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Abstract Testicular tumours are zoonoses that can occur in not only human, but other animals, include giant pandas. A middle‐aged male giant panda named Fufu was diagnosed with a testicular tumour and underwent surgery to remove the entire left testis. The testis was mainly composed of three substantive parts: normal tissue on the outside, tumour tissue in the middle, and necrosis in the centre. HE stains revealed that the tumour was a seminoma. Single‐cell mRNA sequence was applied to characterise cellular states and molecular circuitries of giant panda testicular seminoma. Only germ cell markers expressed in nearly all tumour cells, and the tumour cells appeared to be the same subtype of seminoma cells. We identified four clusters with unique genes expression. They were early apoptosis cells (EAC), inactive cells (IC), active cells subcluster 1 (AC‐1) and active cells subcluster 2 (AC‐2). We utilised monocle tools and found that IC cells was in the initiation stage, and EAC was one type of terminal stage, suggesting that tumour cells may undergo apoptosis in the future. AC‐2 was another type of terminal stage, representing a group of progressive cells. Our study represents the first report to utilise scRNA‐seq to characterise the cellular states and molecular circuitries of a giant panda testicular tumour. This investigation proposes CD117 and CD30 as dependable markers for future pathologic diagnosis. Our findings also suggest that CTSV and other genes with unique expression patterns in active and progressive giant panda seminoma cells may act as early prognostic biomarkers.

China announced the development of its first 5 national parks in 2021, the primary objective of which is to conserve the natural state and integrity of natural ecosystems. As such, ecosystem services and biodiversity levels are crucial assessment factors for the parks. For Giant Panda National Park (GPNP), we evaluated ecological sensitivity based on water and soil erosion and rocky desertification; ecosystem services based on headwater conservation, soil and water conservation, and biodiversity conservation; and presence of giant panda (Ailuropoda melanoleuca) and sympatric species (e.g., takin [Budorcas taxicolor], Asiatic black bear [Ursus thibetanus]) habitat suitability derived from niche modeling to identify the ecosystem status and assess ecological problems within the park. From our results, we proposed ecologically critical areas to target to meet the park's goals. The suitable habitat for pandas and sympatric species encompassed 62.98% of the park and occurred mainly in the Minshan Mountains. One quarter of the total area (25.67%) contained areas important for ecosystem services. Ecologically sensitive and extremely sensitive areas covered 88.78% of the park and were distributed mainly in Qionglaishan and Minshan Mountains. This coverage indicated that there was much habitat for pandas and sympatric species but that the ecosystems in GPNP are vulnerable. Therefore, ecologically critical areas encompassed all suitable habitats for all the species examined and areas important and extremely important to ecosystem service provision,ecologically sensitive and extremely sensitive areas, encompassed 15.17% of panda habitat, accounted for 16.37% of the GPNP area, and were distributed mainly in the Minshan Mountains. Our results indicated where conservation efforts should be focused in the park and that by identifying ecologically critical areas managers can provide targeted protection for wildlife habitat and ecosystems and effectively and efficiently protect the composite ecosystem. Additionally, our methods can be used to inform development of new national parks. [ABSTRACT FROM AUTHOR]

Abstract As a highly endangered species, the giant panda (panda) has attracted significant attention in the past decades. Considerable efforts have been put on panda conservation and reproduction, offering the promising outcome of maintaining the population size of pandas. To evaluate the effectiveness of conservation and management strategies, recognizing individual pandas is critical. However, it remains a challenging task because the existing methods, such as traditional tracking method, discrimination method based on footprint identification, and molecular biology method, are invasive, inaccurate, expensive, or challenging to perform. The advances of imaging technologies have led to the wide applications of digital images and videos in panda conservation and management, which makes it possible for individual panda recognition in a noninvasive manner by using image‐based panda face recognition method. In recent years, deep learning has achieved great success in the field of computer vision and pattern recognition. For panda face recognition, a fully automatic deep learning algorithm which consists of a sequence of deep neural networks (DNNs) used for panda face detection, segmentation, alignment, and identity prediction is developed in this study. To develop and evaluate the algorithm, the largest panda image dataset containing 6,441 images from 218 different pandas, which is 39.78% of captive pandas in the world, is established. The algorithm achieved 96.27% accuracy in panda recognition and 100% accuracy in detection. This study shows that panda faces can be used for panda recognition. It enables the use of the cameras installed in their habitat for monitoring their population and behavior. This noninvasive approach is much more cost‐effective than the approaches used in the previous panda surveys.

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Simple Summary: The focus of this study was to explore the impact of the transmission of antibiotic resistance genes (ARGs) from a bamboo diet to the gut microbiota of giant pandas and further dissemination through fecal matter. Metagenomic analysis and the Comprehensive Antibiotic Resistance Database (CARD) were used to investigate the presence of ARGs in the gut microbiota of captive giant pandas during the consumption of different parts of bamboo. The results showed that the number of ARGs was highest in gut microbiota of the giant panda during the consumption of bamboo leaves, while the variety of ARGs was highest during the consumption of shoots. Specific bacteria associated with ARG dissemination were also identified. This study emphasizes the need for the proper handling of panda feces and regular monitoring of antimicrobial-resistant genes to mitigate the threat of antibiotic resistance. Overall, this research provides insights into the complex relationship between diet, gut microbiota, and health in giant pandas, contributing to their conservation and management. The spread of antibiotic resistance genes (ARGs) in the environment exacerbates the contamination of these genes; therefore, the role plants play in the transmission of resistance genes in the food chain requires further research. Giant pandas consume different bamboo parts at different times, which provides the possibility of investigating how a single food source can affect the variation in the spread of ARGs. In this study, metagenomic analysis and the Comprehensive Antibiotic Resistance Database (CARD) database were used to annotate ARGs and the differences in gut microbiota ARGs during the consumption of bamboo shoots, leaves, and culms by captive giant pandas. These ARGs were then compared to investigate the impact of bamboo part consumption on the spread of ARGs. The results showed that the number of ARGs in the gut microbiota of the subjects was highest during the consumption of bamboo leaves, while the variety of ARGs was highest during the consumption of shoots. Escherichia coli, which poses a higher risk of ARG dissemination, was significantly higher in the leaf group, while Klebsiella, Enterobacter, and Raoultella were significantly higher in the shoot group. The ARG risk brought by bamboo shoots and leaves may originate from soil and environmental pollution. It is recommended to handle the feces of giant pandas properly and regularly monitor the antimicrobial and virulence genes in their gut microbiota to mitigate the threat of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Simple Summary: Aeromonas veronii, an important aquatic bacterium, has been known to infect a variety of animals worldwide. However, there have been no reports of A. veronii infection in giant pandas. Here, we found that strain VGP was resistant to six antibiotics, carries a large number of resistance genes and virulence genes, and has strong pathogenicity in mice. These results suggest that infection with strain VGP could be one of the leading causes of death for this giant panda cub. Our study presents the first evidence that giant pandas could be infected by A. veronii. The objective of this study was to understand biological characteristics of one bacteria strain named as VPG which was isolated from multiple organs of a dead captive giant panda cub. Here, we use biochemical tests, 16S rRNA and gyrB genes for bacterial identification, the disk diffusion method for antibiotic resistance phenotype, smart chip real-time PCR for the antibiotic resistance genotype, multiplex PCR for determination of virulence genes, and the acute toxicity test in mice for testing the pathogenicity of isolates. The isolate was identified as A. veronii strain based on the biochemical properties and genetic analysis. We found that the strain carried 31 antibiotic resistance genes, revealed antimicrobial resistance phenotypically to several antibiotics including penicillin, ampicillin, oxacillin, amoxicillin, imipenem, and vancomycin, and carried virulence genes including aer, act, lip, exu, ser, luxs, and tapA. The main pathological changes in giant panda were congestion, necrotic lesions and a large number of bacteria in multiple organs. In addition, the LD50 in Kunming mice infected with strain VGP was 5.14 × 107 CFU/mL by intraperitoneal injection. Infection with strain VGP led to considerable histological lesions such as hemorrhage of internal organs, necrosis of lymphocytes and neurons in Kunming mice. Taken together, these results suggest that infection with strain VGP would be an important causes of death in this giant panda cub. [ABSTRACT FROM AUTHOR]

The host range of feline panleukopenia virus (FPV) is expanding and is a serious threat to both captive and free-range endangered wildlife. The FPV named FPV-am2020 was isolated from fecal samples from four diarrheal captive giant pandas in 2020, and pathogenicity and phylogenetic analysis were conducted in this study. Three-month-old cats challenged with FPV-am2020 experienced 100% mortality. The complete FPV-am2020 sequence was determined and comprised 5277 base pairs (bp), 36.76% GC content, and two open reading frames. According to the phylogenetic analysis of whole genome sequences and VP2 gene sequences, FPV-am2020 was closely related with MG764511.1 (isolated from captive lions in China, 2015), KX685354.1 (isolated from captive tigers in China, 2016), and KX900570.1 (isolated from captive jaguar in China, 1986). Furthermore, the study identified a G299E mutation in VP2 which was a key residue involved in phenotype changes in FPV. Thus, increased surveillance of FPV mutant isolates must be enacted to protect giant pandas against potential viral threats. [ABSTRACT FROM AUTHOR]

Simple Summary: Climate change can have a negative impact on the food sources of wild animals, including giant pandas, who primarily rely on bamboo. To better understand the foraging strategy of giant pandas and aid in their conservation, metabolomic methods were used to analyze fecal metabolites and correlate them with gut microbiota. The results indicate that the sugar content in giant panda feces is higher when they consume bamboo culm (the hollow, jointed stem) with high fiber content and that Streptococcus bacteria are positively correlated with glucose and acetic acid content, both of which are related to fiber. These results suggest that the foraging strategy of giant pandas is based on the nutritional composition of different bamboo parts, and therefore, enriching their habitat with bamboo species is essential to achieve their natural foraging strategy to obtain adequate nutrition. Climate change-induced food shortages pose major threats to wildlife conservation, and the exclusive reliance of giant pandas on bamboo makes them particularly vulnerable. The aim of this study was to provide insight into the reasons for the foraging strategies of giant pandas to selectively forage for different bamboo parts (bamboo shoot, culm, and leaf) during different seasons. This study used a metabolomic approach to analyze the fecal metabolites of giant pandas and conducted a correlation analysis with their gut microbiota. The results indicate that the fecal metabolites of giant pandas differ significantly depending on the bamboo parts they forage on, with higher sugar content observed when they consume bamboo culm with high fiber content. By functional annotation, culm group metabolites were enriched in the galactose metabolic pathway, while shoot group metabolites were enriched in the phenylalanine, tyrosine and tryptophan biosynthesis pathways. Moreover, Streptococcus showed a significant positive correlation with glucose and acetic acid content. Therefore, the foraging strategy of giant pandas is based on the ability to utilize the nutrient content of different bamboo parts. Captive feeding and habitat construction should enrich bamboo species to allow them to express their natural foraging strategies and improve their welfare and reproductive status. [ABSTRACT FROM AUTHOR]

The giant panda (Ailuropoda melanoleuca) and the Chinese red panda (Ailurus styani) are distributed in the same region in the mountain forest ecosystem on the eastern edge of the Qinghai Tibet Plateau and share the same food sources. In order to understand how sympatric giant pandas and Chinese red pandas maintain interspecific relationships to achieve stable coexistence, we used species distribution models and diurnal activity rhythms to analyze the spatial and temporal niche characteristics of giant pandas and Chinese red pandas in the Daxiangling Mountain system based on 187 camera traps data. The results show that: (1) In the Daxiangling Mountains, the total area of suitable habitats for giant pandas and Chinese red pandas is 717.61 km2 and 730.00 km2, respectively, accounting for 17.78% and 18.25%, respectively, of the study area. (2) The top five environmental factors contributing to the model of giant panda and Chinese red panda are precipitation seasonality, temperature seasonality, distance to the road, and elevation and vegetation type. (3) The total overlapping area of suitable habitats for giant pandas and Chinese red pandas is 342.23 km2, of which the overlapping area of highly suitable habitats is 98.91 km2. The overlapping index of suitable habitats is 0.472, and the overlapping index of highly suitable habitats is 0.348, which indicates that the two achieve spatial niches are separated to achieve stable coexistence. (4) The overlapping index of the daily activity rhythm of giant panda and Chinese red panda is 0.87, which is significantly different (p <.05). The existence of Chinese red panda will significantly affect the daily activity rhythm of giant panda (p <.001). This research can provide scientific reference for the researches about population and habitat protection of giant pandas and Chinese red pandas, so as to understand the driving mechanism of resource allocation and population dynamics of sympatric species. [ABSTRACT FROM AUTHOR]

The conservation of the giant panda (Ailuropoda melanoleuca), as an iconic vulnerable species, has received great attention in the past few decades. As an important part of the giant panda population survey, the age distribution of giant pandas can not only provide useful instruction but also verify the effectiveness of conservation measures. The current methods for determining the age groups of giant pandas are mainly based on the size and length of giant panda feces and the bite value of intact bamboo in the feces, or in the case of a skeleton, through the wear of molars and the growth line of teeth. These methods have certain flaws that limit their applications. In this study, we developed a deep learning method to study age group classification based on facial images of captive giant pandas and achieved an accuracy of 85.99% on EfficientNet. The experimental results show that the faces of giant pandas contain some age information, which mainly concentrated between the eyes of giant pandas. In addition, the results also indicate that it is feasible to identify the age groups of giant pandas through the analysis of facial images. [ABSTRACT FROM AUTHOR]

The giant panda (Ailuropoda melanoleuca) has long attracted the attention of conservationists as a flagship and umbrella species. Collecting attribute information on the age structure and sex ratio of the wild giant panda populations can support our understanding of their status and the design of more effective conservation schemes. In view of the shortcomings of traditional methods, which cannot automatically recognize the age and sex of giant pandas, we designed a SENet (Squeeze-and-Excitation Network)-based model to automatically recognize the attributes of giant pandas from their vocalizations. We focused on the recognition of age groups (juvenile and adult) and sex of giant pandas. The reason for using vocalizations is that among the modes of animal communication, sound has the advantages of long transmission distances, strong penetrating power, and rich information. We collected a dataset of calls from 28 captive giant panda individuals, with a total duration of 1298.02 s of recordings. We used MFCC (Mel-frequency Cepstral Coefficients), which is an acoustic feature, as inputs for the SENet. Considering that small datasets are not conducive to convergence in the training process, we increased the size of the training data via SpecAugment. In addition, we used focal loss to reduce the impact of data imbalance. Our results showed that the F1 scores of our method for recognizing age group and sex reached 96.46% ± 5.71% and 85.85% ± 7.99%, respectively, demonstrating that the automatic recognition of giant panda attributes based on their vocalizations is feasible and effective. This more convenient, quick, timesaving, and laborsaving attribute recognition method can be used in the investigation of wild giant pandas in the future. [ABSTRACT FROM AUTHOR]

Simple Summary: In the process of ex-situ conservation, young giant pandas face a variety of unfavorable environmental impacts such as frequent maternal separation, parenting by non-parents, noise from tourists, and frequent replacement of animal houses, which may cause psychological and physiological stress. The gut microbiota is an important carrier of the interaction between the body and the environment, and recent studies revealed an association between stress and alterations of the intestinal microbiota. So, is the stress caused by the unfavorable parenting environment in the early life of captive giant pandas related to the gut microbiota? To answer this question, we use fecal metagenomics and LC-MS technology to study the effect of different parenting patterns on the structure, diversity, and metabolites of the intestinal microbial community of captive giant pandas. In order to evaluate the possible adverse effects of the traditional parenting mode on the gut microbiota of captive giant pandas in the early life, it can provide an important scientific basis for improving the welfare level of captive giant pandas. Maternal deprivation (MD) in early life induces dysbiosis in the host gut microbiota, which is a key determinant of abnormal behavior in stress model individuals. Compared with the early parenting environment of the wild, captive giant pandas face frequent and premature maternal separation. Will this lead to imbalance in intestinal flora and stress in captive giant pandas? The purpose of this research is to evaluate the possible adverse effects of the traditional parenting mode on the gut microbiota of captive giant pandas. The results showed that the frequent and premature maternal separation at early stages of the young did not change α and β diversity indices of the gut microbes, but it increased the relative abundance of s_Clostridium_tetani and s_Clostridium sp MSJ_8 (significantly positively correlated with the metabolism of propionic acid) and also the concentrations of fecal metabolites that are related to stress (N-acetyl-l-aspartic acid and corticosterone) in the intestinal tract of giant pandas in adulthood. Thereby, the function of protein digestion and absorption in the intestines of captive giant pandas was decreased, and the metabolism of short-chain fatty acids was disturbed. In conclusion, the parenting experience of early maternal separation could adversely affect the stress caused by the unfavorable parenting environment in the early life of captive giant pandas related to the gut microbiota of the captive giant pandas in adulthood. [ABSTRACT FROM AUTHOR]

Context: 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. Aims: To establish the gene expression profiles in cells from the testis of a giant panda. Methods: The 10× Genomics single-cell RNA-sequencing platform was applied to cells from the testis of an adult giant panda. Key results: 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. Conclusions: Our study has for the first time established the gene expression profiles in cells from the testis of a giant panda. Implications: 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. The giant panda (Ailuropoda melanoleuca) is a rare and endangered species to be preserved in China. Here, we employed the 10× Genomics single-cell RNA-sequencing platform to establish the gene expression profiles in cells from the testis of an adult giant panda. Hence, this study provides a reference catalogue for spermatogenesis and testicular cells in giant panda, laying the foundation for future research on breeding, preservation, diagnosis, and treatment of reproductive disorders in this endangered species. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is a worldwide-distributed zoonotic protozoan parasite which causes toxoplasmosis and has a significant effect on public health. In the giant panda (Ailuropoda melanoleuca), toxoplasmosis can cause asymptomatic infections, reproductive disorder and even death, which poses a serious threat to the conservation of this rare protected species. Therefore, serological investigation of T. gondii is essential to understanding its risk to giant pandas, however, there are no specific testing kits for giant pandas. Previous research has used MAT as the reference method for screening T. gondii , to investigate this further, this study focused on the agreement comparing of MAT with ELISA and IHA tests for detecting T. gondii antibodies in 100 blood samples from 55 captive giant pandas in Chengdu, China. The results showed 87.0%, 87.0%, 84.0%, samples were sero-positive for T. gondii using ELISA (kits a, b, c), respectively, while MAT and IHA tests were 84.0% and 9.0% sero-positive, respectively. There was no significant difference between MAT and the three ELISA kits and these two methods had substantial agreement (0.61 < қ ≤ 0.80). Meanwhile, there was a significant difference (P < 0.001) between MAT and IHA, and these two methods had only a slight agreement (қ ≤ 0.20). The relative sensitivity of the ELISA (kits a, b, c) were 89.0%, 91.5% and 95.1%, and the specificity were 86.7%, 80.0% and 80.0%, respectively, which showed these three ELISA kits all had great accuracy. It is suggested that MAT is the recommended test method for primary screening T. gondii in giant pandas and then verified by ELISA. [Display omitted] • It's the first report to compare the agreement of the three test kits for detecting T. gondii antibodies in giant panda. • No significant difference and substantial agreement between MAT and ELISA, slight agreement, meanwhile, the contrary result between MAT and IHA. • The relative sensitivity of ELISA kits was 89%, 91.5% and 95.1%, and the specificity was 86.7%, 80% and 80%, respectively. • The two commonly used commercial MAT and ELISA kits tested were valuable tools for T. gondii diagnosis for giant panda. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Dental caries severely hinders efficient access to adequate energy in wildlife. Different food supplies will develop characteristic plaque, and the microorganisms of these plaque are closely related to dental health. Here, plaque samples from panda cubs with caries and caries-free were collected for 16S rRNA high-throughput sequencing. All sequences clustered into 337 operational taxonomic units (OTUs; 97% identity), representing 268 independent species belonging to 189 genera, 98 families, 51 orders, 24 classes, and 13 phyla. Two groups shared 218 OTUs, indicating the presence of a core plaque microbiome. α diversity analysis showed that the microbial diversity in plaques with caries exceeded that of caries-free. The dominant phyla of plaque microbiota included Proteobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Actinobacteria. The dominant genera included unclassified Neisseriaceae , Actinobacillus , Lautropia , Neisseria , Porhyromonas , unclassified Pasteurellaceae , Moraxella , Streptococcus , Bergeywlla and Capnocytophaga. β diversity analysis showed that the plaque microbial community structure was different between two groups. Using LEfSe analysis, 19 differentially abundant taxa were identified as potential biomarkers. Finally, function predictions analysis showed All the energy related metabolic pathways on KEGG level 2 were enriched in caries-active group. Consistent with the mainstream caries-causing narrative, our results illuminate the lack of information regarding the oral microflora composition and function within giant panda cubs. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae is not only a worldwide human pathogen, it also effects wildlife, such as the giant panda (Ailuropoda melanoleuca), in which it has recently been evidenced to result in diarrhea, organ failure, and even death. A K. pneumoniae investigation was carried out at the Chengdu Research Base of Giant Panda Breeding in 2018. As part of the investigation, the pulsed-field gel electrophoresis (PFGE) typing, multilocus-sequence typing (MLST), antibiotic resistance profiles (ARPs), and antibiotic resistance genes (ARGs) were studied based on all isolates. Fecal samples were collected from 72 A. melanoleuca from May to December 2018, and a total of 90 K. pneumoniae were isolated from 153 fecal samples. The genotyping results showed that the isolates had high diversity, of which 84 clusters were obtained by PFGE and 57 STs by MLST. The overall trend of the similarity of isolates was the first sample period > second sample period > third sample period, which showed the increasement of genome variability of K. pneumoniae. In addition, 90 isolates showed high resistance to ampicillin, rifampicin, and compound sulfamethoxazole. Of the obtained isolates, 50% carried 6~8 ARPs, and the carrying volume increased during three sample periods, in which we found two isolates carrying 12 and 13 ARPs during the third sample period, respectively. Moreover, a total of 65 ARGs were detected (90.28%, 65/72) in 90 K. pneumoniae samples. Almost all bacteria sampled contained 17 ARGs that belonged to the β-lactamase, Multidrug, MGEs, Aminoglycoside, and Tetracycline, which may be the basis of ARPs of K. pneumoniae. Moreover, the types of Multidrug and MGEs had a greater impact on antibiotic susceptivity of K. pneumoniae. Our results showed that K. pneumoniae has a serious risk of transmission in A. melanoleuca and K. pneumoniae had a high possibility of genome diversity and the risk of drugs tolerance under the large antibiotic usage. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Knowledge of energy expenditure informs conservation managers for long term plans for endangered species health and habitat suitability. We measured field metabolic rate (FMR) of free-roaming giant pandas in large enclosures in a nature reserve using the doubly labeled water method. Giant pandas in zoo like enclosures had a similar FMR (14,182 kJ/day) to giant pandas in larger field enclosures (13,280 kJ/day). In winter, giant pandas raised their metabolic rates when living at − 2.4 °C (36,108 kJ/day) indicating that they were below their thermal neutral zone. The lower critical temperature for thermoregulation was about 8.0 °C and the upper critical temperature was about 28 °C. Giant panda FMRs were somewhat lower than active metabolic rates of sloth bears, lower than FMRs of grizzly bears and polar bears and 69 and 81% of predicted values based on a regression of FMR versus body mass of mammals. That is probably due to their lower levels of activity since other bears actively forage for food over a larger home range and pandas often sit in a patch of bamboo and eat bamboo for hours at a time. The low metabolic rates of giant pandas in summer, their inability to acquire fat stores to hibernate in winter, and their ability to raise their metabolic rate to thermoregulate in winter are energetic adaptations related to eating a diet composed almost exclusively of bamboo. Differences in FMR of giant pandas between our study and previous studies (one similar and one lower) appear to be due to differences in activity of the giant pandas in those studies. [ABSTRACT FROM AUTHOR]

Background: Flavonoids are important plant secondary metabolites (PSMs) that have been widely used for their health-promoting effects. However, little is known about overall flavonoid metabolism and the interactive effects between flavonoids and the gut microbiota. The flavonoid-rich bamboo and the giant panda provide an ideal system to bridge this gap. Results: Here, integrating metabolomic and metagenomic approaches, and in vitro culture experiment, we identified 97 flavonoids in bamboo and most of them have not been identified previously; the utilization of more than 70% flavonoid monomers was attributed to gut microbiota; the variation of flavonoid in bamboo leaves and shoots shaped the seasonal microbial fluctuation. The greater the flavonoid content in the diet was, the lower microbial diversity and virulence factor, but the more cellulose-degrading species. Conclusions: Our study shows an unprecedented landscape of beneficial PSMs in a non-model mammal and reveals that PSMs remodel the gut microbiota conferring host adaptation to diet transition in an ecological context, providing a novel insight into host-microbe interaction. FNqEF651ov9tHyt7pvrVsG Video abstract [ABSTRACT FROM AUTHOR]

Bacterial infection and imbalance of bacterial community in the genitourinary system of giant panda could affect the reproductive health. In severe cases, it can also lead to abortion. In this study, 13 of vaginal secretions in the estrue (E) group and seven of vaginal secretions in the non-estrue (NE) group were used to study the composition and diversity of vaginal bacterial communities between estrus and non-estrus by 16S rRNA gene sequencing analysis. The results showed that the vaginal microbiome in giant pandas shared the same top five abundant species between estrus and non-estrus at the phylum level. However, the vaginal microbiome changed significantly during estrus at the genus level. In top 10 genera, the abundance of Escherichia , Streptococcus , and Bacteroides in the E group was significantly higher than that in the NE group (p <0.05); Azomonas , Porphyromonas , Prevotella , Campylobacter , and Peptoniphilus in the NE group was significantly higher than that in the E group (p <0.05). The richness and diversity of vaginal microbiome in giant panda on estrus were significantly lower than those on non-estrus (p <0.05). It is noteworthy that the abundance of Streptococcus , Escherichia , and Bacteroides of vagina in giant pandas maintained low abundance in the daily. Whereas, they increased significantly during estrus period, which may play an important role in female giant pandas during estrus period. It was hypothesized that hormones may be responsible for the changes in the vaginal microbiome of giant pandas between estrus and no-estrus stages. [ABSTRACT FROM AUTHOR]

Giant pandas consume different structural parts of bamboo (shoots, leaves and culms) during different seasons. Previous research showed different bamboo parts have varying nutritional content and that a long-term diet consisting of a single part of bamboo resulted in remarkable metabolic changes within captive giant pandas. However, the effects on the gut microbiome of giant pandas, as a result of a single bamboo part diet, have not been investigated. Here, we evaluated the changes in gut microbial communities based on single bamboo part diets and their potential implications by using 16S rRNA gene-based amplicon sequencing and metagenome shotgun sequencing. We found that the composition and function of the gut microbiome from captive giant pandas fed exclusively culms were significantly different from that of individuals fed shoots or leaves. During the culm feeding period, the gut microbiome showed strongest digestive capabilities for cellulose, hemicellulose and starch, and had the highest potential abilities for the biosynthesis of bile acids, fatty acids and amino acids. This suggests the microbiome aids in breaking down culm, which is more difficult for giant pandas to digest, as a means to compensate for the nutrient poor content of the culm. Genes related to fatty acid metabolism and tricarboxylic acid cycle enzymes were more abundant during the leaf stage diet than that in the shoot and culm stages. Thus, the microbiome may help giant pandas, which typically have low lipase levels, with fat digestion. These results illustrate that adaptive changes in the gut microbiome community and function may be an important mechanism to aid giant panda digestion when consuming different structural parts of bamboo. [ABSTRACT FROM AUTHOR]

Canine distemper (CD) is a significant threat to wild and captive giant panda populations. Captive giant pandas are inoculated with canine distemper virus (CDV) vaccination to prevent the infection with the CDV. As an important regulator, microRNA (miRNA) plays a crucial role in regulating gene expression, including in disease immunity. To understand the role of miRNA in immune response to CDV vaccination, we investigated the miRNA expression profile in five giant panda cubs after two inoculations, 21 days apart. A total of 187 conserved miRNAs and 96 novel miRNAs were identified. Among the 187 conserved miRNAs, 29 differentially expressed miRNAs were found postinoculation. The upregulation of miR-16, miR-182, miR-30b, and miR-101 indicated that the innate immune may be enhanced, whereas the upregulation of miR-142 and miR-19a are probably involved in the enhanced cellular immune response. However, the downregulated miR-155 and miR-181a might indicate the giant panda has weak ability to produce antibodies and memory B cells. Integrated analysis of miRNA-messenger RNA (mRNA) found 20 negatively regulated miRNA-mRNA pairs, where downregulated miR-204 might enhance giant panda cub innate immunity by increasing TLR6 expression, and downregulated miR-330 might activate macrophages and regulate the immune response by increasing TMEM106A expression. Our research provides key information for future development to enhance the immune response of giant pandas and potentially improve the survival of captive and wild giant panda populations threatened by CD. [ABSTRACT FROM AUTHOR]

Giant pandas (Ailuropoda melanoleuca) were historically hunted using dogs and are currently threatened by free-roaming dogs and their associated diseases. To better understand the spatial magnitude of this threat, we used a GIS approach to investigate edge effects of dogs on giant panda habitat. We first examined two nature reserves with contrasting free-roaming dog populations: Liziping, with many dogs (~0.44/km2), and Daxiangling, with few dogs (~0.14/km2). Spatial analysis indicated that giant pandas at Liziping (but not Daxiangling) showed a shift in habitat use away from populated areas consistent with a risk response to the foray distance of free-roaming dogs (10.9 km path-distance). Most giant panda locations (86%) from the 2014 census in Liziping were clustered around remote "dog-free zones." Expanding this analysis across the entire giant panda range revealed that 40% of panda habitat is within the foray distance of dogs. Our assessment will inform dog control programs including monitoring, education, veterinary care, and other measures. We recommend that reserves designated for the release of translocated pandas receive priority consideration for dog control efforts. Only by understanding and managing complex interactions between humans, domestic animals, and wild animals can we sustain natural systems in a world increasingly dominated by humans. [ABSTRACT FROM AUTHOR]

Giant pandas represent one of the most endangered species worldwide, and their reproductive capacity is extremely low. They have a relatively long gestational period, mainly because embryo implantation is delayed. Giant panda cubs comprise only a small proportion of the mother's body weight, making it difficult to determine whether a giant panda is pregnant. Timely determination of pregnancy contributes to the efficient breeding and management of giant pandas. Meanwhile, metabolomics studies the metabolic composition of biological samples, which can reflect metabolic functions in cells, tissues, and organisms. This work explored the urinary metabolites of giant pandas during pregnancy. A sample of 8 female pandas was selected. Differences in metabolite levels in giant panda urine samples were analyzed via ultra-high-performance liquid chromatography/mass spectrometry comparing pregnancy to anoestrus. Pattern recognition techniques, including partial least squares-discriminant analysis and orthogonal partial least squares-discriminant analysis, were used to analyze multiple parameters of the data. Compared with the results during anoestrus, multivariate statistical analysis of results obtained from the same pandas being pregnant identified 16 differential metabolites in the positive-ion mode and 43 differential metabolites in the negative-ion mode. The levels of tryptophan, choline, kynurenic acid, uric acid, indole-3-acetaldehyde, taurine, and betaine were higher in samples during pregnancy, whereas those of xanthurenic acid and S-adenosylhomocysteine were lower. Amino acid metabolism, lipid metabolism, and organic acid production differed significantly between anoestrus and pregnancy. Our results provide new insights into metabolic changes in the urine of giant pandas during pregnancy, and the differential levels of metabolites in urine provide a basis for determining pregnancy in giant pandas. Understanding these metabolic changes could be helpful for managing pregnant pandas to provide proper nutrients to their fetuses. [ABSTRACT FROM AUTHOR]

The potential threat of domestic dogs to wildlife habitat in China is not widely recognized, despite their large population, lack of regulations regarding their control, and threat they pose to native species. In a case study in 2017, we surveyed villages surrounding Liziping Nature Reserve, the primary site for the release of captive‐born giant pandas (Ailuropoda melanoleuca) into the wild. We conducted surveys of dog owners to assess the population size, demographics, free‐roaming status, and vaccination and sterilization history of their dogs. We collected blood and fecal samples to assess the prevalence of viral and parasite disease threats. At least 370 owned dogs lived near the core giant panda habitat; 64% were free‐roaming, 21% had positive antibody titers for ≥1 of the 4 viruses we tested (canine distemper, parvovirus, rotavirus, rabies), and 67% were positive for gastrointestinal parasites. The high proportion of free‐roaming dogs, uninhibited access to the reserve, and high prevalence of infectious diseases indicate that dogs pose a serious threat to wildlife within Liziping. The extent of this threat throughout the giant panda nature reserve network is unknown and should be assessed. © 2019 The Authors. Journal of Wildlife Management published by Wiley Periodicals, Inc. on behalf of The Wildlife Society. Numerous free‐ranging domestic dogs were located close to Liziping National Nature Reserve, a primary location of giant panda reintroductions and conservation. Viral and parasitic disease threats to giant pandas and other wildlife were present in the population, indicating a need for immediate management action. [ABSTRACT FROM AUTHOR]

Background/Aims: Giant pandas, an endangered species, are a powerful symbol of species conservation. Giant pandas may suffer from a variety of diseases. Owing to their highly specialized diet of bamboo, giant pandas are thought to have a relatively weak ability to resist diseases. The spleen is the largest organ in the lymphatic system. However, there is little known about giant panda spleen at a molecular level. Thus, clarifying the regulatory mechanisms of spleen could help us further understand the immune system of the giant panda as well as its conservation. Methods: The two giant panda spleens were from two male individuals, one newborn and one an adult, in a non-pathological condition. The whole transcriptomes of mRNA, lncRNA, miRNA, and circRNA in the two spleens were sequenced using the Illumina HiSeq platform. EBseq and IDEG6 were used to observe the differentially expressed genes (DEGs) between these two spleens. Gene Ontology and KEGG analyses were used to annotate the function of DEGs. Furthermore, networks between non-coding RNAs and protein-coding genes were constructed to investigate the relationship between non-coding RNAs and immune-associated genes. Results: By comparative analysis of the whole transcriptomes of these two spleens, we found that one of the major roles of lncRNAs could be involved in the regulation of immune responses of giant panda spleens. In addition, our results also revealed that microRNAs and circRNAs may have evolved to regulate a large set of biological processes of giant panda spleens, and circRNAs may function as miRNA sponges. Conclusion: To our knowledge, this is the first report of lncRNAs and circRNAs in giant panda, which could be a useful resource for further giant panda research. Our study reveals the potential functional roles of miRNAs, lncRNAs, and circRNAs in giant panda spleen. [ABSTRACT FROM AUTHOR]

To forecast giant panda (Ailuropoda melanoleuca) population dynamics in the wild, it is crucial to comprehend their age distribution. Traditional methods for estimating the age of panda are costly, time-consuming, and inaccurate. Additionally, these methods only forecast an age group rather than a real age, and lack a uniform standard. However, advances in deep learning and computer vision have given rise to fresh approaches to this problem. Classification models can be improved by using ordinal regression, which uses ordinal correlations across ages to reduce the non-stationary nature of aging tasks. In this study, we collected 8002 images from 272 pandas in various environments, whose ages ranged from 0 to 38. We applied a five-fold subject-exclusive (SE) protocol to train seven Convolutional Neural Networks (CNN) based on ordinal regression. Experiments were conducted on the Panda Age Dataset (PAD Full) and the Lite Panda Age Dataset (PAD Lite). The results were very encouraging and achieved a Mean Absolute Error (MAE) of 2.51 and 2.41, respectively. Our findings demonstrate that this new tool can noninvasively predict the age of giant pandas in captivity and the wild. Continued development of computer vision technology will drive progress in ecology and conservation. • Machine learning can be used to predict the age of giant pandas based on facial images. • Ordinal regression can enhance model performance based on age information. • The inference speed of model is as important as its accuracy. [ABSTRACT FROM AUTHOR]

A novel bacterial strain, designated as CF21, was isolated from the air of Ailuropoda melanoleuca enclosures in China. Cells were gram-negative, aerobic, non-motile, and rod shaped. Strain CF21 grew at 10-40 °C (optimum 28-30 °C) and pH 6.0-9.0 (optimum pH 7.0-8.0) and in the presence of NaCl concentrations ranging from 0.0 % (w/v) to 2.0 % (optimum 0.0-1.0 %). 16SrRNA gene sequence analysis indicated that strain CF21 belonged to genus Lysobacter within class Gammaproteobacteria and was most closely related to Luteimonas dalianensi OB44-3 (95.8 % similarity), Lysobacter ruishenii CTN-1 (95.1 %), Lysobacter spongiicola KMM329 (94.8 %), and Lysobacter daejeonensis GH1-9 (94.6 %). The genomic G+C DNA content was 68.72 mol%. Major cellular fatty acids of CF21 were iso-C (30.22 %), iso-C (25.70 %), and the sum of 10-methyl C and/or iso-Cω9c (21.94 %). The prominent isoprenoid quinone was ubiquinone 8 (Q-8). Primary polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unknown phospholipid. DNA sequence relatedness between strain CF21 and L. ruishenii CTN-1 was 56 %, which was clearly below the 70 % threshold for prokaryotic species delineation. These analyses indicated that CF21 is a novel member of genus Lysobacter, for which the name Lysobacter chengduensis sp. nov. is proposed. The type strain is CF21 (=CGMCC1.15145 = DSM 100306). [ABSTRACT FROM AUTHOR]

Ursids (bears) in general, and giant pandas in particular, are highly altricial at birth. The components of bear milks and their changes with time may be uniquely adapted to nourish relatively immature neonates, protect them from pathogens, and support the maturation of neonatal digestive physiology. Serial milk samples collected from three giant pandas in early lactation were subjected to untargeted metabolite profiling and multivariate analysis. Changes in milk metabolites with time after birth were analysed by Principal Component Analysis, Hierarchical Cluster Analysis and further supported by Orthogonal Partial Least Square-Discriminant Analysis, revealing three phases of milk maturation: days 1–6 (Phase 1), days 7–20 (Phase 2), and beyond day 20 (Phase 3). While the compositions of Phase 1 milks were essentially indistinguishable among individuals, divergences emerged during the second week of lactation. OPLS regression analysis positioned against the growth rate of one cub tentatively inferred a correlation with changes in the abundance of a trisaccharide, isoglobotriose, previously observed to be a major oligosaccharide in ursid milks. Three artificial milk formulae used to feed giant panda cubs were also analysed, and were found to differ markedly in component content from natural panda milk. These findings have implications for the dependence of the ontogeny of all species of bears, and potentially other members of the Carnivora and beyond, on the complexity and sequential changes in maternal provision of micrometabolites in the immediate period after birth. [ABSTRACT FROM AUTHOR]

The giant panda (Ailuropoda melanoleuca) is one of the world’s most beloved endangered mammals. Although the draft genome of this species had been assembled, little was known about the composition of its microRNAs (miRNAs) or their functional profiles. Recent studies demonstrated that changes in the expression of miRNAs are associated with immunity. In this study, miRNAs were extracted from the blood of four healthy giant pandas and sequenced by Illumina next generation sequencing technology. As determined by miRNA screening, a total of 276 conserved miRNAs and 51 novel putative miRNAs candidates were detected. After differential expression analysis, we noticed that the expressions of 7 miRNAs were significantly up-regulated in young giant pandas compared with that of adults. Moreover, 2 miRNAs were up-regulated in female giant pandas and 1 in the male individuals. Target gene prediction suggested that the miRNAs of giant panda might be relevant to the expressions of 4,602 downstream genes. Subseuqently, the predicted target genes were conducted to KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis and we found that these genes were mainly involved in host immunity, including the Ras signaling pathway, the PI3K-Akt signaling pathway, and the MAPK signaling pathway. In conclusion, our results provide the first miRNA profiles of giant panda blood, and the predicted functional analyses may open an avenue for further study of giant panda immunity. [ABSTRACT FROM AUTHOR]

It has been widely known that the giant panda (Ailuropoda melanoleuca) is one of the most endangered species in the world. An optimized platform for maintaining the proliferation of giant panda mesenchymal stem cells (MSCs) is very necessary for current giant panda protection strategies. Basic fibroblast growth factor (bFGF), a member of the FGF family, is widely considered as a growth factor and differentiation inducer within the stem cell research field. However, the role of bFGF on promoting the proliferation of MSCs derived from giant panda bone marrow (BM) has not been reported. In this study, we aimed to investigate the role of bFGF on the proliferation of BM-MSCs derived from giant panda. MSCs were cultured for cell proliferation analysis at 24, 48 and 72 hrs following the addition of bFGF. With increasing concentrations of bFGF, cell numbers gradually increased. This was further demonstrated by performing 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) cell proliferation assay, 5-Bromo-2-deoxyUridine (BrdU) labeling and cell cycle testing. Furthermore, the percentage of MSCs that were OCT4 positive increased slightly following treatment with 5 ng/ml bFGF. Moreover, we demonstrated that the extracellular signal-regulated kinase (ERK) signaling pathway may play an important role in the proliferation of panda MSCs stimulated by bFGF. In conclusion, this study suggests that giant panda BM-MSCs have a high proliferative capacity with the addition of 5 ng/ml bFGF in vitro. [ABSTRACT FROM AUTHOR]

The giant panda ( Ailuropoda melanoleuca) is one of the most famous flagship species for conservation, and its draft genome has recently been assembled. However, the transcriptome is not yet available. In this study, the blood transcriptomes of three pandas were characterized and about 160 million sequencing reads were generated using Illumina Hi Seq 2000 paired-end sequencing technology. The assembly yielded 92 598 transcripts with an average length of 1626 bp and N50 length of 2842 bp. Based on a sequence similarity search against nonredundant (nr) protein database, a total of 38 522 (41.6%) transcripts were annotated. Of these annotated transcripts, 25 142 and 8272 transcripts were assigned to gene ontology terms and clusters of orthologous group, respectively. A search against the Kyoto Encyclopedia of Genes and Genomes Pathway database ( KEGG) indicated that 9098 (9.83%) transcripts mapped to 324 KEGG pathways, and the best represented functional categories of pathways were signal transduction and immune system. We have also identified 23 460 microsatellites, 43 560 SNPs as well as 21 456 alternative splicing events in the assembly. Additionally, a total of 24 341 complete open reading frames ( ORFs) were detected from the assembly where 1492 ORFs were found to be novel gene loci as these have not been annotated so far in any public database. [ABSTRACT FROM AUTHOR]

Males will alter their mating behavior to cope with the presence of their competitors. Even exposure to odors from potential competitors can greatly increase male ejaculate expenditure in a variety of animals including insects, fishes, birds and rodents. Major efforts have been made to examine males' plastic responses to sperm competition and its fitness benefits. However, the effects of competitor absence on male's sexual motivation and behaviors remain unclear, which has been proposed to be one of the causes for the poor sexual performance of some captive mammals. This study revealed that sexual motivation can be greatly enhanced in captive male giant pandas (Ailuropoda melanoleuca) by exposure to chemosensory cues from either one or three conspecifics males. It had been shown that potential rivals' odors increased males' chemosensory investigation behavior, as well as their observing, following and sniffing behaviors towards estrous females. Behaviors changed regardless of the number of rivals (one or three). Our results demonstrate the effects of potential competition on male giant pandas' sexual motivation and behavioral coping strategy. We anticipate that our research will provide a fresh insight into the mechanisms underlying poor sexual performance in male captive mammals, and valuable information for the practical management and ex situ conservation of endangered species. [ABSTRACT FROM AUTHOR]

Recent success in breeding giant pandas in captivity has encouraged panda conservationists to believe that the ex situ population is ready to serve as a source for supporting the wild population. In this study, we used 11 microsatellite DNA markers to assess the amount and distribution of genetic variability present in the two largest captive populations (Chengdu Research Base of Giant Panda Breeding, Sichuan Province and the China Research and Conservation Center for the Giant Panda at Wolong, Sichuan Province). The data were compared with those samples from wild pandas living in two key giant panda nature reserves (Baoxing Nature Reserve and Wanglang Nature Reserve). The results show that the captive populations have retained lower levels of allelic diversity and heterozygosity compared to isolated wild populations. However, low inbreeding coefficients indicate that captive populations are under careful genetic management. Excessive heterozygosity suggests that the two captive populations have experienced a genetic bottleneck, presumably caused by founder effects. Moreover, evidence of increased genetic divergence demonstrates restricted breeding options within facilities. Based on these results, we conclude that the genetic diversity in the captive populations is not optimal. Introduction of genetic materials from wild pandas and improved exchange of genetic materials among institutions will be necessary for the captive pandas to be representative of the wild populations. [ABSTRACT FROM AUTHOR]

cDNA encoding pituitary (PRL) of giant panda was obtained using RT-PCR and expressed in E. coli . The results revealed that panda PRL cDNA encodes a precursor protein of 229 amino acids including a putative signal peptide of 30 amino acids and a mature protein of 199 residues with one potential N-glycosylation site. Sequence comparison indicated that panda PRL shares a high degree of identity to other known PRL sequences ranging from 98% with mink PRL to about 50% with rodent PRL. Six cysteine residues and 29 conserved residues distributed in four domains (PD1, PD2, PD3, and PD4) of PRL were observed through multiple sequence alignment. Fourteen key residues of binding sites 1 and 2 involved in receptor binding are conserved in panda PRL. GST fused recombinant panda PRL protein was efficiently expressed with the form of insoluble inclusion bodies in E. coli BL21 transformed with a pGEX-4T-1 expression vector containing the DNA sequence encoding mature panda PRL. Western blot analysis indicated that GST-panda PRL recombinant protein could be recognized by antibody against human PRL. Our results would contribute to further elucidating the structural and functional characteristics of pituitary PRL and provide a basis for the production of recombinant panda prolactin for future use in the breeding of giant panda. [ABSTRACT FROM AUTHOR]