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

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

Pygmy Marmoset Exudate Feeding Stimulates Exudate Production.

Exudates are an important renewable resource for many primates. Exudate renewability is based on observations of primates repeatedly depleting exudate sites and measures of exudate trees' daily replenishment rates, but the role of the consumer in the renewal process is unclear. Trees' exudate production may be independent of the consumer, remaining unchanged regardless of depletion frequency, but since trees produce exudates as a physiological response to fungal infection, they may produce more exudates with more frequent depletion. To test these competing hypotheses, we employed a within-subjects experimental design in which we exposed pygmy marmosets' exudate holes to two treatment conditions: collecting exudates after 5 h and collecting exudates every hour for five consecutive hours. To compare production outputs between treatments, we used generalised linear mixed modelling in which log-transformed production data were a function of treatment with exudate holes nested within trees as a random effect. The model indicated that the cumulative production of hourly exudate removal was significantly greater than the amount accumulated after 5 h. Furthermore, the random effect of holes nested within trees had the greatest impact on variation in differences between treatments, but another unknown source also contributed to the observed variation. These results support the hypothesis that consumers partly drive exudate production, and although it is unknown what other factors, such as fungal load and healing trajectory, may influence variation between treatments, we conclude that pygmy marmosets can stimulate exudate production by consuming exudates.

Reproductive seasonality in wild northern pig-tailed macaques (Macaca leonina).

Macaque reproductive patterns range from strictly seasonal breeding to non-seasonal breeding, but factors explaining this variation are not fully understood. Valid reproductive seasonality data are also still lacking for many wild macaque populations because the majority of birth data are from captive animals living outside of their geographic range. We evaluated whether the reproductive seasonality of wild northern pig-tailed macaques falls as expected by the ecological (latitude) or phylogenetic inertia hypotheses in comparison with other macaque species. We recorded monthly occurrences of births (N = 22), copulations (N = 563), and females exhibiting sex skin swellings (N = 18) in one group at Khao Yai National Park (KYNP), Thailand. Births, copulations, and females exhibiting sex skin swellings were significantly different from a random distribution. Using measures of circular statistics and the van Schaik and colleagues' (Schaik et al. Lee (ed), Comparative primate socioecology, Cambridge University Press, Cambridge, 1999) seasonality categorizations, the population at KYNP is best characterized as moderately seasonal breeding. Despite some inconsistency, macaque reproductive seasonality was significantly influenced by latitudinal location. We broadly found that: (1) non-seasonal breeding macaque populations (birth r-vector < 0.3) lived at latitudes close to the equator between 1°S and 3°N, (2) moderately seasonal breeding macaque populations (0.3 < birth r-vector < 0.7) were found between 3°N and 14°N and at 5°S, and (3) strictly seasonal breeding macaque populations (birth r-vector > 0.7) ranged ≥ 12°N. A strong phylogenetic signal in reproductive seasonality on the macaque phylogeny was also detected. However, further studies of wild macaque populations are still needed to better characterize reproductive seasonality in this taxon.

White-handed gibbons discriminate context-specific song compositions.

White-handed gibbons produce loud and acoustically complex songs when interacting with their neighbours or when encountering predators. In both contexts, songs are assembled from a small number of units although their composition differs in context-specific ways. Here, we investigated whether wild gibbons could infer the 'meaning' when hearing exemplars recorded in both contexts (i.e. 'duet songs' vs. 'predator songs'). We carried out a playback experiment by which we simulated the presence of a neighbouring group producing either its duet or a predator song in order to compare subjects' vocal and locomotor responses. When hearing a recording of a duet song, subjects reliably responded with their own duet song, which sometimes elicited further duet songs in adjacent groups. When hearing a recording of a predator song, however, subjects typically remained silent, apart from one of six groups which replied with its own predator song. Moreover, in two of six trials, playbacks of predator songs elicited predator song replies in non-adjacent groups. Finally, all groups showed strong anti-predator behaviour to predator songs but never to duet songs. We concluded that white-handed gibbons discriminated between the two song types and were able to infer meaning from them. We discuss the implications of these findings in light of the current debate on the evolutionary origins of syntax.

Facial expressions and pair bonds in hylobatids.

OBJECTIVES: Facial expressions are an important component of primate communication that functions to transmit social information and modulate intentions and motivations. Chimpanzees and macaques, for example, produce a variety of facial expressions when communicating with conspecifics. Hylobatids also produce various facial expressions; however, the origin and function of these facial expressions are still largely unclear. It has been suggested that larger facial expression repertoires may have evolved in the context of social complexity, but this link has yet to be tested at a broader empirical basis. The social complexity hypothesis offers a possible explanation for the evolution of complex communicative signals such as facial expressions, because as the complexity of an individual's social environment increases so does the need for communicative signals. We used an intraspecies, pair-focused study design to test the link between facial expressions and sociality within hylobatids, specifically the strength of pair-bonds. MATERIAL AND METHODS: The current study compared 206 hr of video and 103 hr of focal animal data for ten hylobatid pairs from three genera (Nomascus, Hoolock, and Hylobates) living at the Gibbon Conservation Center. Using video footage, we explored 5,969 facial expressions along three dimensions: repertoire use, repertoire breadth, and facial expression synchrony [FES]. We then used focal animal data to compare dimensions of facial expressiveness to pair bond strength and behavioral synchrony. RESULTS: Hylobatids in our study overlapped in only half of their facial expressions (50%) with the only other detailed, quantitative study of hylobatid facial expressions, while 27 facial expressions were uniquely observed in our study animals. Taken together, hylobatids have a large facial expression repertoire of at least 80 unique facial expressions. Contrary to our prediction, facial repertoire composition was not significantly correlated with pair bond strength, rates of territorial synchrony, or rates of behavioral synchrony. We found that FES was the strongest measure of hylobatid expressiveness and was significantly positively correlated with higher sociality index scores; however, FES showed no significant correlation with behavioral synchrony. No noticeable differences between pairs were found regarding rates of behavioral or territorial synchrony. Facial repertoire sizes and FES were not significantly correlated with rates of behavioral synchrony or territorial synchrony. DISCUSSION: Our study confirms an important role of facial expressions in maintaining pair bonds and coordinating activities in hylobatids. Data support the hypothesis that facial expressions and sociality have been linked in hylobatid and primate evolution. It is possible that larger facial repertoires may have contributed to strengthening pair bonds in primates, because richer facial repertoires provide more opportunities for FES which can effectively increase the "understanding" between partners through smoother coordination of interaction patterns. This study supports the social complexity hypothesis as the driving force for the evolution of complex communication signaling.

Short dispersal distance of males in a wild white-handed gibbon (Hylobates lar) population.

OBJECTIVES: It has long been recognized that in gibbons both sexes disperse from the natal group. However, the fate of dispersed individuals was rarely documented. Here we provide the first detailed information on sex differences in dispersal patterns by analyzing the spatial genetic structure of a well-known white-handed gibbon (Hylobates lar) population. MATERIALS AND METHODS: Mitochondrial DNA (mtDNA) and Y-chromosomal haplotypes, and autosomal microsatellite genotypes were determined for individuals of the Mo Singto study site, Khao Yai National Park, Thailand. Mantel tests for the three genetic marker types were performed for 17 gibbon groups comprising 23 adult males and 18 adult females. RESULTS: Significant positive Mantel correlations were observed for spatial distance and both autosomal microsatellite-based as well as Y-chromosomal haplotype-based genetic distance among adult males. Neighboring adult males tended to be genetically related and share Y-chromosomal haplotypes. Conversely, no significant Mantel correlations were observed either in autosomal microsatellites or mtDNA among adult females. DISCUSSION: Our results confirm, at a genetic level, hypotheses from long-term demographic observations that white-handed gibbon males of the Mo Singto population primarily disperse into adjacent groups. Instead, females disperse more opportunistically either to adjacent or more distant groups. This sex-specific difference reflects an apparent greater tolerance between males than between females. The higher tolerance of adult males allows the formation of stable multimale groups and facilitates male dispersal into an adjacent group. Stable multifemale groups have never been documented for white-handed gibbons probably due to feeding competition between females.

Extractive foraging of toxic caterpillars in wild northern pig-tailed macaques (Macaca leonina).

Extractive foraging in nonhuman primates may involve different levels of technical complexity in terms of the number of actions that must be performed and the manual dexterity involved. We describe the extractive foraging of caterpillars in wild northern pig-tailed macaques (Macaca leonina) at Khao Yai National Park, Thailand. The study group, observed from May to December 2016 (n = 146 days), comprised 60-70 habituated individuals, including 3-4 adult males, 20-23 adult females, and 36-47 immatures. Four adult males and five adult females, observed from September to November 2016 for a total of 24 days, were selected for focal animal sampling. Northern pig-tailed macaques were observed eating at least two families (Erebidae and Limacodidae) and three genera (Macrobrochis sp., Phlossa sp. and Scopelodes sp.) of caterpillars. While the monkeys ate short and small caterpillars with stinging setae and non-setae caterpillars without processing, they performed extensive caterpillar-rubbing behavior on large and long caterpillars with stinging setae. Based on 61 extractive foraging bouts, we found that caterpillar rubbing was hierarchically organized into five stages and 12 elements. Five stages of behavior sequence started with picking the caterpillar up, transporting it to a substrate, rubbing it to remove stinging setae, ingesting it, and then cleaning hands and mouth. Only adult macaques were observed using a leaf to rub stinging caterpillars.

Context-specific close-range "hoo" calls in wild gibbons (Hylobates lar).

BACKGROUND: Close range calls are produced by many animals during intra-specific interactions, such as during home range defence, playing, begging for food, and directing others. In this study, we investigated the most common close range vocalisation of lar gibbons (Hylobates lar), the 'hoo' call. Gibbons and siamangs (family Hylobatidae) are known for their conspicuous and elaborate songs, while quieter, close range vocalisations have received almost no empirical attention, perhaps due to the difficult observation conditions in their natural forest habitats. RESULTS: We found that 'hoo' calls were emitted by both sexes in a variety of contexts, including feeding, separation from group members, encountering predators, interacting with neighbours, or as part of duet songs by the mated pair. Acoustic analyses revealed that 'hoo' calls varied in a number of spectral parameters as a function of the different contexts. Males' and females' 'hoo' calls showed similar variation in these context-specific parameter differences, although there were also consistent sex differences in frequency across contexts. CONCLUSIONS: Our study provides evidence that lar gibbons are able to generate significant, context-dependent acoustic variation within their main social call, which potentially allows recipients to make inferences about the external events experienced by the caller. Communicating about different events by producing subtle acoustic variation within some call types appears to be a general feature of primate communication, which can increase the expressive power of vocal signals within the constraints of limited vocal tract flexibility that is typical for all non-human primates. In this sense, this study is of direct relevance for the on-going debate about the nature and origins of vocally-based referential communication and the evolution of human speech.

Extra-pair paternity confirmed in wild white-handed gibbons.

Knowledge of the genetic mating system of animal species is essential for our understanding of the evolution of social systems and individual reproductive strategies. In recent years, genetic methods have uncovered an unexpected diversity of paternal genetic contributions across diverse animal social mating systems, but particularly in pair-living species. In most pair-living birds, for example, genetic and behavioral observations have confirmed a previously unknown significance of extra-pair copulations (EPCs) and extra-pair paternity. Among mammals, white-handed gibbons (Hylobates lar) are also known to live in pairs and are traditionally believed to be single-male single-female breeders. However, at Khao Yai National Park, Thailand, behavioral observations have confirmed the occurrence of both EPCs and functional multi-male grouping, but knowledge about the genetic mating system is still unavailable. In this study, we genotyped 89 white-handed gibbons of the Khao Yai population based on fecal samplings and were able to determine paternity for 41 offspring through short tandem repeat analysis. We found that females' stable social partners sired the majority (90.5%) of offspring (N = 38), while only a few (7.1%) offspring (n = 2 confirmed cases; n = 1 inferred case) were conceived with extra-pair partners. The paternity of one offspring remained inconclusive (2.4%), because the offspring's genotype did not mismatch with the genotypes of two potential sires. Like other predominantly pair-living species, gibbons appear to follow a mixed-reproductive strategy. The genetic mating system of wild white-handed gibbons is best described as flexible, primarily monogamous and opportunistically promiscuous. Inc.

Molecular evidence for the introgression between Hylobates lar and H. pileatus in the wild.

Inter-specific hybrid zones for Hylobates gibbons are known in Southeast Asia. Among these, one hybrid zone between Hylobates lar and H. pileatus is located in Khao Yai National Park, Thailand. To find molecular evidence for the natural hybridization of the gibbons in this region, we studied mitochondrial DNA (mtDNA) of 68 gibbons of the H. lar phenotype living adjacent to the hybrid zone. Nucleotide sequencing of a fragment of mtDNA spanning hyper variable segment I showed that nine gibbons had an mtDNA haplotype of H. pileatus, and that seven of these nine gibbons belonged to a single maternal lineage over three generations. It is thus confirmed that introgression between H. lar and H. pileatus exists and the initial hybridization took place ages ago.

Gibbon travel paths are goal oriented.

Remembering locations of food resources is critical for animal survival. Gibbons are territorial primates which regularly travel through small and stable home ranges in search of preferred, limited and patchily distributed resources (primarily ripe fruit). They are predicted to profit from an ability to memorize the spatial characteristics of their home range and may increase their foraging efficiency by using a 'cognitive map' either with Euclidean or with topological properties. We collected ranging and feeding data from 11 gibbon groups (Hylobates lar) to test their navigation skills and to better understand gibbons' 'spatial intelligence'. We calculated the locations at which significant travel direction changes occurred using the change-point direction test and found that these locations primarily coincided with preferred fruit sources. Within the limits of biologically realistic visibility distances observed, gibbon travel paths were more efficient in detecting known preferred food sources than a heuristic travel model based on straight travel paths in random directions. Because consecutive travel change-points were far from the gibbons' sight, planned movement between preferred food sources was the most parsimonious explanation for the observed travel patterns. Gibbon travel appears to connect preferred food sources as expected under the assumption of a good mental representation of the most relevant sources in a large-scale space.

Sterile pyuria in a population of wild white-handed gibbons (Hylobates lar).

Urinalysis is an emerging method for monitoring the health and energy balance of wild primates. Here, we report the first urinalysis of wild gibbons. We used multi-reagent test strips to monitor the health status of 52 individual white-handed gibbons (Hylobates lar) inhabiting Khao Yai National Park, Thailand. Most urinary reference values were within normal ranges; however, regardless of age- and sex-class or monthly fruit productivity, we found unexpectedly high rates of urinary leukocytes (50% and 90% of individuals in 2001-2003 and 2006, respectively). In contrast to previous studies of African apes, this finding is coupled with the near absence of urinary nitrites, demonstrating pervasive levels of sterile pyuria. This result is the first reported case of sterile pyuria in a population of wild primates. The etiology of human sterile pyuria is diverse, but in all cases it is diagnostic of systemic inflammation. We discuss the potential causes of sterile pyuria in the gibbons of Khao Yai.

Home-range characteristics and the influence of seasonality on female reproduction in white-handed gibbons (Hylobates lar) at Khao Yai National Park, Thailand.

A three-year (2001-2003) study was carried out on the home range characteristics of seven wild white-handed gibbon (Hylobates lar) groups focusing on the spatio-temporal distribution of food resources at Khao Yai National Park in northeastern Thailand. These results were combined with 23 years (1980-2003) of reproductive performance data on seven females from the same focal groups. Reproductive performance was equal among females with regard to birth, weaning and maturation ratios, and independent of variation in food availability. Offspring mortality, however, was significantly positively correlated with home-range size. In addition, there was an increase in offspring mortality just after weaning, suggesting that the increase in the daily distance traveled by juveniles contributed to this mortality. Conceptions clustered during the first half of the year when food production was at its peak, which presumably allowed females to accumulate sufficient body reserves to resume ovarian cycling. Our results place Khao Yai gibbons closer to Cercopithecidae than great apes in terms of the temporal pattern of reproductive events, though gestation, lactation, inter-birth interval, and offspring maturation are considerably longer in gibbons, placing them closer to the other apes. Our findings underline the unique phylogenetic position of these small-bodied apes in terms of reproductive patterns in primates.

Sexual swellings in wild white-handed gibbon females (Hylobates lar) indicate the probability of ovulation.

Conspicuous sexual swellings in the females of some primate species have been a focus of scientific interest since Darwin first wrote about them in 1871. To understand these visual signals, research focused on exaggerated sexual swellings of Old World primates. However, some primate species develop much smaller sexual swellings and it is as yet unclear if these smaller swellings can serve similar functions as those proposed for exaggerated swellings, i.e. advertising fertility to attract mates. We studied the temporal patterns of sexual swellings, timing of ovulation and female reproductive status in wild white-handed gibbons (Hylobates lar) at Khao Yai National Park, Thailand, where this species has a variable social organization. We established fecal progestogen profiles in fifteen cycles of eight cycling females and, to detect swellings outside the menstrual cycle, five pregnant and six lactating females. In 80% of menstrual cycles, ovulation and maximum swelling phase (duration: Ø 9.3 days; 42.8% of cycle length), overlapped tightly. The probability of ovulation peaked on day 3 of the maximum swelling period. Nevertheless, the temporal relationship between maximum swelling and probability of ovulation varied from day -1 to day 13 of the swelling period and three times ovulations fell outside the maximum swelling phase. The different swellings phases occurred in similar proportions in cycling and pregnant, but not lactating females, which were rarely swollen. Despite their smaller size, gibbons' sexual swellings probably serve functions similar to those suggested for exaggerated swellings by the graded-signal hypothesis, which predicts that sexual swellings indicate the probability of ovulation, without allowing males to pinpoint its exact time.

The syntax and meaning of wild gibbon songs.

Spoken language is a result of the human capacity to assemble simple vocal units into more complex utterances, the basic carriers of semantic information. Not much is known about the evolutionary origins of this behaviour. The vocal abilities of non-human primates are relatively unimpressive in comparison, with gibbon songs being a rare exception. These apes assemble a repertoire of call notes into elaborate songs, which function to repel conspecific intruders, advertise pair bonds, and attract mates. We conducted a series of field experiments with white-handed gibbons at Khao Yai National Park, Thailand, which showed that this ape species uses songs also to protect themselves against predation. We compared the acoustic structure of predatory-induced songs with regular songs that were given as part of their daily routine. Predator-induced songs were identical to normal songs in the call note repertoire, but we found consistent differences in how the notes were assembled into songs. The responses of out-of-sight receivers demonstrated that these syntactic differences were meaningful to conspecifics. Our study provides the first evidence of referential signalling in a free-ranging ape species, based on a communication system that utilises combinatorial rules.

Cross-species amplification of human microsatellite markers using noninvasive samples from white-handed gibbons (Hylobates lar).

Analysis of the population genetic structure and reproductive strategies of various primate species has been facilitated by cross-species amplification (i.e., the use of microsatellite markers developed in one species for analysis of another). In this study we screened 47 human-derived markers to assess their utility in the white-handed gibbon (Hylobates lar). Only eight produced accurate, reliable results, and exhibited levels of polymorphism that were adequate for individual identification. This low success rate was surprising given that human microsatellite markers typically work well in species (such as macaques) that are evolutionarily more distant from humans than are gibbons. In addition, we experienced limited success in using a set of microsatellite markers that have been reported to be useful in the closely-related H. muelleri, and applying our set of microsatellite markers to samples obtained from one H. pileatus individual. Our results emphasize the importance of extensively screening potential markers in representatives of the population of interest.