P-Mail: The Information Highway of Nocturnal, but Not Diurnal or Cathemeral, Strepsirrhines.

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.

Shyer and larger bird species show more reduced fear of humans when living in urban environments.

As the natural habitats of many species are degraded or disappear, there is scope for these species to be established in urban habitats. To ease the establishment and maintenance of urban populations of more species we need to better understand what degree of phenotypical change to expect as different species transition into urban environments. During the first stages of urban colonization, behavioural changes such as an increase in boldness are particularly important. A consistent response in urban populations is to decrease the distance at which individuals flee from an approaching human (flight initiation distance, or FID). Performing a phylogenetic generalized least-squares (PGLS) analysis on 130 avian species, I found that the largest changes in FID between rural and urban populations occur in species that are larger-bodied and naturally shy (higher rural FID), two phenotypic traits that are not normally associated with urban colonizers. More unlikely species may thus be able to colonize urban environments, especially if we design cities in ways that promote such urban colonizations.

A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation.

Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in non-capacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage.

Beyond aggression: Androgen-receptor blockade modulates social interaction in wild meerkats.

In male vertebrates, androgens are inextricably linked to reproduction, social dominance, and aggression, often at the cost of paternal investment or prosociality. Testosterone is invoked to explain rank-related reproductive differences, but its role within a status class, particularly among subordinates, is underappreciated. Recent evidence, especially for monogamous and cooperatively breeding species, suggests broader androgenic mediation of adult social interaction. We explored the actions of androgens in subordinate, male members of a cooperatively breeding species, the meerkat (Suricata suricatta). Although male meerkats show no rank-related testosterone differences, subordinate helpers rarely reproduce. We blocked androgen receptors, in the field, by treating subordinate males with the antiandrogen, flutamide. We monitored androgen concentrations (via baseline serum and time-sequential fecal sampling) and recorded behavior within their groups (via focal observation). Relative to controls, flutamide-treated animals initiated less and received more high-intensity aggression (biting, threatening, feeding competition), engaged in more prosocial behavior (social sniffing, grooming, huddling), and less frequently initiated play or assumed a 'dominant' role during play, revealing significant androgenic effects across a broad range of social behavior. By contrast, guarding or vigilance and measures of olfactory and vocal communication in subordinate males appeared unaffected by flutamide treatment. Thus, androgens in male meerkat helpers are aligned with the traditional trade-off between promoting reproductive and aggressive behavior at a cost to affiliation. Our findings, based on rare endocrine manipulation in wild mammals, show a more pervasive role for androgens in adult social behavior than is often recognized, with possible relevance for understanding tradeoffs in cooperative systems.

Chemical differences between voided and bladder urine in the aye-aye (Daubentonia madagascariensis): implications for olfactory communication studies.

Urine serves a communicative function in many mammalian species. In some species, the signaling function of urine can be enhanced by the addition of chemical compounds from glands along the distal portion of the urogenital tract. Although urine marking is the main mode of chemical communication in many primate species, there has been no study of the contribution of urogenital secretions to the chemical complexity of primate urine. Here, we compared the chemical composition of bladder urine versus voided urine in the aye-aye, Daubentonia madagascariensis, a strepsirrhine primate that relies on urine in intraspecific communication. Both types of urine, collected from each of 11 aye-ayes representing both sexes of varying adult ages, underwent headspace analysis via gas chromatography and mass spectrometry. Although the average number of compounds was similar in bladder and voided urine, 17% of the compounds detected occurred exclusively in voided urine (but only in a subset of individuals). An overall measure of chemical complexity (using a nonmetric multidimensional scaling analysis) showed that both types of urine were chemically different at the individual level. There was no apparent sex or age differences in the chemical components found in aye-aye urine. Nonetheless, the individual dissimilarities between bladder urine and voided urine indicate chemical contributions from structures along the urogenital tract and offer further support for the relevance of urinary communication in the aye-aye.

The "secret" in secretions: methodological considerations in deciphering primate olfactory communication.

Olfactory communication in primates is gaining recognition; however, studies on the production and perception of primate scent signals are still scant. In general, there are five tasks to be accomplished when deciphering the chemical signals contained in excretions and secretions: (1) obtaining the appropriate samples; (2) extracting the target organic compounds from the biological matrix; (3) separating the extracted compounds from one another (by gas chromatography, GC or liquid chromatography, LC); (4) identifying the compounds (by mass spectrometry, MS and associated procedures); and (5) revealing biologically meaningful patterns in the data. Ultimately, because some of the compounds identified in odorants may not be relevant, associated steps in understanding signal function involve verifying the perception or biological activity of putative semiochemicals via (6) behavioral bioassays or (7) receptor response studies. This review will focus on the chemical analyses and behavioral bioassays of volatile, primate scent signals. Throughout, we highlight the potential pitfalls of working with highly complex, chemical matrices and suggest ways for minimizing problems. A recurring theme in this review is that multiple approaches and instrumentation are required to characterize the full range of information contained in the complex mixtures that typify primate or, indeed, many vertebrate olfactory cues. Only by integrating studies of signal production with those verifying signal perception will we better understand the function of olfactory communication.

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone on the induction of spermiation and amplexus in the American toad (Anaxyrus americanus).

BACKGROUND: Captive breeding programs for endangered amphibian species often utilize exogenous hormones for species that are difficult to breed. The purpose of our study was to compare the efficacy of two different hormones at various concentrations on sperm production, quantity and quality over time in order to optimize assisted breeding. METHODS: Male American toads (Anaxyrus americanus) were divided into three separate treatment groups, with animals in each group rotated through different concentrations of luteinizing hormone releasing hormone analog (LHRH; 0.1, 1.0, 4.0 and 32 micrograms/toad), human chorionic gonadotropin (hCG; 50, 100, 200, and 300 IU), or the control over 24 hours. We evaluated the number of males that respond by producing spermic urine, the sperm concentration, percent motility, and quality of forward progression. We also evaluated the effects of hCG and LHRH on reproductive behavior as assessed by amplexus. Data were analyzed using the Generalized Estimating Equations incorporating repeated measures over time and including the main effects of treatment and time, and the treatment by time interaction. RESULTS: The hormone hCG was significantly more effective at stimulating spermiation in male Anaxyrus americanus than LHRH and showed a dose-dependent response in the number of animals producing sperm. At the most effective hCG dose (300 IU), 100% of the male toads produced sperm, compared to only 35% for the best LHRH dose tested (4.0 micrograms). In addition to having a greater number of responders (P < 0.05), the 300 IU hCG treatment group had a much higher average sperm concentration (P < 0.05) than the treatment group receiving 4.0 micrograms LHRH. In contrast, these two treatments did not result in significant differences in sperm motility or quality of forward progressive motility. However, more males went into amplexus when treated with LHRH vs. hCG (90% vs. 75%) by nine hours post-administration. CONCLUSION: There is a clear dichotomy between the two hormones' physiological responses on gamete production and stimulation of amplexus. Understanding how these two hormones influence physiology and reproductive behaviors in amphibians will have direct bearing on establishing similar breeding protocols for endangered species.

Avoidance of interspecific mating in female Syrian hamsters is stronger toward familiar than toward unfamiliar heterospecific males.

Adult Syrian hamster females (Mesocricetus auratus) learn to discriminate against familiar heterospecific males (Turkish hamster, M. brandti). We investigated whether females learn to avoid any heterospecific male after exposure to just one heterospecific male. We predicted that, after being exposed to one heterospecific male, a female would avoid mating not only with that familiar male but also with any unfamiliar heterospecific male. We exposed females to a heterospecific male across a wire-mesh barrier for 8 days and then paired the female with (a) that same heterospecific male or (b) an unfamiliar heterospecific male. Females exhibited lordosis faster and for a longer duration toward the unfamiliar than toward the familiar heterospecific male. However, females were similarly aggressive toward familiar and unfamiliar heterospecific males. Perhaps exposure to stimuli from several heterospecific males (a likely scenario in the wild) would result in females behaving similarly toward familiar and unfamiliar heterospecific males.

Syrian hamster males below an age threshold do not elicit aggression from unfamiliar adult males.

In many species, young males are the dispersers, leaving their natal area after weaning to establish a breeding area of their own. As young males disperse, however, they are bound to encounter unfamiliar adult males with established territories. Such interactions between an adult male and a young male may always be agonistic. Alternatively, there may be an age threshold below which aggression is not elicited and above which the adult male is aggressive toward the juvenile male. To test these two alternative hypotheses, we paired 47 young Syrian hamster (Mesocricetus auratus) males ranging from 24 to 65 days of age with 47 adult male hamsters and measured aggressive and investigatory behavior for 5 min. We observed no aggression by the adult toward young males between 24 and 47 days of age or toward the single male that was 49 days of age. Young males that were 50 days of age or older, however, elicited significant levels of aggression from the adults. These results indicate that in Syrian hamsters, young males are less vulnerable to adult aggression up to an age threshold and are more vulnerable to adult aggression beyond that threshold. This pattern may facilitate the establishment of territories by dispersing young males below that age threshold.

Male mammals respond to a risk of sperm competition conveyed by odours of conspecific males.

Sperm competition occurs when a female copulates with two or more males and the sperm of those males compete within the female's reproductive tract to fertilize her eggs. The frequent occurrence of sperm competition has forced males of many species to develop different strategies to overcome the sperm of competing males. A prevalent strategy is for males to increase their sperm investment (total number of sperm allocated by a male to a particular female) after detecting a risk of sperm competition. It has been shown that the proportion of sperm that one male contributes to the sperm pool of a female is correlated with the proportion of offspring sired by that male. Therefore, by increasing his sperm investment a male may bias a potential sperm competition in his favour. Here we show that male meadow voles, Microtus pennsylvanicus, increase their sperm investment when they mate in the presence of another male's odours. Such an increase in sperm investment does not occur by augmenting the frequency of ejaculations, but by increasing the amount of sperm in a similar number of ejaculations.

Anogenital distance predicts female choice and male potency in prairie voles.

Anogenital distance (AGD) in rodents is a useful indicator of masculinization or feminization due to prenatal hormonal effects. If such cues convey useful information about both 'maleness' and more importantly mate quality, then females may select males based on this cue or other cues related to it. We tested this hypothesis by asking if female prairie voles (Microtus ochrogaster) preferred males with relatively longer AGDs and if AGD correlated with fitness enhancing characteristics such as sperm count, sperm size, and gonad size. Not only did preferred males have significantly longer AGD and larger testes than nonpreferred males, but AGD was directly related to the testes size, seminal vesicle size, and the number of sperm stored. We re-evaluated data collected in semi-natural field enclosures and discovered that males that were members of a pairbond had longer AGD than single males. This later result, taken under semi-natural conditions, was consistent with results we obtained in the laboratory. Taken together these data indicate that AGD serves as a useful cue of male potency and that females preferentially associate with males that demonstrate this masculinized phenotype. Moreover, these data imply that females may select mates based on their potential to effectively fertilize ova, a potentially important trait for a species that forms life-long pairbonds.

Differences in the fatty-acid composition of rodent spermatozoa are associated to levels of sperm competition.

Sperm competition is a prevalent phenomenon that drives the evolution of sperm function. High levels of sperm competition lead to increased metabolism to fuel higher sperm velocities. This enhanced metabolism can result in oxidative damage (including lipid peroxidation) and damage to the membrane. We hypothesized that in those species experiencing high levels of sperm competition there are changes in the fatty-acid composition of the sperm membrane that makes the membrane more resistant to oxidative damage. Given that polyunsaturated fatty acids (PUFAs) are the most prone to lipid peroxidation, we predicted that higher sperm competition leads to a reduction in the proportion of sperm PUFAs. In contrast, we predicted that levels of sperm competition should not affect the proportion of PUFAs in somatic cells. To test these predictions, we quantified the fatty-acid composition of sperm, testis and liver cells in four mouse species (genus Mus) that differ in their levels of sperm competition. Fatty-acid composition in testis and liver cells was not associated to sperm competition levels. However, in sperm cells, as predicted, an increase in sperm competition levels was associated with an increase in the proportion of saturated fatty-acids (the most resistant to lipid peroxidation) and by a concomitant decrease in the proportion of PUFAs. Two particular fatty acids were most responsible for this pattern (arachidonic acid and palmitic acid). Our findings thus indicate that sperm competition has a pervasive influence in the composition of sperm cells that ultimately may have important effects in sperm function.

Interactions with heterospecific males do not affect how female Mesocricetus hamsters respond to conspecific males.

Reproductive interference includes any interspecific interaction that reduces the fitness of one or both species involved. There are several types of reproductive interference, but they normally involve the direct cost of interacting or mating with heterospecifics. An indirect cost of interacting with heterospecific individuals is a consequent reduction in successful interactions with conspecifics. We tested the hypothesis that being aggressive towards a heterospecific individual will diminish sexual responses towards conspecifics in later encounters. We used two species of Mesocricetus hamsters (Syrian and Turkish hamsters), whose interspecific interactions have previously been determined. We exposed or both exposed and paired Syrian hamster females with a conspecific or a heterospecific male. Five minutes later, we paired all females with a conspecific male and measured the latency to lordosis, the duration of lordosis and any incidence of aggression. We found that (1) interactions with heterospecific males did not affect how females responded to conspecific males in later encounters and (2) previous pairing of female subjects with either conspecific or heterospecific males promoted a faster sexual response by females in subsequent interactions with conspecific males. Thus, aggressive interactions of Syrian hamster females with heterospecific males, contrary to our initial hypothesis, had a positive effect on subsequent interactions with conspecific males.

Metabolic rate limits the effect of sperm competition on mammalian spermatogenesis.

Sperm competition leads to increased sperm production in many taxa. This response may result from increases in testes size, changes in testicular architecture or changes in the kinetics of spermatogenesis, but the impact of each one of these processes on sperm production has not been studied in an integrated manner. Furthermore, such response may be limited in species with low mass-specific metabolic rate (MSMR), i.e., large-bodied species, because they cannot process energy and resources efficiently enough both at the organismic and cellular levels. Here we compare 99 mammalian species and show that higher levels of sperm competition correlated with a) higher proportions of seminiferous tubules, b) shorter seminiferous epithelium cycle lengths (SECL) which reduce the time required to produce sperm, and c) higher efficiencies of Sertoli cells (involved in sperm maturation). These responses to sperm competition, in turn, result in higher daily sperm production, more sperm stored in the epididymides, and more sperm in the ejaculate. However, the two processes that require processing resources at faster rates (SECL and efficiency of Sertoli cells) only respond to sperm competition in species with high MSMR. Thus, increases in sperm production with intense sperm competition occur via a complex network of mechanisms, but some are constrained by MSMR.

Asymmetric learning to avoid heterospecific males in Mesocricetus hamsters.

If a female mates with a male of a closely related species, her fitness is likely to decline. Consequently, females may develop behavioral mechanisms to avoid mating with heterospecific males. In some species, one such mechanism is for adult females to learn to discriminate against heterospecific males after exposure to such males. We have previously shown that adult, female Syrian hamsters (Mesocricetus auratus) learn to discriminate against male Turkish hamsters (Mesocricetus brandti) after exposure to a single heterospecific male during 8 days across a wire-mesh barrier. Here we repeated that experiment but this time we exposed female Turkish hamsters to a male Syrian hamster for 8 days and then measured sexual and aggressive behaviors towards that heterospecific male and towards a conspecific male. In contrast to female Syrian hamsters, female Turkish hamsters did not differ in their latency to go into lordosis or in any measure of aggression towards either type of male. Female Turkish hamsters spent less time in lordosis with the heterospecific male, but the percentage of trials in which females copulated with conspecific and heterospecific males did not differ. When comparing females from both species that had been exposed to a heterospecific male for 8days, female Syrian hamsters copulated less and were more aggressive towards the heterospecific male compared to the behavior of female Turkish hamsters. We discuss how this asymmetric response between females of the two species may be due to the much larger geographical range of Turkish hamsters compared to Syrian hamsters.

Re-feeding food-deprived male meadow voles affects the sperm allocation of their rival males.

An individual's nutritional status affects the manner in which same- and opposite-sex conspecifics respond to that individual, which may affect their fitness. Male meadow voles, Microtus pennsylvanicus, increase their sperm allocation if they encounter the scent mark of an unfamiliar male that is not nutritionally challenged. If, however, the scent mark comes from a male that has been food deprived for 24 hours, stud male voles do not increase their sperm allocation. Food deprived males may be viewed as being lower quality and a reduced risk of sperm competition by rival males. We hypothesized that stud males in promiscuous mating systems tailor their sperm allocations depending on whether rival males have been food deprived and then re-fed. We predicted that newly re-fed males will be considered a strong risk of sperm competition because of the potentially high fitness and survival costs associated with food deprivation in males, and that they will cause stud males to increase their sperm allocation. Our results, however, showed that the recovery period from 24 hours of food deprivation was a relatively slow process. It took between 96 hours and 336 hours of re-feeding male scent donors that were food deprived for 24 hours to induce stud males to increase their sperm allocation to levels comparable to when scent donors were not food deprived. Stud male voles may be conserving the amount of sperm allocated until the male scent donors have recovered from food deprivation and subsequent re-feeding.

Eulemur, me lemur: the evolution of scent-signal complexity in a primate clade.

Signal complexity has been linked to social complexity in vocal, but not chemical, communication. To address this gap, we examined the chemical complexity of male and female glandular secretions in eight species of Eulemur. In this diverse clade of macrosmatic primates, species differ by social or mating system and dominance structure. We applied principal component and linear discriminate analyses to data obtained by gas chromatography/mass spectrometry. Beyond the significant effects on chemical signals of gland type, sex, season and species, we found effects of social variables and phylogeny. Notably, female odours were more chemically complex in multimale-multifemale species than pair-bonded species, whereas male odours were more chemically complex in codominant species than female-dominant species. Also, the traditional sexual dimorphism, whereby male signal complexity exceeds that of females, was present in codominant species, but reversed in female-dominant species. Lastly, a positive relationship between the species' pairwise chemical distances and their pairwise phylogenetic distances supported a gradual, but relatively fast mode of signal evolution. We suggest that the comparative method can be a powerful tool in olfactory research, revealing species differences relevant to the understanding of current signal utility and evolutionary processes. In particular, social complexity in lemurs may have selected for olfactory complexity.

Adult female hamsters require long and sustained exposures to heterospecific males to avoid interspecific mating.

Interspecific mating normally decreases female fitness. In many species, females avoid heterospecific males innately or by imprinting on their parents. Alternatively, adult females could learn to discriminate against heterospecific males after exposure to such males. For example, Syrian hamster (Mesocricetus auratus) females learn to discriminate between conspecific males and Turkish hamster (M. brandti) males during adulthood by exposure to males of both species. Adult females not previously exposed to Turkish hamster males will mate similarly with conspecific and heterospecific males. However, in a previous study we showed that exposure to a heterospecific male and a conspecific male for 8 days led to mating avoidance and aggression towards the heterospecific male. Here we conducted two experiments to investigate how much exposure to the heterospecific male was required for females to avoid mating with the heterospecific male (Experiment 1) and how long that avoidance lasted in the absence of continuous exposure to heterospecific stimuli (Experiment 2). Fast and durable learning would indicate the evolution of an efficient avoidance response. In Experiment 1, females were exposed to a heterospecific male for 1, 4 h, 4 or 8 days and then paired with that male. We found more avoidance of interspecific mating after 4 or 8 days of exposure than after 1 or 4 h of exposure. In Experiment 2, females were exposed to a heterospecific male for 8 days and then paired with that male either 10 min later or 8 days later. We found that after an 8-day delay females were highly sexually receptive to the heterospecific male. Additionally, a comparison between the current experiments and a previous study indicates that female Syrian hamsters do not require concurrent exposure to a conspecific male and a heterospecific male to learn to avoid interspecific mating; exposure to a heterospecific male is sufficient.

Fluoxetine does not prevent interspecific mating between two hamster species.

In a recent study we showed that female Syrian hamsters (Mesocricetus auratus) from a laboratory stock readily mated with male Turkish hamsters (M. brandti). We hypothesized that captivity and/or unconscious selection of the most receptive females by researchers or animal caretakers results in heightened female sexual receptivity and reduces the tendency to reject heterospecific males. To test this hypothesis, we decided to decrease female receptivity by injection of fluoxetine, which increases the levels of serotonin in a number of brain loci, including areas involved in mediating sexual behavior, and determine whether such a decrease in receptivity would result in better discrimination of heterospecific males and rejection of such males as mates. We treated estrous female Syrian hamsters with 20mg/kg fluoxetine or vehicle and paired them with both a conspecific and a heterospecific male (Turkish hamster) in two sequential tests. All females showed similar behavior - they exhibited lordosis toward both conspecific and heterospecific males. We did not observe any sign of aggression. Latency to display lordosis and the duration of lordosis were not affected by fluoxetine. Fluoxetine thus did not have an effect on the behavior of females toward conspecific or heterospecific males. Independent of treatment (fluoxetine or vehicle injection), females did display lordosis significantly faster and for a longer duration in the presence of a conspecific male, even though these differences were small.

Adult female hamsters avoid interspecific mating after exposure to heterospecific males.

When females mate with a heterospecific male, they do not usually produce viable offspring. Thus, there is a selective pressure for females to avoid interspecific mating. In many species, females innately avoid heterospecific males; females can also imprint on their parents to avoid later sexual interactions with heterospecific males. However, it was previously unknown whether adult females can learn to discriminate against heterospecific males. We tested the hypothesis that adult females previously unable to avoid interspecific mating learn to avoid such mating after being exposed to heterospecific males. Syrian hamster (Mesocricetus auratus) females not previously exposed to Turkish hamster (Mesocricetus brandti) males can discriminate between odors of conspecific and heterospecific males, but they mate with either type of male. However, when we exposed adult females to both a conspecific male and a heterospecific male through wire-mesh barriers for 8 days, and then paired them sequentially with the two males, females were more receptive to conspecific males and more aggressive to heterospecific males. When females were paired with the heterospecific male first and the conspecific male second, no female was receptive and all were aggressive to heterospecific males. When females were paired with the conspecific male first, only 43% of females were then aggressive toward the heterospecific male. That is, interactions with conspecific males may decrease a female's ability to properly avoid heterospecific males. Our study clearly shows for the first time that females can learn during adulthood to avoid interspecific mating just by being exposed to stimuli from heterospecific males.