幼年长江江豚的行为谱(英文)

采用焦点动物跟踪法及连续记录法,观察了人工饲养条件下一头幼年长江江豚(Neophocaena phocaenoides asiaeorientalis)的水下以及半自然条件下两头幼豚的水面行为型式。观察时间最长持续至出生后一年,以构建其行为谱。结果显示:1)所有幼豚都能表现出丰富多样且活跃的行为型式;2)幼豚刚出生时,与存活攸关的行为型式最先出现;3)具有玩耍和社居性质的行为型式在行为谱中占大部分;4)幼豚大部分行为型式已与其他年龄段个体的相似;5)大部分行为型式出现在生命的早期阶段。以上特征可能是这种动物对水生和社居生活的适应。     

Development and characterization of polymorphic microsatellite loci in the endangered Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis)

The finless porpoise (Neophocaena phocaenoides) is one of the smallest cetacean species widely distributed in the shallow coastal waters of the Indo-Pacific Oceans. The population size of the Yangtze subspecies (N. p. asiaeorientalis) has sharply decreased in the last two decades and access to objective data on its population structure and genetic diversity would be of great assistance for their proper management. Here we report on the isolation of nine polymorphic microsatellite using the “Fast Isolation by AFLP of Sequences Containing repeats” (FIASCO) protocol. Polymorphism was assessed using 30 porpoise individuals randomly sampled in the Yangtze River. The number of alleles per locus varies from 2 to 9, with an average value of 5.56, whereas the ranges of observed and expected heterozygosities were 0.300–0.633 (mean 0.496) and 0.473–0.804 (mean 0.659), respectively.     

基于PAE编码系统构建的半自然条件下长江江豚行为谱

识别和编制动物行为谱是深入研究动物行为及其与环境复杂关系的基础和前提。本研究于2013年8月至2014年4月以栖息在铜陵淡水豚国家级自然保护区半自然水域的8头长江江豚为研究对象,采用焦点动物取样法和随机动物取样法观察记录了动物行为发生的过程、内容和环境,并以"姿势—动作—环境"(posture-act-environment,PAE)为轴心,以行为的功能为依据,构建了长江江豚的PAE行为谱。共分辨记录了半自然水域长江江豚的8种姿势、33种动作和46种行为,并定性描述了不同行为出现频率与年龄组、性别的关系。该行为谱综合了姿势和环境信息,有助于进一步研究长江江豚的生态与行为、促进长江江豚的保护。     

Respiratory pattern of captive Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis)

Cetacean respiration usually happen in bouts. The most widely applied quantitative method used to analyze the structure of these bouts is the log_e-survivorship analysis, based on the assumption that the respiratory intervals are distributed as negative exponentials. However, for the data collected from three captive Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis), we failed to obtain a convergent result with the application of log_e-survivorship analysis. However, the two-Gaussian model, which was recently proposed to analyze the feeding behavior of cows, was successfully fitted to the data. According to the fitting results, the overall respiratory pattern of the captive Yangtze finless porpoises can be described as a dive with a mean duration of around 30–40 s, followed by two or three ventilations with a mean interval of approximately 9 s. The average intra-bout intervals during both active and inactive periods are constant at 7.7–9.9 s for all individuals. However, when shifting from active to inactive states, the adult male and female decrease their mean numbers of respirations per bout and average length of inter-bout respiratory intervals, while the estimates of both parameters increase for the juvenile female. It was pointed out that the two-Gaussian model might be more adequate for cetacean respiratory-bout structure analyses than the log_e-survivorship technique.     

半自然条件下成年雌性长江江豚的抚幼行为

2014年5-9月,采用目标动物取样法,对栖息在铜陵淡水豚国家级自然保护区半自然水域的两头成年雌性长江江豚分娩后4个月内的抚幼行为进行观察。结果表明,母豚有5种明显的抚幼行为,其中以护领行为和携带行为为主,托举行为、返回行为和干涉行为发生频率较低;随着幼豚发育,托举、携带、干涉行为呈下降趋势;护领行为先上升,后趋于平稳;返回行为变化相对平稳,但在幼豚2月龄左右出现一个高峰。此外,两头母豚间的干涉和返回行为发生频率差别较大。综合以上结果可见,出生后约两个月内是半自然水域长江江豚抚幼的关键时期,应在此期间加强管理。     

Analysis of the bacterial diversity in the fecal material of the endangered Yangtze finless porpoise, <Emphasis Type="Italic">Neophocaena phocaenoides asiaeorientalis</Emphasis>

The aim of this study was to determine the bacteria present in the fecal material of the endangered Yangtze finless porpoise, Neophocaena phocaenoides asiaeorientalis. Fecal samples were collected from 12 Yangtze finless porpoises living in the wild at Poyang Lake, located in Jiangxi Province, China. To determine the bacterial diversity, a 16S rRNA gene clone library using the bacterial PCR primers fD1 and rP2, was prepared. A total of 138 near-full-length sequences were analyzed and 39 operational taxonomic units (OTUs) were identified. Sequences showing ≥97% similarity were grouped together as an OTU. Six different phyla were identified in which 38 OTUs were classified. Most of the OTUs contained sequences belonged to the phylum Firmicutes (51.3%), followed by Tenericutes (17.9%), Proteobacteria (15.4%), Actinobacteria (7.7%), Deinococcus-Thermus (2.6%) and Cyanobacteria (2.6%). A phylum could not be assigned for one clone within one OTU (2.6%). It appears that the Yangtze finless porpoise has a more diverse range of bacteria compared to other aquatic mammals, such as seals.     

Physiological responses to capture and handling of free-ranging male Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis)

We captured free-ranging male Yangtze finless porpoises over three seasons and assayed leukocytes and serum biochemistry to investigate physiological responses to the capture and handlings. Serum thyroid hormones (THs) declined sharply in those porpoises compared with hormone variation in a captive male finless porpoise. Hypernatremia and hypokalemia were also significant in the free-ranging animals suggesting that conservation of serum sodium might be acutely vital for this freshwater subspecies. The animals captured in spring showed more significant neutrophilia and eosinopenia than those captured in autumn suggesting that they may be more affected by capture during the breeding season. Furthermore, physical examination of porpoises when out of the water was apparently stressful, particularly when they were kept out of the water for longer periods. However, an increase in circulating THs may be an adaptive response to accommodate these short-term stresses.     

Isolation and characterization of microsatellite loci in the finless porpoise (Neophocaena phocaenoides)

Eight dinucleotide microsatellite loci were isolated from the finless porpoise (Neophocaena phocaenoides). Analysis of 30 individuals showed the number of alleles ranged from four to 21 with observed heterozygosity ranging from 0.300 to 0.833, and expected heterozygosity ranging from 0.437 to 0.932. Cross‐species amplification was tested in four other cetacean species. These microsatellite markers would be valuable tools for population genetic studies of finless porpoises and other cetacean species.     

长江江豚和瓶鼻海豚的血清转铁蛋白

血清转铁蛋白(Serum transferrin,Tf)是广泛存在于血清中的β球蛋白,其主要功能是将铁离子以安全的形式(Fe3 )在机体的吸收、储存和利用部位之间进行传递,为细胞生长提供铁(Fe3 )(Welch,1990);同时具有保护细胞、促进细胞分化和生长、抑菌和杀菌的作用(Paezetal.,2002;Liuet al     

Herpes-like virus infection in Yangtze finless porpoise (Neophocaena phocaenoides): pathology, ultrastructure and molecular analysis

A moribund juvenile Yangtze finless porpoise (Neophocaena phocaenoides) with skin lesions and ulceration was found in Dongting Lake, China. Pathologic examination, electron microscopy, and polymerase chain reaction of liver tissue revealed widely distributed necrotic lesions, sinusoidal dilatation, congestion and herpes-like virus particles.     

用微卫星指纹识别天鹅洲保护区长江江豚个体

DNA指纹个体识别技术是保护遗传学研究中的一种非常重要的手段。为了准确地识别天鹅洲保护区中的每一头长江江豚以开展保护遗传学及其它相关研究 ,并实施有效的种群管理 ,本研究应用 4个微卫星座位初步构建了该群体的DNA指纹图谱 ,并利用此图谱成功地对不同时期在保护区捕获的江豚进行了个体识别研究。结果显示微卫星指纹技术是一种适用于长江江豚个体识别研究的可靠手段     

辽宁海域东亚江豚的年龄鉴定与生长规律

对2007-2018年辽宁海域发现的39头死亡东亚江豚(Neophocaena asiaeorientalis sunameri)进行了外部形态参数测量和生长规律分析,并通过观察牙齿中的生长层组对2014-2018年的27头江豚个体进行了年龄鉴定.结果表明:死亡江豚以0~4龄个体为主,雄性21头,雌性18头;同龄雌雄个体间的体长、体重均无显著差异(P>0.05);所有江豚样本的体重与年龄(Y=137e^-2.475exp(-0.205t))、体长与年龄(Y=232(1-0.680e^-0.161t)均极显著相关(P<0.01),且随年龄增大,体重、体长均增加但增速下降.死亡江豚的发现时间在不同的海域存在差异:黄海北部海域集中在每年的3-6月,以大连星海湾至老虎滩沿岸为多;渤海的辽东湾海域主要在每年的10-11月初,多发现在大连长兴岛以北的海域.辽东湾海域江豚的死亡多发生在渔业作业期,可能与渔业生产活动有关.研究还发现,在一年中辽东湾海域死亡江豚集中出现的时间与斑海豹的分布期存在交替现象,是否与两种海洋哺乳动物适应海水温度能力、种群间的食物竞争以及季节性迁移等因素有关还有待于进一步的研究.     

Genetic variation of the MHC DQB locus in the finless porpoise (Neophocaena phocaenoides)

The Major Histocompatibility Complex (MHC) is a large multigene coding for glycoproteins that play a key role in the initiation of immune responses in vertebrates. The exon 2 region of the MHC DQB locus was analyzed using 160 finless porpoises from 5 populations in Japanese waters. The 5 populations were based on a previous mitochondrial DNA control region analysis, which showed distinct geographical separation. Eight DQB alleles were detected, and the geographical distribution of the alleles indicated that most of them are shared among the populations. Heterozygosity of the DQB alleles in each population ranged from 0.55 to 0.78, and for all 5 populations was 0.78. Low MHC variability is not a common feature in marine mammals, but the finless porpoise populations inhabiting coastal waters had a relatively high MHC heterozygosity. Balancing selection in the MHC DQB alleles of the finless porpoise was indicated by the higher rate of nonsynonymous than synonymous substitutions for PBR; however, an excess of hetrozygotes compared to expectation was not observed. This suggests that the MHC DQB locus in the finless porpoise may have been under balancing selection for a long evolutionary time period, and is influenced by genetic drift beyond the effect of balancing selection for short time periods in small local populations.     

SNP discovery based on CATS and genotyping in the finless porpoise (Neophocaena phocaenoides)

Single nucleotide polymorphisms (SNPs) discovery and genotyping were performed for the finless porpoises (Neophocaena phocaenoides). About 202 comparative anchor tagged sequence primers derived from genomes of human, mouse and some other mammals were used to screen the finless porpoise population. Of the 51 SNPs discovered, 25 were further characterized with ideal genotyping primers and using fragment length discrepant allele specific PCR assay. This is the first report of SNP loci for the finless porpoise, which is helpful to provide some novel molecular markers and new genetic information relevant to the conservation and management of this endangered species.     

基于CATS法的江豚SNPs位点筛选

通过比较锚定序列示踪(Comparative Anchor Tagged Sequences, CATS)法,选择人、小鼠及其它哺乳动物中已知的5对CATS引物,通过PCR反应从江豚(Neophocaena phocaenoides)基因组中扩增出相应的DNA片段,结合变性高效液相色谱(Denaturing High Performance Liquid Chromatography, DHPLC)和DNA直接测序等技术,进行江豚单核苷酸多态性(Single nucleotide polymorphisms, SNPs)位点的筛选.通过不同个体同源序列的比对分析,发现其中的4个DNA片段具有多态性,共检测出7个SNPs位点,即大约每370 bp出现1个SNPs.本研究为进一步利用CATS法获得江豚及其它野生动物种群的SNPs位点并进行种群遗传学分析奠定了基础     

微卫星分型法应用于豢养繁殖长江江豚的父权鉴定

目的:鉴定武汉白鱀豚馆及安徽铜陵淡水豚保护区两个豢养长江江豚繁殖群体中出生的6头幼豚的生物学父亲.方法:选择8对江豚物种特异性微卫星引物对两个待鉴定群体的14个DNA样品进行了荧光标记PCR扩增,将纯化后的扩增产物在ABI3130遗传分析仪上进行基因分型,并根据GeneScan Rox 500内标确定不同等位基因的大小,随后对待鉴定对象进行等位基因分析,并计算主要多态性参数.结果:所采用的8个微卫星座位在待鉴定的两个江豚群体中均表现出不同程度的多态性.在母本已知的条件下,利用其中6个微卫星座位的等位基因数据,通过排除法成功地鉴定出两个繁殖群体中出生的6头幼豚的生物学父亲.结论:本研究首次成功地将6个物种特异性微卫星标记应用于豢养长江江豚的父权鉴定,从而为该物种微卫星亲子鉴定技术体系的建立及迁地保护繁殖群体遗传谱系的构建奠定了技术基础.     

豢养长江江豚性行为的观察

2001年12月至2002年11月作者采用焦点动物取样法对豢养池中3头长江江豚(1♂,2♀)的性行为进行了持续1年的水下观察。结果表明：雄性江豚生殖器全年均有伸出行为,4～5月和9-10月为高峰期。其生殖器伸出方式主要有侧伸、弓伸、仰伸、下伸和竖伸5种方式。豢养池中的气压、光照、池水透明度和pH值能基本满足江豚性活动的要求。     

High MHC DQB Variation and Asymmetric Allelic Distribution in the Endangered Yangtze Finless Porpoise, Neophocaena phocaenoides asiaeorientalis

The endangered Yangtze finless porpoise is found in the middle and lower reaches of the Yangtze River and its adjoining big lakes. To explore the major histocompatibility complex (MHC) genetic diversity and allelic distribution patterns across its range, we investigated variation at DQB exon 2. From 76 porpoises, we identified 18 DQB sequences. The freshwater Yangtze populations had much higher allelic diversity than marine populations. Among these freshwater populations, the middle-reach population had higher allelic diversity than the lower-reach population. The high DQB diversity level, relative to that of a neutral mtDNA locus, suggests that balancing selection is acting at the DQB gene and that rapid evolution and local positive selection play critical roles in generating and retaining high MHC diversity in the freshwater population. As the balancing selection might be driven by environmental pathogens, we suggest that maintaining MHC variation should be a high priority in the conservation and management of this endangered population, especially as an ex situ conservation strategy.     

Population Genetic Structure of the Yangtze Finless Porpoise (Neophocaena phocaenoides asiaeorientalis): Implications for Management and Conservation

Understanding the population genetic structure is a prerequisite for conservation of a species. The degree of genetic variability characteristic of the mitochondrial DNA control region has been widely exploited in studies of population genetic structure and can be useful in identifying meaningful population subdivisions. To estimate the genetic profile of the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis), an endangered freshwater population endemic to China, the complete mtDNA control region was examined in 39 individuals belonging to seven different stocks inhabiting the middle and lower reaches of the Yangtze River. Very low genetic diversity was found (nucleotide diversity 0.0011± 0.0002 and haplotypic diversity 0.65± 0.05). The mtDNA genetic pattern of the Yangtze population appears to indicate a founder event in its evolutionary history and to support the marine origin for this population. Analyses by F_st and Φ_st yielded statistically significant population genetic structure (F_st = 0.44, P < 0.05; Φ_st = 0.36, P < 0.05). These results may have significant implications for the management and conservation of the Yangtze finless porpoise in the future.