Bat pollinators: a decade of monitoring reveals declining visitation rates for some species in Thailand.

Bats are important pollinators, but they are difficult to study since they are volant and nocturnal. Thus, long-term studies of nectarivorous bats are scarce, despite their potential to help assess trends in bat populations and their pollination services. We used capture rates of nectarivorous bats at chiropterophilous flowers in order to examine temporal trends in bat visitation in an area that is undergoing extensive land use change. We mist-netted at five bat-pollinated plant taxa (Durio zibethinus, Musa acuminata, Oroxylum indicum, Parkia speciosa, and Sonneratia spp.) in southern Thailand over six years between 2011 and 2021. We found that the most common bat species, Eonycteris spelaea, was the main visitor at all five plant taxa and had consistent visitation rates across all study years. In contrast, two other important pollinators, Macroglossus minimus and M. sobrinus, showed 80% declines in the number of individuals netted at mangrove apple (Sonneratia spp.) and banana (Musa acuminata) flowers, respectively. These findings suggest that E. spelaea (a large, cave-roosting species with a broad diet) is more tolerant of anthropogenic change than are Macroglossus bats (small, foliage-roosting species with specialized diets), which may in turn affect the reproductive success of plants pollinated by these species. Our study demonstrates how decade-long monitoring can reveal species-specific temporal patterns in pollinator visitation, emphasizing the need for tailored conservation plans. While the conservation status of most nectarivorous bats in the area is Least Concern, our results indicate that population studies in Southeast Asia are urgently needed for updated bat species conservation assessments.

Complete Genome of Rose Myrtle, Rhodomyrtus tomentosa, and Its Population Genetics in Thai Peninsula.

Several parts of rose myrtle, Rhodomyrtus tomentosa, exhibited profound antibacterial and anti-inflammatory activities, suggesting its potential in healthcare and cosmetics applications. During the past few years, the demand for biologically active compounds in the industrial sectors increased. Therefore, gathering comprehensive information on all aspects of this plant species is essential. Here, the genome sequencing using short and long reads was used to understand the genome biology of R. tomentosa. Inter-simple sequence repeats (ISSR) and simple sequence repeats (SSR) markers, and geometric morphometrics of the leaves of R. tomentosa collected across Thai Peninsula, were determined for population differentiation analysis. The genome size of R. tomentosa was 442 Mb, and the divergence time between R. tomentosa and Rhodamnia argentea, the white myrtle of eastern Australia, was around 15 million years. No population structure was observed between R. tomentosa on the eastern and western sides of the Thai Peninsula using the ISSR and SSR markers. However, significant differences in leaf size and shape of R. tomentosa were observed in all locations.

Nipah virus circulation at human-bat interfaces, Cambodia.

OBJECTIVE: To better understand the potential risks of Nipah virus emergence in Cambodia by studying different components of the interface between humans and bats. METHODS: From 2012 to 2016, we conducted a study at two sites in Kandal and Battambang provinces where fruit bats (Pteropus lylei) roost. We combined research on: bat ecology (reproductive phenology, population dynamics and diet); human practices and perceptions (ethnographic research and a knowledge, attitude and practice study); and Nipah virus circulation in bat and human populations (virus monitoring in bat urine and anti-Nipah-virus antibody detection in human serum). FINDINGS: Our results confirmed circulation of Nipah virus in fruit bats (28 of 3930 urine samples positive by polymerase chain reaction testing). We identified clear potential routes for virus transmission to humans through local practices, including fruit consumed by bats and harvested by humans when Nipah virus is circulating, and palm juice production. Nevertheless, in the serological survey of 418 potentially exposed people, none of them were seropositive to Nipah virus. Differences in agricultural practices among the regions where Nipah virus has emerged may explain the situation in Cambodia and point to actions to limit the risks of virus transmission to humans. CONCLUSION: Human practices are key to understanding transmission risks associated with emerging infectious diseases. Social science disciplines such as anthropology need to be integrated in health programmes targeting emerging infectious diseases. As bats are hosts of major zoonotic pathogens, such integrated studies would likely also help to reduce the risk of emergence of other bat-borne diseases.

A new cost-effective and fast direct PCR protocol for insects based on PBS buffer.

Insect DNA barcoding is a species identification technique used in biodiversity assessment and ecological studies. However, DNA extraction can result in the loss of up to 70% of DNA. Recent research has reported that direct PCR can overcome this issue. However, the success rates could still be improved, and tissues used for direct PCR could not be reused for further genetic studies. Here, we developed a direct PCR workflow that incorporates a 2-min sample preparation in PBS-buffer step for fast and effective universal insect species identification. The developed protocol achieved 100% success rates for amplification in six orders: Mantodea, Phasmatodea, Neuroptera, Odonata, Blattodea and Orthoptera. High and moderate success rates were obtained for five other species: Lepidoptera (97.3%), Coleoptera (93.8%), Diptera (90.5%), Hemiptera (81.8%) and Hymenoptera (75.0%). High-quality sequencing data were also obtained from these amplifiable products, allowing confidence in species identification. The method was sensitive down to 1/4th of a 1-mm fragment of leg or body and its success rates with oven-dried, ethanol-preserved, food, bat guano and museum specimens were 100%, 98.6%, 90.0%, 84.0% and 30.0%, respectively. In addition, the pre-PCR solution (PBS with insect tissues) could be used for further DNA extraction if needed. The workflow will be beneficial in the fields of insect taxonomy and ecological studies due to its low cost, simplicity and applicability to highly degraded specimens.

Habitat and landscape factors influence pollinators in a tropical megacity, Bangkok, Thailand.

BACKGROUND: Pollinators are well known for the ecosystem services they provide, and while urban areas are generally perceived as low-quality habitat for most wildlife, these cities often support a surprising degree of pollinator diversity. The current rapid growth of urban areas and concern over global pollinator declines have spurred numerous studies examining pollinator communities in temperate cities, but knowledge about tropical urban pollinators remains scarce. METHODS: This study investigated the effects of habitat and landscape factors on pollinator richness and abundance in a highly-populated, tropical city: Bangkok, Thailand. We conducted pollinator observations in 52 green areas throughout the city and collected data on patch size, floral abundance, plant richness, location type, and percent vegetation at five spatial scales. RESULTS: Of the 18,793 pollinators observed, over 98% were bees. Both patch size and floral abundance generally had positive effects on pollinators, although there was a significant interaction between the two factors; these findings were generally consistent across all focal taxa (Tetragonula stingless bees, Apis honey bees, Xylocopa carpenter bees, and butterflies). DISCUSSION: Our results demonstrate the importance of maintaining large green areas in cities, since small green areas supported few pollinators, even when floral resources were abundant. Moreover, most pollinator taxa utilized a variety of location types (e.g., public parks, school campuses, temple grounds), with the exception of butterflies, which preferred parks. Our findings are generally consistent with those of temperate urban studies, but additional studies in the tropics are needed before global patterns can be assessed.

Pollination by the locally endangered island flying fox (Pteropus hypomelanus) enhances fruit production of the economically important durian (Durio zibethinus).

Fruit bats provide valuable pollination services to humans through a unique coevolutionary relationship with chiropterophilous plants. However, chiropterophily in the Old World and the pollination roles of large bats, such as flying foxes (Pteropus spp., Acerodon spp., Desmalopex spp.), are still poorly understood and require further elucidation. Efforts to protect these bats have been hampered by a lack of basic quantitative information on their role as ecosystem service providers. Here, we investigate the role of the locally endangered island flying fox Pteropus hypomelanus in the pollination ecology of durian (Durio zibethinus), an economically important crop in Southeast Asia. On Tioman Island, Peninsular Malaysia, we deployed 19 stations of paired infrared camera and video traps across varying heights at four individual flowering trees in a durian orchard. We detected at least nine species of animal visitors, but only bats had mutualistic interactions with durian flowers. There was a clear vertical stratification in the feeding niches of flying foxes and nectar bats, with flying foxes feeding at greater heights in the trees. Flying foxes had a positive effect on mature fruit set and therefore serve as important pollinators for durian trees. As such, semi-wild durian trees-particularly tall ones-may be dependent on flying foxes for enhancing reproductive success. Our study is the first to quantify the role of flying foxes in durian pollination, demonstrating that these giant fruit bats may have far more important ecological, evolutionary, and economic roles than previously thought. This has important implications and can aid efforts to promote flying fox conservation, especially in Southeast Asian countries.

Simulated bacterial infection disrupts the circadian fluctuation of immune cells in wrinkle-lipped bats (Chaerephon plicatus).

BACKGROUND: Leukocyte concentrations follow a circadian pattern in mammals, with elevated values at times of potential contact with pathogens and parasites. We hypothesized that this pattern is disturbed after an immune challenge. METHODS: In Thailand, we captured wrinkle-lipped bats (Chaerephon plicatus), when they returned to their colony at dawn. We challenged half of the animals (experimental group) with bacterial lipopolysaccharides and treated the others only with the carrier liquid (control group). We then compared body mass changes and differences in circulating immune cell counts at 8 h post-treatment. RESULTS: In experimental animals, we observed an increase in total leukocyte and neutrophil numbers of 17% and 95%, respectively. In control animals, concentrations of leukocytes decreased by 44% and those of neutrophils remained constant. Experimental treatment had no effect on lymphocytes, yet changes in eosinophil numbers were explained by sex. Eosinophils decreased by 66% in females and by 62% in males. Basophils and monocytes were rarest among all observed cell types and analysis was either impossible because of low numbers or yielded no significant effects, respectively. DISCUSSION: Our findings show that a simulated bacterial infection triggered a neutrophil-associated immune response in wrinkle-lipped bats, indicating a disruption of the diurnal fluctuation of immune cells. Our study suggests that bats exhibit circadian rhythms in immune cell counts. The magnitude of these fluctuations may vary across species according to specific-specific infection risks associated with colony sizes or specific roosting habits.

Furosemide suppresses ileal and colonic contractility via interactions with GABA-A receptor in mice.

The loop diuretic furosemide has an action to inhibit Na+ -K+ -2Cl- co-transporter at the thick ascending limb of Henle's loop resulting in diuresis. Furosemide also has the non-diuretic effects by binding to GABA-A receptor which may involve the gastrointestinal tract. The aim of this study was to investigate the effects of furosemide on smooth muscle contractions in mice ileum and proximal colon. Each intestinal segment suspended in an organ bath was connected to a force transducer. Signal output of mechanical activity was amplified and recorded for analysis using PowerLab System. After equilibration, the intestine was directly exposed to furosemide, GABA, GABA-A receptor agonist (muscimol), or muscarinic receptor antagonist (atropine). Furosemide (50, 100 and 500 µmol L-1 ) acutely reduced the amplitude of ileal and colonic contraction. In the ileum, 1 mmol L-1 GABA and 10-60 µmol L-1 muscimol significantly increased the amplitude, whereas in the colon, 50-100 mmol L-1 GABA and 60 µmol L-1 muscimol decreased the contractions. The contractions were also significantly suppressed by atropine. To investigate the mechanisms underlying the inhibiting effect of furosemide, furosemide was added to the organ bath prior to the addition of muscimol or atropine. A comparison of furosemide combined with muscimol or atropine group and furosemide group showed no significant difference of the ileal contraction, but the amplitude of colonic contraction significantly decreased when compared to adding furosemide alone. These results suggest that furosemide can reduce the ileal and proximal colonic contraction mediated by blocking and supporting of GABA-A receptor, respectively, resulting in decreased acetylcholine release.

The Diet of the Cave Nectar Bat (Eonycteris spelaea Dobson) Suggests it Pollinates Economically and Ecologically Significant Plants in Southern Cambodia.

Hoem Thavry, Julien Cappelle, Sara Bumrungsri, Lim Thona, and Neil M. Furey (2017) The importance of the cave nectar bat Eonycteris spelaea as a pollinator of economically significant crops and ecologically important plant species is increasingly documented, although information on the plants visited by this widely distributed bat species is currently confined to Thailand and Peninsular Malaysia. We undertook a dietary study on E. spelaea by sampling faecal rain produced by a colony in Kampot, southern Cambodia each month for one year and identifying plant taxa visited by the bats by their pollen. Our results indicate the diet of E. spelaea in Cambodia includes at least 13 plant taxa, eight of which were identi ed to genus or species. Pollen of Sonneratia spp. and Musa spp. had the highest mean monthly frequency at 30.9% and 16.9% respectively, followed by Oroxylum indicum (11.3%), Bombax anceps (11.2%), Parkia spp. (9.8%), Durio zibethinus (6.3%), Ceiba pentandra (6.0%) and Eucalyptus spp. (0.3%). With one exception, all of the plant taxa recorded at our study site are also visited by the bat species in Peninsular Malaysia and Thailand, although their relative dietary contributions differ. This variation likely reflects local differences in the availability, proximity and flowering phenology of chiropterophilous plants between regions, but also suggests a reliance of Cambodian bats on species that ower continuously, coupled with periodic shifts to species that ower profusely for short periods. Only three signi cant colonies (> 1,000 bats) of cave-roosting pteropodids are currently known in Cambodia, all of which are in Kampot and threatened by bushmeat hunting and roost disturbance. We recommend public education and law enforcement e orts to conserve these colonies, not least because Kampot is the premier region for Cambodian durian and this crop depends on nectarivorous bats for fruit set. Protection of mangroves would also bene t durian farmers because these are an important resource for nectarivorous bat populations.

Bird and bat predation services in tropical forests and agroforestry landscapes.

Understanding distribution patterns and multitrophic interactions is critical for managing bat- and bird-mediated ecosystem services such as the suppression of pest and non-pest arthropods. Despite the ecological and economic importance of bats and birds in tropical forests, agroforestry systems, and agricultural systems mixed with natural forest, a systematic review of their impact is still missing. A growing number of bird and bat exclosure experiments has improved our knowledge allowing new conclusions regarding their roles in food webs and associated ecosystem services. Here, we review the distribution patterns of insectivorous birds and bats, their local and landscape drivers, and their effects on trophic cascades in tropical ecosystems. We report that for birds but not bats community composition and relative importance of functional groups changes conspicuously from forests to habitats including both agricultural areas and forests, here termed 'forest-agri' habitats, with reduced representation of insectivores in the latter. In contrast to previous theory regarding trophic cascade strength, we find that birds and bats reduce the density and biomass of arthropods in the tropics with effect sizes similar to those in temperate and boreal communities. The relative importance of birds versus bats in regulating pest abundances varies with season, geography and management. Birds and bats may even suppress tropical arthropod outbreaks, although positive effects on plant growth are not always reported. As both bats and birds are major agents of pest suppression, a better understanding of the local and landscape factors driving the variability of their impact is needed.

Characterization of in utero valproic acid mouse model of autism by local field potential in the hippocampus and the olfactory bulb.

Valproic acid (VPA) mouse model of autism spectrum disorder (ASD) has been characterized mostly by impaired ultrasonic vocalization, poor sociability and increased repetitive self-grooming behavior. However, its neural signaling remained unknown. This study investigated the local field potentials (LFPs) in the dorsal hippocampal CA1 and the olfactory bulb while animals exploring a novel open field. VPA was administered at gestational day 13. The results demonstrated three core features of ASD in male offspring. However, there was no difference in Y-maze performance and locomotor activity. Analysis of hippocampal LFP power revealed significantly increased slow wave (1-4 Hz) and high gamma (80-140 Hz) oscillations and decreased theta (4-12 Hz) activity in VPA mice. In the olfactory bulb, VPA animals showed greater slow wave (1-4 Hz) and beta (25-40 Hz) activity and lower activity of low gamma (55-80 Hz) wave. Regression analysis revealed positive correlations between hippocampal theta power and locomotor speed for both control and VPA-exposed mice. There was no significant difference between groups for modulation index of theta (4-12 Hz) phase modulated gamma (30-200 Hz) amplitude. These findings characterized VPA mouse model with LFP oscillations that might provide better understanding of neural processing in ASD.

Taxonomic implications of geographical variation in Rhinolophus affinis (Chiroptera: Rhinolophidae) in mainland Southeast Asia.

BACKGROUND: Rhinolophusaffinis sensu lato isdistributed throughout Southeast Asia. The taxonomic status of forms attributed to the species is unclear due to the limited sample size with incomplete datasets and the taxa have high variation in morphology and echolocation call frequency. The aim of the study was to evaluate the distribution and taxonomic status of the subspecific forms of R. affinis in mainland Southeast Asia using large sample size with multiple datasets, including morphological, acoustic, and genetic data, both to elucidate taxonomic relationships and to test for congruence between these datasets. RESULTS: Three morphological forms were confirmed within the region; two concur with previously recognized taxa, namely R.affinis macrurus andR.affinis superans,and are strongly supported by morphological and genetic data. The third form is morphologically distinct, but its taxonomic status remains unclear. It is probable that this third form represents a distinct taxonomic entity; however, more data are required to confirm this. R. a. macrurus is known from the north of peninsular Thailand, Cambodia, Myanmar, Laos, and Vietnam (Indochinese subregion); R. a. superans is found throughout the Thai-Malay Peninsula (Sundaic subregion); whilst the third form is presently known from east central Myanmar (Shan state) and lower northern Vietnam (Nghe An Province). CONCLUSIONS: Our results suggest that at least three morphological forms occur in mainland Southeast Asia including one form which appears to be new to science. Echolocation call data for R. affinis are not a robust taxonomic tool as it shows a significant degree of variation which is not explained or supported by genetic and morphological findings. This study highlights significant levels of morphological variation in mainland Southeast Asia and provides an essential basis for further studies aiming to understand the population genetics, phylogeography, and taxonomy of the species.

Nonecholocating fruit bats produce biosonar clicks with their wings.

Because evolution mostly acts over millions of years, the intermediate steps leading to a functional sensory system remain enigmatic. Accordingly, there is an ongoing debate regarding the evolution of bat echolocation. In search of the origin of bat echolocation, we studied how Old World fruit bats, which have always been classified as nonecholocating, orient in complete darkness. We found that two of these nonecholocating species used click-like sounds to detect and discriminate objects in complete darkness. However, we discovered that this click-based echo sensing is rudimentary and does not allow these bats to estimate distance accurately as all other echolocating bats can. Moreover, unlike all other echolocating bats, which generate pulses using the larynx or the tongue, these bats generated clicks with their wings. We provide evidence suggesting that all Old World fruit bats can click with their wings. Although this click-based echo sensing used by Old World fruit bats may not represent the ancestral form of current (laryngeal) bat echolocation, we argue that clicking fruit bats could be considered behavioral fossils, opening a window to study the evolution of echolocation.

Parasite and viral species richness of Southeast Asian bats: Fragmentation of area distribution matters.

Interest in bat-borne diseases and parasites has grown in the past decade over concerns for human health. However, the drivers of parasite diversity among bat host species are understudied as are the links between parasite richness and emerging risks. Thus, we aimed at exploring factors that explain macro and microparasite species richness in bats from Southeast Asia, a hotspot of emerging infectious diseases. First, we identified bat species that need increased sampling effort for pathogen discovery. Our approach highlights pathogen investigation disparities among species within the same genus, such as Rhinolophus and Pteropus. Secondly, comparative analysis using independent contrasts method allowed the identification of likely factors explaining parasite and viral diversity of bats. Our results showed a key role of bat distribution shape, an index of the fragmentation of bat distribution, on parasite diversity, linked to a decrease for both viral and endoparasite species richness. We discuss how our study may contribute to a better understanding of the link between parasite species richness and emergence.

Detection of environmental sources of Histoplasma capsulatum in Chiang Mai, Thailand, by nested PCR.

Histoplasmosis is a systemic mycosis caused by inhaling spores of Histoplasma capsulatum, a dimorphic fungus. This fungus grows in soil contaminated with bat and avian excreta. Each year, patients with disseminated histoplasmosis have been diagnosed in Chiang Mai, northern Thailand. No published information is currently available on the environmental sources of this fungus in Chiang Mai or anywhere else in Thailand. The aim of this study was to detect H. capsulatum in soil samples contaminated with bat guano and avian droppings by nested PCR. Two hundred and sixty-five samples were collected from the following three sources: soil contaminated with bat guano, 88 samples; soil contaminated with bird droppings, 86 samples; and soil contaminated with chicken droppings, 91 samples. Genomic DNA was directly extracted from each sample, and H. capsulatum was detected by nested PCR using a primer set specific to a gene encoding 100-kDa-like protein (HcI, HcII and HcIII, HcIV). Histoplasma capsulatum was detected in seven of 88 soil samples contaminated with bat guano, one of 21 soil samples contaminated with pigeon droppings and 10 of 91 soil samples contaminated with chicken droppings. The results indicate the possibility of the association of bat guano and chicken droppings with H. capsulatum in this area of Thailand.

Near-complete extinction of native small mammal fauna 25 years after forest fragmentation.

Tropical forests continue to be felled and fragmented around the world. A key question is how rapidly species disappear from forest fragments and how quickly humans must restore forest connectivity to minimize extinctions. We surveyed small mammals on forest islands in Chiew Larn Reservoir in Thailand 5 to 7 and 25 to 26 years after isolation and observed the near-total loss of native small mammals within 5 years from <10-hectare (ha) fragments and within 25 years from 10- to 56-ha fragments. Based on our results, we developed an island biogeographic model and estimated mean extinction half-life (50% of resident species disappearing) to be 13.9 years. These catastrophic extinctions were probably partly driven by an invasive rat species; such biotic invasions are becoming increasingly common in human-modified landscapes. Our results are thus particularly relevant to other fragmented forest landscapes and suggest that small fragments are potentially even more vulnerable to biodiversity loss than previously thought.

Rediscovery of Biswamoyopterus (Mammalia: Rodentia: Sciuridae: Pteromyini) in Asia, with the description of a new species from Lao PDR.

A new species of the flying squirrel genus Biswamoyopterus is described from Lao PDR. It is based on a single specimen collected from a local food market at Ban Thongnami, Pak Kading District, Bolikhamxai Province. The new taxon shows close affinities to Biswamoyopterus biswasi, which is only known from the holotype collected in 1981, 1250 km from the current locality, in Arunachal Pradesh, Northeast India. However, it differs substantially in pelage colour, most particularly on the ventral surfaces of the body, patagia, tail membrane, and tail. The single specimen was found in an area of central Lao PDR, which is characterised by its extensive limestone karst formations and which is home to other rare endemic rodents, including the Khanyou (Laonastes aenigmamus) nd the Lao limestone rat (Saxatilomyspaulinae).