牦牛源正呼肠孤病毒2型的检测和分离鉴定

旨在调查川西北牦牛哺乳动物正呼肠孤病毒（MRV）的感染情况并分离病毒。采用RT-PCR方法，对采自川西北15个牧场的72份牦牛腹泻粪便样本和其中5个牧场的15份腹泻牦牛血清样本进行MRV检测，阳性样本进一步用分型PCR确定其血清型。结果显示，粪便样本中MRV检出率为20.83%（15/72），血清2型的比例为60%（9/15）；血清样本中MRV检出率为40%（6/15），血清2型的比例为83.33%（5/6）；未检测到其他血清型。成功地从腹泻粪便中分离到1株MRV血清2型毒株（TCID₅₀为4×10^-8.56·mL^-1），并获得长度为23 587 bp的分离株全基因组，该分离株与中国猪源毒株的遗传关系最近；与GenBank中所有的MRV S1基因相比，该分离株有4个独特的氨基酸突变。本研究从牦牛中检测到MRV，并分离到1株牛源MRV血清2型毒株，为进一步研究MRV血清2型生物学特性奠定了基础。     

转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。     

饲料中无机元素分析方法研究进展

饲料中所含的无机元素对动物生长具有重要意义。在饲料中过量添加无机元素会导致环境污染问题,也会通过食物链最终危害消费者的身体健康,因此,对饲料产品中的无机元素进行检测和动态监控尤为重要。综述了近年来饲料中无机元素含量检测主要应用的前处理方法及其含量检测技术,分析了不同前处理方法和含量检测技术的优点和不足,以期为饲料中无机元素分析方法的科学、合理应用提供参考。     

Detect unique molecular structure associated with physiochemical properties in CDC varieties of oat grain with unique nutrient traits [Feed Type vs. Milling Type] in comparison with barley grain using advanced molecular spectroscopy as a non-destructive biological tool

The objective of this study was to determinate grain unique protein inherent molecular structure that are related physiochemical and nutrient profiles in CDC developed oat varieties [CDC Nasser (Feed Type) and CDC Seabiscuit (Milling Type)] grown in cool climate condition in western Canada in comparison with conventional barley variety of CDC Meredith as a control using advanced molecular spectroscopy. Multivariate analyses, including an agglomerative hierarchical cluster analysis (CLA) and principal component analysis (PCA), were performed to identify protein molecular structural differences among the grains. The results revealed that CDC Seabiscuit contained greater (P < 0.05) protein structural Amide I and II than CDC Nasser and CDC Meredith, while the greater (P < 0.005) structural Amide I to II area and height ratios was detected in CDC Meredith. New oat grains had greater (P < 0.05) β-sheet height than barley grains, however, there was no difference in α-helix to β-sheet ratio values among the varieties. In conclusion, CDC Nasser and CDC Meredith had no difference in protein molecular structural features, while CDC Seabiscuit contains different protein structural characteristics as compared to CDC Meredith grain. The molecular structure features are highly associated with physiochemical and nutrient profiles in grains, which indicate that it also affect nutrient utilization and availability.     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

面向养殖水体分布差异的COD光谱法检测研究

面向养殖水体，传统光谱法对化学需氧量（Chemical Oxygen Demand，COD）检测模型构建的基础：源域（现有样本库）与目标域（检测地水体）间光谱数据独立同分布。但是当源域与目标域分布间存在差异时，由源域得到的低误差模型常在目标域上表现下滑。针对该问题，提出面向UV Vis光谱的域对抗训练网络（DAUVwpNet），将分布不同的源域和目标域数据映射至相同分布的特征空间中，使其在该空间的分布距离尽可能接近，从而在特征空间中对源域训练的目标函数也可以迁移至目标域上，以降低模型在目标域的误差。试验表明：面向同一批测试数据，DAUVwpNet的预测误差为0.78，要低于传统模型的预测误差（0.85）；DAUVwpNet预测值与实测值间相关系数为0.95，要高于传统模型的相关系数（0.89）。表明了该网络能够较好对齐两域特征空间数据分布，降低因分布差异带来的COD检测误差。     

配料混合试验的设计和分析

系统地介绍了配料混合试验的基本概念．指出了混合试验与常规因子试验的区别。提出配料混合试验的3种设计，即顶点和边界点设计、矩心设计和极顶设计。以2个实例演示了混合试验资料的统计方法，包括回归和方差分析．驻点分析和等值线分析。     

肾俞穴,后三里穴与子宫相关的神经基础

应用辣根过氧化物酶（ＨＲＰ）法和荧光素双标记法，研究了家兔子宫与肾俞穴、后三里穴的外周神经联系。结果显示，支配子宫与肾俞穴、后三里穴的初级传入神经元及交感节后神经元分布节段部分重叠，在腰１（Ｌ１）脊神经节有少量的荧光素双标记细胞，约占该节荧光素标记细胞总数的２．６％，表明Ｌ１脊神经节有一些初级传入神经元，其周围突分支分布于肾俞穴和子宫，提示子宫与肾俞穴除通过中枢途径联系外，还可通过外周突分支投射的感觉神经元直接相联系；在胸１２～荐３（Ｔ１２～Ｓ３）交感神经节出现大量分别支配子宫和后三里穴的标记细胞，这两种细胞紧密相邻，提示支配子宫和后三里穴的交感节后神经元之间，可能还存在直接或间接联系。上述结果为穴位－内脏相关机理提供了形态学证据。     

蓝舌病灭活疫苗免疫羊ELISA和琼扩监测比较试验

用19只绵羊（注苗前BTV琼扩和ELISA均为阴性）注射BTV灭活苗后7～139天的血清进行BTVELISA和琼扩平行试验，结果表明，注苗后7～30天11只试验丰ELISA阳性，注苗40天13只羊ELISA阳性，注苗90天以后全部试验羊只均为ELISA阳性，而琼扩试验则始终为阴性结果。     

单克隆抗体ELISA检测犊牛粪便中冠状病毒抗原

用两株单克隆抗体混合后包被ＥＬＬＩＳＡ微孔板，加牛冠状病毒抗原，加经白陶土处理过的兔抗牛冠状病毒免血清，再加上辣根氧化物酶标记的羊抗兔ＩｇＧ抗体，加底物质显色间接ＥＬＩＳＡ检测牛冠状病毒抗原获得成功。用此方法检测细胞培养的牛冠状病毒上清液，其敏感度高出血凝试验（ＨＡ）８倍，从武汉市附近３个奶牛场采集犊牛腹泻粪便１０５份，用单抗ＥＬＩＳＡ检测牛冠状病毒抗原，共检出阳性３６份，并与血凝及血凝抑制试验，