焉耆马的DRA与DQA基因遗传多态性及其序列分析

采用聚合酶链式反应-单链构象多态性(PCR-SSCP)方法检测焉耆马ELA-DRA*exon2、ELADQA*exon2的基因型,并对不同等位基因进行测序,对其核苷酸与氨基酸序列进行分析。结果表明:焉耆马的ELA-DRA*exon2共检出4个等位基因;ELA-DQA*exon2共检出9个等位基因,通过核苷酸序列比对发现其中3个为新等位基因。ELA-DRA*exon2、ELA-DQA*exon2的各等位基因序列分析结果表明,发生非同义替换的多态位点均主要分布在抗原结合位点对应的编码序列中。遗传多态性分析结果表明,焉耆马的ELA-DRA*exon2多态信息含量为0.4,属于中度多态。卡方检验结果表明,焉耆马在该ELA-DRA*exon2位点处于Hardy-Weinberg平衡(χ2χ20.05)。本研究为马的抗病育种提供了理论基础。     

中国荷斯坦奶牛FASN基因部分序列多态性分析

利用PCR-SSCP技术,对中国荷斯坦奶牛脂肪酸合成酶(fatty acid synthase,FASN)基因第34外显子进行多态性分析。结果显示:该区域存在A、B两种等位基因和AA、BB、AB三种基因型,其中等位基因A和B的频率分别为0.890 8和0.109 2,基因型AA、BB和AB的频率分别为0.908 0、0.011 5和0.195 4。A为优势等位基因,AA为优势基因型。群体遗传学分析发现,荷斯坦奶牛FASN基因的有效等位基因数(Ne)、遗传杂合度(He)、遗传纯合度(Ho)和多态信息含量(PIC)分别为0.184 5、0.195 7、0.804 3和0.175 6。卡方检验表明,该群体已经极显著地偏离了Hardy-Weinberg平衡状态(P0.01)。说明中国荷斯坦奶牛FASN基因exon34区存在遗传多态性。     

荷斯坦牛TLR2基因遗传多态性与乳腺炎抗性关系分析

利用PCR-RFLP方法检测TLR2基因在297头中国荷斯坦牛中的多态性,并分析其多态性与体细胞评分(SCS)的相关性.结果显示,TLR2基因exon2扩增片断的109 bp处产生C→A的突变,并导致天冬氨酸改变为谷氨酸.TLR2基因exon2被EcoRV消化后表现多态性,表现AA、AB、BB 3种基因型,其中B等位基因为优势等住基因,等位基因频率为0.784 5,而A等位基因频率则为0.215 5.经X2适合性检验,中国荷斯坦牛在该位点达到Hardy-Weinberg平衡状态.同时,群体EcoRV基因座不同基因型与SCS相关分析的结果表明,BB、AB型个体的SCS指标显著高于AA型(P<0.05),AA基因型可作为乳腺炎抗性的有利基因型,可将TLR2作为奶牛乳腺炎候选基因,应用于乳腺炎抗性分子标记辅助选择育种.     

晋岚绒山羊多胎性能4个候选基因的研究

为探寻辅助育种的分子标记,提高绒山羊产羔数,本实验在山西省某绒山羊养殖场分别选取20只经产双羔母羊和单羔母羊,检测多胎性能候选基因GDF9、BMP15、FSHβ和GnRHR所有编码区SNPs位点;选取有差异的基因位点在171只经产母羊(90只双羔母羊和81只单羔母羊)中进行SNPs位点检测,统计基因型频率、等位基因频率、多态信息含量(PIC)和期望杂合度(He),并与母羊产羔数进行关联分析。结果显示:共发现17个SNPs位点,仅GDF9基因A959C位点和BMP15基因G735A位点在单、双羔母羊间有差异。SNaPshot检测表明GDF9基因A959C位点有AA、AC、CC 3种基因型,A为优势等位基因,AA为优势基因型,含CC、AC基因型个体的产羔数显著高于AA型。BMP15基因G735A位点共检测到GG、AG、AA 3种基因型,G为优势等位基因,GG为优势基因型;含AA、AG基因型个体的产羔数显著高于GG型。本研究结果表明GDF9基因A959C位点C等位基因和BMP15基因G735A位点A等位基因与晋岚绒山羊高产羔数呈极显著正相关,可作为分子标记对晋岚绒山羊进行辅助育种。     

哈萨克羊iNOS基因多态性与布鲁氏菌病的相关性分析

试验旨在探讨哈萨克羊诱导型一氧化氮合酶（iNOS）基因多态性与布鲁氏菌病的相关性。使用虎红平板凝集试验（RBPT）方法对231只哈萨克羊血清进行布鲁氏菌病血清学检测,参考GenBank中绵羊iNOS基因序列,针对其第6、7、8外显子及其邻近内含子片段设计引物,利用PCR-SSCP技术和DNA测序技术对231只哈萨克羊的iNOS基因进行多态性检测,分析其SNPs与哈萨克羊布鲁氏菌病易感性的相关性。结果表明,67只哈萨克羊为布鲁氏菌感染阳性,阳性检出率为29.00%。在哈萨克羊iNOS基因的外显子6和8片段上未检测到多态位点,在外显子7片段上检测出F7-T18054C和F7-C18084T 2个多态位点,在F7-T18054C多态位点上检测到3种基因型（TC、TT、CC）,优势等位基因和基因型分别是C型和CT型,其等位基因频率和基因型频率分别是0.660和0.446。在F7-C18084T多态位点上检测到2种基因型（CT、CC）,优势等位基因频率和基因型分别是C和CC型,其等位基因和基因型频率分别是0.946和0.892。F7-C18084T属于低度多态（PIC<0.25）,F7-T18054C属于中度多态（0.25 < PIC < 0.5）。相关性分析表明,F7-T18054C和F7-C18084T多态位点与布鲁氏菌病易感性无显著相关性（P>0.05）。试验结果表明,哈萨克羊iNOS基因F7-T18054C和F7-C18084T多态位点与布鲁氏菌病易感性不存在相关性。     

MyoG基因SNP检测及与藏羊体尺性状的关联性分析

利用PCR-SSCP及DNA测序技术,对632只河南县欧拉型藏羊、祁连县高原型藏羊和贵南县黑藏羊的MyoG基因exonⅠ多态性进行检测并分析多态位点与体尺性状的相关性。结果表明,MyoG基因exonⅠ109位存在A→C的碱基突变,存在CC、CA、AA3种基因型,该突变导致MyoG基因exonⅠ37号位谷氨酸(GAG)→丙氨酸(GCG)的错义突变;该位点在3个藏羊群体中均处于中度多态;最小二乘法分析表明,河南县欧拉羊群体中CC基因型个体的体重、体高和体长均显著高于AA基因型个体(P0.05),祁连县高原型藏羊群体中CC基因型个体的体重显著高于AA基因型个体(P0.05),贵南县黑藏羊群体中CC基因型个体的体长显著小于CA基因型和AA基因型个体(P0.05)。     

通过PCR-SSCP检测乌珠穆沁马ELA-DQA第二外显子的多态性

实验通过PCR-SSCP技术检测了50匹中国乌珠穆沁马的ELA-DQA第二外显子。结果表明:50匹乌珠穆沁马中共出现11种基因型:5种纯合子分别记为AA、BB、CC、DD和EE型,其中AA型为单带;其余4种纯合子均为3条带;6种杂合子分别记为AB、AC、AD、AE、BD和BE型,带型不一,其中A、B、C、D和E等位基因频率分别为0.28、0.25、0.11、0.16和0.20。这表明乌珠穆沁马ELA-DQA第二外显子多态性丰富。     

绵羊MyoG基因外显子的单核苷酸多态性群体遗传学分析

采用PCR—SSCP技术分析了肌细胞生成素（MyoG）基因外显子1、2在蒙古羊、无角陶赛特羊、德国肉用美利奴羊和白萨福克羊这4个绵羊品种的多态性。结果表明在在外显子1（exonⅠ）所扩增的片段中存在3种基因型（AA型、AB型和BB型），在外显子2（exonⅡ）所扩增的片段中不存在多态性。对于exonⅠ扩增片段，4个绵羊品种均检测到AA和AB基因型；BB基因型只在蒙古羊、陶赛特羊和白萨福克羊中检测到。在4个绵羊品种中，蒙古羊的AA基因型频率最高，而其它3个品种羊是AB基因型频率高；A等位基因频率明显高于B等位基因频率。exonⅠ的多态性片段测序分析表明：MyoG基因第305处发生了单碱基突变（T→C），并导致了所编码氨基酸由半胱氨酸变为精氨酸。     

绵羊GLP2R基因多态性及其与肉质性状的相关分析

为探索胰高血糖素样肽-2受体(glucagon-like peptide-2 receptor,GLP2R)基因多态性与绵羊肉质性状的相关性,试验选用68只南非肉用美利奴羊×东北细毛羊和102只杜泊羊×小尾寒羊的杂交后代为研究对象,采用Sanger测序方法寻找GLP2R基因外显子11序列的突变位点,根据测序结果分析该突变位点的基因型频率和基因频率及卡方适合性检验、纯合度(Ho)、杂合度(He)和多态信息含量(PIC)。结果显示,绵羊GLR2R基因外显子11处存在C/T突变,存在3种基因型:CC、TC和TT,2种等位基因:C和T,其中在南非肉用美利奴羊×东北细毛羊群体中,TC基因型为优势基因型,在杜泊羊×小尾寒羊群体中,CC基因型为优势基因型;2个群体中C均为优势等位基因。卡方适合性检验结果显示,2个群体的C/T位点均处于Hardy-Weinberg平衡状态(P0.05);C/T突变位点在群体内变异较小且处于中度多态(0.25PIC0.5),有一定遗传学意义。不同基因型肉质性状差异结果显示,南非肉用美利奴羊×东北细毛羊TC基因型剪切力显著高于CC和TT基因型,TT基因型失水率显著高于CC和TC基因型(P0.05);杜泊羊×小尾寒羊CC基因型剪切力显著高于TC和TT基因型,TT基因型失水率显著高于CC和TC基因型(P0.05)。本研究在GLP2R基因外显子处发现C/T突变,可能是影响肉质性状的潜在位点,是否能够作为遗传标记还需要进一步探索。     

甘肃马鹿MyoG基因5′UTR单核苷酸多态性检测及其序列变异分析

为了初步探索肌细胞生成素(MyoG)基因在鹿科动物上的遗传多态性,试验采集56头甘肃马鹿的血液样品,采用PCR-SSCP和克隆测序的方法对MyoG基因(GenBank登录号：FJ746497)5′UTR部分序列进行了单核苷酸多态性(SNPs)检测及序列变异分析。结果表明：①引物P1的PCR扩增片段存在多态性,经克隆测序分析,在5′UTR -237 bp处存在G→A的转换变异,该位点表现为AA、AB和BB 3种基因型,由A和B 2个等位基因控制;对基因型、等位基因频率及群体遗传多态性分析结果发现,AA基因型频率最高,A基因为优势等位基因,该位点多态信息含量(PIC=0.194)为低度多态(PIC＜0.25);经TFSEARCH 1.3分析软件预测发现,该变异可能减少了1个Sp1转录因子。②引物2的PCR扩增片段检测到1个SNP多态位点(—46 bp,G→A),表现为CC、CD和DD 3种基因型, 基因型频率分布为CC＞CD＞DD,该位点多态信息含量(PIC=0.266)为中度多态(0.25＜PIC＜0.5),但该变异并未引起潜在调控元件及蛋白质结合位点的改变。本研究结果为进一步分析MyoG基因SNPs位点与甘肃马鹿胴体、肉质性状的关联性奠定了基础。     

马生长激素基因多态性与体尺指标之间的关联性分析

为研究广西德保矮马、百色马和新疆伊犁马的生长激素(GH)基因多态性与体尺指标之间的相关性,本实验利用PCR-SSCP技术分析3个品种277匹马GH基因5′侧翼区、第3外显子、第4外显子、第5外显子的遗传多态性。结果表明:只有GH基因第5外显子出现多态性,表现为AA、BB和AB 3种基因型,其他片段未发现多态性。群体遗传学分析表明,等位基因A和B的基因频率相等;Hardy-Weinberg平衡检验显示,3个品种马均处于非平衡状态;它们的多态信息含量均为中度多态。统计分析表明,GH基因第5外显子的基因型与3个品种马体尺指标之间无显著相关性。     

ELA-DRA polymorphisms are not associated with Equine Arteritis Virus infection in horses from Argentina

Polymorphisms at Major Histocompatibility Complex (MHC) genes have been associated with resistance/susceptibility to infectious diseases in domestic animals. The aim of this investigation was to evaluate whether polymorphisms of the DRA gene the Equine Lymphocyte Antigen is associated with susceptibility to Equine Arteritis Virus (EAV) infection in horses in Argentina. The equine DRA gene was screened for polymorphisms using Pyrosequencing? Technology which allowed the detection of three ELA-DRA exon 2 alleles. Neither allele frequencies nor genotypic differentiation exhibited any statistically significant (P-values=0.788 and 0.745) differences between the EAV-infected and no-infected horses. Fisher's exact test and OR calculations did not show any significant association. As a consequence, no association could be established between the serological condition and ELA-DRA.     

玉树马和德保矮马SLC36A1基因多态性研究

 控制马香槟毛色的CH（Champagne gene）基因家族包含4个候选基因（SLC36A1、SLC36A2、SLC36A3、SPARC）,研究发现SLC36A1基因外显子2的突变是造成马香槟毛色的关键位点。为揭示中国马SLC36A1基因遗传多态性,本研究以玉树马和德保矮马共74个样本为研究对象,以马DNA池为模板扩增SLC36A1基因的10个外显子及部分内含子序列并进行测序分析。共发现马SLC36A1基因5个SNPs,分别位于内含子3（g.26699953 A>G, g.26699851 G>C, g.26699850 G>C）,外显子4（g.26699562 G>A）及外显子6（g.26697018 C>T）。利用PCR RFLP方法对74个家马样本进行基因分型,发现外显子6的SNP有基因型CC、CT;外显子4的SNP有基因型GG、GA;内含子3的g.26699850 G>C突变有基因型GG、GC;内含子3的另外2个SNPs（g.26699953 A>G, g.26699851 G>C）通过测序,发现有AA、AG、GG与GG、GC、CC基因型。所有5个马SNPs均为野生型占主要优势。由此界定了马SLC36A1基因有9种单倍型（H1 H9）,其中H5是最主要的单倍型。德保矮马遗传多样度为0.4190,比玉树马（0.2228）高,表明德保矮马香槟毛色遗传多态性比玉树马更丰富。玉树马与德保矮马的平均单倍型多样度为0.3160,表明其香槟毛色遗传多态性相对较低。     

哈萨克马和焉耆马不同组织中IGFBP-5基因表达差异研究

试验旨在探求胰岛素样生长因子结合蛋白-5（insulin-like growth factor binding protein-5,IGFBP-5）基因在哈萨克马和焉耆马不同组织部位中的mRNA表达规律。采用实时荧光定量PCR技术检测在哈萨克马和焉耆马心脏、肝脏、脾脏、肺脏、肾脏、小肠、大肠、盲肠、肋间肌、背最长肌、臂肌和臀肌等不同组织中IGFBP-5基因mRNA表达量,同时比较该基因在哈萨克马和焉耆马相同组织中的表达差异。结果表明,哈萨克马和焉耆马表达量最高的均为背最长肌,其次是臂肌,并显著高于心脏、肝脏、脾脏、肺脏、肾脏、小肠、大肠、盲肠等内脏组织表达量（P<0.05）,大肠中的表达量最低;焉耆马的肾脏、小肠、大肠、盲肠、背最长肌和臂肌中表达量高于哈萨克马相同部位的表达量,其中焉耆马背最长肌和大肠中的表达量极显著高于哈萨克马的相同部位（P<0.01）,小肠和盲肠中的表达量显著高于哈萨克马的相同部位（P<0.05）。本试验为深入研究IGFBP-5基因的生物学功能,以及对中国马生产性能的遗传改良提供理论依据。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

Kinematic characteristics of Andalusian, Arabian and Anglo-Arabian horses: a comparative study

The aim of this study was to compare the kinematic trot characteristics of three different breeds of horse: Andalusian (AN, n = 15), Arabian (AR, n = 7) and Anglo-Arabian (AA, n = 5) using standard computer-assisted videography (25 Hz). Linear, temporal and angular parameters in fore- and hind limbs were analysed in six randomly selected strides per horse. Normalised angle-time diagrams along the complete stride were obtained for all joints angles in each breed and specific kinematic characteristics were detected graphically. AA horses displayed longer swing durations in both limbs ans a shorter angular range of motion (ARM) in scapula and pelvis inclination and in shoulder, hip and forelimb retraction-protraction angles. At lift off, stifle and tarsal joint angles were more flexed. In general, only small differences were observed in AR horse kinematics when compared with the other 2 breeds. AN horses presented negative overtracking length, which was positive in AR and AA. In AN horses the elbow and carpal joints were more flexed at the moment of maximal elevation, elbow and fore-fetlock joints also exhibited a larger ARM due to a smaller angle at maximal flexion. In the hind limbs, tarsal, hind fetlock and retraction-protraction angles presented a larger ARM in AN horses due to greater maximal flexion in the tarsal and hind fetlock joints. Fore- and hind fetlocks were also more flexed in horses from this breed. In conclusion, differences between kinematic variables at the trot were observed in the three breeds studied here, mainly in forelimb joints. The most outstanding feature was the greater forelimb flexion recorded in AN horses than in the other breeds which is consistent with the elevated movements in this breed. In AA horses, the ARM of proximal joints involved in retraction protraction in both fore- and hind limbs was smaller. All the differences observed highlighted the idiosyncratic nature of the trot in each breed; this may influence the functional capacity of each breed.     

不同品种马ELA-DQA* exon2的多态性分析

通过PCR—SSCP技术检测了7个品种300匹马ELA—DQA＊^exon2的多态性。用12％非变性聚丙烯酰胺凝胶电泳将已变性的PCR扩增产物分离，银染法显色。结果300匹马中共出现27种基因型：10种纯合子和17种杂合子；7个品种马的He和PIC值均表现为高度多态；等位基因序列分析得到10个等位基因间的Kimura双参数遗传距离为0．015～O．147，符合（dN：〉dS）的正选择检验。因此表明马的ELA—DQA＊^exon2多态性丰富。     

新疆阿勒泰地区哈萨克马乳房参数与乳房形状调查

为了确定新疆阿勒泰地区哈萨克马的乳用优势,本研究测量了131匹阿勒泰地区成年泌乳高峰期哈萨克马乳房外部结构,总结其特征并测定了挤奶相关参数。结果显示,阿勒泰地区哈萨克马乳房周长平均为58.48 cm,乳房深度为13.24 cm,乳房沟长为24.70 cm,乳头长为4.27 cm,乳头根周长为11.13 cm,两个乳头间距为5.43cm。阿勒泰地区哈萨克马的乳房结构参数平均值接近碗装乳房,其中碗状乳房比率在52.67%,球状乳房比率在34.35%,梨状乳房比率在12.98%,这个比例说明阿勒泰地区哈萨克马具有较大的乳用潜力。同时计时方法测定结果为人工挤奶127.21次/min,马驹吮吸112.4次/min,脉动35.64次/min,可在此基础上优化人工挤奶次数,为马用挤奶器的开发提供数据支持。     

Prevalence of behavioral disorders in the Swiss horse population

In the Swiss horse population, the prevalence of conspicuous behaviours (behavioural disorders and stable vices) was recorded with a written questionnaire in a representative survey among 1861 horse yards, and the occurrence of the stereotypic behavioural disorders crib-biting, weaving and boxwalking was analysed with emphasis on their association with horse specific (breed, age, sex) and environmental factors (e.g. housing system, nutrition, management and utilisation). 622 horse yards with a total number of 2536 horses answered to the questionnaire, and conspicuous behaviours were described for 418 horses (16.5%). Thereof, the most common stereotypic behavioural disorders with a potential negative impact on animal welfare, crib-biting, weaving, and/or boxwalking were named for 89 horses (3.5%). Stable vices (e.g. shying, bucking, aggressivity against humans) were noted for 47 horses (1.9%). For 281 horses (11.1%) conspicuous behaviours such as pawing, lip-liking, head shacking, kicking stall were given; these can be categorised as behavioural disorders or as stable vices depending on their intensity and causation. Data analysis data revealed a number of statistically significant associations between genetic factors (breed), housing, management practices, utilisation and the three stereotypies. The strongest influence on the degree of behavioural disorders had the breed as well as the extent of direct social contact with other horses, free movement on pasture, feeding pattern and regular utilisation.     

Effect of difference in training skills on stress in horses trained by Kazakh trainers

This study aimed to evaluate the effect of differences in trainer skills on horse training during the early stages of riding habituation by measuring the levels of stress and changes in stress levels. Among nine untrained horses employed, five in Group A were trained by two low-skilled trainers, whereas the remaining four in Group B were trained by two high-skilled trainers using the traditional Kazakh method. Salivary α-amylase concentration was measured as a biomarker of stress immediately before and after each riding session during the training period. In the duration of riding and mooring times to the total riding habituation time for each horse, no significant difference was observed between the two groups. In contrast, the mean total stress and mean final stress were significantly lower in Group B than in Group A, and the mean total change in stress before and after riding habituation was significantly higher in Group B. Differences in trainer skills were evidenced as differences in training methods to suppress the total stress levels through differences in the application of stress burden during the training of individual horses.