Vocal correlates of arousal in bottlenose dolphins (Tursiops spp.) in human care.

Human-controlled regimes can entrain behavioural responses and may impact animal welfare. Therefore, understanding the influence of schedules on animal behaviour can be a valuable tool to improve welfare, however information on behaviour overnight and in the absence of husbandry staff remains rare. Bottlenose dolphins (Tursiops spp.) are highly social marine mammals and the most common cetacean found in captivity. They communicate using frequency modulated signature whistles, a whistle type that is individually distinctive and used as a contact call. We investigated the vocalisations of ten dolphins housed in three social groups at uShaka Sea World dolphinarium to determine how patterns in acoustic behaviour link to dolphinarium routines. Investigation focused on overnight behaviour, housing decisions, weekly patterns, and transitional periods between the presence and absence of husbandry staff. Recordings were made from 17h00 - 07h00 over 24 nights, spanning May to August 2018. Whistle (including signature whistle) presence and production rate decreased soon after husbandry staff left the facility, was low over night, and increased upon staff arrival. Results indicated elevated arousal states particularly associated with the morning feeding regime. Housing in the pool configuration that allowed observation of staff activities from all social groups was characterised by an increase in whistle presence and rates. Heightened arousal associated with staff presence was reflected in the structural characteristics of signature whistles, particularly maximum frequency, frequency range and number of whistle loops. We identified individual differences in both production rate and the structural modification of signature whistles under different contexts. Overall, these results revealed a link between scheduled activity and associated behavioural responses, which can be used as a baseline for future welfare monitoring where changes from normal behaviour may reflect shifts in welfare state.

It's not all about the Soprano: Rhinolophid bats use multiple acoustic components in echolocation pulses to discriminate between conspecifics and heterospecifics.

Acoustic communication plays a pivotal role in conspecific recognition in numerous animal taxa. Vocalizations must therefore have discrete acoustic signatures to facilitate intra-specific communication and to avoid misidentification. Here we investigate the potential role of echolocation in communication in horseshoe bats. Although it has been demonstrated that echolocation can be used to discriminate among con- and hetero-specifics, the specific acoustic cues used in discrimination are still relatively unknown. Furthermore, the Acoustic Communication Hypothesis proposes that in multispecies assemblages, in which echolocation frequencies are likely to overlap, bats partition acoustic space along several dimensions so that each species occupies a discrete communication domain. Thus, multiple echolocation variables may be used in discrimination. The objective of this study was to investigate the potential of various echolocation variables to function as discriminatory cues in echolocation-based species discrimination. Using habituation-dishabituation playback experiments, we firstly tested the ability of Rhinolophus clivosus to discriminate between echolocation pulses of heterospecifics with either discrete or overlapping frequencies. Secondly, to determine whether R. clivosus could use echolocation variables other than frequency, we investigated its ability to discriminate among echolocation pulses differing in only one manipulated parameter. These test variables were identified by their contribution to the dissimilarity among pulses. Our results suggest that R. clivosus could discriminate readily between species using echolocation pulses with discrete frequencies. When frequencies overlapped, the ability of bats to discriminate was dependant on additional acoustic variables that defined the acoustic space occupied by the test signal. These additional acoustic variables included, but may not be restricted to, sweep rate of the FM and duty cycle. Thus, when echolocation pulses share a similar acoustic domain, bats use several cues to reliably discriminate among heterospecifics.

Nuclear introns outperform mitochondrial DNA in inter-specific phylogenetic reconstruction: Lessons from horseshoe bats (Rhinolophidae: Chiroptera).

Despite many studies illustrating the perils of utilising mitochondrial DNA in phylogenetic studies, it remains one of the most widely used genetic markers for this purpose. Over the last decade, nuclear introns have been proposed as alternative markers for phylogenetic reconstruction. However, the resolution capabilities of mtDNA and nuclear introns have rarely been quantified and compared. In the current study we generated a novel ∼5kb dataset comprising six nuclear introns and a mtDNA fragment. We assessed the relative resolution capabilities of the six intronic fragments with respect to each other, when used in various combinations together, and when compared to the traditionally used mtDNA. We focused on a major clade in the horseshoe bat family (Afro-Palaearctic clade; Rhinolophidae) as our case study. This old, widely distributed and speciose group contains a high level of conserved morphology. This morphological stasis renders the reconstruction of the phylogeny of this group with traditional morphological characters complex. We sampled multiple individuals per species to represent their geographic distributions as best as possible (122 individuals, 24 species, 68 localities). We reconstructed the species phylogeny using several complementary methods (partitioned Maximum Likelihood and Bayesian and Bayesian multispecies-coalescent) and made inferences based on consensus across these methods. We computed pairwise comparisons based on Robinson-Foulds tree distance metric between all Bayesian topologies generated (27,000) for every gene(s) and visualised the tree space using multidimensional scaling (MDS) plots. Using our supported species phylogeny we estimated the ancestral state of key traits of interest within this group, e.g. echolocation peak frequency which has been implicated in speciation. Our results revealed many potential cryptic species within this group, even in taxa where this was not suspected a priori and also found evidence for mtDNA introgression. We demonstrated that by using just two introns one can recover a better supported species tree than when using the mtDNA alone, despite the shorter overall length of the combined introns. Additionally, when combining any single intron with mtDNA, we showed that the result is highly similar to the mtDNA gene tree and far from the true species tree and therefore this approach should be avoided. We caution against the indiscriminate use of mtDNA in phylogenetic studies and advocate for pilot studies to select nuclear introns. The selection of marker type and number is a crucial step that is best based on critical examination of preliminary or previously published data. Based on our findings and previous publications, we recommend the following markers to recover phylogenetic relationships between recently diverged taxa (<20 My) in bats and other mammals: ACOX2, COPS7A, BGN, ROGDI and STAT5A.

Phenotypic convergence in genetically distinct lineages of a Rhinolophus species complex (Mammalia, Chiroptera).

Phenotypes of distantly related species may converge through adaptation to similar habitats and/or because they share biological constraints that limit the phenotypic variants produced. A common theme in bats is the sympatric occurrence of cryptic species that are convergent in morphology but divergent in echolocation frequency, suggesting that echolocation may facilitate niche partitioning, reducing competition. If so, allopatric populations freed from competition, could converge in both morphology and echolocation provided they occupy similar niches or share biological constraints. We investigated the evolutionary history of a widely distributed African horseshoe bat, Rhinolophus darlingi, in the context of phenotypic convergence. We used phylogenetic inference to identify and date lineage divergence together with phenotypic comparisons and ecological niche modelling to identify morphological and geographical correlates of those lineages. Our results indicate that R. darlingi is paraphyletic, the eastern and western parts of its distribution forming two distinct non-sister lineages that diverged ~9.7 Mya. We retain R. darlingi for the eastern lineage and argue that the western lineage, currently the sub-species R. d. damarensis, should be elevated to full species status. R. damarensis comprises two lineages that diverged ~5 Mya. Our findings concur with patterns of divergence of other co-distributed taxa which are associated with increased regional aridification between 7-5 Mya suggesting possible vicariant evolution. The morphology and echolocation calls of R. darlingi and R. damarensis are convergent despite occupying different biomes. This suggests that adaptation to similar habitats is not responsible for the convergence. Furthermore, R. darlingi forms part of a clade comprising species that are bigger and echolocate at lower frequencies than R. darlingi, suggesting that biological constraints are unlikely to have influenced the convergence. Instead, the striking similarity in morphology and sensory biology are probably the result of neutral evolutionary processes, resulting in the independent evolution of similar phenotypes.

Affect cues in vocalizations of the bat, Megaderma lyra, during agonistic interactions.

Some features of emotional prosody in human speech may be traced back to affect cues in mammalian vocalizations. The present study addresses the question whether affect intensity, as expressed by the intensity of behavioral displays, is encoded in vocal cues, i.e., changes in the structure of associated calls, in bats, a group evolutionarily remote from primates. A frame-by-frame video analysis of 109 dyadic agonistic interactions recorded in approach situations was performed to categorize displays into two intensity levels based on a cost-benefit estimate. M. lyra showed graded visual displays accompanied by specific calls and response calls of the second bat. A sound analysis revealed systematic changes of call sequence parameters with display level. At the high intensity level, total call duration, number of syllables within a call, and the number of calls within a sequence were increased, while intervals between call syllables were decreased for both call types. In addition, the latency of the response call was shorter, and its main syllable-type durations and fundamental frequency were increased. These systematic changes of vocal parameters with affect intensity correspond to prosodic changes in human speech, suggesting that emotion-related acoustic cues are a common feature of vocal communication in mammals.