Scent Marks Signal Species, Sex, and Reproductive Status in Tamarins (Saguinus spp., Neotropical Primates).

Olfactory communication is an important mediator of social interactions in mammals, thought to provide information about an individual's identity and current social, reproductive, and health status. In comparison with other taxa such as carnivores and rodents, few studies have examined primate olfactory communication. Tamarins (Callitrichidae) conspicuously deposit odorous secretions, produced by specialized scent glands, in their environment. In this study, we combined behavioral and chemical data on captive cotton-top tamarins, Saguinus oedipus, and bearded emperor tamarins, S. imperator subgrisescens, to examine the role of olfactory communication in the advertisement of species, sex, and reproductive status. We observed no difference in scent-marking behavior between species; however, females marked more frequently than males, and reproductive individuals more than non-reproductive ones. In addition, tamarins predominantly used their anogenital gland when scent-marking, followed by the suprapubic gland. We collected swabs of naturally deposited tamarin anogenital scent marks, and analyzed these samples using headspace gas chromatography-mass spectrometry. Despite a limited sample size, we established differences in tamarin anogenital mark chemical composition between species, sex and reproductive status, and identified 41 compounds. The compounds identified, many of which have been reported in previous work on mammalian semiochemistry, form targets for future bioassay studies to identify semiochemicals. Our non-invasive method for collecting deposited scent marks makes it a promising method for the study of olfactory communication in scent-marking animal species, applicable to field settings and for the study of elusive animals.

Female-Based Patterns and Social Function in Avian Chemical Communication.

Much of the growing interest in avian chemical signals has focused on the role of kin recognition or mate attraction, often with an emphasis on males, with uropygial gland secretions perhaps providing information about an individual's identity and quality. Yet, data collected to date suggest sexual dimorphism in uropygial glands and secretions are often emphasized in female, rather than in male birds. That is, when a sexual difference occurs (often during the breeding season only), it is the female that typically exhibits one of three patterns: (1) a larger uropygial gland, (2) a greater abundance of volatile or semi-volatile preen oil compounds and/or (3) greater diversity of preen oil compounds or associated microbes. These patterns fit a majority of birds studied to date (23 of 30 chemically dimorphic species exhibit a female emphasis). Multiple species that do not fit are confounded by a lack of data for seasonal effects or proper quantitative measures of chemical compounds. We propose several social functions for these secretions in female-based patterns, similar to those reported in mammals, but which are largely unstudied in birds. These include: (1) intersexual advertisement of female receptivity or quality, including priming effects on male physiology, (2) intrasexual competition, including scent marking and reproductive suppression or (3) parental behaviors, such as parent-offspring recognition and chemical protection of eggs and nestlings. Revisiting the gaps of chemical studies to quantify the existence of female social chemosignals and any fitness benefit(s) during breeding are potentially fruitful but overlooked areas of future research.

Identification of potential chemosignals in the European water vole Arvicola terrestris.

The water vole Arvicola terrestris is endemic to Europe where its outbreak generates severe economic losses for farmers. Our project aimed at characterising putative chemical signals used by this species, to develop new sustainable methods for population control that could also be used for this species protection in Great Britain. The water vole, as well as other rodents, uses specific urination sites as territorial and sex pheromone markers, still unidentified. Lateral scent glands and urine samples were collected from wild males and females caught in the field, at different periods of the year. Their volatile composition was analysed for each individual and not on pooled samples, revealing a specific profile of flank glands in October and a specific profile of urinary volatiles in July. The urinary protein content appeared more contrasted as males secrete higher levels of a lipocalin than females, whenever the trapping period. We named this protein arvicolin. Male and female liver transcript sequencing did not identify any expression of other odorant-binding protein sequence. This work demonstrates that even in absence of genome, identification of chemical signals from wild animals is possible and could be helpful in strategies of species control and protection.

Are olfactory traits in a pair-bonded primate under sexual selection? An evaluation of sexual dimorphism in Aotus nancymaae.

OBJECTIVES: Sexual selection has seemingly influenced chemical communication in numerous non-human primates, although it is unclear whether it has influenced strictly pair-living and pair-bonded taxa. The physical similarities between male and female owl monkeys suggest that disruptive selection has not played a role in this taxon. However, given their nocturnality, olfactory traits may show differing patterns of sexual selection than visual traits. If sexual selection has influenced chemical communication in owl monkeys, we expect larger scent glands and greater scent-marking in females given the high degree of paternal care, as has been proposed for callitrichines. MATERIALS AND METHODS: We evaluated sex differences in the qualitative and quantitative descriptions of the subcaudal and perianal glandular regions of captive male (n = 39) and female (n = 36) owl monkeys (A. nancymaae), and in the olfactory behaviors performed within breeding pairs (n = 16). RESULTS: Males had larger areas of secretion retained in the hairs covering the subcaudal gland, and females had more and darker secretion than males covering the perianal region. Males inspected the genital region of their partners more frequently than females did, but the sexes did not differ much in other investigative and marking behaviors. DISCUSSION: The observed sex differences and variation in olfactory traits are consistent with the hypothesis that sexual selection has influenced chemical communication in owl monkeys, with males having larger subcaudal glands and spending more time investigating odors. Still, sex differences in monogamous owl monkeys were less extreme than those in other, non-monogamous, taxa.

The pregenital abdomen of Enicocephalomorpha and morphological evidence for different modes of communication at the dawn of heteropteran evolution.

The internal and external anatomy of the posterior metathoracic region, pregenital abdomen, and associated nervous system of the heteropteran infraorder Enicocephalomorpha are thoroughly described, using an array of state-of-the art techniques. Based on morphology, it is hypothesised which modes of communication these insects use. This study is based primarily on an undescribed species of Cocles Bergroth, 1905 (Enicocephalidae) and another undescribed species of Lomagostus Villiers, 1958 (Aenictopecheidae), but additional representatives of the infraorder are also examined. Our results are compared with the literature on other Heteroptera. The metathoracic scent gland system of Enicocephalomorpha uses the same muscles as that of more derived Heteroptera, although the efferent system is different. The presence of a tergal plate and well-developed longitudinal musculature in the families Enicocephalidae and Aenictopecheidae, as well as a sexually dimorphic set of sclerites and membranes that allow an as yet undetermined type of motion, may indicate the presence of vibrational signaling in the infraorder, although experimental confirmation is required. Our findings raise new research questions regarding heteropteran functional morphology and communication.

Seasonal expressions of follicle-stimulating hormone receptor and luteinizing hormone receptor in the scented gland of the male muskrat (Ondatra zibethicus).

Accumulating evidence has shown that follicle-stimulating hormone (FSH) and luteinizing hormone (LH) may influence the functions of nongonadal tissues in addition to their classic target gonads. Our previous studies revealed that the scented glands of male muskrats expressed prolactin receptor, steroidogenic enzymes, and inhibin/activin subunits. To further seek the evidence of the activities of pituitary gonadotropins in scented glands, we investigated the seasonal expression patterns of FSH receptor (FSHR) and LH/choriogonadotropin receptor (LHCGR). The weight and size of scented glands during the breeding season were significantly higher than those during the nonbreeding season. Immunohistochemical studies showed that FSHR was present in the serous cells of scented glands, whereas LHCGR was present in the interstitial cells. The protein and mRNA expression levels of FSHR and LHCGR were significantly higher in the scented glands during the breeding season than those during the nonbreeding season. Importantly, the levels of circulating FSH and LH were remarkably higher during the breeding season. Taken together, these results suggested that gonadotropins may affect the function of muskrat scented gland via the locally expressed receptors in a season-dependent manner.

Seasonal expression of P450arom and estrogen receptors in scented glands of muskrats (Ondatra zibethicus).

Male muskrats have one pair of scented glands that grow and involute annually. To investigate the annual changes in the scented gland, we measured the expressions of aromatase cytochrome P-450 (P450arom) and estrogen receptors (ERs) in the scented glands. P450arom was expressed in glandular cells and epithelial cells in the scented glands during the breeding season, and only in glandular cells during the nonbreeding season. ERα and ERß were also detected in different types of cells in the scented gland during the breeding and nonbreeding seasons. Both mRNA and protein levels of P450arom, ERα, and ERß were higher in the scented glandular tissues during the breeding season than those during the nonbreeding season. In addition, small RNA sequencing showed that the predicted targets of the significantly changed microRNAs might be the genes encoding P450arom and ERs. In conclusion, the seasonal changes in the expression of P450arom and ERs may be involved in the regulation of scented gland functions.

Olfactory perception and behavioral effects of sex pheromone gland components in Helicoverpa armigera and Helicoverpa assulta.

Two sympatric species Helicoverpa armigera and Helicoverpa assulta use (Z)-11-hexadecenal and (Z)-9-hexadecenal as sex pheromone components in reverse ratio. They also share several other pheromone gland components (PGCs). We present a comparative study on the olfactory coding mechanism and behavioral effects of these additional PGCs in pheromone communication of the two species using single sensillum recording, in situ hybridization, calcium imaging, and wind tunnel. We classify antennal sensilla types A, B and C into A, B1, B2, C1, C2 and C3 based on the response profiles, and identify the glomeruli responsible for antagonist detection in both species. The abundance of these sensilla types when compared with the number of OSNs expressing each of six pheromone receptors suggests that HarmOR13 and HassOR13 are expressed in OSNs housed within A type sensilla, HarmOR14b within B and C type sensilla, while HassOR6 and HassOR16 within some of C type sensilla. We find that for H. armigera, (Z)-11-hexadecenol and (Z)-11-hexadecenyl acetate act as behavioral antagonists. For H. assulta, instead, (Z)-11-hexadecenyl acetate acts as an agonist, while (Z)-9-hexadecenol, (Z)-11-hexadecenol and (Z)-9-hexadecenyl acetate are antagonists. The results provide an overall picture of intra- and interspecific olfactory and behavioral responses to all PGCs in two sister species.

Experimental evidence that litter size imposes an oxidative challenge to offspring.

The post-natal environment in which young develop can substantially impact development, adult phenotype and fitness. In wild mice, competition among litter-mates affects development rate and adult behaviour. We manipulated post-natal litter size in a cross-fostering design to investigate the effects of enlarged and reduced litter sizes on sexual signalling, oxidative stress and the links between them. Oxidative stress causes somatic damage that can limit reproductive success and lifespan, and is predicted to mediate investment in life-history traits, including sexual signals. We predicted that litter enlargement would cause an increase in potential oxidative stress, inhibit growth and reduce sexual signalling in male mice. Males reared in enlarged litters were smaller at weaning and, despite rapid growth immediately after weaning, remained smaller at 10 weeks of age than those reared in smaller litters. Females from enlarged litters were consistently smaller throughout post-weaning development and showed no increase in growth rate compared with females from reduced litters. In enlarged litters, protein thiol concentration was lower at weaning in the liver and kidneys, with this trend continuing at 10 weeks of age in the kidneys only. Aconitase enzyme activity was also lower in mice from enlarged litters at weaning and 10 weeks of age in the kidneys. Male mice from enlarged litters scent marked more frequently and had larger preputial glands than those from reduced litters, indicating greater sexual signalling investment irrespective of this increased oxidative challenge. The results of this study are the first to reveal oxidative costs of developmental stress in small mammals.

The Scent Glands of the Neotropical Harvestman Discocyrtus pectnifemur: Morphology, Behavior and Chemistry.

Harvestmen have a pair of scent glands that open through ozopores. The literature suggests a link between the morphology of the ozopore area and the emission of a defensive secretion. A previous study on a species that aggregates in open areas, where individuals are probably more easily spotted by predators, showed that this defensive secretion causes conspecifics to flee. However, it is unknown whether this behavior occurs in species that aggregate in sheltered areas, where prey are harder to find. Herein, we describe the morphology of the ozopore area, the mode of emission of the defensive secretion, and its chemical composition in the harvestman Discocyrtus pectinifemur. We also tested if the defensive secretion is used as an alarm pheromone. We found that D. pectinifemur releases the defensive secretion in different ways, one of them being as a jet. Emission as a jet contrasts with that known for all congeners previously studied, and is in accord with the expected morphology of the ozopore. We found that the defensive secretion of D. pectinifemur does not function as an alarm pheromone. The composition of the defensive secretion, a mixture of quinones, is congruent with those already described for the clade that includes Discocyrtus. Our results support the link between the morphology of the scent glands area and the emission behavior of the defensive secretion, and they suggest that the alarm pheromone function in harvestmen may be dependent on ecological factors.

First records of preorbital gland opening in rare wild barasingha (Rucervus duvaucelii) in social contexts may help to explain this phenomenon in cervids.

The opening of the preorbital gland in deer serves as a visual communication and has been linked to a wide variety of behavioural situations. As we reported recently, all previous long-term studies on preorbital gland opening were carried out on only one species, whereas case reports on six other rarely studied species have shown associations with different behaviours, thus greatly increasing our overall understanding of the real function of this gland in animal visual communication. Here, we report for the first time preorbital gland opening in the barasingha (Rucervus duvaucelii) in social contexts as observed in a wild population in Kanha National Park, India. We observed this behaviour in two different contexts: agonistic and sexual. Moreover, our record of preorbital gland opening during copulation is the first one amongst cervids. Our findings of preorbital gland opening in both contexts in wild barasingha indicate that the gland was opened only when the individual was highly excited. We suggest that preorbital gland opening may be an important behavioural indicator of an individual involved in a serious intraspecific interaction, and thus a useful tool with which to distinguish between playful and serious behaviours, especially in agonistic and sexual situations.

Pre-orbital gland opening during aggressive interactions in rusa deer (Rusa timorensis).

The opening of the preorbital gland in cervids has a visual meaning and is frequently associated with agonistic and/or stress related situations. Apart from in red deer, this behaviour has scarcely been studied and the range of situations when it may occur remains unclear. In this study we report the unusual case of preorbital gland opening in rusa deer, Rusa timorensis, associated to direct aggressive agonistic interaction (biting/kicking) between two adult hinds. This case observed in Tierpark Berlin (Germany) is the first one ever recorded in female-female interactions in cervids. Preorbital gland opening was also studied in 116 social interactions in Plzen Zoo (Czech Republic). Preorbital gland opening by the dominant adult male was twice observed with relation to alert behaviour, which is also rare. In order to contextualise our observations we summarise the current knowledge about the behaviour associated with preorbital gland opening in R. timorensis and in cervids in general.

Are female starlings able to recognize the scent of their offspring?

BACKGROUND: Although there is growing evidence that birds may have individual chemical profiles that can function in several social contexts, offspring recognition based on olfactory cues has never been explored. This ability should be more likely evolved in colonial birds and/or species suffering brood parasitism, in which the risk of being engaged in costly misdirected parental care is high. METHODOLOGY/PRINCIPAL FINDINGS: We performed a choice experiment to examine whether females of the spotless starling, Sturnus unicolor, a species that is colonial, and where a fraction of the population is exposed to intraspecific brood parasitism, can discriminate between the scent of their offspring and that of unrelated nestlings. We also explored whether the development of the uropygial gland secretion may play a role in such olfactory discrimination by performing the choice experiments to females rearing nestlings of two different ages, that is, without and with developed uropygial glands. Results showed that female starlings did not preferentially choose the scent of their offspring, independently of whether the gland of nestlings was developed or not. CONCLUSIONS/SIGNIFICANCE: Our results suggest that female starlings do not have or do not show the ability to distinguish their offspring based on olfaction, at least up to 12-14 days of nestling age. Further research is needed to examine whether odour-based discrimination may function when fledgling starlings leave the nest and the risk of costly misidentification is likely to increase.

Microbes and animal olfactory communication: Where do we go from here?

We know that microbes contribute to the production of odors that some animals use to communicate, but how common is this phenomenon? Recent studies capitalizing on new molecular technologies are uncovering fascinating associations between microbes and odors of wild animals, but causality is difficult to ascertain. Fundamental questions about the nature of these unique host-microbe interactions also remain unanswered. For instance, do microbes benefit from signaling associations with hosts? How does microbial community structure influence signal production? How do hosts regulate microbes in order to generate appropriate signals? Here, we review the current state of knowledge on microbially produced signals in animals and discuss key research foci that can advance our understanding of microbial-based signaling in the animal world.

Neotropical harvestmen (Arachnida, Opiliones) use sexually dimorphic glands to spread chemicals in the environment.

Sexually dimorphic glands have convergently appeared in animals and are often responsible for the production of pheromones. In the suborder Laniatores of the order Opiliones (Arachnida), glands of such type are widespread, but there is not a single paper on how they are used. Using Scanning Electron Microscopy and a behavioral approach, we describe glandular openings and how these glands are used, in the harvestmen Gryne perlata and Gryne coccinelloides (Cosmetidae). Males of these two species have glandular openings on the metatarsi of legs I and on the metatarsi IV. Males were shown rubbing the glands of the metatarsi I against their other legs, whereas glands on the metatarsi IV are gently touched on the substrate or rubbed either against other legs, or against the substrate. Not all behaviors were seen in both species.

Comparative transcriptome analysis of sex pheromone glands of two sympatric lepidopteran congener species.

Two internal apple feeders of Grapholita molesta and Grapholita dimorpha share two major sex pheromone components (stereoisomers) and exhibit a similar circadian rhythm of mating behavior. This study aimed to determine the genetic factors diversifying these two congeners with respect to sex pheromone biosynthetic machinery. Transcriptomes of sex pheromone glands in both species were analyzed with a deep sequencing technology. To find out the gene(s) responsible for the stereoisomer ratios of G. molesta and G. dimorpha, a hypothetic sex pheromone biosynthetic pathway was predicted based on the transcriptomes of their sex pheromone glands. Some orthologs of Δ10 desaturase and FARs in the synthetic pathway were specifically expressed in sex pheromone glands. The relatively high variation in DNA sequence and expression levels between G. molesta and G. dimorpha suggest their crucial roles in generating differential ratios of the sex pheromone stereoisomers in these two sympatric congeners.

Object-horning in goitered gazelle: agonistic or marking behaviour?

We studied object-horning behaviour in goitered gazelles in the natural, arid environment of Kazakhstan over a 6-year period. We found that object-horning was used by adult males mostly as a threat display during territorial conflicts. Therefore object-horning was observed most frequently in territorial single males during the rut in November-December. Object-horning, though, also had a marking effect, with the males' use of this behaviour leaving visible traces that advertized the location of preorbital and urination-defecation scent marks. Therefore, this pattern also was observed linked with preorbital marking and urination-defecation marking behaviours, especially during the rut. Goitered gazelle males chose the most abundant and eatable shrubs for object horning. In contrast to other gazelle species, object-horning in goitered gazelle was observed much more frequently and at the same rate as preorbital and urination-defecation scent markings. This, then, proved a more vigorous and aggressive level of rutting behaviour of the goitered gazelle compared to tropical gazelles, and most likely connected to the short rutting period in the studied species. We concluded, therefore, that object-horning was a manifold phenomenon that played a very important role in goitered gazelle agonistic displays, but without loosing the marking intention of this behaviour.

Comparative study of the pheromone-manufacturing femoral glands in two sympatric species of lacertid lizards (Acanthodactylus).

Femoral glands are holocrine structures that produce compounds used by lizards as pheromones. Few studies have investigated the morphology and ultrastructure of these glands. We chose a closely related species pair from a lizard family having femoral glands in male and female of both species to illustrate comparative morphology and ultrastructure and their implications for the mechanism of secretion dispersal to the environment. We also aimed to test whether the structure and mechanism of secretion production differ between related species. In addition, we sought to gain a better understanding of the holocrine mechanism of secretion. Femoral glands of selected sympatric lacertid species, Acanthodactylus boskianus and A. scutellatus were studied comparatively using light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). SEM revealed both interspecific and sexual variation in the morphology of the glandular pores. The external morphology suggests the mechanism of the secretion deposition where the convex part of pore-carrying scale is probably used to partition the secretory plug. Histology shows the epithelial cells of the gland duct as an extension of the epidermis with its covering keratin. The glandular acini are composed of germinal and secretory cells. The latter undergo four different stages of differentiation, from the beginning of the formation of secretory granules, through the accumulation of these granules, disintegration and formation of the secretory plug, which protrudes externally. The study considers the sequence of holocrine secretion development, and explains in part how such secretions are deposited on the substrate. Sexual differences at the external morphology level were more evident than interspecific differences.

Gender differences in the anatomy of the perineal glands in Guinea pigs and the effect of castration.

Perineal glands in guinea pigs are part of the sebaceous glandular complex. Their secretions are used for scent marking. This is important for social status and can be seen in both sexes and castrated males. Discrepancy exits about the existence of these glands in female guinea pigs and knowledge of the anatomical consequences of castration on the male perineal glands is sparse. To examine these uncertainties related to gender, perineal glands from 13 sexually mature pet guinea pigs were examined macro- and microscopically. Clear gender differences in the anatomy of perineal glands were found, and castrated males showed signs of atrophy and fatty infiltration in the glands. Females do have perineal glands, although smaller than the glands in the male. The glands are typically sebaceous with multiple excretory ducts. A macroscopic unique feature in the males was the clearly evident orifices of a large excretory duct on each side of the slightly everted perineal sac. However, the reason for this gender difference is not clear. In castrated males, the orifices were atrophied and difficult to see. In addition, the sebaceous glands of the hair follicles in the skin folds of the perineal opening were smaller and less abundant in females and castrated males. The changes in castrated males are presumably linked to the hormonal changes and decreased secretion after castration. The dense keratin layer in the perineal sac was thicker in males than in both castrated males and females and could contribute to the concrement formation seen mainly in males.

[Seasonal peculiarities of female hormonal response of females to conspecific chemical signals of the male in two species of the genus Phodopus].

Changes in diestrous female blood plasma progesterone, estradiol, and cortisol in response to odors of urine and midventral gland secretion (MVGS) of conspecific males during different seasons have been studied in the desert hamster (Phodopus roborowskii and Djungarian hamster (Phodopus sungorus). In desert hamster females, a statistically significant (p < or = 0.05) increase in the plasma hormonal level was observed: for progesterone in response to conspecific male MGVS in all seasons except winter, and to conspecific male urine in spring and summer; for cortisol in response to male MVGS and urine in all seasons except spring; and for estradiol in response to male MGVS in autumn. Djungarian hamster females demonstrated a statistically significant increase in the plasma progesterone level in response to conspecific male MVGS and urine in winter and summer, and no response to the estradiol and cortisol level for both odors in all seasons.