从头组装豚鹿血液转录组揭示雌雄基因表达差异

豚鹿Axis porcinus是国家一级重点保护野生动物,为理解其基因表达上的基本特征,本研究采用RNA-seq技术对16个豚鹿血液样本进行了测序和从头组装,一共得到615 548条unigenes,Contig N50为803 bp。将组装的转录本进行同源序列比对注释,发现豚鹿血液在信号转导、抗病毒感染和免疫系统等通路上表达的基因最丰富。通过比较雌雄豚鹿的差异表达基因（DEGs）,鉴定了1 781个在雄性中显著上调和14个显著下调的性别差异表达基因,雄性的高表达DEGs远多于雌性,且主要富集在精子形成、睾丸功能和自身免疫相关通路,暗示雌雄豚鹿在生殖发育和自身免疫方面存在较大差异。部分可能与鹿角生长相关的基因（Rela、ADAMTS2、OLIG1、Snai1、PTH1R、TGFBI、MMP9）均在雄性豚鹿中显著高表达。本研究为丰富豚鹿转录表达提供了基础数据,为进一步深入挖掘豚鹿功能基因、分子标记开发以及鹿角形成机制提供了参考。     

白鱀豚某些血液生化指标的测定

关于海豚和几种淡水豚类的血液学和血液生化指标国外有过研究4-13。白鱀豚(Lipotes vexillifer)血液有形成份已有过报道2,3。本实验对白鱀豚血液生化指标进行了研究,以期建立白鱀豚血液正常生理生化指标,为白鱀豚的临床诊断,健康监测和保健措施提供血液学参数,并且为完善淡水豚类血液学比较研究提供基础资料。       

笼养丹顶鹤成体与亚成体血液生理生化指标比较

为了揭示丹顶鹤(Grus japonensis)不同年龄和性别个体之间血液生理生化指标的差异性,对沈阳动物园42只笼养丹顶鹤(成体24只,亚成体18只)的15项血液生化指标和13项血液生理指标进行了比较。所测得的血液生理生化指标在不同性别之间不存在显著差异(P0.05)。血清肌酐成体(37.05±17.03)mmol/L、亚成体(22.57±11.68)mmol/L,血清尿酸成体(878.22±514.41)mmol/L、亚成体(426.17±207.74)mmol/L,血清甘油三酯成体(7.73±4.75)mmol/L、亚成体(3.51±4.07)mmol/L,这3项指标均为成体高于亚成体,差异性显著(P0.05)。这几项指标的变化表明,丹顶鹤在成体和亚成体时期采取了相同或不同的生态适应策略。     

白暨豚某些血液生化指标的测定

关于海豚和几种淡水豚类的血液学和血液生化指标国外有过研究。白暨豚(Lipotes vexillifer)血液有形成份已有过报道。本实验对白暨豚血液生化指标进行了研究,以期建立白暨豚血液正常生理生化指标,为白暨豚的临床诊断,健康监测和保健措施提供血液学参数,并且为完善淡水豚类血液学比较研究提供基础资料。     

扎龙自然保护区雌性丹顶鹤血液生化指标

2004年11月至2005年9月,采用自动生化分析仪对扎龙自然保护区10只雌性成体丹顶鹤在越冬期、繁殖期和迁徙期的20项血液生化指标（总蛋白、白蛋白、球蛋白、直接胆红素、间接胆红素、总胆红素、血糖、尿素氮、肌酐、总胆固醇、甘油三酯、谷草转氨酶、谷丙转氨酶、碱性磷酸酶、肌酸肌酶、羟丁酸脱氢酶、乳酸脱氢酶、钙离子、无机磷和镁离子）进行了分析.结果表明：除总蛋白、球蛋白和血糖外,研究区雌性丹顶鹤血液中的其他指标均在不同时期之间表现出显著（P<0.05）或极显著差异（P<0.01).说明总蛋白、球蛋白和血糖可作为雌性丹顶鹤血液生化指标的可靠参考值,其他17项血液生化指标的年变化动态体现出雌性丹顶鹤在不同生理时期的生理特征和生态适应性.如果以这17项血液生化指标作为雌性丹顶鹤血液生化指标的参考值,就必须考虑不同生理时期对其含量的影响.     

豚鹿MHC-DOA基因克隆及其外显子2遗传多样性研究

主要组织相容性复合物(MHC)在脊椎动物的免疫系统中起着重要的作用,MHC-DOA基因是MHCⅡ类的非经典基因,其外显子2多态性丰富。豚鹿Axis porcinus是我国极度濒危的动物。本研究以成都动物园圈养的38头豚鹿为研究对象,提取豚鹿血液总RNA,反转录合成cDNA为模板,利用PCR方法克隆到豚鹿MHC-DOA序列;同时利用PCR方法扩增38头豚鹿的MHC-DOA基因外显子2基因并测序,再进行多态性分析。结果显示:豚鹿MHC-DOA基因开放阅读框长753 bp,编码250个氨基酸残基;蛋白质同源性分析表明,豚鹿MHC-DOA基因与东欧马鹿Cervus elaphus hippelaphus的同源性最高(98.4%);38头豚鹿的MHC-DOA外显子2共有10种单倍型,整体遗传多样性水平中等;MHC-DOA外显子2部分序列中发生了核苷酸的缺失和插入,进而引起DOA蛋白序列发生改变。豚鹿MHC-DOA多态性的研究对豚鹿种群遗传结构调查、遗传资源的保护具有重要意义。     

豚鹿几种激素的季节性变化模式研究

动物季节性发情繁殖涉及下丘脑-垂体-性腺轴系统复杂的神经内分泌过程。豚鹿Axis porcinus的繁殖行为在一定程度上受季节影响。本研究通过检测2015年5月、8月和10月雌性豚鹿血液中繁殖相关激素含量的变化模式,结合温度、湿度和光照时间的变化情况,粗略描绘了春季、夏季和秋季豚鹿内分泌调节动态。结果发现,豚鹿属于短光照繁殖类型动物,其繁殖激素存在季节性的周期变化。8月温湿度指数较高且光照时间较长,豚鹿血浆中三碘甲腺原氨酸、四碘甲腺原氨酸含量极显著低于5月、10月(P0.01);促黄体素、催乳素含量极显著低于5月、10月(P0.01);外周血液促卵泡素含量极显著低于5月(P0.01);夏季高温、高湿和长光照影响了豚鹿繁殖相关激素的分泌。     

白暨豚血液有形成分

豚类血液学的研究,自1939年诺尔(Knoll)广泛研究了鲸类胚胎的血液以来,1979年德蒙蒂(De monte)和皮莱里(Pilleri)比较了21种海豚和几种淡水豚的血液学资料。关于白暨豚(Lipotes vexillifer)的血液资料,除刘仁俊(1982)为白暨豚“淇淇”治疗外伤时,进行过白细胞分类计数外,至今未见详细的研究报道。为建立白暨豚血液生理指标和为疾病诊断提供可靠的依据,本文对白暨豚外周血液的有形成分进行了分析研究。     

基于微卫星位点的人工圈养豚鹿亲子关系鉴定

豚鹿属我国国家I级重点保护动物,目前国内野生种群已经灭绝,人工圈养数量少,已极度濒危。成都动物园是国内最大的豚鹿饲养单位,对该园内豚鹿进行亲子鉴定及遗传谱系建立是我国豚鹿拯救工程中一重要环节。本文利用7个微卫星标记对成都动物园27只豚鹿个体进行了基因分型,在母本已知情况下成功鉴定了13对父子关系,其中排除法鉴定8对,似然法鉴定5对且置信度达95%。将亲子鉴定的结果辅以动物园豚鹿圈养的历史记录,我们构建了该园豚鹿的遗传谱系图。本文的研究成果将为后续人工繁殖中亲本雌雄配对的个体选择以及种群的遗传管理提供参考。     

圈养婆罗洲猩猩部分血液参数值测定

采集6家动物园18只健康婆罗洲猩猩（Pongo pygmaeus）的24份血液样本,进行血液常规及生化参数检测。依据性别（雄性14份,雌性10份）和年龄（幼龄个体7份,亚成体6份,成体11份）对研究对象进行分组,分别统计不同组别样本的血液参数值,并进行组间比较。研究结果表明,雌、雄两组之间血液参数没有显著差异（One Way ANOVA test,P > 0.05）。淋巴细胞（LYM）比率随着年龄的增长而降低（One Way ANOVA test,F = 5.200,P < 0.05）。雄性成年个体血液中的谷丙转氨酶（ALT）浓度显著高于雌性个体（One Way ANOVA test,F = 4.911,P < 0.05）,其他参数未呈现性别间差异（One Way ANOVA test,P > 0.05）。总蛋白（TP）呈现年龄组间的差异（One Way ANOVA test,F = 4.715,P < 0.05）。所检测圈养婆罗洲猩猩血液中葡萄糖含量较高,应在饲养过程中给予关注。     

圈养毛冠鹿血液生理生化指标测定

本文采集了8只不同年龄段毛冠鹿的14个血液样品,并检测和分析了样品中的血细胞、血蛋白、电解质、生化酶类等44项指标的含量。初步建立了圈养毛冠鹿的血液学生理生化指标参考值,为今后加强毛冠鹿的人工饲养及繁育研究提供了参考依据。     

Widespread hybridization in the introduced hog deer population of Victoria,Australia, and its implications for conservation

In Australia, many species have been introduced that have since undergone drastic declines in their native range. One species of note is the hog deer (Axis porcinus) which was introduced in the 1860s to Victoria, Australia, and has since become endangered in its native range throughout South‐East Asia. There is increased interest in using non‐native populations as a source for genetic rescue; however, considerations need to be made of the genetic suitability of the non‐native population. Three mitochondrial markers and two nuclear markers were sequenced to assess the genetic variation of the Victorian population of hog deer, which identified that the Victorian population has hybrid origins with the closely related chital (Axis axis), a species that is no longer present in the wild in Victoria. In addition, the mitochondrial D‐loop region within the Victorian hog deer is monomorphic, demonstrating that mitochondrial genetic diversity is very low within this population. This study is the first to report of long‐term persistence of hog deer and chital hybrids in a wild setting, and the continual survival of this population suggests that hybrids of these two species are fertile. Despite the newly discovered hybrid status in Victorian hog deer, this population may still be beneficial for future translocations within the native range. However, more in‐depth analysis of genetic diversity within the Victorian hog deer population and investigation of hybridization rates within the native range are necessary before translocations are attempted.     

Secretory ribonuclease genes and pseudogenes in true ruminants

Mammalian pancreatic ribonucleases (RNase) form a family of extensively studied homologous proteins. Phylogenetic analyses, based on the primary structures of these enzymes, indicated that the presence of three homologous enzymes (pancreatic, seminal and brain ribonucleases) in the bovine species is due to gene duplication events, which occurred during the evolution of ancestral ruminants. In this paper the sequences are reported of the coding regions of the orthologues of the three bovine secretory ribonucleases in hog deer and roe deer, two deer species belonging to two different subfamilies of the family Cervidae. The sequences of the 3′ untranslated regions of the three different secretory RNase genes of these two deer species and giraffe are also presented. Comparison of these and previously determined sequences of ruminant ribonucleases showed that the brain-type enzymes of giraffe and these deer species exhibit variations in their C-terminal extensions. The seminal-type genes of giraffe, hog deer and roe deer show all the features of pseudogenes. Phylogenetic analyses, based on the complete coding regions and parts of the 3′ untranslated regions of the three different secretory ribonuclease genes of ox, sheep, giraffe and the two deer species, show that pancreatic, seminal- and brain-type RNases form three separate groups.     

豚鹿的饲养管理与繁殖

本文报道成都动物园豚鹿的饲养管理、疾病预防和繁殖。适宜的饲养环境、宽敞的兽舍、高纤维含量和多样化的食物是豚鹿正常生长发育、繁殖的重要因素。     

林麝血液生理生化指标研究

以健康雄麝、健康雌麝和患化脓病林麝各10头为研究对象,比较它们之间的血液生理生化指标差异。结果表明:(1)健康雄麝和健康雌麝的甘油三酯、极低密度脂蛋白存在显著差异(P<0.05)。雄麝的甘油三酯为0.32mmol/L,极低密度脂蛋白为0.14mmol/L;雌麝的为0.47mmol/L,0.21mmol/L;其他生理生化指标无显著差异。(2)健康林麝与患化脓病林麝的白细胞、淋巴细胞百分比、嗜中性粒细胞百分比存在极显著差异(P<0.01);健康林麝的白细胞数量为29.08×109个/L、淋巴细胞百分比为62.01%、嗜中性粒细胞百分比为34.05%;患化脓病林麝的为104.67×109个/L、50.87%、45.37%。健康林麝与患化脓病林麝之间白蛋白、球蛋白、天门冬氨酸氨基转移酶、乳酸脱氢酶、总胆固醇、肌酐、血糖存在显著差异(P<0.05),健康林麝的白蛋白为44.44g/L、球蛋白为33.6g/L、天门冬氨酸氨基转移酶为316.43U/L、乳酸脱氢酶为700.68U/L、总胆固醇为2.14mmol/L、肌酐为108.47μmol/L、血糖为7.29mmol/L;患化脓病林麝的为37.23g/L、47.45g/L、478U/L、840.75U/L、1.16mmol/L、85.52μmmol/L、5.04mmol/L。这些结果为养殖林麝化脓病的诊断提供了参考数据。     

人工育幼豚鹿幼仔的生长发育规律初步研究

测定了成都动物园圈养种群中9只人工育幼豚鹿幼仔(2004～2007年)体重和8只豚鹿的体尺,并观察了8只豚鹿的牙齿(0～63 d)的生长情况.结果为:幼仔在0～54 d内体长、肩高、后足长、耳长和尾长以及体重都随日龄呈直线增长趋势,其中体长和肩高与幼仔的日龄显著相关(P<0.01).初生幼仔齿式2(I 0/2, C 0/0, P 0/0),2周龄齿式2(I 0/2,C 0/0, P 2/2),7周龄齿式2(I 0/2,C 0/2或C 1/2, P 2/2).     

Facilitative interactions between an exotic mammal and native and exotic plants: hog deer (<Emphasis Type="Italic">Axis porcinus</Emphasis>) as seed dispersers in south-eastern Australia

Endozoochory by exotic mammalian herbivores could modify vegetation composition by facilitating the dispersal and establishment of exotic and native plant species. We examined the potential for endozoochoric dispersal of native and exotic plants by exotic hog deer (Axis porcinus) in south-eastern Australia. We quantified the germinable seed content of hog deer faecal pellets collected in five vegetation types within a 10,500-ha study area that was representative of their Australian range. Twenty exotic and 22 native species germinated from hog deer faecal pellets and significantly more native species germinated compared to exotic species. Seedlings of the encroaching native shrub Acacia longifolia var. sophorae emerged, but no native trees emerged and the percentage of grasses that germinated was low (11%). The species composition of germinants was similar among the five vegetation types. We estimated that the hog deer population in our study area could potentially disperse >130,000 viable seeds daily. Our study shows how an exotic mammal can disperse seeds from both native and invasive plants and highlights the need for endozoochory to be considered more widely in studies assessing the impacts of exotic mammals on plant communities.     

Dry season diets of sympatric ungulates in lowland Nepal: competition and facilitation in alluvial tall grasslands

Based on microhistological analyses of faecal material, we compared the early dry season diets of greater one-horned rhinoceros Rhinoceros unicornis, swamp deer Cervus duvauceli and hog deer Axis porcinus, which inhabit the same alluvial grassland habitat complex in lowland Nepal. Their diets were quite similar, both at the forage category level and within subcategories of graminoids and woody plants. Early successional tall grasses, especially Saccharum spontaneum, were the dominant food of all three species, underlining the key role of the threatened alluvial floodplains in large mammal conservation in South Asia. The two deer species ate significantly more graminoids (>66.5%) than did rhino (45.5%), and although they did not differ in proportions of graminoids, swamp deer ate significantly more late successional tall grasses (Narenga porphyrocoma and Themeda spp.) and short grasses (mainly Imperata cylindrica) than hog deer. The two deer consumed almost equal proportions of woody browse (ca. 10%), significantly less than that of rhino (33.0%). The prediction of the Jarman–Bell hypothesis, that large-bodied herbivores are less selective and subsist on lower quality graminoids than smaller ruminants, was not supported by the data. Based on this and previous studies in the same area we propose a conceptual model where the larger megaherbivores (rhino and elephant Elephas maximus) facilitate the smaller swamp deer and hog deer during the monsoonal growing season, while the smaller and more selective deer species outcompete the larger during the dry season when food is more limited. Owing to the all-year sprouting ability of S. spontaneum, facilitation may occur also in the dry season with low numbers of megaherbivores, thus accentuating competitive exclusion at higher deer densities.     

Predicting spacing behavior and mating systems of solitary cervids: a study of hog deer and Indian muntjac

This study investigates the validity of current theory for predicting ecological and allometric effects on space use, social structure and mating systems of poorly known solitary cervids, based on a comparative analysis of radio-telemetry data on hog deer Axis porcinus (N=32) and Indian muntjac Muntiacus muntjak (N=28). The larger and sexually size-dimorphic hog deer inhabit highly productive alluvial floodplains, where resource distribution is patchy and spatiotemporally unpredictable. As predicted for this species, site fidelity was low and range sizes varied among sex and age groups and among seasons. Hog deer were probably non-territorial, as home range sizes seemed too large to be exclusive when taking into account their high population density. Extensive movements of adult males during the rut implied "roaming" as a mating strategy. The smaller, forest-dwelling and sexually size-monomorphic muntjacs inhabit a more uniform and stable habitat. As predicted, muntjacs exhibited higher site fidelity than hog deer, and no seasonal variations in home range sizes. Adults exhibited relatively large home range overlap, both inter- and intrasexually. Hence, strict territoriality did not occur, but their well-defined home ranges and high site fidelity indicated some form of site-specific dominance. In conclusion, habitat characteristics were appropriate predictors of home range sizes and site fidelity. Body mass appeared to be a suitable predictor of intraspecific patterns in space use but a poor predictor of interspecific patterns, probably due to a confounding effect of habitat productivity.