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Understanding human–canid conflict and coexistence must focus on documenting human–canid interactions and identifying the underlying drivers of reciprocal human attitude which enables appropriate strategies to minimize conflict and forge coexistence. The dhole (Cuon alpinus), Asia's most widely distributed wild canid, is highly threatened by human persecution and anthropogenic activities. Despite its “endangered” status, its ecological role as an apex predator, negative interactions with humans, and dhole-specific attitude studies are limited, thus hindering the development of a comprehensive dhole-conservation strategy. Here, we investigate the influence of socioeconomic factors of age, gender, income, residency inside/outside a protected area (PA), and other variables (cultural beliefs, livestock loss, and quantity of livestock loss) on the attitudes of local people and support for dhole conservation in the Himalayan Kingdom of Bhutan. We conducted a semi-structured questionnaire survey of 1,444 households located within the PA and non-PA from four representative regions in the country. Using R programming, we ran Pearson's chi-square test of independence to test the overall difference in the attitude and support for dhole conservation, followed by recursive partitioning through a conditional inference regression tree to identify its significant covariates with the highest explanatory power. Majority (79.1%) of respondents (χ2 = 488.6; df = 1; p < 0.001) disliked the dhole over those who liked it. More than half (57.7%) (χ2 = 412.7; df = 2; p < 0.001) opposed dhole conservation over those who either supported or remained neutral. Experience of livestock loss to dholes was the primary ( p < 0.001) factor influencing the negative attitude and opposition to dhole conservation, despite an acknowledgment of the ecological role of the dhole in controlling agricultural crop predators. Our study, which is the first-ever survey in Bhutan, solely focused on investigating human attitudes and perceptions toward the dhole, indicating that livestock loss to dholes transcends all positive attitudes to the species and drives a predominant dislike and opposition to its conservation. To improve the attitude and support toward the dhole and to foster dhole–human coexistence, livestock predation by dholes needs alleviation by improving the existing animal husbandry, in conjunction with promoting conservation awareness on this species.

Most canids face population declines and range contractions worldwide. Although the dhole (Cuon alpinus) is widely distributed across 10 countries in South and Southeast Asia, limited studies exist on this species. Despite its globally “Endangered” status and ecological role as an apex predator, assessments on its distribution are limited to a few landscapes and countries. This explains the lack of a dhole-specific species conservation plan in most range countries, including Bhutan where no current population estimate exists. The species has also recovered from a country-wide poisoning campaign in the 1970s and 80s. In this study, we determine the dhole's distribution pattern and assess the protection and connectivity of dhole habitat in Bhutan. We anticipated dholes to be extant within their habitat well-represented in protected areas (PAs) and biological corridors (BCs). We used 721 georeferenced dhole occurrence records and eight environmental variables in MaxEnt software to model potential dhole distribution and habitat suitability. The model output was overlaid on the spatial layers of PAs and BCs to assess habitat protection and connectivity. As anticipated, we found the dhole widely distributed in all districts, PAs, and BCs in Bhutan. Dholes were recorded at the highest elevation range limit of 4,980 m above sea level, which overlapped with the “Vulnerable” snow leopard (Panthera uncia). Our model identified 72% (27,634 km2) of the country as suitable areas for dholes, of which, 31% (11,899 km2) was highly suitable and 41% (15,735 km2) was moderately suitable. Contrary to our expectation, PAs and BCs encompassed only 29% (8,046 km2) and 12% (3,185 km2) of suitable areas for dholes, respectively. A vast majority of the areas we deemed suitable for dholes currently remain unprotected, thus making dholes more vulnerable to human persecution and local extermination. We recommend adjusting PA boundaries to fully encompass suitable dhole habitat, and also advocate improved livestock husbandry to reduce dhole related livestock predation and minimize conflict, thereby ensuring its long-term survival in Bhutan.

Abstract Protected area networks (PAN) are essential for conserving wide‐ranging apex predators but their adequacy in species protection has rarely been assessed. Here, we assess the adequacy of Bhutan's PAN in conserving and providing connectivity to the endangered tiger (Panthera tigris). We determine the current extent of tiger habitat, predict new suitable habitat, identify potential corridors, and empirically estimate the range of tiger numbers that Bhutan can spatially support. We use two spatial models with different approaches to ascertain current tiger distribution and predict new suitable tiger areas: (a) an expert model based on tiger ecology and (b) an observation model from observed tiger distribution. The expert model identified more suitable tiger areas (32,887 km2) over the observation model (29,962 km2), with the PAN encompassing 46% and 45% of predicted suitable areas, respectively. Vast suitable tiger habitat remains unprotected. Based on our estimates of total suitable habitats, Bhutan can spatially support 138–151 tigers compared to the current estimate of 103, thereby precluding a doubling in tiger numbers. To ensure adequate protection of tigers in Bhutan, we recommend readjusting and/or expanding existing PAN boundaries, including the designation of new corridors, protecting habitats, and conserving prey populations.

The Bhutan Takin Budorcas whitei Lydekker, 1907 is endemic to Bhutan and it is categorized as Vulnerable by the IUCN Red List of Threatened Species. While the other Takin species have been studied in China (Golden Takin B. bedfordi; Sichuan Takin B. tibetana) and India (Mishmi Takin B. taxicolor), only one study has focused on the Bhutan Takin. In this paper, we report the current distribution and conservation status of the Bhutan Takin using the information gathered through field surveys, interviews and unpublished reports. Bhutan Takin are seasonal migrants, occurring between 1500–5550 m, preferring areas in close proximity to river valleys and geothermal outlets (hot springs). Takin avoid areas that are disturbed by road construction and power transmission lines, and where they have to compete for forage with domestic livestock. Takin conservation in Bhutan requires: (1) a commitment to reduce disturbances from domestic livestock through better herding and animal husbandry practices, (2) environmentally friendly road construction, inclusive of wildlife corridors, (3) establishment of satellite offices and regularizing anti-poaching patrol systems, (4) development of education programs to enlist support for Takin conservation, and (5) encouragement of more research on the ecology and management needs of the species.

The increasing kangaroo occurrence in expanding peri-urban areas can be problematic when kangaroos become aggressive towards people and present a collision risk to motor vehicles. An improved understanding on kangaroo spatial and temporal activity patterns in the peri-urban environment is essential to manage kangaroo–human conflict. In this study, we used GPS telemetry to determine activity patterns of male Eastern Grey Kangaroos (Macropus giganteus) in a peri-urban community on the north-coast of New South Wales, Australia. Two types of GPS devices were employed; collars and cheaper alternative glue-on units. Kangaroos moved on average 2.39 km a day, with an average movement rate of 1.89 m/min, which was greatest at dawn. The GPS glue-on devices had short deployment lengths of one to 12 days. Despite limitations in attachment time, the glue-on devices were viable in obtaining daily spatial and temporal activity data. Our results aid towards alleviating conflict with kangaroos by providing new insights into kangaroo movements and activity within a peri-urban environment and introduces a potential cheap GPS alternative for obtaining this data relative to more expensive collars.

Anthropogenic activities and associated global climate change are threatening the biodiversity in the Himalayas against a backdrop of poor knowledge of the region's threatened species. The red panda (Ailurus fulgens) is a threatened mammal confined to the eastern Himalayas, and because of Bhutan's central location in the distributional range of red pandas, its forests are integral to the long-term viability of wild populations. Detailed habitat requirements of the red panda are largely speculative, and there is virtually no ecological information available on this species in Bhutan. Between 2007 and 2009, we established 615 presence/absence plots in a systematic sampling of resident habitat types within Jigme Dorji and Thrumshingla National Parks, Bhutan, to investigate broad and fine-scale red panda habitat associations. Additional locality records of red pandas were obtained from interviewing 664 park residents. Red pandas were generally confined to cool broadleaf and conifer forests from 2,110-4,389 m above sea level (asl), with the majority of records between 2,400-3,700 m asl on south and east-facing slopes. At a finer scale, multivariate analysis revealed that red pandas were strongly associated with old growth Bhutan Fir (Abies densa) forest dominated by a dense cover of Yushania and Arundanaria bamboo with a high density of fallen logs and tree stumps at ground level; a high density of trees, dead snags, and rhododendron shrubs in the mid-storey; and locations that were close to water. Because Bhutan's temperate forests that encompass prime red panda habitat are also integral to human subsistence and socio-economic development, there exists an inadvertent conflict between the needs of people and red pandas. As such, careful sustainable management of Bhutan's temperate forests is necessary if a balance is to be met between the socioeconomic needs of people and the conservation goals for red pandas.

Two α-isoforms of the Na + ,K + -ATPase (α 1 and α 2 ) are expressed in the cardiovascular system, and it is unclear which isoform is the preferential regulator of contractility. Mice heterozygous for the familial hemiplegic migraine type 2 (FHM2) associated mutation in the α 2 -isoform (G301R; α 2 +/G301R mice) have decreased expression of cardiac α 2 -isoform but elevated expression of the α 1 -isoform. We aimed to investigate the contribution of the α 2 -isoform function to the cardiac phenotype of α 2 +/G301R hearts. We hypothesized that α 2 +/G301R hearts exhibit greater contractility due to reduced expression of cardiac α 2 -isoform. Variables for contractility and relaxation of isolated hearts were assessed in the Langendorff system without and in the presence of ouabain (1 µM). Atrial pacing was performed to investigate rate-dependent changes. The α 2 +/G301R hearts displayed greater contractility than WT hearts during sinus rhythm, which was rate-dependent. The inotropic effect of ouabain was more augmented in α 2 +/G301R hearts than in WT hearts during sinus rhythm and atrial pacing. In conclusion, cardiac contractility was greater in α 2 +/G301R hearts than in WT hearts under resting conditions. The inotropic effect of ouabain was rate-independent and enhanced in α 2 +/G301R hearts, which was associated with increased systolic work.

Heterozygous mice (α 2 +/G301R mice) for the migraine-associated mutation (G301R) in the Na + ,K + -ATPase α 2 -isoform have decreased expression of cardiovascular α 2 -isoform. The α 2 +/G301R mice exhibit a pro-contractile vascular phenotype associated with decreased left ventricular ejection fraction. However, the integrated functional cardiovascular consequences of this phenotype remain to be addressed in vivo. We hypothesized that the vascular response to α 2 -isoform-specific inhibition of the Na + ,K + -ATPase by ouabain is augmented in α 2 +/G301R mice leading to reduced cardiac efficiency. Thus, we aimed to assess the functional contribution of the α 2 -isoform to in vivo cardiovascular function of wild-type (WT) and α 2 +/G301R mice. Blood pressure, stroke volume, heart rate, total peripheral resistance, arterial dP/dt, and systolic time intervals were assessed in anesthetized WT and α 2 +/G301R mice. To address rate-dependent cardiac changes, cardiovascular variables were compared before and after intraperitoneal injection of ouabain (1.5 mg/kg) or vehicle during atrial pacing. The α 2 +/G301R mice showed an enhanced ouabain-induced increase in total peripheral resistance associated with reduced efficiency of systolic development compared to WT. When the hearts were paced, ouabain reduced stroke volume in α 2 +/G301R mice. In conclusion, the ouabain-induced vascular response was augmented in α 2 +/G301R mice with consequent suppression of cardiac function.

Objective: Investigating the cardiovascular system is challenging due to its complex regulation by humoral and neuronal factors. Despite this complexity, many existing research methods are limited to the assessment of a few parameters leading to an incomplete characterization of cardiovascular function. Thus, we aim to establish a murine in vivo model for integrated assessment of the cardiovascular system under conditions of controlled heart rate. Utilizing this model, we assessed blood pressure, cardiac output, stroke volume, total peripheral resistance, and electrocardiogram (ECG)., Hypothesis: We hypothesize that (i) our in vivo model can be utilized to investigate cardiac and vascular responses to pharmacological intervention with the α 1 -agonist phenylephrine, and (ii) we can study cardiovascular function during artificial pacing of the heart, modulating cardiac function without a direct vascular effect., Methods: We included 12 mice that were randomly assigned to either vehicle or phenylephrine intervention through intraperitoneal administration. Mice were anesthetized with isoflurane and intubated endotracheally for mechanical ventilation. We measured blood pressure via a solid-state catheter in the aortic arch, blood flow via a probe on the ascending aorta, and ECG from needle electrodes on the extremities. Right atrium was electrically paced at a frequency ranging from 10 to 11.3 Hz before and after either vehicle or phenylephrine administration., Results: Phenylephrine significantly increased blood pressure, stroke volume, and total peripheral resistance compared to the vehicle group. Moreover, heart rate was significantly decreased following phenylephrine administration. Pacing significantly decreased stroke volume and cardiac output both prior to and after drug administration. However, phenylephrine-induced changes in blood pressure and total peripheral resistance were maintained with increasing pacing frequencies compared to the vehicle group. Total peripheral resistance was not significantly altered with increasing pacing frequencies suggesting that the effect of phenylephrine is primarily of vascular origin., Conclusion: In conclusion, this in vivo murine model is capable of distinguishing between changes in peripheral vascular and cardiac functions. This study underlines the primary effect of phenylephrine on vascular function with secondary changes to cardiac function. Hence, this in vivo model is useful for the integrated assessment of the cardiovascular system., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rajanathan, Pedersen, Thomsen, Botker and Matchkov.)

Aims: Acute migraine attack in familial hemiplegic migraine type 2 (FHM2) patients is characterized by sequential hypo- and hyperperfusion. FHM2 is associated with mutations in the Na, K-ATPase α2 isoform. Heterozygous mice bearing one of these mutations (α2+/G301R mice) were shown to have elevated cerebrovascular tone and, thus, hypoperfusion that might lead to elevated concentrations of local metabolites. We hypothesize that these α2+/G301R mice also have increased cerebrovascular hyperaemic responses to these local metabolites leading to hyperperfusion in the affected part of the brain., Methods and Results: Neurovascular coupling was compared in α2+/G301R and matching wild-type (WT) mice using Laser Speckle Contrast Imaging. In brain slices, parenchymal arteriole diameter and intracellular calcium changes in neuronal tissue, astrocytic endfeet, and smooth muscle cells in response to neuronal excitation were assessed. Wall tension and smooth muscle membrane potential were measured in isolated middle cerebral arteries. Quantitative polymerase chain reaction, western blot, and immunohistochemistry were used to assess the molecular background underlying the functional changes. Whisker stimulation induced larger increase in blood perfusion, i.e. hyperaemic response, of the somatosensory cortex of α2+/G301R than WT mice. Neuronal excitation was associated with larger parenchymal arteriole dilation in brain slices from α2+/G301R than WT mice. These hyperaemic responses in vivo and ex vivo were inhibited by BaCl2, suggesting involvement of inward-rectifying K+ channels (Kir). Relaxation to elevated bath K+ was larger in arteries from α2+/G301R compared to WT mice. This difference was endothelium-dependent. Endothelial Kir2.1 channel expression was higher in arteries from α2+/G301R mice. No sex difference in functional responses and Kir2.1 expression was found., Conclusion: This study suggests that an abnormally high cerebrovascular hyperaemic response in α2+/G301R mice is a result of increased endothelial Kir2.1 channel expression. This may be initiated by vasospasm-induced accumulation of local metabolites and underlie the hyperperfusion seen in FHM2 patients during migraine attack., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.)

Familial hemiplegic migraine type 2 (FHM2) is associated with inherited point-mutations in the Na,K-ATPase α2 isoform, including G301R mutation. We hypothesized that this mutation affects specific aspects of vascular function, and thus compared cerebral and systemic arteries from heterozygote mice bearing the G301R mutation (Atp1a2 +/-G301R ) with wild type (WT). Middle cerebral (MCA) and mesenteric small artery (MSA) function was compared in an isometric myograph. Cerebral blood flow was assessed with Laser speckle analysis. Intracellular Ca 2+ and membrane potential were measured simultaneously. Protein expression was semi-quantified by immunohistochemistry. Protein phosphorylation was analysed by Western blot. MSA from Atp1a2 +/-G301R and WT showed similar contractile responses. The Atp1a2 +/-G301R MCA constricted stronger to U46619, endothelin and potassium compared to WT. This was associated with an increased depolarization, although the Ca 2+ change was smaller than in WT. The enhanced constriction of Atp1a2 +/-G301R MCA was associated with increased cSrc activation, stronger sensitization to [Ca 2+ ] i and increased MYPT1 phosphorylation. These differences were abolished by cSrc inhibition. Atp1a2 +/-G301R mice had reduced resting blood flow through MCA in comparison with WT mice . FHM2-associated mutation leads to elevated contractility of MCA due to sensitization of the contractile machinery to Ca 2+ , which is mediated via Na,K-ATPase/Src-kinase/MYPT1 signalling.

New Findings: What is the topic of this review? In this review, we consider the role of the Na + ,K + -ATPase in cerebrovascular function and how it might be changed in familial hemiplegic migraine type 2 (FHM2). The primary focus is involvement of the Na + ,K + -ATPase isoforms in regulation of cerebrovascular tone. What advances does it highlight? In this review, we discuss three overall distinct mechanisms whereby the Na + ,K + -ATPase might be capable of regulating cerebrovascular tone. Furthermore, we discuss how changes in the Na + ,K + -ATPase in cerebral arteries might affect brain perfusion and thereby be involved in the pathology of FHM2., Abstract: Familial hemiplegic migraine type 2 (FHM2) has been characterized by biphasic changes in cerebral blood flow during a migraine attack, with initial hypoperfusion followed by abnormal hyperperfusion of the affected hemisphere. We suggested that FHM2-associated loss-of-function mutation(s) in the Na + ,K + -ATPase α2 isoform might be responsible for these biphasic changes in several ways. We found that reduced expression of the α2 isoform leads to sensitization of the contractile machinery to [Ca 2+ ] i via Src kinase-dependent signal transduction. This change in sensitivity might be the underlying mechanism for both abnormally potentiated vasoconstriction and exaggerated vasorelaxation. Moreover, the functional significance of the Na + ,K + -ATPase α2 isoform in astrocytes provides for the possibility of elevated extracellular potassium signalling from astrocytic endfeet to the vascular wall in neurovascular coupling., (© 2019 The Authors. Experimental Physiology © 2019 The Physiological Society.)

To augment mammal conservation in the Eastern Himalayan region, we assessed the resident 255 terrestrial mammal species and identified the 50 most threatened species based on conservation status, endemism, range size, and evolutionary distinctiveness. By using the spatial analysis package letsR and the complementarity core-area method in the conservation planning software Zonation, we assessed the current efficacy of their protection and identified priority conservation areas by comparing protected areas (PAs), land cover, and global ecoregion 2017 maps at a 100 × 100 m spatial scale. The 50 species that were most threatened, geographically restricted, and evolutionarily distinct faced a greater extinction risk than globally nonthreatened and wide-ranging species and species with several close relatives. Small, medium-sized, and data-deficient species faced extinction from inadequate protection in PAs relative to wide-ranging charismatic species. There was a mismatch between current PA distribution and priority areas for conservation of the 50 most endangered species. To protect these species, the skewed regional PA distribution would require expansion. Where possible, new PAs and transboundary reserves in the 35 priority areas we identified should be established. There are adequate remaining natural areas in which to expand current Eastern Himalayan PAs. Consolidation and expansion of PAs in the EH requires strengthening national and regional transboundary collaboration, formulating comprehensive regional land-use plans, diversifying conservation funding, and enhancing information sharing through a consolidated regional database., (© 2018 Society for Conservation Biology.)

The increasing kangaroo occurrence in expanding peri-urban areas can be problematic when kangaroos become aggressive towards people and present a collision risk to motor vehicles. An improved understanding on kangaroo spatial and temporal activity patterns in the peri-urban environment is essential to manage kangaroo⁻human conflict. In this study, we used GPS telemetry to determine activity patterns of male Eastern Grey Kangaroos ( Macropus giganteus ) in a peri-urban community on the north-coast of New South Wales, Australia. Two types of GPS devices were employed; collars and cheaper alternative glue-on units. Kangaroos moved on average 2.39 km a day, with an average movement rate of 1.89 m/min, which was greatest at dawn. The GPS glue-on devices had short deployment lengths of one to 12 days. Despite limitations in attachment time, the glue-on devices were viable in obtaining daily spatial and temporal activity data. Our results aid towards alleviating conflict with kangaroos by providing new insights into kangaroo movements and activity within a peri-urban environment and introduces a potential cheap GPS alternative for obtaining this data relative to more expensive collars.

Anthropogenic activities and associated global climate change are threatening the biodiversity in the Himalayas against a backdrop of poor knowledge of the region's threatened species. The red panda (Ailurus fulgens) is a threatened mammal confined to the eastern Himalayas, and because of Bhutan's central location in the distributional range of red pandas, its forests are integral to the long-term viability of wild populations. Detailed habitat requirements of the red panda are largely speculative, and there is virtually no ecological information available on this species in Bhutan. Between 2007 and 2009, we established 615 presence/absence plots in a systematic sampling of resident habitat types within Jigme Dorji and Thrumshingla National Parks, Bhutan, to investigate broad and fine-scale red panda habitat associations. Additional locality records of red pandas were obtained from interviewing 664 park residents. Red pandas were generally confined to cool broadleaf and conifer forests from 2,110-4,389 m above sea level (asl), with the majority of records between 2,400-3,700 m asl on south and east-facing slopes. At a finer scale, multivariate analysis revealed that red pandas were strongly associated with old growth Bhutan Fir (Abies densa) forest dominated by a dense cover of Yushania and Arundanaria bamboo with a high density of fallen logs and tree stumps at ground level; a high density of trees, dead snags, and rhododendron shrubs in the mid-storey; and locations that were close to water. Because Bhutan's temperate forests that encompass prime red panda habitat are also integral to human subsistence and socio-economic development, there exists an inadvertent conflict between the needs of people and red pandas. As such, careful sustainable management of Bhutan's temperate forests is necessary if a balance is to be met between the socioeconomic needs of people and the conservation goals for red pandas.