Seba's Short-tailed Bat Research Articles

Four new species of Anenterotrema Stunkard, 1938 (Digenea: Dicrocoellidae) infecting phyllostomid bats in the Peruvian Amazon.

During a research of digeneans (Platyhelminthes: Trematoda) infecting leaf-nosed bats (Chiroptera: Phyllostomidae) from the Kawsay Biological Station in Madre de Dios, southeastern Peru, four new species were described based on light and scanning electron microscopy (SEM): Anenterotrema paramegacetabulum n. sp. from the seba's short-tailed bat Carollia perspicillata Linnaeus, A. hastati n. sp., A. kawsayense n. sp., and A. peruense n. sp. from the greater spear-nosed bat Phyllostomus hastatus (Pallas). Anenterotrema paramegacetabulum n. sp. differs from all congeners by having a terminal oral sucker, a transversely elongated ventral sucker without clamp-shaped, and by having the testes located immediately posterior to ventral sucker. Anenterotrema hastati n. sp. is easily differentiated from the other congeneric species by having an almost clamp-shaped oral sucker, well-developed cirrus sac, a bilobulated seminal receptacle, and a group of well-developed unicellular glands located anterolateral to cirrus sac. Anenterotrema kawsayense n. sp. is characterized by possessing protuberances on the anterior margin of the oral sucker. Anenterotrema peruense n. sp. is mainly characterized by having the testes located principally anterior to ventral sucker and the cirrus sac located perpendicular to midline of the body. The present finding brings to 12, the number of known species of Anenterotrema. A key to the species of Anenterotrema Stunkard, 1938 is provided.

Demodex carolliae in a colony of Seba's short-tailed bats (Carollia perspicillata): clinical, pathological and parasitological findings

Seba's short-tailed bats (Carollia perspicillata) are a frugivorous species native to Central and South America. Despite their importance as a reservoir for zoonotic pathogens and their popularity in zoological collection and as research models, there are relatively few reports on non-zoonotic diseases of bats. Mites of the genus Demodex are obligate commensals of the skin of a range of mammals, are highly host-specific and are not associated with clinical disease when present in low numbers. However, infestation with high numbers can result in severe or even fatal disease and substantially affect the well-being of the animals. The clinical, pathological and parasitological findings in 12 Seba's short-tailed bats with demodicosis from a colony kept at Munich Zoo Hellabrunn between 1992 and 2021 are described in this report. From 2002, skin lesions became apparent on the head, especially the periocular region, nose and ears, as well as the genital area of some animals. In advanced cases, skin changes were also present on the abdomen, back and extremities. Gross findings typically included alopecia and thickening of the skin, with the formation of papules, reflecting cystically dilated hair follicles containing myriads of demodecid mites. Histologically, lesions were characterized by a paucicellular lymphocytic dermatitis and folliculitis with perifollicular fibrosis, epidermal hyperplasia, orthokeratotic hyperkeratosis and disproportionately high numbers of intrafollicular arthropods. Demodex carolliae was identified morphologically by light, phase-contrast and electron microscopy. Further characterization was achieved by extraction of parasitic DNA and partial gene sequencing of two mitochondrial genes, 16S rDNA and cox1. This is the first clinicopathological description of generalized demodicosis in Seba's short-tailed bats and includes the first molecular characterization of D. carolliae with provision of a GenBank entry.

Flight performance and wing morphology in the bat Carollia perspicillata: biophysical models and energetics.

Studies on functional performance are important to understand the processes responsible for the evolution of diversity. Morphological trait variation within species influences the energetic cost of locomotion and impacts life history traits, with ecological and evolutionary consequences. This study examined wing morphology correlates of flight performance measured by energetic expenditure in the Seba's short-tailed bat, Carollia perspicillata. In the flight experiments, nature caught bats (59 females, 57 males) were allowed to fly for 3 min in a room. After each flight, thermographic images were taken to measure body temperature, and biophysical models were used to calculate sensible heat loss as a measure of energetic expenditure. Wing morphological traits were measured for each individual and associated with heat loss and power required to fly on performance surfaces. Wing morphological traits explained 7-10% of flight energetic cost, and morphologies with the best performance would save the energy equivalent to 9-30% of total daily requirements. The optimal performance areas within the C. perspicillata morphospace were consistent with predicted selection trends from the literature. A trade-off between demands for flight speed and maneuverability was observed. Wing loading and camber presented sexual dimorphism. These morphological differences are likely associated with more economical but less maneuverable flight in females, leading them to fly more often in open areas along the forest edge. Our findings demonstrate how small scale changes in wing morphology can affect life history strategies and fitness.

Oscillatory discharges in the auditory midbrain of the big brown bat contribute to coding of echo delay.

Subsequent to his breakthrough discovery of delay-tuned neurons in the bat's auditory midbrain and cortex, Albert Feng proposed that neural computations for echo delay involve intrinsic oscillatory discharges generated in the inferior colliculus (IC). To explore further the presence of these neural oscillations, we recorded multiple unit activity with a novel annular low impedance electrode from the IC of anesthetized big brown bats and Seba's short-tailed fruit bats. In both species, responses to tones, noise bursts, and FM sweeps contain long latency components, extending up to 60ms post-stimulus onset, organized in periodic, oscillatory-like patterns at frequencies of 360-740Hz. Latencies of this oscillatory activity resemble the wide distributions of single neuron response latencies in the IC. In big brown bats, oscillations lasting up to 30ms after pulse onset emerge in response to single FM pulse-echo pairs, at particular pulse-echo delays. Oscillatory responses to pulses and evoked responses to echoes overlap extensively at short echo delays (5-7ms), creating interference-like patterns. At longer echo delays, responses are separately evident to both pulses and echoes, with less overlap. These results extend Feng's reports of IC oscillations, and point to different processing mechanisms underlying perception of short vs long echo delays.

Cover Image, Volume 32, Issue 10

The cover image is based on the Research Article The retrosplenial cortex of Carollia perspicillata, Seba's short-tailed fruit bat by Timothy Morello et al., https://doi.org/10.1002/hipo.23464.

The retrosplenial cortex of Carollia perspicillata, Seba's short‐tailed fruit bat

Retrosplenial cortex (RSC) is a brain region involved in critical cognitive functions including memory, planning, and spatial navigation and is commonly affected in neurodegenerative diseases. Subregions of RSC are typically described as Brodmann areas 29 and 30, which are defined by cytoarchitectural features. Using immunofluorescence, we studied the distributions of neurons immunoreactive for NeuN, latexin, and calcium binding proteins (calbindin, calretinin, and parvalbumin) in RSC of Carollia perspicillata, Seba's short-tailed fruit bat. We observed that latexin was specifically present in areas 29a and 29b but not 29c and 30. We further identified distribution patterns of calcium binding proteins that group areas 29a and 29b separately from areas 29c and 30. We conclude first that latexin is a useful marker to classify subregions of RSC and second that these subregions contain distinct patterns of neuronal immunoreactivity for calcium binding proteins. Given the long lifespan of Carollia, bat RSC may be a useful model in studying age-related neurodegeneration.

Paramyxoviruses from neotropical bats suggest a novel genus and nephrotropism.

Paramyxoviruses have a broad host range and geographic distribution, including human pathogens transmitted by bats, such as Nipah and Hendra viruses. In this study, we combined high-throughput sequencing and molecular approaches to investigate the presence of paramyxoviruses in neotropical bats (Microchiroptera suborder) in Brazil. We discovered and characterized three novel paramyxoviruses in the kidney tissues of apparently healthy common vampire bats (D. rotundus) and Seba's short-tailed bats (C. perspicillata), which we tentatively named Kanhgág virus (KANV), Boe virus (BOEV), and Guató virus (GUATV). In this study, we classified these viruses as putative species into the Macrojêvirus genus, a newly proposed genus of the Orthoparamyxovirinae subfamily. Using RT-PCR, we detected these viruses in 20.9% (9 out of 43) of bats tested, and viral RNA was detected exclusively in kidney tissues. Attempts to isolate infectious virus were successful for KANV and GUATV. Our results expand the viral diversity, host range, and geographical distribution of the paramyxoviruses.

Malignant Peripheral Nerve Sheath Tumour in a Seba's Short-Tailed Bat (Carollia perspicillata)

There are few reports of tumours in bats, despite the fact that members of the Chiroptera order include 22.4% of all currently known mammal species. We now describe a case of malignant peripheral nerve sheath tumour (MPNST) in an adult female Seba's short-tailed bat (Carollia perspicillata) from a private zoo in Switzerland. The macroscopical and histological findings indicated that the MPNST originated from a dorsal spinal nerve root sheath in the lumbar region and metastasized to the spleen. Immunohistochemical labelling for glial fibrillary acid protein, S100 protein, neuron specific enolase and synaptophysin was negative. The cytoplasm of a few neoplastic cells labelled for smooth muscle actin and desmin. To our knowledge, this is the first report of a PNST in a member of the Chiroptera order.

Functional skull asymmetries in Carollia perspicillata (Phyllostomidae Gray, 1825: Carollinae)

Analysing asymmetry in wild or domestic species enables the evaluation of the morphological responses to functional requirements and/or stress. This report is a study of the cranial asymmetry of Seba's short-tailed bat Carollia perspicillata by means of geometric morphometric techniques. We studied three types of bilateral asymmetries -fluctuating asymmetry, directional asymmetry and antisymmetry- using 15 skull landmarks on dorsal aspect of 45 skulls of Carollia perspicillata (21 males and 24 females) from different localities in Colombia. Directional asymmetry appeared to be significant and clearly higher than fluctuating asymmetry, with the braincase presenting the larger variance. There were no differences between sexes. Echolocation has a great anatomical effect on the bat cranium, and this would explain detected asymmetry.

Hepatozoon Infecting Bats in the Southeastern Brazilian Rainforest.

Tick-borne protozoans of the genus Hepatozoon are obligate hemoparasites that can infect domestic and wild terrestrial vertebrates. Main hepatozoonosis affects canids and involves mainly Hepatozoon canis and Hepatozoon americanum. However, molecular studies revealed the capacity of H. canis to infect a wide range of wild mammals. In July 2018, we conducted an epidemiological survey for tick-borne pathogens in wild hosts, assaying Hepatozoon sp. occurrence in 34 bats captured in different habitats within a conservation unit in the state of Espírito Santo, southeastern Brazil. Blood and spleen tissue DNA samples were submitted to PCR amplifications of Babesia/Theileria and Hepatozoon 18S rRNA gene and 21% (7/34) were positive for Hepatozoon sp. Phylogenetic inferences grouped the obtained sequences from Seba's short-tailed bat (Carollia perspicillata) with the H. canis cluster, and from the great fruit-eating bat (Artibeus lituratus) with rodent-associated Hepatozoon cluster. Further studies are needed to characterize the epidemiological role of Seba's short-tailed bat and the great fruit-eating bat in the wild transmission cycle of these hemoparasites in Brazil.

High Specificity and Aggregation, But Low Prevalence in Bat-Fly Interactions in an Environmental Protection Area in Brazil

Nycteribiids and streblids are obligate hematophagous flies restricted to bats. These flies are widely distributed throughout the Neotropical region, although little is known about their spatial distribution and patterns in parasitic associations in different environments. Studies assessing bat-fly interactions are mostly qualitative, but quantitative studies that allow testing and/or establishing more specific patterns for these interactions are still scarce. Here, we qualitatively and quantitatively assessed the bat-fly interactions in an environmental protection area in northeastern Brazil using prevalence, mean intensity of infestation, aggregation, and abundance indices. We captured 654 bats of 21 species and five families. Only 157 bats (24%) were parasitized by 351 flies of 19 species and eight genera. Among the most representative bat species (n ≥ 15), the host-parasite relationship between the white-lined broad-nosed bat (Platyrrhinus lineatus) and a streblid fly Trichobius angulatus had the highest prevalence (33.3%). The highest mean intensity of infestation recorded was for Trichobius joblingi on the Seba's short-tailed bat (Carollia perspicillata) (2.05 flies per host). The fly with higher values of aggregation was Strebla guajiro (D = 0.98) on C. perspicillata. Ninety-five percent of the fly species were highly host-specific (monoxenous). Regarding the mean abundance of flies, we found no differences between dry and rainy periods. No noticeable pattern for northeastern Brazil was observed when comparing our results with previous studies, except for specificity that was consistently high throughout the region. Our study raises questions about the pattern of fly parasitism on bats and may guide future research on this subject.

Recovering from a tumble and roosting bat style

Evolution has had some fun with the limbs that we call arms. From weight-bearing forelimbs, to fins and flippers, the same skeletal structures keep turning up; albeit somewhat modified. When David Boerma from Brown University, USA, gazes at the wings of a bat, he says that he literally sees himself; ‘from an anatomical perspective their wings aren't very different from my own arms and hands’, he smiles. Having traded in their manual dexterity, bats mastered an airborne lifestyle, but Boerma wondered how the mammal's elegantly elongated limbs might contribute as they navigate natural gusts and squalls. Knowing that bat wings are relatively heavy, Boerma wondered whether the mammals could capitalize on the limbs’ apparent weighty inertia to stabilise their manoeuvres when knocked off course by a gust of wind.Working with Tim Treskatis, an intern from Westphalian University of Applied Sciences, Germany, to build a laser-triggered blast of air that would unbalance Seba's short-tailed bats (Carollia perspicillata) as they flew along a narrow corridor in the lab, Boerma filmed the bats’ recoveries in 3D to reconstruct their manoeuvres. Remarkably, when Boerma, Treskatis, Kenneth Breuer and Sharon Swartz analysed the bats’ movements they were amazed that the animals were able to recover from being thrust off course within a single wing beat.Focusing on the bats that had been pitched upward when the 2.5× bodyweight puff of air hit their back end, the bats flung their wings wide at a point in the wing beat when they would normally pull them in; with the result that their wings were thrust forward, to counteract the jet of air and tip the body back down. However, when the jet glanced off one of the bats’ wings – forcing it down and sending the animal into a spin – the bats swiftly extended the unperturbed wing as it rolled upward in a bid to counteract the rotation. Then they swept the perturbed wing all the way under the body as the bat righted itself ready to resume normal wing beats. ‘By just shifting the weight of their wings around, bats can generate forces that rapidly reorient their bodies when recovering from aerial stumbles, similar to the way a gymnast or diver moves their arms and legs to spin in mid-air’, says Boerma.By this point, Boerma's appetite for research had been whetted; ‘the experience left me yearning to work with bats in the field, where I might see first-hand the kinds of environmental challenges they face and learn how they respond’, he recalls. Explaining that Spix's disk-winged bats (Thyroptera tricolor), which roost head-up inside furled Heliconia leaves in the rainforest, have evolved unconventional suction cups at the base of the thumb and at the ankle for attachment to smooth leaves, Boerma was curious how the minute mammals manage their inbound trajectory when coming in to land.Travelling to the Hacienda Baru Biological Research Station, Costa Rica, and teaming up with José Pablo Barrantes from the Universidad de Costa Rica, the duo spent two weeks tracking down the tiny mammals in their rainforest home. Returning with them briefly to the research station, the pair quickly trained the bats to land on a force plate disguised as a leaf while filming the manoeuvre. And Boerma recalls how the bats were equally happy to land on the researchers as they were on the force plate: ‘At one point José Pablo left the enclosure and I kept training the bat on my own. It flew around and then landed on my back where I couldn't reach it’, he laughs. Analysing movies of the bats landing on the force plate and within furled leaves, with the help of high-school student Charles Chung, Boerma realised that the bats fine-tune their approach by subtly adjusting their wing beats until just above the landing site. Then they pull in their wings, going into freefall as they reach forward with their thumb suction pads to latch onto the surface. Evaluating the force exerted on the force plate as the bat made contact, the duo also realised that the suction pad snaps into place as soon as it contacts the leaf.‘These kinds of studies help us to begin to apply what we learn about bat flight in the lab to understand the more complex flight they perform in their natural habitats’, says Boerma. He is now eager to understand how reeling bats coordinate their deft manoeuvres; in addition to looking for connections between the landing styles of other species to learn how the mammals have adapted to roost sites ranging from tightly furled leaves to caves and trees.

Can extreme MHC class I diversity be a feature of a wide geographic range? The example of Seba's short-tailed bat (Carollia perspicillata).

The major histocompatibility complex (MHC) is one of the most diverse genetic regions under pathogen-driven selection because of its central role in antigen binding and immunity. The highest MHC variability, both in terms of the number of individual alleles and gene copies, has so far been found in passerine birds; this is probably attributable to passerine adaptation to both a wide geographic range and a diverse array of habitats. If extraordinary high MHC variation and duplication rates are adaptive features under selection during the evolution of ecologically and taxonomically diverse species, then similarly diverse MHC architectures should be found in bats. Bats are an extremely species-rich mammalian group that is globally widely distributed. Many bat species roost in multitudinous groups and have high contact rates with pathogens, conspecifics, and allospecifics. We have characterized the MHC class I diversity in 116 Panamanian Seba’s short-tailed bats (Carollia perspicillata), a widely distributed, generalist, neotropical species. We have detected a remarkable individual and population-level diversity of MHC class I genes, with between seven and 22 alleles and a unique genotype in each individual. This diversity is comparable with that reported in passerine birds and, in both taxonomic groups, further variability has evolved through length polymorphisms. Our findings support the hypothesis that, for species with a geographically broader range, high MHC class I variability is particularly adaptive. Investigation of the details of the underlying adaptive processes and the role of the high MHC diversity in pathogen resistance are important next steps for a better understanding of the role of bats in viral evolution and as carriers of several deadly zoonotic viruses.Electronic supplementary materialThe online version of this article (10.1007/s00251-019-01128-7) contains supplementary material, which is available to authorized users.

Food restriction, but not seasonality, modulates the acute phase response of a Neotropical bat

Season and food intake are known to affect immune response of vertebrates yet their effects on metabolic rate have been rarely explored. We tested the effect of season and acute food restriction and their interaction on the energetic cost of immune response activation of a tropical vertebrate, the Seba's short-tailed fruit bat (Carollia perspicillata). We specifically stimulated the acute phase response (APR) with bacterial lipopolysaccharide (LPS) to measure metabolic changes along with changes in body temperature (Tb), body mass (Mb), white blood cell counts and the Neutrophil/Lymphocyte ratio (N/L). We found no effect of season on the different factors associated to the activation of the APR. In contrast to our expectations, unfed bats reached similar Tb increments and RMR peak values and had higher RMR scope values and higher caloric costs than fed bats after LPS injection. However, food deprivation led to delayed metabolic response indicated by longer time required to reach peak RMR values in unfed bats. Both food-deprived and fed bats did not present leukocytosis after APR activation and their WBC counts were similar, but unfed bats had a significant increase of N/L. APR activation represented a small fraction of the bat daily energy requirements which might explain why unfed bats were not limited to mount a metabolic response. Our study adds to recent evidence showing that activating the innate immune system is not an energetically expensive process for plant-eating bats.

Dietary variation during reproduction in Seba’s short-tailed fruit bat

Energy and nutrient availability are important determinants of fetal and offspring development. Given high energy and nutrient expenditure during pregnancy and lactation, foraging strategies and choices of specific food items are critical for survival of both mother and offspring. We analyzed dietary variation and consumption of nitrogen-rich and calcium-rich foods during different developmental stages in Seba's short-tailed fruit bat (Carollia perspicillata). Dietary analysis was carried out on fecal samples from males and nonpregnant, early pregnant, late pregnant, and lactating females at Macaregua Cave in Santander, Colombia. Diets were similar among males, nonpregnant females, and pregnant females, but lactating females exhibited statistically distinguishable diets. Males and pregnant females displayed a generalist diet, eating a wide variety of items, whereas nonpregnant and lactating females had narrower diets. Nitrogen-rich items, such as Piper (Piperales: Piperaceae; pepper plants) and insects, were not prevalent in the diet of pregnant bats, whereas spiked pepper exceeded expected frequencies in the diets of lactating bats. Calcium-rich Ficus (Rosales: Moraceae; figs) was rarely found in the diets of lactating or pregnant bats. Increased calcium requirements associated with lactation may therefore be satisfied through other avenues.

Modification of sperm quality after sexual abstinence in Seba's short-tailed bat, Carollia perspicillata.

In polygynous mating systems, few males have stable access to sexual mates. With an expected higher copulation rate, harem males may deplete seminal fluids or increase epididymal sperm maturation, generating poor sperm quality. In a first study, we reported a higher sperm quality in sneaker males of Carollia perspicillata To test whether the lower sperm quality observed in harem males was generated by an elevated copulation rate, we temporarily removed males of both social statuses from the colony. We thus assessed status-related changes of sperm quality resulting from sexual abstinence. Moreover, released from territory and female guarding, harem males were expected to show a reduction in somatic costs. On the basis of sperm competition models, we predicted a higher resource investment in the ejaculate with the reduction of pre-copulatory efforts. In line with our predictions, sperm quality of harem males improved significantly in contrast to sneaker males, whose sperm quality did not change. Without an increase in ejaculate lipid peroxidation, our results also provide evidence that the duration of sexual abstinence was not sufficient to generate sperm oxidative damage through senescence. Harem males did not show a reduction in blood lipid peroxidation or in the ratio of oxidized to reduced glutathione. In line with the maintenance of these somatic costs, harem males did not invest more superoxide dismutase to the ejaculate to maintain sperm quality. Our results suggest that a difference in copulation rate rather than an adaptation to sperm competition provides sneaker males with higher sperm quality in C. perspicillata.

Sneaker male athletic sperm outstrips harem male sperm to get egg

![Figure][1] Seba's short-tailed bat. Photo credit: Emmanuel Rey. When Nicolas Fasel, from the University of Bern, Switzerland, talks about bat reproduction, it sounds more like war than a love story. Having established their supremacy over a territory, dominant ‘harem’ males then have

When bats are boxing: aggressive behaviour and communication in male Seba's short-tailed fruit bat

Aggressive behaviours have an important impact on the social organization of animals and on the social status of individuals, especially in gregarious species. Agonistic interactions between territory holders are essential to set and reinforce territorial borders. Additionally, agonistic displays are used to demonstrate ownership of a territorial site and may indicate social status of the signaller. Between neighbouring territory owners, dynamic borders require frequent interactions. In daily repeated aggressive encounters, ritualization can help to avoid the need for serious fights and their costly consequences. In the bat Carollia perspicillata, a gregarious frugivore with resource defence polygyny, males defend territories at valuable roosting sites for females. Working with a captive bat colony of about 400 individuals, housed under seminatural conditions in a tropical zoo, we found that males defended territories aggressively by the use of a succession of displays forming a ritualized structure. Simultaneously, males used three different vocalization types during aggressive displays, namely down-sweeps, warbles and aggressive trills. A statistical analysis of 58 aggressive trills from five adult males showed that they contained sufficient variation to encode an individual signature. Using a habituation–dishabituation paradigm playback experiment, we found that males could discriminate between vocalizing males based on aggressive calls alone. Such discrimination is probably useful for distinguishing between neighbouring territory owners and more unfamiliar intruders, and thus allows for an economical response.

Bat airway epithelial cells: a novel tool for the study of zoonotic viruses.

Bats have been increasingly recognized as reservoir of important zoonotic viruses. However, until now many attempts to isolate bat-borne viruses in cell culture have been unsuccessful. Further, experimental studies on reservoir host species have been limited by the difficulty of rearing these species. The epithelium of the respiratory tract plays a central role during airborne transmission, as it is the first tissue encountered by viral particles. Although several cell lines from bats were established recently, no well-characterized, selectively cultured airway epithelial cells were available so far. Here, primary cells and immortalized cell lines from bats of the two important suborders Yangochiroptera and Yinpterochiroptera, Carollia perspicillata (Seba's short-tailed bat) and Eidolon helvum (Straw-colored fruit bat), were successfully cultured under standardized conditions from both fresh and frozen organ specimens by cell outgrowth of organ explants and by the use of serum-free primary cell culture medium. Cells were immortalized to generate permanent cell lines. Cells were characterized for their epithelial properties such as expression of cytokeratin and tight junctions proteins and permissiveness for viral infection with Rift-Valley fever virus and vesicular stomatitis virus Indiana. These cells can serve as suitable models for the study of bat-borne viruses and complement cell culture models for virus infection in human airway epithelial cells.

Mother–offspring recognition in the bat Carollia perspicillata

Parental care is crucial for offspring survival in many taxa but its burden and costs are often not equally distributed between the sexes. In bats, the majority of parental care is provided by females, making examples of paternal support towards pups exceedingly rare. One exception to this general pattern seemed to be the polygynous Seba's short-tailed fruit bat Carollia perspicillata; two earlier studies suggested that paternal care occurs, i.e. that harem males prompt females to retrieve vocalizing pups. To corroborate this suggestion, we investigated the occurrence of maternal and paternal care in reaction to pup isolation calls in C. perspicillata. Acoustic measurements of 905 isolation calls from 17 pups revealed sufficient interindividual variation to encode an individual vocal signature. Correspondingly, mothers were capable of using this individual signature to discriminate between their own pups and age-matched pups from other females belonging to their colony. Maternal experience was positively correlated with the strength of response behaviour during playbacks. Thus, our results indicate that pup isolation calls were used to elicit maternal care and that mothers recognize their pups based on an individual signature in isolation calls. However, in contrast to the previous studies mentioned above, we found no evidence that harem males reacted to pup isolation calls by prompting the respective mothers to retrieve their vocalizing pups. Instead, our results demonstrate that harem males engaged in courtship activities that were unaffected by pup isolation calls.