China Conservation and Research Center for Giant Panda

Wolong, China

China Conservation and Research Center for Giant Panda

Wolong, China
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Ling S.S.,China Conservation and Research Center for Giant Panda | Zhu Y.,Sichuan Agricultural University | Lan D.,Sichuan Agricultural University | Li D.S.,China Conservation and Research Center for Giant Panda | And 7 more authors.
Genetics and Molecular Research | Year: 2017

The giant panda, Ailuropoda melanoleuca (Ursidae), has a unique bamboo-based diet; however, this low-energy intake has been sufficient to maintain the metabolic processes of this species since the fourth ice age. As mitochondria are the main sites for energy metabolism in animals, the protein-coding genes involved in mitochondrial respiratory chains, particularly cytochrome c oxidase subunit II (COX2), which is the rate-limiting enzyme in electron transfer, could play an important role in giant panda metabolism. Therefore, the present study aimed to isolate, sequence, and analyze the COX2 DNA from individuals kept at the Giant Panda Protection and Research Center, China, and compare these sequences with those of the other Ursidae family members. Multiple sequence alignment showed that the COX2 gene had three point mutations that defined three haplotypes, with 60% of the sequences corresponding to haplotype I. The neutrality tests revealed that the COX2 gene was conserved throughout evolution, and the maximum likelihood phylogenetic analysis, using homologous sequences from other Ursidae species, showed clustering of the COX2 sequences of giant pandas, suggesting that this gene evolved differently in them. © 2017 The Authors.


Zhou X.,Sichuan Agricultural University | Yu H.,Sichuan Entry Exit Inspection and Quarantine Bureau | Wang N.,Sichuan Agricultural University | Xie Y.,Sichuan Agricultural University | And 9 more authors.
Journal of Wildlife Diseases | Year: 2013

The helminth Baylisascaris schroe-deri is one of the most harmful parasites infecting giant pandas (Ailuropoda melano-leuca). It is therefore important to develop an exact diagnostic technique to detect this parasite. Using a known number (1, 2, 3, 4, 5, 10, 25, 50, 100) of feces-isolated B. schroederi egg and adult DNA, we developed a PCR to detect a portion of the mitochondrial 12S rRNA and applied it to giant panda fecal samples. The method was sufficiently sensitive to detect B. schroederi DNA from isolated eggs in a fecal sample with a detection threshold of one egg. We detected B. schroederi in 88% of fecal samples, 30% higher than the conventional flotation technique. No cross-reactivity with other common nematode DNA was detected. Our PCR assay may constitute a valuable alternative for the diagnosis of B. schroederi infections. © Wildlife Disease Association 2013.


Wang X.-H.,Chengdu University of Technology | Wang X.-H.,Sichuan Agricultural University | Wang X.-H.,China Conservation and Research Center for Giant Panda | Huang Y.,Sichuan Agricultural University | And 5 more authors.
Chinese Journal of Applied Ecology | Year: 2010

In order to search for the ways to restrain the flowering process and promote the regeneration of bamboo (Phyllostachys heteroclada), five types of regeneration experiments were conducted, including 1) harvesting all the bamboos in the first year and the flowered bamboos in the next year all at the beginning stage of florescence, 2) harvesting the flowered bamboos at the beginning stage of florescence in two consecutive years, 3) harvesting the fully flowered bamboos in the first year and the bamboos at the beginning stage of florescence in the next year, 4) fertilizing the bamboo floor before growing season in consecutive two years, and 5) no any manipulation. In each of the cases, the carbon- and nitrogen metabolism and related enzyme activities in bamboo rhizome were studied. The results showed that treatment 1 had the best effect, which resulted in the greatest number (207) of new shoots produced and with the highest survival rate (69. 33% ), and had the most obvious effect on restraining bamboo flowering and promoting regeneration. Also, it produced the highest contents of soluble sugar (10. 89%) and total sugar (20. 39%), the highest total sugar to total nitrogen ratio (34. 56), and the highest glutamine synthetase activity (104. 52 nig · g -1 · h -1) in bamboo rhizome. These findings demonstrated that the carbon- and nitrogen metabolism in the rhizome had definite relationships with the flowering process and regeneration of bamboo, and the rhizome growth played a critical role in bamboo propagation.


Hu Y.-D.,Sichuan Agricultural University | Pang H.-Z.,Sichuan Agricultural University | Li D.-S.,China Conservation and Research Center for Giant Panda | Ling S.-S.,China Conservation and Research Center for Giant Panda | And 7 more authors.
Gene | Year: 2016

As the rate-limiting enzyme of the mitochondrial respiratory chain, cytochrome c oxidase (COX) plays a crucial role in biological metabolism. “Living fossil” giant panda (Ailuropoda melanoleuca) is well-known for its special bamboo diet. In an effort to explore functional variation of COX1 in the energy metabolism behind giant panda's low-energy bamboo diet, we looked at genetic variation of COX1 gene in giant panda, and tested for its selection effect. In 1545 base pairs of the gene from 15 samples, 9 positions were variable and 1 mutation leaded to an amino acid sequence change. COX1 gene produces six haplotypes, nucleotide (pi), haplotype diversity (Hd). In addition, the average number of nucleotide differences (k) is 0.001629 ± 0.001036, 0.8083 ± 0.0694 and 2.517, respectively. Also, dN/dS ratio is significantly below 1. These results indicated that giant panda had a low population genetic diversity, and an obvious purifying selection of the COX1 gene which reduces synthesis of ATP determines giant panda's low-energy bamboo diet. Phylogenetic trees based on the COX1 gene were constructed to demonstrate that giant panda is the sister group of other Ursidae. © 2016 Elsevier B.V.


Xie Y.,Sichuan Agricultural University | Zhou X.,Center for Animal Diseases Control and Prevention | Chen L.,Sichuan Agricultural University | Zhang Z.,Chengdu Research Base of Giant Panda Breeding | And 5 more authors.
Parasites and Vectors | Year: 2015

Background: Baylisascaris schroederi, an intestinal nematode of the giant panda, is the cause of the often fatal disease, baylisascariasis. Glutathione S-transferases (GSTs) are versatile enzymes that can affect parasite survival and parasite-host interactions and, are therefore, potential targets for the development of diagnostic tests and vaccines. Methods: In this study, we identified a full-length cDNA that encoded a novel, secretory sigma-like GST (Bsc-GSTσ) from a B. schroederi-omic dataset. Following cloning and sequencing, sequence and structural analyses and comparative modeling were performed using online-bioinformatics and proteomics tools. The recombinant Bsc-GSTσ (rBsc-GSTσ) protein was prokaryotically expressed and then used to detect antigenicity and reactivity using immunoblotting assays. In addition, the native protein in female adult B. schroederi was located via immunofluorescence techniques, while the preliminary ELISA-based serodiagnostic potential of rBsc-GSTσ was assessed in native and infected mouse sera. Results: Bsc-GSTσ contained a 621-bp open reading frame that encoded a polypeptide of 206 amino acids with two typical sigma GST domain profiles, including a GST-N-Sigma-like at the N-terminus and a GST-C-Sigma-like at the C-terminus. The presence of an N-terminal signal sequence indicated that Bsc-GSTσ was a secretory protein. Sequence alignment and phylogenetic analyses showed that Bsc-GSTσ was a nematode-specific member of the Sigma class GSTs and shared the closest genetic distance with its homologue in Ascaris suum. Further comparative structure analyses indicated that Bsc-GSTσ possessed the essential structural motifs (e.g., βαβαββα) and the consensus secondary or tertiary structure that is typical for other characterized GSTσs. Immunolocalization revealed strong distributions of native Bsc-GSTσ in the body hypodermis, lateral chords, gut epithelium, gut microvilli, oviduct epithelium, and ovaries of adult female worms, similar to its homologue in A. suum. Building on good immunogenic properties, rBsc-GSTσ-based ELISA exhibited a sensitivity of 79.1% and a specificity of 82.0% to detect anti-B. schroederi IgG antibodies in the sera of experimentally infected mice. Conclusion: This study presents a comprehensive demonstration of sequence and structural-based analysis of a new, secretory sigma-like GST from a nematode, and its good serodiagnostic performance suggests that rBsc-GSTσ has the potential to detect B. schroederi and, therefore, could be used to develop an ELISA-based serological test to diagnose baylisascariasis in giant pandas. © 2015 Xie et al.; licensee BioMed Central.


Gu Y.,Sichuan Agricultural University | Wang Y.,Sichuan Agricultural University | Ma X.,Sichuan Agricultural University | Ma X.,Nanjing Agricultural University | And 14 more authors.
Applied Biochemistry and Biotechnology | Year: 2015

While taxol yields of fungi from non-animal sources are still low, whether Pestalotiopsis hainanensis isolated from the scurf of a dermatitic giant panda, Ailuropoda melanoleuca, provides a greater taxol yield remains unknown. The objective of the study was to determine the corresponding taxol yield. The structure of the taxol produced by the fungus was evaluated by thin layer chromatography (TLC), ultraviolet (UV) spectroscopy, high-performance liquid chromatography (HPLC), 1H and 13C nuclear magnetic resonance spectroscopy (1H-NMR and 13C-NMR), and time-of-flight mass spectrometry (TOF-MS), with standard taxol as a control. The results demonstrated that the P. hainanensis fungus produced taxol, which had the same structure as the standard taxol and yield of 1,466.87 μg/L. This fungal taxol yield from the dermatitic giant panda was significantly greater than those of fungus from non-animal sources. The taxol-producing fungus may be a potential candidate for the production of taxol on an industrial scale. © 2014, Springer Science+Business Media New York.


PubMed | China Conservation and Research Center for Giant Panda and Sichuan Agricultural University
Type: Journal Article | Journal: Gene | Year: 2016

As the rate-limiting enzyme of the mitochondrial respiratory chain, cytochrome c oxidase (COX) plays a crucial role in biological metabolism. Living fossil giant panda (Ailuropoda melanoleuca) is well-known for its special bamboo diet. In an effort to explore functional variation of COX1 in the energy metabolism behind giant pandas low-energy bamboo diet, we looked at genetic variation of COX1 gene in giant panda, and tested for its selection effect. In 1545 base pairs of the gene from 15 samples, 9 positions were variable and 1 mutation leaded to an amino acid sequence change. COX1 gene produces six haplotypes, nucleotide (pi), haplotype diversity (Hd). In addition, the average number of nucleotide differences (k) is 0.0016290.001036, 0.80830.0694 and 2.517, respectively. Also, dN/dS ratio is significantly below 1. These results indicated that giant panda had a low population genetic diversity, and an obvious purifying selection of the COX1 gene which reduces synthesis of ATP determines giant pandas low-energy bamboo diet. Phylogenetic trees based on the COX1 gene were constructed to demonstrate that giant panda is the sister group of other Ursidae.


Prescott H.M.A.,Durham University | Manning C.,Durham University | Gardner A.,Durham University | Ritchie W.A.,Roslin Embryology Ltd. | And 5 more authors.
PLoS ONE | Year: 2015

Since the first mammal was cloned, the idea of using this technique to help endangered species has aroused considerable interest. However, several issues limit this possibility, including the relatively low success rate at every stage of the cloning process, and the dearth of usable tissues fromthese rare animals. iPS cells have been produced from cells from a number of rare mammalian species and this is the method of choice for strategies to improve cloning efficiency and create new gametes by directed differentiation. Nevertheless information about other stem cell/progenitor capabilities of cells from endangered species could prove important for future conservation approaches and adds to the knowledge base about cellular material that can be extremely limited. Multipotent progenitor cells, termed skin-derived precursor (SKP) cells, can be isolated directly from mammalian skin dermis, and human cheek tissue has also been shown to be a good source of SKP-like cells. Recently we showed that structures identical to SKPs termedm-SKPs could be obtained from monolayer/two dimensional (2D) skin fibroblast cultures. Here we aimed to isolate m-SKPs fromcultured cells of three endangered species; giant panda (Ailuropoda melanoleuca); red panda (Ailurus fulgens); and Asiatic lion (Panthera leo persica). m-SKP-like spheres were formed from the giant panda buccalmucosa fibroblasts; whereas dermal fibroblast (DF) cells cultured fromabdominal skin of the other two species were unable to generate spheres. Under specific differentiation culture conditions giant panda spheres expressed neural, Schwann, adipogenic and osteogenic cell markers. Furthermore, these buccal mucosa derived spheres were shown to maintain expression of SKP markers: nestin, versican, fibronectin, and P75 and switch on expression of the stemcellmarker ABCG2. These results demonstrate that giant panda cheek skin can be a useful source ofm-SKP multipotent progenitors. At present lack of sample numbers means that we can only postulate why we were unable to obtainm-SKPs from the lion and red panda cultures. However the giant panda observations point to the value of archiving cells from rare species, and the possibilities for later progenitor cell derivation. © 2015 Prescott et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


PubMed | Royal Zoological Society of Scotland, Durham University, Roslin Embryology Ltd. and China Conservation and Research Center for Giant Panda
Type: Journal Article | Journal: PloS one | Year: 2015

Since the first mammal was cloned, the idea of using this technique to help endangered species has aroused considerable interest. However, several issues limit this possibility, including the relatively low success rate at every stage of the cloning process, and the dearth of usable tissues from these rare animals. iPS cells have been produced from cells from a number of rare mammalian species and this is the method of choice for strategies to improve cloning efficiency and create new gametes by directed differentiation. Nevertheless information about other stem cell/progenitor capabilities of cells from endangered species could prove important for future conservation approaches and adds to the knowledge base about cellular material that can be extremely limited. Multipotent progenitor cells, termed skin-derived precursor (SKP) cells, can be isolated directly from mammalian skin dermis, and human cheek tissue has also been shown to be a good source of SKP-like cells. Recently we showed that structures identical to SKPs termed m-SKPs could be obtained from monolayer/ two dimensional (2D) skin fibroblast cultures. Here we aimed to isolate m-SKPs from cultured cells of three endangered species; giant panda (Ailuropoda melanoleuca); red panda (Ailurus fulgens); and Asiatic lion (Panthera leo persica). m-SKP-like spheres were formed from the giant panda buccal mucosa fibroblasts; whereas dermal fibroblast (DF) cells cultured from abdominal skin of the other two species were unable to generate spheres. Under specific differentiation culture conditions giant panda spheres expressed neural, Schwann, adipogenic and osteogenic cell markers. Furthermore, these buccal mucosa derived spheres were shown to maintain expression of SKP markers: nestin, versican, fibronectin, and P75 and switch on expression of the stem cell marker ABCG2. These results demonstrate that giant panda cheek skin can be a useful source of m-SKP multipotent progenitors. At present lack of sample numbers means that we can only postulate why we were unable to obtain m-SKPs from the lion and red panda cultures. However the giant panda observations point to the value of archiving cells from rare species, and the possibilities for later progenitor cell derivation.


Kersey D.C.,Smithsonian Conservation Biology Institute | Kersey D.C.,Western University of Health Sciences | Wildt D.E.,Smithsonian Conservation Biology Institute | Brown J.L.,Smithsonian Conservation Biology Institute | And 3 more authors.
Reproduction, Fertility and Development | Year: 2010

The aim of the present study was to determine the efficacy of faecal hormonal measures for evaluating ovarian activity in a significant sized cohort of giant pandas during the perioestrual period. Faecal excretion of oestrogen and progestagen metabolites corresponded with urinary patterns and receptive behaviours. Longitudinal assessment of 10 females revealed that, on average, faecal oestrogen concentrations started to rise (P<0.05) above baseline (baseline means.e.m.; 64.7 ± 6.6ngg-1) 5 days before the preovulatory oestrogen peak (484.6±126.8ngg-1), which was followed by a gradual descent over 4 days to nadir. Mean faecal progestagen metabolite concentrations increased approximately twofold above baseline (from 186.2 ± 37.7 to 347.2 ± 75.7ngg-1; P<0.05) during the 20-day interval after the preovulatory oestrogen surge. Variability within and among females precluded the use of a threshold of oestrogen or progestagen metabolites to predict reproductive status, yet faeces collected 23 days per week provided sufficient data to recognise that an individual was in the perioestrual period. Finally, in females that were examined for at least 3 consecutive years, there was an 18-53 day variation in the onset and an 813 day variation in the duration of perioestrual behaviour from year to year. In summary, these findings indicate that gonadal hormone profiles associated with the period immediately before, during and after oestrus are accurately revealed by analysis of the fibrous faeces of the giant panda. This approach has potential value for providing point-in-time information on the reproductive status of free-living individuals. © 2010 CSIRO.

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