Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

白斑综合征病毒环介导等温扩增快速检测方法的建立

根据对虾白斑综合征病毒(WSSV)囊膜蛋白VP28基因保守序列,利用Primer Explorerv 4.0软件设计了4条引物,建立了白斑综合征病毒环介导等温扩增快速检测方法,对反应温度和反应时间等参数进行了优化,同时将建立的LAMP检测方法与巢式PCR进行了比较分析。结果表明,LAMP最适反应在64℃恒温条件60min内完成,凝胶电泳呈现梯型条带;反应体系中添加SYBR Green I荧光染料后,绿色的阳性结果明显区别于橙色阴性结果。LAMP方法的最低检出限为100拷贝/μL,灵敏度较巢式PCR高100倍,而且LAMP方法在1h内即可完成检测,操作简单,无需复杂仪器,肉眼可直接观察检测结果。用建立的LAMP方法对临床发病南美白对虾样品进行了检测,结果表明,LAMP方法适合对虾白斑综合征病毒的现场快速检测。     

我国食品中农药残留研究进展

随着人们安全健康意识的提高,食品中农药残留问题更加受到重视。为此,对样品预处理研究进展及农药残留快速检测技术研究进展进行综述,并简单介绍各种方法的优缺点。     

基于改进Hough变换的农田作物行快速检测算法

选取苗期农田作为研究对象,采集了包含行栽作物和土壤背景的农田图像,针对现有作物行定位方法易受外界干扰和处理速度较慢的不足,提出将投影法和直接Hough变换法相结合检测作物行的算法.采用2G-R-B 法和OTSU法将图像二值化,通过快速中值滤波算法去除噪声,再利用垂直直方图投影将图像进行水平条划分获取作物垄平均定位点,最后通过Hough变换检测垄定位点,得到作物行中心线.试验结果表明:基于垂直直方图投影的Hough变换检测作物行中心线的算法在保证高定位精度的同时,算法处理速度比直接Hough变换检测法提高了3倍,得到的定位基准线能代表作物行走向.     

采用施奈德重合器实现馈线自动化的实践研究

针对基于传统重合器的馈线自动化原理及存在问题,提出基于施奈德重合器的馈线自动化方案。以大连庄河炮台变电所石山线-龙王庙变电所水泥线为例,分析施奈德重合器的馈线自动化工作原理,指出该馈线自动化系统在实践中的优点和不足。     

基于阵列芯片现场高灵敏同步快速检测粮油中氨基甲酸酯类农药残留混合污染

The quality and safety of agricultural products is the technique focus of the strategy of rural vitaliza⁃tion. Carbamate pesticide residues are seriously threatening the safety of agricultural product and human life and health. Nowadays, the common detection methods are instrument-based for carbamate pesticide residue, which dis⁃ satisfy the request of an on-site rapid, sensitive and simultaneous detection for multiple carbamate pesticide resi⁃ dues. Based on the home-made monoclonal antibodies against carbofuran and carbaryl respectivity, a microarray for rapid and simultaneous detection of multiple carbamate pesticide residues was developed within 30 min. Under the optimized conditions, the limits of detection（LODs）were found to be 0.11 and 0.09 ng/g, while the linear ranges were 0.33-3000 and 0.18-1500 ng/g for carbofuran and carbaryl respectively. By using spiked peanut samples, it recorded recoveries of 88.4%-98.3%, 90.1%-118.8% for carbofuran and carbaryl respectively. The RSD of intraarray were found at 9.0%, 3.5%, while the RSD of inter-array were recorded at 12.3%, 16.3% respectively. The de⁃ tection results of the microarray were highly consistent with that of HPLC-MS/MS. This proposed microarray provid⁃ ed an on-site high-sensitivity, simultaneous and rapid detection of carbofuran and carbaryl, and could be widely used in the fields of agricultural product quality safety, food safety and environmental monitoring.     

槟榔黄化病病原及检测方法研究进展

黄化病是一种严重威胁槟榔生产种植的毁灭性传染性病害,目前主要在印度和我国槟榔主产区发生。本文从非生物与生物因素两方面对槟榔黄化病病原认识过程及检测方法进行详细的综述。     

基于蚁群优化算法的图像边缘检测

图像边缘携带了图像的大部分主要信息。通过对图像进行边缘检测不仅能有效地提取图像信息降低计算的复杂度而且是图像测量、图像分割、图像压缩、模式识别等图像处理的基础。本文尝试将蚁群优化算法（Ant Colony Optimization, ACO）用于图像边缘检测,通过选取经典house图像和SAR机场图像设置阈值进行自适应边缘提取,实现了边缘的精确检测。实验结果显示,该算法能够有效地提取图像目标的轮廓信息,很好保持图像纹理,具有理想的抗干扰性能,保证了检测结果的准确性。     

基于多结构鲁棒估计的虹膜外边缘定位方法

在Adaboost检测人脸区域的基础上,提出了基于多结构鲁棒估计的虹膜外边缘定位方法．通过对200幅单人脸的图像数据集的仿真实验,结果表明,本方法相比于传统的RANSAC模型生成算法以及Hough变换等方法,能得到更精确的定位结果,而且能加快有效的模型生成,虹膜外边缘定位精度达到94％．