Gastro-intestinal parasites of sympatric red panda and livestock in protected areas of Nepal.

BACKGROUND: Disease transmission among humans, domestic animals and wildlife can have profound consequences in human health, wildlife conservation and maintenance of biodiversity. The issue of disease transmission can be particularly important for threatened wildlife species, yet such information remains scarce due to logistic constraints and government regulation on animal handlings. The red panda (Ailurus fulgens) is one of the globally threatened species challenged with habitat fragmentation and human disturbance. In Nepal, livestock grazing is recognised as one of the major threats to the red panda. AIM: We aimed to provide the first empirical data on gastro-intestinal parasites for sympatric livestock and red panda from two geographically isolated regions in Nepal. METHODS: In this study, we systematically sampled, and examined the faecal of livestock and red panda in two separate protected areas to provide the first empirical data on their gastro-intestinal parasite, including the prevalence, parasite richness and load. RESULTS: We documented 11 parasite taxa (7 nematodes, 2 cestodes, 1 trematode and 1 coccidian), of which 8 are shared by both livestock and red panda. Furthermore, parasite prevalence, parasite load and parasite richness were generally higher in the livestock than the red panda. CONCLUSION: The data provided from this systematic survey on parasites of sympatric livestock and red panda in wild raises the concern about the potential role of livestock mediating disease dynamics in the red panda. Our study suggests that cross-transmission of parasites between livestock and red panda are likely, and the livestock may be a competent agent bringing disease to both red panda and human. Therefore, managing human-livestock-wildlife contact to reduce disease risk to all groups should be a key component in conservation planning of protected areas.

Potential effects of GPS collars on the behaviour of two red pandas (Ailurus fulgens) in Rotterdam Zoo.

GPS collars are frequently used to study the (behavioural) ecology of species. However, such collars can cause behavioural changes and can have negative physiological effects on the individuals wearing them. A pilot study to obtain data on behavioural and physiological effects of GPS collars on the target species would therefore be recommended, especially when it concerns rare or endangered species. The red panda (Ailurus fulgens) is a small carnivore endemic to the mountains of Central Asia that is currently classified as endangered. There is a lack in knowledge on the species ecology which could be enhanced by a study using GPS-technology. As a pilot study, the two adult red pandas in Rotterdam Zoo were observed before and after fitting a GPS-collar, to determine possible behavioural effects of wearing a collar. Although the study did not take place under ideal circumstances, indications of both behavioural, e.g. increased shaking behaviour, and physical, e.g. abrasions, effects of the collar were found. Even though our results were only based on two individuals, our findings stress the need for pilot studies in controlled environments before GPS collars to ensure safety of the study species and validity of the collected data.

Landscape variables affecting the Himalayan red panda Ailurus fulgens occupancy in wet season along the mountains in Nepal.

The Himalayan red panda is an endangered mammal endemic to Eastern Himalayan and South Western China. Data deficiency often hinders understanding of their spatial distribution and habitat use, which is critical for species conservation planning. We used sign surveys covering the entire potential red panda habitat over 22,453 km2 along the mid-hills and high mountains encompassing six conservation complexes in Nepal. To estimate red panda distribution using an occupancy framework, we walked 1,451 km along 446 sampled grid cells out of 4,631 grid cells in the wet season of 2016. We used single-species, single-season models to make inferences regarding covariates influencing detection and occupancy. We estimated the probability of detection and occupancy based on model-averaging techniques and drew predictive maps showing site-specific occupancy estimates. We observed red panda in 213 grid cells and found covariates such as elevation, distance to water sources, and bamboo cover influencing the occupancy. Red panda detection probability [Formula: see text] estimated at 0.70 (0.02). We estimated red panda site occupancy (sampled grid cells) and landscape occupancy (across the potential habitat) [Formula: see text] at 0.48 (0.01) and 0.40 (0.02) respectively. The predictive map shows a site-specific variation in the spatial distribution of this arboreal species along the priority red panda conservation complexes. Data on their spatial distribution may serve as a baseline for future studies and are expected to aid in species conservation planning in priority conservation complexes.

Fine-scale landscape genetics unveiling contemporary asymmetric movement of red panda (Ailurus fulgens) in Kangchenjunga landscape, India.

Wildlife management in rapid changing landscapes requires critical planning through cross cutting networks, and understanding of landscape features, often affected by the anthropogenic activities. The present study demonstrates fine-scale spatial patterns of genetic variation and contemporary gene flow of red panda (Ailurus fulgens) populations with respect to landscape connectivity in Kangchenjunga Landscape (KL), India. The study found about 1,309.54 km2 area suitable for red panda in KL-India, of which 62.21% area fell under the Protected Area network. We identified 24 unique individuals from 234 feces collected at nine microsatellite loci. The spatially explicit and non-explicit Bayesian clustering algorithms evident to exhibit population structuring and supported red panda populations to exist in meta-population frame work. In concurrence to the habitat suitability and landscape connectivity models, gene flow results supported a contemporary asymmetric movement of red panda by connecting KL-India in a crescent arc. We demonstrate the structural-operational connectivity of corridors in KL-India that facilitated red panda movement in the past. We also seek for cooperation in Nepal, Bhutan and China to aid in preparing for a comprehensive monitoring plan for the long-term conservation and management of red panda in trans-boundary landscapes.

Characteristics of Mesenchymal Stem Cells Isolated from the Bone Marrow of Red Pandas.

Mesenchymal stem cells (MSC) have strong therapeutic potential due to their capacity for self-renewal and multilineage differentiation. MSCs can also be useful in preserving the current genetic diversity of endangered wildlife. To date, MSCs from various species have been studied, but only a few species of endangered wild animals have been reported. Adult bone marrow (BM) is a rich source of mesenchymal stem cells. The aim of this study was to isolate and characterize MSCs derived from the BM of red pandas. Red panda BM-MSCs isolated from five individuals were fibroblast-like cells, similar to other species. Cultured BM-MSCs with normal karyotype were negative for the hematopoietic line marker CD34 and the endothelial cell marker CD31 but were positive for MSC markers, including CD44, CD105 and CD90. RT-PCR and western blot analysis showed self-renewal and pluripotency genes, including Oct4, Sox2 and Klf4, were also expressed in red panda BM-MSCs. Finally, red panda BM-MSCs had the potential for differentiation into osteogenic, adipogenic and neuron-like cells by using a combination of previously reported protocols for other species. We have therefore demonstrated that cells harvested from red panda bone marrow are capable of extensive in vitro multiplication and multilineage differentiation, which is an essential step toward their use in the preservation of red pandas biological diversity and future studies on MSC applications in endangered species.

Habitat overlap between Asiatic black bear Ursus thibetanus and red panda Ailurus fulgens in Himalaya.

Studying habitat overlap between sympatric species is one of the best ways to identify interspecies relationships and to direct conservation efforts so that multiple species can benefit. However, studies exploring interspecies relationships are very limited in Nepal, making it difficult for the government of Nepal and conservation partners to manage wildlife in their habitats, especially in Himalayan protected areas. In this study, we identified habitat overlap between Asiatic black bear (Ursus thibetanus) and red panda (Ailurus fulgens) as well as important habitat types for both species in the Makalu Barun National Park, Nepal using Maximum Entropy (MaxEnt) modeling. GPS points of species occurrence were collected from the field, and environmental variables were extracted from freely available sources. We found that the study area contained 647 km2 of Asiatic black bear habitat and 443 km2 of the red panda habitat. 368 km2 supported both species, which constituted 57% of the Asiatic black bear habitat and 83% of the red panda habitat. We found that conifer forest was the most important habitat type for both species. Because the largest portions of both species' habitat were located inside the buffer zone, a peripheral zone of national park, conservation efforts for these sympatric species should be focused inside the buffer zone to be most effective.

Potential Mechanism of Detoxification of Cyanide Compounds by Gut Microbiomes of Bamboo-Eating Pandas.

Gut microbes can enhance the ability of hosts to consume secondary plant compounds and, therefore, expand the dietary niche breadth of mammalian herbivores. The giant and red pandas are bamboo-eating specialists within the mammalian order Carnivora. Bamboo contains abundant plant secondary metabolites (e.g., cyanide-containing compounds). However, Carnivora species, including the giant panda, have deficient levels of rhodanese (one of the essential cyanide detoxification enzymes) in their tissues compared with the same tissues of herbivores. Here, we make a comparative analysis of 94 gut metagenomes, including 25 from bamboo-eating pandas (19 from giant pandas and 6 from red pandas), 30 from Père David's deer, and 39 from published data for other mammals. The bamboo-eating pandas' gut microbiomes had some common features, such as high proportions of Pseudomonas bacteria. The results revealed that bamboo-eating pandas' gut microbiomes were significantly enriched in putative genes coding for enzymes related to cyanide degradation (e.g., rhodanese) compared with the gut microbiomes of typical herbivorous mammals, which might have coevolved with their special bamboo diets. The enrichment of putative cyanide-digesting gut microbes, in combination with adaptations related to morphology (e.g., pseudothumbs) and genomic signatures, show that the giant panda and red panda have evolved some common traits to adapt to their bamboo diet.IMPORTANCE The giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens), two obligate bamboo feeders, have distinct phylogenetic positions in the order Carnivora. Bamboo is extraordinarily rich in plant secondary metabolites, such as allied phenolic and polyphenolic compounds and even toxic cyanide compounds. Here, the enrichment of putative cyanide-digesting gut microbes, in combination with adaptations related to morphology (e.g., pseudothumbs) and genomic signatures, show that the giant panda and red panda have evolved some common traits to adapt to their bamboo diet. Thus, here is another story of diet-driven gut microbiota in nature.

Current husbandry situation of red pandas in Japan.

The red panda (Ailurus fulgens) is an endangered semi-arboreal folivore with a specialized diet of bamboo leaves. Zoos worldwide maintain red pandas under a variety of housing environments and husbandry procedures that may affect the reproductive success and longevity of captive red pandas. The aims of the present study were to investigate how red pandas are kept in Japan and to obtain useful insights to increase their longevity by comparing the results from a questionnaire sent to Japanese zoos and those from a previous survey conducted in other countries. We received responses from 42 out of 52 zoos, holding a total of 219 individual red pandas, in Japan. The age-sex distribution indicated that red panda populations in Japan need close attention to be maintained sustainably. Of the husbandry aspects surveyed, the number of individuals in the same zoo, enclosure size, and the frequency of bamboo feeding factors indicated to affect reproductive success. Frequent monitoring of body weight, implementation of husbandry training, and bamboo feeding were also suggested as the reasons for the longevity of red pandas in Japan. Zoos were provided with sufficient physical features in on-exhibit enclosures, close monitoring for health status, and proper feeding, but other aspects of husbandry such as enclosure size and structures in off-exhibit spaces were insufficient and might compromise animal welfare.

Lineage-specific evolution of bitter taste receptor genes in the giant and red pandas implies dietary adaptation.

Taste 2 receptors (TAS2R) mediate bitterness perception in mammals, thus are called bitter taste receptors. It is believed that these genes evolved in response to species-specific diets. The giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens styani) in the order Carnivora are specialized herbivores with an almost exclusive bamboo diet (>90% bamboo). Because bamboo is full of bitter tasting compounds, we hypothesized that adaptive evolution has occurred at TAS2R genes in giant and red pandas throughout the course of their dietary shift. Here, we characterized 195 TAS2R genes in 9 Carnivora species and examined selective pressures on these genes. We found that both pandas harbor more putative functional TAS2R genes than other carnivores, and pseudogenized TAS2R genes in the giant panda are different from the red panda. The purifying selection on TAS2R1, TAS2R9 and TAS2R38 in the giant panda, and TAS2R62 in the red panda, has been strengthened throughout the course of adaptation to bamboo diet, while selective constraint on TAS2R4 and TAS2R38 in the red panda is relaxed. Remarkably, a few positively selected sites on TAS2R42 have been specifically detected in the giant panda. These results suggest an adaptive response in both pandas to a dietary shift from carnivory to herbivory, and TAS2R genes evolved independently in the 2 pandas. Our findings provide new insight into the molecular basis of mammalian sensory evolution and the process of adaptation to new ecological niches.

Large anthropogenic impacts on a charismatic small carnivore: Insights from distribution surveys of red panda Ailurus fulgens in Nepal.

Protected areas are key to preserving biodiversity and maintaining ecosystem services. However, their ability to ensure long-term survival of threatened andendangered species varies across countries, regions and landscapes. Distribution surveys can beparticularly important for assessing the value of protected areas, and gauging their efficacy incatering to species-specific requirements. We assessed the conservation value of one such reserve for a charismatic yet globally endangered species, the red panda Ailurus fulgens,in the light of on-going land-use transformation in Nepal. We conducted field surveys forindirect signs of red pandas along forest trails in 25-km2 sampling grid cells (n = 54) of Dhorpatan Hunting Reserve, and confronted a set of ecological hypotheses to the data using hierarchical occupancy models. We estimated overall occupancy at Ψ(SE) = 0.41 (0.007), with relatively high site-level detectability [p = 0.93 (SE = 0.001)]. Our results show that despitebeing a subsistence form of small-scale resource use, extraction of bamboo and livestock grazing negatively affected panda occurrence, albeit at different intensities. The amount of bamboo cover,rather than the overall proportion of forest cover, had greater influence on the panda occurrence. Despite availability of bamboo cover, areas with bamboo extraction and anthropogenic disturbances were less likely to be occupied by pandas. Together, these results suggest that long-term persistence of red pandas in this reserve and elsewhere across the species' range will require preventing commercial extractionof bamboo, coupled with case-specific regulation of anthropogenic exploitation of red panda habitats.

Integrating trans-abdominal ultrasonography with fecal steroid metabolite monitoring to accurately diagnose pregnancy and predict the timing of parturition in the red panda (Ailurus fulgens styani).

Red pandas (Ailurus fulgens styani) exhibit a variable gestation length and may experience a pseudopregnancy indistinguishable from true pregnancy; therefore, it is not possible to deduce an individual's true pregnancy status and parturition date based on breeding dates or fecal progesterone excretion patterns alone. The goal of this study was to evaluate the use of transabdominal ultrasonography for pregnancy diagnosis in red pandas. Two to three females were monitored over 4 consecutive years, generating a total of seven profiles (four pregnancies, two pseudopregnancies, and one lost pregnancy). Fecal samples were collected and assayed for progesterone (P4) and estrogen conjugate (EC) to characterize patterns associated with breeding activity and parturition events. Animals were trained for voluntary transabdominal ultrasound and examinations were performed weekly. Breeding behaviors and fecal EC data suggest that the estrus cycle of this species is 11-12 days in length. Fecal steroid metabolite analyses also revealed that neither P4 nor EC concentrations were suitable indicators of pregnancy in this species; however, a secondary increase in P4 occurred 69-71 days prior to parturition in all pregnant females, presumably coinciding with embryo implantation. Using ultrasonography, embryos were detected as early as 62 days post-breeding/50 days pre-partum and serial measurements of uterine lumen diameter were documented throughout four pregnancies. Advances in reproductive diagnostics, such as the implementation of ultrasonography, may facilitate improved husbandry of pregnant females and allow for the accurate prediction of parturition.

Mobility of the forearm in the raccoon (Procyon lotor), raccoon dog (Nyctereutes procyonoides) and red panda (Ailurus fulgens).

The ranges of pronation/supination of forearms in raccoons, raccoon dogs and red pandas were nondestructively examined. Three carcasses of each species were used for CT analysis, and the left forearms were scanned with a CT scanner in two positions: maximal supination and maximal pronation. Scanning data were reconstructed into three-dimensional images, cross-sectional images were extracted at the position that shows the largest area in the distal part of ulna, and then, the centroids of each cross section of the radius and ulna were detected. CT images of two positions were superimposed, by overlapping the outlines of each ulna, and then, the centroids were connected by lines to measure the angle of rotation, as an index of range of mobility. The measurements in each animal were analyzed, using the Tukey-Kramer method. The average angle of rotation was largest in raccoons and smallest in raccoon dogs, and the difference was significant. In the maximally pronated forearm of all species, the posture was almost equal to the usual grounding position with palms touching the ground. Therefore, the present results demonstrate that the forearms of raccoons can supinate to a greater degree from the grounding position with palms on the ground, as compared with those of raccoon dogs and red pandas.

Influence of climate on the survivorship of neonatal red pandas in captivity.

Red pandas, Ailurus fulgens, are popular exhibit animals in zoos. It is clear from data in the global studbook that there is considerable variation in their breeding success in different zoos. Population managers have long suspected that environmental temperature plays a key role in these differences. It is generally thought that this species, which is so well adapted to life in the cold damp climate of the mid-altitude forests of the Himalayas, has a problem coping with warmer climates. However, this hypothesis has not been tested until now. Using data extracted from the global studbook, we have demonstrated that climate at the location of birth has a clear impact on the survival of infant red pandas.

Diet and nutrient balance of red panda in Nepal.

We identified the winter plant species consumed by red panda in the Dhorpatan Hunting Reserve of eastern Nepal and compared this to the early-summer diet which was determined previously by Panthi et al. (2012). In addition, we estimated the proximate nutritional content of the leaves identified in red panda diet for both seasons, and we used nutritional geometry to explore macronutrient balance of leaves from the two different sampling periods. We identified six different plants in winter scats, which were the same as found in the previously determined early-summer diet. Arundinaria spp. bamboos were the main species found (82.1 % relative frequency), followed by Acer spp. (6.3 %), Betula utilis (4.6 %), Quercus semicarpifolia (3.7 %), Berberis spp. (1.3 %), and lichens (1.0 %), leaving 2.0 % unidentified. Geometric analysis suggested that the macronutrient balance of seasonal diets were similar in nutrient balance to the most frequently consumed Arundinaria spp. Differences in macronutrient balance may indicate seasonal nutrient preferences, such as increased carbohydrate intake in winter for thermogenesis, and increased protein and lipid intake in early summer to support reproduction and lactation; however, these differences may also indicate differences in resource availability. Habitat conserved for red panda in the region should include sufficient Arundinaria spp. as well as lesser consumed plants which may serve as complimentary foods.

Quantitative inferences on the locomotor behaviour of extinct species applied to Simocyon batalleri (Ailuridae, Late Miocene, Spain).

Inferences of function and ecology in extinct taxa have long been a subject of interest because it is fundamental to understand the evolutionary history of species. In this study, we use a quantitative approach to investigate the locomotor behaviour of Simocyon batalleri, a key taxon related to the ailurid family. To do so, we use 3D surface geometric morphometric approaches on the three long bones of the forelimb of an extant reference sample. Next, we test the locomotor strategy of S. batalleri using a leave-one-out cross-validated linear discriminant analysis. Our results show that S. batalleri is included in the morphospace of the living species of musteloids. However, each bone of the forelimb appears to show a different functional signal suggesting that inferring the lifestyle or locomotor behaviour of fossils can be difficult and dependent on the bone investigated. This highlights the importance of studying, where possible, a maximum of skeletal elements to be able to make robust inferences on the lifestyle of extinct species. Finally, our results suggest that S. batalleri may be more arboreal than previously suggested.

Three-dimensional computer simulations of feeding behaviour in red and giant pandas relate skull biomechanics with dietary niche partitioning.

The red (Ailurus fulgens) and giant (Ailuropoda melanoleuca) pandas are mammalian carnivores convergently adapted to a bamboo feeding diet. However, whereas Ailurus forages almost entirely on younger leaves, fruits and tender trunks, Ailuropoda relies more on trunks and stems. Such difference in foraging mode is considered a strategy for resource partitioning where they are sympatric. Here, we use finite-element analysis to test for mechanical differences and similarities in skull performance between Ailurus and Ailuropoda related to diet. Feeding simulations suggest that the two panda species have similar ranges of mechanical efficiency and strain energy profiles across the dentition, reflecting their durophagous diet. However, the stress distributions and peaks in the skulls of Ailurus and Ailuropoda are remarkably different for biting at all tooth locations. Although the skull of Ailuropoda is capable of resisting higher stresses than the skull of Ailurus, the latter is able to distribute stresses more evenly throughout the skull. These differences in skull biomechanics reflect their distinct bamboo feeding preferences. Ailurus uses repetitive chewing in an extended mastication to feed on soft leaves, and Ailuropoda exhibits shorter and more discrete periods of chomp-and-swallow feeding to break down hard bamboo trunks.

The use of the gonadotropin-releasing hormone analog deslorelin for short-term contraception in red pandas (Ailurus fulgens).

Red pandas (Ailurus fulgens) are threatened with extinction owing to habitat loss, exacerbated by their unique ecology and low fecundity. Regional breeding programs manage captive red panda populations. Recommendations not to breed may be made for various reasons, including genetic overrepresentation of certain individuals. No recommendations have been published on the use of contraception for red pandas. This article discusses the use of the GnRH analog deslorelin as a reversible method of contraception in both male and female pandas. The mean time from last contraception to conception was 3 years with a 4.6-mg deslorelin implant. The average dose of GnRH implant received was 1.09 mg/kg (range, 0.88-1.32). Males returned to breeding sooner than females. No reproductive side effects were noted with up to three consecutive annual GnRH implants.

Current husbandry of red pandas (Ailurus fulgens) in zoos.

The endangered red panda (Ailurus fulgens) is held in zoos worldwide. The aim of this study was to examine how red pandas are kept and managed in captivity and to compare it with the management guidelines. Sixty-nine zoos, mainly from Europe but also from North America and Australia/New Zealand, responded to our survey. The results revealed that in general zoos follow the management guidelines for most of the investigated issues. The average enclosure is almost four times larger than the minimum size recommended by the management guidelines, although seven zoos have smaller enclosures. About half the zoos do not follow the guidelines concerning visitor access and number of nest boxes. Other issues that may compromise animal welfare include proximity of neighboring carnivore species and placement of nest boxes.

Fibroblast cell line establishment, cryopreservation and interspecies embryos reconstruction in red panda ( Ailurus fulgens).

In evolution, the red panda (Ailurus fulgens) plays a pivotal role in the higher level phylogeny of arctoides carnivore mammals. The red panda inhabits certain Asian countries only and its numbers are decreasing. Therefore, the development of feasible ways to preserve this species is necessary. Genetic resource cryopreservation and somatic cell nuclear transfer (SCNT) have been used extensively to rescue this endangered species. The present study describes the establishment, for the first time, of a red panda ear fibroblast cell line, which was then cryopreserved, thawed and cultured. Through micromanipulation, interspecies embryos were reconstructed using the cryopreserved-thawed fibroblasts of the red panda as the donor and rabbit oocytes as recipients. A total of 194 enucleated rabbit oocytes were reconstructed with red panda ear fibroblasts; enucleated oocytes were activated without fusion as the control. The results show that the fibroblast cell line was established successfully by tissue culture and then cryopreserved in liquid nitrogen. Supplementation with 20% fetal bovine serum and 8% dimethyl sulphoxide in basic medium facilitated the cryopreservation. The interspecies embryos were successfully reconstructed. The cleavage, morulae and blastocyst rates after in vitro culture were 71, 47 and 23% (31/194), respectively. This study indicated that a somatic cell line could be established and cryopreserved from red panda and that rabbit cytoplast supports mitotic cleavage of the red panda karyoplasts and is capable of reprogramming the nucleus to achieve blastocysts.