大熊猫粪便类固醇激素研究进展

概述了大熊猫粪便样本采集和保存方法,系统总结了大熊猫粪便类固醇激素提取、测定方法和应用研究,对大熊猫粪便类固醇激素的应用进行了展望。     

粪样在大熊猫研究上的应用

大熊猫(Ailuropoda melanoleuca)数量少,传统取样法难以实施,样品的缺乏给大熊猫研究带来很大困难。而采集大熊猫粪样对大熊猫自身无损伤,粪样易获得和保存。本文总结了应用粪样进行的大熊猫食性与消化吸收、种群与个体识别、类固醇激素、寄生虫与病原微生物等方面的研究进展,对大熊猫粪样的应用前景进行了展望。     

粪便类固醇激素检测准确性的影响因素

近年来,采用非损伤性取样方法监测野生动物的生理状况越来越受到重视。本文对检测过程中影响动物粪便类固醇激素检测准确性的因素进行了分析总结,包括样品的新鲜程度、样品量和保存方法;激素代谢的日节律和季节性变化;动物的年龄、性别和繁殖状态等,以期为粪便类固醇激素检测技术在野生动物中的准确应用提供借鉴。     

川金丝猴尿液类固醇激素的保存时效性

尿液类固醇激素检测已经广泛运用于野生动物的生理机能、健康状况和繁殖状态的研究,由于分析条件以及样品采集时间限制,一般尿液样品需保存一段时间后才能在实验室分析,不同物种尿液类固醇在不同保存条件的保存时效有差异。本研究利用2只成年雄性和4只成年雌性的尿液样品共21份,确定川金丝猴尿液类固醇的保存时效和保存方法。结果表明:4℃时,雄性川金丝猴尿液睾酮、雌性尿液睾酮、孕酮的保存时效为7 d左右;雌性尿液雌二醇的保存时效在3 d以内。-20℃时,雄性川金丝猴尿液中睾酮的含量在60 d内保持稳定;雌性尿液中睾酮、孕酮和雌二醇的含量在60 d、45 d、60 d内保持稳定。-20℃保存川金丝猴尿液样品是一种有效、可靠的方法。     

大熊猫生殖生物学研究:Ⅰ.大熊猫发情期血清和尿液中促黄体素、孕酮和17β-雌二醇含量的变化

作者用放射免疫法和生物测定法对4只大熊猫发情期血清和尿液中的促黄体素、孕酮和17β-雌二醇的含量进行分析。结果表明:3只大熊猫在发情高峰期均出现雌二醇和促黄体素高峰;LH峰值出现在E_2峰值之后。根据雌兽在发情期血清和尿液中这二种激素含量的变化可为选择人工授精的最佳时间提供可靠的理论依据。     

HS-SPME-GC / MS 在大熊猫尿液化学成分检测中的应用

尿液在大熊猫化学通讯过程中具有重要作用。对大熊猫尿液中化学成分的检测是揭示大熊猫尿液中化学物质组成及其功能的关键。本实验通过使用顶空固相微萃取技术(Headspace-solid phase microextraction,HSSPME)对大熊猫尿液样品进行前期处理,继而利用气相色谱- 质谱联用技术(Gas chromatography-mass spectrometry,GC / MS)对大熊猫尿液中化学成分进行检测。共检测到56 个峰,通过在NIST (National Institute of Standards
and Technology)质谱库中进行检索,初步推定出其中的38 种物质。除此之外,还对HS - SPME 萃取头的净化方法进行了探索和改进。结果表明,顶空固相微萃取技术结合气相色谱- 质谱联用技术能够应用于大熊猫尿液中挥发性与半挥发性化合物的检测,并且能够得到较好的实验结果,为揭示大熊猫化学通讯机理提供基础。     

粪便类固醇激素研究简史及其在野生动物研究中的应用

系统回顾了粪便类固醇激素研究的历史,全面总结了国内外学者对动物粪便类固醇激素研究的现状,认为利用动物粪便研究类固醇激素具有完全无伤害性、材料收集容易等其他方法无可比拟的优点,并且有助于评估动物青春期的健康状况、判断动物的排卵方式、早孕诊断、产期预测等,因而是目前研究野生珍稀动物类固醇激素的一种较为理想的方法。     

拯救大熊猫赢得主动,保护大熊猫有了希望──“双控”措施是人工繁殖大熊猫的最佳选择

总结了十余年大熊猫生殖轴系垂体促性腺细胞、睾丸、卵巢、精子、卵子和受精机理以及生殖激素测定经验;分析了大熊猫生殖能力低的根本原因是生殖内分泌失调;提出了“双控”措施,即对雌性大熊猫采用补充外源促性腺激素,促进滤泡发育、卵子成熟和发情排卵,而对雄性大熊猫,采用了人工采精,精液冷冻保存的方法。采用“双控”措施的六年里圈养大熊猫空怀明显下降,获双胎7次,单胎9次,出现了大熊猫繁殖史上空前繁荣的大好形势,从此宣告拯救圈养大熊猫已赢得了主动,保护大熊猫有了希望。     

圈养雌性大熊猫发情及排卵监测方法论述及展望

国内外专家学者为监测圈养雌性大熊猫Ailuropoda melanoleuca的发情程度和排卵时间,进行了大量多学科研究,方法学的研究重点为对大熊猫的适时配种有实际指导意义。生物化学、微生物学、光谱化学和组织学等领域的研究方法,由于学科关联度低、研究深度不够、费用较高、测试时间较长等缺点,对实践指导作用有限,仅可作基础性研究。而行为学、细胞学和内分泌学等监测方法的应用更多、成效更高。大量实践也证实了通过发情行为、外阴形态、阴道细胞角化率和雌激素监测可以取得较高的受孕率,获得较好的监测效果。综合运用行为、外阴形态与雌激素监测是目前最常用的手段。本文旨在介绍圈养雌性大熊猫发情鉴定和排卵监测方法的研究进展,论述各方法的实践效果,并探讨新方法的研究方向,以期更好把握大熊猫发情配种时机。     

大熊猫感染性泌尿生殖道血尿症病原-肺炎克雷伯氏杆菌

对一只长期在秋冬季节交替或炎热夏季发生血尿症大熊猫的尿液进行细菌的分离、培养,并对分离株进行了小白鼠毒性试验和药敏试验。通过对分离菌株的染色镜检和生化试验,最后确定为肺炎克雷伯氏杆菌。作者对肺炎克雷伯氏杆菌引发大熊猫的疾病类型进行了探讨,指出肺炎克雷伯氏杆菌除可引发大熊猫肠炎和败血症外,还可导致大熊猫发生以排血样尿液为主要临床表现的泌尿生殖道感染。     

雄性大熊猫尿中睾酮和皮质醇代谢物水平及其与繁殖、笼养和季节的关系

收集了雄性大熊猫尿样肾上腺皮质激素和雄性激素代谢物年周期变化基础数据。二月份雄性激素水平最高,与繁殖季节的开始相关。繁殖成功与雄性激素水平无相关,与不同的笼养环境也无相关。与其他熊类物种相比,大熊猫交配和雄性激素水平峰值在一年中的较早时候出现。在大熊猫这样一个多配制物种,这个峰值可以看作与挑战假说相关联。在冬季,肾上腺皮质激素水平逐月升高,12月份出现峰值。在繁殖季节来临前,肾上腺皮质激素可能扮演了一个预备的角色。     

Important population viability analysis parameters for giant pandas (Aliuropoda melanoleuca)

Population viability analysis(PVA) is a tool to evaluate the risk of extinction for endangered species and aid conservation decision-making.The quality of PVA output is dependent on parameters related to population dynamics and life-history;however,it has been difficult to collect this information for the giant panda(Aliuropoda melanoleuca),a rare and endangered mammal native to China,confined to some 30 fragmented habitat patches.Since giant pandas are long-lived,mature late,have lower reproductive rates,and show little sexual dimorphism,obtaining data to perform adequate PVA has been difficult.Here,we develop a parameter sensitivity index by modeling the dynamics of six giant panda populations in the Minshan Mountains,in order to determine the parameters most influential to giant panda populations.Our data shows that the giant panda populations are most sensitive to changes in four female parameters:initial breeding age,reproductive rate,mortality rate between age 0 and 1,and mortality rate of adults.The parameter sensitivity index strongly correlated with initial population size,as smaller populations were more sensitive to changes in these four variables.This model suggests that demographic parameters of females have more influence on the results of PVA,indicating that females may play a more important role in giant panda population dynamics than males.Consequently,reintroduction of female individuals to a small giant panda population should be a high priority for conservation efforts.Our findings form a technical basis for the coming program of giant panda reintroduction,and inform which parameters are crucial to successfully and feasibly monitoring wild giant panda populations.     

Automatically detecting the wild giant panda using deep learning with context and species distribution model

The giant panda is a flagship species in ecological conservation. The infrared camera trap is an effective tool for monitoring the giant panda. Images captured by infrared camera traps must be accurately recognized before further statistical analyses can be implemented. Previous research has demonstrated that spatiotemporal and positional contextual information and the species distribution model (SDM) can improve image detection accuracy, especially for difficult-to-see images. Difficult-to-see images include those in which individual animals are only partially observed and it is challenging for the model to detect those individuals. By utilizing the attention mechanism, we developed a unique method based on deep learning that incorporates object detection, contextual information, and the SDM to achieve better detection performance in difficult-to-see images. We obtained 1169 images of the wild giant panda and divided them into a training set and a test set in a 4:1 ratio. Model assessment metrics showed that our proposed model achieved an overall performance of 98.1% in mAP_0.5 and 82.9% in recall on difficult-to-see images. Our research demonstrated that the fine-grained multimodal-fusing method applied to monitoring giant pandas in the wild can better detect the difficult-to-see panda images to enhance the wildlife monitoring system.     

Assessing vulnerability of giant pandas to climate change in the Qinling Mountains of China

Climate change might pose an additional threat to the already vulnerable giant panda (Ailuropoda melanoleuca). Effective conservation efforts require projections of vulnerability of the giant panda in facing climate change and proactive strategies to reduce emerging climate‐related threats. We used the maximum entropy model to assess the vulnerability of giant panda to climate change in the Qinling Mountains of China. The results of modeling included the following findings: (1) the area of suitable habitat for giant pandas was projected to decrease by 281 km² from climate change by the 2050s; (2) the mean elevation of suitable habitat of giant panda was predicted to shift 30 m higher due to climate change over this period; (3) the network of nature reserves protect 61.73% of current suitable habitat for the species, and 59.23% of future suitable habitat; (4) current suitable habitat mainly located in Chenggu, Taibai, and Yangxian counties (with a total area of 987 km²) was predicted to be vulnerable. Assessing the vulnerability of giant panda provided adaptive strategies for conservation programs and national park construction. We proposed adaptation strategies to ameliorate the predicted impacts of climate change on giant panda, including establishing and adjusting reserves, establishing habitat corridors, improving adaptive capacity to climate change, and strengthening monitoring of giant panda.     

岷山山系大熊猫自然保护区2003年生物多样性监测

岷山山系是全球生物多样性保护的热点地区之一,有着全世界最大的大熊猫种群和面积最大的大熊猫栖息地。到2003年底,在该地区已建立了20个以保护大熊猫为主的自然保护区。从2003年开始,岷山17个大熊猫自然保护区实施了野外大熊猫及其栖息地监测。通过监测,了解了岷山自然保护区野生动物分布状况、人为活动对自然保护区的干扰情况,以及自然保护区周边社区经济情况,为自然保护区管理决策提供了依据。同时,通过监测活动还提高了自然保护区工作人员的业务水平。建议今后将自然保护区监测工作纳入自然保护区日常工作中。     

基于高分辨率网络的大熊猫姿态估计方法

对圈养大熊猫 (Ailuropoda melanoleuca) 开展长期行为监测能及时了解其所处生理周期和健康状况,有助于繁殖饲养机构迅速采取相应繁育保护措施提高饲养管理水平,但目前无法对大熊猫进行24 h监控并及时地获得相应的行为信息。准确的动物姿态估计是动物行为研究的关键,也是诸多下游应用的基础。了解大熊猫的姿态可以促进大熊猫行为研究并提升保护管理水平。为了提高复杂环境下大熊猫姿态估计的准确率,本文以高分辨率网络 (High resolution net, HRNet) 为基础网络架构提出了一种大熊猫姿态估计方法：针对大熊猫不同部位尺度差异较大的问题,在HRNet-32中引入了空洞空间金字塔池化 (Atrous spatial pyramid pooling, ASPP) 模块,在提升特征感受野的同时捕获多尺度信息;同时对大熊猫身体关键点进行分组,引入基于部位的多分支结构来学习特定于每个部位组的表征。多次对比实验结果表明本文所用模型具有较高的检测精度：在PCK@0.05中所用模型精度达到了81.51%。本文提出的方法可为大熊猫的行为分析和健康评估提供技术支撑。     

从长期监测数据看佛坪自然保护区大熊猫种群的发展趋势与生态习性

野生动物监测在保护管理自然资源以确保可持续利用上占有关键地位,在西方一些发达国家已成为濒危物种乃至生物多样性保护的一项重要内容。在我国,对大型哺乳动物的长期监测基本尚未纳入常规,针对野生种群的监测报道迄今罕见。本文以佛坪自然保护区2000 ～ 2006 年大熊猫监测数据为基础,通过大熊猫遗留的新鲜粪便等痕迹指标探讨了该地大熊猫种群的发展趋势与监测数据所反映的大熊猫生态习性。结果显示该保护区内大熊猫种群略显缓慢增加的趋势(t = 1.98,P = 0. 082),不同季节痕迹的海拔变化表明,该地大熊猫有季节性垂直迁移的生态习性。此外,监测数据所反映的大熊猫空间分布等亦与其他相关研究大致吻合,表明该保护区所采用的监测方法有在全国其他大熊猫保护区推广利用的潜在价值。     

大熊猫等五种食肉动物血清蛋白和LDH同工酶盘电泳比较

大熊猫(Ailuropoda melanoleuca)是世界珍稀动物之一,近年来,兽类科学工作者对它进行了多方面的研究,也包括生化方面的工作(潘文石等,1982;Sarich,1973),但与其他食肉哺乳动物相比研究较少,而此又为探讨其分类地位所需。本文应用聚丙烯酰胺凝胶盘电泳,对大熊猫、小熊猫(AilurSs fulgens)、黑熊(Selenarctos thibetanus)、家猫及犬等5种食肉动物的血清蛋白和LDH同工酶进行比较分析,目的在于了解这5种动物的血清蛋白图象和LDH同工酶酶谱以及它们之间的谱型、相对活力、迁移率的异同,进而讨论大熊猫的分类地位。     

Papanicolaou staining of exfoliated vaginal epithelial cells facilitates the prediction of ovulation in the giant panda

The giant panda is seasonally monoestrus, experiencing a single estrous with spontaneous ovulation in the spring. Therefore, accurate monitoring of the estrous cycle to pinpoint the time of ovulation is critical for the success of timed mating or artificial insemination. Analysis of exfoliated vaginal epithelial cells is a simple technique that rapidly yields information about the estrous status of a panda. Vaginal swabs were obtained during five estrous cycles of two nulliparous females. Cells were stained with the trichrome Papanicolaou and classified as basophils, intermediates or superficials. The color of stained cells, basophilic, acidophilic or keratinized, was recorded as a characteristic independent of the three standard cell types. The day urinary conjugates of estrogen fell from peak levels was considered the day of ovulation. A chromic shift occurred 8-9 days before ovulation when the majority of exfoliated vaginal cells changed from basophilic (blue) to acidophilic (pink) without accompanying nuclear or cytoplasmic changes. A second chromic shift was consistently observed 2 days prior to ovulation when keratinized (orange) cells replaced acidophils as the majority of vaginal cells. Monochrome staining of vaginal cells is sufficient to quantify superficial cells, which is a useful adjunct to behavioral and endocrinological data in determining estrous in the giant panda. However, the timing and duration of superficial cell elevations are substantially different between and within individual females, which limits the accuracy of timing ovulation for artificial insemination. The predictive value of vaginal cytology was greatly enhanced with the trichrome stain and evaluation of cell color.