耐氟喹诺酮类药物大肠埃希氏菌基因突变的MAMA-PCR检测

采用MAMA-PCR法对72株已知突变菌株的突变情况进行了检测。结果，有2种gyrA83位突变：TCG→TTG和TCG缺失；4种gyrA87位突变：GAC→AAC、GAC→GGC、GAC→TAC和GAC→CAC；1种parC80位突变：AGC→ATC以及4种parC84位突变：GAA→AAA、GAA→GCA、GAA→GGA和GAA→GTA。采用同样的方法，对38株未知突变菌株的检测结果显示，gyrA83、gyrA87、parC 80和parC84位的突变检出的正确率分别为97．4％、94．7％、81．6％和94．7％。结果表明，MAMA-PCR有望成为监测耐氟喹诺酮类药物大肠埃希氏菌的分子生物学常规方法。     

Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats

The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis.     

日粮蛋氨酸水平对大骨鸡肌肉SLC12A5、 Musclin mRNA表达影响的研究

试验旨在研究日粮蛋氨酸水平大骨鸡腿肌和胸肌SLC12A5、Musclin mRNA表达量的影响.选择240只16周龄公鸡,随机分为3组,每组10个重复,每个重复8只.H组、M组和L组日粮中蛋氨酸(Met)水平分别为0.42％、0.34％、0.25％,其中M组为对照组.试验期8w.结果 显示,H组各周龄公鸡血清Met含量...     

季节放牧下青藏高原高寒草甸牧草生物量空间分布特征

高寒草甸是青藏高原的主要草地类型之一,对维持国家生态屏障和当地人民生活具有极为重要的意义.本研究以甘肃玛曲高寒草甸4个季节性牦牛牧场为研究对象,探讨了季节性放牧对牧草生物量空间分布的作用.结果表明,不放牧样地和全年连续放牧样地植物群落地上生物量的变异系数高于其他季节牧场,分别为29.06%和29.02%.不放牧样地的根...     

云南乳饼加工技术及品质影响因素

云南乳饼作为具有少数民族特色的传统发酵乳制品,其品质优良且工艺简单,极具市场发展前景,但目前仍处于传统手工作坊式阶段,急需实现乳饼的标准化生产。本文首先总结分析了目前乳饼的生产工艺,对其中关键工艺进行阐述,为工艺产业化提供参考;其次综述了影响乳饼品质的因素,包括鲜乳稀释比、凝乳温度和pH、凝乳剂、挤压强度和时间以及外源添加剂等,深入介绍这些因素对工艺及产品质量的影响,旨在为开展乳饼的新工艺和新产品研发提供理论依据,以期将富含民族特色的乳饼推向市场销售。     

播种期对当归种苗生理指标及质量的影响

开展当归不同播种期育苗试验,探究播种期所对应气候因子对当归种苗生理指标及质量的影响,寻找当归育苗的最佳气候条件。试验结果显示,各气候因子与种苗单根重、根粗、产量、一级苗比例、成活率和早期抽薹率之间均存在显著或极显著正相关关系,平均地温、平均气温、≥10 ℃有效积温及降水量与丙二醛(MDA)含量呈显著负相关关系,与过氧化氢酶(CAT)、过氧化物酶(POD)活性呈显著正相关关系。6 月 20 日(第 2 播种期)播种的种苗产量分别比 6 月 10 日、6 月 30 日播种的种苗产量高 11. 96%、65. 63%,MDA 含量分别低 2. 02% 和 29. 44%,超氧化物歧化酶(SOD)、过氧化物酶 (POD)活性较高,药材产量达 1 691. 2 kg/hm² 。综合种苗生理指标及产量,6 月 20 日(第 2 播种期)播种的种苗抗逆性强,早期抽薹率低,药材产量高,可作为适宜播种期,其对应的气象因子可作为适宜气候条件。     

抗丝状支原体丝状亚种单克隆抗体的制备及初步应用

旨在制备抗丝状支原体丝状亚种(Mmm)的特异性单克隆抗体(MAb),为牛传染性胸膜肺炎(CBPP)病原诊断的免疫学方法提供特异性抗体。本研究利用生物信息学技术分析了Mmm国内分离株Ben-1不同传代株的全基因组序列,选取M0071蛋白作为研究对象。将原核表达的可溶性重组蛋白M0071(rM0071)作为免疫原免疫BALB/c小鼠,通过有限稀释法和间接ELISA方法筛选得到能稳定分泌抗rM0071蛋白的单克隆抗体的杂交瘤细胞株。进一步制备单抗腹水并纯化,利用Western blot方法对该单抗进行特异性鉴定,同时测定其抗体效价和抗体亚类。随后利用间接免疫荧光试验(IFA)评价该单抗对细胞感染Mmm的检测能力。结果表明成功获得1株单克隆细胞株3C4A1,将其分泌抗体命名为MAb 3C4A1。特异性结果表明,MAb 3C4A1能与Mmm的分离株和标准株发生特异性反应,而不与山羊支原体山羊肺炎亚种、丝状支原体山羊亚种、牛鼻支原体、无乳支原体、牛支原体、leachii支原体和牛A型巴氏杆菌等发生反应。抗体亚类鉴定MAb 3C4A1属于IgG1亚类、轻链为κ链。经间接ELISA测定其抗体效价为1∶256 000。IFA试验结果表明,MAb 3C4A1仅与感染EBL细胞的Mmm发生绿色荧光反应,而与牛鼻支原体、无乳支原体、牛支原体感染的细胞不发生荧光反应,特异性良好。本研究制备的MAb 3C4A1具有良好的特异性和免疫反应性,可作为CBPP病原免疫学诊断的工具,为进一步研制CBPP病原鉴别诊断试剂盒提供了基础材料。     

Building phenotype networks to improve QTL detection: a comparative analysis of fatty acid and fat traits in pigs

Models in QTL mapping can be improved by considering all potential variables, i.e. we can use remaining traits other than the trait under study as potential predictors. QTL mapping is often conducted by correcting for a few fixed effects or covariates (e.g. sex, age), although many traits with potential causal relationships between them are recorded. In this work, we evaluate by simulation several procedures to identify optimum models in QTL scans: forward selection, undirected dependency graph and QTL-directed dependency graph (QDG). The latter, QDG, performed better in terms of power and false discovery rate and was applied to fatty acid (FA) composition and fat deposition traits in two pig F2 crosses from China and Spain. Compared with the typical QTL mapping, QDG approach revealed several new QTL. To the contrary, several FA QTL on chromosome 4 (e.g. Palmitic, C16:0; Stearic, C18:0) detected by typical mapping vanished after adjusting for phenotypic covariates in QDG mapping. This suggests that the QTL detected in typical mapping could be indirect. When a QTL is supported by both approaches, there is an increased confidence that the QTL have a primary effect on the corresponding trait. An example is a QTL for C16:1 on chromosome 8. In conclusion, mapping QTL based on causal phenotypic networks can increase power and help to make more biologically sound hypothesis on the genetic architecture of complex traits.     

旋壳转速对腔内液体流动特性的效应

为研究旋壳转速对腔内液体流动特性的影响,以试验旋喷泵为研究对象,在高度验证叶轮与旋壳同步旋转试验与模拟结果准确性的基础上,对叶轮转速相同、旋壳转速不同的5个模型采用RNG k-ε湍流模型进行数值计算,分析腔内液体流动特性的变化情况,研究泵的性能.结果表明:旋壳转速增大,液体圆周速度和旋转系数均增大,圆周速度曲线沿径向逐渐形成同心圆,腔内液体做非刚性旋转.腔内液体径向压力梯度增大,压力低于624 kPa时,旋壳转速越高,压力越小;压力高于624 kPa时,旋壳转速越高,压力越大.集流管迎流区涡分布在进口附近,尾迹区涡集中在扩散段结尾处,整体呈增大趋势.旋壳转速增大,泵的扬程升高,但效率降低,通过改变集流管进口直径发现集流管并非效率降低的主要原因,而是由圆盘摩擦损失的增大导致的,圆盘摩擦损失随旋壳转速增加呈3次幂函数式增大,文中最优进口直径为13 mm.     

中国主栽水稻品种微卫星标记数据库的初步构建

以前期筛选的微卫星标记为基础,并在4个染色体上增加新标记,进一步检测了63个主栽常规稻品种和杂交稻亲本,推荐了24个微卫星标记（每条染色体2个）作为水稻品种鉴别的标记;除了63个常规稻品种和杂交稻亲本外,这24个标记还应用于检测41个主栽杂交稻组合,并确定了各个标记在试验材料中检测到的等位基因类型。通过杂交稻组合与其亲本的标记匹配性检验,确认了杂交稻母本的高真实性,提高了恢复系数据的可靠性,剔除了1个假杂交稻,建立了103个水稻材料×24个标记的主栽水稻品种微卫星标记数据库,并分析了数据库中各个标记、各组材料的多态性表现。此外,还讨论了材料真实性和典型性对数据库构建的影响,提出了水稻品种间差异判别标准的建议及其应用风险和解决途径。