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.

Identification and genome characterization of novel parechovirus sequences from Hipposideros armiger in China.

BACKGROUND: Bats were identified as a natural reservoir of emerging and re-emerging infectious pathogens threatening human health and life. METHODS: This study collected 21 fecal samples of Hipposideros armiger in Mengla County of Xishuangbanna Prefecture Yunnan Province to combine one pool for viral metagenomic sequencing. RESULTS: Two nearly complete genomes of parechoviruses, BPeV11 and BPeV20, were sequenced. Genome analysis revealed that BPeV11 and BPeV20 follow a 3-3-4 genome layout: 5' UTR-VP0-VP3-VP1-2A-2B-2C-3A-3B-3C-3D-3' UTR. The prevalence of BPev11 and BPev20 by Nested-PCR showed that 1 of 21 fecal samples was positive. Based on amino acid identity comparison and phylogenetic analysis of P1, 2C, and 3D, BPeV11 and BPeV20 were closely related to but distinct from FPeVs. CONCLUSION: It was probably proposed to be a novel species in the genus Parechovirus of the family Picornaviridae. The isolation of BPev11 and BPev20 from H. armiger in China is the first complete genome of parechovirus isolations from bat feces of the genus Hipposideros.

Great Himalayan leaf-nosed bats modify vocalizations to communicate threat escalation during agonistic interactions.

Bats vocalize extensively within various social contexts. Nevertheless, studies of agonistic interactions, associating vocalizations signalling the emotional state of a caller with individual signatures during aggressive vocalizations remain scarce. Here, we examined whether male Great Himalayan leaf-nosed bats (Hipposideros armiger) modify their aggressive vocalizations during agonistic interactions depending on the level of aggression, and whether these vocalizations encode individual signatures. We applied a cost-benefit analysis to audio-video recordings of 50 dyadic agonistic interactions to categorize displays into two levels of aggression intensity (low aggression: bared teeth, slightly pulled up body and/or wings; high aggression: rapidly flapping wings, punching, biting). Male H. armiger used graded visual agonistic displays accompanied by bent upward frequency modulation (bUFM) vocalizations to defend their roosting territories. At high aggression intensities, males decreased the minimum frequency of aggressive calls and increased the frequency bandwidth. Males also transferred energy from the second harmonic to the first harmonic as the threat escalated. These systematic modifications of acoustic parameters as aggressive intensity fluctuated corresponded to prosodic modifications in human speech, indicating that emotion-related acoustic cues are a common feature of acoustic communication in mammals. In addition, we found that the aggressive calls of eight adult males encoded discriminable signatures, and that males could discriminate among individuals based on these aggressive calls. Such discrimination is probably useful for individual identification among rival neighbours.