Occupancy,density, and activity patterns of a Critically Endangered leopard population on the Kawthoolei-Thailand border

Large carnivores have been largely extirpated from Southeast Asia due to deforestation, habitat fragmentation, and poaching. Estimating the density of endangered carnivore populations, and identifying relationships between species occupancy and both environmental and anthropogenic factors, is essential for effective conservation planning. Recently, the IUCN conservation status of the Indochinese leopard (Panthera pardus delacouri) was upgraded to “Critically Endangered.” We surveyed Kweekoh Wildlife Sanctuary in Kawthoolei, an area administered by the Karen ethnic group in eastern Myanmar, to quantify (1) leopard population density using spatially explicit mark-resight (SMR) models, (2) leopard occupancy as influenced by important ecological variables, and (3) potential differences in activity between melanistic and spotted leopard morphs. Leopard density was estimated to be 1.39 ± SE 0.22/100 km². Leopard occupancy (ψ = 0.43; 95% credible interval: 0.26–0.67) increased further from roads, at relatively higher elevations, and in areas with higher relative abundance of wild boar. Leopard activity was cathemeral, with higher activity during night hours, and significant overlap (Δ = 0.84; 95% confidence interval: 0.71–0.96) between melanistic and spotted morphs. However, melanistic leopards were more active during twilight hours than spotted individuals whose activity did not significantly vary throughout the day. Indochinese leopard density estimates in Kweekoh were among the lowest reported from Southeast Asia. Leopard occupancy was highest in the sanctuary's core areas, suggesting the presence of negative anthropogenic impacts along the sanctuary borders. We suggest our low density estimates warrant immediate and decisive conservation action, including better protection for leopards, their habitat, and their prey.     

A select panel of polymorphic microsatellite loci for individual identification of snow leopards (Panthera uncia)

Snow leopards (Panthera uncia) are elusive endangered carnivores found in remote mountain regions of Central Asia. New methods for identifying and counting snow leopards are needed for conservation and management efforts. To develop molecular genetic tools for individual identification of hair and faecal samples, we screened 50 microsatellite loci developed for the domestic cat (Felis catus) in 19 captive snow leopards. Forty‐eight loci were polymorphic with numbers of alleles per locus ranging from two to 11. The probability of observing matching genotypes for unrelated individuals (2.1 × 10^?11) and siblings (7.5 × 10^?5) using the 10 most polymorphic loci was low, suggesting that this panel would easily discriminate among individuals in the wild.     

Molecular evidence for species-level distinctions in clouded leopards

Among the 37 living species of Felidae, the clouded leopard (Neofelis nebulosa) is generally classified as a monotypic genus basal to the Panthera lineage of great cats. This secretive, mid-sized (16-23 kg) carnivore, now severely endangered, is traditionally subdivided into four southeast Asian subspecies (Figure 1A). We used molecular genetic methods to re-evaluate subspecies partitions and to quantify patterns of population genetic variation among 109 clouded leopards of known geographic origin (Figure 1A, Tables S1 ans S2 in the Supplemental Data available online). We found strong phylogeographic monophyly and large genetic distances between N. n. nebulosa (mainland) and N. n. diardi (Borneo; n = 3 individuals) with mtDNA (771 bp), nuclear DNA (3100 bp), and 51 microsatellite loci. Thirty-six fixed mitochondrial and nuclear nucleotide differences and 20 microsatellite loci with nonoverlapping allele-size ranges distinguished N. n. nebulosa from N. n. diardi. Along with fixed subspecies-specific chromosomal differences, this degree of differentiation is equivalent to, or greater than, comparable measures among five recognized Panthera species (lion, tiger, leopard, jaguar, and snow leopard). These distinctions increase the urgency of clouded leopard conservation efforts, and if affirmed by morphological analysis and wider sampling of N. n. diardi in Borneo and Sumatra, would support reclassification of N. n. diardi as a new species (Neofelis diardi).     

Twenty-two polymorphic microsatellite loci aimed at detecting illegal trade in the Cape parrot, Poicephalus robustus (Psittacidae, AVES)

Twenty-two polymorphic microsatellite loci were characterized in the Cape parrot, Poicephalus robustus. Nineteen loci were newly isolated from two Cape parrot genomic libraries, and three loci isolated from other parrot species. Loci were characterized in 40 unrelated captive Cape parrots held by aviculturalists. The loci displayed between two and 24 alleles, with the observed heterozygosities ranging between 0.10 and 0.94. This locus set is suitable for identifying clarifying parentage (parentage exclusion probabilities of P_E1 = 0.0004 and P_E2 = 0.000001). Candidate parents for any Cape parrot individual can now be genotyped to distinguish between individuals, which are truly captive bred and those suspected of being wild-caught birds. Cross-species analysis found up to 31 loci to be polymorphic across 24 additional parrot species tested.     

Population dynamics of pedigree leopards,Panthera pardus ssp,in captivity

The demographic history of 4 races or subspecies of leopard, Panthera pardus, was reviewed from international studbook records dating back to 1953. The Chinese leopard has been the most common pedigree race maintained in captivity, a factor linked to the length of time (29 years) this subspecies has been in captivity. The relative youth of the wild-born founders also helped them to adjust to captivity as well as live long reproductive lives. Today, however, this race is suffering from the ill effects of inbreeding due to the small founder size. This condition appears to be correctable now that additional specimens have been located. Persian leopards have a larger founder size than the former race, but some of their ancestors were older animals at the time of acquisition. Because of this, their potential fecundity was probably depressed from psychological problems related to adjustment and a shorter life span in captivity. Two founding females experienced pelvic deformities while young, and few of their cubs survived because they all had to be delivered via caesarian section. This procedure also shortened the reproductive life of the females involved because the owning zoos refrained from breeding the animals in the leopards' later years. Captive leopards appear to live longer than their wild counterparts, although precise data on wild populations is not available. In captivity many reach 12–15 years old, and exceptional individuals of several races have lived 20 years. Most captive-born leopards begin breeding when they are 3 years old and continue until they are 8–10 years old. Reproduction in females usually ceases at 12–14 years, although males have a longer reproductive life, with several successfully breeding when 19–20 years old.     

Geographical variation in and evolutionary history of the Sunda clouded leopard (Neofelis diardi) (Mammalia: Carnivora: Felidae) with the description of a new subspecies from Borneo

Recent morphological and molecular studies led to the recognition of two extant species of clouded leopards; Neofelis nebulosa from mainland southeast Asia and Neofelis diardi from the Sunda Islands of Borneo and Sumatra, including the Batu Islands. In addition to these new species-level distinctions, preliminary molecular data suggested a genetic substructure that separates Bornean and Sumatran clouded leopards, indicating the possibility of two subspecies of N. diardi. This suggestion was based on an analysis of only three Sumatran and seven Bornean individuals. Accordingly, in this study we re-evaluated this proposed subspecies differentiation using additional molecular (mainly historical) samples of eight Bornean and 13 Sumatran clouded leopards; a craniometric analysis of 28 specimens; and examination of pelage morphology of 20 museum specimens and of photographs of 12 wild camera-trapped animals. Molecular (mtDNA and microsatellite loci), craniomandibular and dental analyses strongly support the differentiation of Bornean and Sumatran clouded leopards, but pelage characteristics fail to separate them completely, most probably owing to small sample sizes, but it may also reflect habitat similarities between the two islands and their recent divergence. However, some provisional discriminating pelage characters are presented that need further testing. According to our estimates both populations diverged from each other during the Middle to Late Pleistocene (between 400 and 120 kyr). We present a discussion on the evolutionary history of Neofelis diardi sspp. on the Sunda Shelf, a revised taxonomy for the Sunda clouded leopard, N. diardi, and formally describe the Bornean subspecies, Neofelis diardi borneensis, including the designation of a holotype (BM.3.4.9.2 from Baram, Sarawak) in accordance with the rules of the International Code of Zoological Nomenclature.     

Genetic diversity and species identification in the endangered white abalone (<Emphasis Type="Italic">Haliotis sorenseni</Emphasis>)

In 2001, the white abalone Haliotis sorenseni became the first marine invertebrate in United States waters to receive federal protection as an endangered species. Prior to the endangered species listing, 20 abalone were collected as potential broodstock for a captive rearing program. Using DNA from these animals, we have developed genetic markers, including five nuclear microsatellite loci and partial sequences of one nuclear (VERL) and two mitochondrial (COI and CytB) genes, to assess genetic variability in the species, aid in species identification, and potentially track the success of future outplanting of captive-reared animals. All five microsatellite loci were polymorphic and followed expectations of simple Mendelian inheritance in laboratory crosses. Each of the wild-caught adult abalone exhibited a unique composite microsatellite genotype, suggesting that significant genetic variation remains in natural populations. A combination of nuclear and mitochondrial gene sequencing demonstrated that one of the original wild-caught animals was, in fact, not a white abalone, but H. kamtschatkana (possibly subspecies assimilis). Similarly, another animal of uncertain identity accidentally collected by dredging was also shown to be H. kamtschatkana. Inclusion of these two animals as broodstock could have resulted in unintentional hybridizations detrimental to the white abalone recovery program. Molecular genetic identifications will be useful both in preventing broodstock contamination and as markers for future restocking operations.     

Examining genetic diversity and identifying polymorphic microsatellite markers for noninvasive genetic sampling of the Indochinese leopard (Panthera pardus delacouri)

Indochinese leopard (Panthera pardus delacouri) is one of the least-known leopard subspecies and occurs in mainland South-east Asia. Given the elusive nature, low density, and large movement ranges of leopard, noninvasive genetic sampling may be an effective approach for studying this subspecies. We examined the potential for 18 microsatellite loci to allow reliable identification of noninvasively collected leopard samples based on feces collected in eastern Cambodia. The expected heterozygosity calculated from 5 to 7 leopard individuals ranged from 0.49 to 0.91 with an average of 0.73, showing a high level of genetic diversity. Only the five most informative loci decreased the probability of identity for siblings (P_ID-sib) to less than 0.01, which would be sufficient for abundance estimation. A further increase of the number of loci up to 8–10 decreased the P_ID-sib to the level of 0.001–0.0001, which may be useful in cases of parentage analysis or forensic analysis. The microsatellite markers shown in the present study achieved a sufficient level of accuracy for individual identification, and thus, will be useful for examining abundance, genetic structure, or relatedness of the Indochinese leopard in future.     

Forest cover and level of protection influence the island-wide distribution of an apex carnivore and umbrella species,the Sri Lankan leopard (<Emphasis Type="Italic">Panthera pardus kotiya</Emphasis>)

Apex predators fulfil potentially vital ecological roles. Typically wide-ranging and charismatic, they can also be useful surrogates for biodiversity preservation, making their targeted conservation imperative. The Sri Lankan leopard (Panthera pardus kotiya), an endangered, endemic sub-species, is the island’s apex predator. Of potential keystone importance, this carnivore also fulfills “umbrella” and “flagship” criterion and is of high ecological and existence value. Apex predator conservation requires identifying factors underlying distribution, so we used multi-scale maximum entropy modelling with sampling bias correction to investigate a broad suite of relevant ecological, climatic and anthropogenic factors in order to identify potentially suitable leopard habitat. Presence locations were determined from 15 years of surveys, observations and verified reports. The best bias correction procedure and scale were uncertain, so we employed a novel method of using information from all models across analyses to determine top models and identify influential variables. Leopard presence was most strongly linked to the landscape proportion encompassed by Protected Areas strictly limiting human presence, with more porous Protected Areas less influential. All three forest composition and configuration metrics investigated (area weighted mean patch size, patch density and forest connectivity) were influential, with increased patch size and higher connectivity predicting better habitat suitability for leopards. Habitat suitability was also better where cropland extent and urban patch size were small. In summary, ground-level protection and natural forest extent and connectivity are of profound importance to Sri Lankan leopard distribution and are key factors in ensuring the ecological integrity of the island’s faunal assemblages.     

Distribution and human‐caused mortality of Persian leopards Panthera pardus saxicolor in Iran,based on unpublished data and Farsi gray literature

Gray literature and data from unpublished sources can provide important scientific information that has not been published scientifically. The Persian leopard (hereafter leopard) Panthera pardus saxicolor is classed as endangered on the Red List of the International Union for Conservation of Nature and also is one of the least‐studied subspecies of leopard. It occurs in the Caucasus and Central and Southwest Asia. Iran contains more than 75% of the leopard's extant range, and the leopard population in this country serves as a source for neighboring countries. In this study, we determined the distribution and human‐caused mortality of leopards in Iran, by reviewing unpublished data and Farsi gray literature (which includes government reports) between 1 January 2010 and 30 December 2018. We created the most recent distribution map of the leopard in Iran. Our data display that human‐caused mortality of leopard in Iran mostly includes poaching and intentional poisoning, and roadkill.     

Population genetics and conservation of the threatened southeastern beach mouse (Peromyscus polionotus niveiventris): subspecies and evolutionary units

We investigated genetic diversity within the southeastern beach mouse (SEBM-Peromyscus polionotus niveiventris) and also tested the hypothesis that the subspecies recognition of P.p. niveiventris, based on size and color differences, is congruent with this taxon representing a discrete evolutionary lineage. We used ten polymorphic microsatellite loci and mitochondrial cytochrome-b gene DNA sequences to investigate genetic diversity and population structure within the SEBM, and to determine the level of divergence between the SEBM and the nearest known inland subspecies of the oldfield mouse (Peromyscus polionotus rhoadsi). Moderate genetic distances were observed between the SEBM and the inland oldfield mouse based on microsatellite data, with F _ST values ranging from 0.11 to 0.22 between these taxa. Additionally, mitochondrial DNA haplotypes of the SEBM formed a distinct monophyletic group relative to haplotypes sampled from P. p. rhoadsi. Based on previous estimates of rates of mitochondrial DNA evolution in rodents, we inferred that Pleistocene sea-level fluctuations are likely responsible for the historical isolation of the SEBM lineage from mainland P. polionotus. Our data demonstrate the genetic distinctiveness of the SEBM, justifying the current subspecies designation for the SEBM and its continued protection under the United States Endangered Species Act. We classify the Cape Canaveral and Smyrna Dunes Park populations of SEBM as a single evolutionary significant unit. The two known extant allopatric populations of the SEBM showed some differentiation in microsatellite frequencies and were moderately reciprocally distinguishable based on assignment to distinct genetic clusters by a Bayesian admixture procedure. These results justify the classification of these two extant SEBM populations as distinct management units that should be independent targets of management and conservation attention.     

Evidence for African origins of founders of the asiatic lion species survival plan

The Asiatic lion (Panthera leo persica) exists in the wild as a single relict population of approximately 250 individuals in the protected Gir Forest Sanctuary in western India. In 1981, a species survival plan (SSP) for the Asiatic lion was established by the American Association of Zoological Parks and Aquariums to manage the 200 + descendants of Asiatic lions in captivity in western zoological facilities. This captive population was derived from seven founders. In order to compare the genetic structure of the Gir Forest population with that of the captive SSP population, a genetic survey of 46 electrophoretic allozyme systems resolved from extracts of lion blood was undertaken by using 29 SSP Asiatic lions and 28 wild-caught or captive-bred lions maintained at the Sakkarbaug Zoo in India but originally derived from the Gir Forest. The Gir lion population was found to be genetically monomorphic at each of 46 allozyme loci. This was in contrast to several African lion (Panthera leo leo) populations, which show moderate levels of allozyme variation at the same loci. The SSP lion population was polymorphic at three allozyme loci (IDHI, TF, and PTI) for alleles that were previously found only in African lion populations. Pedigree analysis of the genetic transmission of these three biochemical loci demonstrated that two of the five primary founder animals of the SSP Asiatic lion population (a breeding pair originally imported from the Trivandrum Zoo in southern India) were descendants of the African subspecies. Three other founder animals were pure Asian. A retrospective SSP pedigree analysis of two morphologic characters (prominent abdominal fold and pairing of infraorbital foramen) that are partially diagnostic for persica vs leo was consistent with this conclusion as well. The implications for the management of small captive populations of threatened species and of the Asiatic lion SSP population are discussed.     

Phylogenetics, genome diversity and origin of modern leopard, Panthera pardus

Leopards, Panthera pardus, are widely distributed across southern Asia and sub-Saharan Africa. The extent and phylogeographic patterns of molecular genetic diversity were addressed in a survey of 77 leopards from known geographical locales representing 13 of the 27 classical trinomial subspecies. Phylogenetic analysis of mitochondrial DNA sequences (727 bp of NADH5 and control region) and 25 polymorphic microsatellite loci revealed abundant diversity that could be partitioned into a minimum of nine discrete populations, tentatively named here as revised subspecies: P. pardus pardus, P. p. nimr, P. p. saxicolor, P. p. fusca, P. p. kotiya, P. p. delacouri, P. p. japonensis, P. p. orientalis and P. p. melas. However, because of limited sampling of African populations, this may be an underestimate of modern phylogeographic population structure. Combined phylogeographic and population diversity estimates support an origin for modern leopard lineages 470,000-825,000 years ago in Africa followed by their migration into and across Asia more recently (170,000-300,000 years ago). Recent demographic reductions likely have led to genetic impoverishment in P. p. orientalis and in the island subspecies P. p. kotiya.     

Directional selection in the evolution of elongated upper canines in clouded leopards and sabre‐toothed cats

Extremely developed or specialized traits such as the elongated upper canines of extinct sabre‐toothed cats are often not analogous to those of any extant species, which limits our understanding of their evolutionary cause. However, an extant species may have undergone directional selection for a similar extreme phenotype. Among living felids, the clouded leopard, Neofelis nebulosa, has exceptionally long upper canines for its body size. We hypothesized that directional selection generated the elongated upper canines of clouded leopards in a manner similar to the process in extinct sabre‐toothed cats. To test this, we developed an approach that compared the effect of directional selection among lineages in a phylogeny using a simulation of trait evolution and approximate Bayesian computation. This approach was applied to analyse the evolution of upper canine length in the Felidae phylogeny. Our analyses consistently showed directional selection favouring longer upper canines in the clouded leopard lineage and a lineage leading to the sabre‐toothed cat with the longest upper canines, Smilodon. Most of our analyses detected an effect of directional selection for longer upper canines in the lineage leading to another sabre‐toothed cat, Homotherium, although this selection may have occurred exclusively in the primitive species. In all the analyses, the clouded leopard and Smilodon lineages showed comparable directional selection. This implies that clouded leopards share a selection advantage with sabre‐toothed cats in having elongated upper canines.     

Carnivores from Los Rincones,a leopard den in the highest mountain of the Iberian range (Moncayo,Zaragoza, Spain)

Carnivores are a common element in Pleistocene fossil assemblages. However, they are not so abundant in terms of the numbers of remains. Here, we present a palaeontological study of the carnivores from the Late Pleistocene (MIS 3) of Los Rincones (Spain), one of the few deposits accumulated by leopards. One hundred and ten leopard remains have been recovered. This carnivore is not the only inhabitant of the cave, and 175 remains belonging to Ursus arctos have also been recovered, making it one of the sites with the greatest number of brown bear remains in the Iberian Peninsula. The large number of leopard remains has allowed us to make a detailed morphological and biometrical study that has enabled us to classify the remains within the subspecies Panthera pardus spelaea. The European Ice Age leopard inhabited Europe during Upper Pleistocene and it presents some similarities with Panthera uncia. A study of the scarce remains of Canis lupus indicates that this was similar in size to Canis lupus maximus; the scarcity of the remains prevents us from assigning our remains to this subspecies. A study of the brown bear remains indicates that it is similar to other populations in the north of the Iberian Peninsula with this chronology.     

圈养金钱豹行为谱

2005年3月～2006年3月,采取所有事件取样法和目标取样法,对成都动物园圈养的17只金钱豹进行行为观察,在此基础卜制定了行为谱.研究表明:休息、观望、运动是圈养金钱豹的主要行为,占其所有行为的93.1%,休息占其所有行为的54.40%;观望占21.18%;运动占17.53%.     

Evaluation of the role of the national parks for Persian leopard (<Emphasis Type="Italic">Panther pardus saxicolor</Emphasis>, Pocock 1927) habitat conservation (case study: Tandooreh National Park,Iran)

Having knowledge of the habitat requirements of top predators is important for their conservation and also for the stability of wildlife communities since they are keystone species of ecosystems. This study aims to predict suitable habitats for the Persian leopard (Panther pardus saxicolor, Pocock 1927) in Tandooreh National Park, Iran. This species is the largest leopard subspecies in the Middle East and is at high risk of extinction. Presence points for the species have been obtained from field data, literature review, atlas data, and IUCN records. Variables related to human disturbance, terrain, land use/land cover, and the leopards’ prey were used to predict the habitat selection of the Persian leopard by employing binary logistic regression. Our model predicted that the presence of Persian leopards is restricted to the southern, north-western, and western areas of the park which are under severe fragmentation threat. Moreover, leopards avoided areas near human development such as roads. We have also found that the availability of prey such as Capra aegagrus, Ovis vignei, and Ovis orientalis is one of the important parameters to predict habitat suitability for leopards. Our model provides a suitable tool for national park managers to identify biodiversity hotspots since leopards are often associated with high biodiversity.     

Genetic differentiation among local populations of Asian elephant

Electrohoretically detectable enetic variation for 29 kinds of blood protein encoded by 33 loci was analyzed for 78 Asian eletants (Elephas maximus) which were collected from its four local populations: Sri Lanka, Souti India, Thailand and Nepal. Elehants in Sri Lanka are classified into the subspecies E.m. maximus, and those from the other tlree localities into the subspecies E. m. indicus. Six variable loci were detected, and one of them, the tetrazolium oxidase locus, was observed to show a complete allele substitution between the subspecies. Average heterozgosity within local populations were in a range of 0.0152 ? 0.0303. Whereas the Nei's genetic distance among three local populations of the subspecies indicus were 0.0013 ? 0.0031, the distance between the subspecies indicus and maximus were 0.0328 ? 0.0370, indicating that the two subspecies were well differentiated genetically.     

Observations on reproduction,hormones, copulatory behavior,and neonatal mortality in captive Lemur mongoz (Mongoose lemur)

This study examines correlates of reproduction in Lemur mongoz, an endangered prosimian primate with a history of poor captive reproduction. Male testicular growth and regression were quantified via testicular indices (TI). Although TIs could be used to predict male breeding condition, they were not indicators of ability to sire offspring. Serum estradiol (E₂) and progesterone (P) concentrations, along with vaginal cytology, were used to monitor female reproductive cyclicity. E₂ and P profiles showed no abnormalities. An elevation of E₂ with low P, approximately one estrous cycle length (31.4 days) prior to the onset of fertile cycles, was consistent with the phenomenon of ‘pseudoestrus’ observed in other lemur species. The copulatory pattern, consisting of thrusting, multiple intromissions, single or multiple ejaculations, and no copulatory lock, was more complex than expected for an animal with a supposedly monogamous mating system. The preference for natural arboreal substrata during copulation suggests that enclosure furnishings may influence copulatory behavior. A 30% neonatal mortality rate tempered recent success in breeding L. mongoz at the Duke University Primate Center (DUPC). Although infant sex and parity of the mother were not predictive of neonatal mortality, mother's natal status was, with captive-born females having a 67% infant mortality rate, compared with 15% in wild-caught females. Based on these observations, a number of factors, including housing, behavior, and photoperiod, may be relevant to successful captive propagation of L. mongoz. © 1992 Wiley-Liss, Inc.