Molecular polymorphism of MHC-DRB gene and genetic diversity analysis of captive forest musk deer (Moschus berezovskii)

The major histocompatibility complex (MHC) is one of the most important elements in immune system for nearly all vertebrates, and is thought to be essential for an organism to recognize foreign molecules. In this study, we investigated the MHC variation in 51 forest musk deer (Moschus berezovskii) collected from three captive populations in Sichuan Province, China. Seventeen haplotypes were isolated from the 51 samples. A total of 51 mutation sites were identified and yielded a nucleotide diversity of 0.056. These haplotype sequences possessed 83 putative amino acid sites, including 24 PBR sites (peptide binding region) and 59 non-PBR sites. Out of 24 PBR sites, 15 codons showed variation (0.625), while 12 codons showed variation (0.203) in 59 non-PBR sites. Non-synonymous substitutions primarily occurred in PBR, with analyses suggesting that the Mobe-DRB gene had undergone strong positive selection during their evolution. Compared with that of some other endangered species, the forest musk deer had relatively high level of MHC diversity. Our results suggested that the MHC diversity characteristic of captive forest musk deer populations might be not responsible for their high morbidity of abscess disease.     

凉山山系林麝夏季利用生境特征

采用样线法对位于四川凉山山系的美姑大风顶自然保护区林麝(Moschus ber-ezovskii)的夏季利用生境特征进行了调查,并对利用生境样地(n=44)和非利用样地(n=209)的生境变量进行了比较.结果表明:大风顶保护区的林麝夏季利用生境的郁闭度(32.4%±1.9%)、灌木盖度(27.1%±2.4%)及竹子盖度...     

Genetic diversity of captive forest musk deer (Moschus berezovskii) inferred from the mitochondrial DNA control region

Forest musk deer ( Moschus berezovskii ) were once distributed widely in China. However, wild populations have declined dramatically because of poaching and habitat loss. Captive breeding populations have been established for several decades, but the genetic backgrounds of most captive populations were unclear and the population sizes increased very slowly. To provide useful information for conservation and management of this species, we investigated the genetic diversity and population structure of forest musk deer by analysing a 582-bp fragment of the mitochondrial DNA (mtDNA) control region (CR) in three captive breeding populations in Sichuan Province, China. Ninety-four variable sites and 27 haplotypes were observed in 109 individuals, and the nucleotide and haplotype diversities were relatively high compared with those of other endangered mammals. Of the three investigated populations, the Maerkang population had the highest nucleotide diversity ( π  = 0.0568), haplotype diversity ( h  =   0.836) and average intra-population genetic distance (0.062). The analysis of molecular variance demonstrated that most variation occurred within samples and that there was significant differentiation of the three populations. Estimates of gene flow indicated that there were few genetic exchanges among the three populations. Building pedigree records and increasing gene flow between populations will be helpful for conserving these populations and this species.     

圈养林麝社会等级与粪样类固醇激素水平的关系

动物社群的社会等级可对个体的资源分配、社群稳定及个体的行为对策产生重要影响,圈养动物的类固醇激素水平与饲养管理有关,可反映动物的社群紧张水平.深入理解圈养动物的社会等级与其类固醇激素水平的关系是进行濒危动物迁地保育和成功驯养的基础.2018年6月15日—8月15日,用焦点取样法对四川马尔康林麝繁育场的25头林麝进行了冲...     

Isolation and characterization of microsatellite loci from forest musk deer (Moschus berezovskii)

This study reported the isolation and characterization of eight polymorphic microsatellite loci in endangered forest musk deer Moschus berezovskii. An improved enrichment protocol was used to isolate microsatellites, and polymorphism was explored with samples from wild musk deer population collected in Miyalo of Sichuan Province in China. Approximately 70% of clones from the genomic library constructed in current study contained dinucleotide (AC) repeats. Eight microsatellite loci amplified were highly polymorphic within forest musk deer population. The number of alleles per locus ranged from 6 to 14, and the observed and expected heterozygosities ranged from 0.41 approximately 1.0 and from 0.8 approximately 0.9, respectively. The average polymorphic information content (PIC) value for these markers was 0.82. This demonstrated that the eight microsatellite loci developed here are highly polymorphic, and can be used as genetic markers for further investigation of musk deer. Also, the results showed that the musk deer distributed in Miyalo had a relatively higher level of genetic variation.     

原麝和林麝冬夏两季粪便真菌菌群多样性

肠道微生物群对宿主健康的影响取决于饮食环境和宿主等因素.本研究通过Illumina MiSeq平台对冬、夏两季林麝和原麝粪便真菌ITS1区段进行测序,探究原麝和林麝肠道真菌菌群结构特性及季节因素对其多样性的影响.结果 表明,林麝和原麝肠道真菌组成中子囊菌门Ascomycota均为优势菌门,它们的相对丰度随物种和季节因素...     

圈养林麝的活跃性及与社会亲和度的关系

采用扫描取样和焦点动物取样法对四川马尔康林麝繁育场的45头圈养林麝进行行为取样,以其非静息行为的发生率作为活跃性指标,以社会亲和行为的发生强度作为社会亲和度指标,分析圈养林麝活跃性与其性别、圈群密度及社会亲和度的关系。结果表明:圈群密度和社会亲和度对圈养林麝的活跃性有显著影响(P < 0.05),但活跃性雌雄差异不显著(P > 0.05)。与高密度圈群的林麝(0.25 ± 0.021, n=32)相比,低密度圈群中的个体更活跃(0.40 ± 0.022, n = 4);活跃性与社会亲和度之间显著相关(r = 0.390, P = 0.01),活跃性较高个体倾向于展现高的社会亲和度。建议麝场在进行麝群构建时,充分考虑林麝个体的活跃性差异及与圈群密度的关系,以提高圈群稳定性和驯养生产力。     

The reproductive performance of female Forest musk deer (Moschus berezovskii) in captivity

Reproductive performance of 750 adult female Forest musk deer (FMD, Moschus berezovskii) was monitored (from 2005 to 2009) on two Forest musk deer farms in Sichuan province, China. The mean (± SEM) lengths of the estrous cycle, pregnancy, and consecutive fawning intervals were 17.5 ± 0.8 d (n = 64), 182.9 ± 0.8 d (n =190), and 363.2 ± 2.4 d (n = 120), respectively. Copulation occurred from the end of October to early March, with the majority (84.1 ± 1.9%) during November and December (P < 0.05). The fawning rate was 68.7 ± 2.2% in yearling hinds and 73.3 ± 1.3% in adult hinds. Fawning occurred from the end of April to early September, with the majority (88. 9 ± 2.2%) during May and June (P < 0.005), including a peak in May (52.0 ± 1.0%). Yearling hinds fawned later (7.3 ± 1.8 d) than adult hinds. Rates of premature delivery and dystocia were 2.7 ± 0.4 and 2.0 ± 0.3%, respectively. There was a 1.07:1 female-to-male ratio at birth (P > 0.05), with 67.7% twins, but only two cases of triplets and one set of quadruplets among 537 fawns. Birth weight averaged 518.7 ± 10.3 g, ranging from 304 to 775 g (n = 240), with male fawns slightly heavier than females (521.1 ± 14.8 vs 516.9 ± 14.3 g). Postnatal and pre-weaning mortality rates were 25.0 ± 1.2 and 17.9 ± 3.1%, respectively. Postnatal mortality and urinary calculus were much more common in male fawns (P < 0.01). The mortality rate due to suppuration and dyspepsia were different (P < 0.001) between postnatal and pre-weaning periods. Due to higher mortality of male fawns, the adult sex ratio was significantly female-biased. In conclusion, captive FMD had similar seasonal patterns of copulation and fawning as wild FMD and other musk deer species.     

圈养林麝春季活动节律及时间分配

2006 年春季,使用红外夜间观察记录仪,对崇明东平麝场12 只成年林麝(8 雌,4 雄)的行为进行了24 h昼夜连续观察。发现春季林麝的行为有37 种,可归并为休息、摄食、运动、社会、维持和排遗6 大行为类型,据此,建构了春季林麝的行为谱。根据各行为持续时间比率和频次分析表明,林麝的主要行为为休息;雌麝的摄食行为所占的比率高于雄麝,这是由于春季雌麝妊娠的能量需求较高所致。活动节律呈典型的晨昏双峰型,清晨(4:00 ～ 6:00)和傍晚(17:00 ～ 19:00)为活动高峰;夜间活动性显著高于白天。     

Genetic quality of the Miyaluo captive forest musk deer (Moschus berezovskii) population as assessed by microsatellite loci

The Miyaluo captive forest musk deer population (Sichuan Province, China) is one of the largest captive breeding populations in the world. In order to evaluate the genetic quality and provide available genetic management strategy, seven polymorphism microsatellite loci were applied to assess the genetic variation of the Miyaluo forest musk deer. The results indicated that a total of 168 alleles were detected from these seven microsatellite loci in 361 individuals, and the number of the alleles per locus ranged from 12 to 41 with a mean of 24. The average observed heterozygosity, expected heterozygosity, and PIC were 0.782, 0.854, and 0.837, respectively. Considering the results of the loci Hardy–Weinberg equilibrium test, the comparison of the common allele frequency as well as the private allele between the adults and juveniles, we concluded that the heterozygosity and the genetic diversity of the Miyaluo captive breeding population are increasing due to the input of new individuals from other populations. However, the frequency of some alleles declined sharply, and some were even lost indicating that there is a risk for diversity loss. Thus, we proposed an improved management and breeding strategy for the captive breeding population of the forest musk deer.     

林麝及其近缘物种编码区微卫星分布规律及功能分析

麝科和鹿科动物均属于偶蹄反刍类动物,具有重要的经济价值。通过系统的微卫星序列 (Simple sequences repeats, SSRs) 从基因组水平揭示物种间的系统进化关系,探索微卫星序列的基因功能及其富集的信号通路,目前仍缺乏相关研究。随着林麝 (Moschus berezovskii)、原麝 (Moschus moschiferus)、小麂 (Muntiacus reevesi)、赤麂 (Muntiacus vaginalis) 和马鹿 (Cervus elaphus) 基因组测序的完成,本文利用生物信息学方法提取了这些动物蛋白质编码区 (coding sequences, CDS) 序列,统计和分析了其CDS区微卫星序列分布规律及其生物学功能,探索了含SSR 基因富集的信号通路及其与疾病的关联性。结果表明,林麝、原麝、小麂、赤麂和马鹿蛋白质编码区含SSR序列的基因所占比例分别为6.96% (1 696个)、7.18% (2 359个)、7.29% (3 005个)、7.36% (1 916个) 和7.48% (1 924个),并且这5种动物CDS区SSRs分布模式具有相似性,均是三倍体核苷酸 (即三核苷酸和六核苷酸) SSRs最多,分别为96.85%、94.87%、65.44%、64.23%和88.04%。GO功能富集表明,林麝与其他4种动物蛋白质编码区SSR序列在分子功能、细胞组成和生物学过程3个方面具有较多共同显著富集的功能,包括DNA结合、染色质和生长发育等。KEGG 通路分析表明,林麝及其他4种动物蛋白质编码区SSR序列具有7个共同显著富集的KEGG通路,包括遗传信息调控蛋白家族、转录因子、染色体及相关蛋白、剪接体、转录机制和Notch信号通路和成体糖尿病。通过对林麝编码区含SSR关键免疫基因及其相关联的KEGG通路进行分析,发现10个含SSR的关键免疫基因对应的KEGG通路与疾病密切相关。     

圈养林麝粪样类固醇激素水平与麝香分泌的关系

基于非损伤取样和放射免疫法(RIA),于2018年9月1日—10月15日期间对四川马尔康林麝繁育场的36头雄性圈养林麝(Moschus berezovskii)的粪样类固醇激素水平进行了检测,结合麝香分泌数据采集,探究了林麝的粪样睾酮和皮质醇水平与其麝香分泌的关系,结果表明:四川马尔康圈养林麝的麝香分泌(12.53±0.76) g (n=36)与粪样睾酮水平(106.03±12.34) ng/g (n=36)呈显著正相关(r=0.436,P<0.01),泌香较多雄麝(16.50±0.61) g (n=17)的睾酮水平(141.51±21.86) ng/g (n=17)显著地高于泌香较少(8.98±0.59) g (n=19)雄麝(74.27±7.93) ng/g (n=19)(t=-3.252,df=34,P<0.01),方程y=0.021x+10.270(R²=0.120,P<0.05)可较好地拟合二者间关系。麝香分泌与皮质醇浓度(47.49±1.53) ng/g (n=36)相关不显著(r=0.078,P>0.05),泌香较多雄麝的皮质...     

The olfactory subgenome and specific odor recognition in forest musk deer

Olfactory receptors (ORs) are encoded by OR genes. The OR genes in forest musk deer (Moschus berezovskii), which rely on olfaction for reproductive and social communication, are poorly understood. In this study, we analyzed the genome sequence of the forest musk deer to obtain its olfactory subgenome and compared it to other species. A total of 1378 OR‐related sequences were detected in the forest musk deer genome including 864 functional genes, 366 pseudogenes and 148 partial genes. These OR genes were classified into Class I and Class II and were further classified into 18 families and 244 subfamilies through sequence identity. Comparative analyses of the OR genes’ protein sequences in species from different orders (forest musk deer, human, mouse and dog) showed that 12 clusters were specific to forest musk deer. However, when compared to other Artiodactyl species (i.e. cattle, yak and pig) only two clusters were specific to forest musk deer. The odor identification potential of the OR genes in the forest musk deer was focused mainly on floral, woody, lemon, sweet and fatty odors. We also found that OR genes specific to forest musk deer were involved in the identification of spearmint and caraway. Our work is the first genome‐wide analysis of OR genes in forest musk deer. These findings will assist with better understanding the relationship between behavior and olfaction in the forest musk deer and the characteristics of the olfactory subgenome in Artiodactyl mammals.     

林麝幼体的换毛序和胎毛、夏毛、冬毛的形态

被毛是哺乳动物最主要的特征之一,有关动物换毛序与环境关系的研究,是动物生态学研究的一个重要方面。从1853年Browne开始研究毛发结构以来,人们对毛发的形态学、分类学和种属关系等方面的研究已开展了大量工作,特别是随着电镜技术的应用,这些方面的工作在逐渐深入(曹汉民等,19     

林麝(Moschus berezovskii)研究概况和进展

林麝（Moschus berezovskii）是国家一级保护动物,已被列入CITES附录Ⅰ中,由于连续多年的恶性捕杀,加上国际高价走私,使中国的野生林麝资源遭到严重破坏。作者根据大量文献资料结合自己的研究,从林麝的分类地位、地理分布、组织解剖、种群生态和人工饲养等几个方面较为全面地总结了国内林麝研究的概况和进展,指出了林麝种群研究中存在的一些问题以及今后的研究方向。     

小兴安岭通河林区原麝夏季对生境的选择

2004年8 ～ 9月,在通河县龙口林场采用样线样方法对原麝夏季生境的选择进行了研究。共设235个样方,其中原麝利用样方55 个,采用卡方（Chi-square） 统计进行显著性检验,结果表明原麝偏好利用针阔混交林,避免杂木林;倾向于选择离水源近、远离人为干扰的上坡位;一般喜欢有石砬子的山坡;对坡向的利用无选择性。对原麝利用样方和非利用样方进行比较,发现原麝生境具有海拔高、郁闭度小、食物丰富和坡度陡等特征。对各种生境因子的主成分分析表明,前6个特征值的累积贡献率达到73.30%,可以较好地反映原麝生境特征,根据载荷系数绝对值大小将原麝夏季生境选择影响因子分别命名为食物丰富度因子、植被因子、干扰因子、隐蔽度因子、地理因子和倒木因子。     

Development and characterization of 15 novel microsatellite markers from forest musk deer (<Emphasis Type="Italic">Moschus berezovskii</Emphasis>)

A total of 15 polymorphic microsatellite loci were isolated from forest musk deer (Moschus berezovskii). These loci showed high levels of allelic diversity, presenting 6–14 alleles per locus in a group with 31 individuals. The mean observed and expected heterozygosities were 0.82 and 0.68, respectively. The average polymorphic information content value, cumulative discrimination power, overall probability of exclusion with both unknown parents or with only unknown sire for these markers were 0.78, 1.0, 0.9999 and 1.0, respectively, indicating that this suit of microsatellite loci would be a powerful tool for the genetic determination of the forest musk deer.     

甘肃兴隆山林区马麝(Moschus chrysogaster)隔离种群的生态特征

兴隆山林区大约在300a前就已完全隔离。现有马麝种群的密度高而相对稳定,约为30－50头／km^2,在目前密度与系统压力下,种群数量基本稳定在5000头左右,但却面临着疾病、营养不良与偷猎的威胁。因此应努力建立一个“以开发养保护,以保护促发展”的良性循环模式,从而为我国自然保护区的建设和发展树立典范。     

圈养林麝麝香分泌时间节律及影响因素

在2016年和2017年的5—8月,我们对川西马尔康麝场圈养林麝（Moschus berezovskii）的麝香分泌进行了行为与生理监测,对麝香分泌的各阶段进行了准确判定,记录了泌香启动、泌香盛期开始、泌香盛期终止及泌香结束的时间阶段及持续时间长,分析了林麝麝香分泌的时间阶段与体重和年龄等因素的关系。结果表明,马尔康麝场的雄性林麝平均泌香启动日在6月16日[（167.06±7.75）d,n=141],于6月17日进入泌香盛期[（168.52±7.67）d,n=141],6月21日[（172.17±7.26）d, n=138]泌香活动减弱,至6月25日[（176.27±8.11）d, n=131]泌香结束;雄麝体重与其泌香启动、泌香盛期停止及泌香结束时间呈显著负相关（r_PS = -0.234,P_PS =0.028;r _VSF = -0.215,P_VSF = 0.047;r _SE = -0.229,P_SE = 0.043）,即雄麝的体重越大,其泌香越早;各年龄组间的平均泌香时长差异显著（F _{17, 113} = 3.482, P = 0.003）,其中2岁雄麝平均泌香时长最长[（13.07±2.08）d, n=20],显著高于3岁[（9.38±0.76）d, n=12, P = 0.042]和4岁[（7.80±1.60）d, n=5, P = 0.013]个体;马尔康林麝平均泌香量为（11.85g±0.96）g, （n=114）,随泌香时长延长有增加趋势,但不显著（P = 0.854）。基于上述林麝雄体的泌香时间、泌香量与年龄和体重等因素间的关联,可对圈养的林麝个体间的泌香力、泌香量等进行区分和预判,作为圈养林麝驯养生产力优化的依据,并可为圈养林麝优质品系的选育提供参考。     

兴隆山自然保护区马麝春季生境选择

于2006年3月和2007年4月,采用样线样带结合调查法,在甘肃兴隆山自然保护区对野生马麝(Moschus Sifanicus)的春季生境选择和栖息地利用格局进行了研究,共布设280个空白对照样地和52个利用样地,对海拔和郁闭度等17个生境变量进行了计测和比较,结果表明,与对照的非利用样地相比,兴隆山马麝春季利用样地的海拔相对较低(2280.94±17.06)m、乔木平均高度较高(9.36±1.21)m、乔木胸径较大(102.44±14.25)cm及地表植被盖度较好(26.67±3.27)%,上述差异均达显著程度(P0.05);此外,马麝春季趋于利用南坡(44.23%)和西坡(36.54%)的中下坡位(86.54%)的坡度适中(30°—60°,55.77%)的生境,其避风性(63.46%)和隐蔽度(76.92%)均较差,距离水源(1000m,80.77%)和人为干扰距离(1000m,76.92%)也较近(P0.05)。马麝春季生境变量的主成分分析结果表明,前4个主成分可以解释76.08%的数据差异,灌木因子(由灌木盖度组成)、乔木因子(由乔郁闭度和乔木密度组成)、食物因子(由坡位、乔木高度、食物多度组成)和隐蔽因子(由植被类型、隐蔽度组成)是影响马麝春季生境选择的重要因素。兴隆山马麝的生境选择是多维度进行的,是对其春季生境的食物可得性、隐蔽性、水资源及栖息基底等生态需求的综合适应。