大熊猫肠道纤维素分解菌的分离鉴定及产酶性质

【目的】从健康大熊猫新鲜粪便中分离具有纤维素酶活性的菌株,并对其进行菌种鉴定及产酶性质研究。【方法】利用羧甲基纤维素钠培养基分离纯化具有较高纤维素酶活性的菌株,根据形态学特征、生理生化特性以及16S rDNA分析对其进行分类鉴定,研究影响该菌株纤维素酶的产酶条件,以及对不同纤维素底物的降解情况。【结果】分离得到一株纤维素酶产生菌株P2,该菌株为好氧的革兰氏阳性细菌,生长温度范围20-50℃(最适温度37℃),pH范围6.0-9.0(最适pH7.0),NaCl浓度范围0%-15%(最适2%NaCl),培养24h达到产酶高峰。16S rDNA基因序列分析显示,菌株P2与解淀粉芽胞杆菌(Bacillusamyloliquefaciens)NBRC15535相似性为99.66%。该菌株对四种纤维素底物(滤纸、脱脂棉、秸秆、竹纤维)均有不同程度的降解,内切葡聚糖酶、外切葡聚糖酶、β-葡萄糖苷酶和总酶活具有不同的酶活变化。【结论】本研究首次从大熊猫粪便中分离出了好氧纤维素分解菌,并鉴定为解淀粉芽胞杆菌,对上述四种纤维结构均有一定的破坏和分解作用,为进一步研究大熊猫竹纤维消化机制提供了菌源。     

汶川地震对四川龙溪—虹口和唐家河自然保护区大熊猫栖息地利用格局的影响

2008年的汶川地震发生在大熊猫集中分布的区域,对大熊猫的栖息地造成了严重破坏.利用龙溪—虹口和唐家河两个国家级自然保护区地震前(2003～ 2007)和地震后(2008～ 2010)共计7年的大熊猫监测数据,就汶川地震对大熊猫栖息地利用格局的影响进行了分析.研究结果表明:(1)地震前5年,大熊猫对栖息地的利用格局在年间没有显著变化;(2)地震后的2年间,大熊猫对栖息地的利用格局在年间没有显著变化;(3)地震前后连续7年调查的样线中,大熊猫对栖息地的利用格局在年间没有显著变化;(4)在龙溪一虹口保护区,大熊猫对样线的利用与滑坡体的面积没有显著关系,即大熊猫对栖息地的空间利用没有受到滑坡体面积大小的显著影响;(5)虽然从现有资料发现地震对大熊猫栖息地的利用格局没有显著的影响,但仍然需要在震后加强对大熊猫及其栖息地的保护.     

四川邛崃山系小熊猫分布

2000-2002年,对四川邛崃山系11个县(市)的小熊猫分布状况进行了调查.共完成调杏样线2332条,发现有小熊猫痕迹样线130条,分布在9个县,样线平均遇见率为5.57%,样线遇见率最高的是汶川县和崇州市;估算整个山系有小熊猫栖息地面积5107.82km2,以宝兴、汶川和天全3县的面积最大,分别占总面积的34.08%、30.65%和16.61%,这3个县占总面积的81.34%;小熊猫分布密度等级最高的区域是崇州市的四川鞍子河自然保护区.在山系内7个自然保护区中有小熊猫栖息地面积2166.84km2,占栖息地总面积的42.4%.小熊猫空间分布格局极不连续,存在许多空白区域,栖息地存在破碎化趋势,应进一步加强对邛崃山世界遗产地内小熊猫种群的保护.     

Parasite Threat to Panda Conservation

The giant panda is a global symbol of wildlife conservation that is threatened by historic and current habitat loss. Despite a great deal of research on the physiology, reproductive biology, and diet of pandas in the wild and in captivity, there is little information on wild panda mortality. Here we integrate previously unavailable data on the mortality of wild pandas. We report on three recent phases of panda mortality: deaths due to bamboo flowering in the 1970s and 1980s, surprisingly extensive poaching in the 1980s and 1990s, and a parasitic infection over the past few years. Our analyses suggest that the current most significant threat to wild panda survival is disease due to extraintestinal migration (visceral larval migrans) by an ascarid nematode. We demonstrate that the probability of death of wild pandas being caused by this disease increased significantly between 1971 and 2005 and discuss the possible factors leading to the emergence of this disease. Electronic Supplementary Material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

洞庭湖流域麋鹿等哺乳动物濒危灭绝原因的分析及其对麋鹿重引入的启示

洞庭湖流域曾有亚洲象(Elephasmaximus)、犀(Rhinocerossp.)、麋鹿(Elaphurusdavidianus)、川金丝猴(Rhinopithecusroxellana)、长臂猿(Hylobatessp.)、大熊猫(Ailuropodamelanoleuca)、梅花鹿(Cervusnippon)、棕熊(Ursusarctos)等哺乳动物分布,但受古气候、古地理以及人类活动的影响,这些哺乳动物已在洞庭湖流域灭绝。这些哺乳动物的濒危和灭绝既受自然环境变化和灾变的影响,也与物种本身生物学特性和人类活动有关,尤其与人类捕杀和生境丧失有关。据古籍记载分析:在洞庭湖流域,亚洲象和犀于北宋末期灭绝或已南迁,而野生麋鹿、大熊猫、川金丝猴、长臂猿、梅花鹿和棕熊等于19世纪末灭绝。根据我们对30个自然保护区或森林公园野生动物资源实地调查的结果,在洞庭湖流域已记录到21种国家重点保护哺乳动物,其中有5、6、10种哺乳动物分别处于“极危”、“濒危”、“易危”等级,这表明物种濒危的过程仍在继续。导致这些现生哺乳动物濒危的主要原因是生境丧失、人类猎捕、环境污染等,而人类活动干扰对现生濒危物种存活的影响越来越大。洞庭湖流域重引入麋鹿需采取人类协助生存策略:提供足够的且受洪水影响小的适宜生境、保证稳定的奠基者种群数量、减少人为干扰、调控种群密度、实施社区共管和生计替代项目、加强疾病防治、完善保护措施、加大保护基金投入、加强生境监测和湿地恢复等。     

大熊猫初生胎儿及幼仔组织器官的病理学研究

To elucidate the cause of prenatal and early postnatal death in giant panda, pathological studies have been carried out on paraffin-fixed tissue sections from two fetuses and four cubs. The fetuses appeared to have classical atrophic changes in lung and hemorrhage in multiple organs, whereas the cubs showed purulent inflmnmation in various organs, most profound in lung and umbilicus. Localized infusion of bacteria and neutrophils were identified in the focus. Acute enteritis and hepatitis were also observed, as well as purulent encephalitis in one case. Variable degrees of congestion, hemorrhage, denaturalization and putrescence were evident in heart, liver, spleen, kidney, intestines, and lymph nodes. The results indicated that the fetuses died from suffocation whereas the cubs died from infection.     

圈养小熊猫繁殖行为变化及繁殖行为对策

In order to investigate the change of reproductive behaviors and understand reproductive strategies of both male and female red pandas, one-year behavioral observation was conducted through the focal sampling method in the Chengdu Research Base of Giant Panda Breeding from December 1999 to November 2000. Our results indicated that reproductive behaviors showed significant differences between the estrous and non-estrous seasons. Frequencies of the rubbing anogenital, sniffing and licking marking were much higher in estrus than in non-estrus. Bleating only appeared in the estrus and can be regarded as an estrous indicator.The result also demonstrated that both male and females applied different reproductive behavioral strategies. Frequencies of activity, rubbing anogenital, licking and sniffing marking, and bleat were much higher in the male than in the female. However, those of resting and investigating were much lower in the male than in the female. This indicated that the male was more active than the female during the estrus and might imply that the male acts mainly as an estrous message sender, and the female as a message receiver in the estrous season.     

小相岭大熊栖息地干扰调查

强烈的人类干扰是大熊猫濒危的主要原因。本于2001年6～7月对四川省小相岭大熊猫栖息地内竹子开花、火灾、滑坡3种自然干扰类型和采伐、放牧、采药、公路、偷猎、割竹采笋、耕作、开矿和旅游9种人类干扰类型进行了调查。调查样方878个,干扰发生在海拔1800～4000m。结果表明：自然干扰都不严重,干扰比例最高的竹子开花,也仅占1．14％。采伐、放牧、采药和公路是主要人类干扰类型,分别占调查总样方的34．9％,26．3％,5．1％和4．2％。干扰最严重的地段是海拔3000～3499m,采伐样方的49．67％,放牧样方的47．19％,采药样方的31．11％均发生在该段,同时有48．89％的大熊猫活动痕迹样方也在该海拔。近年来,由于国家实施“天然林保护工程”,采伐已不再是主要干扰,而放牧、采药和偷猎等活动在短时间内仍不会减少。     

佛坪自然保护区野生大熊猫交配行为的观察

On March 26 and 27, 2003, we observed six and five giant pandas assembled together respectively in Huodiba and Lijiagou districts of Sanguanmiao, Foping Nature Reserve, Shaanxi Province, China. The mating sites were located in the coniferous forest and mixed coniferous and deciduous forest, with steep slope and sparse shrubs and bamboos. Wind power was estimated over 3rd category in those two days. The competition and lighting among male pandas were observed, and the mating right with the female panda was mainly based on the hierarchical ordering. However, not all the winners during competition can be access to mating with the female panda. For wild giant panda, its mating system is maybe plastic, which perhaps is affected by environment, time and panda population itself. Cubs or sub-adults are observed oceurring at the mating site, which is considered to be linked with the learning of the reproductive behavior. Our results maybe provide a useful guideline to the management and breeding of giant pandas in the captivity.     

大熊猫和小熊猫粪便DNA提取的简易方法

采集了大熊猫和小熊猫的新鲜粪便样品 ,使用 1 0 0 %乙醇保存。通过重复离心富集研究动物的肠道脱落细胞 ,并使用乙醇和双蒸水洗涤以除去抑制物。用 1 %的SDS快速裂解细胞 ,离心除去残渣后 ,向裂解液中加入蛋白酶进行消化。消化结束后使用等体积的酚 /氯仿抽提 ,乙醇沉淀DNA。用双蒸水溶解粪便DNA后 ,使用PCR产物纯化试剂盒对粪便DNA进行纯化。电泳检测结果显示 ,从乙醇保存的大、小熊猫粪便样品中抽提到高质量的粪便DNA。对线粒体控制区、细胞色素b基因、 1 2SrRNA基因的PCR扩增反应以及测序结果也证实了样品保存方法和DNA抽提方法可靠而高效。此方法使用实验室内常用的分子生物学试剂 ,不仅克服了分子粪便学研究中常见的抑制物粪便DNA微量降解严重等障碍 ,与商业化的粪便抽提试剂盒 (QIAampDNAStoolMiniKit,Qiagen)相比还是一种经济的试验方法 (抽提反应成本为试剂盒的 1 / 5 )。文中还对粪便DNA内细菌基因组等背景DNA可能对分子粪便学试验结果的影响进行了探讨。在基于PCR技术的遗传学研究中 ,对于植食性动物而言 ,粪便内的背景DNA对目标动物DNA片断的扩增和序列测定未见影响 ;但对于肉食性动物 ,则必须考虑被捕食者基因组对试验可能产生的影响 ,应谨慎对待