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Of the sub-species of Holarctic wolf, the Woolly wolf (Canis lupus chanco) is uniquely adapted to atmospheric hypoxia and widely distributed across the Himalaya, Qinghai Tibetan Plateau (QTP) and Mongolia. Taxonomic ambiguity still exists for this sub-species because of complex evolutionary history anduse of limited wild samples across its range in Himalaya. We document for the first time population genetic structure and taxonomic affinity of the wolves across western and eastern Himalayan regions from samples collected from the wild (n = 19) using mitochondrial control region (225bp). We found two haplotypes in our data, one widely distributed in the Himalaya that was shared with QTP and the other confined to Himachal Pradesh and Uttarakhand in the western Himalaya, India. After combining our data withpublished sequences (n = 83), we observed 15 haplotypes. Some of these were shared among different locations from India to QTP and a few were private to geographic locations. A phylogenetic tree indicated that Woolly wolves from India, Nepal, QTP and Mongolia are basal to other wolves with shallow divergence (K2P; 0.000-0.044) and high bootstrap values. Demographic analyses based on mismatch distribution and Bayesian skyline plots (BSP) suggested a stable population over a long time (~million years) with signs of recent declines. Regional dominance of private haplotypes across its distribution range may indicate allopatric divergence. This may be due to differences in habitat characteristics, availability of different wild prey species and differential deglaciation within the range of the Woolly wolf during historic time. Presence of basal and shallow divergence within-clade along with unique ecological requirements and adaptation to hypoxia, the Woolly wolf of Himalaya, QTP, and Mongolian regions may be considered as a distinct an Evolutionary Significant Unit (ESU). Identifying management units (MUs) is needed within its distribution range using harmonized multiple genetic data for effective conservation planning.

The Sundarbans tiger inhabits a unique mangrove habitat and are morphologically distinct from the recognized tiger subspecies in terms of skull morphometrics and body size. Thus, there is an urgent need to assess their ecological and genetic distinctiveness and determine if Sundarbans tigers should be defined and managed as separate conservation unit. We utilized nine microsatellites and 3 kb from four mitochondrial DNA (mtDNA) genes to estimate genetic variability, population structure, demographic parameters and visualize historic and contemporary connectivity among tiger populations from Sundarbans and mainland India. We also evaluated the traits that determine exchangeability or adaptive differences among tiger populations. Data from both markers suggest that Sundarbans tiger is not a separate tiger subspecies and should be regarded as Bengal tiger (P. t. tigris) subspecies. Maximum likelihood phylogenetic analyses of the mtDNA data revealed reciprocal monophyly. Genetic differentiation was found stronger for mtDNA than nuclear DNA. Microsatellite markers indicated low genetic variation in Sundarbans tigers (He= 0.58) as compared to other mainland populations, such as northern and Peninsular (Hebetween 0.67- 0.70). Molecular data supports migration between mainland and Sundarbans populations until very recent times. We attribute this reduction in gene flow to accelerated fragmentation and habitat alteration in the landscape over the past few centuries. Demographic analyses suggest that Sundarbans tigers have diverged recently from peninsular tiger population within last 2000 years. Sundarbans tigers are the most divergent group of Bengal tigers, and ecologically non-exchangeable with other tiger populations, and thus should be managed as a separate \"evolutionarily significant unit\" (ESU) following the adaptive evolutionary conservation (AEC) concept.

Despite massive global conservation strategies, tiger populations continued to decline until recently, mainly due to habitat loss, human-animal conflicts, and poaching. These factors are known to affect the genetic characteristics of tiger populations and decrease local effective population sizes. The Terai Arc Landscape (TAL) at the foothills of the Himalaya is one of the 42 source sites of tigers around the globe. Therefore, information on how landscape features and anthropogenic factors affect the fine-scale spatial genetic structure and variation of tigers in TAL is needed to develop proper management strategies for achieving long-term conservation goals. We document, for the first time, the genetic characteristics of this tiger population by genotyping 71 tiger samples using 13 microsatellite markers from the western region of TAL (WTAL) of 1800 km2. Specifically, we aimed to estimate the genetic variability, population structure, and gene flow. The microsatellite markers indicated that the levels of allelic diversity (MNA = 6.6) and genetic variation (Ho = 0.50, HE = 0.64) were slightly lower than those reported previously in other Bengal tiger populations. We observed moderate gene flow and significant genetic differentiation (FST= 0.060) and identified the presence of cryptic genetic structure using Bayesian and non-Bayesian approaches. There was low and significantly asymmetric migration between the two main subpopulations of the Rajaji Tiger Reserve and the Corbett Tiger Reserve in WTAL. Sibship relationships indicated that the functionality of the corridor between these subpopulations may be retained if the quality of the habitat does not deteriorate. However, we found that gene flow is not adequate in view of changing land use matrices. We discuss the need to maintain connectivity by implementing the measures that have been suggested previously to minimize the level of human disturbance, including relocation of villages and industries, prevention of encroachment, and banning sand and boulder mining in the corridors.

Big cats (Genus: Panthera ) are among the most threatened mammal groups of the world, owing to hunting, habitat loss, and illegal transnational trade. Conservation genetic studies and effective curbs on poaching are important for the conservation of these charismatic apex predators. A limited number of microsatellite markers exists for Panthera species and researchers often cross-amplify domestic cat microsatellites to study these species. We conducted data mining of seven Panthera genome sequences to discover microsatellites for conservation genetic studies of four threatened big cat species. A total of 32 polymorphic microsatellite loci were identified in silico and tested with 152 big cats, and were found polymorphic in most of the tested species. We propose a set of 12 novel microsatellite markers for use in conservation genetics and wildlife forensic investigations of big cat species. Cumulatively, these markers have a high discriminatory power of one in a million for unrelated individuals and one in a thousand for siblings. Similar PCR conditions of these markers increase the prospects of achieving efficient multiplex PCR assays. This study is a pioneering attempt to synthesise genome wide microsatellite markers for big cats.

The western Terai Arc Landscape (wTAL) in Uttarakhand, India, marks the range limit for the Asian elephant in north‐western India. This region has been impacted by land‐use changes and infrastructure expansion for the last seven decades. To evaluate the impact of habitat deterioration on the population structure of elephants in the region, we characterized their genetic diversity and local genetic structure using mitochondrial (D loop) and nuclear DNA (microsatellites; n = 15) markers. We used tissue samples of 114 elephants from five different sub‐populations, collected between 2005 and 2014. The genetic variation was moderate (HO = 0.49–0.55) compared with other Indian elephant populations. Two mtDNA haplotypes were identified without strong spatial patterns across wTAL. Bayesian individual‐based clustering algorithm identified two genetic clusters (K = 2) with high admixture (50% at Q < 0.7) and no spatial adherence. Though K = 1 was not supported by the Bayesian algorithm, multivariate analysis and sibship patterns did not indicate genetic differentiation. The lack of spatial genetic structuring suggests high levels of gene flow, indicating that this population is still panmictic. This suggests that the life history traits of elephants as well as the ecological features of this landscape influence genetic connectivity. However, ongoing land use changes necessitate regular genetic monitoring in wTAL to identify incipient structuring caused by anthropogenic barriers to movement. Nuclear microsatellite and mitochondrial DNA sequences reveal the panmictic characteristic of the Asian elephant population at their north‐western range limit in India. However, the linear landscape is isolated and undergoing land‐use change and linear infrastructure development. Regular genetic monitoring to detect any incipient structuring would help secure the long‐term viability of the population.

Population estimates of tigers (Panthers tigris) are affected by movement patterns of the sexes. We evaluated sex-specific heterogeneity to estimate the tiger population at Ranthambhore Tiger Reserve, western Rajasthan, India. We also compared population attributes obtained using a likelihood-based spatially explicit capture–recapture (SECR) method with traditional approaches that use alternate information on animal space used to estimate an effective sampled area around the traps. We obtained 122 tiger photocaptures of 27 individuals (14 F and 13 M) from 165 camera-trapping sites in a 215-km2area over 3,300 trapnights, for 20 sampling occasions in 2010. The capture probability of males was about 2 times greater than that of female tigers, but overall detection probability was high, with total population estimate of about 30.2 tigers (SE = 3.7). Male and female tiger density estimates/100 km2were 2.8 (SE = 0.8) and 4.1 (SE = 1.1), respectively, and overall density estimates of 6.1 (SE = 1.2) individuals/100 km2were obtained using the SECR method. Non-spatial models using the full mean maximum distance moved (MMDM) to estimate an effective sampled area underestimated the density by 9% compared with the SECR model as a baseline, and 1/2MMDM models overestimated the density by 25% as compared with results from the SECR model. We conclude that to assess the reliability of abundance estimates, heterogeneity in sex-specific capture probabilities should be considered. We recommend using an SECR model because these methods appear to be minimally biased for estimating an effective sampled area and can be applied consistently among studies of various taxa.

The Asian elephant is mostly confined to mountainous ranges and therefore risks population fragmentation if hard protected area (PA) boundaries near steep slopes prevent movement. We tested whether elephant gene flow is (i) controlled by slope and (ii) affected by the interplay between barriers and slope. We used 176 unique genotypes obtained non-invasively from fresh elephant dung to assess individual-by-individual genetic distance across the Western Ghats of India, a biodiversity hotspot. To assess landscape distance, 36 resistance models were produced by transforming a slope raster. Core areas and corridors were calculated from the raster that provided the best correlation between the genetic and distance matrices. The influence of the closure of PAs on gene flow was examined for one region, the Nilgiri Biosphere Reserve. The best resistance raster obtained by transforming the slope occupancy model was better than Euclidean distance for explaining genetic distance, indicating that slope partially controls gene flow. Fencing elephant PAs on hilly terrain reduces core areas and disrupts corridors. Consequently, hard PA boundaries abutting slopes can fragment elephant populations, but this can be ameliorated by protecting the adjacent flatter terrain.

Increasing habitat loss and fragmentation as a result of anthropogenic influences have been implicated as a major cause in declining wildlife populations. To mitigate the anthropogenic impacts, many countries have developed large reserves to protect important habitats. However, for large carnivores, such as the Bengal tiger (Panthera tigris tigris), reserves may not mitigate human-caused mortality. We monitored a tiger population using cameras and intensive searches from 2005 to 2013 to evaluate the factors causing losses of tigers in Ranthambhore Tiger Reserve, western India. The tiger population in Ranthambhore Tiger Reserve is an important source population and genetic pool in the western-most distribution of tigers in India. Fourteen tigers died during the study period: 5 from human-induced caused and 9 from other causes (natural, depredation hunts, and dispersal). We could not determine the fate of an additional 8 tigers. Continuous monitoring of tiger populations is required, to monitor the factors causingmortality and losses of individuals. Our results offer baseline information on tiger losses in the landscape, which can be used to assist future management and conservation strategies for tigers.

Pangolins are the world`s most trafficked mammalian species classified under family Manidae and face severe threat of extinction, largely due to the illicit trade of its parts and products, especially scales, in international markets. Pangolin scales are believed to be used in Traditional Chinese Medicines (TCM) and meat is used as delicacies in restaurants. Of the eight extant species of pangolin, morphological discrimination is easy but the situation becomes precarious once the scales and meat samples are seized and it is difficult to identify species based on morphology in such cases. However, wildlife DNA forensics has played an instrumental role in the identification of species from such type of materials. The present study investigated that three mitochondrial genes (Cyt b, 16S rRNA, and 12S rRNA) clearly showed the variation among seven extant pangolin species (Manis culionensis; possibly extinct), whereas, maximum variation was obtained in cytochrome b when compared to another two mitochondrial genes. The present study revealed that obtained SNPs based on short sequence length (Intervals) within the three mitochondrial genes will be helpful to design the short molecular marker and species-specific probe that is used in wildlife forensic for identifying pangolin species from the degraded sample. We also advocate using more than one molecular marker for species discrimination so as to minimize any false identification of the mammal's species reported in the trade. Furthermore, data generated from the study would help in strengthening the DNA database of Indian pangolin species.

Bengal tigers are highly endangered and knowledge on adaptive genetic variation can be essential for efficient conservation and management. Here we present the first assessment of allelic variation in major histocompatibility complex (MHC) class I and MHC class II DRB genes for wild and captive tigers from India. We amplified, cloned, and sequenced alpha-1 and alpha-2 domain of MHC class I and beta-1 domain of MHC class II DRB genes in 16 tiger specimens of different geographic origin. We detected high variability in peptide-binding sites, presumably resulting from positive selection. Tigers exhibit a low number of MHC DRB alleles, similar to other endangered big cats. Our initial assessment—admittedly with limited geographic coverage and sample size—did not reveal significant differences between captive and wild tigers with regard to MHC variability. In addition, we successfully amplified MHC DRB alleles from scat samples. Our characterization of tiger MHC alleles forms a basis for further in-depth analyses of MHC variability in this illustrative threatened mammal.