鲆鲽类主要病原菌抗血清的制备及应用

用甲醛灭活8种鲆鲽类主要病原菌,鳗弧菌(Vibrio anguillarum)、创伤弧菌(Vibrio vulnificus)、哈维弧菌(Vibrio harveyi)、溶藻弧菌(Vibrio alginolyticus)、嗜水气单胞菌(Aeromonas hydrophila)、豚鼠气单胞菌(Aeromonas caviae)、迟缓爱德华菌(Edwardsiella tarda)、鮰爱德华菌(Edwardsiella ictaluri),制备成灭活疫苗后肌肉注射大菱鲆获得各病原菌抗血清。结果显示,抗鳗弧菌血清、抗哈维弧菌血清、抗豚鼠气单胞菌血清、抗鮰爱德华菌效价为1:6 400,抗创伤弧菌血清、抗溶藻弧菌血清、抗嗜水气单胞菌血清效价为1:12 800,抗迟缓爱德华菌血清效价为1:102 400。应用ELISA分析各抗血清与8种病原菌及迟缓爱德华菌蛋白的免疫交叉反应,结果显示,各病原菌与其本身的抗血清反应最为强烈,而与其他抗血清间有程度不等的交叉反应;菌蛋白与其相应的菌抗血清反应最为强烈,与其他病原菌抗血清反应较弱。本研究所制备的病原菌抗血清效价较高,且具有特异性,可应用于鲆鲽类病原菌的检测。     

海洋有益菌的筛选与鉴定

从健康的斑节对虾（Penaeus monodon）幼体及其养殖环境中分离到213株海洋细菌,采用十字叉划线法和琼脂扩散法进行体外拮抗试验,从中筛选到5株对水产养殖动物病原哈维氏弧菌（Vibro harueyi）和鳗弧菌（Vibrio anguillarum）等菌株生长有抑制作用的海洋细菌。根据其形态特征以及生理生化反应特征,初步鉴定为交替单胞菌属（Alteromonas）,其中菌株A18鉴定为橙色交替单胞菌（Alteromonas aurantia）。     

食品中志贺氏菌快速检测免疫磁分离样品前处理技术研究

采用Protein A磁珠与志贺氏菌混合血清制备抗志贺氏菌多靶标免疫磁珠，建立了25 mL志贺氏菌免疫磁分离体系，并通过测定代表性食品(牛奶、水果、面包)中志贺氏菌生长曲线，设计食品样品前处理可行性技术方案，与环介导等温扩增(LAMP)检测技术整合进行了食品中痕量志贺氏菌快速检测验证。结果显示：多靶标免疫磁珠能同时捕获福氏、宋内氏、鲍氏、痢疾氏等4种志贺氏菌，捕获效率为35%～94%,非特异性小于1%(金黄色葡萄球菌10%除外),1～2×10¹ CFU志贺氏菌在25 mL纯牛奶、25 g梨/面包+25 mL志贺氏菌增菌肉汤基础中，通过6 h培养+免疫磁分离的策略可获得10² CFU/mL以上的志贺氏菌，将志贺氏菌检测样品前处理时间由48 h缩短至8 h内，并能够满足后续检测技术检出限的需求，实现了食品中志贺氏菌的快速检测。     

大菱鲆病原鳗弧菌生物学及分子特征研究

从山东沿海养殖发病大菱鲆分离了5株鳗弧菌。5株菌对大菱鲆有较强的致病性,LD50范围为2.32×101～4.03×102cfu/g鱼;5株菌分别属于O1、O2、O3血清型,菌株之间的16SrRNA基因序列相似性范围97.1%～99.9%;与国外报道的鳗弧菌的序列相似性范围为89.7%～99.9%。5株菌的生理生化特征与鳗弧菌标准菌株一致,能产生多种胞外酶和溶血素;都含有金属蛋白酶基因和溶血素基因,其中3株菌各含有一个67kb大质粒。病原菌对青霉素、苯唑青霉素、氨苄青霉素、先锋霉素Ⅳ、先锋霉素Ⅴ、先锋霉素Ⅵ、链霉素、强力霉素、洁霉素、万古霉素、灭滴灵等11种常用抗菌药物有抗性,只对新生霉素、呋喃妥因、利福平、新霉素等4种药物敏感。     

应用间接ELISA技术快速检测花鲈病原菌-鳗弧菌

以花鲈（Lateolabrax japonicus)弧菌病的病原菌－鳗弧菌（Vibrio auguillarum)W-1为抗原,制备兔抗血清;利用辣根过氧化酶标记的羊抗兔血清（HRP－IgG)为酶标二抗,建立了检测鳗弧菌的间接ELISA快速检测法。结果表明,应用间接ELISA技术检测鳗弧菌有较高的灵敏度,最低的检测量为10^5个／ml,即10^4个／孔。同时交叉反应表明其具有较高的特异性。对46份人工感染后的花鲈组织样品,包括肌肉、鳃、肠、肝、肾等组织的检测表明,阳性检测率为76．1％。     

鳗弧菌pEIB1质粒缺失株MVAV6201的筛选和三价铁源供给的优化

以鳗弧菌Vibrio anguiUarum MVM425(01型)为出发菌株,通过变温培养诱变筛选得到pEIB1质粒缺失株MVAV6201。通过与野生株MVM425的摇瓶培养对比发现,Fe^3 营养的摄取是质粒缺失株MVAV6201生长的瓶颈。MVAV6201对Fe^3 的吸收有赖于外界Fe^3 营养的供给形式。柠檬酸铁铵能有效增强MVAV6201对三价铁营养的吸收,促进菌体细胞的生长。柠檬酸铁铵饱和浓度为0．4mmol／L。     

鳗弧菌金属蛋白酶单克隆抗体的制备及鉴定

鳗弧菌M3胞外产物经纯化得到分子量为36.5kDa的金属蛋白酶，浓缩后作为抗原免疫BALB/C小鼠，经细胞融合及筛选。得到三株稳定分泌抗金属蛋白酶的杂交瘤细胞株。分别命名为P2B1，P4A3和P4B1。亚型鉴定结果表明，三株细胞株产生的单抗均为IgG3型。蛋白质印迹（Wes tern Blot)分析显示，所得到的单抗只与分子量为36.5kDa的金属蛋白酶反应，具有较高特异性。三株单抗均能有效抑制蛋白酶活性。     

大菱鲆免疫球蛋白M(IgM)单克隆抗体的制备与特性鉴定

应用细胞工程技术研制大菱鲆免疫球蛋白M(immunoglobulin M,IgM)的单克隆抗体并分析其免疫学特性。小鼠骨髓瘤细胞NS0与经IgM免疫的BALB/C小鼠脾细胞融合,经过反复有限稀释法克隆,筛选获得4株抗大菱鲆IgM的单克隆抗体杂交瘤细胞株,分别为B1D1、D5C2、E1B2和F4A1。经小鼠腹水扩大生产后,单抗效价为1∶1.024×106检测灵敏度为32 ng/mL。Western blot分析表明,获得的单抗与IgM重链区特异性结合。交叉结果显示,单抗与大菱鲆血清呈强阳性反应,与褐牙鲆、红鳍东方鲀、许氏平鲉、六线鱼、鲈鱼均呈微弱阳性反应,而与半滑舌鳎、鲤鱼、鲫鱼、草鱼、鳙鱼的血清无交叉反应。本研究制备的大菱鲆IgM单克隆抗体效价高、灵敏度高、特异性强,适合用于大菱鲆免疫学相关研究和生产实际应用。     

对抗氯霉素单克隆抗体4D10特性鉴定研究

目的对抗氯霉素单克隆抗体4D10进行纯化与特性鉴定,以获得可用于检测氯霉紊残留试剂盒的单克隆抗体。方法用两步硫酸铵法和G蛋白亲和层析法纯化单克隆抗体,用间接酶联免疫吸附试验、竞争ELISA等方法测定抗体效价、特异性、敏感性与亲和性以及抗体与其它氯霉素结构类似物的交叉反应率。结果纯化后抗体的纯度高达95％,对抗体进行相关性质的鉴定得出：4D10效价为1：2．56×105,其亲和力常数可达2×108M-1,该抗体与CAPBSA,CAPOVA有反应,与BSA、OVA无反应。与氯霉素琥珀酸钠、氯霉素反应,而与其它氯霉索结构类似物无交叉反应。结论抗氯霉索单克隆抗体4D10效价高、特异性强、亲和力高,可利用该抗体制备检测氯霉索残留的ELISA试剂盒。     

有益细菌A18在海湾扇贝(Argopecten irradians)育苗中的应用

在海湾扇贝D形幼虫期施用10^3,10^4,10^5CFU／ml三个浓度的有益细菌A18。经该菌处理后，扇贝幼苗的变态率与各个发育期的成活率均比加氯霉素组和未加任何处理的对照组高。其中以10^4CFU/ml处理的扇贝幼苗的变态率和成活率最高。对实验扇贝幼菌水体细菌总数和弧菌数分析发现，经10^5,10^4CFU/ml A18菌处理水体的细菌总数和弧菌数均比相应的对照组低。在扇贝育苗水体加入10^6,10^7,10^8CFU／ml的有益细菌A18，检测高浓度A18对扇贝幼苗的毒害作用，当该菌浓度在10^6CFU/ml时，经48小时扇贝幼苗成活率79％，高于对照组的68.5%；当浓度达10^7,10^8CFU/ml时，处理48小时后的扇贝幼苗死亡率分别为95.6%和100％。     

The Research of Anti-Wear Properties of Organomolybdenum Compounds in the Engine Oils

A four-ball tester was used to evaluate the anti-wear performance of three kinds of organomolybdemun compounds in the engine oils, i. e., molybdenum dialkyldithiophosphate (MoDDP), molybdenum dialkyldithiocarbamate ( MoDTC), and sulphur and phosphorus freeorganomolybdeum (Molybdate). The results indicate that a low concentration of MoDDP doesn' t improve the anti-wear properties of the commercial engine oils, but a high concentration of MoDDP can obviously improve the anti-wear properties and the load-carrying capacity of the engine oils. MoDTC doesn' t improve the antiwear properties of the engine oils, but worsens the anti-wear properties of the oils. Signifi can timprove ment of frictional and wear characteristics is obtained with Molybdate added in the commercial engine oils and the formulated oils.     

Certification Becomes Important for the I Quality; and Safety Supervision of the Chinese Liquor Sector

The end of 2007, over 200 unit products of more than 80 Chinese firms have passed the quality grade certification for liquor products. These products involve distilled spirits, beer, wine, yellow wine, fruit syrup wine and others, and cover over 80% of the national top-branded liquor products.……     

Deposition and Coating Properties on CVD Tungsten

Surface characterization and microstructure studies are performed on chemical vapor deposited (CVD) tungsten coating. There is about 2 μm thickness diffusion layer of tungsten in the molybdenum substrate. The thermal shock test shows tungsten coating has good adhesion with molybdenum substrate, but the elements of oxygen and carbon in the tungsten coating have the bad affection to the adhesion. The result of high-temperature diffusion experiment is the diffusion rate from molybdenum substrate to tungsten coating is faster.     

Analysis of marine hydrobiont lipid extracts as possible cryoprotective agents

We studied a combined effect of the cryoprotectants both of lipid and carbohydrate origin at the presence of DMSO or without it on cell viability and the RNA synthesis in the embryonic mollusc and echinoderm cell cultures obtained at different developmental stages. Cryoprotective properties of exogenous lipids correlated with their thermotropic behavior. Lipid extracts from marine hydrobiontes, which thermal transitions are characterized by relatively small part of calorimetric thermogram area lying at temperatures above 0 °C (6–18%), were found to possess a considerable cryoprotective activity. The results are discussed in relation to the prediction of the cryoprotective activity of lipid extracts for marine animal cells. In addition, it was found that marine invertebrate cells can be cryopreserved without DMSO at the presence of trehalose and lipid extracts of some marine hydrobiontes.     

Low-Temperature Properties of Silver

Pure silver is used extensively in the preparation of high-temperature superconductor wires, tapes, films, and other configurations in which the silver not only shields the superconducting material from the surrounding materials, but also provides a degree of flexibility and strain relief, as well as stabilization and low-resistance electrical contact. Silver is relatively expensive, but at this stage of superconductor development, its unique combination of properties seems to offer the only reasonable means of achieving usable lengths of conductor. In this role, the low-temperature physical (electrical, thermal, magnetic, optical) and mechanical properties of the silver all become important. Here we present a collection of properties data extracted from the cryogenic literature and, to the extent possible, selected for reliability.     

Early ironwork in Biscay: Survey,excavation, experimentation and materials characterization. An integral study of the mountainside ironworks (ferrerías de monte or “haizeolak”)

ABSTRACT

The production of iron in Biscay (Basque Country, Spain) has been a hallmark of this territory of the North of the Iberian Peninsula throughout its history. The Archaeology team of the Basque Country Mining Museum has spent 13 years, so far, studying the origins of this activity through the evidences left at the old pre-hydraulic iron production workshops known as mountainside ironworks (ferrerías de monte) or haizeolak. It has catalogued 170 archaeometallurgical sites of this type in Biscay, and the archeological excavation of some of them has led to the knowledge of the different stages of the production process that took place in these workshops. The team has also experimentally reproduced the work carried out in a mountainside ironworks from the High Middle Ages, and it has currently started the characterization process of this type of activity through the study of the collected materials.     

Mechanism Study on the Wear of CrAlTiN Coated High-Speed Steel Twist Drills Under Dry Cutting Conditions

As demands about environment protection are growing up, dry cutting technology is getting more and more concerns from all over the world. Main works performed here are study on dry cutting performances and wear mechanisms of M2high-speed steel (HSS) twist drills with CrAlTiN multicomponent coatings, which was deposited using magnetron sputter ion plating system, in drilling 45 and 30CrMnSiA steel, and their comparisons to those in drilling the same steel but using monolayer CrN and commercial TiN coated drills. Drilling performances of drills are evaluated mainly through the measurements of width on outer corner flank land and the cutting forces. Results show that performances of CrAlTiN coated drills are better than those of monolayer CrN and TiN coated drills. In drilling 45 and 30CrMnSiA steel, the average tool life of the CrAlTiN coated drills are 17.2 and 11.8 times higher than those of the uncoated drills. Observing wear of the drill with scanning electronic microscope, results show that there is no crack or spallation of the CrAlTiN coatings in wear zones. Main mechanism of the wear here is adhesion.     

Study on Bond Ability of Arc-Spraying Coatings with Different Surface Pretreatment on Cast-Iron

Arc spraying coatings are widely used in various applications, but uncommon in cast iron substrate. Different surface pretreatment technology is tested on substrates of gray cast iron. Surface roughness and residual stress were measured by TR200 and X-ray diffraction analyzer. Influence of different surface pretreatment methods ( dry blasting and fusebond) on roughness and residual stress was analyzed. The arc-sprayed coatings of wire 3Cr13 (φ2mm) on gray cast iron substrate is studied. The microstructure and interface of bonding layer were observed by SEM. The bond strength was taken by tensile test. Results show that bond strength with grit blasting is higher than fuse-bond; it is feasible to make wire 3Cr13 coating with arc spraying on cast iron substrate roughened by grit blasting.     

Structural Characteristics of TiO2 Ceramic Coating by Micro-Plasma Oxidation

TiO2 ceramic coatings with thickness of 20 μm were formed on the surface of pure titanium by micro-plasma oxidation. Their micro-structures were investigated by by using X-ray diffraction and their surface images were detected by using scan electronic microscope. There were three kinds of TiO2 coatings, pure anatase type TiO2 phase, mixed phases consisted of rutile type TiO2 phase and anatase type TiO2 phase, pure rutile type TiO2 phase. The coating surface with the pure anatase type TiO2 phase is rough, while the coating surface with the pure rutile type TiO2phase is smooth. The upper coating surface with the mixed type TiO2 phases is anatase type TiO2 structure and the subsurface of the TiO2 coating is rutile type TiO2structure.     

Evaluation of fatigue life of steel using steel grain size

The insightful discussion of the relationship between the construction of metallographic and fatigue life and proposition of mathematical relationships describing this relationship was conducted in the paper. This paper presents a method for estimating the fatigue life, based on the construction of the microstructural material taking into account the grain size and the participation of ferrite and pearlite phase.