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The gut microbiome is gaining recognition for its role in primate nutrition, but we stand to benefit from microbiome comparisons across diverse hosts and environmental conditions. We compared gut microbiome structure in four lemur species from four phylogenetic lineages, including 9 individual mouse lemurs (Microcebus danfossi), 6 brown lemurs (Eulemur fulvus), 20 sifakas (Propithecus coquereli), and a single sportive lemur (Lepilemur grewcockorum). In northwestern Madagascar, these species are sympatric, but use different feeding strategies to cope with environmental challenges, including relying on tree gums and insects (mouse lemurs), and some vs. significant leaf matter (brown lemurs vs. sifakas and sportive lemurs). From one fecal sample collected per lemur in the dry season in the Anjajavy Forest, we determined gut microbiome diversity, variability, and membership via 16S rRNA sequencing. The lemurs harbored strongly species-specific gut microbiomes. Brown lemurs showed more diverse and generalized consortia; mouse lemurs, sifakas, and the sportive lemur had less diverse consortia with more distinct memberships. Consistent with their fallback foods, mouse lemur microbiomes included taxa putatively associated with gum and insect digestion, whereas those of sifakas and the sportive lemur showed stronger and distinct signatures of leaf fiber and secondary compound metabolism. These results point to feeding strategy, intertwined with host phylogeny, as a driver of gut microbiome composition, but highlight real-time dietary specificity as a contributing driver of microbiome diversity. While illuminating how gut microbiomes facilitate host nutrition on challenging foods, these results help explain how ecologically diverse primates living in sympatry may differentially cope with seasonal or stochastic lean times.

A prominent body of research spanning disciplines has been focused on the potential underlying role for oxytocin in the social signatures of monogamous mating bonds. Behavioral differences between monogamous and non-monogamous vole species, putatively mediated by oxytocinergic function, constitute a key source of support for this mechanism, but it is unclear to what extent this hormone–behavior linkage extends to the primate order. In a preregistered experiment, we test if oxytocin receptor blockade affects affiliative behavior in mixed-sex pairs ofEulemur, a genus of strepsirrhine primate containing both monogamous and non-monogamous species. Inconsistent with past studies in monogamous voles or monkeys, we do not find confirmatory evidence inEulemurthat monogamous pairs affiliate more than non-monogamous pairs, nor that oxytocin receptor blockade of one pair member selectively corresponds to reduced affiliative or scent-marking behavior in monogamous species. We do, however, find exploratory evidence of a pattern not previously investigated: simultaneously blocking oxytocin receptors in both members of a monogamous pair predicts lower rates of affiliative behavior relative to controls. Our study demonstrates the value of non-traditional animal models in challenging generalizations based on model organisms, and of methodological reform in providing a potential path forward for behavioral oxytocin research.

In the decades since female social dominance was first described in strepsirrhine primates, researchers have sought to uncover the proximate and ultimate explanations for its development. In the females of various female-dominant species, androgens have been implicated as regulators of behavior and/or predictors of seasonal fluctuations in aggression (the 'Female Masculinization Hypothesis'). Males, more generally, respond to changing social demands via seasonal fluctuations in androgen-mediated behavior (the 'Challenge Hypothesis'), that may also entail changes in activation of the hypothalamic-pituitary-adrenal axis. Here, we explore if androgens, glucocorticoids, and intersexual behavior fluctuate seasonally in the female-dominant, blue-eyed black lemur (Eulemur flavifrons), with potential consequences for understanding female aggression and male deference. Across two studies conducted during the breeding and nonbreeding seasons, we assessed rates of mixed-sex, dyadic social behavior (aggression and affiliation) and concentrations of fecal glucocorticoid metabolites (Study 1) and serum sex hormones (androstenedione, testosterone, and estradiol; Study 2). Our results align with several predictions inspired by the Female Masculinization and Challenge Hypotheses for intersexual relations: During the breeding season, specifically, both aggression and androstenedione peaked in females, while female-initiated affiliation decreased, potentially to facilitate female resource access and reproductive control. By comparison, all target hormones (androgens, estrogen, and glucocorticoids) peaked in males, with glucocorticoid concentrations potentially increasing in response to the surge in female aggression, and unusually high estrogen concentrations year-round potentially facilitating male deference via male-initiated affiliation. These results suggest complex, seasonally and hormonally mediated behavior in Eulemur flavifrons.

This article is part of a Special Issue (Hormones and Hierarchies). To gain more balanced understanding of sexual selection and mammalian sexual differentiation processes, this review addresses behavioral sex differences and hormonal mediators of intrasexual competition in the meerkat (Suricata suricatta) - a cooperative breeder unusual among vertebrates in its female aggression, degree of reproductive skew, and phenotypic divergence. Focused on the evolution, function, mechanism, and development of female dominance, the male remains a key reference point throughout. Integrated review of endocrine function does not support routine physiological suppression in subordinates of either sex, but instead a ramp up of weight, reproduction, aggression, and sex steroids, particularly androgens, in dominant females. Important and timely questions about female competition are thus addressed by shifting emphasis from mediators of reproductive suppression to mediators of reproductive control, and from organizational and activational roles of androgens in males to their roles in females. Unusually, we ask not only how inequity is maintained, but how dominance is acquired within a lifetime and across generations. Antiandrogens administered in the field to males and pregnant dominant females confirm the importance of androgen-mediated food competition. Moreover, effects of maternal endocrine milieu on offspring development reveal a heritable, androgenic route to female aggression, likely promoting reproductive priority along dominant matrilines. Integrating endocrine measures with long-term behavioral, ecological, morphological, and life-history data on normative and experimental individuals, across life stages and generations, provides better appreciation of the role of naturally circulating androgens in regulating the female phenotype, and sheds new light on the evolution of female dominance, reproductive inequity, and cooperative breeding.

Summary As chemicals that elicit unlearned, functionally specialized, and species-specific responses [ 1 ] or ‘stereotyped behavior’ [ 2 ], pheromones differ from mammalian scent signatures that comprise complex, variable mixtures, convey multiple messages via learned chemical combinations, and elicit generalized responses [ 1 ]. Studying ring-tailed lemur (Lemur catta) behavior and semiochemistry, a recent study by Shirasu, Ito et al. [ 2 ] claimed to have identified “the first sex pheromones in primates.” However, reliance on one male in most chemical procedures and on few females in behavioral procedures constrains statistical analyses and challenges the broad applicability of their findings. Also, the non-independent testing of even fewer signaler–recipient dyads downplays the critical role of learning and memory in primate communication [ 1 ] — an argument that refuted earlier claims of primate pheromones [ 3 , 4 ]. Here, we challenge each of their four highlighted findings and interpretations.

Background Diversity at the Major Histocompatibility Complex (MHC) is critical to health and fitness, such that MHC genotype may predict an individual’s quality or compatibility as a competitor, ally, or mate. Moreover, because MHC products can influence the components of bodily secretions, an individual’s body odors may signal its MHC composition and influence partner identification or mate choice. Here, we investigated MHC-based signaling and recipient sensitivity by testing for odor-gene covariance and behavioral discrimination of MHC diversity and pairwise dissimilarity in a strepsirrhine primate, the ring-tailed lemur (Lemur catta). Methods First, we coupled genotyping of the MHC class II gene, DRB, with gas chromatography-mass spectrometry of genital gland secretions to investigate if functional genetic diversity is signaled by the chemical diversity of lemur scent secretions. We also assessed if the chemical similarity between individuals correlated with their MHC-DRB similarity. Next, we assessed if lemurs discriminated this chemically encoded, genetic information in opposite-sex conspecifics. Results We found that both sexes signaled overall MHC-DRB diversity and pairwise MHC-DRB similarity via genital secretions, but in a sex- and season-dependent manner. Additionally, the sexes discriminated absolute and relative MHC-DRB diversity in the genital odors of opposite-sex conspecifics, suggesting that lemur genital odors function to advertise genetic quality. Conclusions In summary, genital odors of ring-tailed lemurs provide honest information about an individual’s absolute and relative MHC quality. Complementing evidence in humans and Old World monkeys, we suggest that reliance on scent signals to communicate MHC quality may be important across the primate lineage. Electronic supplementary material The online version of this article (10.1186/s12862-019-1486-0) contains supplementary material, which is available to authorized users.

Scent marking is a well-established, but highly variable, mode of communication among strepsirrhine primates. We begin by reviewing this literature, focusing on nocturnal species. Our understanding about the information content of scent signals and the factors driving species diversity remains incomplete, owing to difficulties in acquiring comparative chemical data. We therefore re-examine such a data set, representing the richness and relative abundance of volatile organic compounds (VOCs) in the urine of 12 species (from Galagidae, Lorisidae, Daubentoniidae, Cheirogaleidae, Indriidae, and Lemuridae), to explore differences between nocturnal, diurnal and cathemeral species. As predicted by the variable importance of urine marking across species, the urine of nocturnal strepsirrhines contained the most VOCs and putative semiochemicals, differed significantly in composition from that of diurnal and cathemeral species and showed the strongest species scent “signatures.” Relevant to retracing the evolutionary trajectory of cathemeral strepsirrhines, nocturnal and diurnal species were most differentiated in their VOCs, with cathemeral species being intermediary, but more closely aligned with diurnal species. These data support cathemerality as an ancient expansion of diurnal animals into a nocturnal niche. Consideration of the traits and variables associated with olfactory communication offers a profitable new way for examining species diversity and patterns of evolutionary change.

Background Captive animals, compared to their wild counterparts, generally harbor imbalanced gut microbiota owing, in part, to their altered diets. This imbalance is particularly striking for folivores that fundamentally rely on gut microbiota for digestion, yet rarely receive sufficient dietary fiber in captivity. We examine the critically endangered Coquerel’s sifaka (Propithecus coquereli), an anatomically specialized, rather than facultative, folivore that consumes a seasonal frugo-folivorous diet in the wild, but is provisioned predominantly with seasonal foliage and orchard vegetables in captivity. Using amplicon and metagenomic sequencing applied to fecal samples collected from two wild and one captive population (each comprising multiple groups), we clarify how dietary variation underlies the perturbational effect of captivity on the structure and function of this species’ gut microbiota. Results The gut microbiota of wild sifakas varied by study population, most notably in community evenness and in the abundance of diet-associated microbes from Prevotellaeceae and Lachnospiraceae. Nevertheless, the differences among wild subjects were minor compared to those evident between wild and captive sifakas: Unusually, the consortia of captive sifakas were the most diverse, but lacked representation of endemic Bacteroidetes and metagenomic capacity for essential amino-acid biosynthesis. Instead, they were enriched for complex fiber metabolizers from the Firmicutes phylum, for archaeal methanogens, and for several metabolic pathways putatively linked to plant fiber and secondary compound metabolism. Conclusions The relatively minor differences in gut microbial structure and function between wild sifaka populations likely reflect regional and/or temporal environmental variability, whereas the major differences observed in captive conspecifics, including the loss of endemic microbes, but gain in low-abundance taxa, likely reflect imbalanced or unstable consortia. Indeed, community perturbation may not necessarily entail decreased community diversity. Moreover, signatures of greater fiber degradation indicate that captive sifakas consume a more fibrous diet compared to their wild counterparts. These results do not mirror those typically reported for folivores and herbivores, suggesting that the direction and strength of captivity-induced ‘dysbiosis’ may not be universal across species with similar feeding strategies. We propose that tailored, species-specific dietary interventions in captivity, aimed at better approximating naturally foraged diets, could functionally ‘rewild’ gut microbiota and facilitate successful management of diverse species.

Social status can mediate effects on the immune system, with profound consequences for individual health; nevertheless, most investigators of status-related disparities in free-ranging animals have used faecal parasite burdens to proxy immune function in the males of male-dominant species. We instead use direct measures of innate immune function (complement and natural antibodies) to examine status-related immunocompetence in both sexes of a female-dominant species. The meerkat is a unique model for such a study because it is a cooperatively breeding species in which status-related differences are extreme, evident in reproductive skew, morphology, behaviour, communication and physiology, including that dominant females naturally express the greatest total androgen (androstenedione plus testosterone) concentrations. We found that, relative to subordinates, dominant animals had reduced serum bacteria-killing abilities; also, relative to subordinate females, dominant females had reduced haemolytic complement activities. Irrespective of an individual's sex or social status, androstenedione concentrations (but not body condition, age or reproductive activity) negatively predicted concurrent immunocompetence. Thus, dominant meerkats of both sexes are immunocompromised. Moreover, in female meerkats, androstenedione perhaps acting directly or via local conversion, may exert a double-edged effect of promoting dominance and reproductive success at the cost of increased parasitism and reduced immune function. Given the prominent signalling of dominance in female meerkats, these findings may relate to the immunocompetence handicap hypothesis (ICHH); however, our data would suggest that the endocrine mechanism underlying the ICHH need not be mediated solely by testosterone and might explain trade-offs in females, as well as in males.

Background Inter-population variation in host-associated microbiota reflects differences in the hosts’ environments, but this characterization is typically based on studies comparing few populations. The diversity of natural habitats and captivity conditions occupied by any given host species has not been captured in these comparisons. Moreover, intraspecific variation in gut microbiota, generally attributed to diet, may also stem from differential acquisition of environmental microbes—an understudied mechanism by which host microbiomes are directly shaped by environmental microbes. To more comprehensively characterize gut microbiota in an ecologically flexible host, the ring-tailed lemur (Lemur catta; n = 209), while also investigating the role of environmental acquisition, we used 16S rRNA sequencing of lemur gut and soil microbiota sampled from up to 13 settings, eight in the wilderness of Madagascar and five in captivity in Madagascar or the U.S. Based on matched fecal and soil samples, we used microbial source tracking to examine covariation between the two types of consortia. Results The diversity of lemur gut microbes varied markedly within and between settings. Microbial diversity was not consistently greater in wild than in captive lemurs, indicating that this metric is not necessarily an indicator of host habitat or environmental condition. Variation in microbial composition was inconsistent both with a single, representative gut community for wild conspecifics and with a universal ‘signal of captivity’ that homogenizes the gut consortia of captive animals. Despite the similar, commercial diets of captive lemurs on both continents, lemur gut microbiomes within Madagascar were compositionally most similar, suggesting that non-dietary factors govern some of the variability. In particular, soil microbial communities varied across geographic locations, with the few samples from different continents being the most distinct, and there was significant and context-specific covariation between gut and soil microbiota. Conclusions As one of the broadest, single-species investigations of primate microbiota, our study highlights that gut consortia are sensitive to multiple scales of environmental differences. This finding begs a reevaluation of the simple ‘captive vs. wild’ dichotomy. Beyond the important implications for animal care, health, and conservation, our finding that environmental acquisition may mediate aspects of host-associated consortia further expands the framework for how host-associated and environmental microbes interact across different microbial landscapes.

Contemporary theory that emphasizes the roles of oxytocin and vasopressin in mammalian sociality has been shaped by seminal vole research that revealed interspecific variation in neuroendocrine circuitry by mating system. However, substantial challenges exist in interpreting and translating these rodent findings to other mammalian groups, including humans, making research on nonhuman primates crucial. Both monogamous and non-monogamous species exist within Eulemur, a genus of strepsirrhine primate, offering a rare opportunity to broaden a comparative perspective on oxytocin and vasopressin neurocircuitry with increased evolutionary relevance to humans. We performed oxytocin and arginine vasopressin 1a receptor autoradiography on 12 Eulemur brains from seven closely related species to (1) characterize receptor distributions across the genus, and (2) examine differences between monogamous and non-monogamous species in regions part of putative “pair-bonding circuits”. We find some binding patterns across Eulemur reminiscent of olfactory-guided rodents, but others congruent with more visually oriented anthropoids, consistent with lemurs occupying an ‘intermediary’ evolutionary niche between haplorhine primates and other mammalian groups. We find little evidence of a “pair-bonding circuit” in Eulemur akin to those proposed in previous rodent or primate research. Mapping neuropeptide receptors in these nontraditional species questions existing assumptions and informs proposed evolutionary explanations about the biological bases of monogamy.

Female intrasexual competition can be intense in cooperatively breeding species, with the dominant breeder or matriarch limiting reproduction in subordinates via aggression, eviction or infanticide. In males, these tendencies bidirectionally link to testosterone, but in females, there has been no systematic investigation of androgen-mediated behaviour within and across generations. In 22 wild meerkat (Suricata suricatta) clans, we show that matriarchs 1) express peak androgen concentrations during late gestation, 2) when displaying peak feeding competition, dominance, and evictions, and 3) relative to subordinates, produce offspring that are more aggressive in early development. Late-gestation, antiandrogen treatment of matriarchs 4) reduced their dominance behaviour, was associated with infrequent evictions, decreased social centrality within the clan, 5) increased aggression in cohabiting subordinate dams, and 6) reduced their offspring’s aggression. These effects implicate androgen-mediated aggression in the operation of female sexual selection, and intergenerational transmission of ‘masculinised’ phenotypes in the evolution of meerkat cooperative breeding.

Female intrasexual competition can be intense in cooperatively breeding species, with some dominant breeders (matriarchs) limiting reproduction in subordinates via aggression, eviction or infanticide. In males, such tendencies bidirectionally link to testosterone, but in females, there has been little systematic investigation of androgen-mediated behaviour within and across generations. In 22 clans of wild meerkats (Suricata suricatta), we show that matriarchs 1) express peak androgen concentrations during late gestation, 2) when displaying peak feeding competition, dominance behaviour, and evictions, and 3) relative to subordinates, produce offspring that are more aggressive in early development. Late-gestation antiandrogen treatment of matriarchs 4) specifically reduces dominance behaviour, is associated with infrequent evictions, decreases social centrality within the clan, 5) increases aggression in cohabiting subordinate dams, and 6) reduces offspring aggression. These effects implicate androgen-mediated aggression in the operation of female sexual selection, and intergenerational transmission of masculinised phenotypes in the evolution of meerkat cooperative breeding., ‘In some mammals, matriarchal status can be conferred with androgens. Here, the authors identify effects of androgens that implicate androgen-mediated aggression in female sexual selection in meerkats and intergenerational transmission of masculinised phenotypes in the evolution of meerkat cooperative breeding.’

Antibiotics alter the diversity, structure, and dynamics of host-associated microbial consortia, including via development of antibiotic resistance; however, patterns of recovery from dysbiosis and methods to mitigate negative effects, remain poorly understood. We applied an ecological framework via long-term, integrated study of community structure, across scales, to improve understanding of host-microbe symbiosis during dysbiosis and recovery. We experimentally administered a broad-spectrum antibiotic alone or with subsequent fecal transfaunation to healthy, male ring-tailed lemurs (Lemur catta) and longitudinally tracked the diversity, composition, associations, and resistomes of their gut microbiota. Whereas microbial diversity recovered rapidly in lemurs, antibiotics caused long-term instability in community composition – effects that were attenuated by fecal transfaunation. Antibiotic resistance genes, which were universally present, including in treatment-naïve subjects, increased during and persisted after antibiotic treatment. Long-term, integrated study post antibiotic-induced dysbiosis revealed differential, metric-dependent evidence of recovery, beneficial effects of fecal transfaunation, and negative consequences to lemur resistomes.

The study of human chemical communication benefits from comparative perspectives that relate humans, conceptually and empirically, to other primates. All major primate groups rely on intraspecific chemosignals, but strepsirrhines present the greatest diversity and specialization, providing a rich framework for examining design, delivery and perception. Strepsirrhines actively scent mark, possess a functional vomeronasal organ, investigate scents via olfactory and gustatory means, and are exquisitely sensitive to chemically encoded messages. Variation in delivery, scent mixing and multimodality alters signal detection, longevity and intended audience. Based on an integrative, 19-species review, the main scent source used (excretory versus glandular) differentiates nocturnal from diurnal or cathemeral species, reflecting differing socioecological demands and evolutionary trajectories. Condition-dependent signals reflect immutable (species, sex, identity, genetic diversity, immunity and kinship) and transient (health, social status, reproductive state and breeding history) traits, consistent with socio-reproductive functions. Sex reversals in glandular elaboration, marking rates or chemical richness in female-dominant species implicate sexual selection of olfactory ornaments in both sexes. Whereas some compounds may be endogenously produced and modified (e.g. via hormones), microbial analyses of different odorants support the fermentation hypothesis of bacterial contribution. The intimate contexts of information transfer and varied functions provide important parallels applicable to olfactory communication in humans. This article is part of the Theo Murphy meeting issue ‘Olfactory communication in humans’.

Host-associated microbiomes shape and are shaped by myriad processes that ultimately delineate their symbiotic functions. Whereas a host's stable traits, such as its lineage, relate to gross aspects of its microbiome structure, transient factors, such as its varying physiological state, relate to shorter term, structural variation. Our understanding of these relationships in primates derives principally from anthropoid studies and would benefit from a broader, comparative perspective. We thus examined the vaginal, labial and axillary microbiota of captive, female ring-tailed lemurs (Lemur catta) and Coquerel's sifakas (Propithecus coquereli), across an ovarian cycle, to better understand their relation to stable (e.g. species identity/mating system, body site) and transient (e.g. ovarian hormone concentration, forest access) host features. We used 16S amplicon sequencing to determine microbial composition and enzyme-linked immunosorbent assays to measure serum hormone concentrations. We found marked variation in microbiota diversity and community composition between lemur species and their body sites. Across both host species, microbial diversity was significantly correlated with ovarian hormone concentrations: negatively with progesterone and positively with estradiol. The hosts’ differential forest access related to the diversity of environmental microbes, particularly in axillary microbiomes. Such transient endogenous and exogenous modulators have potential implications for host reproductive health and behavioral ecology.

Across species, diversity at the major histocompatibility complex (MHC) is critical to individual disease resistance and, hence, to population health; however, MHC diversity can be reduced in small, fragmented, or isolated populations. Given the need for comparative studies of functional genetic diversity, we investigated whether MHC diversity differs between populations which are open, that is experiencing gene flow, versus populations which are closed, that is isolated from other populations. Using the endangered ring-tailed lemur (Lemur catta) as a model, we compared two populations under long-term study: a relatively “open,” wild population (n = 180) derived from Beza Mahafaly Special Reserve, Madagascar (2003–2013) and a “closed,” captive population (n = 121) derived from the Duke Lemur Center (DLC, 1980–2013) and from the Indianapolis and Cincinnati Zoos (2012). For all animals, we assessed MHC-DRB diversity and, across populations, we compared the number of unique MHC-DRB alleles and their distributions. Wild individuals possessed more MHC-DRB alleles than did captive individuals, and overall, the wild population had more unique MHC-DRB alleles that were more evenly distributed than did the captive population. Despite management efforts to maintain or increase genetic diversity in the DLC population, MHC diversity remained static from 1980 to 2010. Since 2010, however, captive-breeding efforts resulted in the MHC diversity of offspring increasing to a level commensurate with that found in wild individuals. Therefore, loss of genetic diversity in lemurs, owing to small founder populations or reduced gene flow, can be mitigated by managed breeding efforts. Quantifying MHC diversity within individuals and between populations is the necessary first step to identifying potential improvements to captive management and conservation plans.

If gut microbes influence host behavioral ecology in the short term, over evolutionary time, they could drive host niche differentiation. We explored this possibility by comparing the gut microbiota of Madagascar’s folivorous lemurs from Indriidae and Lepilemuridae. Occurring sympatrically in the eastern rainforest, our four, target species have different dietary specializations, including frugo-folivory (sifakas), young-leaf folivory (indri and woolly lemurs), and mature-leaf folivory (sportive lemurs). We collected fecal samples, from 2013 to 2017, and used amplicon sequencing, metagenomic sequencing, and nuclear magnetic resonance spectroscopy, respectively, to integrate analyses of gut microbiome structure and function with analysis of the colonic metabolome. The lemurs harbored species-specific microbiomes, metagenomes, and metabolomes that were tuned to their dietary specializations: Frugo-folivores had greater microbial and metagenomic diversity, and harbored generalist taxa. Mature-leaf folivores had greater individual microbiome variation, and taxa and metabolites putatively involved in cellulolysis. The consortia even differed between related, young-leaf specialists, with indri prioritizing metabolism of fiber and plant secondary compounds, and woolly lemurs prioritizing amino-acid cycling. Specialized gut microbiota and associated gastrointestinal morphologies enable folivores to variably tolerate resource fluctuation and support nutrient extraction from challenging resources (e.g., by metabolizing plant secondary compounds or recalcitrant fibers), perhaps ultimately facilitating host species’ diversity and specialized feeding ecologies.

Both host phylogenetic placement and feeding strategy influence the structure of the gut microbiome (GMB); however, parsing their relative contributions presents a challenge. To meet this challenge, we compared GMB structure in two genera of lemurs characterized by different dietary specializations, the frugivorous brown lemurs ( Eulemur spp.) and the folivorous sifakas ( Propithecus spp.). These genera sympatrically occupy similar habitats (dry forests and rainforests) and diverged over similar evolutionary timescales. We collected fresh faeces from 12 species (six per host genus), at seven sites across Madagascar, and sequenced the 16S rRNA gene to determine GMB membership, diversity and variability. The lemurs' GMBs clustered predominantly by host genus; nevertheless, within genera, host relatedness did not predict GMB distance between species. The GMBs of brown lemurs had greater evenness and diversity, but were more homogeneous across species, whereas the GMBs of sifakas were differentiated between habitats. Thus, over relatively shallow timescales, environmental factors can override the influence of host phylogenetic placement on GMB phylogenetic composition. Moreover, feeding strategy can underlie the relative strength of host–microbiome coadaptation, with Madagascar's folivores perhaps requiring locally adapted GMBs to facilitate their highly specialized diets.

Research on animal microbiomes is increasingly aimed at determining the evolutionary and ecological factors that govern host-microbiome dynamics, which are invariably intertwined and potentially synergistic. We present three empirical studies related to this topic, each of which relies on the diversity of Malagasy lemurs (representing a total of 19 species) and the comparative approach applied across scales of analysis. In Study 1, we compare gut microbial membership across 14 species in the wild to test the relative importance of host phylogeny and feeding strategy in mediating microbiome structure. Whereas host phylogeny strongly predicted community composition, the same feeding strategies shared by distant relatives did not produce convergent microbial consortia, but rather shaped microbiomes in host lineage-specific ways, particularly in folivores. In Study 2, we compare 14 species of wild and captive folivores, frugivores, and omnivores, to highlight the importance of captive populations for advancing gut microbiome research. We show that the perturbational effect of captivity is mediated by host feeding strategy and can be mitigated, in part, by modified animal management. In Study 3, we examine various scent-gland microbiomes across three species in the wild or captivity and show them to vary by host species, sex, body site, and a proxy of social status. These rare data provide support for the bacterial fermentation hypothesis in olfactory signal production and implicate steroid hormones as mediators of microbial community structure. We conclude by discussing the role of scale in comparative microbial studies, the links between feeding strategy and host-microbiome coadaptation, the underappreciated benefits of captive populations for advancing conservation research, and the need to consider the entirety of an animal's microbiota. Ultimately, these studies will help move the field from exploratory to hypothesis-driven research.

The gut microbiome (GMB) of folivores metabolizes dietary fiber into nutrients, including short-chain fatty acids (SCFAs); however, experiments probing the consequences of foliage quality on host GMBs are lacking. We therefore examined GMB structure and function via amplicon sequencing and Nuclear Magnetic Resonance spectroscopy in 31 captive sifakas (Propithecus coquereli) during dietary manipulations associated with husbandry. Supplementing standard diets with diverse foliage blends, versus with a single plant species, promoted more diverse GMBs, enriched for taxa implicated in plant-fiber metabolism, but depleted in taxa implicated in starch metabolism and bile tolerance. The consumption of diverse blends was associated with greater concentrations of colonic SCFAs. Abundant foliage, via forest access, promoted compositionally distinct and more stable GMBs, but reduced concentrations of SCFAs, possibly reflecting selection of high-quality leaves. In 11 subjects denied forest access, we examined the temporal pace of microbial shifts when supplemental foliage was abruptly switched between diverse blends and single species. The sifaka GMB responded within days, with community diversity and composition closely tracking foliage diversity. By providing experimental evidence that the folivore GMB is sensitive to minor changes in dietary foliage, we reveal the fragility of specialist GMBs, with implications for managing the wellbeing of endangered wildlife.

Diversity at the Major Histocompatibility Complex (MHC) is critical to health and fitness, such that MHC genotype may predict an individual’s quality or compatibility as a competitor, ally, or mate. Moreover, because MHC products can influence the components of bodily secretions, an individual’s body odor may signal its MHC and influence partner identification or mate choice. To investigate MHC-based signaling and recipient sensitivity, we test for odor-gene covariance and behavioral discrimination of MHC diversity and pairwise dissimilarity, under the good genes and good fit paradigms, in a strepsirrhine primate, the ring-tailed lemur (Lemur catta). First, we coupled genotyping with gas chromatography-mass spectrometry to investigate if diversity of the MHC-DRB gene is signaled by the chemical diversity of lemur genital scent gland secretions. We also assessed if the chemical similarity between individuals correlated with their MHC similarity. Next, we assessed if lemurs discriminated this chemically encoded, genetic information in opposite-sex conspecifics. We found that both sexes signaled overall MHC diversity and pairwise MHC similarity via genital secretions, but in a sex- and season-dependent manner. Additionally, both sexes discriminated absolute and relative MHC-DRB diversity in the genital odors of opposite-sex conspecifics, supporting previous findings that lemur genital odors function as advertisement of genetic quality. In this species, genital odors provide honest information about an individual’s absolute and relative MHC quality. Complementing evidence in humans and Old World monkeys, our results suggest that reliance on scent signals to communicate MHC quality may be important across the primate lineage.

Honesty is crucial in animal communication when signallers are conveying information about their condition. Condition dependence implies a cost to signal production; yet, evidence of such cost is scarce. We examined the effects of naturally occurring injury on the quality and salience of olfactory signals in ring-tailed lemurs (Lemur catta). Over a decade, we collected genital secretions from 23 (13 male, 10 female) adults across 34 unique injuries, owing primarily to intra-group fights. Using gas chromatography-mass spectrometry, we tested for differences in the chemical composition of secretions across pre-injury, injury and recovery, in animals that did and did not receive antibiotics. Lemur genital secretions were significantly dampened and altered during injury, with patterns of change varying by sex, season and antibiotics. Using behavioural bioassays (excluding odorants from antibiotic-treated animals), we showed that male ‘recipients’ discriminated injury status based on scent alone, directing more competitive counter marking towards odorants from injured vs. uninjured male ‘signallers.’ That injured animals could not maintain their normal signatures provides rare evidence of the energetic cost to signal production. That conspecifics detected olfactory-encoded ‘weakness’ suggests added behavioural costs: By influencing the likelihood of intra- or inter-sexual conflict, condition-dependent signals could have important implications for socio-reproductive behaviour.

In vertebrates, reproductive endocrine concentrations are strongly differentiated by sex, with androgen biases typifying males and estrogen biases typifying females. These sex differences can be reduced in female-dominant species; however, even the most masculinised of females have less testosterone (T) than do conspecific males. To test if aggressively dominant, female meerkats (Suricata suricatta) may be hormonally masculinised, we measured serum androstenedione (A4), T and estradiol (E2) in both sexes and social classes, during both ‘baseline’ and reproductive events. Relative to resident males, dominant females had greater A4, equivalent T and greater E2 concentrations. Males, whose endocrine values did not vary by social status, experienced increased T during reproductive forays, linking T to sexual behaviour, but not social status. Moreover, substantial E2 concentrations in male meerkats may facilitate their role as helpers. In females, dominance status and pregnancy magnified the unusual concentrations of measured sex steroids. Lastly, faecal androgen metabolites replicated the findings derived from serum, highlighting the female bias in total androgens. Female meerkats are thus strongly hormonally masculinised, possibly via A4’s bioavailability for conversion to T. These raised androgen concentrations may explain female aggressiveness in this species and give dominant breeders a heritable mechanism for their daughters’ competitive edge.

Within animal societies, demographic and social factors, as well as the different reproductive efforts of group members, may influence individual patterns of reproductive success and parasitism. In mammals, such relationships have been studied primarily in male-dominant species. To better understand these linkages in other social systems, we studied a female-dominant cooperative breeder, the meerkat (Suricata suricatta). This species is characterized by intense intrasexual competition and extreme reproductive skew in both sexes. Within adults, we examined heterogeneities in infection by 6 species of endoparasites in relation to host factors (e.g., weight, age, social status, sex, and group size). We explored potential trade-offs between reproduction and parasitism in dominant and subordinate animals of both sexes. Whereas weight and age were predictive of a few parasite taxa, social status or sex predicted parasite species richness and patterns of infection for the majority of parasites examined. Moreover, a significant interaction between sex and status for 2 nematode taxa revealed that dominant females were the most at risk of infection. Lastly, a positive relationship between group size and parasitism was evident in females only. In sum, compared with subordinates, dominant meerkats may experience increased exposure to directly transmitted parasites. Coupled with hormone-mediated immunosuppression, the increased susceptibility of dominant females may reflect energy allocation for preferentially maintaining dominance and breeding status over parasite defense. In species in which female intrasexual competition is intense, this trade-off between reproduction and health may be more pronounced in females than in males.

Meerkats are group-living, insectivorous herpestids in which subordinate members provide extensive care for the dominant female's young. In contrast to some cooperative breeders, subordinate female meerkats are physiologically able to reproduce and occasionally do so successfully; their attempts are more frequently 'suppressed' via eviction or infanticide by the dominant female. Spontaneous abortion and neonatal loss occur with some regularity, further negatively impacting reproductive success. Here, we compared the reproductive outcomes and endocrine profiles, including of serum progesterone (P4), serum estradiol (E2), and fecal glucocorticoid metabolites (fGCm), of dominant and subordinate dams residing within their clans in the Kalahari Desert of South Africa. Our study spanned years of drought, which reduced insect abundance and represented a substantial environmental stressor. Meerkat pregnancies were identified at mid-term and culminated either in spontaneous abortions or full-term deliveries, after which pups were either lost prior to emergence from the natal den (usually within 2days of birth) or emerged at 2-3weeks. Neonatal loss exceeded fetal loss for all females, and contributed to narrowing the status-related disparity in female reproductive output seen during less arid periods. Although E2 concentrations were significantly lower in subordinate than dominant females, they were sufficient to support gestation. Absolute E2 concentrations may owe to androgenic precursors that also attain highest concentrations in dominant dams and may mediate aggression underlying female reproductive skew. Pregnancies terminating in fetal loss were marked by significantly lower P4 concentrations in mid-gestation and modestly lower E2 concentrations overall. Consistently high fGCm concentrations further increased across trimesters, particularly (but not consistently) in subordinates and in aborted pregnancies. Environmental stressors may modulate reproductive outcomes in meerkats through their influence on sex steroids and their effects on intragroup competition. The social and eco-physiological factors affecting intraspecific variation in reproductive output, even in obligate cooperative breeders, may be most apparent during extreme conditions, reflecting the benefits of long-term studies for assessing the impact of climate change.

Background: The gut microbiome (GMB) is the first line of defense against enteric pathogens, which are a leading cause of disease and mortality worldwide. One such pathogen, the protozoan Cryptosporidium, causes a variety of digestive disorders that can be devastating and even lethal. The Coquerel’s sifaka (Propithecus coquereli) – an endangered, folivorous primate endemic to Madagascar – is precariously susceptible to cryptosporidiosis under captive conditions. If left untreated, infection can rapidly advance to morbidity and death. Objective: To gain a richer understanding of the pathophysiology of this pathogen while also improving captive management of endangered species, we examine the impact of cryptosporidiosis on the GMB of a flagship species known to experience a debilitating disease state upon infection. Design: Using 16S sequencing of DNA extracted from sifaka fecal samples, we compared the microbial communities of healthy sifakas to those of infected individuals, across infection and recovery periods. Results: Over the course of infection, we found that the sifaka GMB responds with decreased microbial diversity and increased community dissimilarity. Compared to the GMB of unaffected individuals, as well as during pre-infection and recovery periods, the GMB during active infection was enriched for microbial taxa associated with dysbiosis and rapid transit time. Time to recovery was inversely related to age, with young animals being slowest to recover GMB diversity and full community membership. Antimicrobial treatment during infection caused a significant depletion in GMB diversity. Conclusions: Although individual sifakas show unique trajectories of microbial loss and recolonization in response to infection, recovering sifakas exhibit remarkably consistent patterns, similar to initial community assembly of the GMB in infants. This observation, in particular, provides biological insight into the rules by which the GMB recovers from the disease state. Fecal transfaunation may prove effective in restoring a healthy GMB in animals with specialized diets.

Our understanding of chemical signals in mammals derives principally from studies in which researchers examine signal structure or function within a single species. Despite the unique information to be gained from applying comparable methods across multiple species, comparative studies of chemical signals are extremely limited. Here, we review the available literature on the evolution of chemosignals in male and female strepsirrhine primates (galagos, lorises and lemurs), all of which rely heavily on chemical communication. We draw from a few case studies, but focus our review on two comparative studies. In one, researchers examined the volatile chemical composition of urinary signals across 12 species representing most families within Strepsirrhini, including six 'urine-marking' species and six glandular or 'nonurine-marking' species. In the other, researchers examined the volatile chemical composition of glandular signals in eight Eulemur species differing in social or dominance structure. We highlight five findings. (1) Regardless of the scent source, chemical profiles differ substantially between species, providing reliable species 'scent signatures'. None the less, (2) urine markers express more compounds and have more distinguishable species scent signatures in their urine than do nonurine markers, suggesting specialization of function. Within Eulemur (3) chemical richness is greater in multimale-multifemale species than in pair-bonded species. Moreover, (4) whereas chemical richness of male signals is greater in sexually codominant species, chemical richness of female signals is greater in female-dominant species. Together, the findings from both comparative studies, coupled with case studies, suggest that signal richness is linked to some aspect of the focal species' socioecology. Lastly, (5) regardless of the scent source, strepsirrhine chemosignals evolve gradually over time, but at fast rates and homogeneously within different lineages. Comparative studies reveal patterns that cannot be detected from the single-species approach and are therefore critical for providing new insight into the function and evolution of olfactory signals.

Social complexity, often estimated by group size, is seen as driving the complexity of vocal signals, but its relation to olfactory signals, which arguably arose to function in nonsocial realms, remains underappreciated. That olfactory signals also may mediate within-group interaction, vary with social complexity and promote social cohesion underscores a potentially crucial link with sociality. To examine that link, we integrated chemical and behavioural analyses to ask whether olfactory signals facilitate reproductive coordination in a strepsirrhine primate, the Coquerel's sifaka, Propithecus coquereli. Belonging to a clade comprising primarily solitary, nocturnal species, the diurnal, group-living sifaka represents an interesting test case. Convergent with diurnal, group-living lemurids, sifakas expressed chemically rich scent signals, consistent with the social complexity hypothesis for communication. These signals minimally encoded the sex of the signaller and varied with female reproductive state. Likewise, sex and female fertility were reflected in within-group scent investigation, scent marking and overmarking. We further asked whether, within breeding pairs, the stability or quality of the pair's bond influences the composition of glandular signals and patterns of investigatory or scent-marking behaviour. Indeed, reproductively successful pairs tended to show greater similarity in their scent signals than did reproductively unsuccessful pairs, potentially through chemical convergence. Moreover, scent marking was temporally coordinated within breeding pairs and was influenced by past reproductive success. That olfactory signalling reflects social bondedness or reproductive history lends support to recent suggestions that the quality of relationships may be a more valuable proxy than group size for estimating social complexity. We suggest that olfactory signalling in sifakas is more complex than previously recognized and, as in other socially integrated species, can be a crucial mechanism for promoting group cohesion and maintaining social bonds. Thus, the evolution of sociality may well be reflected in the complexity of olfactory signalling.

The contribution of bacterial fermentation to the production of vertebrate scent signals has long been suspected, but there is still relatively little information about the factors driving variation in microbial composition in animal scent secretions. Our study subject, the meerkat (Suricata suricatta), is a social mongoose that lives in territorial, family groups and relies heavily on scent for social communication. Unusually in mammalian research, extensive life-history data exist for multiple groups inhabiting the same ecosystem, allowing for a study of both individual variation and group differences in the host’s microbial communities. Using a culture-independent sampling technique, we explored the relationship between a signaler’s sex, age/dominance, genotype or group membership, and the microbiota of its anal scent secretions. We found differences in the microbiota of males and females, but only after the animals had reached sexual maturity. Although bacterial communities in meerkat scent secretions were not more similar between kin than between nonkin, they were more similar between members of the same group than between members of different groups. Collectively, these results are consistent with a potential role for reproductive hormones in determining a host’s bacterial assemblages, as well as an influence of sociality (such as intragroup allo-marking behavior) and/or microhabitat in the acquisition of bacterial assemblages. This study provides a key starting point for understanding the role of microbes in the variation of scent composition in mammals.

This review/research paper summarizes data on development of the external genitalia of the spotted hyena, a fascinating mammal noted for extreme masculinization of the female external genitalia. The female spotted hyena is the only extant mammal that mates and gives birth through a pendulous penis-like clitoris. Our studies indicate that early formation of the phallus in both males and females is independent of androgens; indeed the phallus forms before the fetal testes or ovaries are capable of synthesizing androgens. Likewise, pre- and postnatal growth in length of the penis and clitoris is minimally affected by “androgen status”. Nonetheless, several internal morphologies, as well as external surface features of the phallus, are androgen-dependent and thus account for dimorphism between the penis and clitoris. Finally, estrogens play a critical role in penile and clitoral development, specifying the position of the urethral orifice, determining elasticity of the urethral meatus, and facilitating epithelial-epithelial fusion events required for proper formation of the distal urethra/urogenital sinus and prepuce. Accordingly, prenatal inhibition of estrogen synthesis via administration of letrozole (an aromatase inhibitor) leads to malformations of the glans as well as the prepuce (hypospadias). The effects of prenatal androgens, anti-androgens and impaired estrogen synthesis correlated with the tissue expression of androgen and estrogen receptors.

The role of androgens in shaping “masculine” traits in males is a core focus in behavioral endocrinology, but relatively little is known about an androgenic role in female aggression and social dominance. In mammalian models of female dominance, including the ring-tailed lemur (Lemur catta), links to androgens in adulthood are variable. We studied the development of ring-tailed lemurs to address the behavioral basis and ontogenetic mechanisms of female dominance. We measured behavior and serum androgen concentrations in 24 lemurs (8 males, 16 females) from infancy to early adulthood, and assessed their ‘prenatal’ androgen milieu using serum samples obtained from their mothers during gestation. Because logistical constraints limited the frequency of infant blood sampling, we accounted for asynchrony between behavioral and postnatal hormone measurements via imputation procedures. Imputation was unnecessary for prenatal hormone measurements. The typical sex difference in androgen concentrations in young lemurs was consistent with adult conspecifics and most other mammals; however, we found no significant sex differences in rough-and-tumble play. Female (but not male) aggression increased beginning at approximately 15 months, coincident with female puberty. In our analyses relating sexually differentiated behavior to androgens, we found no relationship with activational hormones, but several significant relationships with organizational hormones. Notably, associations of prenatal androstenedione and testosterone with behavior were differentiated, both by offspring sex and by type of behavior within offspring sexes. We discuss the importance of considering (1) missing data in behavioral endocrinology research, and (2) organizational androgens other than testosterone in studies of female dominance.

Compared to other modes of communication, chemical signaling between conspecifics generally has been overlooked in Old World primates, despite the presence in this group of secretory glands and scent-marking behavior, as well as the confirmed production and perception of olfactory signals. In other mammalian species, flehmen is a behavior thought to transport primarily nonvolatile, aqueous-soluble odorants via specialized ducts to the vomeronasal organ (VNO). By contrast, Old World primates are traditionally thought to lack a functional VNO, relying instead on the main olfactory system to process volatile odorants from their environment. Here, in the mandrill (Mandrillus sphinx), we document unusual morphological and behavioral traits that typically are associated with the uptake of conspecific chemical cues for processing by an accessory olfactory system. Notably, we confirmed that both sexes possess open nasopalatine ducts and, in response to the presentation of conspecific odorants, we found that both sexes showed stereotyped behavior consistent with the flehmen response. If, as in other species, flehmen in the mandrill serves to mediate social or reproductive information, we expected its occurrence to vary with characteristics of either the signaler or receiver. Flehmen, particularly in a given male, occurred most often in response to odorants derived from male, as opposed to female, conspecifics. Moreover, odorants derived during the breeding season elicited more flehmen responses than did odorants collected during the birthing season. Lastly, odorants from reproductively cycling females also elicited more responses than did odorants from contracepted females. Although confirming a link between the nasopalatine ducts, flehmen behavior, and olfactory processing in mandrills would require further study, our observations provide new information to suggest anatomical variability within Old World primates, calling further attention to the underappreciated role of chemical communication in this lineage.