液体农药制剂中烷基酚及其聚氧乙烯醚类助剂的高效液相色谱分析

建立了一种测定6类液体农药制剂中烷基酚（alkylphenols,APs）及其聚氧乙烯醚类（alkylphenol ethoxylates,APEOs）助剂的高效液相色谱-荧光分析方法。乳油、水剂、水乳剂、悬浮剂、可溶液剂和微乳剂样品均经甲醇超声辅助提取,稀释后过聚丙烯膜,采用Poroshell 120 EC-C18（4.6 mm×100 mm,2.7 μm）分离,V（甲醇）:V（乙腈）:V（水）=75.5:6.5:18.0三元流动相等度洗脱,荧光检测器检测,外标法定量。结果表明:在0.05~10或0.1~20 mg/L范围内,APs和APEOs的峰面积与其质量浓度间呈良好线性关系（R2均大于0.999 8）;在添加水平范围内（APs:0.5、2、5 mg/g;APEOs:1、5、25、50 mg/g）,6类液体农药剂型中APs和APEOs的平均回收率为73%~110%,相对标准偏差为0.6%~14.9%,方法的定量限在0.5~1 mg/g。该方法简单、快速、准确,适用于不同液体农药制剂中烷基酚及其聚氧乙烯醚类助剂的测定。     

茭白和甜玉米对甘蔗二点螟生长发育的影响

[目的]明确茭白和甜玉米对甘蔗二点螟生长发育的影响,为开展甘蔗二点螟人工大量饲养及防控技术研究提供科学依据.[方法]在温度(26±1)℃的空调房中,挑取初孵二点螟幼虫单头接入培养皿内,分别饲以茭白茎段和甜玉米粒块,每3d更换1次食料,每天定时观察记录幼虫的发育情况和存活数量,幼虫化蛹后第2d观察记录其性别及称量体重,记录幼虫化蛹、蛹羽化、成虫死亡日期,计算各虫态的发育历期、存活率及雌、雄性比.每处理重复50皿,试验共培养3批次.[结果]取食茭白和甜玉米的二点螟幼虫平均历期雄性分别为22.54和26.29 d,雌性分别为24.69和29.77 d,两者均达极显著差异水平(P<0.01);取食茭白和甜玉米的二点螟蛹平均历期分别为8.68和8.94 d,成虫平均寿命分别为4.12和3.90 d,两者均差异不显著(P>0.05);蛹平均体重分别为0.0736和0.0651 g,差异达显著水平(P<0.05);初孵幼虫饲养至蛹时的平均存活率分别为49.30%和38.00%,饲养至成虫羽化时的平均存活率分别为46.00%和36.00%;雌雄性比分别为0.70和0.55.[结论]甘蔗二点螟室内取食茭白较甜玉米更适合其个体的生长发育.     

Mechanical properties of flax fibres treated by alkaline,enzyme and steam-heat

In recent years there has been interest in using flax fibres to produce composites because of a number of attributes, including low density, biodegradability and high mechanical properties. It was found that treatment of flax fibres may be required to improve the bond quality with a resin. These treatments also have an impact on the properties of the fibres themselves. The objective of this project was to evaluate the impact of three treatment methods on the mechanical properties of flax fibres. The three treatment methods were alkaline, enzyme and steam-heat. After treatment, flax fibres were tested in tension using a universal test machine. Results showed that tensile strength and Young's modulus of flax fibre can be enhanced significantly by the three treatment methods, compared with untreated flax fibres. Enzyme treatment was shown to be the best approach to improve mechanical properties of flax fibre than alkaline and steam-heat treatment.     

高置式大容量毒饵站对东北农田害鼠的防治效果初探

毒饵站是化学防鼠常用设施,可以减少非靶标动物的误食风险。目前常用毒饵站容量小、毒饵易浪费、添加毒饵不便的缺点限制了其在东北地区的大规模应用。本研究针对东北农业环境特点,设计了一种新型高置式大容量毒饵站,在黑龙江了进行为期1年的防效试验,并测试其作用范围。结果表明,这种毒饵站每年毒饵用量2.8~3.8kg,鼠密度控制率可长期保持在75%以下;其有效覆盖面积为1.1hm2,最佳防治效果的覆盖面积为0.5hm2。这些结果说明这种大容量高置式毒饵站具有毒饵添加便捷、无浪费的优点,在东北地区防治经济效益远远大于其他传统小容量毒饵站,值得在当地应用和推广。     

磁性分子印迹聚合物的制备及对食品中氯霉素残留的检测

将磁性分离技术和表面分子印迹技术相结合,首先通过化学改性的方式在Fe₃O₄磁性纳米粒子表面接枝双键,以氯霉素(chloramphenicol,CAP)为模板分子、甲基丙烯酸(methacrylic acid,MAA)为功能单体、乙二醇二甲基丙烯酸酯(ethylene glycol dimethacrylate,EDGMA)为交联剂、偶氮二异丁腈(azobisisobutyronitrile,AIBN)为引发剂,采用悬浮聚合法合成CAP的磁性分子印迹聚合物微球(magnetic molecularly imprinted polymer,MMIP),以及对应的非印迹微球(magnetic molecularly non-imprinted polymer,MNIP)。在表征试验中,用傅里叶变换红外光谱仪对每一步合成的产物进行红外光谱检测,用扫描电子显微镜对MMIP和MNIP的微观形态进行观察,用振动磁强计测定磁性纳米粒子和MMIP的饱和磁强度。对MMIP和MNIP的吸附性能进行研究,将MMIP作为固相萃取剂应用于实际样品检测,进行方法学考查。试验结果表明,合成的磁性分子印迹聚合物微球直径400~700 nm,分散性较好,在溶剂中可在外加磁场作用下快速分离。MMIP的最大表观吸附容量可达29.18 mg·g^-1,具有良好的选择识别性能。MMIP作为固相萃取剂在对实际样品进行检测时回收率(86.30%~94.21%)、精密度(RSD≤1.53%)、稳定性(RSD≤1.87%)均良好,且具有较低的检测限(3.0 μg·kg^-1)。将MMIP作为固相萃取剂用于食品中残留氯霉素的分离检测具有良好的效果。     

15N-labelled fertiliser recovery by maize (Zea mays L.) and leaching of nutrients as influenced by oil palm empty fruit bunch biochar in a mini-lysimeter under controlled tropical environment

Converting oil palm empty fruit bunches into biochar is an alternative waste management method and has strong potential to improve N fertiliser use efficiency in agriculture. The aim of this study was to determine the effectiveness of oil palm empty fruit bunch biochar (EFBB) in improving recovery of ¹⁵N-labelled nitrogen fertiliser by maize (Zea mays L.) and leaching of mineral N and K. An experiment was conducted in a mini-lysimeter system with randomised complete block design layout and six replications under controlled environment in a rain shelter. Each mini-lysimeter was filled with 20 kg of sandy loam soil before adding EFBB (0, 5, 10 and 20 Mg ha^?1). The N source used was (¹⁵NH₄)₂SO₄ at 80 kg N ha^?1 (2 at% ¹⁵N excess). Maize was irrigated to induce leaching every 4 days. Maize plant and soils were sampled 58 days after sowing (tasselling stage). Application of EFBB significantly reduced cumulative leachate volume and mineral N leaching. Soils applied with EFBB significantly improved ¹⁵N fertiliser recovery in maize and dry matter weight. This study shows that EFBB has the potential to be applied on highly weathered acidic soil as an amendment to improve fertiliser efficiency and crop growth.     

新城疫病毒LaSotaC5株感染性克隆的构建及病毒拯救

将本实验室保存的一株LaSota病毒用有限稀释法接种鸡胚进行纯化,连续传代5次后筛选到1株高血凝效价的纯培养克隆株,命名为LaSotaC5,并对其进行全基因组测序,克隆株与亲本株在生物学特性与基因序列上都存在一定的差异。参照LaSotaC5株全基因序列单酶切位点,用RT-PCR的方法将基因组分8段扩增,按照病毒基因组的结构顺序,将克隆片段定向插入到TVT转录载体中,成功构建含有病毒全长eDNA的转录载体TVT．LaSotaC5。将TVT-LaSotaC5与辅助质粒pCI-NP、pCI-P和pCI—L共转染BSR—T7／5细胞,成功拯救出了具有感染性的LaSota株新城疫病毒。病毒的成功拯救为后续基因功能和疫苗载体开发等方面的研究提供了平台。     

常见耐药基因多重PCR方法的建立及用于禽致病性大肠杆菌耐药性监测

本研究旨在建立β-内酰胺类、四环素类、氨基苷类、酰胺醇类、磺胺类抗菌药物耐药基因的多重PCR检测方法,用于耐药基因的快速检测。根据GenBank公布的上述5类抗菌药物的耐药基因序列,设计17对特异性引物。通过优化PCR体系和反应程序,建立4组耐药基因（cat+floR+tetB+tetC;dfrA12+sul2+sul1+bla_CTX-M+bal_TEM-1;aac3+aph3+aadA1+strB;tetA+cmlA+strA+sul3）的多重PCR反应体系。然后,检测多重PCR方法的特异性和敏感性。利用建立的多重PCR方法检测42株禽致病性大肠杆菌的耐药基因,同时,检测其耐药性,比较分析耐药基因和耐药表型之间的相关性。结果显示,建立的4组多重PCR体系可有效扩增出17个耐药基因片段,测序结果表明特异性较好。敏感性结果表明,4组多重PCR体系的菌液敏感性分别为10³、10⁴、10⁴和10⁵CFU。禽致病性大肠杆菌分离株的耐药基因多重PCR和单重PCR检测结果一致,其携带的耐药基因和耐药表型的符合率为92.86%。本研究建立的耐药基因多重PCR方法能简便、快速地检测常见的耐药基因,可用于耐药基因的传播、流行调查。     

ZB transposon and chicken vasa homologue (Cvh) promoter interact to increase transfection efficiency of primordial germ cells in vivo

ABSTRACT

1. In order to increase the efficiency of generating transgenic chicken, this trial focused on two points: primordial germ cells (PGCs）transfection in vivo and a germline-specific promoter.

2. In order to transfect PGCs in vivo, two plasmids (pZB-CAG-GFP, pCMV-ZB）were co-injected into chicken embryos via the subgerminal cavity at Hamburger and Hamilton (HH) stage 2–3 or via blood vessel at HH stage 13–14. Results showed that the percentage of GFP+ embryos, viability and hatching rate of embryos injected at HH stage 13–14 were significantly higher than that at HH stage 2–3.

3. Two plasmid transposon systems were used for chicken embryo micro-injections. The donor plasmid, with a green fluorescent protein (GFP) reporter gene, was mediated by the ZB transposon. The helper plasmid was a transposase expression vector driven by the promoter of the chicken vasa homologue (Cvh) gene or Human cytomegalovirus (CMV) promoter. Results showed that 60.98% of gonads in Cvh group expressed GFP, which was 52.50% higher than seen in the CMV group. Only gonad tissue from the Cvh group showed any GFP signal, whereas both gonads and other tissues in the CMV group showed green fluorescence.

4. The data suggested that ZB transposon-mediated gene transfer was efficient for transfecting PGCs in vivo; the Cvh promoter drove the transposase gene specifically in the germline and increased the efficiency of germline transmission. Blood vessels injection at HH stage 13–14 may be a more efficient route for PGCs transfection in vivo.