饲料中添加河豚毒素(Tetraodotoxine,TTX) 的河豚养殖

笔者试图通过对养殖红鳍东方过程中投喂添加河豚毒素的饲料提高其免疫力,降低鱼病发生率,还其自然野生状态下的本来面目,从而达到全面的健康化养殖。译出此文,供有关专业人员参考,并望有所启迪。     

酶联免疫吸附法在水产养殖中的应用

简要介绍了酶联免疫吸附法,并且就其在水产养殖中的应用进行了较详细的评述,主要包括ELISA用于水产品安全检测、水产动物病原菌检测和水产养殖其它领域的应用,旨在能促进该技术在水产养殖领域中的发展。     

酶联免疫法(EIA)测定鱼虾体内的氯霉素残留

氯霉素是一种高效、广谱的抗生素。最近几年残留在动物食品中的氯霉素对人造成的伤害引起人们的重视。一些发达国家如欧共体,美国已相继禁止或严格限制使用氯霉素。通过对几种检测方法的比较,EIA是一种快迅、准确方法,适用于批量检测。在此,笔者就鱼虾中氯霉素残留EIA检测做一概要和总结。     

酶联免疫(ELISA)法测定水产品中呋喃唑酮代谢物AOZ的残留

采用MaxsignalTM AOZ快速检测试剂盒检测水产品中呋喃唑酮代谢物。通过对精密度、回收率和检测限等项目的测定,验证了方法的可行性。呋喃唑酮代谢物标准液在0．05ng／mL～4．5ng／mL范围内具有良好的线性,微孔板孔间变异系数为3．2％～7．7％。样品批内变异系数为5．2％～8．4％,批间变异系数为6．3％～8．9％,平均回收率为94．5％～98．4％。试剂盒的理论检测下限为0．05μg／Kg。该方法灵敏度高,重复性好,适用于水产品中呋喃唑酮代谢物残留的检测。     

酶联免疫法（ELISA）检测水产品中氯霉素残留量的影响因素及控制方法

利用酶联免疫法（ELISA）检测水产品中氯霉素的残留量,具有操作简单、灵敏度高和特异性强等优点。但由于ELISA检测过程中涉及样品采集、试剂准备、加样、温育、洗涤、显色、比色和结果判定与报告等诸多步骤,影响因素较多。通过详细分析各个步骤的影响因素及控制方法,以期使检测结果更加准确可靠。     

一种新的酶联免疫法检测水产品中的己烯雌酚

对罗非鱼和对虾样品加标回收试验表明,一种新的己烯雌酚酶联免疫检测试剂盒对水产品中己烯雌酚检测具有简单、快速的特点,检测回收率和精密度均可以满足水产品中己烯雌酚检测的需要,该快速检测方法尚未见国内有文献报道用于水产品中己烯雌酚的检测。     

酶联免疫方法(ELISA)快速检测水产品中药物残留假阳性原因浅析

基于抗原抗体反应基础上建立的酶联免疫分析法(ELISA),具有操作简单、快速、灵敏度高、特异性强等优点,是目前水产品中氯霉素残留筛选最常用的方法,但在实际操作过程中会发现有假阳性存在.为此,本文分析了ELISA方法检测水产品中氯霉素残留假阳性的原因.     

酶联免疫法快速检测水产品中硝基呋喃类代谢物、氯霉素及氟苯尼考

应用特异性强的间接酶联免疫法,同时检测4种硝基呋喃类代谢物、氯霉素(CAP)和氟苯尼考(FF),建立快速分析水产品中6种药物残留量的方法.样品用盐酸进行消化,再由乙酸乙酯提取,使用酶联免疫试剂盒进行检测.实验结果显示,6种分析物均在其线性范围内,线性系数均大于0.995.在加标浓度为0.1、0.5和1.5μg/kg的加...     

酶联免疫法与液相色谱-串联质谱法测定喹诺酮类药物残留效果比对

喹诺酮类抗菌药又称吡酮酸类或吡啶酮酸类,是一类较新的合成抗菌药,经40多年的发展,目前共开发出了4代喹诺酮类药物,共计50多种药物。喹诺酮类药物可用于治疗呼吸道感染、泌尿生殖系统感染、消化系统感染及其他类的各种感染,还可用于抗肿瘤和抗病毒。由于是人兽共用药,除了其毒副作用对人体的直接危害外,动物食品中低浓度的喹诺酮药物残留容易诱发病菌耐药性,影响临床治疗效果。     

Toxicity of cultured puffer fish and seasonal variations in China

To confirm the toxicity or the tetrodotoxin (TTX) content of cultured puffer fish in China, 565 specimens of cultured puffer fish (Takifugu rubripes, Takifugu fasciatus or Takifugu obscurus, Takifugu flavidus and Takifugu bimaculatus) were collected from five provinces from November 2007 to November 2008. The toxicity of the skin, muscle, liver and testis/ovary was examined using methods based on a mouse assay for TTX. The results indicated that the skin, muscle, liver and testis of specimens were ‘non‐toxic’ in both of the mouse assays [<10 mouse unit (MU) g^?1 or 2.2 μg g^?1] from the viewpoint of food hygiene. However, TTX was detected from an ovary sample of the cultured T. rubripes collected from the Shandong province in March 2008, which was 125.43 MU g^?1 tissue. The average TTX contents in the muscle, liver and skin of T. rubripes from Hebei and Shandong provinces were the highest in May or July (1.31, 0.90 and 0.62 MU g^?1, respectively, in Hebei, and 1.57, 0.32 and 0.16 MU g^?1, respectively, in Shandong) and the lowest in January (<0.1 MU g^?1 or 0.02 μg g^?1). Thus, non‐toxic cultured puffer fish could be produced successfully in mainland China.     

酶联免疫法测定水产品中呋喃唑酮代谢物AOZ的残留

运用酶联免疫法对水产品中呋喃唑酮代谢物残留测定的方法进行研究。结果表明,稀释倍数对回收率影响显著,而乙酸乙酯提取次数、光照条件和放置时间对测定结果无显著影响。向样品中分别添加0.60、2.00、6.00 μg·kg^-1 3个浓度水平的呋喃唑酮代谢物时,平均回收率分别为99.2%、96.0%和100.0%;批内变异系数为1.76%～12.57%,批间变异系数为5.43%～8.58%,最低检测限为0.3 μg·kg^-1。该方法灵敏度高,重复性较好, 适用于水产品中呋喃唑酮代谢物残留的筛选。     

罗非鱼源无乳链球菌的间接ELISA快速检测方法

以罗非鱼源无乳链球菌为抗原,免疫新西兰兔获得高效价的多克隆抗体;以此多克隆抗体作为一抗,羊抗兔Ig G-HRP作为酶标二抗,建立无乳链球菌的间接ELISA快速检测法。采用棋盘滴定法确定抗原与一抗的最佳工作浓度分别为106 CFU/m L和1∶10 000;酶标二抗的最适工作浓度为1∶1000。病原菌检测灵敏度为每孔103 CFU。该方法标准化后具有快速、灵敏等特性,与海豚链球菌等其他常见水产病原菌无交叉反应;阻断实验中的阻断率达72.02%;交叉反应和阻断实验的结果显示该方法具有较高的特异性。将该方法标准化后检测了44株2007—2013年分离的罗非鱼无乳链球菌,阳性检测率为100%;对人工感染后死亡的30尾罗非鱼的脑组织进行检测,阳性检测率为100%;对人工感染后未死亡的20尾罗非鱼的脑,肝和脾组织进行检测,脑的阳性检测率为0%,肝的阳性检测率为25%,脾的阳性检测率为50%;表明该技术不仅能够检测已发病的罗非鱼,而且能够检测无症状的带菌罗非鱼,该方法的建立有助于快速准确地诊断由无乳链球菌引起的罗非鱼链球菌病。     

Validation of a KHV antibody enzyme‐linked immunosorbent assay (ELISA)

Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti‐KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme‐linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.     

太湖微囊藻毒素在罗非鱼体内的累积及生物降解的初步研究

为了解蓝藻水华期间微囊藻毒素在罗非鱼体内的分布及累积传递过程,2008年6月至8月采集了高密度蓝藻池塘及太湖网箱内的鱼样及水样,用ELISA法对鱼样和水样进行微囊藻毒素MC-LR含量的检测。结果表明：池塘水体微囊藻毒素MC-LR含量变化范围在0.123～0.514ug/L间,MC-LR含量随着藻密度的下降而降低,对照组水体MC-LR浓度显著高于实验组MC-LR含量。池塘鱼体肌肉组织微囊藻毒素MC-LR累积含量在1.194～3.615ng/g间,肝脏组织微囊藻毒素MC-LR累积含量显著高于肌肉组织。将池塘与网箱罗非鱼转至无微囊藻水体中暂养,跟踪检测MC-LR含量变化,池塘和网箱鱼体肌肉组织微囊藻毒素MC-LR含量均低于人体每日可耐受摄入量,而肝脏组织藻毒素MC-LR含量则分别需要经过10～20天自然生物降解后降低至安全摄入量之下。并讨论了微囊藻毒素在鱼体内的组织分布与食物链中的累积传递。     

应用Dot_ELISA技术检测草鱼出血病病毒的研究

报道了斑点免疫吸附ＥＬＩＳＡ（Ｄｏｔ－ＥＬＩＳＡ）快速检测草鱼出血病病毒（ＧＣＨＶ）的方法以及对提纯病毒、染毒细胞内病毒和病鱼组织的检测结果，并对该方法的灵敏度与ＳＰＡ－ＣｏＡ和常规ＥＬＩＳＡ法进行了系统的比较。结果表明，Ｄｏｔ－ＥＬＩＳＡ检出ＧＣＨＶ的最低含量为３ｐｇ，该灵敏度比ＳＰＡ－ＣｏＡ和常规ＥＬＩＳＡ法分别提高１０倍和２０倍，而且快速易行，是目前检测ＧＣＨＶ最为有效的方法，并可在草鱼出血病临床诊断和病毒疫苗质量鉴定等方面得到应用。     

The purification and development of a quantitative enzyme linked immunosorbent assay (ELISA) for the measurement of vitellogenin in diploid and triploid brook trout (Salvelinus fontinalis)

Vitellogenin (VTG) was purified from the plasma of estradiol-17β (E₂) treated male brook trout (Salvelinus fontinalis; bt) using gel filtration and anion exchange chromatography. An antiserum to bt-VTG was raised in rabbits, then used to detect bt-VTG by Western blot analysis. Purified bt-VTG and its antibody were then used to develop an antibody-capture, competitive enzyme-linked immunosorbant assay (ELISA). Confirmation of the purified protein as bt-VTG was based on the characteristics of high molecular weight (∼562 kDa), dominant plasma protein in vitellogenic females and induction by exogenous E₂ treatment in males. The antiserum recognized a major 184 kDa polypeptide as well as minor bt-VTG degradation products (150–66 kDa) in purified bt-VTG and in plasma from vitellogenic females. Low levels of antibody cross reactivity were shown with plasma from nonvitellogenic females, control rabbit serum, and several other antisera known to not contain VTG. The ELISA had a minimum detection limit of 8 ng ml⁻¹ and intra- and inter-coefficients of variation less than 9% and 15%, respectively. The ELISA demonstrated parallel binding slopes among dilution curves of purified bt-VTG standard and plasma from diploid and triploid brook trout females. Using the ELISA, maximum plasma VTG levels of 93.5±33.6 mg ml⁻¹ were detected in vitellogenic diploid females, whereas only 0.18±0.15 mg ml⁻¹ were detected in triploid females of the same age (n=5 for each). Diploid and triploid males cohabitating with vitellogenic females showed measurable levels of plasma VTG during vitellogenesis in females (i.e., 0.17 and 0.06 mg ml⁻¹, respectively (n=1)). This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of dietary Europium complex and Europium(III) on cultured pearl colour in the pearl oyster Pinctada martensii

In the present study, through a transmission experiment, the effects of europium complex and Eu³⁺ on the pearl colour in the pearl oyster Pinctada martensii were evaluated. Europium complex and europium salt were respectively enriched by dietary green algae platymonas subcordiformi, and their toxicity to algae were analysed by the 96‐h acute toxic experiment. Then, the green algae enriched europium complex and europium salt were respectively fed pearl oysters for 2 months, and the surface microstructures of cultured pearls were presented with scanning electron microscopy (SEM) and atomic force microscope(AFM). The results show that toxicity of europium complex to algae is much lower than Eu³⁺ from lowest no observable effect concentration LNOEC values and 96h‐EC₅₀ values and two compounds can modify pearl structure and optimize pearl colour from SEM and AFM images. Taken together, our results, for the first time, show that rare earth complex can improve the pearl quality. This study provides an innovative approach for additional investigation of the influence of rare earth elements on pearls.     

我国五个青蛤地理群体遗传变异的RAPD分析

本文利用RAPD方法对我国浙江、江苏、辽宁、天津和福建沿海青蛤地理群体的遗传变异和遗传结构进行了分析。从60条随机引物中,筛选出其中12条扩增效果好、条带清晰的引物进行扩增。结果表明,青蛤遗传多样性较丰富,五个群体的多态度在0.249～0.307之间,平均为0.278,按递减依次为江苏>福建>辽宁>浙江>天津;经分子方差分析(AMOVA),10.69%的遗传变异来自于群体间,89.31%来自于群体内,遗传分化除辽宁与江苏、天津群体间不显著外,其它群体间分化显著;根据遗传距离,用UPGMA法进行聚类分析表明,浙江群体与江苏群体,辽宁群体与天津群体分别先聚在一起,最后与福建群体聚类;S11、S424、S228、S4254个引物可以区分这五个青蛤地理种群,作为群体特征标记。     

铜铸熟若蟹中麻痹性毒素成分的分离和鉴定

在分析了日本珊瑚扇蟹铜铸熟若蟹甲壳组织和内脏组织中毒性含量的基础上,对毒素提取液进行分离纯化,并通过液相色谱法对其毒素种类进行分析。毒性分析结果显示,毒蟹个体的含毒量在37 500~116 000 MU范围,远远超过成人中毒致死量(5 000 MU),但个体间的毒含量有较大差异;毒蟹个体的甲壳组织的比毒性(909~1 610 MU/g)总体高于内脏组织(111~576 MU/g)。Bio-Gel-P₂凝胶柱层析纯化结果显示,粗毒素提取液通过反复纯化可以达到纯化目的,毒素的比毒性从4.8 MU/mg上升到30 MU/mg左右;毒素进一步通过Bio-Rex 70离子交换柱层析的分离纯化结果显示,毒素中约有9%为膝沟藻毒素群(GTXs,麻痹性毒素中的一类)。高效液相色谱分析结果显示,膝沟藻毒素群的主要成分为GTX3和GTX2,并含有少量的dcGTX2和dcGTX3。