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Assessing the state of aboveground ecosystems is often possible using remotely sensed data and subterranean systems are often left in the dark. Caves and subterranean habitats represent distinct ecosystems which harbour unique species but are sensitive to different pressures. Given the challenge of assessing many taxa, bats are ideal ecological indicators of caves and subterranean ecosystems, because they serve as keystone species that facilitate energy flow to other life forms inside caves. We calibrated the Bat Cave Vulnerability Index 3.0 (BCVI-S) and its indicators to optimise the scalability and applicability of the index in wider contexts by improving the calculations of Biotic Potential and the use of more standardised remotely sensed data to obtain Biotic Vulnerability based on landscape features. We demonstrated the application of the BCVI-S and the differences in cave priority levels in terms of Biotic Potential, Biotic Vulnerability, and overall priority level. The BCVI-S was more sensitive to the presence of rare and threatened species while balancing the priority level of caves with a hyperabundant population of common species. Simultaneously, the BCVI-S showed higher congruence with the commonly used biodiversity indices than the BCVI-L. More importantly, our calibrated BCVI-S allows users and assessors to easily integrate priorities at different spatiotemporal scales and consider other contexts of bat caves. The updated index is user-friendly and offers a comprehensive standard method for identifying key caves for conservation, which can be integrated into broader national objectives and strategies.

Rodents represent over 40% of known mammal species and are found in various terrestrial habitats. They are significant reservoirs for zoonotic viruses, including harmful pathogens such as arenaviruses and hantaviruses, yet knowledge of their hosts and distributions is limited. Therefore, characterizing the virome profile in these animals is invaluable for outbreak preparedness, especially in potential hotspots of mammal diversity. This study included 681 organs from 124 rodents and one Chinese tree shrew collected from Yunnan Province, China, during 2020-2021. Metagenomic analysis revealed unique features of mammalian viruses in rodent organs across habitats with varying human disturbances. R. tanezumi in locations with high anthropogenic disturbance exhibited the highest mammal viral diversity, with spleen and lung samples showing the highest diversities for these viruses at the organ level. Mammal viral diversity for both commensal and non-commensal rats was identified to positively correlate with landscape disturbance. Some virus families were associated with particular organs or host species, suggesting tropism for these pathogens. Notably, known and novel viral species that are likely to infect humans were identified. R. tanezumi was identified as a reservoir and carrier for various zoonotic viruses, including porcine bocavirus, hantavirus, cardiovirus, and lyssavirus. These findings highlight the influence of rodent community composition and anthropogenic activities on diverse virome profiles, with R. tanezumi as an important reservoir for zoonotic viruses.

Ectoparasites found on bats are known to contain important microbes. However, the viruses hosted by these obligate parasites are understudied. This has led to the near oversight of the potential role of these ectoparasites in virus maintenance and transmission from bats to other interacting species and the environment. Here, we sampled bat ectoparasites parasitizing a diverse selection of bat species in the families Rhinolophidae, Vespertilionidae, Megadermatidae, Hipposideridae and Pteropodidae in Yunnan Province, China. We show that the ectoparasite prevalence was generally higher in male compared to female bats. Most ectoparasites were found to fall within the Nycteribiidae, Spinturnicidae and Streblidae bat ectoparasite families. We subsequently applied a non-biased sequencing of libraries prepared from the pooled ectoparasites, followed by an in-silico virus-centric analysis of the resultant reads. We show that ectoparasites hosted by the sampled families of bats are found to carry, in addition to a diverse set of phages, vertebrate and insect viruses in the families Aliusviridae, Ascoviridae, Chuviridae, Circoviridae, Flaviviridae, Hepadnaviridae, Hepeviridae, Herpesviridae, Iridoviridae, Marseilleviridae, Nairoviridae, Orthomyxoviridae, Parvoviridae, Poxviridae, Reoviridae, Retroviridae, and Rhabdoviridae. We further report a partial Parvovirus VP1/VP2 gene and partial Poxvirus ubiquitin-like gene predicted by two independent next generation sequencing data analysis pipelines. This study describes the natural virome of bat ectoparasites, providing a platform for understanding the role these ectoparasites play in the maintenance and spread of viruses to other animals.

Abstract Background Family Rhinolophidae (horseshoe bats), Hipposideridae (leaf-nosed bats) and Rhinonycteridae (trident bats) are exclusively distributed in the Old-World, and their biogeography reflects the complex historic geological events throughout the Cenozoic. Here we investigated the origin of these families and unravel the conflicting family origin theories using a high resolution tree covering taxa from each zoogeographic realm from Africa to Australia. Ancestral range estimations were performed using a probabilistic approach implemented in BioGeoBEARS with subset analysis per biogeographic range [Old-World as whole, Australia–Oriental–Oceania (AOO) and Afrotropical–Madagascar–Palearctic (AMP)]. Result Our result supports an Oriental origin for Rhinolophidae, whereas Hipposideridae originated from the Oriental and African regions in concordance with fossil evidence of both families. The fossil evidence indicates that Hipposideridae has diversified across Eurasia and the Afro-Arabian region since the Middle Eocene. Meanwhile, Rhinonycteridae (the sister family of Hipposideridae) appears to have originated from the Africa region splitting from the common ancestor with Hipposideridae in Africa. Indomalaya is the center of origin of Rhinolophidae AOO lineages, and Indomalayan + Philippines appears to be center of origin of Hipposideridae AOO lineage indicating allopatric speciation and may have involved jump-dispersal (founder-event) speciation within AOO lineage. Wallacea and the Philippines may have been used as stepping stones for dispersal towards Oceania and Australia from the Oriental region. Multiple colonization events via different routes may have occurred in the Philippines (i.e., Palawan and Wallacea) since the Late Miocene. The colonization of Rhinolophidae towards Africa from Asia coincided with the estimated time of Tethys Ocean closure around the Oligocene to Miocene (around 27 Ma), allowing species to disperse via the Arabian Peninsula. Additionally, the number of potential cryptic species in Rhinolophidae in Southeast Asia may have increased since Plio-Pleistocene and late Miocene. Conclusion Overall, we conclude an Oriental origin for Rhinolophidae, and Oriental + African for Hipposideridae. The result demonstrates that complex historical events, in addition to species specific ecomorphology and specialization of ecological niches may shape current distributions.

Measurement(s) spatial region Technology Type(s) occurrent Sample Characteristic - Organism Mammalia • Chiroptera sp. BOLD:AAA2524 Sample Characteristic - Environment cave system • karst cave Sample Characteristic - Location Global

Abstract Background In genus Rhinolophus, species in the Rhinolophus philippinensis and R. macrotis groups are unique because the horseshoe bats in these group have relatively low echolocation frequencies and flight speeds compared with other horseshoe bats with similar body size. The different characteristics among bat species suggest particular evolutionary processes may have occurred in this genus. To study the adaptive evidence in the mitochondrial genomes (mitogenomes) of rhinolophids, especially the mitogenomes of the species with low echolocation frequencies, we sequenced eight mitogenomes and used them for comparative studies of molecular phylogeny and adaptive evolution. Results Phylogenetic analysis using whole mitogenome sequences produced robust results and provided phylogenetic signals that were better than those obtained using single genes. The results supported the recent establishment of the separate macrotis group. The signals of adaptive evolution discovered in the Rhinolophus species were tested for some of the codons in two genes (ND2 and ND6) that encode NADH dehydrogenases in oxidative phosphorylation system complex I. These genes have a background of widespread purifying selection. Signals of relaxed purifying selection and positive selection were found in ND2 and ND6, respectively, based on codon models and physicochemical profiles of amino acid replacements. However, no pronounced overlap was found for non-synonymous sites in the mitogenomes of all the species with low echolocation frequencies. A signal of positive selection for ND5 was found in the branch-site model when R. philippinensis was set as the foreground branch. Conclusions The mitogenomes provided robust phylogenetic signals that were much more informative than the signals obtained using single mitochondrial genes. Two mitochondrial genes that encoding proteins in the oxidative phosphorylation system showed some evidence of adaptive evolution in genus Rhinolophus and the positive selection signals were tested for ND5 in R. philippinensis. These results indicate that mitochondrial protein-coding genes were targets of adaptive evolution during the evolution of Rhinolophus species, which might have contributed to a diverse range of acoustic adaptations in this genus.

Systematics and taxonomy are the backbone of all components of biology and ecology, yet cryptic species present a major challenge for accurate species identification. This is especially problematic as they represent a substantial portion of undiscovered biodiversity, and have implications for not only species conservation, but even assaying potential risk of zoonotic spillover. Here, we use integrative approaches to delineate potential cryptic species in horseshoe bats (Rhinolophidae), evaluate the phenotypic disparities between cryptic species, and identify key traits for their identification. We tested the use of multispecies coalescent models (MSC) using Bayesian Phylogenetic and Phylogeography (BPP) and found that BPP was useful in delineating potential cryptic species, and consistent with acoustic traits. Our results show that around 40% of Asian rhinolophid species are potentially cryptic and have not been formally described. In order to avoid potential misidentification and allow species to be accurately identified, we identified quantitative noseleaf sella and acoustic characters as the most informative traits in delineating between potential cryptic species in Rhinolophidae. This highlights the physical differences between cryptic species that are apparent in noseleaf traits which often only qualitatively described but rarely measured. Each part of the noseleaf including the sella, lateral lappets, and lancet furrows, play roles in focusing acoustic beams and thus, provide useful characteristics to identify cryptic Rhinolophus species. Finally, species delimitation for cryptic species cannot rely on genetic data alone, but such data should be complemented by other evidence, including phenotypic, acoustic data, and geographic distributions to ensure accurate species identification and delineation.

Wildlife-watching and ecotourism are effective approaches for improving public attitudes and raising awareness of wildlife conservation. However, the ability of wildlife tourism to enhance the conservation of less appealing taxa such as bats has rarely been examined. We sampled a total of 197 tourists in Monfort Bat Cave Sanctuary in the Philippines, the world's largest colony of Geoffroy's Rousette (Rousettus amplexicaudatus). Convenient pre/post-visit surveys were conducted to (a) explore the potential of bat-watching to raise tourists’ knowledge, attitudes towards bats, and determine how perceptions vary across demographic classes; and (b) determine potential predictors of conservation willingness among sampled urban tourists. Our study observed an increase in knowledge about bats and 61% of the tourists are willing to support conservation bat protection after the bat-watching visit to the cave site. Tourists’ conservation willingness was associated with age group, prior knowledge of bat ecosystem services, and perceptions about the conservation relevance of bat-watching as a conservation initiative. Our study highlights the effectiveness of short-term engagements such as bat-watching at improving human-bat interactions and suggests such programs should focus on highlighting ecosystem services and benefits of bats. However, it should be noted that we performed this study before the COVID-19 global pandemic and misinformation linking bats to the disease spread has increased since. We expect that public perception will change in the post-COVID-19 period and the conservation willingness survey should be repeated to understand how to counter misconceptions to develop effective bat conservation management in the post-COVID-19.

Humans continue to encroach on the habitats of wild animals, potentially bringing different species into contact that would not typically encounter each other under natural circumstances, and forcing them into stressful, suboptimal conditions. Stressors from unsustainable human land use changes are suspected to dramatically exacerbate the probability of zoonotic spillover of pathogens from their natural reservoir hosts to humans, both by increasing viral load (and shedding) and the interface between wildlife with livestock, pets and humans. Given their known role as reservoir hosts, bats continue to be investigated for their possible role as the origins of many viral outbreaks. However, the participation of bat-associated ectoparasites in the spread of potential pathogens requires further work to establish. Here, we conducted a comprehensive review of viruses, viral genes and other viral sequences obtained from bat ectoparasites from studies over the last four decades. This review summarizes research findings of the seven virus families in which these studies have been performed, including Paramyxoviridae, Reoviridae, Flaviviridae, Peribunyaviridae, Nairoviridae, Rhabdoviridae and Filoviridae. We highlight that bat ectoparasites, including dipterans and ticks, are often found to have medically important viruses and may have a role in the maintenance of these pathogens within bat populations.

This study records for the first time three mammal species as nectar robbers on the ginger Alpinia roxburghii Sweet. We examined the behavior of nectar robbers and compared with earlier studies on a single plant species. We recorded seven species of nectar robbers: three squirrels, one bird, and three bees. Timing of robbing nectars were similar; however, robbing behavior differed among robbers. In particular, squirrels damaged the flower parts while robbing the nectar.