圈养东北虎血液生理参数的测定

东北虎(Panthera tigris altaica)是一种淑临灭绝的食肉动物,现已被列入IUCN濒危物种红色名录(Goodrich et al.,2015)。东北虎主要分布于俄罗斯远东地区、中国东北地区和朝鲜的北部(Kitchener et al.,2017)。根据中俄边境地区的野生东北虎数量调查统计,该地区生存着成年老虎39只和幼虎22只(Matiukhina et al., 2016)。     

半散养东北虎繁殖期PAE编码行为谱的构建

2006—2007年,采用全事件记录法、目标动物取样法和行为取样法对泰山东北虎园半散养东北虎繁殖期的各种行为进行了观察。基于以"姿势-动作-环境"(posture-act-environment,PAE)为轴心,以行为生态功能为依据的PAE编码系统,对半散养东北虎的行为进行分类和系统编码并构建PAE行为谱。研究结果区分和定义了14种姿势、47种动作和98种行为型;并将所区分、命名和编码的所有行为型归类于摄食、排遗、调温、发情、交配、分娩、冲突、育幼、通讯、聚群、休息、运动和杂类等13类;由此构建了基于PAE编码系统的半散养东北虎首个繁殖期行为谱。     

Ability of thawed tiger (Panthera tigris) spermatozoa to fertilize conspecific eggs and bind and penetrate domestic cat eggs in vitro.

Electroejaculates from tigers were collected and half was used fresh to inseminate tiger eggs in vitro and domestic cat eggs stored in a hypertonic salt solution. The remainder was pelleted, frozen in a solution of 20% egg yolk, 11% lactose and 4% glycerol, thawed and cultured with tiger and domestic cat eggs. The motility index ((sperm % motility)+(status rating x 20))/2 for thawed spermatozoa was about 86% of that in fresh aliquots. Of the 49 tiger oocytes inseminated in vitro with fresh spermatozoa, 34 (69.4%) cleaved, compared with 33 of 47 oocytes (70.2%) cultured with thawed spermatozoa (P > 0.05). Embryos generated by either sperm treatment could develop in vitro to the 16-cell or morula stage. Fresh and thawed tiger spermatozoa were equally capable (P > 0.05) of binding and penetrating the outer and inner zona pellucida of domestic cat eggs. These results demonstrate the ability of frozen-thawed tiger spermatozoa to (i) penetrate homologous and heterologous eggs and (ii) result in conspecific, advanced development of preimplantation embryos in vitro.     

Ultrastructure of oocyte maturation,fertilization, and early embryo development in vitro in the Siberian tiger (Panthera tigris altaica)

The application of assisted reproduction techniques to wild cats has been stalled by a lack of basic knowledge of the reproductive biology in these species. In this study, the ultrastructure of Siberian tiger (Panthera tigris altaica) cumulus-oocyte-complexes (COCs), as well as in vitro produced (IVP) zygotes and embryos were investigated, to estimate the normality of the manipulated reproduction processes. Adult female tigers were subjected to a purified porcine pFSH/pLH stimulation treatment followed by oocyte aspiration. According to morphological appearance at the stereomicroscopical level, COCs were classified as mature, immature, or degenerated, and then allocated into the following groups: presumptively immature COCs, which were in vitro matured (IVM-group) before fixation; presumptively mature COCs, which were either fixed after retrieval (pre-IVF-group), following in vitro insemination (IVF-group) or following in vitro insemination and subsequent in vitro culture (IVC-group). All specimens were processed for light and transmission electron microscopy (TEM). Both the IVM- and pre-IVF-group included oocytes in meiotic stages ranging from prophase I to metaphase II, and some prophase I oocytes in the IVM-group were apparently in their growth phase. The IVF-group presented features of presumptive normal fertilization, but aberrations such as polynucleation were also noted. The IVC-group included cleavage stage embryos of which, however, many were polynucleated. In conclusion, the procedures used for stimulation, aspiration, and classification of COCs resulted in retrieval of a heterogeneous population of oocytes which, following IVF and IVC, displayed a high rate of developmental deviations.     

Twenty-one novel tri- and tetranucleotide microsatellite loci for the Amur tiger (Panthera tigris altaica)

Amur tiger is the largest subspecies of tiger in the world and his conservation has also received much attention. In this study, we isolated and characterized twenty-one tri- and tetranucleotide microsatellite markers from this species. The number of alleles for each locus ranged from two to nine in a group of 60 individuals and the observed and expected heterozygosities were 0.333–0.917 and 0.302–0.822, respectively. The overall discrimination power and exclusion probabilities in parentage and paternity testing for these markers were 1.00, 0.9947 and 0.9999, respectively, indicating high-resolution power of microsatellite markers.     

Coprology of Panthera tigris altaica and Felis bengalensis euptilurus from the Russian Far East

Fecal samples from the Siberian tiger (Panthera tigris altaica) and the Amur cat (Felis bengalensis euptilurus) from Far Eastern Russia, were examined for parasites. A natural sedimentation methodology was used and a complete examination of all the sediment was performed. This fecal investigation allowed us to isolate and identify several developmental stages of gastrointestinal, hepatic, and respiratory parasites. Five parasites were found from P. t. altaica: 11 trematodes (Platynosomum fastosum) and 4 nematodes (Strongyloides sp., Ancylostomatidae, Toxascaris leonina, and Toxocara cati). Five parasites were found from F. b. euptilurus: 1 cestode (Diplopylidium sp.) and 4 nematodes (Trichuris sp., Ancylostomatidae, Toxascaris leonina, and Aelurostrongylus abstrusus). In addition, trophozoites of the amoeba Acanthamoeba sp. were detected in tiger feces.     

东北虎幼体消化系统蛋白水解酶的初步研究

蛋白水解酶在许多生命活动中是必需的物质(Vassalli and Pepper,1994)。蛋白质的酶解修饰(Xuet al.,1999)、细胞迁移、组织再生与修复、消化系统对蛋白质的消化等均与蛋白水解酶有关(Baimbridgeet al.,1992),且蛋白水解酶功能失调会导致许多疾病(Teichertet al.,1989)。东北虎(     

笼养和半散放东北虎行为时间分配及活动规律差异的比较

1998年 3月～ 2 0 0 1年 4月 ,在哈尔滨动物园和黑龙江东北虎林园采用全事件取样法、扫描取样法和目标取样法 ,对笼养和半散放东北虎的行为时间分配和活动规律进行了比较研究。结果表明 ,在时间分配方面 ,笼养东北虎的休息远远高于半散放东北虎 ,而摄食、站立和其它行为则明显低于半散放东北虎 ,笼养和半散放东北虎的运动较为接近。在活动规律方面 ,虽然笼养和半散放东北虎均有 2个运动的高峰期 ,但笼养东北虎的相对集中且变化幅度较大 ;半散放东北虎的休息有 1个明显的高峰期和 2个不明显的高峰期 ,与笼养东北虎的 1个高峰期有很大不同 ;笼养东北虎的摄食仅出现在 16∶0 0～ 17∶0 0 ,而半散放东北虎则在 1d中均有发生 ,且在 9∶0 0～ 11∶0 0和 14∶0 0～ 16∶0 0有 2个高峰期 ;笼养和半散放东北虎的站立和其它行为变化规律较为相似 ,但半散放东北虎的发生强度要远远高于笼养东北虎。通过对笼养和半散放东北虎各种行为进行的配对T检验 ,发现二者的休息、摄食、站立和其它行为存在极显著差异 (P<0 0 0 1)。     

黑龙江省东北虎分布区域及其数量动态

2005 年1月至2006年12月利用划分监测区的方法对黑龙江省境内野生东北虎进行监测,通过测量前足迹掌垫宽度的方法对野生东北虎进行个体识别,确定了野生东北虎的分布区及其种群变化情况.结果表明,黑龙江省有野生东北虎12～14只.其中,完达山东部分布区种群数量为5～6只,由2只雄性个体,2只雌性个体和1~2只亚成体组成;老爷岭南部分布区种群数量为3～4只,由1~2只雄性个体,1只雌性个体和1只亚成体组成;老爷岭北部分布区种群数量为4只,由2只雄性个体和2只雌性个体组成;张广才岭南部分布区监测期间未发现东北虎活动信息.监测结果与近几年调查的结果相比,分布区域和种群数量都发生了变化.在4个分布区中,老爷岭北部分布区是再次出现东北虎活动的区域,张广才岭南部分布区未监测到东北虎的活动信息,可能没有分布.完达山东部分布区、老爷岭南部分布区和老爷岭北部分布区种群数量略有上升,张广才岭南部分布区呈现下降趋势.     

半散养状态下东北虎交配行为的观察

在2006～2007年,采用行为取样法对泰山东北虎园7只(3雌,4雄)半散养状态下(园区占地面积20 000 m2)的东北虎的交配活动进行了为期22 d的观察,旨在了解无人干扰下东北虎的交配过程及其交配模式.结果显示:东北虎在交配过程中无锁结现象,雄虎通常在一次爬跨多次抽动后即出现射精.在1 h内出现2次射精次数的比例占总射精次数的74.0%.东北虎的交配模式属于Dewsbury分类系统中的第11种,即无锁结、有抽动、单次插入、多次射精.交配期内,每只雌虎平均日邀配28±3次,平均邀配持续时间为20.0±1.2 s;每只雄虎平均日爬跨14±1次.平均抽动持续时间为20.9±0.5 s,平均交配持续时间为45.2±1.3 s.     

In-vitro induction of capacitation of fresh and frozen spermatozoa of the Siberian tiger (Panthera tigris)

Electroejaculates from 5 tigers were split and half of each was assayed fresh while the remainder was frozen and thawed before being assayed. Preincubation time, temperature and removal of seminal plasma were evaluated for their effect on in-vitro capacitation. Ability of spermatozoa to penetrate oocytes, as measured by the zona-free hamster egg-sperm penetration assay (SPA), was used as verification of capacitation. Results of the experiments with fresh semen indicate that: (1) preincubation time affects the fertilizability of tiger spermatozoa with 2 h appearing optimal, (2) a preincubation temperature of 37 degrees C results in significantly higher penetration rates than does a 22 degrees C treatment, and (3) tiger seminal plasma does not appear to contain decapacitation factors, as has been reported for several other species. Frozen semen experiments indicate that (1) frozen-thawed tiger spermatozoa must be removed from the environment of the semen extender before capacitation can take place, and (2) the freeze-thaw procedure results in a shortening of the required capacitation time.     

Characteristics of fresh and cryopreserved Siberian tiger (Panthera tigris altaica) semen and its seminal plasma chemical constituents

Semen was collected by electroejaculation from four Siberian tigers. The cells were separated from the seminal plasma by centrifugation, extended in TEST-yolk buffer with 4% glycerol and frozen for a Sephadex filtration assay. The seminal plasma was analyzed for chemical constituents and characteristics. There were significant differences between tigers for volume, cell concentration, pre- and post-thaw motility, total cells, percent live/normal cells, pH, specific gravity, albumin, alkaline phosphate, alanine transaminase, aspartate transaminase, total protein, urea nitrogen, potassium, chloride, carbon dioxide and glucose. There were no differences between tigers for total live cells, osmotic pressure, acid phosphatase, sodium, phosphorus, calcium, or creatinine. There were significant differences between months only for pre-freeze motility, albumin, chloride, carbon dioxide, and calcium. The differences in seminal plasma chemistry may be useful for semen quality evaluation after successful breeding techniques have been developed. As the data base of information concerning the reproductive physiology of tigers increases, application of advanced technologies may become a reality.     

半散放东北虎行为活动时间的分配

1998年5年－2001年4月在黑龙江东北虎林园,采用焦点动物取样法对10只半散放东北虎（5雌5雄）全年的昼间行为活动时间分配进行了研究。结果表明：（1）半散放东北虎的运动（37．53％）和休息（32．45％）行为较多,站立（6．24％）,摄食（10．13％）,社会（8．36％）和其他（5．29％,包括饮水,排遗,修饰等）行为较少;（2）不同季节的活动规律相似,运动,摄食,站立和社会行为在上午和下午各有1个高峰期,休息在中午有1个高峰期,运动和行为在存,冬季明显比春,秋季增多。将上述结果与笼养和野生东北虎的进行了比较,并提出了在今后的野化训练中应采取的措施。     

饲养条件下东北虎交配行为的观察

1999年和 2 0 0 0年的 1 1～ 1 2月期间共记录了黑龙江东北虎林园 1 6对东北虎的 1 693次交配事件 ,对其中 1 1 92次成功的交配事件进行了分析 ,建立了东北虎的交配行为谱 ,记录了 1 6种个体行为和 7种社会行为。东北虎的交配持续期为 (6 5 0± 2 5 0 )d ,不同繁殖对的交配次数差异很大 ,最少为 49次 ,最多可达 1 1 3次 ,交配成功率为 70 41 %。东北虎的交配高峰出现在 8∶0 0～ 1 0∶0 0时和 1 6∶0 0～ 1 8∶0 0时。交配成功的次数在第 3～ 5d达到高峰 ,失败的次数在开始交配时最高 ,以后则逐渐降低。在将雌、雄虎赶入小围栏内的首次爬跨前东北虎需花费 (2 0 2 4± 1 2 85 )min的时间做准备 ,其后的各次爬跨与首次相似 ,只是花费时间大大缩短 ,且持续的时间相差很大 ,最短仅为 4 70s,最长则达 6 3 1min。交配后的休息过程同时也是下一次交配前的准备阶段。每次将雌、雄虎赶入小围栏内东北虎要连续进行 (9 40± 4 60 )次爬跨 ,整个交配过程持续 (1 2 67± 3 0 5 )min ,每次爬跨历时 (1 0 0 9± 5 45 )s。     

东北虎源等孢球虫的发现

对东北虎粪便进行虫卵检查,分离获得了自然感染的球虫卵囊。对球虫卵囊的形态和经培养发育后的各阶段的卵囊形态进行了观察。东北虎源球虫卵囊符合等孢属球虫的形态学特征。根据Genbank 上发表的猫等孢球虫18S rDNA 序列(L74671)设计一对特异性引物,利用PCR 技术对东北虎源球虫的18S rDNA 进行了扩增。扩增得到一段360 bp 的DNA 片段,并对该片段进行了序列测定。测序结果和其它17 种原虫的相应序列进行序列同源性比较分析,分析显示东北虎源球虫和其它等孢属球虫单独聚为一类。结合对东北虎源球虫的卵囊形态、卵囊发育情况及序列分析结果,确定其为等孢属球虫。      

吉林珲春自然保护区野生东北虎捕食家畜的状况

2002 年5 月至2004 年10 月通过跟踪调查,我们搜集到野生东北虎捕食珲春自然保护区居民家畜的数据,共计38 起捕食事件,有51 头家畜遭到捕食。这些数据包括:捕食事件发现时间、被捕食家畜的数量、类型、年龄、性别、虎痕迹照片和捕食点的GPS 数据。运用Mann-Whitney U 检验对数据的差异性进行了检验;结合ArcView GIS和MapIn fo 分析,对捕食点的空间属性进行了量化,用多个生态因子对其进行了表征;运用PCA 分析了多因子中的主要成分。这些数据表明:在珲春,东北虎主要捕食牛和马,对猪和羊没有捕食,对牛和马的捕食也不存在偏好。东北虎倾向捕食成年个体,并且,大多数情况下只捕食1～2 个个体。成功的捕食事件中,多为1 头遭捕食;该捕食策略为虎权衡后的选择。捕食家畜事件多发生在3 ～11 月的非冰冻期。影响捕食地的生态变量重要性排序为:距最近居民区距离> 海拔> 距最近道路距离> 土地利用(Ⅱ型)类型> 坡度> 坡向> 距最近水源距离> 植被类型> 道路类型> 土地利用(I 型)类型。距最近居民区距离为:3 666 ± 2 308 m,极显著地近于休息卧迹、警戒卧迹和刨痕。该距离更多地集中在4 000 m以下区域。海拔为280 ± 114 m,极显著地低于休息卧迹,显著低于警戒卧迹和刨痕。由于当地居民区多集中在海拔较低的区域,因此,这种显著的不同可以看成是一个伴生的结果。距最近道路距离为:522 ± 543 m,极显著地近于刨痕,显著近于休息卧迹和警戒卧迹。上述2 个人为干扰因子,对捕食的影响具有相似性。对上述3 个生态因子的详细研究说明,虎并不愿意有意识地如此接近人类聚居区,而是家畜食物的诱惑,令虎冒险前往。在26 个捕食点中,14 个在用材林,6 个在特护林,3 个在农田,2 个在灌木林,1个在居民区。被调查的当地居民共损失了 11 851 kg家畜,合人民币116 126元。     

基于最小费用距离模型的东北虎核心栖息地确定与空缺分析

东北虎(Panthera tigris altaica)是世界珍稀濒危物种之一。近年来,栖息地破碎化、猎物密度降低和人类的猎杀使得野生东北虎的数量急剧下降。在中国,野生东北虎仅分布于黑龙江省和吉林省东部山区,数量不到20只。本文以最小费用距离模型作为系统框架,综合植被类型、地形、人类影响等因素,确定了黑龙江东部山区的东北虎核心栖息地分布,并通过与现有保护区分布的比较,提出目前东北虎的保护空缺。结果表明,东北虎潜在核心栖息地总面积9576.37km2,占研究区域总面积的12.36%,主要分布在老爷岭东部和北部、完达山东北部、张广才岭以南、太平岭北部几个区域,然而,潜在核心栖息地完全没有被保护,就东北虎的长期生存与进化而言,在研究区域内建立合理有效的保护区网络具有重大意义。     

mtDNA indicates profound population structure in Indian tiger (Panthera tigris tigris)

We analyzed mtDNA polymorphisms (parts of control region, ND5, ND2, Cytb, 12S, together 902 bp) in 59 scat and 18 tissue samples from 13 Indian populations of the critically endangered Indian tiger (Panthera tigris tigris), along with zoo animals as reference. Northern tiger populations exhibit two unique haplotypes suggesting genetic isolation. Western populations from Sariska (extinct in 2004) and Ranthambore are genetically similar, such that Ranthambore could serve as a source for reintroduction in Sariska. Zoo populations maintain mitochondrial lineages that are rare or absent in the wild.