Influence of season and social context on male giant panda (Ailuropoda melanoleuca) vocal behaviour.

Documenting the different social and behavioural contexts that vocalisations are produced in remains an important step towards understanding the functional relevance of specific call types in a given species' vocal repertoire. In this study we investigated whether seasonal differences and the presence or absence of male and female conspecifics influence the production of male giant panda vocal signals. To this end, captive male giant pandas were observed during and outside of the breeding season in three social contexts: only male conspecific neighbours, only female conspecific neighbours, and a context with no neighbours. We found that males were more likely to bleat, chirp, honk and moan during the breeding season, and showed a tendency to growl more outside of the reproductive period. The contextual analysis revealed that bleats were more likely to be produced by males when opposite-sexed conspecifics are in close attendance during the breeding season. Conversely, males were more likely to chirp when neighboured by males than females or no neighbours. In addition, males were more likely to honk in the absence of neighbouring conspecifics during the breeding season, raising the possibility that these calls function to signal location and gain the attention of potential mates. Moans were produced more often when male giant pandas had male than female neighbours during the breeding season, which may reflect mild aggression towards these same-sexed rivals, whereas the production of barks and growls did not vary according to season or the sex of conspecific neighbours. Our findings underscore the importance of male giant panda bleats for coordinating reproduction and promoting contact with potential mating partners in this non-gregarious species, and yield fresh insights into the function of male honks that warrant further investigation. They also provide a basis for comparison with free-ranging giant panda vocal behaviour that could potentially inform conservation efforts.

Coevolution of vocal signal characteristics and hearing sensitivity in forest mammals.

Although signal characteristics and sensory systems are predicted to co-evolve according to environmental constraints, this hypothesis has not been tested for acoustic signalling across a wide range of species, or any mammal sensory modality. Here we use phylogenetic comparative techniques to show that mammal vocal characteristics and hearing sensitivity have co-evolved to utilise higher frequencies in forest environments - opposite to the general prediction that lower frequencies should be favoured in acoustically cluttered habitats. We also reveal an evolutionary trade-off between high frequency hearing sensitivity and the production of calls with high frequency acoustic energy that suggests forest mammals further optimise vocal communication according to their high frequency hearing sensitivity. Our results provide clear evidence of adaptive signal and sensory system coevolution. They also emphasize how constraints imposed by the signalling environment can jointly shape vocal signal structure and auditory systems, potentially driving acoustic diversity and reproductive isolation.

Vocal behaviour predicts mating success in giant pandas.

Surprisingly little is known about how mammal vocal signals are used to achieve behavioural synchrony in the lead up to copulation. The ability to signal short-term fluctuations in arousal levels and behavioural intention is likely to be particularly important for synchronizing mating behaviour in asocial species, which must overcome their natural avoidance and aggressive tendencies to mate. Here, we examined vocal behaviour during breeding encounters in captive giant pandas (Ailuropoda melanoleuca) to gain a greater understanding of how close-range vocal signalling mediates reproduction in this asocial, and conservation-dependent species. Our results revealed that the occurrence of different giant panda vocalizations and acoustic variation within these calls is predictive of successful encounters leading to copulation, as opposed to unsuccessful encounters that do not. In addition, key differences were detected between vocalizations produced during and just prior to copulation. These findings illustrate that vocal exchanges are crucial for achieving behavioural synchrony and signalling intention to mate in giant pandas, and could also provide a valuable tool for breeding programmes, helping conservation managers to assess the likelihood of breeding introductions leading to copulation or potentially injurious failure.

Sound transmission in a bamboo forest and its implications for information transfer in giant panda (Ailuropoda melanoleuca) bleats.

Although mammal vocalisations signal attributes about the caller that are important in a range of contexts, relatively few studies have investigated the transmission of specific types of information encoded in mammal calls. In this study we broadcast and re-recorded giant panda bleats in a bamboo plantation, to assess the stability of individuality and sex differences in these calls over distance, and determine how the acoustic structure of giant panda bleats degrades in this species' typical environment. Our results indicate that vocal recognition of the caller's identity and sex is not likely to be possible when the distance between the vocaliser and receiver exceeds 20 m and 10 m, respectively. Further analysis revealed that the F0 contour of bleats was subject to high structural degradation as it propagated through the bamboo canopy, making the measurement of mean F0 and F0 modulation characteristics highly unreliable at distances exceeding 10 m. The most stable acoustic features of bleats in the bamboo forest environment (lowest % variation) were the upper formants and overall formant spacing. The analysis of amplitude attenuation revealed that the fifth and sixth formant are more prone to decay than the other frequency components of bleats, however, the fifth formant still remained the most prominent and persistent frequency component over distance. Paired with previous studies, these results show that giant panda bleats have the potential to signal the caller's identity at distances of up to 20 m and reliably transmit sex differences up to 10 m from the caller, and suggest that information encoded by F0 modulation in bleats could only be functionally relevant during close-range interactions in this species' natural environment.

The remarkable vocal anatomy of the koala (Phascolarctos cinereus): insights into low-frequency sound production in a marsupial species.

Koalas are characterised by a highly unusual vocal anatomy, with a descended larynx and velar vocal folds, allowing them to produce calls at disproportionately low frequencies. Here we use advanced imaging techniques, histological data, classical macroscopic dissection and behavioural observations to provide the first detailed description and interpretation of male and female koala vocal anatomy. We show that both males and females have an elongated pharynx and soft palate, resulting in a permanently descended larynx. In addition, the hyoid apparatus has a human-like configuration in which paired dorsal, resilient ligaments suspend the hyoid apparatus from the skull, while the ventral parts tightly connect to the descended larynx. We also show that koalas can retract the larynx down into the thoracic inlet, facilitated by a dramatic evolutionary transformation of the ventral neck muscles. First, the usual retractors of the larynx and the hyoid have their origins deep in the thorax. Secondly, three hyoid muscles have lost their connection to the hyoid skeleton. Thirdly, the genioglossus and geniohyoid muscles are greatly increased in length. Finally, the digastric, omohyoid and sternohyoid muscles, connected by a common tendinous intersection, form a guiding channel for the dynamic down-and-up movements of the ventral hyoid parts and the larynx. We suggest that these features evolved to accommodate the low resting position of the larynx and assist in its retraction during call production. We also confirm that the edges of the intra-pharyngeal ostium have specialised to form the novel, extra-laryngeal velar vocal folds, which are much larger than the true intra-laryngeal vocal folds in both sexes, but more developed and specialised for low frequency sound production in males than in females. Our findings illustrate that strong selection pressures on acoustic signalling not only lead to the specialisation of existing vocal organs but can also result in the evolution of novel vocal structures in both sexes.

Function and Evolution of Vibrato-like Frequency Modulation in Mammals.

Why do distantly related mammals like sheep, giant pandas, and fur seals produce bleats that are characterized by vibrato-like fundamental frequency (F0) modulation? To answer this question, we used psychoacoustic tests and comparative analyses to investigate whether this distinctive vocal feature has evolved to improve the perception of formants, key acoustic components of animal calls that encode important information about the caller's size and identity [1]. Psychoacoustic tests on humans confirmed that vibrato-like F0 modulation improves the ability of listeners to detect differences in the formant patterns of synthetic bleat-like stimuli. Subsequent phylogenetically controlled comparative analyses revealed that vibrato-like F0 modulation has evolved independently in six mammalian orders in vocal signals with relatively high F0 and, therefore, low spectral density (i.e., less harmonic overtones). We also found that mammals modulate the vibrato in these calls over greater frequency extents when the number of harmonic overtones per formant is low, suggesting that this is a mechanism to improve formant perception in calls with low spectral density. Our findings constitute the first evidence that formant perception in non-speech sounds is improved by fundamental frequency modulation and provide a mechanism for the convergent evolution of bleat-like calls in mammals. They also indicate that selection pressures for animals to transmit important information encoded by formant frequencies (on size and identity, for example) are likely to have been a key driver in the evolution of mammal vocal diversity.

The evolution of acoustic size exaggeration in terrestrial mammals.

Recent studies have revealed that some mammals possess adaptations that enable them to produce vocal signals with much lower fundamental frequency (F0) and formant frequency spacing (ΔF) than expected for their size. Although these adaptations are assumed to reflect selection pressures for males to lower frequency components and exaggerate body size in reproductive contexts, this hypothesis has not been tested across a broad range of species. Here we show that male terrestrial mammals produce vocal signals with lower ΔF (but not F0) than expected for their size in mating systems with greater sexual size dimorphism. We also reveal that males produce calls with higher than expected F0 and ΔF in species with increased sperm competition. This investigation confirms that sexual selection favours the use of ΔF as an acoustic size exaggerator and supports the notion of an evolutionary trade-off between pre-copulatory signalling displays and sperm production.

Giant Panda Maternal Care: A Test of the Experience Constraint Hypothesis.

The body condition constraint and the experience condition constraint hypotheses have both been proposed to account for differences in reproductive success between multiparous (experienced) and primiparous (first-time) mothers. However, because primiparous mothers are typically characterized by both inferior body condition and lack of experience when compared to multiparous mothers, interpreting experience related differences in maternal care as support for either the body condition constraint hypothesis or the experience constraint hypothesis is extremely difficult. Here, we examined maternal behaviour in captive giant pandas, allowing us to simultaneously control for body condition and provide a rigorous test of the experience constraint hypothesis in this endangered animal. We found that multiparous mothers spent more time engaged in key maternal behaviours (nursing, grooming, and holding cubs) and had significantly less vocal cubs than primiparous mothers. This study provides the first evidence supporting the experience constraint hypothesis in the order Carnivora, and may have utility for captive breeding programs in which it is important to monitor the welfare of this species' highly altricial cubs, whose survival is almost entirely dependent on receiving adequate maternal care during the first few weeks of life.

Female Sexual Preferences Toward Conspecific and Hybrid Male Mating Calls in Two Species of Polygynous Deer, Cervus elaphus and C. nippon.

The behavioral processes at the basis of hybridization and introgression are understudied in terrestrial mammals. We use a unique model to test the role of sexual signals as a reproductive barrier to introgression by investigating behavioral responses to male sexual calls in estrous females of two naturally allopatric but reproductively compatible deer species, red deer and sika deer. Previous studies demonstrated asymmetries in acoustic species discrimination between these species: most but not all female red deer prefer conspecific over sika deer male calls while female sika deer exhibit no preference differences. Here, we extend this examination of acoustic species discrimination to the role of male sexual calls in introgression between parent species and hybrids. Using two-speaker playback experiments, we compared the preference responses of estrous female red and sika deer to male sexual calls from conspecifics versus red × sika hybrids. These playbacks simulate early secondary contact between previously allopatric species after hybridization has occurred. Based on previous conspecific versus heterospecific playbacks, we predicted that most female red deer would prefer conspecific calls while female sika deer would show no difference in their preference behaviors toward conspecific and hybrid calls. However, results show that previous asymmetries did not persist as neither species exhibited more preferences for conspecific over hybrid calls. Thus, vocal behavior is not likely to deter introgression between these species during the early stages of sympatry. On a wider scale, weak discrimination against hybrid sexual signals could substantially contribute to this important evolutionary process in mammals and other taxa.

The acoustic structure of male giant panda bleats varies according to intersexual context.

Although the acoustic structure of mammal vocal signals often varies according to the social context of emission, relatively few mammal studies have examined acoustic variation during intersexual advertisement. In the current study male giant panda bleats were recorded during the breeding season in three behavioural contexts: vocalising alone, during vocal interactions with females outside of peak oestrus, and during vocal interactions with peak-oestrous females. Male bleats produced during vocal interactions with peak-oestrous females were longer in duration and had higher mean fundamental frequency than those produced when males were either involved in a vocal interaction with a female outside of peak oestrus or vocalising alone. In addition, males produced bleats with higher rates of fundamental frequency modulation when they were vocalising alone than when they were interacting with females. These results show that acoustic features of male giant panda bleats have the potential to signal the caller's motivational state, and suggest that males increase the rate of fundamental frequency modulation in bleats when they are alone to maximally broadcast their quality and promote close-range contact with receptive females during the breeding season.

The Acoustic Structure and Information Content of Female Koala Vocal Signals.

Determining the information content of animal vocalisations can give valuable insights into the potential functions of vocal signals. The source-filter theory of vocal production allows researchers to examine the information content of mammal vocalisations by linking variation in acoustic features with variation in relevant physical characteristics of the caller. Here I used a source-filter theory approach to classify female koala vocalisations into different call-types, and determine which acoustic features have the potential to convey important information about the caller to other conspecifics. A two-step cluster analysis classified female calls into bellows, snarls and tonal rejection calls. Additional results revealed that female koala vocalisations differed in their potential to provide information about a given caller's phenotype that may be of importance to receivers. Female snarls did not contain reliable acoustic cues to the caller's identity and age. In contrast, female bellows and tonal rejection calls were individually distinctive, and the tonal rejection calls of older female koalas had consistently lower mean, minimum and maximum fundamental frequency. In addition, female bellows were significantly shorter in duration and had higher fundamental frequency, formant frequencies, and formant frequency spacing than male bellows. These results indicate that female koala vocalisations have the potential to signal the caller's identity, age and sex. I go on to discuss the anatomical basis for these findings, and consider the possible functional relevance of signalling this type of information in the koala's natural habitat.

Chemosensory discrimination of identity and familiarity in koalas.

Despite numerous descriptive accounts of scent marking in marsupials, rigorous experimentation is rare, and relatively little evidence exists to show that conspecifics use chemical signals to distinguish between individuals or different social groups. In this study a series of olfactory discrimination tests sought to determine whether: (1) male koala sternal scent gland secretions are individually distinctive; and (2) male koalas can differentiate between the scent of familiar and unfamiliar individuals. In the first experiment a habituation-discrimination trial demonstrated that male koalas discriminate between the scent gland secretions of different unfamiliar individuals. In a second experiment male koalas spent significantly more time investigating scent from unfamiliar males than familiar males, supporting the hypothesis that they differentiate between conspecifics based on their familiarity. Taken together these results suggest that male koalas are able to discriminate the identity and familiarity of conspecifics using chemical cues, and provide a platform for further studies investigating the functional role of olfactory communication in this species.

Response of red deer stags (Cervus elaphus) to playback of harsh versus common roars.

Red deer stags (Cervus elaphus) give two distinct types of roars during the breeding season, the "common roar" and the "harsh roar." Harsh roars are more frequent during contexts of intense competition, and characterized by a set of features that increase their perceptual salience, suggesting that they signal heightened arousal. While common roars have been shown to encode size information and mediate both male competition and female choice, to our knowledge, the specific function of harsh roars during male competition has not yet been studied. Here, we investigate the hypothesis that the specific structure of male harsh roars signals high arousal to competitors. We contrast the behavioral responses of free ranging, harem-holding stags to the playback of harsh roars from an unfamiliar competitor with their response to the playback of common roars from the same animal. We show that males react less strongly to sequences of harsh roars than to sequences of common roars, possibly because they are reluctant to escalate conflicts with highly motivated and threatening unfamiliar males in the absence of visual information. While future work should investigate the response of stags to harsh roars from familiar opponents, our observations remain consistent with the hypothesis that harsh roars may signal motivation during male competition, and illustrate how intrasexual selection can contribute to the diversification of male vocal signals.

Menstrual cycle phase alters women's sexual preferences for composers of more complex music.

Over 140 years ago Charles Darwin first argued that birdsong and human music, having no clear survival benefit, were obvious candidates for sexual selection. Whereas the first contention is now universally accepted, his theory that music is a product of sexual selection through mate choice has largely been neglected. Here, I provide the first, to my knowledge, empirical support for the sexual selection hypothesis of music evolution by showing that women have sexual preferences during peak conception times for men that are able to create more complex music. Two-alternative forced-choice experiments revealed that woman only preferred composers of more complex music as short-term sexual partners when conception risk was highest. No preferences were displayed when women chose which composer they would prefer as a long-term partner in a committed relationship, and control experiments failed to reveal an effect of conception risk on women's preferences for visual artists. These results suggest that women may acquire genetic benefits for offspring by selecting musicians able to create more complex music as sexual partners, and provide compelling support for Darwin's assertion 'that musical notes and rhythm were first acquired by the male or female progenitors of mankind for the sake of charming the opposite sex'.

Koalas use a novel vocal organ to produce unusually low-pitched mating calls.

During the breeding season, male koalas produce 'bellow' vocalisations that are characterised by a continuous series of inhalation and exhalation sections, and an extremely low fundamental frequency (the main acoustic correlate of perceived pitch) [1]. Remarkably, the fundamental frequency (F0) of bellow inhalation sections averages 27.1 Hz (range: 9.8-61.5 Hz [1]), which is 20 times lower than would be expected for an animal weighing 8 kg [2] and more typical of an animal the size of an elephant (Supplemental figure S1A). Here, we demonstrate that koalas use a novel vocal organ to produce their unusually low-pitched mating calls.

Free-ranging male koalas use size-related variation in formant frequencies to assess rival males.

Although the use of formant frequencies in nonhuman animal vocal communication systems has received considerable recent interest, only a few studies have examined the importance of these acoustic cues to body size during intra-sexual competition between males. Here we used playback experiments to present free-ranging male koalas with re-synthesised bellow vocalisations in which the formants were shifted to simulate either a large or a small adult male. We found that male looking responses did not differ according to the size variant condition played back. In contrast, male koalas produced longer bellows and spent more time bellowing when they were presented with playbacks simulating larger rivals. In addition, males were significantly slower to respond to this class of playback stimuli than they were to bellows simulating small males. Our results indicate that male koalas invest more effort into their vocal responses when they are presented with bellows that have lower formants indicative of larger rivals, but also show that males are slower to engage in vocal exchanges with larger males that represent more dangerous rivals. By demonstrating that male koalas use formants to assess rivals during the breeding season we have provided evidence that male-male competition constitutes an important selection pressure for broadcasting and attending to size-related formant information in this species. Further empirical studies should investigate the extent to which the use of formants during intra-sexual competition is widespread throughout mammals.

Are men better than women at acoustic size judgements?

Formants are important phonetic elements of human speech that are also used by humans and non-human mammals to assess the body size of potential mates and rivals. As a consequence, it has been suggested that formant perception, which is crucial for speech perception, may have evolved through sexual selection. Somewhat surprisingly, though, no previous studies have examined whether sexes differ in their ability to use formants for size evaluation. Here, we investigated whether men and women differ in their ability to use the formant frequency spacing of synthetic vocal stimuli to make auditory size judgements over a wide range of fundamental frequencies (the main determinant of vocal pitch). Our results reveal that men are significantly better than women at comparing the apparent size of stimuli, and that lower pitch improves the ability of both men and women to perform these acoustic size judgements. These findings constitute the first demonstration of a sex difference in formant perception, and lend support to the idea that acoustic size normalization, a crucial prerequisite for speech perception, may have been sexually selected through male competition. We also provide the first evidence that vocalizations with relatively low pitch improve the perception of size-related formant information.

Experimental tests of mate choice in nonhuman mammals: the need for an integrative approach.

Experimental studies of mate choice have normally focused on non-mammal animal species, in which female mating preferences are based on clearly defined male traits. Because mammals are invariably larger and behaviourally more complex, they are less suited to this type of experimentation. Nevertheless, numerous studies on nonhuman mammals have shown that females appear to actively choose their mates. In this Commentary, I review the current literature to reveal that most experimental tests of mate choice in mammals are unable to reveal the actual male phenotypic trait(s) of female preference, which is crucial for identifying male characteristics under sexual selection. In addition, very few studies take into account female oestrous stage, or quantify the fitness benefits to discriminating females. Future work should concentrate on demonstrating female preferences for specific male traits that are shown by genetic paternity analysis to be correlated with male reproductive success, using setups that control for the effects of male and female mating strategies and in which the actual experiments are performed during the female's peak oestrous period.

Do red deer stags (Cervus elaphus) use roar fundamental frequency (F0) to assess rivals?

It is well established that in humans, male voices are disproportionately lower pitched than female voices, and recent studies suggest that this dimorphism in fundamental frequency (F0) results from both intrasexual (male competition) and intersexual (female mate choice) selection for lower pitched voices in men. However, comparative investigations indicate that sexual dimorphism in F0 is not universal in terrestrial mammals. In the highly polygynous and sexually dimorphic Scottish red deer Cervus elaphus scoticus, more successful males give sexually-selected calls (roars) with higher minimum F0s, suggesting that high, rather than low F0s advertise quality in this subspecies. While playback experiments demonstrated that oestrous females prefer higher pitched roars, the potential role of roar F0 in male competition remains untested. Here we examined the response of rutting red deer stags to playbacks of re-synthesized male roars with different median F0s. Our results show that stags' responses (latencies and durations of attention, vocal and approach responses) were not affected by the F0 of the roar. This suggests that intrasexual selection is unlikely to strongly influence the evolution of roar F0 in Scottish red deer stags, and illustrates how the F0 of terrestrial mammal vocal sexual signals may be subject to different selection pressures across species. Further investigations on species characterized by different F0 profiles are needed to provide a comparative background for evolutionary interpretations of sex differences in mammalian vocalizations.

Visualizing sound emission of elephant vocalizations: evidence for two rumble production types.

Recent comparative data reveal that formant frequencies are cues to body size in animals, due to a close relationship between formant frequency spacing, vocal tract length and overall body size. Accordingly, intriguing morphological adaptations to elongate the vocal tract in order to lower formants occur in several species, with the size exaggeration hypothesis being proposed to justify most of these observations. While the elephant trunk is strongly implicated to account for the low formants of elephant rumbles, it is unknown whether elephants emit these vocalizations exclusively through the trunk, or whether the mouth is also involved in rumble production. In this study we used a sound visualization method (an acoustic camera) to record rumbles of five captive African elephants during spatial separation and subsequent bonding situations. Our results showed that the female elephants in our analysis produced two distinct types of rumble vocalizations based on vocal path differences: a nasally- and an orally-emitted rumble. Interestingly, nasal rumbles predominated during contact calling, whereas oral rumbles were mainly produced in bonding situations. In addition, nasal and oral rumbles varied considerably in their acoustic structure. In particular, the values of the first two formants reflected the estimated lengths of the vocal paths, corresponding to a vocal tract length of around 2 meters for nasal, and around 0.7 meters for oral rumbles. These results suggest that African elephants may be switching vocal paths to actively vary vocal tract length (with considerable variation in formants) according to context, and call for further research investigating the function of formant modulation in elephant vocalizations. Furthermore, by confirming the use of the elephant trunk in long distance rumble production, our findings provide an explanation for the extremely low formants in these calls, and may also indicate that formant lowering functions to increase call propagation distances in this species'.