Panda Research Articles

Feline Panleukopenia Virus With G299E Substitution in the VP2 Protein First Identified From a Captive Giant Panda in China.

A feline panleukopenia virus (FPV), Giant panda/CD/2018, was isolated from a captive giant panda with mild diarrhea in 2018 in Chengdu, China, and further identified via indirect immunofluorescence assay (IFA), transmission electron microscopy (TEM) observation, and genetic analysis. Phylogenetic analysis based on the complete VP2 nucleotide sequences showed that it shared high homology with Chinese FPV isolates and grouped within FPV cluster 1. One unique substitution Gly(G)299Glu(E) in the capsid protein VP2 was first identified with Giant panda/CD/2018. The presence of the G299E substitution is notable as it is located on the top region of the interconnecting surface loop 3, which may be involved in controlling the host range and antigenicity of FPV. These findings first demonstrate that FPV with natural point mutation G299E in the VP2 gene is prevalent in giant panda and suggest that etiological surveillance and vaccination among all giant pandas are urgently needed to protect this endangered species against FPV infection.

Medical management of open pyometra in a giant panda (Ailuropoda melanoleuca)

AbstractThe female giant panda (Ailuropoda melanoleuca) has a characteristic reproductive physiology that involves a defined breeding season, embryonic diapause and potential for pseudopregnancy. The hormonal variation related to these reproductive characteristics may be a factor contributing to reproductive pathologies. In this case report, open pyometra was diagnosed in a giant panda. Clinical signs included lethargy, anorexia and mucopurulent vulvar discharge. Diagnostic tests performed included complete blood cell count, plasma biochemistry, protein electrophoresis, cytology, culture and sensitivity and abdominal ultrasonography. Treatment consisted of systemic antibiotics based on culture and sensitivity. Response to treatment was monitored via ultrasonography and visual examination of the vulvar region. Two weeks after treatment, the pyometra resolved and no subsequent complications were seen. This is the first case of pyometra reported in a giant panda. Pyometra should be considered as a differential diagnosis in entire female giant pandas presenting with abnormal clinical signs following breeding season.

High Prevalence of Antimicrobial Resistance and Integron Gene Cassettes in Multi-Drug-Resistant Klebsiella pneumoniae Isolates From Captive Giant Pandas (Ailuropoda melanoleuca).

Multi-drug-resistant Klebsiella pneumoniae (MDR K. pneumonia) is increasingly being reported with corresponding increase in morbidity and mortality all over the world. However, limited information is available concerning MDR K. pneumonia in giant pandas. The objective of this study was to grasp the drug resistance profile of MDR K. pneumonia isolated from giant pandas. A total of 182 K. pneumoniae isolates were collected from fresh feces of 94 captive giant pandas of different ages and sex and separated by season. We performed a standard disk diffusion antimicrobial susceptibility test with the isolates and further evaluated the antibiotic resistance genes (ARGs) of multi-drug-resistant strains by high-throughput quantitative PCR. In addition, we then analyzed mobile genetic elements (MGEs), integron gene cassettes, and the multi-locus sequence typing of multi-drug-resistant strains by PCR. Antimicrobial susceptibility testing results demonstrated that a total of 30 (16.5%) K. pneumoniae isolates showed multiple drug resistances. The thirty MDR K. pneumonia isolates were mainly resistant to amoxicillin (100.0%), doxycycline (86.7%), chloramphenicol (60.0%), compound trimethoprim (60.0%) and trimethoprim (56.7%). Fifty different types of antibiotic resistance genes were found, which included a total of 671 antibiotic resistance genes, in the 30 multi-drug-resistant isolates. The top ten resistance genes were: vanTC-02, aacC, blaCTX-M-04, blaSHV-01, blaSHV-02, ampC-04, blaOXY, tetD, blaTEM and tetA-02. Thirteen mobile genetic elements were detected, of which IS26 (96.67%) and intI1 (96.67%) had the highest frequency. The thirty MDR K. pneumonia isolates were negative for the traA, traF, tnsA, IS1133, ISpa7, ISkpn6, intI2 and intI3 genes. Moreover, a further investigation of integrons revealed that two types of specific gene cassettes (dfrA12 + orfF + aadA2 and dfrA12 + orfF) were identified in class 1 integrons. Multi-locus sequence typing results showed that 22 STs in the thirty MDR K. pneumonia isolates were identified, the main type was ST37 (5/30). Our results illustrate that effective surveillance and strict biosecurity strategies should be taken to prevent the spread of multi-drug-resistant bacteria, and monitor the emergence of mobile genetic elements and integrons.

Identifying core habitats and corridors for giant pandas by combining multiscale random forest and connectivity analysis.

Habitat loss and fragmentation are widely acknowledged as the main driver of the decline of giant panda populations. The Chinese government has made great efforts to protect this charming species and has made remarkable achievements, such as population growth and habitat expansion. However, habitat fragmentation has not been reversed. Protecting giant pandas in a large spatial extent needs to identify core habitat patches and corridors connecting them. This study used an equal‐sampling multiscale random forest habitat model to predict a habitat suitability map for the giant panda. Then, we applied the resistant kernel method and factorial least‐cost path analysis to identify core habitats connected by panda dispersal and corridors among panda occurrences, respectively. Finally, we evaluated the effectiveness of current protected areas in representing core habitats and corridors. Our results showed high scale dependence of giant panda habitat selection. Giant pandas strongly respond to bamboo percentage and elevation at a relatively fine scale (1 km), whereas they respond to anthropogenic factors at a coarse scale (≥2 km). Dispersal ability has significant effects on core habitats extent and population fragmentation evaluation. Under medium and high dispersal ability scenarios (12,000 and 20,000 cost units), most giant panda habitats in the Qionglai mountain are predicted to be well connected by dispersal. The proportion of core habitats covered by protected areas varied between 38% and 43% under different dispersal ability scenarios, highlighting significant gaps in the protected area network. Similarly, only 43% of corridors that connect giant panda occurrences were protected. Our results can provide crucial information for conservation managers to develop wise strategies to safeguard the long‐term viability of the giant panda population.

Bear diets and human–bear conflicts: insights from isotopic ecology

AbstractBears, Ursidae, are considered omnivores, except for giant pandas Ailuropoda melanoleuca and polar bears Ursus maritimus. However, omnivory includes a wide range of dietary variation and trophic positions, making bear dietary ecology unclear. We inferred bear trophic positions from δ15N (‰) values and examined their correlation with diets reported in the literature, including frequency of human–bear conflicts (livestock predation and crop damage incidents). Overall, 15N signatures were consistent with diet estimates. Bear species with higher 15N signatures differed more from each other, including cases of large regional intraspecific variance, than bear species with lower 15N signatures. Bear trophic position and frequency of reports of human–bear conflicts were uncorrelated, suggesting that livestock predation by bears is an opportunistic behaviour rather than a response to food availability dynamics.

Sesquiterpenoids isolated from feces-residing Streptomyces sp. inhibit the cellular entry of influenza a viruses

ABSTRACT: Five metabolites (1–5), including two new sesquiterpenoids, designated ganodermanol L (1) and 4α,15-epoxyeudesmane-1β,6α,11-triol (2), together with three known structurally related compounds (3-5), have been isolated from the cultures of Streptomyces sp. XM17, a bacteria residing in the fresh feces of the giant panda Ailuropoda melanoleuca. The structures of 1–2 were established on the basis of extensive spectroscopic analyses, including 1D- and 2D-NMR (1H-1H COSY, HMQC, HMBC and NOESY) experiments. Furthermore, the absolute configuration of 1 was established by single-crystal X-ray crystallographic analyses. Of noted, these compounds were found to possessed antiviral activities using the ‘pretreatment of virus’ approach with IC50 values ranging from 4 to 30 nM, indicating that these sesquiterpenoids were potent in inhibiting the entry of influenza A virus.

Consequences of nescient mating: Artificial insemination increases cub rejection in the giant panda (Ailuropoda melanoleuca)

Artificial insemination (AI) is a valuable tool for conservation breeding programs as it can promote reproductive success and genetic diversity in species with decreasing populations. However, females that undergo AI do not experience natural mating events, which we term, “nescient mating.” Thus, these females would not be able to assess the quality of their mate through physical, olfactory, or visual cues. Previous research has found that AI can lead to diminished breeding outcomes when compared to natural mating. In this study, we investigated whether fertilization method (natural mating versus AI) affects postpartum behaviors, such as cub rejection and maternal care in female giant pandas. We analyzed data from 202 cubs and 43 mother-cub dyads between 1996 and 2018. We did not find a difference in gestation length, cub weight, or cub survivorship between naturally mated and artificially inseminated females. Compared to naturally inseminated females, artificially mated females were significantly more likely to reject cubs (p = 0.039). Contrary to what we had predicted, maternal care and infant behaviors during the first month did not differ between natural mating and AI groups. Taken together, our results indicate that female giant pandas who experienced nescient mating may have reduced offspring investment, manifest in increased cub rejection. However, the lack of behavioral differences between AI and natural mating groups was encouraging, indicating that once a female accepted a cub to rear, maternal care was adequate to ensure cub survival. Nonetheless, we recommend that conservation breeding programs prioritize natural mating as higher rates of maternal rejection increase the facility and human time cost of cub rearing and also the chances of detrimental downstream effects on offspring adult behaviors in the giant panda.

Organic Fertilizers Shape Soil Microbial Communities and Increase Soil Amino Acid Metabolites Content in a Blueberry Orchard.

The decline in soil nutrients is becoming a major concern of soil degradation. The possibility of using organic waste as a soil additive to increase nutrients and essential components is significant in soil quality protection and waste management. The aim of this study was to investigate the effects of composted spent mushroom substrate (MS), giant panda feces (PF), and cattle manure (CM) as organic fertilizers in soil microbial communities and metabolites in blueberry orchard in China, which were measured by using high-throughput sequencing and gas chromatography-mass spectrometry (GC-MS)-based metabolomics. Altogether, 45.66% of the bacterial operational taxonomic units (OTUs) and 9.08% of the fungal OTUs were detected in all treatments. Principal coordinates analysis demonstrated that the bacterial and fungal communities in MS and PF treatments were similar, whereas the communities in the not-organic fertilized control (CK) were significantly different from those in the organic fertilizer treatments. Proteobacteria, Acidobacteria, and Bacteroidetes were the dominant bacterial phyla, and Basidiomycota, Ascomycota, and Mortierellomycota the dominant fungal phyla. Redundancy analysis indicated that pH and available potassium were the main factors determining the composition of microbial communities. The fungal genera Postia, Cephalotrichum, and Thermomyces increased in organic fertilizer treatments, and likely promoted the degradation of organic fertilizers into low molecular-weight metabolites (e.g., amino acids). PCA and PLS-DA models showed that the metabolites in CK were different from those in the other three treatments, and those in CM were clearly different from those in MS and PF. Co-occurrence network analysis showed that several taxa correlated positively with amino acid contents. The results of this study provide new insights into organic waste reutilization and new directions for further studies.

Viral metagenomics unveiled extensive communications of viruses within giant pandas and their associated organisms in the same ecosystem

Cross-species transmission events were commonplace, with numerous cases of host-switching during the viral evolutionary history, but relatively little evidence for onward transmission in different species living in the same ecosystem. For understanding the communications of viruses in giant pandas (Ailuropoda melanoleuca) and their associated organisms, based on a large size of samples (N = 2305) collected between 2015 and 2020 from giant panda (N = 776) and other four giant panda-associated organisms in the same ecosystem, red pandas (N = 700), stray cats (N = 32), wild rats (N = 42), and mosquitoes (N = 755), viromics was used for the virus identification and subsequent virus traceability. The results showed that a feline panleukopenia virus (FPV) was found in giant pandas with clinical signs of vomiting and mild diarrhea. Meanwhile, the same FPV strain was also prevalent in the healthy red panda (Ailurus fulgens) population. From the viromes of the five different organisms, 250 virus genomes were determined. Our data revealed that besides FPV, other putative pathogenic viruses, such as red panda amdoparvoviruses (RPAVs) and Getah viruses (GETVs) were responsible for previous disease or death of some red pandas. We also demonstrated that a number of viruses were involved in potential interspecies jumping events between giant pandas and their associated species. Collectively, our results shed light on the genetic diversity and relationship of diverse viral pathogens in ‘Giant pandas-Associated animals-Arthropods’ and report some cases of possible viral host-switching among these host species living in the same ecosystem.

Back-and-forth shifts in habitat selection by giant pandas over the past two decades in the Daxiangling Mountains, southwestern China

Habitat selection is a complex process, and animals may change their habitat selection over time. However, we have a poor understanding of temporal variation in habitat selection of endangered giant pandas (Ailuropoda melanoleuca), which has limited the development of conservation strategies for them. This study examined giant panda habitat selection in three stages of the past two decades (2001–2002, 2011–2012, and 2019–2020) in the Daxiangling Mountains, southwestern China. We applied the resource selection function to estimate the probability of species occurrence with five limiting habitat factors, i.e., elevation, slope, aspect, tree diameter at breast height, and bamboo cover. We found that giant pandas consistently selected for south- and west-facing slopes with mid-elevations and large trees in the three stages. However, there were shifts back-and-forth in the choice between higher and lower elevations and between higher and lower bamboo cover across the three stages. We suggest that the giant panda habitat selection had important consequences for gaining the high availability of palatable bamboo and good shelters. The back-and-forth shifts might be the result of changes in the type, spatial distribution, and intensity of human disturbance. Our study provides insights for improving our understanding and approaches for quantifying temporal variation in habitat selection in giant pandas and other species, suggesting that adaptive conservation and management in response to changing environments is needed.

Multi-functional Potential of Five Lactic Acid Bacteria Strains Derived from Giant Panda (Ailuropoda melanoleuca).

The multi-functional properties of lactic acid bacteria (LAB) on host health have been a popular research topic. The aim of present study was to assess the multi-functional potential of five LAB strains isolated from giant panda. In this study, we analyzed five giant panda LAB strains (Weissella confuse WJ202003 (W3), WJ202009 (W9), WJ202021 (W21), BSP201703 (X3); Lactiplantibacillus plantarum BSGP201683 (G83)) and found that they exhibited rapid growth as well as strong acid production capacity. The five LAB strains possessed high cell surface hydrophobicity to the four tested solvents (xylene, hexadecane, chloroform, ethyl acetate; except strain W9), auto-aggregation ability, co-aggregation ability with three pathogens (Escherichia coli, Enterotoxigenic Escherichia coli, Salmonella), adhesion ability to Caco-2 cell line, and strongly biofilm formation ability, suggesting an adhesion property. As investigated for their antioxidative potential, all the strains showed good tolerance to H2O2, high scavenging ability against 1, 1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl (OH-), and reduction ability. Furthermore, the five LAB strains could produce multiple probiotic substances, including exopolysaccharide (EPS), gamma-aminobutyric acid (GABA), bile salt hydrolase (BSH), cellulase (only strain G83), and protease (except strain X3), which was the first to report the production of EPS, GABA, BSH, cellulase, and protease in giant panda-derived LAB strain. These results demonstrated that strains W3, W9, W21, X3, and G83 had multi-functional potential and could be utilized as potential probiotics for giant panda.

Resveratrol and lycium barbarum polysaccharide improve Qinling giant panda (Ailuropoda melanoleuca Qinlingensis) sperm quality during cryopreservation

BackgroundSemen cryopreservation has become an essential tool for conservation efforts of the giant panda (Ailuropoda melanoleuca); however, it is severely detrimental to sperm quality. Evidence has shown that antioxidants have the potential to reverse cryopreservation-induced damage in sperm. The purpose of this study was to screen effective antioxidants that could retain sperm quality during cryopreservation and to determine the optimal dose. Seven antioxidant groups, including resveratrol (RSV = 50 μM, RSV = 100 μM, RSV = 150 μM), lycium barbarum polysaccharide (LBP = 2 mg/mL, LBP = 4 mg/mL), laminaria japonica polysaccharides (LJP = 1 mg/mL) or combination (LBP = 2 mg/mL, LJP = 1 mg/mL and RSV = 100 μM) were assessed.ResultsRSV, LBP, LJP, or a combination of RSV, LBP, and LJP added to the freezing medium significantly improved sperm progressive motility, plasma membrane integrity, acrosome integrity, and mitochondrial activity during the cryopreservation process. Furthermore, the activities of glutathione peroxidase and superoxide dismutase were also improved. The levels of reactive oxygen species and malondialdehyde in semen were notably reduced. Hyaluronidase activity and acrosin activity were significantly increased in LBP-treated sperm. However, sperm total motility and DNA integrity were not significantly different between the groups.ConclusionsRSV (50 μM) or LBP (2 mg/mL) are the best candidate antioxidants for inclusion in the freezing medium to improve the quality of giant panda spermatozoa during semen cryopreservation.

Impacts of Social Trust on Rural Households’ Attitudes Towards Ecological Conservation—Example of the Giant Panda Nature Reserves in China

Understanding rural households’ conservation attitudes is crucial to achieving biodiversity conservation effectiveness, and one underlying predictor of household conservation attitudes is social trust. This study examined the impact of rural households’ social trust on their ecological protection attitudes based on 922 rural household data around 13 giant panda nature reserves in Shaanxi Province and Sichuan Province, China. The results show that: (1) Social trust has a significant positive impact on rural households’ conservation attitudes. (2) Males’ ecological conservation attitudes are influenced by all the social trust variables, whereas females’ attitudes are influenced mainly by interpersonal trust (trust in neighbors and villagers). The conservation attitudes of households with higher education levels and higher family incomes are mainly affected by trust in government, while those with lower education levels and lower family incomes are more significantly affected by the trust in villagers and village cadres. The above conclusions are helpful to understand the influencing mechanism of rural households’ conservation attitudes and improve the protection effects of nature reserves.

Single-cell RNA-sequencing analysis and characterisation of testicular cells in giant panda (Ailuropoda melanoleuca).

The giant panda (Ailuropoda melanoleuca ) is a rare and endangered species to be preserved in China. The giant panda has a low reproductive capacity, and due to the scarcity of samples, studies on testes from giant panda are very limited, with little knowledge about the process of spermatogenesis in this species. To establish the gene expression profiles in cells from the testis of a giant panda. The 10×Genomics single-cell RNA-sequencing platform was applied to cells from the testis of an adult giant panda. We identified eight testicular cell types including six somatic and two germ cell types from our single-cell RNA-sequencing datasets. We also identified the differentially expressed genes (DEGs) in each cell type, and performed functional enrichment analysis for the identified testicular cell types. Furthermore, by immunohistochemistry we explored the protein localisation patterns of several marker genes in testes from giant panda. Our study has for the first time established the gene expression profiles in cells from the testis of a giant panda. Our data provide a reference catalogue for spermatogenesis and testicular cells in the giant panda, laying the foundation for future breeding and preservation of this endangered species.

Implications of habitat overlap between giant panda and sambar for sympatric multi-species conservation

Context Studies of distribution and habitat utilisation of sympatric animals, especially those experiencing population increases, are useful for understanding their specialised habitat niches and interspecies relationships, in addition to developing effective protection strategies. Aims As a species that lives within the range of the giant panda, the sambar population has been increasing in the past few decades in Southwest China. In this paper, we aimed to analyse the overlap in habitat area and habitat suitability between giant panda and sambar in Wolong National Nature Reserve to examine potential sympatric species competition in the context of wildlife conservation. Methods We ran MaxEnt models based on giant panda occurrence sign locations (n = 316) and sambar presence locations (n = 598). In addition, we predicted the core and secondary habitat for the two species to assess the degree of competition across a gradient of habitat conditions. Key results Our analysis detected significant habitat overlap between the two species – up to 75.96% in their potential habitat, with suitability overlap indices reaching 0.74. Conclusions These results indicate that the similarity of habitat requirements of the two species is high in our study area. In addition, the competition over space utilisation between the two species’ populations could intensify in the future as the sambar population increases. Implications Our results suggest that habitat restoration and corridor construction could be recommended as conservation strategies for future wildlife conservation in China’s Giant Panda National Park. Moreover, future wildlife conservation should pay greater attention to the niche overlap and interspecific competition among sympatric species. We suggest strengthening habitat restoration and corridors for all key species in the region, not just giant pandas, to alleviate the increased competition associated with niche overlap among sympatric species.

Blood transcriptome analysis revealed the immune changes and immunological adaptation of wildness training giant pandas.

The giant panda (Ailuropoda melanoleuca) is a global flagship species for biodiversity conservation. As the time for captive giant pandas to be released into the wild matures, wildness training is provided to allow adaptation to their natural environment. It is assumed that changes in the immune system would be integral in this adaptation from captive to wild, where many more pathogens would be encountered in their natural habitats. Therefore, this study aims to determine the expression changes of immune-related genes and their potential as immunoassay markers for adaptation monitoring in wildness training giant pandas, and then to understand the adaptation strategy of wildness training giant pandas to the wild environment, thereby improving the success rate of panda reintroduction. We obtained 300 differentially expressed genes (DEGs) by RNA-seq, with 239 up-regulated and 61 down-regulated DEGs in wildness training giant pandas compared to captive pandas. Functional enrichment analysis indicated that up-regulated DEGs were enriched in several immune-related terms and pathways. There were 21 immune-related DEGs, in which most of them were up-regulated in wildness training giant pandas, including several critical innate and cellular immune genes. IL1R2 was the most significantly up-regulated gene and is a signature of homeostasis within the immune system. In the protein-protein interaction (PPI) analysis, CXCL8, CXCL10, and CCL5 were identified as the hub immune genes. Our results suggested that wildness training giant pandas have stronger innate and cellular immunity than captive giant pandas, and we proposed that a gene set of CXCL8, CXCL10, CCL5, CD3D, NFKBIA, TBX21, IL12RB2, and IL1R2 may serve as potential immunoassay markers to monitor and assess the immune status of wildness training giant pandas. Our study offers the first insight into immune alterations of wildness training giant pandas, paving the way for monitoring and evaluating the immune status of giant pandas when reintroducing them into the wild.

Seasonal shift of the gut microbiome synchronizes host peripheral circadian rhythm for physiological adaptation to a low-fat diet in the giant panda.

Characteristics of the gut microbiome vary synchronously with changes in host diet. However, the underlying effects of these fluctuations remain unclear. Here, we performed fecal microbiota transplantation (FMT) of diet-specific feces from an endangered mammal (the giant panda) into a germ-free mouse model. We demonstrated that the butyrate-producing bacterium Clostridium butyricum was more abundant during shoot-eating season than during the leaf-eating season, congruent with the significant increase in host body mass. Following season-specific FMT, the microbiota of the mouse model resembled that of the donor, and mice transplanted with the microbiota from the shoot-eating season grew faster and stored more fat. Mechanistic investigations revealed that butyrate extended the upregulation of hepatic circadian gene Per2, subsequently increasing phospholipid biosynthesis. Validation experiments further confirmed this causal relationship. This study demonstrated that seasonal shifts in the gut microbiome affect growth performance, facilitating a deeper understanding of host-microbe interactions in wild mammals.

Analysis on Change Trend and Influencing Factors of Natural Resource Utilization of Community Farmers around the Habitat of Giant Pandas

Analysis on Change Trend and Influencing Factors of Natural Resource Utilization of Community Farmers around the Habitat of Giant Pandas

Nutritional and Physical Characteristics Evaluation of Giant Panda (Ailuropoda Melanoleuca) Milk, Cow Milk and Goat Milk

Nutritional and Physical Characteristics Evaluation of Giant Panda (Ailuropoda Melanoleuca) Milk, Cow Milk and Goat Milk

The impact of tourist cognition on willing to pay for rare species conservation: Base on the questionnaire survey in protected areas of the Qinling region in China

Wildlife is invaluable for natural development and biodiversity conservation. We investigated tourist cognition of rare species, as well as their economic value, in protected areas of the Qinling region, China. We focused on the tourist cognition and willingness to pay for the conservation of Giant Pandas, Golden Monkeys, Crested Ibises, and Takins. We also explored the influence of the tourist cognition of ecology and its perceived value on the willingness to pay for the conservation of different species. Results indicated that the tourists are more willing to pay for Giant Pandas and Golden Monkeys. The tourist cognition of species affects mainly the willingness to pay, while the tourist cognition of wildlife importance mainly affects the payment. The tourists’ perceived emotional and cost values have significant positive effects on the willingness to pay for the conservation of Giant Pandas/Golden Monkeys and Crested Ibises/Takins, respectively. Education and income levels have significant positive impacts on the willingness to pay and payment, respectively; therefore, it is necessary to publicise rare species, such as Crested Ibis and Takin, through natural education and experience to improve the tourist cognition of species and ecological environment. In addition, innovative forms of product supply combined with the tourists’ needs should be explored to improve their perceived emotional value and willingness to pay for wildlife conservation.