Patterns and correlates of potential range shifts of bat species in China in the context of climate change.

Climate change may diminish biodiversity; thus, it is urgent to predict how species' ranges may shift in the future by integrating multiple factors involving more taxa. Bats are particularly sensitive to climate change due to their high surface-to-volume ratio. However, few studies have considered geographic variables associated with roost availability and even fewer have linked the distributions of bats to their thermoregulation and energy regulation traits. We used species distribution models to predict the potential distributions of 12 bat species in China under current and future greenhouse gas emission scenarios (SSP1-2.6 and SSP5-8.5) and examined factors that could affect species' range shifts, including climatic, geographic, habitat, and human activity variables and wing surface-to-mass ratio (S-MR). The results suggest that Ia io, Rhinolophus ferrumequinum, and Rhinolophus rex should be given the highest priority for conservation in future climate conservation strategies. Most species were predicted to move northward, except for I. io and R. rex, which moved southward. Temperature seasonality, distance to forest, and distance to karst or cave were the main environmental factors affecting the potential distributions of bats. We found significant relationships between S-MR and geographic distribution, current potential distribution, and future potential distribution in the 2050s. Our work highlights the importance of analyzing range shifts of species with multifactorial approaches, especially for species traits related to thermoregulation and energy regulation, to provide targeted conservation strategies.

Patrones y correlaciones de los cambios potenciales en la distribución de las especies de murciélago de China en el contexto del cambio climático Resumen El cambio climático puede disminuir la biodiversidad, por lo que es urgente pronosticar cómo puede cambiar en el futuro la distribución de las especies mediante la integración de múltiples factores que involucren a más taxones. Los murciélagos son particularmente sensibles al cambio climático debido a que tienen una gran proporción superficie­volumen. Sin embargo, hay pocos estudios que han considerado las variables asociadas con la disponibilidad de nidos y son todavía menos los que han conectado la distribución de los murciélagos con sus rasgos de termorregulación y regulación de energía. Usamos modelos de distribución de especies para pronosticar la distribución potencial de doce especies de murciélago en China bajo escenarios actuales y futuros de emisión de gases de efecto invernadero (SSP1­2.6 y SSP5­8.5) y analizamos los factores que podrían afectar el cambio en la distribución de las especies, incluyendo las variables climáticas, geográficas, de hábitat y de actividad humana y la proporción entre la superficie del ala y la masa (P S­M). Los resultados sugieren que Ia io, Rhinolophus ferrumequinum y R. rex deberían ser la mayor prioridad de conservación para las estrategias de conservación climáticas en el futuro. Pronosticamos que la mayoría de las especies se desplazarían al norte, a excepción de I. io y R. rex, que se desplazarían hacia el sur. Los principales factores que afectaron la distribución potencial de los murciélagos fueron la estacionalidad de la temperatura, la distancia al bosque y la distancia a la cueva o al karst. Encontramos una relación significativa entre la P S­M y la distribución geográfica, la distribución potencial actual y la distribución potencial para la década de 2050. Nuestra investigación destaca la importancia del análisis de los cambios de distribución de las especies con enfoques multifactoriales, especialmente para los rasgos de especie relacionados con la termorregulación y la regulación de energía, para proporcionar estrategias de conservación focalizadas.

A chromosome-level genome assembly of an avivorous bat species (Nyctalus aviator).

Currently, three carnivorous bat species, namely Ia io, Nyctalus lasiopterus, and Nyctalus aviator, are known to actively prey on seasonal migratory birds (hereinafter referred to as "avivorous bats"). However, the absence of reference genomes impedes a thorough comprehension of the molecular adaptations of avivorous bat species. Herein, we present the high-quality chromosome-scale reference genome of N. aviator based on PacBio subreads, DNBSEQ short-reads and Hi-C sequencing data. The genome assembly size of N. aviator is 1.77 Gb, with a scaffold N50 of 102 Mb, of which 99.8% assembly was anchored into 21 pseudo-chromosomes. After masking 635.1 Mb repetitive sequences, a total of 19,412 protein-coding genes were identified, of which 99.3% were functionally annotated. The genome assembly and gene prediction reached 96.1% and 96.1% completeness of Benchmarking Universal Single-Copy Orthologs (BUSCO), respectively. This chromosome-level reference genome of N. aviator fills a gap in the existing information on the genomes of carnivorous bats, especially avivorous ones, and will be valuable for mechanism of adaptations to dietary niche expansion in bat species.

The Consumption and Diversity Variation Responses of Agricultural Pests and Their Dietary Niche Differentiation in Insectivorous Bats.

Insectivorous bats are generalist predators and can flexibly respond to fluctuations in the distribution and abundance of insect prey. To better understand the effects of bats on arthropod pests, the types of pests eaten by bats and the response of bats to insect prey need to be determined. In this study, we performed DNA metabarcoding to examine prey composition and pest diversity in the diets of four insectivorous species of bats (Hipposideros armiger, Taphozous melanopogon, Aselliscus stoliczkanus, and Miniopterus fuliginosus). We evaluated the correlation between bat activity and insect resources and assessed dietary niche similarity and niche breadth among species and factors that influence prey consumption in bats. We found that the diets of these bats included arthropods from 23 orders and 200 families, dominated by Lepidoptera, Coleoptera, and Diptera. The proportion of agricultural pests in the diet of each of the four species of bats exceeded 40% and comprised 713 agricultural pests, including those that caused severe economic losses. Bats responded to the availability of insects. For example, a higher abundance of insects, especially Lepidoptera, and a higher insect diversity led to an increase in the duration of bat activity. In areas with more abundant insects, the number of bat passes also increased. The dietary composition, diversity, and niches differed among species and were particularly significant between H. armiger and T. melanopogon; the dietary niche width was the greatest in A. stoliczkanus and the narrowest in H. armiger. The diet of bats was correlated with their morphological and echolocation traits. Larger bats preyed more on insects in the order Coleoptera, whereas the proportion of bats consuming insects in the order Lepidoptera increased as the body size decreased. Bats that emitted echolocation calls with a high peak frequency and duration preyed more on insects in the order Mantodea. Our results suggest that dietary niche differentiation promotes the coexistence of different bat species and increases the ability of bats to consume insect prey and agricultural pests. Our findings provide greater insights into the role of bats that prey on agricultural pests and highlight the importance of combining bat conservation with integrated pest management.

Resting posture drives the evolution of agonistic displays in bats.

Agonistic displays are one of the most diverse social behaviors that have important functions in animal's life history. However, their origin and driving factors have largely been unexplored. Here, we evaluated agonistic displays of 71 bat species across 10 families and classified these displays into two categories: (a) boxing displays where a bat attacks its opponent with its wrist and thumb and (b) pushing displays where a bat uses its head or body to hit a rival. We estimated the strength of the phylogenetic signal of the agonistic displays, revealed their origin, and tested the potential evolutionary relationships between agonistic behaviors and body size or resting posture (free hanging vs. contact hanging where the bat is in contact with some surface). We found that agonistic displays were phylogenetically conserved and that boxing displays are the ancestral state. Moreover, we found that bats with a free-hanging resting posture were more likely to exhibit boxing displays than pushing displays. In addition, bats with longer forearms do not have a higher propensity for boxing displays. This study expands our limited knowledge of the evolution of agonistic displays and highlights the importance of resting posture as a driving force in the diversity of agonistic displays.

Protocol for simulating foraging behavior strategy and dual-choice predatory sensory cue experiments in aerial-hawking bats.

The nocturnal and cryptic nature of bats makes it difficult to determine their foraging behavior and predatory sensory cues. Here, we present a protocol for determining the behavioral strategies and sensory cues of bat predation on airborne prey. We describe steps for indoor foraging behavior simulation, dual-choice acoustic playback, and visual presentation experiments. This protocol was used to study the avivorous bat, Ia io, but it can be adapted for studying other aerial-hawking bats and even other taxa. For complete details on the use and execution of this protocol, please refer to Gong et al. (2022).1.

Do nocturnal birds use acoustic and visual cues to avoid predation by bats?

Anti-predation strategies are critical to animal survival and are fundamental to deciphering predator-prey interactions. As an important defense strategy, sensory predator detection (such as through acoustic and visual cues) enables animals to assess predation risk and execute predator-avoidance behavior; however, there are limited studies on the anti-predation behavior of nocturnal animals. The prey of bats provides an excellent representative system for examining the anti-predation behavior of nocturnal animals. Here, we broadcasted different types of echolocation calls of the bird-eating bat Ia io to two wild passerine birds, namely, Zosterops japonicus and Sinosuthora webbiana, that are preyed upon by I. io, and presented the birds with individual bats under different light intensities. The results showed that both bird species were able to perceive the low-frequency audible portion of the bats' echolocation calls; however, they did not exhibit escape responses to the acoustic stimuli. In the dark and under moonlit conditions, both bird species were unable to respond to active bats at close range and the birds only exhibited evasive flight behavior when bats approached or touched them. These results suggest that nocturnal passerine birds may not be able to use acoustic or visual cues to detect bats and adopt evasive maneuvers to avoid predation. This work suggests that bat predation pressure may not elicit primary predator-avoidance responses in nocturnal passerine birds. The results provide new insights into the anti-predation behavior of nocturnal animals.

Linking changes in individual specialization and population niche of space use across seasons in the great evening bat (Ia io).

BACKGROUND: The niche breadth of an animal population comprises both within-individual and between-individual variation (individual specialization). Both components can be used to explain changes in population niche breadth, and this has been extensively investigated in dietary niche dimension studies. However, little is known about how changes in food resources or environmental factors across seasons affect changes in individual and population space use within the same population. METHODS: In this study, we used micro-GPS loggers to capture the space use of individuals and of a population of the great evening bat (Ia io) in summer and autumn. We used I. io as a model to investigate how individual spatial niche breadth and spatial individual specialization affect changes in population niche breadth (home range and core area sizes) across seasons. Additionally, we explored the drivers of individual spatial specialization. RESULTS: We found that the population home range and the core area of I. io did not increase in autumn when insect resources were reduced. Moreover, I. io showed different specialization strategies in the two seasons: higher spatial individual specialization in summer and lower individual specialization but broader individual niche breadth in autumn. This trade-off may maintain the dynamic stability of the population spatial niche breadth across seasons and facilitate the population response to changes in food resources and environmental factors. CONCLUSIONS: Like diet, spatial niche breadth of a population also may be determined by a combination of individual niche breadth and individual specialization. Our work provides new insights into the evolution of niche breadth from the spatial dimension.

Population-level lateralization of boxing displays enhances fighting success in male Great Himalayan leaf-nosed bats.

Behavioral lateralization with left- and right-hand use is common in the Animal Kingdom and can be advantageous for social species. The existence of a preferential use of the hands during agonistic interactions has been described for a number of invertebrate and vertebrate species. Bats compose the second largest order of mammals. They not only use their forelimbs for flight but also agonistic interactions. However, whether bat species show a population-level lateralized aggressive display has largely been unexplored. Here, we examine the lateralization of boxing displays during agonistic interactions in male Great Himalayan leaf-nosed bats, Hipposideros armiger, from three different populations. We found a population-level lateralization of boxing displays: Males from all three populations show a preferential use of the left forearm to attack opponents. In addition, left-handed boxers have higher fighting success over right-handed boxers. This study expands our knowledge of the handedness of bats and highlights the role of behavioral lateralization in conflict resolution in nocturnal mammals.

Seasonal variation of population and individual dietary niche in the avivorous bat, Ia io.

The variation in niche breadth can affect how species respond to environmental and resource changes. However, there is still no clear understanding of how seasonal variability in food resources impacts the variation of individual dietary diversity, thereby affecting the dynamics of a population's dietary niche breadth. Optimal foraging theory (OFT) and the niche variation hypothesis (NVH) predict that when food resources are limited, the population niche breadth will widen or narrow due to increased within-individual dietary diversity and individual specialization or reduced within-individual dietary diversity, respectively. Here, we used DNA metabarcoding to examine the composition and seasonality of diets of the avivorous bat Ia io. Furthermore, we investigated how the dietary niches changed among seasons and how the population niche breadth changed when the availability of insect resources was reduced in autumn. We found that there was differentiation in dietary niches among seasons and a low degree of overlap, and the decrease of insect resource availability and the emergence of ecological opportunities of nocturnal migratory birds might drive dietary niche shifts toward birds in I. io. However, the population's dietary niche breadth did not broaden by increasing the within-individual dietary diversity or individual specialization, but rather became narrower by reducing dietary diversity via predation on bird resources that served as an ecological opportunity when insect resources were scarce in autumn. Our findings were consistent with the predictions of OFT, because birds as prey for bats provided extremely different resources from those of insects in size and nutritional value. Our work highlights the importance of size and quality of prey resources along with other factors (i.e., physiological, behavioral, and life-history traits) in dietary niche variation.

Full-Length Transcriptome of the Great Himalayan Leaf-Nosed Bats (Hipposideros armiger) Optimized Genome Annotation and Revealed the Expression of Novel Genes.

The Great Himalayan Leaf-nosed bat (Hipposideros armiger) is one of the most representative species of all echolocating bats and is an ideal model for studying the echolocation system of bats. An incomplete reference genome and limited availability of full-length cDNAs have hindered the identification of alternatively spliced transcripts, which slowed down related basic studies on bats' echolocation and evolution. In this study, we analyzed five organs from H. armiger for the first time using PacBio single-molecule real-time sequencing (SMRT). There were 120 GB of subreads generated, including 1,472,058 full-length non-chimeric (FLNC) sequences. A total of 34,611 alternative splicing (AS) events and 66,010 Alternative Polyadenylation (APA) sites were detected by transcriptome structural analysis. Moreover, a total of 110,611 isoforms were identified, consisting of 52% new isoforms of known genes and 5% of novel gene loci, as well as 2112 novel genes that have not been annotated before in the current reference genome of H. armiger. Furthermore, several key novel genes, including Pol, RAS, NFKB1, and CAMK4, were identified as being associated with nervous, signal transduction, and immune system processes, which may be involved in regulating the auditory nervous perception and immune system that helps bats to regulate in echolocation. In conclusion, the full-length transcriptome results optimized and replenished existing H. armiger genome annotation in multiple ways and offer advantages for newly discovered or previously unrecognized protein-coding genes and isoforms, which can be used as a reference resource.

Acoustic signal dominance in the multimodal communication of a nocturnal mammal.

Multimodal communication in animals is common, and is particularly well studied in signals that include both visual and auditory components. Multimodal signals that combine acoustic and olfactory components are less well known. Multimodal communication plays a crucial role in agonistic interactions in many mammals, but relatively little is known about this type of communication in nocturnal mammals. Here, we used male Great Himalayan leaf-nosed bats Hipposideros armiger to investigate multimodal signal function in acoustic and olfactory aggressive displays. We monitored the physiological responses (heart rate [HR]) when H. armiger was presented with 1 of 3 stimuli: territorial calls, forehead gland odors, and bimodal signals (calls + odors). Results showed that H. armiger rapidly increased their HR when exposed to any of the 3 stimuli. However, the duration of elevated HR and magnitude of change in HR increased significantly more when acoustic stimuli were presented alone compared with the presentation of olfactory stimuli alone. In contrast, the duration of elevated HR and magnitude of change in HR were significantly higher with bimodal stimuli than with olfactory stimuli alone, but no significant differences were found between the HR response to acoustic and bimodal stimuli. Our previous work showed that acoustic and chemical signals provided different types of information; here we describe experiments investigating the responses to those signals. These results suggest that olfactory and acoustic signals are non-redundant signal components, and that the acoustic component is the dominant modality in male H. armiger, at least as it related to HR. This study provides the first evidence that acoustic signals dominate over olfactory signals during agonistic interactions in a nocturnal mammal.

Feeding behavior innovation increases foraging efficiency in the Amur falcon but may be a threat to Asian particolored bats.

Behavioral innovations are rare and infrequent in the natural world, but they are pivotal for animals to respond to environmental changes. The ecological benefits of these innovations remain unknown, especially in wild populations. Here, two foraging strategies and three eating behaviors of the Amur falcon (Falco amurensis) were observed during predation on Asian particolored bats (Vespertilio sinensis) across 3 years. We demonstrated that an eating behavioral innovation in F. amurensis increased the foraging efficiency of V. sinensis more than twofold during 3 consecutive years. This showed that changes in feeding behavior by a bird strongly influenced the rate of energy intake. Since predation on bats by falcons mainly occurred during the lactation and post-lactation of bats, this may have a certain level of negative effect on the bat population.

Ambush predation and the origin of euprimates.

Primates of modern aspect (euprimates) are characterized by a suite of characteristics (e.g., convergent orbits, grasping hands and feet, reduced claws, and leaping), but the selective pressures responsible for the evolution of these euprimate characteristics have long remained controversial. Here, we used a molecular phyloecological approach to determine the diet of the common ancestor of living primates (CALP), and the results showed that the CALP had increased carnivory. Given the carnivory of the CALP, along with the general observation that orbital convergence is largely restricted to ambush predators, our study suggests that the euprimate characteristics could have been more specifically adapted for ambush predation. In particular, our behavior experiment further shows that nonclaw climbing can significantly reduce noises, which could benefit the ancestral euprimates' stalking to ambush their prey in trees. Therefore, our study suggests that the distinctive euprimate characteristics may have evolved as their specialized adaptation for ambush predation in arboreal environments.

Behavioral innovation and genomic novelty are associated with the exploitation of a challenging dietary opportunity by an avivorous bat.

Foraging on nocturnally migrating birds is one of the most challenging foraging tasks in the animal kingdom. Only three bat species (e.g., Ia io) known to date can prey on migratory birds. However, how these bats have exploited this challenging dietary niche remains unknown. Here, we demonstrate that I. io hunts at the altitude of migrating birds during the bird migration season. The foraging I. io exhibited high flight altitudes (up to 4945 m above sea level) and high flight speeds (up to 143.7 km h-1). I. io in flight can actively prey on birds in the night sky via echolocation cues. Genes associated with DNA damage repair, hypoxia adaptation, biting and mastication, and digestion and metabolism have evolved to adapt to this species' avivorous habits. Our results suggest that the evolution of behavioral innovation and genomic novelty are associated with the exploitation of challenging dietary opportunities.

Comparative analysis of the daily liver transcriptomes in wild nocturnal bats.

BACKGROUND: Mammals rely on the circadian clock network to regulate daily systemic metabolism and physiological activities. The liver is an important peripheral organ in mammals, and it has a unique circadian rhythm regulation process. As the only mammals that can fly, bats have attracted much research attention due to their nocturnal habits and life histories. However, few research reports exist concerning the circadian rhythms of bat liver gene expression and the relevant biological clock regulation mechanisms in the liver. RESULTS: In this study, the expression levels of liver genes of Asian particolored bats were comparatively analyzed using RNA-seq at four different time points across 24 h. A total of 996 genes were found to be rhythmic, accounting for 65% of the total number of expressed genes. The critical circadian rhythm genes Bmal1, Rev-erbα, Cry, and Ror in the liver exhibited different expression patterns throughout the day, and participated in physiological processes with rhythmic changes, including Th17 cell differentiation (ko04659), antigen processing and presentation (ko04612), the estrogen signaling pathway (ko04915), and insulin resistance (ko04931). In addition, previous studies have found that the peroxisome proliferator-activated receptor (PPAR) metabolic signaling pathway (ko03320) may play a vital role in the rhythmic regulation of the metabolic network. CONCLUSIONS: This study is the first to demonstrate diurnal changes in bat liver gene expression and related physiological processes. The results have thus further enriched our understanding of bats' biological clocks.

Extrinsic and intrinsic factors influencing the emergence and return of the Asian particolored bat Vespertilio sinensis to the summer roost.

Circadian rhythms play a crucial role in the health and survival of organisms. However, little is known concerning how intrinsic and extrinsic factors affect animal daily rhythms in the field, especially in nocturnal animals. Here, we investigated the first emergence, mid-emergence, and return times of Vespertilio sinensis, and also integrated environmental conditions (temperature, humidity, and light intensity) and biotic factors (reproductive status and predation risk) to determine causes of variation in the activity rhythms of the bats. We found that variation in the first emergence time, the mid-emergence time, and the final return time were distinct. The results demonstrated that the emergence and return times of bats were affected by light intensity, reproductive status, and predation risk in a relatively complex pattern. Light intensity had the greatest contribution to activity rhythms. Moreover, we first investigated the effects of actual predators on the activity rhythms of bats; the results showed that the mid-emergence time of bats was earlier as predators were hunting, but the final return time was later when predators were present. Finally, our results also highlighted the importance of higher energy demands during the lactation in bats to variation in activity rhythms. These results improve our understanding of the patterns and causes of variation in activity rhythms in bats and other nocturnal animals.

Comparative analysis of the daily brain transcriptomes of Asian particolored bat.

Daily rhythms are found in almost all organisms, and they comprise one of the most basic characteristics of living things. Daily rhythms are generated and mainly regulated by circadian clock. Bats have attracted interest from researchers because of their unique biological characteristics. However, little is known about the molecular underpinnings of daily rhythms in bats. In this study, we used RNA-Seq to uncover the daily rhythms of gene expression in the brains of Asian particolored bats over the 24-h day. Accordingly, four collected time points corresponding to four biological states, rest, sleep, wakefulness, and active, were selected. Several groups of genes with different expression levels in these four states were obtained suggested that different physiological processes were active at various biological states, including drug metabolism, signaling pathways, and the circadian rhythm. Furthermore, downstream analysis of all differentially expressed genes in these four states suggested that groups of genes showed daily rhythms in the bat brain. Especially for Per1, an important circadian clock gene was identified with rhythmic expression in the brain of Asian particolored bat. In summary, our study provides an overview of the brain transcriptomic differences in different physiological states over a 24-h cycle.

The gut microbiota of bats confers tolerance to influenza virus (H1N1) infection in mice.

Pathogens from wild animals cause approximately 60% of emerging infectious diseases (EIDs). Studies on the immune systems of natural hosts are helpful for preventing the spread of EIDs. Bats are natural hosts for many emerging infectious pathogens and have a unique immune system that often coexists with pathogens without infection. Previous studies have shown that some genes and proteins may help bats fight virus infection, but little is known about the function of the bat gut microbiome on immunity. Here, we transplanted gut microbiota from wild bats (Great Himalayan Leaf-nosed bats, Hipposideros armiger) into antibiotic-treated mice, and found that the gut microbiota from bats regulated the immune system faster than mice after antibiotic treatment. Moreover, we infected mice with H1N1, and found that the gut microbiota of bats could effectively protect mice, leading to decreased inflammatory response and increased survival rate. Finally, metabolomics analysis showed that the gut microbiota of bats produced more flavonoid and isoflavones. Our results demonstrate that the quick-start innate immune response endowed by bat gut microbiota and the regulatory and antiviral effects of gut microbiota metabolites successfully ensured mouse survival after viral challenge. To our knowledge, our study was the first to use fecal microbiota transplantation (FMT) to transplant the gut microbiota of bats into mice, and the first to provide evidence that the gut microbiota of bats confers tolerance to viral infections.

Vocal performance reflects individual quality in male Great Himalayan leaf-nosed bats (Hipposideros armiger).

Signals containing parameter trade-offs are likely to be honest indicators of signaler quality because they are difficult to produce. Signals with a trill-rate/bandwidth trade-off have been described for many songbird species, one mouse, and one non-human primate species. However, there were no reports about whether there is a vocal performance trade-off in social calls of bats. This study investigated (1) a possible vocal performance trade-off in territorial calls of male Great Himalayan leaf-nosed bats, Hipposideros armiger, recorded from 9 locations in south China, and (2) the relationships between vocal performance (vocal deviation and consistency) and caller's quality (body mass) to determine whether vocal performance honestly indicates a caller's quality. Vocal deviation measures the deviation of a call relative to an extreme call and vocal consistency measures the spectral consistency across a string of syllables. Our results showed a significant negative correlation between syllable repetition rate and frequency bandwidth, suggesting a vocal performance trade-off similar to the one in songbirds. Further, there was a significant negative relationship between body mass and vocal deviation, but no significant correlation between body mass and vocal consistency. This study provides the first empirical evidence for a vocal performance trade-off of social calls in bats, and the potential for the level of performance to indicate caller quality.

Individuality and function of chemical signals during conflict resolution of a mammal.

Individual recognition via communication signals is a critical component of social behavior, and provides the basis of conflict resolution, territorial behavior, and mate choice. However, the function of chemical signals in mammalian individual recognition and conflict resolution has largely been unexplored despite olfaction being a dominant sensory modality in many mammalian species. Here, we describe behavioral tests designed to evaluate the potential role of forehead gland secretions during conflict related to territorial defense in male Great Himalayan leaf-nosed bats. We used gas chromatography-mass spectrometry to quantify the chemical composition. Our results showed that forehead gland secretions contain 16 categories of compounds, including 84 volatile compounds. The concentrations of compounds and their categories differed significantly among individuals. Moreover, behavioral studies indicated that males can use chemical signals for individual recognition. Contests were staged between males with or without functioning forehead glands. Paired males without functioning forehead glands displayed more physical contact and longer contest duration compared with pairs with functioning glands. Moreover, males with a functioning gland were more likely to win in contests when paired with males without a functioning gland. These findings support a growing amount of evidence that chemical signals play a vital role in conflict resolution in mammals.