Bats are reservoirs for many emerging viruses, yet broad-scale spatial patterns of bat viromes and their ecological determinants remain unclear, limiting spillover risk assessment. To address this, we conducted a standardized survey across central to southern China. During 2022–2023, fecal samples were collected from 527 bats representing 17 species at 21 caves in seven provinces. Using each cave as the analytical unit, samples from all species at a site were pooled to construct 21 fecal RNA virome libraries for metatranscriptomic sequencing; viral-like contigs were annotated, and viral alpha/beta diversity was quantified at the genus level. Sites were grouped by geographic distance and latitude to evaluate spatial differentiation and to relate virome patterns to host community attributes and environmental factors. We annotated 56 viral families, including 19 vertebrate-associated families, with multiple families and genera shared across geographic groupings. Vertebrate-associated viral diversity showed limited evidence of geographic isolation within the study region but suggested gradual turnover with latitude. Host species richness was the strongest correlate of virome diversity, exceeding the effects of evaluated environmental variables (e.g., climate and human activity). These results support prioritization of species-rich bat habitats for surveillance and risk assessment.

Acoustic behavior, essential for communication and perception, is metabolically demanding. Studying the energy costs of echolocation helps us to understand animal energy allocation and provides key insights into the evolutionary constraints of acoustic signals. We examined the constant-frequency bat Rhinolophus nippon using a miniature electrocardiogram system and a custom servomotor that moved prey toward stationary bats. This setup allowed for synchronous recording of high-resolution electrocardiogram and echolocation calls from the search phase to the approach phase. During the search phase, bats emitted isolated echolocation pulses characterized by long pulse durations and inter-pulse intervals (IPIs), together with higher root mean square (RMS) amplitude, pulse energy, and peak amplitude. In the approach phase, call rate increased significantly (3.15-fold), and bats predominantly produced sonar sound groups. Meanwhile, pulse duration, IPIs, RMS amplitude, and pulse energy decreased to 65.23%, 25.82%, 78.50%, and 86.32% of the corresponding search-phase values, whereas peak amplitude increased to 110.99%, indicating that R. nippon can flexibly adjust the structure of its echolocation calls. However, despite the increased call rate (p < 0.05), neither heart rate nor metabolic rate differed between phases. This study provides direct physiological evidence for understanding energy expenditure in bat echolocation and offers a methodological reference for future research.

Bats, as the world’s second-largest mammalian order, have garnered significant attention for their ability to harbor numerous viruses without exhibiting disease symptoms. Nucleotide-binding domain and leucine-rich repeat-containing receptors (NLRs) are crucial components of the immune system. This study conducted an evolutionary analysis of the NLR gene family across 26 bat species to investigate the molecular mechanisms underlying their role in viral resistance under high viral load pressure. We identified gene duplication events in multiple genes. The NLR gene family exhibited high conservation throughout evolution, which may contribute to the occurrence of gene duplication. This conserved genomic structure also ensures functional stability, safeguarding bats’ antiviral resistance. Most NLR genes primarily function within the type I interferon (IFN) signaling pathway and the NF-κB signaling pathway. The NLR gene family enhances the innate immune capacity of bats through the adaptive evolution of some genes, combining enhanced gene functionality with the maintenance of gene conservation at a low evolutionary rate. Moreover, bats employ diverse innate immune strategies, where multiple immune pathways collectively establish the innate immune barrier. The molecular evolution of this gene family provides new insights into the molecular mechanisms and functional pathways involved in the innate immune response of bats.