东北梅花鹿优化育种规划中育种目标的确定

讨论了东北梅花鹿依据经济指标确定育种目标的问题 ,以及在东北梅花鹿育种规划中 ,应如何挑选在综合育种目标中的生产性状及其对应的选择性状。并着重介绍了应用综合育种值作为数量化育种目标的方法。指出在确定以茸用为主的东北梅花鹿育种目标时 ,除了主要考虑茸用性状外 ,还应适当地考虑繁殖和使用寿命等性状 ,同时对于直接影响茸鹿生产效益的直接性状和次级性状不容忽视。     

The Construction of Cloned Sika Deer Embryos (Cervus nippon hortulorum) by Demecolcine Auxiliary Enucleation

The objective of our study was to establish the feasibility of experimental protocols for cloning sika deer. We performed auxiliary enucleation to improve the efficiency of nuclear transfer operation by optimizing the demecolcine concentration to induce cytoplasmic protrusions in the sika deer oocytes. In the present study,we had studied the impact of different demecolcine concentrations on cytoplasmic protrusions and enucleation rates. We determined that 95.9% of the sika deer oocytes formed cytoplasmic protrusions when treated for 1 h with 0.8 μg/ml demecolcine. The lowest observed rate of protrusion was 19.3% after overnight treatment with demecolcine. When the oocytes aged or had a poor cumulus expansion, they exhibited a significant decrease in the ability to form cytoplasmic protrusions. The rates of enucleation (94.9% vs 85.8%, p < 0.05), cell fusion (84.6% vs 70.1%, p < 0.05) and blastocyst formation (15.4% vs 10.9%, p < 0.05) using demecolcine auxiliary enucleation were significantly higher than those after blind enucleation. These results demonstrated that sika deer oocytes could be enucleated quickly and effectively using demecolcine auxiliary enucleation, which could enhance the enucleation rate, cell fusion rate and blastocyst rate of cloned embryos in vitro.     

梅花鹿三种保定药物对比实验

本文通过用静松灵、司可林、眠乃宁三种保定药物保定梅花鹿对比实验，得出：①静松灵一次性给药剂量较大，生产中操作不方便。②司可林一次给药剂量虽小，但有效剂量与致死剂量十分接近，剂量误差易导致鹿死亡。③静松灵和司可林均无有效拮抗药物，且后遗性影响较大。④眠乃宁注射剂量适中，剂量误差不会导致鹿死亡，无后遗性影响。拮抗药物苏醒灵拮抗效果优良，使眠乃宁的应用更安全。     

全国梅花鹿养殖状况调查

中国野生动物保护协会于2005年6～12月,在全国31个省(区、市)开展了梅花鹿养殖情况调查.调查数据显示,截止到2004年底,全国31个省(区、市)共有9 465家养殖场,圈养繁殖梅花鹿452 355头.其中养殖梅花鹿数量最多的是吉林省,养殖场5 985家,目前存栏量278 000头.2004年全国生产梅花鹿鹿茸73 t,吉林省居全国首位,年产量约为21 t,占总数的28.8%.本文以调查数据为基础,提出我国梅花鹿养殖中存在的问题和改进建议.     

梅花鹿感染产气荚膜梭菌的诊断及中西医结合治疗

2020年5月25日,贵州省某动物园的梅花鹿(Cervus nippon)出现精神沉郁、食欲减退、腹部胀大等症状,截至5月29日已发病死亡6头。为查明发病原因,采集发病梅花鹿的血样及发病死亡梅花鹿的组织样品送检。根据临床症状和病理剖检结果,初步诊断该动物园发病梅花鹿疑似产气荚膜梭菌(Clostridium perfringens)和支原体混合感染,经细菌分离培养、生化特性鉴定确诊为产气荚膜梭菌感染。结果显示,造成该动物园梅花鹿发病死亡的原因为产气荚膜梭菌的感染,并根据其症状进行中西医结合治疗。     

梅花鹿朊蛋白基因(PRNP)的克隆及序列分析

根据GenBank中鹿朊蛋白基因序列设计特异性扩增引物，采用PCR方法从中国梅花鹿基因组中扩增得到梅花鹿的朊蛋白基因，采用PCR产物直接测序，并将其克隆到pGEM—TEasy载体中测序进行进一步确认，通过分析表明所克隆的梅花鹿朊蛋白基因的ORF基因片段包含771bp，编码256个氨基酸的前体蛋白，相对分子质量约为28200。与已报道的其他品种鹿朊蛋白基因序列进行对比分析，核苷酸及氨基酸序列的同源性均在98％以上。本试验为进一步研究中国梅花鹿朊蛋白的多态性提供了数据。     

梅花鹿瘤胃纤维素降解菌的分离鉴定及其酶活力测定

为获得来源于梅花鹿瘤胃的高效纤维素降解菌,本试验采集了3锯龄健康梅花鹿的新鲜瘤胃液,用羧甲基纤维素钠（CMC-Na）培养基分离筛选纤维素降解菌,综合其形态学、生理生化特征和16S rDNA基因测序等对分离菌株进行分类学鉴定,利用DNS法对菌株的产纤维素酶条件进行初步研究,并探讨以不同物质为底物时的纤维素酶酶学特性,为后期菌株的应用提供理论数据。结果显示,本试验从梅花鹿瘤胃液中分离的菌株N-11是一株高效纤维素降解菌,经鉴定为蜡样芽孢杆菌（Bacillus cereus）。产酶性质研究结果表明,该菌株产纤维素酶活力在30~45℃（最适温度为40℃）,pH为6.0~7.0（最适pH 6.0）,碳源含量为2%~5%（最佳接种量5%）较高;培养36 h时达到产酶高峰,纤维素酶酶活力（CMCA）和滤纸酶酶活力（FPA）分别为12.563、12.414 U/mL,在20~50℃或pH 6.0~8.0环境下作用1 h,其相对酶活力均保持在80%以上,稳定性较好。以上结果表明,蜡样芽孢杆菌N-11具有较高的纤维素酶活力,在纤维素利用方面具有较高的应用价值。     

游客密度对圈养梅花鹿行为影响

2012年10月1～6日和18 ～23日,以昆明动物园内的圈养梅花鹿为研究对象,采用目标动物取样法和全事件记录法,研究了不同游客密度条件下梅花鹿的行为差异并探讨了游客密度对梅花鹿日常行为的影响.调查结果显示,不同游客密度对梅花鹿的移动和观望行为影响显著,雄鹿受影响最显著的行为是观望和其他行为(P＜0.01),其次是移动和社会行为(P＜0.05);雌鹿受游客密度影响最显著,观望、取食和反刍行为均有极显著差异(P＜0.01),其次是移动行为(P＜0.05);而幼鹿受影响最显著的行为是取食行为(P＜0.01),其次是移动、观望和反刍行为(P＜0.05).建议:(1)改变投食时间;(2)在繁殖期将雌鹿隔离到游客干扰小的区域饲养;(3)增加笼舍内的植被覆盖度及退避区域,减少动物在与游客接触过程中的应激性.     

梅花鹿(Cervus nippon)保护遗传学研究现状及其保护愿景展望

梅花鹿(Cervus nippon)为国家Ⅰ级重点保护野生动物,分化为6个亚种:东北亚种(C.n.hortulorum)、华北亚种(C.n.mandurinui)、华南亚种(C.n.kopschi)、四川亚种(C.n.sichunicus)、山西亚种(C.n.grassianus)、台湾亚种(C.n.taiouanus),其中山西亚种、华北亚种和台湾亚种野外种群已经灭绝,野外残存的东北亚种、华南亚种和四川亚种仅分布于黑龙江、吉林、江西、浙江、安徽、四川和甘肃等省破碎的栖息生境内。由于残存的3个亚种种群数量稀少,野外很难发现,这对保护和恢复濒临灭绝的野生梅花鹿种群造成一定的困难。中国对圈养梅花鹿生物学的报道虽然较多,但是在行为学、遗传学方面的研究较少,且分子标记方面的研究也主要集中在与经济性状相关的基因方面。为了更好地保护和恢复野生梅花鹿种群,本文在查阅了相关文献的基础上,概述了国内外梅花鹿遗传学研究现状与进展,展望了野生梅花鹿需要进一步研究的趋势,以期为我国野生梅花鹿的保护管理和种群恢复提供科学依据。     

梅花鹿行为生态学及保护遗传学研究进展

梅花鹿是我国极度濒危的鹿科动物,目前仅分布于黑龙江、吉林、四川、江西等省狭窄的区域内。文章主要对梅花鹿的种群数量、行为生态学及保护遗传学研究等进行了概述,并指出目前国内学者对梅花鹿行为生态学的研究主要集中在行为节律和栖息地的研究,为了更准确地反映梅花鹿系统进化关系,必须结合形态学、动物地理学、地理信息系统及古生物学等方面进行综合分析探讨。     

梅花鹿朊蛋白核心片段的基因克隆与高效表达

本试验根据梅花鹿朊蛋白基因序列设计引物,利用PCR的方法从梅花鹿基因组DNA中扩增朊病毒蛋白酶抵抗区域PrP^res,将该片段分别与表达载体pET-Trx和pET-His连接,构建重组表达载体pET-Trx-PrP^res和pET-His-PrP^res。分别将两个重组表达载体转入E.coli BL21(DE3) plys宿主菌中,37 ℃诱导4 h,经SDS-PAGE分析,Trx-PrP^res和His-PrP^res表达量分别为38.2%和30.1%。     

山羊卵母细胞体外成熟的研究

在38℃和39℃不同温度条件下培养山羊卵母细胞,结果表明:在38℃和39℃下成熟率分别为63.6%和57.3%,二者之间差异不显著,说明只要培养温度在山羊正常体温的变动范围内,对其卵母细胞的成熟率没有影响;在38℃温度条件下用10?S的M199培养液成熟率为62.5%,而10 mg/ml BSA的M199培养液成熟率为58.7%,二者之间差异不显著,说明无血清培养可以代替血清培养.     

不同形式铜对雄性梅花鹿血清生化指标及营养物质消化率的影响

本文旨在研究不同形式铜对雄性梅花鹿血清生化指标及营养物质消化率的影响,进而筛选出在梅花鹿日粮中铜的最适宜添加形式.选取20头2年生梅花鹿随机分成4组(A组、B组、C组和D组),按照能量相近、营养物质一致、铜添加量均为10 mg/kg(铜离子)的基础下,研究硫酸铜(B组)、柠檬酸铜(C组)和蛋氨酸铜(D组)3种铜源对雄性梅花鹿血清生化指标及营养物质消化率的影响.试验结果表明:1)蛋氨酸螫合铜组对干物质、粗蛋白质及钙的消化率高于另外3组(P>0.05);蛋氨酸螯合铜组对磷的消化率极显著高于其对照组和硫酸铜组(P<0.01);2)蛋氨酸铜组血清铜含量极显著高于另外3组(P<0.01),毛中铜含量高于基础日粮组和硫酸铜组(P<0.05),粪中铜含量低于硫酸铜组和柠檬酸铜组(P<0.01);3)不同铜源影响总蛋白质、白蛋白、碱性磷酸酶、谷丙转氨酶、谷草转氨酶活性,但各组间差异不显著(P>0.05),各组间血糖含量差异不显著(P>0.05);4)蛋氨酸铜组铜蓝蛋白活性、超氧化物歧化酶活性显著高于基础日粮组(P<0.05).结果表明,梅花鹿饲粮中最适宜的添加铜源为蛋氨酸铜.     

海狸鼠卵母细胞体外成熟培养初探

实验用ＰＭＳＧ或ＰＭＳＧ＋ＨＣＧ处理或未经激素处理的海狸鼠８只，共获卵巢卵母细胞１３８枚。激素处理对获取卵巢卵母细胞的数量没有影响，而对体外成熟发育至卵丘扩展和半成熟阶段有促进作用。三种不同培养液（Ｗｈｉｔｅｎ＋ＦＣＳ；ＴＣＭ１９９＋ＰＭＳＧ＋ＦＣＳ；ＴＣＭ１９９＋ＨＣＧ＋ＦＣＳ）共培养１２５枚卵母细胞，培养后卵丘扩展率及半成熟率分别为５６．５％，４５．７％，４７．６％和２１．７％，１２．３％，９．５％，以Ｗｈｉｔｅｎ液较高（分别为５６．５％和２１．７％），但只有ＴＣＭ１９９＋ＰＭＳＧ＋ＦＣＳ组有２枚卵母细胞出现第一极体。结果表明海狸鼠卵母细胞与其它啮齿动物的卵母细胞一样，能够在体外培养成熟，完成第一次减数分裂，排出第一极体     

Ghrelin Accelerates In Vitro Maturation of Bovine Oocytes

Ghrelin, apart from its metabolic role, is nowadays considered as a basic regulator of reproductive functions of mammals, acting at central and gonadal levels. Here, we investigated for possible direct actions of ghrelin on in vitro maturation of bovine oocytes and for its effects on blastocyst yield and quality. In experiment 1, cumulus oocyte complexes (COCs) were matured in the presence of four different concentrations of ghrelin (0, 200, 800 and 2000 pg/ml). In vitro fertilization and embryo culture were carried out in the absence of ghrelin, and blastocyst formation rates were examined on days 7, 8 and 9. In experiment 2, only the 800 pg/ml dose of ghrelin was used. Four groups of COCs were matured for 18 or 24 h (C18, Ghr18, C24 and Ghr24), and subsequently, they were examined for oocyte nuclear maturation and cumulus layer expansion; blastocysts were produced as in experiment 1. The relative mRNA abundance of various genes related to metabolism, oxidation, developmental competence and apoptosis was examined in snap‐frozen cumulus cells, oocytes and day‐7 blastocysts. In experiment 1, ghrelin significantly suppressed blastocyst formation rates. In experiment 2, more ghrelin‐treated oocytes matured for 18 h reached MII compared with controls, while no difference was observed when maturation lasted for 24 h. At 18 and 24 h, the cumulus layer was more expanded in ghrelin‐treated COCs than in the controls. The blastocyst formation rate was higher in Ghr18 (27.7 ± 2.4%) compared with Ghr24 (17.5 ± 2.4%). Differences were detected in various genes’ expression, indicating that in the presence of ghrelin, incubation of COCs for 24 h caused over‐maturation (induced ageing) of oocytes, but formed blastocysts had a higher hatching rate compared with the controls. We infer that ghrelin exerts a specific and direct role on the oocyte, accelerating its maturational process.     

In Vitro Maturation of Oocytes from Norwegian Semi-domestic Reindeer (Rangifer tarandus)

In vitro maturation of oocytes has been described for several species, but to our knowledge this is the first attempt to mature oocytes from reindeer in vitro. Reindeer are seasonal breeders with their main period of breeding activity starting in the beginning of October. Little knowledge exists about in vivo oocyte maturation, fertilization and development of the early embryonic stages after fertilization. The aim of the present study was to examine whether reindeer oocytes collected out of breeding season could be matured in vitro in conventional medias used for bovine oocytes.     

Ultrastructure of aortic elastic fibers in copper-deficient Sika deer (Cervus nippon Temminck)

Light microscopic and transmission and scanning electron microscopic observations were performed on the aortas of two 4- and 6-year-old deer affected with cervine ataxia and two 6-month- and 4-year-old healthy deer. Examination of the aortas from affected deer by transmission electron microscopy revealed the absence of distinct elastic laminae in the internal elastic lamina and tunica media, but discontinuous and irregular clumps of elastin were present. Scanning electron microscopy disclosed immature architecture of elastic fibers in the aortas from the copper-deficient deer, and the architecture was similar to that of a 6-month-old healthy deer.