中国林蛙抗菌肽Temporin-1 CEa基因的真核表达载体构建

为了建立大量获取中国林蛙抗菌肽的方法,根据GenBank中的中国林蛙皮肤抗菌肽Temporin-1CEa基因的mRNA序列(EU624139)设计一对特异性引物,以提取的中国林蛙皮肤总RNA反转录出的cDNA为模板,将扩增的编码序列与pEASY-T3克隆载体连接获得Tem-T3;利用酶切、连接等分子生物学手段,将GFP基因连入Tem-T3克隆载体,再经酶切获得Tem-GFP片段,并插入真核表达载体pcDNA3.1,最终得到Tem-GFP-pcD-NA3.1重组质粒;利用脂质体转染法将该质粒转入绵羊成纤维细胞,48 h后可在荧光倒置显微镜下观察到GFP的绿色荧光表达;qPCR数据分析显示,与对照组相比,转染Tem-GFP-pcDNA3.1的绵羊成纤维细胞中融合蛋白Tem-GFP的表达量可提高约300倍。本研究为构建Temporin-1CEa基因山羊乳腺特异表达载体提供依据。     

晋中绵羊耳成纤维细胞系建立

以晋中绵羊耳缘组织为材料,采用组织块贴壁培养法,通过原代培养和传代培养对细胞进行了细胞形态学观察、细胞计数和生长曲线的绘制,探讨了细胞体外培养模式.结果:细胞生长总体趋势呈“S”型;建立了晋中绵羊耳成纤维细胞系,细胞系的建立,使晋中绵羊的重要种质资源在细胞水平上保存下来.     

DNA polymorphism of 5' flanking region of prolactin gene and its association with litter size in sheep

A single nucleotide polymorphism of 5' flanking region of the prolactin gene was investigated in both high prolificacy breeds (Small Tail Han and Hu sheep) and low prolificacy breeds (Dorset and Suffolk sheep) using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP). The results indicated that two genotypes (AA and AB) were detected in Small Tail Han sheep (n   =   239), only one genotype (AA) was detected in Hu (n   =   40), Dorset (n   =   50) and Suffolk sheep (n   =   39). The mutant homozygous genotype (BB) was not detected in four sheep breeds. In Small Tail Han sheep (n   =   239), the frequency of genotypes AA and AB was 0.91 and 0.09, the frequency of the A and B alleles was 0.95 and 0.05, respectively. The fitness tests showed that the Small Tail Han sheep population was in Hardy–Weinberg equilibrium. Sequencing revealed a mutation (G→T) at the position 63 bp of the 5' flanking region of prolactin gene in AB genotype compared with AA genotype in Small Tail Han sheep. The Small Tail Han ewes with AB genotype had 0.83 (p < 0.05) lambs more than those with AA genotype. These results preliminarily showed that the prolactin locus is either a major gene that influences the high prolificacy in Small Tail Han sheep or is in close linkage with such a gene.     

Estimation of (co)variance components and genetic parameters of greasy fleece weights in Muzaffarnagari sheep

Variance components and genetic parameters for greasy fleece weights of Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Mathura, India, over a period of 29 years (1976 to 2004) were estimated by restricted maximum likelihood (REML), fitting six animal models including various combinations of maternal effects. Data on body weights at 6 (W6) and 12 months (W12) of age were also included in the study. Records of 2807 lambs descended from 160 rams and 1202 ewes were used for the study. Direct heritability estimates for fleece weight at 6 (FW6) and 12 months of age (FW12), and total fleece weights up to 1 year of age (TFW) were 0.14, 0.16 and 0.25, respectively. Maternal genetic and permanent environmental effects did not significantly influence any of the traits under study. Genetic correlations among fleece weights and body weights were obtained from multivariate analyses. Direct genetic correlations of FW6 with W6 and W12 were relatively large, ranging from 0.61 to 0.67, but only moderate genetic correlations existed between FW12 and W6 (0.39) and between FW12 and W12 (0.49). The genetic correlation between FW6 and FW12 was very high (0.95), but the corresponding phenotypic correlation was much lower (0.28). Heritability estimates for all traits were at least 0.15, indicating that there is potential for their improvement by selection. The moderate to high positive genetic correlations between fleece weights and body weights at 6 and 12 months of age suggest that some of the genetic factors that influence animal growth also influence wool growth. Thus selection to improve the body weights or fleece weights at 6 months of age will also result in genetic improvement of fleece weights at subsequent stages of growth.     

Calculating economic values for growth and functional traits in non‐dairy sheep

A bio‐economic model including a computer program was developed where a large number of parameters can be modified to describe a great variety of production systems. Economic values for up to 27 traits can be calculated. Seasonal feeding is considered and protein requirement in addition to net energy may be taken into account when calculating feed requirement. Furthermore, the model allows for the evaluation of individual parts of the growth curve. The model was applied to the Czech Suffolk breed used in pure‐breeding. An extensive pasture system with spring lambing and sale of surplus progeny for slaughter after weaning or after a finishing period was assumed. The economic values (in Euro cent per ewe, per year and per genetic standard deviation of the trait) were as follows: 9.9 for birth weight, ?72.6 for mature weight, 142, 32.1 and 16.9 for growth rates till weaning, in rearing of breeding animals and in fattening, respectively, 19.9 and 101 for conception rate of female lambs and ewes, respectively, 690 for litter size per ewe lambing, 172 for productive lifetime of ewes, 402 and 503 for survival rate of lambs at lambing and till weaning, respectively, and 16.5 for fleece weight.     

Direct and maternal (co)variance components and heritability estimates for body weights in Chokla sheep

Estimates of (co)variance components were obtained for weights at birth, weaning and 6, 9 and 12 months of age in Chokla sheep maintained at the Central Sheep and Wool Research Institute, Avikanagar, Rajasthan, India, over a period of 21 years (1980–2000). Records of 2030 lambs descended from 150 rams and 616 ewes were used in the study. Analyses were carried out by restricted maximum likelihood (REML) fitting an animal model and ignoring or including maternal genetic or permanent environmental effects. Six different animal models were fitted for all traits. The best model was chosen after testing the improvement of the log-likelihood values. Direct heritability estimates were inflated substantially for all traits when maternal effects were ignored. Heritability estimates for weight at birth, weaning and 6, 9 and 12 months of age were 0.20, 0.18, 0.16, 0.22 and 0.23, respectively in the best models. Additive maternal and maternal permanent environmental effects were both significant at birth, accounting for 9% and 12% of phenotypic variance, respectively, but the source of maternal effects (additive versus permanent environmental) at later ages could not be clearly identified. The estimated repeatabilities across years of ewe effects on lamb body weights were 0.26, 0.14, 0.12, 0.13, and 0.15 at birth, weaning, 6, 9 and 12 months of age, respectively. These results indicate that modest rates of genetic progress are possible for all weights.     

藏系绵羊钼诱导铜缺乏的研究

自2000年以来,青海省乌兰地区藏绵羊出现以异食、消瘦和贫血为特征的疾病。经分析该地区土壤、牧草和动物的矿物质元素含量,发现在患病区,土壤钼含量为（4.63±1.51）μg/g,牧草为（4.89±1.79）μg/g,极显著高于正常地区。患病动物血液平均铜含量为（0.23±0.07）μg/g,被毛铜平均含量为（3.27±0.71）μg/g,而正常区的动物为（0.97±0.23）μg/g和（6.78±1.57）μg/g,差异极显著;患病动物血液铜最高值为0.53μg/g,血液铜最低值为0.031μg/g。而正常地区藏绵羊血液铜含量最高值为1.33μg/g,血液铜最低值为0.83μg/g。同时患病动物血清铜蓝蛋白显著低于正常动物,并且有小红细胞贫血。结果表明,藏绵羊以异食、消瘦和贫血为特征的营养代谢疾病为钼诱导的次级铜缺乏。     

放牧绵羊微量元素摄入量与季节变化的关系研究

采用体质量-口罩法,分别在2、4、6、8、10、12月对天山北坡中段草地放牧的新疆细毛羊采食量进行测定,通过样方对牧草微量元素含量进行测定,计算放牧绵羊微量元素摄入量,另外不同季节绵羊发生周期性生理变化,对微量元素的需要量也不同,把不同季节放牧绵羊微量元素摄入量和需要量进行比较。研究结果得出不同季节绵羊从每天的采食中所获得的铁、锰、钴等微量元素能满足其需要。草地牧草中严重缺锌,母羊除在空怀期外其余时间锌缺乏量为16.0%82.2%,特别是在母羊泌乳前期只能从牧草中获得需要量17.8%的锌。绵羊怀孕后期和泌乳前期缺铜55%和59%,常年缺硒6%52%。     

Lythrum hyssopifolia (lesser loosestrife) poisoning of sheep in Victoria

Objective To document an ovine disease attributed to the consumption of Lythrum hyssopifolia (lesser loosestrife). Procedures Historical and histological review of field and experimental cases. Results 1–20% mortality occurred in sheep flocks grazing paddocks where L. hyssopifolia was the predominant green vegetation. Well‐documented disease outbreaks occurred in summer on nine farms across Victoria between 1974 and 2002. Liver damage occurred in all nine outbreaks, with kidney damage in at least eight. Hepatocyte necrosis was usually zonal to midzonal (zone 2) in the liver samples from four farms and periacinar (zone 3) in those from three farms, but some livers showed only single‐cell necrosis. Multinucleate hepatocytes near necrotic areas were a feature in six cases. Proximal tubular epithelium appeared to be the primary renal target and brown granules were often present in renal tubules. Biochemical and histological evidence of liver and kidney damage was obtained from two sheep experimentally pen‐fed harvested L. hyssopifolia. Conclusion Chemicals in L. hyssopifolia are toxic to ovine hepatocytes and renal tubular epithelial cells.     

西藏农区牛、绵羊体重与年龄拟合曲线回归分析

对西藏农区的母黄牛、黑白花母牛、西门塔尔母牛、茨盖羊、藏系母绵羊和藏系公绵羊进行体尺测定,以年龄为自变量,体重为因变量进行曲线拟合。拟合曲线的相关系数分别为0.83,0.72,0.78,0.68,0.67和0.74。对实测值和估测值之间进行F显著性检验,均差异不显著（P>0.05）,回归模型能够真实地反映西藏农区家畜的生长特点。