西藏牦牛和黑白花奶牛生长激素基因Alu I多态性的比较研究

利用聚合酶链式反应（PCR）技术，测定了西藏牦牛和黑白花奶牛生长激素基因的Alu I-RFLP（限制性内切酶片段长度多态性），并比较了2牛群的基因频率。在西藏牦牛和黑白花奶牛中，等位基因A的频率分别为0.935和0.905，等位基因B的频率分别为0.065和0.065，卡方检验差异不显著（P＞0.05）。     

牦牛生长激素基因的测序和多态性研究

采用PCR产物直接测序法获得牦牛生产激素基因（GH基因）891bp序列。通过GenBank中的Blastn软件进行序列比较分析表明，牦牛GH基因与普通牛GH基因的同源性为98%，与水牛，山羊，绵羊同源性分别为96%，94%，93%：NJ法构建的分子系统树显示，牦牛与普通牛亲缘关系最近，其次为水牛，而与山羊，绵羊的亲缘关系较远，此外，采用PCR-RFLP方法研究了30头牦牛GH基因891bpDNA片段，未发现MspⅠ/HpaⅡ酶切位点多态性，表明牦牛GH基因的多态性比较贫乏。     

青海黑白花奶牛乳蛋白的多态性

采用聚丙烯酰胺凝胶电泳法对西宁市奶牛场自繁的272头青海黑白花泌乳奶牛乳样的四个乳蛋白多态性系统进行了研究。结果发现α-乳白蛋白位点只有α-LaBB 型,β-乳球蛋白位点有β-LgAA、β-LgAB 和β-LgBB 三型,α_(S1)-酪蛋白位点有α_(S1)-CnBB 和α_(S1)-CnBC 两型,β-酪蛋白位点有β-CnAA 和β-CnAB 两型,同时还对青海黑白花奶牛两个群体之间每一个等位基因的固定指数、遗传相似性和遗传距离进行了讨论。     

牦牛乳及乳制品、犏牛和黑白花奶牛乳的脂肪酸组成分析

用气相色谱 质谱联用仪(GC MS)对牦牛乳、黑白花奶牛乳、犏牛乳及牦牛乳制品（奶油、酥油、曲拉、酸奶）中的脂肪酸组成进行了测定。结果表明,牦牛乳中功能性脂肪酸,如共轭亚油酸（CLA）、亚油酸（LA）、α 亚麻酸（ALA）、γ 亚麻酸（GLA）占总脂肪酸的比重均显著高于犏牛乳和黑白花奶牛乳（P＜0.05）;犏牛乳中ω 6/ω 3 PUFA的比值（1.55）略高于牦牛乳（1.54）,差异不显著（P＞0.05）,但都在最佳膳食平衡比值范围内,黑白花奶牛乳中ω 6/ω 3多不饱和脂肪酸（PUFA）的比值（11.33）超过了推荐最佳比值。加工处理能够改变乳制品脂肪酸的构成,如牦牛乳奶油中检测出原奶中所不含的一种亚油酸（18:2Δ8c,11c）。酥油主要以不饱和脂肪酸（UFA）为主,而曲拉主要以饱和脂肪酸（SFA）为主,牦牛酸奶中没有检测到GLA。     

牦牛与黑白花牛的氮平衡比较研究

接近初情期的牦牛和黑白花公牛各3头，于代谢厩中进行精料型（精粗比为4：1）和粗料型（精粗比为1：4）日粮条件下的氮平均对比试验。结果表明，不论牦牛还是黑白花牛，干物质采食量（DMI）、粪氮（FN）、尿氮（UN）和干物质消化率（DMD）均随精料含量上升而升高（P＜0．01），而进食氮（NI）的利用效率则随NI的升高而下降（P＜0．01）。在粗料型的日粮条件下，牦牛对NI的利用效率（58．2％）高于（P＜0．05）黑白花牛（49．0％），二者的DMD（45％）无差异；而在精料型的日粮条件下，NI的利用效率（22％）和DMD（64％）均不随牛种而变化（P＞0．05）。从这些结果得出结论：牦牛与黑白花牛消化道的吸收效率是一样的，在精料型日粮条件下，牦牛机体组织存留氮的能力低于黑白花牛；在粗料型日粮条件下，牦牛比黑白花牛更能有效地利用蛋白资源。     

新西兰黑白花奶牛适应性的研究

<正> 四川省绵阳市于1986年1月引进新西兰黑白花奶牛176头(以下简称新牛),为了养好和充分利用这批奶牛,我们结合生产,以新牛为试验组,用本地黑白花奶牛(以下简称本牛)作对照,进行了适应性的研究,现将结果简报于后。     

牦牛的黑白花牛瘤胃内饲料蛋白质降解率的比较研究

用瘤胃尼龙袋技术研究了青藏高原８种常见饲料在牦牛和黑白花牛瘤胃内的粗蛋白（ＣＰ）和干物质（ＤＭ）降解率,分析了２个动物品种间的降解率的差异性。装有永久性瘤胃瘘管的牦牛（平均体重１００ｋｇ）和黑白花牛（平均体重１７０ｋｇ）各２头,以１．５×ＭＥｍ水平饲喂同一基础日粮（精粗比为５０：５０）,瘤胃食糜的外流速度估计为０．０６７／ｈ（精料）和０．０２／ｈ（粗料）。待测料在瘤胃内的培养时间为２ｈ、４ｈ、６ｈ     

国际黑白花奶牛品系比较（一）

为提高波兰黑白花奶牛的生产性能,联合国粮农组织从1974年至1984年引用10个国家的黑白花奶牛(弗里生、荷兰—弗里生)冷冻精液与波兰黑白花奶牛杂交,进行杂交后裔的生产性能比较试验,意欲选择理想的杂交组合,并建立种牛群。同时希望取得试验的结果。有助於一些国家在应用杂交育种途径,提高当地牛的产奶量和产肉量的试验     

奶牛群体生长激素基因多态性与产奶量关联性

设计7对引物,利用PCR-SSCP技术检测GH基因在130头中国荷斯坦奶牛群体的遗传多态性,研究该基因对产奶量的影响。结果发现在GH基因的第3内含子、第5外显子和3′端存在多态性。GH基因第3内含子等位基因A、B的基因频率为0.775和0.225,3′端等位基C、D在奶牛群体中的基因频率为0.729和0.271,GH基因第5外显子等位基因E、F和G、H在奶牛群体中的基因频率分别为:0.507、0.138和0.259、0.096,GH基因第3内含子座位上不同基因型对305d产奶量有显著效应,AB基因型305d产奶量显著高于AA和BB型(P0.05)。     

国际黑白花奶牛品系比较（续完）

二、母牛第一胎生产纪录: 1、初产日龄及体重青年母牛初产日龄,放牧的F_1平均为885又,R_1为916天。集约化饲养的稍早,分别为870及837天。初产犊后平均体重,放牧的F_1为493公斤,R_1为476公斤。集约化饲养的,F_1为475公斤,R_1为463公斤。详见表5。     

三个牦牛群体DRB3.2基因PCR-RFLP多态性研究

为了研究牦牛DRB3.2基因exon 2遗传多样性,试验采用PCR-RFLP对天祝白牦牛、甘南牦牛、大通牦牛3个类群757头个体的MHC-DRB3.2基因进行PCR-RFLP分析。结果表明:共检测出8个HaeⅢ酶切位点、11种基因型;在3个牦牛类群中,HaeⅢC基因型(225 bp/175 bp/85 bp/35 bp)在大通牦牛、天祝白牦牛中是优势基因型,基因型频率分别为0.387和0.366;而在甘南牦牛中,HaeⅢA基因型(175 bp/85 bp/35 bp)为优势基因型,基因型频率为0.306。3个群体的多态信息含量分别为0.739,0.754,0.743,均达到了高度多态(PIC>0.50)。说明牦牛DRB3.2基因具有高度多态性,在研究牦牛抗病育种和提高牦牛生产性能方面具有独特的效力和广泛的应用前景。     

成都麻羊和波尔山羊生长激素基因HaeⅢ多态性的比较研究

利用PCR-RFLP技术检测了成都麻羊和波尔山羊生长激素（Growth Hormone，GH）基因的HaeⅢ酶切多态性。结果表明：成都麻羊和波尔山羊2个山羊群体中均存在GH基因HBeⅢ酶切位点的多态性，且均有A、B两个等位基因。成都麻羊群体等位基因A和B的基因频率分别为0．6216和0．3784；波尔山羊群体中等位基因A和B的基因频率分别为0．5345和0．4655。GH基因HaeⅢ酶切住点的基因型分布在两个山羊群体中均极显著偏离Hardy-Weinberg平衡定理（P〈0．01）。但两个群体间不存在显著差异（P〉0．05）。     

陕西地区荷斯坦牛Mx1基因外显子多态性分析

牛Mx1基因位于1号染色体上,含有14个外显子,编码648个氨基酸。研究以346头陕西地区荷斯坦牛为研究对象,根据GenBank所提供的牛Mx1基因序列,设计15对引物,采用混合DNA池扩增测序和PCR-RFLP的方法对牛Mx1基因的14个外显子进行多态性分析。结果表明,4个SNPs,分别位于外显子3（g.843330T〉C,g.843411A〉G）、外显子4（g.844048C〉T）和外显子7（g.851325T〉C）。其中,外显子4（g.844048C〉T）为同义突变;外显子3（g.843330T〉C,g.843411A〉G）两个位点的突变分别导致了异亮氨酸（Ile）向苏氨酸（Thr）、谷氨酸（Glu）向精氨酸（Arg）的突变;外显子7（g.851325T〉C）位点的突变导致亮氨酸（Leu）变为脯氨酸（Pro）。4个SNPs位点都存在三种基因型,但纯合子的突变频率较低。4个SNPs位点共构建了11种单倍型,其中以H6为最主要的单倍型。结果显示,陕西地区荷斯坦牛Mx1基因存在较丰富的多态性,这将为Mx1基因结构和功能的研究以及将来在育种实践中的应用提供基础资料。     

The effect of lighting conditions on the rhythmicity of growth hormone secretion in Holstein steers

Growth hormone (GH) secretion regularity and the effects of lighting condition and GH‐releasing hormone (GHRH) on GH release were determined in steers. First, steers were kept under 12:12 L : D conditions (light: 06.00–18.00 hours). The animals were then subjected to a 1‐h advancement in lighting on/off conditions (05.00 and 17.00 hours, respectively). Blood was sampled for 24 h at 1‐h interval on the seventh day of each condition. Second, GHRH was injected intravenously (IV) at 12.00 and 00.00 hours under 12:12 L : D and blood was sampled at 15‐min interval for 4‐h (1 h before and 3 h after the injection). Plasma GH concentrations were measured by a radioimmunoassay. Periodicity of GH secretory profile was calculated by power spectrum analysis using the maximum entropy method. Plasma GH concentrations showed a characteristic pattern consisting of four distinct peaks. Mean periodicity of GH secretory profile was 5.7 h, and it was not altered by any change in lighting conditions. IV injection of GHRH increased GH secretion during the day and night. The increase in GH secretory volume after GHRH injection during the night was equal to that during the day. The present results suggest that GH secreted from the anterior pituitary have regularity in steers.     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

Effect of cellooligosaccharide or synbiotic feeding on growth performance,fecal condition and hormone concentrations in Holstein calves

We investigated the effect of cellooligosaccharide (CE) or a combination of dextran and Lactobacillus casei ssp. casei strain JCM1134^T (synbiotic; SB) feeding on growth performance, fecal condition and hormone concentrations in Holstein calves. Fifty‐two female Holstein calves were randomly assigned to three treatment groups: CE feeding group (n = 16), SB feeding group (n = 18), and control group (n = 18). Body weight at 90 days of age, as well as daily body weight gain (DG) and feed efficiency after weaning to 90 days of age were greater (P < 0.05) in the CE feeding group than in the control group. The total fecal score tended to be lower (P < 0.1) in the SB feeding group than in the control group. Plasma insulin concentration was higher (P < 0.05) in the CE feeding group than in the control group at 90 days of age. Our results indicate that CE feeding improved DG and feed efficiency in calves. On the other hand, there was less effect on growth performance and fecal Escherichia coli counts in calves fed SB.     

The effects of l‐DOPA and sulpiride on growth hormone secretion at different injection times in Holstein steers

The effects of l ‐DOPA, a precursor of dopamine (DA), and sulpiride, a D₂‐type DA receptor blocker, on growth hormone (GH) and prolactin (PRL) secretion were investigated in steers. Eight Holstein steers (212.8 ± 7.8 kg body weight) were used. Lighting conditions were 12:12 L:D (lights on: 06.00–18.00 hours). Blood samplings were performed during the daytime (11.00–15.00 hours) and nighttime (23.00–03.00 hours). Intravenous injections of drugs or saline were performed at 12.00 hour for the daytime and 00.00 hour for the nighttime, respectively. Plasma GH and PRL concentrations were determined by radioimmunoassay. l ‐DOPA did not alter the GH secretion when it was injected at 12.00 hour (spontaneous GH level at its peak). On the other hand, l ‐DOPA increased GH secretion at 00.00 hour (GH level at its trough). Injection of sulpiride suppressed GH secretion at 12.00 hour but did not affect GH levels at 00.00 hour. l ‐DOPA inhibited and sulpiride stimulated PRL release during both periods. These results suggest that dopaminergic neurons have stimulatory action on GH secretion and inhibitory action on PRL secretion in cattle. In addition, injection time should be considered to evaluate the exact effects on GH secretion due to its ultradian rhythm of GH secretion in cattle.     

Polymorphisms in growth hormone gene and their associations with calf weight in Japanese Black cattle

The objectives of this study were to detect effective genetic polymorphisms of bovine growth hormone (bGH) gene associated with calf weight in Japanese Black cattle. Fifty‐eight sires and 47 breeding cows were used to detect the polymorphisms in exons by single‐strand conformation polymorphism (SSCP). Four homozygous and six heterozygous SSCP genotypes were identified in exon 5. Although each single nucleotide polymorphism (SNP) had been reported, these genotypes were caused by three SNPs at the nucleotide positions 2141, 2277 and 2291. Four haplotypes C‐C‐A, G‐C‐A, C‐C‐C and G‐T‐A were newly identified. It was suggested that other haplotypes not detected in this study may not exist, considering the allele frequencies reported in Bos taurus and Bos indicus, and the migrating process of native Japanese cattle. Thereafter, we examined associations between the detected polymorphic sites in exon 5 by PCR – restriction fragment length polymorphism and calf weight using 53 breeding dams and 135 calves. The birth weights of calves with haplotype G‐C‐A are significantly lighter and calves' weights produced by cows with such haplotype are also lighter at 30 days old, using regression analysis. Although further research is necessary, these results may serve as a useful criterion to select breeding stocks, especially in maternal abilities.     

金华猪生长激素基因多态性分析及生长曲线比较

采用PCR-RFLP法测定了金华猪生长激素基因Apa -Ⅰ及Msp Ⅰ酶切多态性.并分析了各基因型与生长速度的相关性,进而对金华猪的生长曲线进行了拟合.结果表明,金华猪生长激素基因经Apa Ⅰ酶切后未出现多态,经Msp Ⅰ酶切后出现了AB(274、238、137 bp)和BB(238、137 bp)2种基因型,且不同基因型间的个体初生体质量、1月龄体质量、2月龄体质量、3月龄体质量、5月龄体质量、6月龄体质量及0～6月龄间平均日增体质量的差异不显著(P>0.05),而BB基因型4月龄体质量差异高于AB基因型(P<0.05).生长曲线拟合结果表明,Bertalanffy模型效果最佳.AB与BB基因型2种生长曲线进行比较,显示出BB基因型具有早期生长速度较快、生长拐点早、拐点体质量小的特点.     

肠出血性大肠杆菌O157毒力及黏附相关基因的PCR检测和PCR—RFLP分析

为了了解大肠杆菌O157分离菌株携带毒力及黏附相关基因的情况及菌株的多态性.用聚合酶链式反应扩增stx1、stx2、eaeA、ehxA、EspA和Tccp基因.选用限制性内切酶HincⅡ和EcoRⅡ对分离的O157:H7菌株eaeA基因进行酶切,选用限制性内切酶HaeⅢ、HinfⅠ、EcoRⅡ和RsaⅠ对分离的O157:H7菌株chxA基因进行酶切,最后对这两个基因进行PCR-RFLP分析.结果,所有O157:H7菌株扩增出eaeA、ehxA、EspA和Tccp基因,但没有扩增出stx1和stx2基因;4株O157:H?菌株,均没有扩增出stx2、eaeA、ehxA、EspA和Tccp基因,且只有1株扩增出stx1基因.所有分离菌株的eaeA基因和ehxA基因选用限制性内切酶酶切之后所得酶切片段数目和大小与标准菌株的eaeA基因和ehxA基因选用限制性内切酶酶切之后所得酶切片段数目和大小相同.结果表明,由于所有菌株缺失stx2基因,其致病力相对较低,且在基因水平较为保守,多态性较为单一.