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Wang W.,Sichuan Animal Science Academy | Yan H.,Chengdu Zoo | Yu J.,Chengdu Zoo | Yi J.,Sichuan Animal Science Academy | And 5 more authors.
PLoS ONE | Year: 2017

The hog deer (Axis porcinus) is a small deer whose natural habitat is the wet or moist tall grasslands in South and Southeast Asia. Wild populations have dramatically decreased in recent decades. While wild hog deer were recently acknowledged to be extinct in China, a few captive populations have been maintained. In the present study, we successfully employed the restriction-site-associated DNA sequencing (RAD-seq) technique to generate a genome-wide profile of single-nucleotide polymorphisms (SNPs) in the captive population of hog deer from Chengdu Zoo, China (N = 11). Up to 4.7 million clean reads per sample were sequenced, and 11,155 SNPs and 8,247 haplotypes were simultaneously observed within more than 80% of sequenced individuals. In this population, the mean frequency of major alleles at each polymorphism site was 0.7903±0.0014, and the average nucleotide diversity (p) and inbreeding coefficient (FIS) were 0.3031±0.0015 and-0.0302±0.0062, respectively. Additionally, the Euclidean distance-based multidimensional scaling method revealed that the pairwise genetic relatedness was evenly distributed. However, the results of homologous searching by short reads did not provide any meaningful explanation of the phylogenetic relationship of hog deer, which should be further investigated. In conclusion, our results revealed current state of genetic diversity in this captive population of hog deer. Furthermore, these genome-wide SNPs would be useful for guiding the mating schedule to avoid sharp increase of inbreeding coefficient. © 2017 Wang 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.


Lin X.,CAS Chengdu Institute of Biology | Huang Y.,Guangxi Botanical Garden oj Medicinal Plants | Zuo Z.,Chengdu Zoo | Chen Y.,CAS Chengdu Institute of Biology
Chinese Journal of Applied and Environmental Biology | Year: 2012

Among many hypotheses proposed to explain large-scale patterns of species richness, three alternative versions of ecological hypothesis, the species-energy, habitat heterogeneity and contemporary climate hypotheses have shown widespread empirical support as the primary determinants of species richness. The database of 26 environmental variables, indicators of the different hypotheses, and human density and crop-agricultural area, indicators of the intensity of human impacts were used to explore large-scale patterns of species richness and their environmental determinants for primate in Mainland China using eigenvector-based spatial filters models (SEVM) at 100 km×100 km, 200 km×200 km and 400 km×400 km spatial resolution. The results suggested that primate species richness varied significantly on latitudinal gradients, decreasing from south to north. Species richness peaked mainly in mountain province, including western Yunnan and borderland province in southern Yunnan in southern China, but few species richness were found in the northeastern China, northern China, Mongolia-Xinjiang China and Qinghai-Tibet China at all spatial resolutions. At the different resolutions, the primary predictors of large-scale primate species richness patterns in Mainland China were not all the same. However, ambient-energy hypothesis (indicated by potential evapotranspiration) and climate hypothesis (indicated by annual mean temperature) may work together and best explain patterns of primate species richness in Mainland China at different spatial resolution. Our findings showed that the intensity of human impacts had effects on large-scale patterns of primate species richness when considering the intensity of human impacts together with environmental variables. Our results provide the necessary conservation implications for habitat loss of primate species richness.


Zhang W.,Chengdu Research Base of Giant Panda Breeding | Yue B.,University of Sichuan | Wang X.,University of Sichuan | Zhang X.,University of Sichuan | And 16 more authors.
Molecular Biology Reports | Year: 2011

In order to investigate the mitochondrial genome of Panthera tigris amoyensis, two South China tigers (P25 and P27) were analyzed following 15 cymt-specific primer sets. The entire mtDNA sequence was found to be 16,957 bp and 17,001 bp long for P25 and P27 respectively, and this difference in length between P25 and P27 occurred in the number of tandem repeats in the RS-3 segment of the control region. The structural characteristics of complete P. t. amoyensis mitochondrial genomes were also highly similar to those of P. uncia. Additionally, the rate of point mutation was only 0.3% and a total of 59 variable sites between P25 and P27 were found. Out of the 59 variable sites, 6 were located in 6 different tRNA genes, 6 in the 2 rRNA genes, 7 in non-coding regions (one located between tRNA-Asn and tRNA-Tyr and six in the D-loop), and 40 in 10 protein-coding genes. COI held the largest amount of variable sites (9 sites) and Cytb contained the highest variable rate (0.7%) in the complete sequences. Moreover, out of the 40 variable sites located in 10 protein-coding genes, 12 sites were nonsynonymous. © 2010 Springer Science+Business Media B.V.


Yan Y.,Sichuan University | Niu L.,Chengdu Zoo | Deng J.,Chengdu Zoo | Wang Q.,Chengdu Zoo | And 6 more authors.
Journal of Veterinary Medical Science | Year: 2015

Canine distemper virus (CDV) is a morbillivirus known to cause morbidity and mortality in a broad range of animals. Giant pandas (Ailuropoda melanoleuca), especially captive ones, are susceptible to natural infection with CDV. Interleukin-18 (IL-18) is a powerful adjuvant molecule that can enhance the development of antigen-specific immunity and vaccine efficacy. In this study, a giant panda IL-18 gene eukaryotic expression plasmid (pcAmIL-18) was constructed. Female BALB/c mice were muscularly inoculated with the plasmids pcAmIL-18, pcDNA3.1 and PBS, respectively. They were subsequently injected with an attenuated CDV vaccine for dogs, and the induced humoral and cellular responses were evaluated. The results showed that pcAmIL-18 remarkably improved the level of specific antibody, IFN-γ and IL-2 in mice sera, the T lymphocyte proliferation index and the percentage of CD4+ and CD8+ cells. These data indicated that pcAmIL-18 is a potential adjuvant that promotes specific immunity. ©2015 The Japanese Society of Veterinary Science.


Yu J.,Chengdu Zoo | Wang Q.,Chengdu Zoo | Liu X.,Chengdu Zoo | Deng J.,Chengdu Zoo | And 4 more authors.
Acta Theriologica Sinica | Year: 2010

The hog deer (Axis porcinus) is a Class 1 National Key Protected Species in China ; the species has been extirpated within China. China has a captive population of only about 30, of which the Chengdu Zoo has 22. It is critical for captive management within China to identify paternity relationships and establish the pedigree of the hog deer population in the Chengdu Zoo. We used 7 microsatellites to genotype 22 live and 5 dead hog deer individuals from the Chengdu Zoo ; we identified potential 13 paternal relationships for animals in which maternity was already known. The exclusion method defined 8 of the 13 father - offspring pairs. The likelihood method determined the remaining 5 pairs with a confidence level of 95%. Combining the result of this paternity test with the breeding records of the Chengdu Zoo, we constructed the genetic pedigree for this hog deer population. Our result will serve as an important reference for the future artificial breeding strategy and management of the captive hog deer population in China.


Yan Y.,Sichuan University | Wang Q.,Chengdu Zoo | Niu L.L.,Chengdu Zoo | Deng J.B.,Chengdu Zoo | And 4 more authors.
Genetics and molecular research : GMR | Year: 2014

Interleukin 18 (IL-18), as a member of IL-1 superfamily, is an important pleiotropic cytokine that modulates Th1 immune responses. In this report, we cloned and identified a homolog of IL-18 in giant panda (Ailuropoda melanoleuca) (designated as AmIL-18) from peripheral blood mononuclear cells stimulated with lipopolysaccharide. The open readin g frame of AmIL-18 cDNA is 579 bp encoding a deduced protein of 192 amino acids. AmIL-18 gDNA fragments contained 5 exons and 4 introns. The amino acid sequence of AmIL-18 shared 23.9 to 87.0% identity with other species. To evaluate the effects of AmIL-18 on the immune response, we expressed the recombinant AmIL-18 in Escherichia coli BL21 (DE3). The fusion protein PET-AmIL-18 was purified by nickel affinity column chromatography and verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. The biological function of purified PET-AmIL-18 was determined on mouse splenocytes by quantitative real-time polymerase chain reaction. INF-γ and other cytokines were increased when stimulated by PET-AmIL-18, particularly when combined with recombinant human interleukin 12, while a Th2-type cytokine, interleukin-4, was strikingly suppressed. These results will provide information for the potential use of recombinant proteins to manipulate the immune response in giant pandas and facilitate the study to protect this treasured species.


Yan Y.,Sichuan University | Wang Q.,Chengdu Zoo | Niu L.L.,Chengdu Zoo | Deng J.B.,Chengdu Zoo | And 5 more authors.
Genetics and Molecular Research | Year: 2014

Interleukin 18 (IL-18), as a member of IL-1 superfamily, is an important pleiotropic cytokine that modulates Th1 immune responses. In this report, we cloned and identified a homolog of IL-18 in giant panda (Ailuropoda melanoleuca) (designated as AmIL-18) from peripheral blood mononuclear cells stimulated with lipopolysaccharide. The open reading frame of AmIL-18 cDNA is 579 bp encoding a deduced protein of 192 amino acids. AmIL-18 gDNA fragments contained 5 exons and 4 introns. The amino acid sequence of AmIL-18 shared 23.9 to 87.0% identity with other species. To evaluate the effects of AmIL-18 on the immune response, we expressed the recombinant AmIL-18 in Escherichia coli BL21 (DE3). The fusion protein PET-AmIL-18 was purified by nickel affinity column chromatography and verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. The biological function of purified PET-AmIL-18 was determined on mouse splenocytes by quantitative real-time polymerase chain reaction. INF-γ and other cytokines were increased when stimulated by PET-AmIL-18, particularly when combined with recombinant human interleukin 12, while a Th2-type cytokine, interleukin-4, was strikingly suppressed. These results will provide information for the potential use of recombinant proteins to manipulate the immune response in giant pandas and facilitate the study to protect this treasured species. © FUNPEC-RP.


PubMed | Chengdu Zoo and Sichuan University
Type: | Journal: Research in veterinary science | Year: 2016

The giant panda (Ailuropoda melanoleuca) is an endangered species. Interleukin-18 (IL-18) plays an important role in the innate and adaptive immune responses by inducing IFN-. IL-18 has been implicated in the pathogenesis of various diseases. IL-18 binding protein (IL-18BP) is an intrinsic inhibitor of IL-18 that possesses higher affinity to IL-18. In this study, we cloned and characterized IL-18BP in giant panda (AmIL-18BP) from the spleen. The amino acid sequence of giant panda IL-18BP ORF shared about 65% identities with other species. To evaluate the effects of AmIL-18BP on the immune responses, we expressed the recombinant AmIL-18BP in Escherichia coli BL21 (DE3).The fusing protein PET-AmIL-18BP was purified by nickel affinity column chromatography. The biological function of purified PET-AmIL-18BP was determined on mice splenocyte by qRT-PCR. The results showed that AmIL-18BP was functional and could significantly reduce IFN- production in murine splenocytes. These results will facilitate the study of protecting giant panda on etiology and immunology.


PubMed | Chengdu Zoo and Sichuan University
Type: Journal Article | Journal: Genetics and molecular research : GMR | Year: 2014

Interleukin 18 (IL-18), as a member of IL-1 superfamily, is an important pleiotropic cytokine that modulates Th1 immune responses. In this report, we cloned and identified a homolog of IL-18 in giant panda (Ailuropoda melanoleuca) (designated as AmIL-18) from peripheral blood mononuclear cells stimulated with lipopolysaccharide. The open readin g frame of AmIL-18 cDNA is 579 bp encoding a deduced protein of 192 amino acids. AmIL-18 gDNA fragments contained 5 exons and 4 introns. The amino acid sequence of AmIL-18 shared 23.9 to 87.0% identity with other species. To evaluate the effects of AmIL-18 on the immune response, we expressed the recombinant AmIL-18 in Escherichia coli BL21 (DE3). The fusion protein PET-AmIL-18 was purified by nickel affinity column chromatography and verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. The biological function of purified PET-AmIL-18 was determined on mouse splenocytes by quantitative real-time polymerase chain reaction. INF- and other cytokines were increased when stimulated by PET-AmIL-18, particularly when combined with recombinant human interleukin 12, while a Th2-type cytokine, interleukin-4, was strikingly suppressed. These results will provide information for the potential use of recombinant proteins to manipulate the immune response in giant pandas and facilitate the study to protect this treasured species.

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