Phenotypic plasticity enables animals to adjust physiology, behaviour, morphology and life-history traits in response to changing conditions, either reversibly or through irreversible developmental shifts. In long-lived species, early-life phenotypic changes can have profound consequences if they persist into adulthood. Understanding the balance between stable and flexible trait expression across ontogeny is therefore key. Glucocorticoids (GCs) are central to physiological regulation and known to exhibit plasticity, but little is known about the consistency of GC phenotypes across development. We examined whether bonobos (Pan paniscus), a long-lived species, show consistent GC phenotypes as they mature, focusing on individual differences in average urinary cortisol phenotypes (reaction-norm intercepts) and plasticity (reaction-norm slopes) in response to time of day. We applied a reaction-norm approach to assess individual variation in GC intercepts and slopes across ontogeny, using random regression mixed-effects models. Trait repeatability of urinary cortisol was low across and within years, indicating high within-individual variation relative to between-individual variation. Reaction-norm intercepts were moderately repeatable, suggesting stable individual average GC phenotypes across development. By contrast, slopes were weakly repeatable, reflecting flexibility in how individuals modulate GC output across the day. This dual regulatory structure may support adaptive physiological responses to changing demands in a long-lived species.

Most mammals, including humans, exhibit even or slightly male-biased birth sex ratios (BSRs) and female-biased adult sex ratios (ASRs) much later in life due to higher male mortality rates. The group-living primates of Madagascar are unusual in this respect because they lack female-biased ASRs, but it is unknown whether this is the result of skewed BSRs or sex-specific disappearance patterns. Using long-term demographic data from wild red-fronted lemurs ( Eulemur rufifrons ), we analysed their sex ratio dynamics across the lifespan. We assessed BSR via prenatal sex determination using maternal faecal oestrogen metabolite measurements during late pregnancy, confirming a visually determined equal sex ratio three months after birth, and indicating no early sex-specific mortality. Demographic analyses additionally disclosed higher female disappearance within the first 8 years of age, likely associated with reproductive effort early in life. Thereby, adult male survival had the greatest positive effect on the ASR. Our study offers a rare perspective on the dynamics of age- and sex-specific disappearance in a wild primate population, whose sex-reversed patterns may also contribute to a more general understanding of the mechanisms generating sex-biased mortality.

The hypothalamic-pituitary-adrenal (HPA) axis plays a dual role in the biology of developmental plasticity in mammals, including humans—HPA axis activity not only provides the input for, but is also a target of, offspring developmental plasticity. To investigate the understudied effects of exposure timing, this study quantified maternal HPA axis activity during each half of gestation as well as during early lactation and assessed its effect on offspring HPA axis activity in a cross-sectional sample of infant, juvenile and adult Assamese macaques ( Macaca assamensis ). To add ecological validity to experimental studies under laboratory conditions, macaques were studied in the wild. Increased maternal faecal glucocorticoid (GC) metabolite levels experienced early in gestation, but not postnatal exposure during lactation were associated with increased offspring HPA axis activity from infancy into adulthood. Building on prior findings, this study indicates that significant timing effects not only influence the presence, magnitude and direction, but also the consistency of maternal GC effects on offspring HPA axis function.

In mammals, estrogens and progestogens are crucial for gestation, fetal development, and maternal preparation for parturition and lactation. Measuring these hormones allows for the diagnosis of pregnancy, estimation of pregnancy failures, and potentially prenatal sex determination. We evaluated urinary estrogen and progestogen metabolites as biomarkers for gestation detection and for their utility for fetal sex determination in wild Assamese macaques (Macaca assamensis) using 586 samples from 19 females, including 19 successful pregnancies. Four enzyme-immunoassays were tested for suitability in measuring urinary sex steroids using serial dilution: three assays targeting progestogen and one targeting estrogen metabolites (estrone conjugates, E1C). We performed a biological validation by measuring urinary hormone metabolites in one female across pre-, early-, late-, and post-gestation. None of the progestogen measurements reflected gestational status, while E1C levels showed the expected increases during gestation. Next, we measured urinary E1C across gestation in all females and investigated fetal sex effects on maternal E1C levels, expecting differences between females carrying male versus female fetuses. Urinary E1C levels increased as early as 9 days postconception and declined sharply at parturition, mirroring patterns in other primates. During late gestation, females carrying male fetuses had significantly higher E1C levels than those carrying female fetuses, yet overlapping values limit precision for prenatal sex determination. Urinary E1C offers a noninvasive marker for gestation monitoring in Assamese macaques, with application in ecological and conservation research. Additionally, results indicate intra- and inter-species-specific differences in steroid hormone metabolism and excretion, which need to be considered when selecting markers for reproductive monitoring.

Intraspecific competition with fellow group members represents an unavoidable cost of group living. However, the causes of competition can vary among group members, and ecological and reproductive challenges faced by individuals throughout the year can trigger physical conflicts and or physiological responses. To date, few studies in mammals have described both physiological and behavioral responses to competition simultaneously across the year in both males and females. However, such an approach may shed light on ultimate drivers of sex-specific competitive strategies. In this six-year study on multiple groups of wild redfronted lemurs (Eulemur rufifrons), a primate species from Madagascar, we intended to identify the relative importance of feeding vs. reproductive competition for both sexes. We combined data on fecal glucocorticoid metabolite (FGCM) levels, a proxy for the physiological stress response, with behavioral observations on agonistic interactions during ecologically and socially challenging phases across the year. We found that while FGCM levels increased in both sexes with decreasing fruit consumption, this increase was not accompanied by concomitant changes in agonistic behavior. Female aggression and FGCM levels instead peaked during the birth season, while for males, aggression remained fairly constant across the year. Our results suggest that redfronted lemurs have mechanisms to avoid direct competition through aggression at times when individuals may need to conserve energy.

Background The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial. Here, we compared the efficacy of two hormonal ovarian stimulation approaches: 1) stimulation of follicular growth with hFSH followed by triggering of oocyte maturation with hCG (FSH + hCG) and 2) stimulation with hFSH only (FSH-priming). Methods In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates. Results Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes. Conclusion A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.

Strong social bonds in gregarious adult animals have been associated with lower levels of glucocorticoids. However, similar research is lacking for juvenile primates. We examined relationships between social bonds and mean concentrations of fecal glucocorticoid metabolites (fGCMs) in 44 free-ranging juvenile rhesus macaques (Macaca mulatta) on Cayo Santiago, Puerto Rico. We measured frequencies of affiliative behavior (grooming, play, approaches and proximity) with other same-sex, same-aged juveniles (peers) and the total number of affiliative peer relationships. We found a positive relationship between fGCMs and grooming frequencies. Females that spent more time in proximity to peers also had higher fGCMs. In contrast, among juveniles with more closely related peers, those with more affiliative peers or more frequent play bouts had lower fGCMs. However, strong peer bonds in most juveniles did not appear to be associated with reduced glucocorticoid levels. fGCMs were higher for females than males, but were unassociated with physical activity, aggression, or peer seeking tendencies. We propose that the establishment and navigation of some peer bonds at this life stage may involve increased metabolic demand.

The measurement of biomarkers in blood and excreta can enable immune status assessment and provide prognostic information on individual health outcomes. In this respect, the fecal measurement of secretory immunoglobulin A (sIgA), the primary mammalian antibody for mucosal defense, has recently received increased interest in a few anthropoid primates, but a fecal sIgA assay for use in strepsirrhine primates has not yet been reported. Here, we develop and analytically validate a cost-effective in-house sandwich enzyme immunoassay for the extraction and measurement of sIgA in feces of redfronted lemurs (Eulemur rufifrons). We also tested a simple method for sIgA extraction that can be used under remote field conditions and undertook experiments to assess the robustness of sIgA concentrations to variation in processing and storage conditions of fecal extracts. Our analytical validation revealed that the assay recognizes immunoreactive sIgA in redfronted lemur feces, that sIgA can be measured accurately with no potential interference from the fecal matrix, and that assay reagents and performance are highly stable over time. The field-friendly extraction procedure produced sIgA results strongly correlated with those generated by a standard laboratory extraction method. Short-term storage at room temperature resulted in a slight decline in sIgA concentrations, whereas freezing extracts at -20°C kept sIgA levels stable for at least 3 months. Longer-term storage of >5 months, however, led to a significant decline of sIgA concentrations. Multiple freeze-thaw cycles did not affect sIgA levels. This study, therefore, provides the basis for measuring fecal sIgA in lemurs and possibly other strepsirrhines. When samples are processed properly and stored frozen, and when sIgA analysis can be performed within 3 months upon sample collection, fecal sIgA measurements can become a valuable tool for monitoring aspects of immunity and health in both zoo-housed and wild-living lemurs.

Social bonds increase fitness in a range of mammals. One pathway by which social bonds may increase fitness is by reducing the exposure to physiological stress, i.e. glucocorticoid (GC) hormones, that can be detrimental to health and survival. This is achieved through downregulating hypothalamic-pituitary-adrenal (HPA)-axis activity. Indeed, long-term measures of social (grooming) bonds are often negatively correlated with HPA-axis activity. However, the proximate role of physical touch through allogrooming remains an open question in the sociality-health-fitness debate. Demonstrating the potential anxiolytic benefits of grooming in the wild is hindered by methodological limitations. Here, we match accelerometer-identified grooming in wild female chacma baboons (Papio ursinus) to non-invasive faecal GC metabolite concentrations (fGCs). Consistent with previous work, we found a negative (but statistically non-significant) overall relationship between individual averaged fGCs and grooming rates. However, when time-matching grooming to fGCs, we found that both more giving and receiving grooming were followed by higher fGCs. This upregulation of HPA-axis activity suggests that maintaining social bonds (and its ultimate fitness benefits) may come at a shorter-term physiological cost. This finding sheds new light on a ubiquitous social behaviour typically considered 'relaxing' and suggests that sociopositive contact can trigger physiological stress.

Female fertility signals are found across taxa, and the precision of such signals may be influenced by the relative strength of different sexual selection mechanisms. Among primates, more precise signals may be found in species with stronger direct male-male competition and indirect female mate choice, and less precise signals in species with stronger indirect male-male competition (e.g. sperm competition) and direct female mate choice. We tested this hypothesis in a wild population of Kinda baboons in Zambia, combining data on female signals with reproductive hormones (estrogen and progesterone metabolites) and intra- and inter-cycle fertility. We predicted that Kinda baboons will exhibit less precise fertility signals than other baboon species, as they experience weaker direct and stronger indirect male-male competition. The frequency of copulation calls and proceptive behavior did not vary with hormones or intra- or inter-cycle fertility in almost all models. Sexual swelling size was predicted by the ratio of estrogen to progesterone metabolites, and was largest in the fertile phase, but differences in size across days were small. Additionally, there was variability in the timing of ovulation relative to the day of sexual swelling detumescence across cycles and swelling size did not vary with inter-cycle fertility. Our results suggest that female Kinda baboon sexual swellings are less precise indicators of fertility compared to other baboon species, while signals in other modalities do not reflect variation in intra- and inter-cycle fertility. Female Kinda baboon sexual signals may have evolved as a strategy to reduce male monopolizability, allowing for more female control over reproduction by direct mate choice.

Proximate mechanisms of ‘social ageing’, i.e. shifts in social activity and narrowing of social networks, are understudied. It is proposed that energetic deficiencies (which are often seen in older individuals) may restrict movement and, in turn, sociality, but empirical tests of these intermediary mechanisms are lacking. Here, we study wild chacma baboons ( Papio ursinus ), combining measures of faecal triiodothyronine (fT3), a non-invasive proxy for energy availability, high-resolution GPS data (movement and social proximity) and accelerometry (social grooming durations). Higher (individual mean-centred) fT3 was associated with increased residency time (i.e. remaining in the same area longer), which, in turn, was positively related to social opportunities (i.e. close physical proximity). Individuals with more frequent social opportunities received more grooming, whereas for grooming given, fT3 moderated this effect, suggesting an energetic cost of giving grooming. While our results support the spirit of the energetic deficiencies hypothesis, the directionality of the relationship between energy availability and movement is unexpected and suggests that lower-energy individuals may use strategies to reduce the costs of intermittent locomotion. Thus, future work should consider whether age-related declines in sociality may be a by-product of a strategy to conserve energy. This article is part of the discussion meeting issue ‘Understanding age and society using natural populations’.

In response to environmental and social challenges, animals mount physiological “stress responses” involving elevated glucocorticoid (GC) levels, which may have long-term consequences for health and survival. However, the degree to which social factors drive these physiological responses is likely modulated by a species’ social system, including social organisation, dispersal patterns, and the steepness of the dominance hierarchy, which influence the costs and benefits of social interactions. We examined the relationship between dominance rank, unit size, and faecal GC metabolite (fGC) levels in wild female Guinea baboons (Papio papio). Guinea baboons live in multilevel societies with female-biased dispersal and shallow rank relationships. Units consisting of one primary male and associated females form the core of these societies. We hypothesised that females experience higher competition for male protection or access to food in larger units and that lower-ranking females would bear the costs of this competition. We predicted that membership in a larger unit and lower rank would be associated with higher fGC levels. We collected 251 faecal hormone samples from 14 individuals. A capture event during the sampling period allowed us to validate our method's sensitivity to stress responses. However, we found no evidence of a link between unit size or rank and fGC levels, suggesting that neither incurs a significant physiological cost in this species. These results extend our insights into the physiological correlates of behavioural variation in female primates, expanding our understanding of their adaptability and resilience to social stressors in relatively egalitarian multilevel societies with female-biased dispersal.

For males of gregarious species, dominance status and the strength of affiliative relationships can have major fitness consequences. Social dynamics also impose costs by affecting glucocorticoids, mediators of homeostasis and indicators of the physiological response to challenges and within-group competition. We investigated the relationships between dominance, social bonds, seasonal challenges, and faecal glucocorticoid metabolite (fGC) measures in wild Assamese macaques (Macaca assamensis) at Phu Khieo Wildlife Sanctuary, Thailand, combining behavioural data with 4129 samples from 62 adult males over 15 years. Our previous work on this population suggested that increased competition during the mating season was associated with elevated fGC levels and that, unusually for male primates, lower rank position correlated with higher fGC levels. With a much larger dataset and dynamic measures of sociality, we re-examined these relationships and additionally tested the potentially fGC-attenuating effect of social support. Contrary to our previous study, yet consistent with the majority of work on male primates, dominance rank had a positive relationship with fGC levels, as high status correlated with elevated glucocorticoid measures. fGC levels were increased at the onset of the mating season. We demonstrated an fGC-reducing effect of supportive relationships in males and showed that dynamics in affiliation can correlate with dynamics in physiological responses. Our results suggest that in a system with intermediate contest potential, high dominance status can impose physiological costs on males that may potentially be moderated by social relationships. We highlight the need to consider the dynamics of sociality and competition that influence hormonal processes.

Measurement of the health and disease status of free-ranging primates is often limited by a lack of available biomarkers of immune activation and inflammation that can be applied noninvasively via the measurement of urine or fecal samples. Here, we evaluate the potential usefulness of noninvasive urinary measurements of a number of cytokines, chemokines, and other markers of inflammation and infection. We took advantage of surgery-associated inflammation in seven captive rhesus macaques, collecting urine samples before and after the medical interventions. We measured these urine samples for 33 different markers of inflammation and immune activation that are known to be responsive to inflammation and infection in rhesus macaque blood samples, via the Luminex platform. We also measured all samples for concentrations of the soluble urokinase plasminogen activator receptor (suPAR), which we had validated in a prior study as an effective biomarker of inflammation. Despite urine samples being collected in captivity under ideal conditions (clean, no contamination with feces or soil, frozen quickly), 13/33 biomarkers measured via Luminex were found at concentrations below detection limits in >50% of samples. Of the remaining 20 markers, only 2 showed significant increases in response to surgery-IL18 and MPO (myeloperoxidase). However, suPAR measurements of the same samples show a consistent marked increase in response to surgery that is absent from the patterns of IL18 and MPO measurement. Given that our samples were collected under conditions that are greatly preferable to those usually encountered in the field, urinary cytokine measurements via the Luminex platform seem overall unpromising for primate field studies.

Background During development, elevated levels of maternal glucocorticoids (GCs) can have detrimental effects on offspring morphology, cognition, and behavior as well as physiology and metabolism. Depending on the timing of exposure, such effects may vary in strength or even reverse in direction, may alleviate with age, or may concern more stable and long-term programming of phenotypic traits. Maternal effects on gut bacterial diversity, composition, and function, and the persistence of such effects into adulthood of long-lived model species in the natural habitats remain underexplored.Results In a cross-sectional sample of infant, juvenile, and adult Assamese macaques, the timing of exposure to elevated maternal GCs during ontogeny was associated with the gut bacterial community of the offspring. Specifically, naturally varying maternal GC levels during early but not late gestation or lactation were associated with reduced bacterial richness. The overall effect of maternal GCs during early gestation on the gut bacterial composition and function exacerbated with offspring age and was 10 times stronger than the effect associated with exposure during late prenatal or postnatal periods. Instead, variation in maternal GCs during the late prenatal or postnatal period had less pronounced or less stable statistical effects and therefore a weaker effect on the entire bacterial community composition, particularly in adult individuals. Finally, higher early prenatal GCs were associated with an increase in the relative abundance of several potential pro-inflammatory bacteria and a decrease in the abundance of Bifidobacterium and other anti-inflammatory taxa, an effect that exacerbated with age.Conclusions In primates, the gut microbiota can be shaped by developmental effects with strong timing effects on plasticity and potentially detrimental consequences for adult health. Together with results on other macaque species, this study suggests potential detrimental developmental effects similar to rapid inflammaging, suggesting that prenatal exposure to high maternal GC concentrations is a common cause underlying both phenomena. Our findings await confirmation by metagenomic functional and causal analyses and by longitudinal studies of long-lived, ecologically flexible primates in their natural habitat, including developmental effects that originate before birth.

Thyroid hormones are key modulators of development, as well as mediators of environmental conditions, by regulating developmental processes and metabolism in primates. Hormone measurement in noninvasively collected samples, that is, feces and urine, is a valuable tool for studying the endocrine function of wildlife, and recent studies have demonstrated the feasibility of measuring thyroid hormones in fecal samples of zoo-housed and wild nonhuman primates. Our study aimed to (i) validate the measurement of immunoreactive fecal total triiodothyronine (IF-T3) in wild Assamese macaques (Macaca assamensis) and (ii) to investigate its developmental changes and its response to environmental changes, including stress responses, in immature individuals. Fecal samples and environmental parameters were collected from individuals of three social groups of wild Assamese macaques living at Phu Khieo Wildlife Sanctuary, Northeastern Thailand. Our study confirmed the methodological feasibility and biological validity of measuring IF-T3 in this population. Specifically, the biological validation demonstrated higher IF-T3 levels in immatures compared to adults, and higher levels in females during late gestation compared to the preconception stage. Our analysis of IF-T3 levels in developing immature macaques revealed a significant increase with age. Furthermore, we found a positive association between IF-T3 and immunoreactive fecal glucocorticoid levels, an indicator of the physiological stress response. Neither minimum temperature nor fruit abundance predicted variation in IF-T3 levels in the immatures. Our findings indicate the possibility for differing effects of climatic factors and food availability on thyroid hormone level changes in immature versus adult animals and in wild compared to experimental conditions. Overall, our study provides the basis for further investigations into the role of thyroid hormones in shaping species-specific traits, growth, and overall primate development.

Glucocorticoid and androgen hormones play a prominent role in male reproductive effort. Their production usually increases in non-human primates during mating competition, which may include rivalry for access to receptive females, struggles for high dominance rank, or social pressure on low-ranking individuals. It is generally assumed that glucocorticoids and androgens are associated with mating challenges rather than dominance status, but the involvement of multiple factors makes it difficult to disentangle the two. In this regard, Tonkean macaques provide a suitable model because they are characterized by relaxed dominance and yearround breeding, meaning that there is typically no more than one receptive female in a group, and thus firstranking males can easily monopolize her. We studied two captive groups of Tonkean macaques over an 80month period, recording the reproductive status of females, collecting urine from males and sampling behaviors in both sexes. Male urinary hormone concentrations could be affected by increased competition caused by the mating period, the number of males and the degree of female attractiveness. The highest increases in androgens were recorded in males performing female mate-guarding. Despite the importance of dominance status in determining which males can mate, we found no significant effect of male rank on glucocorticoids and only a marginal effect on androgens during mate-guarding. Both types of hormones were more directly involved in the mating effort of males than in their dominance status. Our results show that their function can be understood in light of the particular competitive needs generated by the species-specific social system.

Ecotourism managers and researchers often assume that apparently habituated primate groups no longer experience adverse consequences of prolonged exposure to tourists or researchers. We examined the effects of tourists and researchers on fecal glucocorticoid metabolite output (FGCM) in three critically endangered, wild crested macaque (Macaca nigra) groups in Tangkoko Nature Reserve, Sulawesi, Indonesia. We assayed FGCM from 456 fecal samples collected from thirty-three adults. Tourists can walk through and among macaque groups freely. Hence, we examined the possible effects of tourists both (1) in the reserve when away and not interacting with the study groups and (2) when they were present within the macaque groups. Generalized Linear Mixed Model (GLMM) analysis indicated that when tourists were present in the forest, but not directly among the macaques, FGCM levels in the macaque tourism groups were higher in months with more tourists. When tourists were among the macaque groups, some female macaques experienced rises and subsequent postexposure decreases in FGCM levels, consistent with predictions for acute stress. Male FGCM levels increased with tourist numbers within the group. Nevertheless, they were not significantly different from levels during undisturbed or postexposure conditions. FGCM responses related to researchers in groups varied by group, sex, and tourist presence. However, the temporal patterning of FGCM responses showed little evidence of chronic stress from tourism at this site.