利用SSR、ISSR和RAPD技术构建苜蓿基因组DNA指纹图谱

在建立可靠的苜蓿基因组DNA提取分离和PCR扩增技术体系的基础上,筛选具有稳定多态性位点的RAPD、SSR和ISSR引物,建立苜蓿基因组DNA的SSR、ISSR和RAPD分子标记技术体系,并利用分子标记检测苜蓿品种(系)的DNA分子标记多态性,构建苜蓿品种的DNA指纹图谱.筛选出36个RAPD引物,8个SSR引物和12个ISSR随机引物,通过PCR扩增在55个国内外苜蓿品种(系)中获得了182个RAPD多态性位点和丰富的SSR和ISSR多态性位点,构建了55个苜蓿品种(系)的SSR、ISSR和RAPD指纹图谱.结果表明,不同苜蓿品种(系)的SSR多态性有较大差异;苜蓿品种ISSR指纹图谱多态性丰富,稳定性比RAPD强;可以通过2～3个引物鉴别我国主要苜蓿品种和引进品种,SSR和ISSR指纹图谱可以更好地用于苜蓿品种鉴定和遗传多样性分析.     

ISSR分子标记及其在动物遗传育种中的应用

近年来，分子遗传标记的研究与应用得到了迅速的发展。20世纪80年代中期以来，随着PCR技术的出现，基于PCR技术的分子标记，如随机括增多态性(randomam plified polymorphic DNA，RAPD)、小卫星(minisatellite)或称DNA指纹图谱(DNA fingerprinting，DFP)、微卫星(microsatellite)或称简单序列重复(simple sequence repeat，SSR)、括增片段长度多态性(amplified fragment lengt hpolymorphism，AFLP)、单核     

分子标记与地方猪品种的遗传多样性研究

分子标记已成为目前家畜遗传多样性研究的主要工具。本文简要综述了多种分子标记(RFLPs，nat-DNA，RAPD，DNA指纹图谱，微卫星DNA，AFLP，SNPs)的检测方法及其在地方猪品种遗传变异和遗传关系检测分析中的应用，为类似研究提供参考材料。     

用寡核苷酸探针对近交系小鼠DNA指纹图谱的分析

应用DNA指纹技术，用非放射性标记的寡核苷酸探针(GGAT)。对3个清洁级近交小鼠品系C57、DBA／2、BALB／c进行检测，分析其DNA指纹图谱。结果显示，不同品系近交系小鼠的DNA指纹图谱有明显差异，而品系内小鼠之间的DNA指纹图谱基本一致。证明DNA指纹技术能够较好地检测出近交系小鼠的基因纯合性，反映其遗传质量，可以应用于对近交系小鼠的遗传质量监测。     

苜蓿DUS测试标准品种SSR分子标记指纹图谱的构建

苜蓿在育种过程中,由于其异花授粉特性和骨干亲本的集中使用,导致品种间的遗传差异日益缩小,品种之间通过形态学鉴定愈加困难。通过构建苜蓿DUS测试标准品种DNA指纹图谱数据库,可以实现苜蓿品种的快速、准确鉴定。本研究利用筛选得到的10个SSR标记对33个苜蓿DUS测试标准品种进行指纹图谱构建和遗传多样性分析。结果表明,10对SSR引物共扩增出69个等位基因,平均每个标记可产生6.9个等位基因;多态性比率(PPB)从25%到90%不等,平均值为56.7%;各SSR标记的多态信息含量(PIC)范围为0.56~0.87,平均值为0.75。利用不同引物间的组合能有效区分33个苜蓿 DUS标准品种,并为每份标准品种建立了唯一标识的指纹代码,为苜蓿品种的审定和保护以及遗传背景分析提供了技术依据。     

柱花草DUS测试标准品种DNA指纹图谱构建

为建立柱花草(Stylosanthes spp.)DUS测试标准品种DNA指纹图谱数据库,实现柱花草品种的快速、准确鉴定,利用筛选到的25个SSR标记对15份柱花草DUS测试标准品种进行了遗传多样性分析及DNA指纹图谱构建.结果表明,25个SSR标记共产生132个等位基因,每个标记可产生3～9个等位基因,平均5.280个;各SSR标记的多态性信息含量为0.393～0.847,平均为0.638.利用这25个SSR标记构建的指纹图谱能有效区分15份柱花草DUS标准品种,并为每份标准品种建立了QR二维码图谱,为SSR标记技术在柱花草DUS测试中的应用提供依据.     

西门塔尔牛、和牛与荷斯坦牛杂种优势预测及实际杂交效果分析

旨在利用覆盖全基因组和性状的特异性SNPs标记预测和牛、西门塔尔牛与荷斯坦牛杂种优势,为牛杂种优势利用和选种选配提供参考依据。本研究分别利用牛Illumina Bovine HD 770 K和GGP Bovine 100 K芯片对464头和牛、1 222头西门塔尔牛和43头荷斯坦牛3个亲本群体进行基因型分型,并通过牛QTLs数据库筛选与目的性状对应的QTLs,对比牛参考基因组映射得到与初生重、周岁重、胴体重性状相关的特异性SNPs;然后构建覆盖全基因组和性状特异性SNPs两种标记状态同源矩阵,通过计算杂交组合亲本间的遗传距离来预测品种间杂种优势,并利用配合力分析验证较优组合的实际杂交效果。结果表明,基于全基因组和性状特异性SNPs计算的各杂交组合遗传距离差异不显著。在全基因组水平上,西门塔尔牛♂×荷斯坦牛♀（S×H）与和牛♂×荷斯坦牛♀（W×H）亲本间杂交组合遗传距离分别为0.346 1和0.338 9;在初生重、周岁重和胴体重性状上,S×H亲本间遗传距离分别为0.343 1、0.348 7和0.336 7,而W×H遗传距离分别为0.337 6、0.340 7和0.329 2;两种SNPs标记计算的遗传距离均为S×H较大,W×H次之。因此,在初生重、周岁重、胴体重性状上,S×H为较优杂交组合。通过分析德系西门塔尔牛♂×荷斯坦牛♀实际杂交群体的配合力,发现10个父系在初生重性状上一般配合力和特殊配合力均为正效应,最高效应值分别达到3.760 9和8.931 2。西门塔尔牛与荷斯坦牛杂交可在初生重、周岁重和胴体重获得较高的杂种优势。     

南阳牛分子标记研究进展

分子标记技术的开发利用在家畜育种中起着越来越重要的作用.南阳黄牛是中国地方良种黄牛之一.近些年来,已在DNA水平上对南阳牛进行了大量的研究,涉及到南阳牛的分子群体遗传特征及起源进化、经济性状分子遗传标记及相关功能基因多态性研究等.本文对南阳牛分子标记研究方面的成就作了综述,以便促进南阳牛的保护、利用和发展.使传统育种和分子标记辅助选择相结合,推动了南阳牛的育种进程.     

湖北省现行家蚕品种原原种AFLP指纹图谱的构建

利用AFLP分子标记技术,构建了湖北省10个优质强健原原种的AFLP指纹图谱,分析了其遗传相似系数及亲缘关系。利用4对AFLP引物,共扩增了584条带,其中有多态性带354条,多态性比例为60.62%,构建了其AFLP指纹图谱,可以在分子水平上对这10个家蚕原原种进行鉴定和检测。     

延边牛Y染色体遗传多样性与父系起源研究

[目的]为了研究延边牛Y染色体遗传多样性以及父系起源。[方法]采用PCR扩增、直接测序和生物信息学方法,分析延边牛的2个Y-SNPs标记(UTY19和ZFY10)和2个Y-STRs标记(INRA189和BM861)的遗传多样性。[结果]Y-SNPs标记检测发现延边牛有Y1和Y2单倍型组,2个Y-STRs标记在延边牛品种检测发现只有INRA189表现多态性(98bp/102bp/104bp)。结合Y-SNPs与Y-STRs的分型结果,确定延边牛有3种Y染色体单倍型Y1-98-158、Y2-102-158和Y2-104-158,所占频率分别为0.031、0.938和0.031,其Y染色体单倍型多样度为0.1230±0.0777,表明延边牛父系遗传多样性很低,父系遗传稳定。[结论]延边牛为普通牛父系起源,其中Y2单倍型组占明显优势。     

Concentrations of strontium,barium, cadmium,copper, zinc,manganese, chromium,antimony, selenium,and lead in the liver and kidneys of dogs according to age,gender, and the occurrence of chronic kidney disease

This study was conducted to measure the concentrations of strontium (Sr), barium (Ba), cadmium (Cd), copper (Cu), zinc (Zn), manganese (Mn), chromium (Cr), antimony (Sb), selenium (Se), and lead (Pb) in canine liver, renal cortex, and renal medulla, and the association of these concentrations with age, gender, and occurrence of chronic kidney disease (CKD). Tissues from 50 dogs were analyzed using inductively coupled plasma mass spectrometry. Cu, Zn, and Mn levels were highest in the liver followed by the renal cortex and renal medulla. The highest Sr, Cd, and Se concentrations were measured in the renal cortex while lower levels were found in the renal medulla and liver. Female dogs had higher tissue concentrations of Sr (liver and renal medulla), Cd (liver), Zn (liver and renal cortex), Cr (liver, renal cortex, and renal medulla), and Pb (liver) than male animals. Except for Mn and Sb, age-dependent variations were observed for all element concentrations in the canine tissues. Hepatic Cd and Cr concentrations were higher in dogs with CKD. In conclusion, the present results provide new knowledge about the storage of specific elements in canine liver and kidneys, and can be considered important reference data for diagnostic methods and further investigations.     

Comparisons of Angus, Charolais, Galloway, Hereford, Longhorn, Nellore, Piedmontese, Salers, and Shorthorn breeds for weight, weight adjusted for condition score, height, and condition score of cows

Breed differences for weight (CW), height (CH), and condition score (CS) were estimated from records (n = 12,188) of 2- to 6-yr-old cows (n = 744) from Cycle IV of the U.S. Meat Animal Research Center's Germplasm Evaluation (GPE) Program. Cows were produced from mating Angus and Hereford dams to Angus, Hereford, Charolais, Shorthorn, Galloway, Longhorn, Nellore, Piedmontese, and Salers sires. Samples of Angus and Hereford sires were 1) reference sires born from 1962 through 1970 and 2) 1980s sires born in 1980 through 1987. The mixed model included cow age, season of measurement and their interactions, year of birth, pregnancy-lactation code (PL), and breedgroup as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as a linear covariate. The model for CH excluded PL. Random effects were additive genetic and permanent environmental effects associated with the cow. Differences among breed groups were significant (P < 0.05) for all traits and were maintained through maturity with few interchanges in ranking. The order of F1 cows for weight was as follows: Charolais (506 to 635 kg for different ages), Shorthorn and Salers, reciprocal Hereford-Angus (HA) with 1980s sires, Nellore, HA with reference sires, Galloway, Piedmontese, and Longhorn (412 to 525 kg for different ages). Order for height was as follows: Nellore (136 to 140 cm), Charolais, Shorthorn, Salers, HA with 1980s sires, Piedmontese, Longhorn, Galloway and HA with reference sires (126 to 128 cm). Hereford and Angus cows with reference sires were generally lighter than those with 1980s sires. In general, breed differences for height followed those for weight except that F1 Nellore cows were tallest, which may in part be due to Bos taurus-Bos indicus heterosis for size.     

Supplementation of broodmares with copper, zinc, iron, manganese, cobalt, iodine, and selenium

In experiment 1, 6 pregnant mares received a concentrate that contained a trace mineral premix that provided 14.3 mg Cu, 40 mg Zn, 28 mg Fe, 28 mg Mn, 0.08 mg Co, 0.16 mg I, and 0.16 mg Se/kg concentrate (group A). Seven mares received the same concentrate plus 502 mg Zn and 127 mg Cu once daily (group B). No differences (P > .05) in foal growth data, or Cu, Zn, and Fe concentrations of mare milk, mare serum, or foal serum were observed. In experiment 2, 6 pregnant mares received the same concentrate as group A (group C), and 8 mares received the same concentrate fortified with 4× the trace mineral premix (group D). Group C mares had higher serum Zn concentration at 1 day (P < 0.01) and 56 days (P < 0.04). Group C mares had higher milk Fe concentration at 28 days (P < .01), and group D mares had higher milk Cu concentration at 56 days (P < .01). Group C foals had higher serum Cu concentration at 14 days (P < .03). The results from this study provide no evidence to indicate that supplementing late gestating and lactating mares with higher dietary trace mineral levels than those recommended currently by NRC has any influence on foal growth and development, or on the Cu, Zn, and Fe concentrations of the mare milk, mare serum, or foal serum.     

电感耦合等离子-质谱法测定饲料中钠、镁、铬、锰、铁、铜、锌、砷、硒、镉和铅

应用电感耦合等离子-质谱技术(ICP-MS),建立饲料中钠、镁、铬、锰、铁、铜、锌、砷、硒、镉和铅等元素的测定方法。对饲料样品的前处理方法、仪器工作参数和11种元素标准曲线进行优化;并以加标回收、分析方法比对和重复测试说明方法的准确性和精密性。方法在0～1 000 ng/ml范围内线性良好,仪器检出限为0.557 7～5.072 ng/ml,具有良好的精密度,其回收率在88.1%～104.4%之间,相对标准偏差小于5.0%。同时与原子吸收和原子荧光方法进行比对,测定结果相近。所建立的方法简单、快速,可替代原子吸收和原子荧光方法测定饲料中的11种金属元素,为饲料的质量控制提供理想的元素分析方法。     

Granulocyte isolation from whole blood of goat, sheep, cattle, horse, dog, pig, and man

A simple two step procedure for the isolation of caprine, ovine, bovine, equine, canine, porcine and human peripheral blood granulocytes is described. After enrichment of granulocytes by centrifugation, contaminating erythrocytes are lysed hypotonically. Recovery, purity, and viability of the granulocyte suspensions are determined. FACScan analysis of the cell suspensions measuring cellular size by forward and sideward light scatter is compared with the corresponding analysis of whole blood leukocytes. Constituencies of the isolated cell suspensions and loss of granulocyte subpopulations through isolation procedure is discussed with regard to granulocyte function assays.     

The effects of antibiotic,probiotic, organic acid,vitamin C,and Echinacea purpurea extract on performance,carcass characteristics,blood chemistry,microbiota, and immunity of broiler chickens

The present context investigated the comparative study on the supplementation of antibiotic, probiotic, organic acid, vitamin C, and herbal extract after vaccination into drinking water and their effects on performance, carcass quality, blood biochemical parameters, immune system, and intestinal flora in broiler chicks for 42 days. A total of 420 one-day-old male broiler chicks (Ross 308) were randomly assigned into 7 treatments with 3 replicates (pens) per treatment and 20 male chicks for each replicate (pen). The experimental treatments consisted of drinking water (control, without additive); drinking water + antibiotic sulfamet; drinking water + C-Vet-50; drinking water + antibiotic sulfamet + C-Vet-50; drinking water + probiotic Primalac; drinking water + butyric acid; and drinking water + extract of Echinacea purpurea Moench (coneflower). There were no differences observed among the treatments for feed intake, but during the whole experimental period, the highest body weight gain was found in the chicks fed with drinking water + antibiotic sulfamet + 50 cc vitamin C (P < 0.05). There were no differences (P > 0.05) observed among the treatments for feed conversion ratio (P > 0.05). Moreover, there were no differences reported among treatments for carcass characteristics at the end of the experiment. Among the treatments, drinking water + 50 cc vitamin C, and drinking water + extract of E. purpurea reduced (P < 0.05) the levels of cholesterol, triglycerides, and low-density lipoproteins. Drinking water + 50 cc vitamin C, drinking water + Primalac, and drinking water + extract of E. purpurea increased (P < 0.05) the lymphocytes count and decreased the heterophils count and heterophil:lymphocyte ratio. The highest Escherichia coli count and lowest Lactobacillus count in ileal content of the broilers were observed in the control group (P < 0.05). The additives used in this study may be incorporated in the drinking water of broiler chickens as growth promoters and for improved performance. A further, wider supplementation study is required to understand the performance, immune system, variation in the intestinal microbial counts, and any other possible alteration in the intestinal biota of the broilers.     

Department of Clinical Veterinary Practices, School of Veterinary Medicine, University of Dublin, Trinity College, Ballsbridge, Dublin 4

Circular excised skin wounds in the thoracic and metatarsal regions of the dog were studied. A similar sequence of events took place in the two regions although differences did occur due to the different reactions of the tissues which surrounded the wounds. None of the wound cavities became filled with exudate during the early stages of healing. In the thoracic wounds the cavities were largely filled by the swelling and inward movement of adipose tissue. Epithelium first grew on the wound surface in the sector of the wound that was situated in the direction of hair flow. The average time to complete epithelization was similar in both sets of wounds. A zone of alopecia developed around the wounds.