Morphological examination of the visual system and orbital region in the red panda (Ailurus fulgens fulgens).

OBJECTIVES: The red panda is currently the only surviving member of the Ailuridae family in the Caniformia suborder. In this study, we provide data on anatomical, morphometric, histological and histochemical examination of the orbital region, eyelids, orbital gland, and eye tunics in two adult males Ailurus fulgens fulgens from the Wroclaw Zoological Garden, Poland. METHODS: The study involved morphometric analysis of the eyeball and selected accessory organs of the eye, along with analysis of the bony orbit, including its morphometry, macroscopic, and microscopic evaluation. Microscopic evaluation encompassed histological and histochemical staining, with the former involving hematoxylin & eosin (H&E), Movat pentachrome, picro-Mallory trichrome, Fontana-Masson, and the latter including PAS, AB pH 1.0, AB pH 2.5; AB pH 2.5/PAS, and HDI. RESULTS: The upper (UE) and lower (LE) eyelids presented well-developed tarsal glands, sebaceous glands, and a characteristic simple alveolar gland (producing a mucous secretion). The palpebral part of the lacrimal gland was present. A single lymphoid follicle was observed only in the upper eyelids. The superficial gland of the third eyelid (SGTE) was a multilobar acinar complex that produces mucous secretion and is contained within the interlobular ducts of numerous aggregates of lymph nodes. The third eyelid (TE) was T-shaped and composed of hyaline tissue, containing CALT. The lacrimal gland (LG) also revealed a multilobar acinar complex that produced mucous secretion, with a single lymphoid follicle. The cornea consisted of 4 layers, as Bowman's membrane was absent. The Vogt palisades composed of 7-10 layers of epithelial cells were demonstrated. The pupil was horizontally ovoid at rest (post-mortem). The sphincter pupil and the dilator pupil were well developed. Macroscopically, the tapetum lucidum appeared as a milky, non-opalescent crescent. Histologically, the choroidal tapetum lucidum cellulosum consisted of 5 to 9 layers of loosely packed oval cells. The retina showed a composition similar to that of terrestrial nocturnal carnivores. CONCLUSIONS: The results of our research indicate that the anatomical features of the eye and orbital region in the red panda share similarities with those described in the Musteloidea clade, as well as the Canidae and Ursidae families.

Red pandas with different diets and environments exhibit different gut microbial functional composition and capacity.

The red panda (Ailurus fulgens) is a distinctive mammal known for its reliance on a diet primarily consisting of bamboo. The gut microbiota and overall health of animals are strongly influenced by diets and environments. Therefore, conducting research to explore the taxonomical and functional variances within the gut microbiota of red pandas exposed to various dietary and environmental conditions could shed light on the dynamic complexities of their microbial communities. In this study, normal fecal samples were obtained from red pandas residing in captive and semi-free environments under different dietary regimes and used for metabolomic, 16S rRNA, and metagenomic sequencing analysis, with the pandas classified into four distinct cohorts according to diet and environment. In addition, metagenomic sequencing was conducted on mucus fecal samples to elucidate potential etiological agents of disease. Results revealed an increased risk of gastrointestinal diseases in red pandas consuming bamboo shoots due to the heightened presence of pathogenic bacteria, although an increased presence of microbiota-derived tryptophan metabolites appeared to facilitate intestinal balance. The red pandas fed bamboo leaves also exhibited a decrease in gut microbial diversity, which may be attributed to the antibacterial flavonoids and lower protein levels in leaves. Notably, red pandas residing in semi-free environments demonstrated an enriched gut microbial diversity. Moreover, the occurrence of mucus secretion may be due to an increased presence of species associated with diarrhea and a reduced level of microbiota-derived tryptophan metabolites. In summary, our findings substantiate the influential role of diet and environment in modulating the gut microbiota of red pandas, offering potential implications for improved captive breeding practices.

Amdoparvovirus-associated disease in red pandas (Ailurus fulgens).

The roster of amdoparvoviruses (APVs) in small carnivores is growing rapidly, but in most cases, the consequences of infection are poorly understood. Red panda amdoparvovirus (RPAV) is highly prevalent in zoo-housed red pandas and has been detected in both healthy and sick animals. Clarifying the clinical impact of RPAV in this endangered species is critical, and zoological collections offer a unique opportunity to examine viral disease association in carefully managed populations. We evaluated the potential impact of RPAV in captive red pandas with a combination of prospective and retrospective analyses. First, we collected feces from 2 healthy animals from one collection over a 6-year period and detected virus in 72/75 total samples, suggesting that RPAV can be a long-term subclinical infection. We next investigated the infections using a retrospective study of infection status and tissue distribution in a cohort of necropsied animals. We performed polymerase chain reaction and in situ hybridization on 43 necropsy cases from 4 zoo collections (3 from the United States, 1 from Europe, 1997-2022). RPAV was present in these populations for at least 2 decades before its discovery and is detectable in common and significant lesions of zoo-housed red pandas, including myocarditis (3/3 cases), nephritis (9/10), and interstitial pneumonia (2/4). RPAV is also detectable in sporadic lesions, including multisystemic pyogranulomatous inflammation, oral/pharyngeal mucosal inflammation, and dermatitis. The colocalization of virus with lesions supports a role in causation, suggesting that despite the apparently persistent and subclinical carriage of most infections, RPAV may have a significant impact in zoo collections.

Fatal cerebrovascular accident in a captive red panda (Ailurus fulgens fulgens) with concurrent amdoparvovirus infection.

We report the pathological and molecular findings in an adult male Himalayan red panda (Ailurus fulgens fulgens) whose death was attributed to parenchymal brain haemorrhage (PBH) of the thalamus. Post-mortem examination revealed severe, acute PBH and intraventricular haemorrhage with major involvement of the thalamus, as well as scattered chronic microinfarctions. Vascular disease in the brain and other organs was suggestive of systemic hypertension. Histological lesions included arteriolar hyalinosis and varying degrees of arteriosclerosis, arterial tunica media hypertrophy and hyperplasia and infiltration of arterial walls by lipid-laden macrophages. Other relevant findings included marked myocardial fibrosis, lymphoplasmacytic tubulointerstitial nephritis, lymphoplasmacytic meningoencephalitis and chronic mitral valve degeneration. The changes in the cerebral vasculature were consistent with hypertensive encephalopathy and a cerebrovascular accident, specifically PBH, which has not been previously reported in this species. Additionally, polymerase chain reaction analysis for red panda amdoparvovirus (RPAV) was positive in the brain and kidneys. Preceded by hypertensive vascular changes and brain microinfarctions, sudden death in this animal likely resulted from fatal PBH with intraventricular haemorrhage. The clinicopathological role of RPAV infection is unknown in this case, although its contribution to the chronic renal disease is considered possible in the context of our current understanding of RPAV-associated pathology.

Spatiotemporal walking gait kinematics of semi-arboreal red pandas (Ailurus fulgens).

Semi-arboreal mammals must routinely cope with the differing biomechanical challenges of terrestrial versus arboreal locomotion; however, it is not clear to what extent semi-arboreal mammals adjust footfall patterns when moving on different substrates. We opportunistically filmed quadrupedal locomotion (n = 132 walking strides) of semi-arboreal red pandas (Ailurus fulgens; n = 3) housed at Cleveland Metroparks Zoo and examined the effects of substrate type on spatiotemporal gait kinematic variables using linear mixed models. We further investigated the effects of substrate diameter and orientation on arboreal gait kinematics. Red pandas exclusively used lateral sequence (LS) gaits and most frequently utilized LS lateral couplet gaits across terrestrial and arboreal substrates. Red pandas moved significantly slower (p < 0.001), and controlling for speed, had significantly greater relative stride length (p < 0.001), mean stride duration (p = 0.002), mean duty factor (p < 0.001), and mean number of supporting limbs (p < 0.001) during arboreal locomotion. Arboreal strides on inclined substrates were characterized by significantly faster relative speeds and increased limb phase values compared with those horizontal and declined substrates. These kinematics adjustments help to reduce substrate oscillations thereby promoting stability on potentially precarious arboreal substrates. Red panda limb phase values are similar to those of (primarily terrestrial) Carnivora examined to date. Despite the similarity in footfall patterns during arboreal and terrestrial locomotion, flexibility in other kinematic variables is important for semi-arboreal red pandas that must navigate disparate biomechanical challenges inherent to arboreal versus terrestrial locomotion.

The complete mitochondrial genome of Ogmocotyle ailuri: gene content, composition and rearrangement and phylogenetic implications.

Trematodes of the genus Ogmocotyle are intestinal flukes that can infect a variety of definitive hosts, resulting in significant economic losses worldwide. However, there are few studies on molecular data of these trematodes. In this study, the mitochondrial (mt) genome of Ogmocotyle ailuri isolated from red panda (Ailurus fulgens) was determined and compared with those from Pronocephalata to investigate the mt genome content, genetic distance, gene rearrangements and phylogeny. The complete mt genome of O. ailuri is a typical closed circular molecule of 14 642 base pairs, comprising 12 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions. All genes are transcribed in the same direction. In addition, 23 intergenic spacers and 2 locations with gene overlaps were determined. Sequence identities and sliding window analysis indicated that cox1 is the most conserved gene among 12 PCGs in O. ailuri mt genome. The sequenced mt genomes of the 48 Plagiorchiida trematodes showed 5 types of gene arrangement based on all mt genome genes, with the gene arrangement of O. ailuri being type I. Phylogenetic analysis using concatenated amino acid sequences of 12 PCGs revealed that O. ailuri was closer to Ogmocotyle sikae than to Notocotylus intestinalis. These data enhance the Ogmocotyle mt genome database and provide molecular resources for further studies of Pronocephalata taxonomy, population genetics and systematics.

ECHOCARDIOGRAPHIC MEASUREMENTS IN ANESTHETIZED CAPTIVE RED PANDAS (AILURUS FULGENS).

A recent review of mortality in captive red pandas (Ailurus fulgens) listed cardiac pathology as the most common cause of death in the adult age group. The aim of this study was to describe the standard echocardiographic variables in 13 captive, healthy, adult red pandas that underwent an elective health examination. In addition, differences between the two subspecies of red panda, A. f. styani than in A. f. fulgens, and association of these echocardiographic variables with age, sex, and body condition score, were examined. Anesthesia was induced and maintained with isoflurane inhalant anesthetic. All animals underwent a full physical examination and comprehensive echocardiogram including 2D, M-mode, and Doppler modes of ultrasound examination. Mean and SD of the echocardiographic variables are reported. Systolic performance was considered subnormal due to the effect of the anesthetic agent. The echocardiographic variables were similar for the two subspecies and sexes, with the exception of the left atrial dimension (2D), which measured larger (P= 0.03) in A. f. styani than in A. f. fulgens, and left ventricular internal diameter in diastole, which measured larger (P = 0.04) in males than females. Several echocardiographic measurements were correlated with age (P < 0.05), whereas the only measurement that significantly correlated with body condition score was end diastolic volume (P = 0.01). These results provide ranges that can offer guidance for predicting cardiac disease in red panda.

Gene expressions between obligate bamboo-eating pandas and non-herbivorous mammals reveal converged specialized bamboo diet adaptation.

BACKGROUND: It is inevitable to change the function or expression of genes during the environmental adaption of species. Both the giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens) belong to Carnivora and have developed similar adaptations to the same dietary switch to bamboos at the morphological and genomic levels. However, the genetic adaptation at the gene expression level is unclear. Therefore, we aimed to examine the gene expression patterns of giant and red panda convergent specialized bamboo-diets. We examined differences in liver and pancreas transcriptomes between the two panda species and other non-herbivorous species. RESULTS: The clustering and PCA plots suggested that the specialized bamboo diet may drive similar expression shifts in these two species of pandas. Therefore, we focused on shared liver and pancreas DEGs (differentially expressed genes) in the giant and red panda relative to other non-herbivorous species. Genetic convergence occurred at multiple levels spanning carbohydrate metabolism, lipid metabolism, and lysine degradation. The shared adaptive convergence DEGs in both organs probably be an evolutionary response to the high carbohydrate, low lipid and lysine bamboo diet. Convergent expression of those nutrient metabolism-related genes in both pandas was an intricate process and subjected to multi-level regulation, including DNA methylation and transcription factor. A large number of lysine degradation and lipid metabolism related genes were hypermethylated in promoter regions in the red panda. Most genes related to carbohydrate metabolism had reduced DNA methylation with increased mRNA expression in giant pandas. Unlike the red panda, the core gene of the lysine degradation pathway (AASS) doesn't exhibit hypermethylation modification in the giant panda, and dual-luciferase reporter assay showed that transcription factor, NR3C1, functions as a transcriptional activator in AASS transcription through the binding to AASS promoter region. CONCLUSIONS: Our results revealed the adaptive expressions and regulations of the metabolism-related genes responding to the unique nutrients in bamboo food and provided data accumulation and research hints for the future revelation of complex mechanism of two pandas underlying convergent adaptation to a specialized bamboo diet.

Comparative Transcriptomics and Methylomics Reveal Adaptive Responses of Digestive and Metabolic Genes to Dietary Shift in Giant and Red Pandas.

Both the giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens) belong to the order Carnivora, but have changed their dietary habits to eating bamboo exclusively. The convergent evolution characteristics of their morphology, genome and gut flora have been found in the two pandas. However, the research on the convergent adaptation of their digestion and metabolism to the bamboo diet, mediated by the dietary shift of the two pandas at the gene-expression and epigenetic regulation levels, is still lacking. We therefore used RNA sequencing among five species (two pandas and three non-herbivore mammals) and bisulfite sequencing among three species (two pandas and a carnivore ferret) to sequence key digestion and metabolism tissues (stomach and small intestine). Our results provide evidence that the convergent differentially expressed genes (related to carbohydrate utilization, bile secretion, Lys and Arg metabolism, vitamin B12 utilization and cyanide detoxification) of the two pandas are adaptive responses to the bamboo diet containing low lipids, low Lys and Arg, low vitamin B12 and high cyanide. We also profiled the genome-wide methylome maps of giant panda, red panda and ferret, and the results indicated that the promoter methylation of the two pandas may regulate digestive and metabolic genes to adapt to sudden environmental changes, and then, transmit genetic information to future generations to evolve into bamboo eaters. Taken together, our study provides new insights into the molecular mechanisms of the dietary shift and the adaptation to a strict bamboo diet in both pandas using comparative transcriptomics and methylomics.

Predictors of psychological stress and behavioural diversity among captive red panda in Indian zoos and their implications for global captive management.

At high elevations, biodiversity is at elevated risk from extinctions due to rapid environmental changes. In the most of its range in Himalayas, the red panda, an endangered species, is struggling to survive in the wild, and a global captive breeding programme has been launched to conserve the species. Because captivity can have negative impacts on animals, reducing the chance of successful reintroduction, we investigated the predictors of stereotyped behaviour and behavioural diversity of red panda (n = 26), and the effect of stereotypy on their behavioural diversity in three Indian zoos. Multivariate analysis showed that stereotypy increased with density of logs on the ground, age and higher among pandas in zoo 3 compared to zoo 2, but decreased with number of nests, sociality, tree density and tree height used. Similarly, behavioural diversity increased with log density, but decreased among pandas in zoo 2 compared to zoo 1, during summer compared to winter, and also with ambient temperature, stereotypy, tree density, and tree height used. The relationship between stereotypy and behavioural diversity was negative, but not significant. Provision of a greater density of trees was associated with higher levels of daytime inactivity. Findings from this study have global relevance, as the captive red panda experiences similar welfare issues around the world, and our data provide empirical support for some existing guidelines for red panda husbandry.

Food handling shapes the laterality of paw use in the Chinese red panda (Ailurus styani).

Limb preference, associated with behavioural laterality and hemispheric lateralization, has long received high attention from both ethologists and psychologists. Chinese red pandas in Shanghai Zoo perform natural unimanual actions in two different ways during food handling, making it an appropriate subject to investigate the effect of food handling on pawedness. In this study, we estimated the direction and the strength of pawedness in spontaneous feeding behaviours in zoo-housed Chinese red pandas. All the subjects exclusively used the preferred paw when handling fruit pieces throughout the observational period, with three of them preferred the left paw while the other two the right paw. By contrast, both paws were found to be used alternately in consuming bamboo. Our results indicate that different food manipulations have effects on pawedness in the Chinese red panda where more complex handling could probably enhance its pawedness.

AMDOPARVOVIRUS INFECTIONS ARE PREVALENT, PERSISTENT, AND GENETICALLY DIVERSE IN ZOO-HOUSED RED PANDAS (AILURUS FULGENS).

Red pandas (Ailurus fulgens) are a globally endangered small carnivoran species and subjects of a robust ex situ conservation effort that includes animals housed in zoos. In 2018, red panda amdoparvovirus (RPAV) was discovered by metagenomics analyses of tissues from two geriatric red pandas, and in one case it was associated with significant lesions. Because RPAV was discovered in a single zoo cohort, it was unclear whether these infections represented a widely distributed, enzootic virus of red pandas or a localized 'spillover' from a different host species into this collection. The first goal of this study was to estimate the prevalence of RPAV in US zoos. The authors amplified RPAV from feces of 104 individual red pandas from 37 US zoos, and the virus was detected in 52/104 samples (50.0%). Next, to establish persistence of infection in individual animals, the authors tested serial samples in a single cohort over a 4.5-yr period, and virus was consistently shed by infected animals throughout the sampling period. Finally, full viral coding sequences were amplified and sequenced from three cases, and partial sequences of both the nonstructural and capsid genes were obtained for an additional 19 cases. RPAV is a genetically diverse but monophyletic viral species, and multiple viral lineages are present in US zoo-housed red pandas. The authors do not know how red pandas were originally infected, but RPAV is very common in red pandas in the United States, and infections are persistent-presumably for the lifetime of the animal.

Mortality analysis of captive red panda cubs within Chengdu, China.

BACKGROUND: The red panda has been classified as an endangered species due to the decreased number in the world and disease is considered as a great threat to the health and survival of the cubs in captivity. RESULTS: This study analyzed 32 red panda cub mortalities (15 females and 17 males, age less than two months) through gross necropsy, microbiological examination, and histopathological observation at the Chengdu Research Base of Giant Panda Breeding, China, during 2014-2020. The results showed that screenings for canine distemper virus, canine parvovirus, rotavirus and parasite infection were all negative, however bacteria such as Klebsiella pneumoniae, Proteus mirabilis, Escherichia coli, Enterococcus faecalis, Pseudomonas were isolated from the tissue samples of some cubs. The major causes of death were respiratory (43.75%) and digestive system disease (28.13%), followed by cardiovascular disease (12.5%) and neonatal stillbirths (9.38%). Renal system diseases and trauma were also detected, at lower incidence (one case for each). The mortality rate within 15 days of birth was 68.75% and gradually decreased with age, there was no significant difference in gender. CONCLUSION: This study can provide a scientific basis for the analysis of the cause of death among red panda cubs in captivity, so as to improve the survival rate, help build the captive population and further the ex-situ conservation management of this endangered species. Additionally, our research may also provide insights into the in-situ conservation of wild red pandas by identifying emerging disease threats within the wild population and potential treatment for rescued individuals.

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.

Red Panda feces from Eastern Himalaya as a modern analogue for palaeodietary and palaeoecological analyses.

Modern feces samples of the endangered red panda (Ailurus fulgens) were examined using multiproxy analysis to characterize the dietary patterns in their natural habitat in India. An abundance of Bambusoideae phytoliths and leaves (macrobotanical remains) provide direct evidence of their primary dietary plants. In contrast, Bambusoideae pollen is sporadic or absent in the pollen assemblages. An abundance of Lepisorus spores and its leaves along with broadleaved taxa, Betula, Engelhardtia, and Quercus are indicative of other important food sources. Average δ13C values (- 29.6‰) of the red panda feces indicate typical C3 type of plants as the primary food source, while the, δ15N values vary in narrow range (3.3-5.1‰) but conspicuously reveal a seasonal difference in values most likely due to differing metabolic activities in summer and winter. The multiproxy data can provide a baseline for the reconstruction of the palaeodietary and palaeoecology of extinct herbivores at both regional and global scales.

PRESUMPTIVE CONGENITAL HYPOTHYROIDISM IN RED PANDAS (AILURUS FULGENS FULGENS) FROM FOUR SUCCESSIVE LITTERS.

High neonatal mortality among red pandas (Ailurus fulgens) challenges the long-term sustainability of the Species Survival Plan population. Congenital hypothyroidism (CH) is a rare condition in domestic animals, typically due to an inherited genetic defect. Nongoitrous CH was presumptively diagnosed in 75% (n = 6/8) of red panda neonates from four successive litters, with a common sire and two closely related dams. Antemortem diagnosis of CH was made in three cubs (n = 3/6) based on elevated thyroid stimulating hormone and decreased free thyroxine and total thyroxine levels. Affected cubs also had suggestive clinical signs, which included delayed growth with cretinous dwarf appearance, atonic bladder, delayed gastrointestinal motility, hypercholesterolemia, and hypocalcemia. With sodium levothyroxine therapy, two of the three cubs developed into normal adult red pandas in terms of body size, appearance, and behavior. On necropsy cubs (n = 4) were small with varying degrees of cretin dwarf appearance and hypoplastic thyroids with reduced to no colloid in follicles. These cases demonstrate the importance of collecting thyroid tissue, (or proximal trachea/larynx if gross visualization not possible), in neonates for histopathology. Further investigation into the role of thyroid disease in neonatal red panda mortality is warranted.

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.

A tail of two pandas- whole genome k-mer signature analysis of the red panda (Ailurus fulgens) and the Giant panda (Ailuropoda melanoleuca).

BACKGROUND: The red panda (Ailurus fulgens) is a riddle of morphology, making it hard to tell whether it is an ursid, a procyonid, a mustelid, or a member of its own family. Previous genetic studies have given quite contradictory results as to its phylogenetic placement. RESULTS: A recently developed whole genome-based algorithm, the Whole Genome K-mer Signature algorithm was used to analyze the genomes of 28 species of Carnivora, including A. fulgens and several felid, ursid, mustelid, one mephitid species. This algorithm has the advantage of holistically using all the information in the genomes of these species. Being a genomics-based algorithm, it also reduces stochastic error to a minimum. Besides the whole genome, the mitochondrial DNA from 52 mustelids, mephitids, ursids, procyonids and A. fulgens were aligned to draw further phylogenetic inferences. The results from the whole genome study suggested that A. fulgens is a member of the mustelid clade (p = 9·10- 97). A. fulgens also separates from the mephitid Spilogala gracilis. The giant panda, Ailuropoda melanoleuca also clusters away from A. fulgens, together with other ursids (p = 1.2·10- 62). This could be due to the geographic isolation of A. fulgens from other mustelid species. However, results from the mitochondrial study as well as neighbor-joining methods based on the sequence identity matrix suggests that A. fulgens forms a monophyletic group. A Maximum Likelihood tree suggests that A. fulgens and Ursidae form a monophyletic group, although the bootstrap value is weak. CONCLUSIONS: The main conclusion that we can draw from this study is that on a whole genome level A. fulgens possibly belongs to the mustelid clade, and not an ursid or a mephitid. This despite the fact that previously some researchers classified A. fulgens and A. melanoleuca as relatives. Since the genotype determines the phenotype, molecular-based classification takes precedence over morphological classifications. This affirms the results of some previous studies, which studied smaller portions of the genome. However, mitochondrial analyses based on neighbor-joining and maximum likelihood methods suggest otherwise.

Reaching over the gap: A review of trends in and status of red panda research over 193 years (1827-2020).

The red panda is a unique species taxonomically known for its peculiar biological and ecological characteristics, and extreme attractiveness. Despite being highly significant from conservation, scientific and economic perspectives, this species has experienced a declining population in the wild. Thus, to direct further research priorities and conservation actions and assess gaps in the current research trend of this species, a systematic literature review was conducted covering 175 journal articles published in English over 193 years (1827-2020). This review revealed that (1) the biological aspect was highly studied compared to other thematic areas of red panda (2) captive-based studies are relatively higher than the studies based in wild populations (3) China is leading the red panda studies amongst all red panda range (4) The universities were found contributing more to red panda studies than other institutions. Surprisingly, we found that the researchers from the non-range country were leading red panda study than those from range countries. Our review highlighted the need of prioritising studies in underrepresented locations and understudied thematic areas focusing on the assessment of climate change impact, bamboo distribution status, ecosystem services of red panda habitat, behavior and movement ecology, population estimation, and metapopulation dynamics. We urge landscape-level studies and long-term population monitoring. Besides, we also suggest the documentation and evaluation of the effectiveness of ongoing red panda-focused conservation programs. We also stress the need for strengthening the capacity of institutions and people from range countries.

Genetic assessment of captive red pandas (Ailurus fulgens) in American zoos to address management separation by putative subspecies.

Red pandas (Ailurus fulgens) are small charismatic mammals native across montane southern Asia, now endangered by human impacts. They are considered "living fossils" as the sole member of a distinct family, warranting higher conservation priority. Therefore, ex situ breeding programs were initiated to prevent extinction and act as genetic reservoirs for reintroduction, although complicated by apparent taxonomic subdivision. This study investigated whether the separation of captive red pandas in the North American Species Survival Plan® by putative subspecies was justified. A 383-bp segment of mitochondrial DNA control region was therefore sequenced from 67 members representing matriarchal lineages of both groups. A network analysis placed the 11 haplotypes found into separate but closely connected clusters, with one group more strongly related than the other. Statistical analyses and diversity indices corroborated differentiation between the two management units. Phylogenetic analyses employing multiple outgroups confirmed, although not robustly, reciprocal monophyly of the four- and seven-haplotype clades representing putative subspecies Ailurus fulgens fulgens and Ailurus fulgens styani, respectively. These empirical results are adequate to justify continued independent management of these zoo subpopulations, but cannot be definitive for taxonomic classification due to limited sampling from their native range. They will, however, be useful in evaluating long-term genetic diversity changes, focusing management efforts on newly revealed evolutionary limitations, and comparing with an assessment of wild red pandas to determine how representative zoo populations are for reintroduction purposes. Maintaining genetic diversity and population structure of endangered species is essential to protect evolutionary potential and adaptations for long-term sustainability.