Higher social tolerance is associated with more complex facial behavior in macaques.

The social complexity hypothesis for communicative complexity posits that animal societies with more complex social systems require more complex communication systems. We tested the social complexity hypothesis on three macaque species that vary in their degree of social tolerance and complexity. We coded facial behavior in >3000 social interactions across three social contexts (aggressive, submissive, affiliative) in 389 animals, using the Facial Action Coding System for macaques (MaqFACS). We quantified communicative complexity using three measures of uncertainty: entropy, specificity, and prediction error. We found that the relative entropy of facial behavior was higher for the more tolerant crested macaques as compared to the less tolerant Barbary and rhesus macaques across all social contexts, indicating that crested macaques more frequently use a higher diversity of facial behavior. The context specificity of facial behavior was higher in rhesus as compared to Barbary and crested macaques, demonstrating that Barbary and crested macaques used facial behavior more flexibly across different social contexts. Finally, a random forest classifier predicted social context from facial behavior with highest accuracy for rhesus and lowest for crested, indicating there is higher uncertainty and complexity in the facial behavior of crested macaques. Overall, our results support the social complexity hypothesis.

NetFACS: Using network science to understand facial communication systems.

Understanding facial signals in humans and other species is crucial for understanding the evolution, complexity, and function of the face as a communication tool. The Facial Action Coding System (FACS) enables researchers to measure facial movements accurately, but we currently lack tools to reliably analyse data and efficiently communicate results. Network analysis can provide a way to use the information encoded in FACS datasets: by treating individual AUs (the smallest units of facial movements) as nodes in a network and their co-occurrence as connections, we can analyse and visualise differences in the use of combinations of AUs in different conditions. Here, we present 'NetFACS', a statistical package that uses occurrence probabilities and resampling methods to answer questions about the use of AUs, AU combinations, and the facial communication system as a whole in humans and non-human animals. Using highly stereotyped facial signals as an example, we illustrate some of the current functionalities of NetFACS. We show that very few AUs are specific to certain stereotypical contexts; that AUs are not used independently from each other; that graph-level properties of stereotypical signals differ; and that clusters of AUs allow us to reconstruct facial signals, even when blind to the underlying conditions. The flexibility and widespread use of network analysis allows us to move away from studying facial signals as stereotyped expressions, and towards a dynamic and differentiated approach to facial communication.

Comprehensive search filters for retrieving publications on nonhuman primates for literature reviews (filterNHP).

Nonhuman primates (NHPs) are widely studied across many scientific disciplines using a variety of techniques in diverse environments. Due to the wide scope of NHP research, substantial overlap in research topics and questions can occur, whose resulting scientific evidence is synthesized by literature reviews. Identifying all relevant research on a particular topic involving NHPs can be difficult and time consuming. By adopting objective search development techniques from systematic reviews, we developed search filters to detect all scientific publications involving NHPs in PubMed, PsycINFO (via EBSCOhost), and Web of Science. We compared the performance of our comprehensive NHP search filters to search strings typical of a novice database user (i.e., NHP simple search strings) and validated their sensitivity by combining these searches with a topic search of cortisol related studies. For all comparisons, our comprehensive NHP search filters retrieved considerably more scientific publications than the NHP simple search strings. Importantly, our comprehensive NHP search filters are easy to use (text can be copied and pasted into the database search engine) and detect the most recent publications that have yet to be indexed by the bibliographic databases queried. Additionally, we developed filterNHP, an R package and web-based application (https://filterNHP.dpz.eu), for researchers interested in literature searches involving a taxonomic sub-group of NHPs. filterNHP alleviates time necessary for adapting our comprehensive NHP search filters for a particular NHP sub-group by automatizing the creation of these search filters. Altogether, our comprehensive NHP search filters and those for taxonomic sub-groups generated by filterNHP will enable swift and easy retrieval of the available scientific literature involving NHPs, and thereby help enhance the quality of new NHP literature reviews that guide future scientific research (new experiments) and public policy (e.g., on welfare and conservation).

Testosterone and cortisol are negatively associated with ritualized bonding behavior in male macaques.

Neuroendocrine research on the formation of social bonds has primarily focused on the role of nonapeptides. However, steroid hormones often act simultaneously to either inhibit or facilitate bonding. Testosterone is proposed to mediate a trade-off between male mating effort and nurturing behavior; therefore, low levels are predicted during periods of nurturing infant care and social bonding. In species where social bonding and support regulates hypothalamic-pituitary-adrenal axis activity, we also expect glucocorticoid levels to be low during bonding periods. We investigated how immunoreactive urinary testosterone (iuT) and cortisol (iuC) were related to triadic male-infant-male interactions - a ritualized male bonding behavior - as well as infant care in male Barbary macaques (Macaca sylvanus). We collected >3000 h of behavioral observation data during full-day focal animal follows from 14 adult males and quantified iuT and iuC from 650 urine samples. iuT was negatively correlated with rates of triadic interactions within subjects, but positively correlated between subjects. iuC was negatively correlated with triadic interactions both within and between subjects. Time spent caring for infants was positively correlated to both iuT and iuC within subjects, but not between subjects. The observed negative relationship between iuT and triadic interactions within subjects may be beneficial to lower competitive tendencies between adult males and to not inhibit bond formation. However, the positive correlation of iuT with triadic interactions between subjects was unexpected. We speculate that it could be due to a link between triadic interactions and coalition formation. A negative relationship between triadic interactions and iuC could reflect increased bonding and perceived social support as triadic interactions predict future coalition formation in this species, or reflect buffered tensions between males. The positive relationship of iuT and iuC with infant care suggests that the handling of infants may be less nurturing but rather protective or competitive in this species. Measuring steroid hormones in relation to bonding and nurturing can help us interpret behaviors within the ecological contexts that they occur.

Oxytocin increases after affiliative interactions in male Barbary macaques.

Mammals living in stable social groups often mitigate the costs of group living through the formation of social bonds and cooperative relationships. The neuropeptide hormone oxytocin (OT) is proposed to promote both bonding and cooperation although only a limited number of studies have investigated this under natural conditions. Our aim was to assess the role of OT in bonding and cooperation in male Barbary macaques (Macaca sylvanus). First, we tested for an effect of affiliation - grooming and triadic male-infant-male interactions - with bond and non-bond partners on urinary OT levels. Second, we tested whether grooming interactions (and thus increased OT levels) increase a male's general propensity to cooperate in polyadic conflicts. We collected >4000 h of behavioral data on 14 adult males and measured OT levels from 139 urine samples collected after affiliation and non-social control periods. Urinary OT levels were higher after grooming with any partner. By contrast, OT levels after male-infant-male interactions with any partner or with bond partners were not different from controls but were higher after interactions with non-bond partners. Previous grooming did not increase the likelihood of males to support others in conflicts. Collectively, our results support research indicating that OT is involved in the regulation of adult affiliative relationships. However, our male-infant-male interaction results contradict previous studies suggesting that it is affiliation with bond rather than non-bond partners that trigger the release of OT. Alternatively, OT levels were elevated prior to male-infant-male interactions thus facilitating interaction between non-bond partners. The lack of an association of grooming and subsequent support speaks against an OT linked increase in the general propensity to cooperate.

Measuring urinary cortisol and testosterone levels in male Barbary macaques: A comparison of EIA and LC-MS.

The development of methods to quantify hormones from non-invasively collected samples such as urine or feces has facilitated endocrinology research on wild-living animals. To ensure that hormone measurements are biologically meaningful, method validations are strongly recommended for each new species or sample matrix. Our aim was to validate three commonly used enzyme immunoassays (EIA), one for analysis of cortisol and two for analysis of testosterone, to assess adrenocortical and gonadal endocrine activity, respectively, from the urine of male Barbary macaques. We compared EIA and liquid chromatography-mass spectrometry (LC-MS) results to determine if the EIA measurements truly reflect levels of the target hormone and to determine if antibody cross-reactivities with other steroids were potentially confounding results. Furthermore, we conducted a biological validation of testosterone to ensure that both EIA and LC-MS were able to capture physiologically meaningful differences in hormone levels. We found that cortisol measured by EIA correlated strongly with cortisol measured by LC-MS in both adult and immature males, without the need for deconjugation of steroids in the urine. Both testosterone EIAs correlated strongly with LC-MS in adult males, but only if steroids in the urine were deconjugated by enzymatic hydrolysis prior to analysis. However, in immature males, EIA and LC-MS results did not correlate significantly. Further correlation analyses suggest this is likely due to cross-reactivity of the testosterone antibodies with other adrenal steroids such as cortisol, DHEA, and likely others, which are present at much higher concentrations relative to testosterone in immature males. Testosterone levels were significantly higher in adult compared to immature males as measured by LC-MS but not as measured by EIA. Taken together, our results suggest that the testosterone EIAs are suitable to assess gonadal activity in adult but not immature males, and only if a hydrolysis of the urine is conducted prior to analysis.

Correlates of androgens in wild male Barbary macaques: Testing the challenge hypothesis.

Investigating causes and consequences of variation in hormonal expression is a key focus in behavioral ecology. Many studies have explored patterns of secretion of the androgen testosterone in male vertebrates, using the challenge hypothesis (Wingfield, Hegner, Dufty, & Ball, 1990; The American Naturalist, 136(6), 829-846) as a theoretical framework. Rather than the classic association of testosterone with male sexual behavior, this hypothesis predicts that high levels of testosterone are associated with male-male reproductive competition but also inhibit paternal care. The hypothesis was originally developed for birds, and subsequently tested in other vertebrate taxa, including primates. Such studies have explored the link between testosterone and reproductive aggression as well as other measures of mating competition, or between testosterone and aspects of male behavior related to the presence of infants. Very few studies have simultaneously investigated the links between testosterone and male aggression, other aspects of mating competition and infant-related behavior. We tested predictions derived from the challenge hypothesis in wild male Barbary macaques (Macaca sylvanus), a species with marked breeding seasonality and high levels of male-infant affiliation, providing a powerful test of this theoretical framework. Over 11 months, 251 hr of behavioral observations and 296 fecal samples were collected from seven adult males in the Middle Atlas Mountains, Morocco. Fecal androgen levels rose before the onset of the mating season, during a period of rank instability, and were positively related to group mating activity across the mating season. Androgen levels were unrelated to rates of male-male aggression in any period, but higher ranked males had higher levels in both the mating season and in the period of rank instability. Lower androgen levels were associated with increased rates of male-infant grooming during the mating and unstable periods. Our results generally support the challenge hypothesis and highlight the importance of considering individual species' behavioral ecology when testing this framework.