吴孝兵
扬子鳄保护生物学
个性化签名
- 姓名:吴孝兵
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学术头衔:
博士生导师
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学科领域:
动物学
- 研究兴趣:扬子鳄保护生物学
吴孝兵,博士,男,1965年2月生,安徽全椒人。现为安徽师范大学教授,博士生导师,“皖江学者”计划讲席教授,安徽省首批跨世纪学科带头人培养对象。现兼任中国动物学会两栖爬行动物学专业委员会副主任,安徽省动物学会副理事长,国际自然保护联盟鳄类专家组(IUCN/CSG)成员,中国生态学会动物生态学专业委员会委员,中华人民共和国濒危物种科学委员会协审专家,《动物学杂志》编委。十多年来主要致力于扬子鳄保护生物学的研究工作外,还积累了一些野外调查工作经验,近年来主持过“安徽省宁国万家拟建自然保护区资源调查”、“泾县汀溪自然保护区资源调查”、“扬子鳄野生种群数量调查”及“安徽扬子鳄国家级自然保护区综合科学考察”,参加四川龙溪-虹口国家级自然保护区的科考及“安徽清凉峰自然保护区”的科考工作等。曾在《Biological Conservation》,《Chinese Science Bulletin》等刊物发表第一作者或通讯作者学术论文80多篇,合作出版专著《扬子鳄研究》1部。目前主持国家自然科学基金2项、安徽省优秀青年基金项1项、安徽省自然科学基金等省部级研究项目多项;获安徽省科学技术奖(自然科学类)二等奖(主持)等,并于2005年获安徽省政府特殊津帖。现指导在读博士生4名,硕士生19名。
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吴孝兵, Xiao-Bing Wu a, b, *, Yi-Quan Wang a, Kai-Ya Zhou a, Wei-Quan Zhu a, Ji-Shan Nie c, Chao-Lin Wang c, Wan-Shu Xie c
Biological Conservation 106(2002)435-441,-0001,():
-1年11月30日
Chinese alligator (Alligator sinensis) is an endemic species in China. The likely extinction of it in the wild has been recognised. To prevent this species becoming extinct, the Anhui Research Centre of Chinese Alligator Reproduction (ARCCAR) was established in Xuanzhou, Anhui Province in 1979, where has been established the largest captive population of Chinese alligator (XZSP) in the world. Another farm (CXSP) was established by villagers in Changxin, Zhejiang Province. The results of an investigation of the two captive subpopulation structures by genetic analysis are presented in this paper. We examined the genetic variation in the two captive subpopulations using RAPDs. Thirty-one random primers were selected among 199 random primers screened. A total of 193 reproducible RAPD fragments were scored among 43 individuals, of which 21 (10.88%) were polymorphic. The genetic distances between 43 individuals ranged from 0 to 0.0376 with average of 0.0104 0.0055 S. E. The genetic similarity in CXSP (0.9948 0.0029 S. E.) was higher than that in XZSP (0.9894 0.0055 S.E.). The founder effect is a possible explanation for very low genetic variation in CXSP. Analysis of the RAPD data showed that the mean phenotypic band frequencies of each polymorphic loci was 0.6656 0.3730 S. E. The lowest phenotypic band frequency (0.0233) was found in four of those polymorphic loci. There was no genetic difference between the two subpopulations (Dij=0.0009). According to the dendrogram and the distribution of polymorphic fragments in two subpopulations, CXSP originated genetically from XZSP. This paper summarises a preliminary research on genetic structure in populations of Chinese alligator. Although there is higher genetic similary (0.9896 0.0055 S. E.) in captive population of A. sinensis, we did not determine whether or not loss of genetic variation had occurred in relation to a wild control population. The data of malformed offspring will be collected carefully, and wild samples be added to set up a control population in future study.
Alligator sinensis, RAPD, Genetic variation, Ex sito conservation
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吴孝兵, WU Xiaobing, , WANG Yiquan, ZHOU Kaiya, ZHU Weiquan, NIE Jishan & WANG Chaolin
Chinese Science Bulletin 48(2003)2050-2054,-0001,():
-1年11月30日
The 16746-neucleotide (nt) sequence of mitochondrial DNA (mtDNA) of Chinese alligator, Alligator sinensis, was determined using the Long-PCR and primer walking methods. As is typical in vertebrates, the mtDNA encodes 13 proteins, 2 rRNA, 22 tRNA genes, and a noncoding control region. The composition of bases is respectively 29.43% A, 24.59% T, 14.86% G, 31.12% C. The gene arrangement differs from the common vertebrate gene arrangement, but is similar to that of other crocodiles. DNA sequence data from 12S rRNA, 16S rRNA, protein-coding genes and combined sequence data were used to reconstruct the phylogeny of reptiles with the MP and MLmethods. With this large data set and an appropriate range of outgroup taxa, the authors demonstrate that Chinese alligator is most closely related to American alligator among three crocodilian species, which suppors the traditional viewpoint. According to the branch lengths of ML tree from the combined data set, the primary divergence between Alligator and Caiman genus was dated at about 74.9 Ma, the split between Chinese alligator and American alligator was dated at 50.9 Ma.
Chinese alligator, mitochondrial genome, complete sequence, phylogeny, divergence time
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吴孝兵, Peng Yan a, Xiao-Bing Wu a, *, Yan Shi a, Chang-Ming Gu b, Ren-Ping Wang c, Chao-Ling Wang c
Biological Conservation 121(2005)45-51,-0001,():
-1年11月30日
With the commercial farming and exploitation of Chinese alligators (Alligator sinensis), illegal and inappropriately labeled Chinese alligator meat has appeared in markets. To prevent the illegal hunting and commerce for Chinese alligators, it will be important to develop an expedient and practical method for the identification of Chinese alligator meat. In this study, a pair of the species-specific PCR primers (Alli-M and Alli-R) was designed using sequence variations of mitochondrial DNA (mtDNA) cytochrome b gene between Chinese alligators and other crocodilians. By the multiplex PCR of using the species-specific primers and 12S rRNA universal primers L1091 and H1478, 31 samples (27 meat samples, 4 skin samples) were identified. The result of amplification displayed that only the fresh and the cooked meat samples from the Chinese alligator could be amplified with two bands. We also present a case of identification of a crocodilian body part found in a local market using the newly developed primers. The specific primers designed in this study could be widely used for the rapid and accurate identification of not only alligator meat but also other commercial products from Chinese alligator.
Chinese alligator, Meat, Identification, Species-specific primers, Multiplex PCR
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【期刊论文】Immunohistochemical study on gastrointestinal endocrine cells of four reptiles
吴孝兵, Xu-Gen Huang, Xiao-Bing Wu
World J Gastroenterol 11(2005)5498-5505,-0001,():
-1年11月30日
AIM: To clarify the types, regional distributions and distribution densities as well as morphological features of gastrointestinal (GI) endocrine cells in various parts of the gastrointestinal track (GIT) of four reptiles, Gekko japonicus, Eumeces chinensis, Sphenomorphus indicus and Eumeces elegans. METHODS: Paraffin-embedded sections (5μm) of seven parts (cardia, fundus, pylorus, duodenum, jejunum, ileum, rectum) of GIT dissected from the four reptiles were prepared. GI endocrine cells were revealed by using immunohistochemical techniques of streptavidin-peroxidase (S-P) method. Seven types of antisera against 5-hydroxytryptamine (5-HT), somatostatin (SS), gastrin (GAS), glucagon (GLU), substance P (SP), insulin and pancreatic polypeptide were identified and then GI endocrine cells were photomicrographed and counted. RESULTS: The GI endocrine system of four reptiles was a complex structure containing many endocrine cell types similar in morphology to those found in higher vertebrates. Five types of GI endocrine cells, namely 5-HT, SS, GAS, SP and GLU immunoreactive (IR) cells were identified in the GIT of G. japonicus, E. chinensis and S. indicus; while in the GIT of E. elegans only the former three types of endocrine cells were observed. No PP- and INS- IR cells were found in all four reptiles. 5-HT-IR cells, which were most commonly found in the pylorus or duodenum, distributed throughout the whole GIT of four reptiles. However, their distribution patterns varied from each other. SS-IR cells, which were mainly found in the stomach especially in the pylorus and/or fundus, were demonstrated in the whole GIT of E. chinensis, only showed restricted distribution in the other three species. GAS-IR cells, with a much restricted distribution, were mainly demonstrated in the pylorus and/or the proximal small intestine of four reptiles. GLU-IR cells exhibited a limited and species-dependent variant distribution in the GIT of four reptiles. SP-IR cells were found throughout the GIT except for jejunum in E. elegans and showed a restricted distribution in the GIT of G. japonicus and S. indicus. In the GIT of four reptiles the region with the highest degree of cell type heterogeneity was pylorus and most types of GI endocrine cells along the GIT showed the peak density in pylorus as well. CONCLUSION: Some common and unique features of the distribution and morphology of different types of GI endocrine cells are found in four reptiles. This common trait may reflect the similarity in digestive physiology of various vertebrates.
Immunohistochemistry, Gastrointestinal tract, Endocrine cells, Distribution patterns, Four reptiles
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吴孝兵, XIAOBING WU, HUI LIU
New Zealand Journal of Zoology 33(2006)65-71,-0001,():
-1年11月30日
To distinguish sika deer (Cervus nippon) tissue samples from those of sympatric cervids in the southern part of Anhui Province, China, we analysed randomly amplified polymorphic DNA (RAPD) fragments of 12 individual samples from four cervid species: black muntjac (Muntiacus crinifrons), Reeve’s muntjac (Muntiacus reevesi), sika deer (Cervus nippon), and tufted deer (Elaphodus cephalophus). Only one primer among the 100 screened produced a clear specific band for identifying DNA of sika deer. We cloned and sequenced 449 bp from this fragment of DNA. We then designed a pair of 18 bp primers (MHL-U/MHL-D) according to the sequence, resulting in a 251 bp sequence-characterised amplified region (SCAR) for sika deer. By combining this pair of SCAR primers with a universal set of mammalian DNA primers, the identification of sika deer tissue samples by a simple common multiplex PCR assay is straightforward, rapid, and reliable. The method will be useful for cervid conservation and cervid bushmeat trade regulation.
Cervus nippon, tissue identification, SCAR, multiplex PCR
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吴孝兵, X. B. Wu, H. Xue, L. S. Wu, J. L. Zhu & R. P. Wang
Animal Biodiversity and Conservation Vol. 29 No.1 (2006) 65-71,-0001,():
-1年11月30日
Regression analysis between body and head measurements of Chinese alligators (Alligator sinensis) in the captive population.— Four body–size and fourteen head–size measurements were taken from each Chinese alligator (Alligator sinensis) according to the measurements adapted from Verdade. Regression equations between body–size and head–size variables were presented to predict body size from head dimension. The coefficients of determination of captive animals concerning body–and head–size variables can be considered extremely high, which means most of the head–size variables studied can be useful for predicting body length. The result of multivariate allometric analysis indicated that the head elongates as in most other species of crocodilians. The allometric coefficients of snout length (SL) and lower ramus (LM) were greater than those of other variables of head, which was considered to be possibly correlated to fights and prey. On the contrary, allometric coefficients for the variables of obita (OW, OL) and postorbital cranial roof (LCR), were lower than those of other variables.
Regression analysis, Allometry, Chinese alligator
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吴孝兵, 吴孝兵①③, 王义权①②*, 周开亚①, 朱伟栓①, 聂继山④, 王朝林④
科学通报,2003,48(18):1954~1958,-0001,():
-1年11月30日
通过酶切、克隆、测序、结合Long-PCR和PrimerWalking法对扬子鳄线粒体DNA基因组进行全序列测定,结果表明,扬子鳄线粒体基因组DNA序列全长为16746bp,其基因组碱基组成为29.43%A,24.59%T,14.86%G,31.12%C.与其他大多数脊椎动物相同,其基因组由13个蛋白质编码基因、2个rRNA基因、22个tRNA基因及1个非编码的控制区(D-loop)组成,基因的排列与已测序的鳄类相似在全序列分析的基础上,对12SrRNA基因序列、16SrRNA基因序列、蛋白质编码基因序列及其合并数据用MP法和ML法构建系统发生树,结果表明扬子鳄与密河鳄的亲缘关系较近,支持传统观点根据树的支长数值估计钝吻鳄属发生的时间为74.9MaBP,扬子鳄与密河鳄分歧的时间为50.9MaBP.
扬子鳄, 线粒体基因组, 全序列, 系统发生, 分歧时间
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吴孝兵, 史燕, 吴孝兵**, 晏鹏, 赵哲
动物学报,2004,50(2):297~301,-0001,():
-1年11月30日
运用一种改进的提取方法,作者从鞣制皮革中成功地提取了总DNA,同时还对尾尖皮、鳞片、盐腌生皮等皮质进行了DNA提取;用12SrRNA基因扩增的通用引物、扬子鳄鉴别引物、微卫星引物及RAPD引物进行PCR扩增,并对部分扩增结果进行测序,以检验提取效果。结果证明,几种皮质标本都可提取出DNA,其中尾尖皮和鳞片的提取效果较好,用四种引物都可扩增出明显亮带;盐腌生皮和鞣制皮提取结果也很好,并且用12SrRNA通用引物、扬子鳄鉴别引物扩增的亮带较明显,可进行扬子鳄皮质用品等的分子鉴定及部分序列的扩增和测序研究[动物学报50(2):297-301,2004]。
扬子鳄, 皮革, 总DNA, PCR
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【期刊论文】扬子鳄MHC Ⅱ类B基因第二外元的克隆及序列分析
吴孝兵, 史燕, 吴孝兵*, 晏鹏, 陈壁辉
动物学研究,2004,25(5):415~421,-0001,():
-1年11月30日
3头扬子鳄血样取自宣城安徽省扬子鳄繁殖研究中心。利用一对简并引物对MHCⅡ类B基因第二外元的部分片段进行扩增;通过克隆、单链构象多态性分析、测序,并将测得序列与下载的8个物种MHC序列比对,确定序列差异和变异位点;利用MEGA软件构建NJ树,PAUP4.0构建MP树。结果得到10种不同的序列,片段长166bp。核苷酸序列中有38个变异位点,氨基酸序列中有23个变异位点;推定的抗原结合位点非同义替换(dN)明显高于同义替换(dS)。10种序列的NJ树和MP树极为相似,均为A、B两个分支,两个分支明显的特异性位点核苷酸序列中有9个,氨基酸序列中有7个。表明扬子鳄MHCⅡ类B基因第二外元有较高的多态性,有利于扬子鳄饲养种群的遗传保护。
扬子鳄, MHC Ⅱ类B基因第二外元, 序列分析
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吴孝兵, 吴陆生, 吴孝兵*, 江红星, , 王朝林
生物多样性,2005,13(2):156~161,-0001,():
-1年11月30日
作者分别于2002和2003年抽样调查了安徽省扬子鳄国家级自然保护区有或曾经有野生扬子鳄(Alligatorsinensis)分布的22个样地,选择了与野生扬子鳄生存有关的8类生态因子,即水域中岛屿情况、水域水面的稳定度、水体pH值、螺类丰富度、岸线植被盖度、岸线土壤质地、苦竹密度和植被类型,运用资源选择函数结合主成分分析方法研究了野生扬子鳄对生境的选择。结果表明岸线植被盖度对野生扬子鳄的生境选择影响最大,其次是水体pH值,再次是螺类丰富度、苦竹密度、水域水面的稳定度和土壤质地;而水域中岛屿情况与植被类型对野生扬子鳄生境选择的影响则较弱。
Alligator sinensis, 生境特征, 资源选择函数, 主成分分析
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