一种提取饼形碘泡虫的简易方法

<正> 饼形碘泡虫主要寄生在草鱼的肠组织中,其大小只有几微米,碘泡虫的多数孢子由肠组织分泌的结缔组织包围形成孢囊,比较难分离和提纯。在研究该孢子虫的形态特征、生物学特性及防治方法等试验中,必须备有大量纯净的孢子.为了满足上述的需要,作者试验得出一种简单易行的方法。此种方法不需任何     

装甲RNA在牡蛎中诺如病毒4种RNA提取方法比较研究中的应用

RNA提取是诺如病毒检测的关键步骤,而目前贝类中诺如病毒RNA提取、检测方法的比较与评价常囿于缺乏量值明确、无生物安全隐患的标准样品作为参考依据。本研究将前期制备的 GⅡ型诺如病毒装甲RNA (3.0×1010拷贝)作为标准样品,人工污染牡蛎(Ostrea gigas tnunb)消化腺匀浆物,用4种常见RNA提取方法：TRIzol试剂、Viral RNA Kit、High Pure Viral Nucleic Acid Kit、柱式病毒RNAOUT试剂盒,分别提取RNA,经实时荧光RT-PCR检测后,利用标准曲线进行定量分析,分别计算4种方法对装甲RNA的回收率。结果显示,对于匀浆样本,TRIzol法对装甲RNA的回收率最高(6.80±0.89)%,显著高于Viral RNA Kit (4.51±2.28)%,二者的回收率又显著高于High Pure Viral Nucleic Acid Kit (0.24±0.05)%与柱式病毒RNAOUT试剂盒(0.11±0.02)% (P?0.05);对于冻干样本,Viral RNA Kit对装甲RNA的回收率最高(8.71±0.17)%,显著高于TRIzol试剂(7.12±0.64)%,二者的回收率又显著高于High Pure Viral Nucleic Acid Kit (0.33±0.12)%与柱式病毒RNAOUT试剂盒(0.06±0.01)% (P?0.05)。研究表明,TRIzol试剂与Viral RNA Kit对牡蛎消化腺样本中人工添加的装甲RNA均有良好的回收效果,同时也提示装甲RNA可作为一种良好的标准样品用于不同RNA提取试剂盒方法的评价与比较研究。     

多重RT-PCR体系检测4种虾病毒的方法

根据多重RT-PCR的技术原理,利用对虾传染性表皮与造血组织坏死症病毒、白斑综合征病毒、黄头病毒和桃拉综合征病毒的基因序列分别设计了4对特异引物,建立多重RT-PCR体系用于虾4种病毒的检测。多重RT-PCR体系能特异地扩增出IHHNV、WSSV、YHV和TSV的目的片段:TSV特异性扩增片段508 bp,WSSV 特异性扩增片段435 bp,IHHNV 特异性扩增片段301 bp 和YHV。特异性扩增片段614 bp。结果表明,多重PCR虾病毒检测系统具有较高的特异性和敏感性,并对其它对虾病原呈阴性。IHHNV、TSV、WSSV和YHV模板在多重PCR虾病毒检测体系中的检测下限分别为0.1,1,0.02和0.2 pg。病毒感染病料检测试验中,该检测体系的检测结果与单纯PCR的检测结果呈现出较好的吻合度。     

中华绒螯蟹1种球状病毒颗粒的电镜观察

中华绒螯蟹(Ｅｒｉｏｃｈｅｉｒｓｉｎｅｎｓｉｓ)养殖病害问题已成为制约养殖生产的主要因素,病害造成的损失逐年增长,损失也越来越严重。1996年以来,江苏、浙江、安徽等地相继发生养殖中华绒螯蟹患颤抖病成批死亡的病例,养殖单位采用多种抗菌药物治疗均无明显效果。病蟹...     

暗纹东方鱼屯的一种病毒性疾病

发病暗纹东方鱼屯以体表和内脏出血为主要症状,死亡率可达80%以上。显微镜检查体表、鳃丝及肠腔未见寄生虫。病鱼的肝脏和血液中未能分离到病菌。经电镜检查,在肝脏组织中发现大量球形或多角形病毒颗粒,核衣壳直径约85 nm,双层衣壳,病毒核心直径50~60 nm,无囊膜,未见包涵体。病毒颗粒常聚集成团,呈晶格状排列,聚集团外有包膜,有的病毒颗粒分散于细胞中。病毒的形态结构及大小与呼肠孤病毒相似。对该病的防治方法作了讨论。     

暗纹东方鲀的一种病毒性疾病

发病暗纹东方鲀以体表和内脏出血为主要症状,死亡率可达80%以上.显微镜检查体表、鳃丝及肠腔未见寄生虫.病鱼的肝脏和血液中未能分离到病菌.经电镜检查,在肝脏组织中发现大量球形或多角形病毒颗粒,核衣壳直径约85 nm,双层衣壳,病毒核心直径50～60 nm,无囊膜,未见包涵体.病毒颗粒常聚集成团,呈晶格状排列,聚集团外有包膜,有的病毒颗粒分散于细胞中.病毒的形态结构及大小与呼肠孤病毒相似.对该病的防治方法作了讨论.     

刺参体内的新病原--一种球状病毒

2004年12月中下旬~2005年4月对辽宁沿海地区刺参越冬苗发病严重地区,进行了跟踪调查分析。调查结果显示,2004年刺参越冬苗发病比往年早,来势凶猛,持续时间长,流行面广,具较明显的流行病特征,病症异于往年,发病厂家达50%以上,部分地区超过80%,严重地区80%~90%;从患病参体内分离出2株优势细菌和一种新的球状病毒,经负染和超薄切片在电镜下观察,证实有大量球状病毒存于参体内。     

同时检测两种对虾病毒和4种弧菌的同步PCR方法的建立

通过检索、多重比对、分析和筛选GenBank中对虾白斑综合征病毒(WSSV)、传染性皮下和造血器官坏死病毒(IHHNV)、副溶血孤菌、创伤弧菌、哈维氏弧菌和溶藻胶弧菌的基因序列,设计了10对特异性引物,以已知毒株和菌株的DNA为模板进行PCR,均能扩增出与实验设计相符合的DNA片段,对PCR扩增条件进行优化,建立了可同时检测鉴别WSSV、IHHNV、副溶血弧菌、创伤弧菌、哈维氏弧菌和溶藻胶弧菌,并且能同时区分WSSV不同地理毒株的同步PCR方法.研究结果表明,该方法检测特异性好,检测通量大,适合于对虾多种病原的同时检测.     

水产品中诺如病毒富集方法的优化研究

针对检测过程中影响病毒富集的主要因素,按正交试验设计了9种不同的病毒富集方法,利用RT-PCR对富集方法进行检测和分析评价。结果表明,吸附缓冲液pH对NVs富集有显著影响（P〈0.05）,吸附剂、沉淀剂种类对富集作用无显著差异（P〉0.05）。在吸附缓冲液中性环境（pH7.5）下,以苏氨酸作为吸附剂吸附两次,用PEG6000与PEG8000混合沉淀两次为最优富集方法,病毒回收率为23%,检测灵敏度为150pgRNA。利用此法对青岛地区贝类样品进行了检测,阳性检出率为12.5%,表明优化的富集方法可以满足实际检测的要求。     

“红肉病”文蛤中发现的一种球形病毒的形态发生与细胞病理学

在患“红肉病”文蛤的消化盲囊上皮细胞中发现了一种球形病毒及其发生基质。病毒粒子呈球形，无囊膜，直径约50－80nm，在细胞质内形成多泡状的包涵体并进行病毒粒子的装配。受感染的细胞具有明显的病理变化，主要表现为核固缩或肿胀并出现核空泡，内质网膨胀、大部分转化泡状，线粒体溶解或固缩，溶酶体数量增多等。     

A simple method for estimating MSY from catch and resilience

The Law of the Sea requires that fish stocks are maintained at levels that can produce the maximum sustainable yield (MSY). However, for most fish stocks, no estimates of MSY are currently available. Here, we present a new method for estimating MSY from catch data, resilience of the respective species, and simple assumptions about relative stock sizes at the first and final year of the catch data time series. We compare our results with 146 MSY estimates derived from full stock assessments and find excellent agreement. We present principles for fisheries management of data‐poor stocks, based only on information about catches and MSY.     

ONPG单管定量检测法与国标法在检测海水中大肠菌群的比较研究

本实验室根据ONPG显色技术原理建立了大肠菌群单管定量检测技术,即在此基础上利用ONPG单管定量检测方法和国标法对实验室制备的标准菌的菌悬液和水样进行检测,对2种方法的检测结果进行比对.研究结果显示:两种方法对菌悬液、水样的检测结果无统计学差异,ONPG单管定量检测方法检测数据的准确性、精确性和重现性都高于国标法;ON...     

一种简易实用的活鱼运输方法——巨型尼龙袋充氧运输活鱼初探

巨型尼龙袋与常规尼龙袋相比,因其型号巨大而得名。巨型尼龙袋充氧运输活鱼是一个创新,具有操作简单、管理方便、装载量大、成活率高等优点．是一种经济实惠的运输方法。 一、国型尼龙袋充氧运输活鱼的基本原理 活鱼运输的关键．一是要求水质清新;二是要求氧气充足。巨型尼龙袋充氧运输活鱼,既能随时更换新水,又能及时补充氧气,因而装载量大,成活率高。 二、巨型尼龙袋的主要结构及操作 １．材料和结构。巨型袋采用无色透明的聚乙烯塑料薄膜做材料．每个袋长２ｍ～３ｍ．宽１ｍ～２ｍ．每袋配备一根橡胶管,管长１．５ｍ左右。为安全…     

WSSV和IHHNV对虾病毒的胶体金免疫层析快速检测

为了快速、方便地检测出这2种病毒,研究将PCR核酸扩增的高灵敏度和胶体金免疫层析技术快速、简便、直观的优点结合起来,建立了同时检测2种对虾病毒(WSSV和IHHNV)的同步PCR-胶体金免疫层析检测方法。采用了生物素标记和地高辛(Digoxigenin)标记引物。PCR扩增产物利用胶体金免疫层析技术进行检测。胶体金免疫检测试剂条的检测线(T线)处的层析膜处点上地高辛的单抗,经地高辛标记的PCR扩增产物可在T线上被检测到,在10 min内即可得到检测结果。同时,在PCR过程中使用UNG(尿嘧啶-DNA糖基酶),极大地控制了遗留污染。通过此方法检测对虾WSSV和IHHNV病毒的灵敏度均可达到10~100个拷贝,高于国家标准PCR检测法10~100倍。此检测方法让工作人员避免了接触有毒和诱导突变的试剂,免除基层检测实验室购买电泳设备和荧光检测系统的投资。     

Comparative assessment of eDNA metabarcoding and longline deployments for elasmobranch surveying across a large tropical marine park network

相似文献   

鳗鱼及其制品中喹诺酮类药物残留的微生物快速检测方法研究

介绍了一种采用E.coliATCC8739为检测菌,检测鳗鱼中喹诺酮类药物残留的微生物抑制试验方法,该方法快速、简便,适用于大批量的样品检测。该方法的检测低限为环丙沙星20μg/kg;恩诺沙星25μg/kg;其它喹诺酮类药物50～250μg/kg,6～8h即出结果。鳗鱼及其制品的恩诺沙星的回收率为75.89%～83.12%。     

A simple and rapid method for the analysis of fish histamine by paper electrophoresis

ABSTRACT:   The described analytical method for histamine determination in fish and seafood consists of sample extraction, adsorption onto a paper disc, application of the paper disc onto electrophoresis paper, electrophoresis for only 10 min, drying, and color developing by Pauly's reagent. Histamine can be satisfactorily detected and completely separated from histidine, carnosine and other Pauly reagent-positive compounds. This method does not require expensive instrumentation and any tedious pretreatment to eliminate potential interference by other imidazole compounds, such as histidine or carnosine. This method can be used to detect histamine in multiple fish and seafood samples simultaneously that contain as little as 15 p.p.m. histamine (1.5 mg/100 g).     

Isolation and characterization of a rhabdovirus from co-infection of two viruses in mandarin fish

Co-infection of two viruses has been observed in mandarin fish (Siniperca chuatsi), but the two viruses have not been characterized. In this study, a rhabdovirus has been isolated from the co-infected two viruses extracted from the diseased mandarin fish, and its morphological structure and partial biochemical and biophysical characteristics have been observed and analyzed. The isolated rhabdovirus has a typical bullet shape, and is therefore called S. chuatsi rhabdovirus (SCRV). And, the isolated rhabdovirus produced a higher titer (10^8.5 TCID₅₀ ml^− 1) than did the co-infecting viruses (10^6.5 TCID₅₀ ml^− 1). Subsequently, the viral genome RNA was extracted, and used as template to clone the complete nucleoprotein (N) gene by RT-PCR amplification. Cloning and sequencing of the SCRV N protein revealed 42%-31% amino acid identities to that of trout rhabdovirus 903/87 and the rhabdoviruses in genus Vesiculovirus. SDS-PAGE separation of the isolated SCRV and other two rhabdoviruses also revealed obvious polypeptide profile difference. Moreover, the anti-SCRV N protein antibody was prepared, and the anti-SCRV N protein antibody only could recognize the SCRV N protein, whereas no antigenicity was detected in other two rhabdoviruses. The data suggested that the SCRV should be a rhabdovirus member related to the genus Vesiculovirus in the Rhabdoviridae.     

Development and application of fluorescence in situ hybridization (FISH) method for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella in cultured and natural seawater

ABSTRACT:   The toxic dinoflagellates Alexandrium tamarense (Lebour) Balech and A. catenella (Whedon and Kofoid) Balech produce potent neurotoxins, such as saxitoxin and gonyautoxin and have been mainly responsible for paralytic shellfish poisoning (PSP) in Japan. To prevent a negative effect on the fishery industry, it is necessary to identify these toxic species precisely and rapidly before and during the bloom. In this paper, a rapid and simple protocol of a fluorescence in situ hybridization (FISH) method using ribosomal RNA (rRNA)-targeted probes has been established for identifying the cultured strains and natural cells of A. tamarense and A. catenella . Using the FISH method established in this study, it was possible to identify these toxic species species-specifically and rapidly, within 30 min. The procedure of detection constituted three steps: (i) fixation/dehydration; (ii) hybridization; and (iii) washing; this made the identification simple. Moreover, this method did not require either special techniques or equipment, and the cost for detection was low. The specificity, rapidity, and simplicity of the developed method suggest that it might be useful for routine monitoring of these toxic microalgae.