文蛤副溶血弧菌间接 ELISA检测技术的研究

从患病文蛤组织内分离出一株细菌,回归感染试验证明是文蛤的致病菌,经鉴定为副溶血弧菌,灭活后(56 ℃,1 h)制成菌苗,用该菌苗制备兔抗血清,以辣根过氧化酶标记的羊抗兔血清(HRP-IgG)为酶标二抗,对副溶血弧菌进行间接ELISA快速检测.结果表明:应用间接 ELISA 技术检测副溶血弧菌具有较高的灵敏度,其最低检测极限为 1 × 105 cfu / mL.患病文蛤内脏团中副溶血弧菌的检出率为 80 %,无病症带菌文蛤中的检出率为 15 %,海水中副溶血弧菌的浓度低于最低检测极限.表明间接 ELISA检测法不仅可以用于发病文蛤的快速检测,而且能够检测出无病症的带菌的文蛤,为有效预防副溶血弧菌病的暴发提供了检测手段.     

大黄鱼病原菌——溶藻弧菌的EuSA快速检测研究

以灭活溶藻弧菌(0.05%甲醛,30℃,8h)制成菌苗,注射免疫实验兔获得抗血清.该抗血清经吸附后与副溶血弧菌、河流弧菌等10株对照菌株无交叉反应.用该抗血清进行溶藻弧菌ELISA检测,其最低检测极限为97000cfu/ml.将该方法应用于养殖现场,检测结果为海水中溶藻弧菌浓度均低于最低检测限、外观健康大黄鱼肝脏中溶藻弧菌的阳性检出率为20%、患病大黄鱼的阳性检出率为80%,表明ELISA法不仅可以用于诊断大黄鱼的溶藻弧菌病,也可以检测无病症带菌大黄鱼.     

大黄鱼病原菌──溶藻弧菌的ELISA快速检测研究

以灭活溶藻弧菌（0.05%甲醛，30℃,8h）制成菌苗，注射免疫实验兔获得抗血清。该抗血清经吸附后与副溶血弧菌、河流弧菌等10株对照菌株无交叉反应，用该抗血清进行溶藻水中溶藻弧菌浓度均低于最低检测限、外观健康大黄鱼肝脏中溶藻弧菌的阳性检出率为20%、患病大黄鱼的阳性检出率为80%，表明ELISA法不仅可以用于诊断大黄鱼的溶藻弧菌病，也可能检测无病症带菌大黄鱼。     

大黄鱼病原副溶血弧菌单克隆抗体制备及其应用

用甲醛灭活副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(V.alginolyticus)制成免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,获得1株特异的针对副溶血弧菌的单克隆抗体,命名为D6F3H5。腹水及培养上清液抗体的ELISA效价分别为:1∶5 120和1∶1 280,该单克隆抗体与其它细菌没有明显的交叉反应。利用该单克隆抗体和兔抗副溶血弧菌多克隆抗体,建立了检测副溶血弧菌的双抗体夹心ELISA方法。该方法对副溶血弧菌的最小检出量为5×104个/mL。用双抗体夹心ELISA检测大黄鱼(Pseudosciaena crocea)样品,14尾患病大黄鱼中有11尾检出副溶血弧菌,而10尾健康大黄鱼都没有检出副溶血弧菌。由此可见,本试验制备的高特异性的抗副溶血弧菌单克隆抗体,可以用于患病大黄鱼副溶血弧菌的快速诊断。     

副溶血弧菌胶体金快速检测试纸的研制及应用

副溶血弧菌(Vibrio parahaemolyticus)是海水养殖动物弧菌病的主要病原菌之一。为实现对该菌的现场快速检测,本文以制备的兔抗副溶血弧菌多克隆抗体作为金标抗体,提取副溶血弧菌的外膜蛋白、鞭毛蛋白、胞外产物和全菌蛋白,以不同浓度的4种抗原蛋白作为4条检测线(T1~T4),特别在T4线设置全菌蛋白,基于竞争免疫层析原理研制出副溶血弧菌胶体金快速检测试纸。使用该试纸检测了副溶血弧菌(V.parahaemolyticus)、鳗弧菌(V.anguillarum)、溶藻弧菌(V.alginolyticus)、哈维氏弧菌(V.harveyi)和鱼肠道弧菌(V.ichthyoenteri),检测结果表明其能够准确鉴别副溶血弧菌感染,排除其他菌的交叉反应干扰。副溶血弧菌快速检测试纸的最低检测限为5×105 cfu·mL^-1,感染副溶血弧菌的牙鲆组织的检测结果与ELISA结果一致。本文研制的试纸具有较高的特异性与准确性且操作简单,为现场快速检测副溶血弧菌提供了有力工具。     

大黄鱼溶藻弧菌LPS的间接ELISA检测

用酚 水法从20dm3溶藻弧菌培养液中分离得到脂多糖(LPS)97.6mg,以0.5mg cm3的LPS溶液通过耳缘静脉注射免疫实验兔,经过3次加强注射后从颈动脉放血制备抗血清.该抗血清的效价为1∶2560,对副溶血弧菌等10株细菌的交叉反应效价都较低.用免疫吸附过滤法去除血清中的非特异性成分.用吸附后的抗血清进行溶藻弧菌间接ELISA检测,其检测限为97×104个 cm3.以ELISA进行大黄鱼体内溶藻弧菌检测.结果表明:用LPS抗血清可以较为灵敏地检测大黄鱼体内的溶藻弧菌,该方法不仅可以用于诊断大黄鱼的溶藻弧菌病,也可以检测无病症带菌大黄鱼.     

溶藻弧菌的间接荧光抗体快速检测

建立了大黄鱼病原溶藻弧菌的间接荧光抗体检测方法，溶藻弧菌的特异性抗血清由新西兰兔制备，试管凝集法测定抗血清及交叉反应效价，吸附离心法去除交叉反应，荧光抗体为异硫氰酸荧光素标记的羊抗兔免疫球蛋白，现场检测对具典型弧菌病表症大黄鱼病原检出率为100％，表观健康大黄鱼病原检出率为30％，养殖海水检测结果均为阴性，结果表明，间接荧光抗体检测法不仅可以用于已感染发病大黄鱼的快速检测，而且还能用于已感染未发病大黄鱼的检测。     

检测海洋弧菌的酶联免疫吸附试验研究

研究建立中国对虾病原菌┐副溶血弧菌（Ｖｉｂｒｉｏｐａｒａｈａｅｍｏｌｙｔｉｃｕｓ）的间接酶联免疫吸附试验（ＥＬＩＳＡ）快速检测技术。间接ＥＬＩＳＡ技术中的副溶血弧菌的特异抗血清由家兔制备，羊抗兔ＩｇＧ用碱性磷酸酶标记，酶的底物为对磷酸基硝基苯。将该技术标准化后，测定了抗血清与１３株其它副溶血弧菌菌株及２９株其它弧菌标准菌株的交叉反应，并进行了苗期对虾样品中副溶血弧菌的检测     

间接ELISA技术在病原性鳗弧菌SMP1快速检测中的应用

以大菱鲆(Scophthalmus maximus)出血症的病原菌——鳗弧菌(Vibrio anguillarum)SMP1为抗原,制备兔抗血清,测定了抗血清的工作浓度和对抗原的特异性,建立了在大菱鲆肝肾组织中检测SMP1的间接ELISA方法。SMP1人工感染大菱鲆,用建立的间接ELISA对大菱鲆肾脏和肝脏进行检测,结果肾脏的阳性检出率为87.7%,肝脏的阳性检出率为90%。该方法的建立有助于快速准确地鉴定由SMP1引起的大菱鲆疾病。     

中国对虾弧菌病的间接荧光抗体诊断技术研究

于１９９４年８月－１９９５年５建立中国对虾病原菌－副溶血弧菌的间接荧光抗体检测技术，其中副溶血弧菌的特异抗血清由家兔制备，羊抗兔免疫球蛋白用异硫氰酸荧光素标记，并以罗丹明标占血清白蛋白为背影染色。     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.     

Estimation of Sediment Properties during Benthic Impact Experiments

Sediment properties, such as water content and density, have been used to estimate the dry and wet weights, as well as the volume of sediment recovered and discharged, during benthic impact experiments conducted in the Pacific and Indian Oceans. The estimates show that the weights of dry (355-1332 t) and wet sediment (1651-4888 t) recovered during the experiments, are not only a function of the total duration and distance covered during the experiment, but also depend on water content and density of the sediment. Estimation of dry sediment and its volumetric ratio in the discharge, are key parameters for calculating the discharged volume. The estimated volume of wet sediment recovered (1427-4049 m 3 ) as well as discharged (2693-6951 m 3 ) during the experiments, provide important inputs to evaluate the sediment resettlement and migration. Using these estimates, the average depth of excavation on the seafloor, can also been calculated. However, the stages of sediment recovery and discharge are expressed differently in some of the experiments. In order to standardize these, different stages have been identified, and definitions of certain terms have been suggested, for use in the future. The methods of calculating different properties, as well as weight and volume of discharged sediment are described in the paper for use in other applications concerning deep-sea discharges.     

A Tidal Flow Model for the Gulf of Kachchh, India

A vertically integrated model has been used to study the tidal circulation and currents in the Gulf of Kachchh along the west coast of India. The model is fully nonlinear and uses a semiexplicit finite difference scheme to solve the basic hydrodynamic equations on a staggered grid. The model is forced by prescribing the tides along the open boundary of the model domain. The flow is simulated both with and without the presence of the proposed tidal barrage across the Hansthal Creek in the Gulf of Kachchh. The results show a considerable change in the behavior of the tidal flow in the presence of the barrage.     

利用中国GF-3卫星SAR和HY-2A卫星微波散射计的首次台风联合定量观测

<正>The typhoon,as a mature tropical cyclone that develops in the western part of the North Pacific Ocean with high wind speed and heavy rainfall,is one of the most lethal and costly of natural disasters for the densely populated countries of East Asia.It can be easily detected by space-borne sensors operated at microwave,visible or infrared bands(Liu et al.,2014).Synthetic Aperture Radar(SAR)is     

离子交换—原子吸收光谱法测定海水中痕量铜、铅、锌、镉、铁、锰

本文介绍了用国产D401型螯合树脂分离富集海水中铜、铅、锌、镉、铁、锰等痕量元素,并用原子吸收光谱仪测定其含量的方法。讨论了各元素的分离条件选择及干扰元素的影响,并与溶剂萃取法的结果作了比较。各元素检测的定量下限为:铜0.5μg/L、铅0.1μg/L、锌1.0μg/L、镉0.01μg/L、铁2.0μg/L、锰2.0μg/L。方法精密度在4—8％之间,回收率为90—102％。     

Biogeochemistry of Mangrove Soil Organic Matter: a Comparison BetweenRhizophoraandAvicenniaSoils in South-eastern Brazil

Soil core samples fromRhizophora mangleL. andAvicennia schauerianaStapf & Leech forests from south-eastern Brazil were analysed for their total organic matter content and their sugars, amino acid and amino sugars composition. Organic carbon and nitrogen contents were higher inAvicenniathan inRhizophorasoils. The contribution of sugars and amino acids to the total organic carbon pool was constant with depth inRhizophorasoils whereas inAvicenniasoils it increased. Spectral distribution of sugars and amino acids showed a dominance of Ca-affine monomers, particularly acidic amino acids, and the sugar arabinose. Biogeochemical indicators derived from ratios of individual sugar and amino acid monomers confirm previous studies which showed that organic matter in both soils is mainly of mangrove origin. The results further suggest accumulation of organic matter inRhizophorasoils and a continuing degradation of organic matter inAvicenniasoils. The latter may thus release more nutrients to adjacent ecosystems thanRhizophorasoils.     

A review of orange roughy Hoplostethus atlanticus fisheries,estimation methods,biology and stock structure

Orange roughy Hoplostethus atlanticus are unusual fish. They form dense aggregations that have fuelled lucrative fisheries at great depths (600–1 400 m), especially off Namibia, New Zealand and Australia. They are thought to be very long-lived (>100 years, maturity at 22 – 40 years), and to have exceptionally low natural mortality (M = 0.045–0.064 year^?1) and slow growth rates (K = 0.055–0.070 year^?1). In addition, they spawn large eggs and have low fecundity. These factors combine to make orange roughy highly susceptible to overfishing; most stocks are below 30% of pristine levels. Assessments are obtained from indices of catch rate and trawl, acoustic and egg surveys. Acoustic estimates are the most direct, but are confounded by the species' low target strength (?50 to ?53 dB)–attributable to the wax-filled swim bladder. Extracellular wax esters are stored in abundance and comprise mostly mono-unsaturated fatty acids, with low concentrations of the ω-3 fatty acid family. This unusual composition (resultant from the species' diet) ensures neutral buoyancy. Stock separation has been inferred mainly from biological studies, but genetic studies have also found differences among stocks within New Zealand and Australia. Deep-water habitat may be damaged by trawling operations and may take many years to recover, so in some quarters there is a call for a portion of suitable habitat to be set aside for preservation. Although Namibian orange roughy are shallower, smaller and younger than those in other stocks, the Namibian fishery sustained high catches for only a few years before quotas were reduced, from 12 000 to 1 875 tons. Three management lessons are suggested for developing orange roughy fisheries based on the Namibian experience: (1) imposition of catch limits during exploratory fishing; (2) starting the acoustic surveys earlier in the fishery, if possible; (3) greater reliance on trends in catch rate until a survey series has been established.