Terrestrial Species Research Articles

Metabolic Water As a Route for Water Acquisition in Vertebrates Inhabiting Dehydrating Environments.

Vertebrates have expanded their habitats during evolution, which accompanies diversified routes for water acquisition. Water is acquired by oral intake and subsequent absorption by the intestine in terrestrial and marine animals which are subjected to constant dehydration, whereas most water is gained osmotically across body surfaces in freshwater animals. In addition, a significant amount of water, called metabolic water, is produced within the body by the oxidation of hydrogen in organic substrates. The importance of metabolic water production as a strategy for water acquisition has been well documented in desert animals, but its role has attracted little attention in marine animals which also live in a dehydrating environment. In this article, the author has attempted to reevaluate the role of metabolic water production in body fluid regulation in animals inhabiting desiccating environments. Because of the exceptional ability of their kidney, marine mammals are thought to typically gain water by drinking environmental seawater and excreting excess NaCl in the urine. On the other hand, it is established that marine teleosts drink seawater to enable intestinal water and ion absorption, and the excess NaCl is excreted by branchial ionocytes. In addition to the oral route, we suggest through experiments using eels that water production by lipid metabolism is an additional route for water acquisition when they encounter seawater. It seems that metabolic water production contributes to counteract dehydration before mechanisms for water regulation are reversed from excretion in freshwater to acquisition in seawater.

Rainforest transformation reallocates energy from green to brown food webs

Terrestrial animal biodiversity is increasingly being lost because of land-use change1,2. However, functional and energetic consequences aboveground and belowground and across trophic levels in megadiverse tropical ecosystems remain largely unknown. To fill this gap, we assessed changes in energy fluxes across ‘green’ aboveground (canopy arthropods and birds) and ‘brown’ belowground (soil arthropods and earthworms) animal food webs in tropical rainforests and plantations in Sumatra, Indonesia. Our results showed that most of the energy in rainforests is channelled to the belowground animal food web. Oil palm and rubber plantations had similar or, in the case of rubber agroforest, higher total animal energy fluxes compared to rainforest but the key energetic nodes were distinctly different: in rainforest more than 90% of the total animal energy flux was channelled by arthropods in soil and canopy, whereas in plantations more than 50% of the energy was allocated to annelids (earthworms). Land-use change led to a consistent decline in multitrophic energy flux aboveground, whereas belowground food webs responded with reduced energy flux to higher trophic levels, down to −90%, and with shifts from slow (fungal) to fast (bacterial) energy channels and from faeces production towards consumption of soil organic matter. This coincides with previously reported soil carbon stock depletion3. Here we show that well-documented animal biodiversity declines with tropical land-use change4–6 are associated with vast energetic and functional restructuring in food webs across aboveground and belowground ecosystem compartments.

What`s crawling in the dark? An annotated list of gastropods in Greek caves

Greece is covered by a high percentage of carbonate rocks, resulting in a significant number of over 10,000 caves within its territory. Their fauna is not well known. Concerning gastropods, 68 terrestrial species have been reported from 70 caves. In this study, we contribute to the knowledge of the snails inhabiting Greek caves. We analyzed the literature and studied all the material deposited in the Natural History Museum of Crete, to create a first comprehensive list of gastropods of Greek caves. The number of caves from which gastropod species were reported increased to 182, while the number of known terrestrial species increased to 113. Fourteen of these species live only in caves, twelve of which are endemic of Greece. However, the fauna of Greek caves is still understudied. With the increasing exploration of Greek caves, the number of the known cave-dwelling gastropod species will increase and new species will be discovered.

KELEA (Kinetic Energy Limiting Electrostatic Attraction) stimulated Alternative Cellular Energy (ACE) pathway can restore allostasis in heavily polluted collections of water with secondary benefits to birds and land animals

KELEA (Kinetic Energy Limiting Electrostatic Attraction) stimulated Alternative Cellular Energy (ACE) pathway can restore allostasis in heavily polluted collections of water with secondary benefits to birds and land animals

Regeneration Abilities among Extant Animals Depend on Their Evolutionary History and Life Cycles.

The present brief manuscript summarizes the main points supporting recently proposed hypotheses explaining the different distributions of regenerative capacity among invertebrates and vertebrates. The new hypotheses are based on the evolution of regeneration from marine animals to the terrestrial animals derived from them. These speculations suggest that animals that were initially capable of broad regeneration in the sea underwent epigenetic modifications during terrestrial adaptation that determined the loss of their regenerative abilities in sub-aerial conditions. These changes derived from the requirements of life on land that include variable dry and UV-exposed conditions. Terrestrial conditions do not allow for organ regeneration, especially in arthropods and amniotes. Nematodes, the other main metazoan group unable of regeneration, instead evolved eutely (a fixed number of body cells), a process which is incompatible with regeneration. All these changes involved gene loss, modification and new gene interactions within the genomes of terrestrial adapting animals that gave rise to sophisticated invertebrates and vertebrates adapted to living on land but with low cellular plasticity.

Recumbirostran ‘microsaurs’ are not amniotes

Amniota is a tetrapod clade that includes extant mammals and reptiles, including birds, as well as a wealth of extinct taxa extending over 318 million years of Earth’s history. For over three decades, Amniota has been treated as a crown group, but the content of stem amniotes has varied among large-scale analyses of Palaeozoic tetrapods. Nine recent phylogenetic studies on Palaeozoic tetrapods, all derived from each other, have potentially changed our perspectives on amniote origins and early evolution. The first of these analyses posited that two particular clades of Palaeozoic tetrapods, Recumbirostra and Lysorophia, both previously considered to be lepospondyl stem-amniotes, are actually amniotes. This raises the possibility that early amniotes included both small fossorial and terrestrial animals, some of which show strong evidence of limb reduction. Here we show that the strange position of these small, often fossorial and limb-reduced tetrapods in Amniota is the result of two fundamental flaws: (1) the phylogenetic analysis combines two separate data matrices that were each designed to test only patterns of relationships among early anamniote tetrapods; and (2) the absence of key amniote characters. Our reanalysis of Palaeozoic tetrapod interrelationships reinstates recumbirostrans and lysorophians outside Amniota. The results of our study impact other recent studies that have simply accepted the codings in the data matrix that resulted in the hypothesis that recumbirostrans are amniotes and raises important questions about the initial diversification of amniotes and the evolution of fossoriality and limb reduction among tetrapods.

Inventory of Orchid Species in Customary Forest Area of Cipta Karya Village, Sungai Betung District, Bengkayang Regency

Orchids are one of the flowering plants that have many species and are widely distributed, as well as having different attractions and characteristics in each species and habitat. The aim of this research to inventory the types of orchids in Customary Forest Area of Cipta Karya Village, Bengkayang Regency. The research was conducted using the cruise method. The characteristics observed were the morphology of orchids such as roots, pseudobulbs, stems, leaves, and flowers. The inventory results found 13 species of orchids. Epiphytic orchids were found more commonly in this area, with a total of 9 species, while terrestrial orchids were found with 4 species. The epiphytic orchid species foundware composed of Appendicula sp., Bulbophyllum sp., Coelogyne speciosa, Cymbidium sp., Dendrobium crumenatum, Dendrobium anosmum, Luisia sp., Micropera pallida and Oberonia sp. The terrestrial orchid species found ware composed of Acanthephippium javanicum, Malaxis latifolia, Nephelaphyllum pulchrum, and Spathoglottis plicata.

Temporal and spatial refugia modify predation risk for non-native crabs in rocky intertidal habitats.

Populations of the non-native Asian shore crab, Hemigrapsus sanguineus, now dominate the rocky intertidal of southern New England, USA. High abundances suggest the recent invader may have experienced enhanced success as a result of enemy release. While larvae and juveniles may serve as a food source for ecologically important species, little is known about predation of mature H. sanguineus or the influence of habitat on predation pressure. To assess natural predation rates of adult H. sanguineus, crabs were tethered in the intertidal at Clarks Cove in New Bedford, MA. Crabs were left in situ for half of a tidal cycle then observed for signs of predation. Results of separate high and low tide trials showed that adult crabs were preyed upon at both high and low tide, though at a significantly higher rate during high tide during both daytime and nighttime, suggesting predation by aquatic species is greater than that by terrestrial species. To investigate the role of habitat as refuge from predation, a laboratory experiment manipulated the complexity of habitat provided to crabs in the presence of a native fish predator. Results indicate better refuge is provided by more complex shelter. Together, findings suggest that fish, crabs, and/or diving birds are important predators for H. sanguineus in the invaded range and that habitat refuge acts to reduce predation pressure.

Mangrove species found in contrasting environments show differing phytohormonal responses to variation in soil bulk density

Background and aimsMangrove species respond to variation in soil bulk density (BD). However, very little is known about the regulatory mechanisms that trigger these responses.MethodsEndogenous concentrations of different phytohormones were measured in the roots of two mangrove species (Avicennia marina and Rhizophora stylosa) grown in low and high BD soils. The potential involvement of ethylene in regulating plant growth responses was tested by applying the ethylene biosynthesis inhibitors cobalt chloride (CoCl2) and aminoisobutyric acid (AIB).ResultsThe two mangrove species responded differently to variation in soil BD. High BD decreased root growth of R. stylosa, but not A. marina. Soil BD had no effect on root phytohormone levels in R. stylosa, but loose soils increased 1-aminocyclopropane-1-carboxylic acid whilst decreasing salicylic acid and gibberellin in A. marina. Applying ethylene inhibitors enhanced R. stylosa root growth, while increasing indole-3-acetic acid but decreasing isopentenyl adenine levels. In contrast, AIB inhibited A. marina root growth, while increasing trans-zeatin levels. Ethylene inhibitors affected salicylic acid levels in both species.ConclusionSalicylic acid is central to root growth responses to variation in BD in A. marina. Conversely, the interaction of ethylene and gibberellin drives responses in R. stylosa. Hormonal interactions involving ethylene potentially reflect the adaptations of the two species to differing conditions within the intertidal zone, with A. marina behaving like an aquatic species and R. stylosa behaving like a terrestrial species.

Nutrients from spawning salmon influence leaf area, tissue density, and nitrogen-15 in riparian plant leaves.

Nutrient subsidies have significant impacts on ecosystems by connecting disjunct habitats, often through long-distance animal migrations. Salmon migrations on the North Pacific coasts provide these kinds of nutrient subsidies from senescent fish at the end of their life cycle, which can have significant ecological effects on terrestrial species. This can include impacts on individuals, populations, and communities, where shifts in community composition towards plant species that indicate nitrogen-rich soils have been documented. We investigated the effects of variation in salmon spawning density on the leaf traits of four common riparian plant species on the central coast of British Columbia, Canada. We found that all plant species had higher foliar salmon-derived nitrogen on streams with a higher spawning density. Three of the four species had larger leaves, and one species also had higher leaf mass per area on streams with more salmon. However, we found no differences in leaf greenness or foliar percent nitrogen among our study streams. These results demonstrate that nutrient subsidies from spawning salmon can have significant impacts on the ecology, morphology, and physiology of riparian plants, which lends support to a mechanism by which certain plants are more common on productive salmon streams.

Pathology, microbiology, and genetic diversity associated with Erysipelothrix rhusiopathiae and novel Erysipelothrix spp. infections in southern sea otters (Enhydra lutris nereis).

Erysipelothrix spp., including E. rhusiopathiae, are zoonotic bacterial pathogens that can cause morbidity and mortality in mammals, fish, reptiles, birds, and humans. The southern sea otter (SSO; Enhydra lutris nereis) is a federally-listed threatened species for which infectious disease is a major cause of mortality. We estimated the frequency of detection of these opportunistic pathogens in dead SSOs, described pathology associated with Erysipelothrix infections in SSOs, characterized the genetic diversity and antimicrobial susceptibility of SSO isolates, and evaluated the virulence of two novel Erysipelothrix isolates from SSOs using an in vivo fish model. From 1998 to 2021 Erysipelothrix spp. were isolated from six of >500 necropsied SSOs. Erysipelothrix spp. were isolated in pure culture from three cases, while the other three were mixed cultures. Bacterial septicemia was a primary or contributing cause of death in five of the six cases. Other pathology observed included suppurative lymphadenopathy, fibrinosuppurative arteritis with thrombosis and infarction, bilateral uveitis and endophthalmitis, hypopyon, petechia and ecchymoses, mucosal infarction, and suppurative meningoencephalitis and ventriculitis. Short to long slender Gram-positive or Gram-variable bacterial rods were identified within lesions, alone or with other opportunistic bacteria. All six SSO isolates had the spaA genotype-four isolates clustered with spaA E. rhusiopathiae strains from various terrestrial and marine animal hosts. Two isolates did not cluster with any known Erysipelothrix spp.; whole genome sequencing revealed a novel Erysipelothrix species and a novel E. rhusiopathiae subspecies. We propose the names Erysipelothrix enhydrae sp. nov. and Erysipelothrix rhusiopathiae ohloneorum ssp. nov. respectively. The type strains are E. enhydrae UCD-4322-04 and E. rhusiopathiae ohloneorum UCD-4724-06, respectively. Experimental injection of tiger barbs (Puntigrus tetrazona) resulted in infection and mortality from the two novel Erysipelothrix spp. Antimicrobial susceptibility testing of Erysipelothrix isolates from SSOs shows similar susceptibility profiles to isolates from other terrestrial and aquatic animals. This is the first description of the pathology, microbial characteristics, and genetic diversity of Erysipelothrix isolates recovered from diseased SSOs. Methods presented here can facilitate case recognition, aid characterization of Erysipelothrix isolates, and illustrate assessment of virulence using fish models.

TerrANTALife 1.0 Biodiversity data checklist of known Antarctic terrestrial and freshwater life forms.

Incomplete species inventories for Antarctica represent a key challenge for comprehensive ecological research and conservation in the region. Additionally, data required to understand population dynamics, rates of evolution, spatial ranges, functional traits, physiological tolerances and species interactions, all of which are fundamental to disentangle the different functional elements of Antarctic biodiversity, are mostly missing. However, much of the fauna, flora and microbiota in the emerged ice-free land of the continent have an uncertain presence and/or unresolved status, with entire biodiversity compendia of prokaryotic groups (e.g. bacteria) being missing. All the available biodiversity information requires consolidation, cross-validation, re-assessment and steady systematic inclusion in order to create a robust catalogue of biodiversity for the continent. We compiled, completed and revised eukaryotic species inventories present in terrestrial and freshwater ecosystems in Antarctica in a new living database: terrANTALife (version 1.0). The database includes the first integration in a compendium for many groups of eukaryotic microorganisms. We also introduce a first catalogue of amplicon sequence variants (ASVs) of prokaryotic biodiversity. Available compendia and literature to date were searched for Antarctic terrestrial and freshwater species, integrated, taxonomically harmonised and curated by experts to create comprehensive checklists of Antarctic organisms. The final inventories comprises 470 animal species (including vertebrates, free-living invertebrates and parasites), 306 plants (including all Viridiplantae: embryophytes and green algae), 997 fungal species and 434 protists (sensu lato). We also provide a first account for many groups of microorganisms, including non-lichenised fungi and multiple groups of eukaryotic unicellular species (Stramenophila, Alveolata and Rhizaria (SAR), Chromists and Amoeba), jointly referred to as "protists". In addition, we identify 1753 bacterial (obtained from 348117 ASVs) and 34 archaeal genera (from 1848 ASVs), as well as, at least, 14 virus families. We formulate a basic tree of life in Antarctica with the main lineages listed in the region and their "known-accepted-species" numbers.

Creating a systematic prioritization of stream reaches for conservation of aquatic species

AbstractHuman impacts on aquatic ecosystems have resulted in systemic declines of global freshwater species abundance and richness. Conservation and governmental groups worldwide have designated protected areas to preserve the remaining diversity. The biodiversity hotspot approach, which designates areas based on high levels of species richness, has been useful for identifying areas to protect both terrestrial and aquatic species. However, for freshwater species, additional approaches are warranted to identify specific stream reaches for protection and/or restoration. To address this issue, we present a methodology to create a Gridded River Identification System (GRIS) for river segments based on 30 arc‐second grids (~0.9 km) using the USGS National Hydrography High Resolution Dataset. To demonstrate the utility of this approach, we obtained occurrence data for six imperiled freshwater mussel species in Texas and created ensemble species distribution models (ESDMs) based on climate and topographical variables. Predicted occupancies were overlayed onto the GRIS in Texas. The predicted occupancies were rank ordered from 1 to 5, with 1 being the lowest probability of occupancy and 5 being the greatest. The rank‐ordered segments were then used to identify reaches for conservation and restoration activities. Our approach is novel and widely applicable to other freshwater species so long as distribution information is available. The GRIS can also be easily developed for stream systems outside of the current study area. Future studies could build upon our framework by incorporating additional taxa data and projected changes in climate and land use.

Duration and variability of spring green-up mediate population consequences of climate change.

Single phenological measures, like the average rate of phenological advancement, may be insufficient to explain how climate change is driving trends in animal populations. Here, we develop a multifactorial concept of spring phenology-including the onset of spring, spring duration, interannual variability, and their temporal changes-as a driver for population dynamics of migratory terrestrial species in seasonal environments. Using this conceptual model, we found that effects of advancing spring phenology on animal populations may be buffered or amplified depending on the duration and interannual variability of spring green-up, and those effects are modified by evolutionary and plastic adaptations of species. Furthermore, we compared our modelling results with empirical data on normalized difference vegetation index-based spring green-up phenology and population trends of 106 European landbird finding similar associations. We conclude how phenological changes are expected to affect migratory bird populations across Europe and identify regions that are particularly prone to suffer population declines.

Red vision in animals is broadly associated with lighting environment but not types of visual task.

Red sensitivity is the exception rather than the norm in most animal groups. Among species with red sensitivity, there is substantial variation in the peak wavelength sensitivity (λmax) of the long wavelength sensitive (LWS) photoreceptor. It is unclear whether this variation can be explained by visual tuning to the light environment or to visual tasks such as signalling or foraging. Here, we examine long wavelength sensitivity across a broad range of taxa showing diversity in LWS photoreceptor λmax: insects, crustaceans, arachnids, amphibians, reptiles, fish, sharks and rays. We collated a list of 161 species with physiological evidence for a photoreceptor sensitive to red wavelengths (i.e. λmax ≥ 550 nm) and for each species documented abiotic and biotic factors that may be associated with peak sensitivity of the LWS photoreceptor. We found evidence supporting visual tuning to the light environment: terrestrial species had longer λmax than aquatic species, and of these, species from turbid shallow waters had longer λmax than those from clear or deep waters. Of the terrestrial species, diurnal species had longer λmax than nocturnal species, but we did not detect any differences across terrestrial habitats (closed, intermediate or open). We found no association with proxies for visual tasks such as having red morphological features or utilising flowers or coral reefs. These results support the emerging consensus that, in general, visual systems are broadly adapted to the lighting environment and diverse visual tasks. Links between visual systems and specific visual tasks are commonly reported, but these likely vary among species and do not lead to general patterns across species.

CONSTRAINTS FACING AGROSILVOPASTORAL PRACTICES AMONG HOUSEHOLDS IN OGUN STATE, NIGERIA

Agrosilvopastoral practices are recognized as a link between agriculture, forestry and a sustainable strategy for land management, particularly in tropical and subtropical developing nations. This study examined the limitations of agrosilvopastoral practices among households in Ogun State. A well thought out questionnaire was used to gather information from 240 respondents in total. Descriptive statistics were used to analyze the data collected. Findings revealed that agrosilvopastoral practices were dominated by men, with a mean age and family size of 54 years and 6 persons respectively. More over half (62.25%) of the farmers were literate with mean years of experience in farming being 26years. Result revealed that inadequate capital (1st) is the most serious challenge facing the agrosilvopastoral households, followed by insect infestation (2nd), instability in market price (3rd), inadequate access to land for farming (4th), high cost of transportation (5th), insufficient market centers (6th), source of inputs (7th), technical knowledge (8th), mode of land acquisition (9th) and animal destruction of crops (10th). The study thereby recommends that extension workers from different organizations should educate farmers about agrosilvopastoral practices through seminars, and backing being offered to farmers in the terms of loans, hybrid seeds and credit provision for increased production.

Biodegradation of PVCs through in-vitro identification of Bacillus albus and computational pathway analysis of ABH enzyme.

Microplastics pose significant challenges to ecosystems and organisms. They can be ingested by marine and terrestrial species, leading to potential health risks and ecological disruptions. This study aims to address the urgent need for effective remediation strategies by focusing on the biodegradation of microplastics, specifically polyvinyl chloride (PVC) derivatives, using the bacterial strain Bacillus albus. The study provides a comprehensive background on the accumulation of noxious substances in the environment and the importance of harnessing biodegradation as an eco-friendly method for pollutant elimination. The specific objective is to investigate the enzymatic capabilities of Bacillus albus, particularly the alpha/beta hydrolases (ABH), in degrading microplastics. To achieve this, in-silico studies were conducted, including analysis of the ABH protein sequence and its interaction with potential inhibitors targeting PVC derivatives. Docking scores of -7.2 kcal/mol were obtained to evaluate the efficacy of the interactions. The study demonstrates the promising bioremediation prospects of Bacillus albus for microplastics, highlighting its potential as a key player in addressing microplastic pollution. The findings underscore the urgent need for further experimental validation and practical implementation of Bacillus albus in environmental remediation strategies.

Development and application of a second-generation multilingual tool for invasion risk screening of non-native terrestrial plants

Under the increasing threat to native ecosystems posed by non-native species invasions, there is an urgent need for decision support tools that can more effectively identify non-native species likely to become invasive. As part of the screening (first step) component in non-native species risk analysis, decision support tools have been developed for aquatic and terrestrial organisms. Amongst these tools is the Weed Risk Assessment (WRA) for screening non-native plants. The WRA has provided the foundations for developing the first-generation WRA-type Invasiveness Screening Kit (ISK) tools applicable to a range of aquatic species, and more recently for the second-generation ISK tools applicable to all aquatic organisms (including plants) and terrestrial animals. Given the most extensive usage of the latter toolkits, this study describes the development and application of the Terrestrial Plant Species Invasiveness Screening Kit (TPS-ISK). As a second-generation ISK tool, the TPS-ISK is a multilingual turnkey application that provides several advantages relative to the WRA: (i) compliance with the minimum standards against which a protocol should be evaluated for invasion process and management approaches; (ii) enhanced questionnaire comprehensiveness including a climate change component; (iii) provision of a level of confidence; (iv) error-free computation of risk scores; (v) multilingual support; (vi) possibility for across-study comparisons of screening outcomes; (vii) a powerful graphical user interface; (viii) seamless software deployment and accessibility with improved data exchange. The TPS-ISK successfully risk-ranked five representative sample species for the main taxonomic groups supported by the tool and ten angiosperms previously screened with the WRA for Turkey. The almost 20-year continuous development and evolution of the ISK tools, as opposed to the WRA, closely meet the increasing demand by scientists and decision-makers for a reliable, comprehensive, updatable and easily deployable decision support tool. For terrestrial plant screening, these requirements are therefore met by the newly developed TPS-ISK.

Sources of bias in applying close-kin mark-recapture to terrestrial game species with different life histories.

Close-kin mark-recapture (CKMR) is a method analogous to traditional mark-recapture but without requiring recapture of individuals. Instead, multilocus genotypes (genetic marks) are used to identify related individuals in one or more sampling occasions, which enables the opportunistic use of samples from harvested wildlife. To apply the method accurately, it is important to build appropriate CKMR models that do not violate assumptions linked to the species' and population's biology and sampling methods. In this study, we evaluated the implications of fitting overly simplistic CKMR models to populations with complex reproductive success dynamics or selective sampling. We used forward-in-time, individual-based simulations to evaluate the accuracy and precision of CKMR abundance and survival estimates in species with different longevities, mating systems, and sampling strategies. Simulated populations approximated a range of life histories among game species of North America with lethal sampling to evaluate the potential of using harvested samples to estimate population size. Our simulations show that CKMR can yield nontrivial biases in both survival and abundance estimates, unless influential life history traits and selective sampling are explicitly accounted for in the modeling framework. The number of kin pairs observed in the sample, in combination with the type of kinship used in the model (parent-offspring pairs and/or half-sibling pairs), can affect the precision and/or accuracy of the estimates. CKMR is a promising method that will likely see an increasing number of applications in the field as costs of genetic analysis continue to decline. Our work highlights the importance of applying population-specific CKMR models that consider relevant demographic parameters, individual covariates, and the protocol through which individuals were sampled.

Identifying umbrella and indicator species to support multispecies population connectivity in a Himalayan biodiversity hotspot

Successful conservation hinges on the reliable prediction and prioritisation of population core habitats and dispersal corridors. However, reliance on single-species connectivity has produced mixed results and, in most cases, failed to protect other species under the appealing but often unfulfilled guise of umbrella protection. The main objectives of this study were: (1) to identify core habitats and dispersal corridors for population connectivity of 15 terrestrial mammal species across Bhutan, (2) to identify indicator species of wildlife connectivity, (3) to prioritise core areas and dispersal corridors for the focal umbrella and indicator species, (4) to develop composite multispecies connectivity maps and evaluate the effectiveness of the existing protected area network. We used kernel density (KD) to identify core habitat areas and factorial least-cost path (LCP) to map dispersal corridors. We used correlation and cluster analyses to identify umbrella and indicator species, and quantitative ranking to prioritise core areas and corridors for these focal species. Finally, we produced maps of multispecies core areas and corridors, and identified gaps in the existing protected area network (PAN). We found that ungulates were generally more effective than carnivores in serving as umbrella species for core habitat protection, with Asian elephant (Elephas maximus) emerging as the most effective umbrella for core habitats and muntjac (Muntiacus muntjac) for dispersal corridors. We identified six focal connectivity indicator species in Bhutan: tiger (Panthera tigris), dhole (Cuon alpinus), clouded leopard (Neofelis nebulosa), Asiatic black bear (Ursus thibetanus), Asian elephant, and sambar deer (Rusa unicolor). Most indicator species’ core areas and dispersal corridors outside the PAN were in southern Bhutan, indicating opportunities for increased multispecies connectivity through expanding protected area network in this region. A total of 40% of multispecies core areas and 24% of dispersal corridors were under PAN, with large parts of Bhutan offering suitable core areas and corridors for multispecies conservation. Core areas and corridors of the 15 considered species only partially overlapped, stressing the importance of multi-species consideration in designating wildlife corridors.