ocupan hábitats subóptimos propensos a varias perturbaciones antropogénicas que pueden ser la fuente de estrés agudo y crónico. La cuantificación de las concentraciones de glucocorticoides se ha usado repetidamente para explorar el impacto de las perturbaciones sobre el estrés fisiológico. Aunque todavía es tema de debate, la elevación prolongada de los niveles de glucocorticoides puede afectar a la reproducción, el crecimiento y la actividad del sistema inmune de los individuos. Cuantificamos el efecto de las perturbaciones antropogénicas sobre el estrés fisiológico de los primates mediante un metaanálisis global basado en los datos tomados de 26 artículos, abarcando así a 24 especies distintas en 13 países diferentes. Las perturbaciones antropogénicas fueron clasificadas en seis categorías distintas: pérdida de hábitat, degradación del hábitat, explotación forestal en curso, cacería, turismo y otras actividades humanas. Calculamos los tamaños del efecto (la g de Hedges) con una diferencia media estandarizada en las concentraciones de glucocorticoides entre los primates afectados por la actividad humana y sus coespecíficos sin perturbaciones. Corrimos modelos de efectos al azar y análisis de subgrupos para estimar el efecto generalizado, así como un tamaño acumulativo de efecto para cada categoría de perturbación. En general, los primates que habitan sitios sujetos a alguna perturbación antropogénica exhibieron niveles de glucocorticoides significativamente más altos (g = 0.60, 95% CI 0.28 a 0.93). La pérdida de hábitat y la cacería fueron asociadas casi siempre con las concentraciones elevadas de glucocorticoides, mientras que los efectos acumulativos de las otras perturbaciones no fueron significativos estadísticamente. Biológicamente, los niveles altos de glucocorticoides pueden incrementar la aptitud al permitir que el individuo supere los retos relaciones con las perturbaciones antropogénicas. Sin embargo, los primates en los ambientes perturbados pueden tener niveles elevados sostenidos de glucocorticoides. Para fortalecer a las futuras investigaciones es necesario controlar sistemáticamente los factores desconcertantes (p. ej.: la dieta, el estado reproductivo, la presión por depredadores, la disponibilidad de recursos) y mejorar el entendimiento de la conexión entre los niveles de glucocorticoides y la salud, aptitud y supervivencia de los animales.

Northern pigtailed macaques rely on old growth plantations to offset low fruit availability in a degraded forest fragment.

Space-use and foraging strategies are important facets to consider in regard to the ecology and conservation of primates. For this study, we documented movement, ranging, and foraging patterns of northern pigtailed macaques (Macaca leonina) for 14 months in a degraded habitat with old growth Acacia and Eucalyptus plantations at the Sakaerat Biosphere Reserve in northeastern Thailand. We used hidden Markov models and characteristic hull polygons to analyze these patterns in regard to fruit availability. Macaques' home range (HR) was 599 ha and spanned through a natural dry-evergreen forest (DEF), and plantation forest. Our results showed that active foraging increased with higher fruit availability in DEF. Macaques changed to a less continuous behavioral state during periods of lower fruit availability in DEF, repeatedly moving from foraging to transiting behavior, while extending their HR further into plantation forest and surrounding edge areas. Concomitantly, macaques shifted their diet from fleshy to dry fruit such as the introduced Acacia species. Our results showed that the diet and movement ecology adaptations of northern pigtailed macaques were largely dependent on availability of native fruits, and reflected a "high-cost, high-yield" foraging strategy when fresh food was scarce and dry fruit was available in plantation forest. Conversely, wild-feeding northern pigtailed macaque populations inhabiting pristine habitat approached a "low-cost, low-yield" foraging strategy. Our results outline the effects of habitat degradation on foraging strategies and show how a flexible species can cope with its nutritional requirements.

Social influence on the expression of robbing and bartering behaviours in Balinese long-tailed macaques.

Animals use social information, available from conspecifics, to learn and express novel and adaptive behaviours. Amongst social learning mechanisms, response facilitation occurs when observing a demonstrator performing a behaviour temporarily increases the probability that the observer will perform the same behaviour shortly after. We studied "robbing and bartering" (RB), two behaviours routinely displayed by free-ranging long-tailed macaques (Macaca fascicularis) at Uluwatu Temple, Bali, Indonesia. When robbing, a monkey steals an inedible object from a visitor and may use this object as a token by exchanging it for food with the temple staff (bartering). We tested whether the expression of RB-related behaviours could be explained by response facilitation and was influenced by model-based biases (i.e. dominance rank, age, experience and success of the demonstrator). We compared video-recorded focal samples of 44 witness individuals (WF) immediately after they observed an RB-related event performed by group members, and matched-control focal samples (MCF) of the same focal subjects, located at similar distance from former demonstrators (N = 43 subjects), but in the absence of any RB-related demonstrations. We found that the synchronized expression of robbing and bartering could be explained by response facilitation. Both behaviours occurred significantly more often during WF than during MCF. Following a contagion-like effect, the rate of robbing behaviour displayed by the witness increased with the cumulative rate of robbing behaviour performed by demonstrators, but this effect was not found for the bartering behaviour. The expression of RB was not influenced by model-based biases. Our results support the cultural nature of the RB practice in the Uluwatu macaques.

Foraging strategies underlying bird egg predation by macaques: A study using artificial nests.

Bird egg predation is widespread in non-human primates. Although nest predation is often described as opportunistic, little is known about foraging strategies and nest detection in primates. Since it is the prevalent cause of nest failure in the tropics, birds select nest sites within specific microhabitats and use different nest types to increase nesting success. Identifying the nests targeted by the northern pigtailed macaques (Macaca leonina), an omnivorous cercopithecine species, and known nest predator, will shine light on nest foraging strategies in primates. The aim of this research was to reveal if nest predation is a selective or opportunistic feeding behavior. We studied, using artificial nests and camera traps, the influence of nest type (open-cup vs. cavity), microhabitat (i.e., understory density, canopy cover, canopy height, ground cover, and presence vs. absence of thorns and lianas), and nest height, on nest predation by a troop of northern pigtailed macaques in the Sakaerat Biosphere Reserve (Thailand), a degraded environment. In our study, macaque predation on artificial nests was high; out of the 200 nests that were set up, 112 were plundered by macaques. Although predation rates decreased with nest height, nest type, and microhabitat had no significant effect on predation by macaques. Nest detectability and accessibility did not affect predation rates. Macaques actively searched for nests in different microhabitats, suggesting that nest predation by this primate might be considered a selective feeding behavior in this degraded habitat. Consequently, nest predation by this primate might have important conservation implications on the population dynamics of forest-dwelling bird species. Behavior observation methods, such as instantaneous scan sampling, may underestimate nest predation by primates, a furtive and cryptic behavior.

Intergroup variation in robbing and bartering by long-tailed macaques at Uluwatu Temple (Bali, Indonesia).

Robbing and bartering (RB) is a behavioral practice anecdotally reported in free-ranging commensal macaques. It usually occurs in two steps: after taking inedible objects (e.g., glasses) from humans, the macaques appear to use them as tokens, returning them to humans in exchange for food. While extensively studied in captivity, our research is the first to investigate the object/food exchange between humans and primates in a natural setting. During a 4-month study in 2010, we used both focal and event sampling to record 201 RB events in a population of long-tailed macaques (Macaca fascicularis), including four neighboring groups ranging freely around Uluwatu Temple, Bali (Indonesia). In each group, we documented the RB frequency, prevalence and outcome, and tested the underpinning anthropogenic and demographic determinants. In line with the environmental opportunity hypothesis, we found a positive qualitative relation at the group level between time spent in tourist zones and RB frequency or prevalence. For two of the four groups, RB events were significantly more frequent when humans were more present in the environment. We also found qualitative partial support for the male-biased sex ratio hypothesis [i.e., RB was more frequent and prevalent in groups with higher ratios of (sub)adult males], whereas the group density hypothesis was not supported. This preliminary study showed that RB is a spontaneous, customary (in some groups), and enduring population-specific practice characterized by intergroup variation in Balinese macaques. As such, RB is a candidate for a new behavioral tradition in this species.

The role of anthropic, ecological, and social factors in sleeping site choice by long-tailed macaques (Macaca fascicularis).

When choosing their sleeping sites, primates make adaptive trade-offs between various biotic and abiotic constraints. In human-modified environments, anthropic factors may play a role. We assessed the influence of ecological (predation), social (intergroup competition), and anthropic (proximity to human settlements) factors in sleeping site choice by long-tailed macaques (Macaca fascicularis) occupying a habitat at the interface of natural forests and human-modified zones in Bali Barat National Park, Indonesia. Over the course of 56 nights, we collected data relating to physical features of sleeping trees, patterns of the use of sleeping sites within the home range, pre-sleep behavior, diurnal ranging patterns and availability of natural and human food. Overall, the macaques used 17 sleeping sites with 37 sleeping trees. When the monkeys slept in forest zones, they selected sleeping trees that had larger trunks but were not significantly taller than surrounding trees. Though the macaques rarely re-used sleeping sites on consecutive nights, they frequently re-used four sites over the study period. The group favored sleeping within the core area of its home range, despite the occurrence of frequent agonistic intergroup encounters there. Macaques preferentially selected sleeping trees located within or near human-modified zones, especially when human food was abundant and natural food was scarce. These results partially support the hypothesis that long-tailed macaques choose their sleeping sites to avoid predation; proximity to human settlements appears to be the primary factor influencing sleeping site choice in this primate species. Our results reflect the strong influence that anthropic factors have on primates, which subsist in increasingly human-dominated landscapes.