斑节对虾白斑综合症杆状病毒感染的实验模式     

邱德全  林继辉  刘丽  张凌飞 《广东海洋大学学报》2001,21(1):10-13

在建立斑节对虾实验室养殖模式的基础上，对野外采集的有白斑综合症病毒（WSSV）病典型症状的斑节对虾初步纯化其病毒，并进行WSSV的PCR检测，得到WSSV的感染用样品。对健康斑节对虾分别进行浸浴感染，投喂感染和注射感染。对感染死亡个体进行WSSA的PCR检测和细菌检测，证实WSSV感染性和致死性。浸浴感染、投喂感染和注射感染的感染量分别为4mL/L、0.2g/10g虾体、1/2稀释液0.05mL/10g虾体，死亡开始时间分别为16d、42h、28h，三种感染模式最终死亡率100%，从开始死亡到全部死亡延续时间分别为15d、82h、44h。  相似文献   

Quick-look assessments to identify optimal CO2 EOR storage sites   总被引：1，自引：0，他引：1  

Vanessa Núñez-López  Mark H. Holtz  Derek J. Wood  William A. Ambrose  Susan D. Hovorka 《Environmental Geology》2008,54(8):1695-1706

A newly developed, multistage quick-look methodology allows for the efficient screening of an unmanageably large number of reservoirs to generate a workable set of sites that closely match the requirements for optimal CO₂ enhanced oil recovery (EOR) storage. The objective of the study is to quickly identify miscible CO₂ EOR candidates in areas that contain thousands of reservoirs and to estimate additional oil recovery and sequestration capacities of selected top options through dimensionless modeling and reservoir characterization. Quick-look assessments indicate that the CO₂ EOR resource potential along the US Gulf Coast is 4.7 billion barrels, and CO₂ sequestration capacity is 2.6 billion metric tons. In the first stage, oil reservoirs are screened and ranked in terms of technical and practical feasibility for miscible CO₂ EOR. The second stage provides quick estimates of CO₂ EOR potential and sequestration capacities. In the third stage, a dimensionless group model is applied to a selected set of sites to improve the estimates of oil recovery and storage potential using appropriate inputs for rock and fluid properties, disregarding reservoir architecture and sweep design. The fourth stage validates and refines the results by simulating flow in a model that describes the internal architecture and fluid distribution in the reservoir. The stated approach both saves time and allows more resources to be applied to the best candidate sites.  相似文献   

概率图解法软件研制及其在东升庙矿床研究中的应用     

王琳  彭润民  韩雪峰  林美春 《地质与勘探》2008,44(1):89-93

文章根据概率图解法原理,利用VB可视化编程技术,完成了概率图筛分软件的研制.并通过该软件对东升庙矿床矿石品位混合总体进行筛分处理,探究其成矿期次信息.已有研究结果表明:一次成矿作用形成的矿床,其矿石品位的累积概率图是一条直线.而东升庙矿床的8条代表性勘探线中4种主要成矿元素TS、An、Pb、Cu品位的累积概率图均为曲线且出现明显拐点,经多次作图检验证实有效拐点数为1,由此得到新的成矿期次信息,揭示该矿床经历了两期成矿作用,即中元古代同沉积-海底喷气矿化期和后生改造矿化期,这对于研究其成矿物质来源和成矿机理具有重要的意义.  相似文献   

对虾一种无包涵体杆状病毒病原的PCR检测   总被引：4，自引：0，他引：4  

石拓  孔杰  刘萍  包振民  韩玲玲  徐怀恕 《海洋学报》2000,22(4):96-100

为建立对虾无包涵体杆状病毒的快速诊断技术,应用PCR技术分别对暴发性流行病发生前、流行中和发病后的对虾样品进行了检测.根据已克隆的对虾杆状病毒部分基因组序列而设计的引物能特异性地扩增出靶DNA片段,最低可以检测1pg的病素DNA.由典型发病症状对虾的胃、腮、肝胰腺、中肠、游泳足、肌内和心脏等器官和组织中成功地检测到病毒.不同组织和器官经扩增得到的信号强弱不同:病虾的胃扩增得到的信号最强,中肠、肌肉、心脏和游泳足次之,腮和肝胰腺信号最弱,说明病毒在不同组织和器官中数量及感染程度不同.在对虾发病前及发病后,用二次PCR还能检测表面不发病呈隐性感染状态的对虾携带的病毒;而对野生健康虾的检测结果为阴性.研究表明,PCR是检测对虾暴发性流行病快速、灵敏而准确的方法,对病毒病的早期诊断、防治和高健康对虾品种的选育具有指导意义.  相似文献   

Development of a real-time PCR method for Thalassiosira rotula rapid detection     

HE Shanying  YU Zhigang  MI Tiezhu 《海洋学报(英文版)》2007,26(2):133-139

Gene specific primers and DNA probe were designed based on the sequence of 18S rDNA cloned from the red tide alga Thalassiosira rotula. A real-time fluorescent quantitative PCR (RFQ-PCR) method was developed for quantitative detection of T.rotula. The RFQ-PCR assay data showed that the results obtained with the RFQ-PCR quite good agreement with those with the light microscope (LM) counting method, which suggested that the RFQ-PCR could be a useful method for red tide alga detection.  相似文献   

Cloning of a chub metallothionein cDNA and development of competitive RT-PCR of chub metallothionein mRNA as a potential biomarker of heavy metal exposure   总被引：3，自引：0，他引：3  

Hayes RA  Regondi S  Winter MJ  Butler PJ  Agradi E  Taylor EW  Kevin Chipman J 《Marine environmental research》2004,58(2-5):665-669

Metallothionein has been assayed in a range of aquatic animal tissues as an indicator of metal exposure. We sequenced chub (Leuciscus cephalus) metallothionein cDNA which showed over 90% homology to common carp, goldfish and stone loach and 77% homology to rainbow trout sequences for metallothionein. We then used the extended primer method to develop an accurate quantitative competitive RT-PCR assay for metallothionein mRNA. RT-PCR was used to measure metallothionein mRNA in feral chub from a range of field sites, with different levels of heavy metal pollution, in the West Midlands, UK. Measurements were complemented by analysis of liver and gill metallothionein protein by capillary electrophoresis. There was no significant difference in the metallothionein protein levels between fish of different rivers and there was no evidence of elevation of mRNA at the sites of highest metal exposure. The level of metal exposure (e.g. zinc, nickel and cadmium each ranging between 15 and 28 microg/l ) at the pH (7.5-8.5) of these rivers appears insufficient to elevate hepatic or gill metallothionein in chub. A lack of elevation of hepatic metallothionein mRNA in chub exposed to zinc, copper and manganese for 24 h and 10 days in the laboratory also suggests a non-responsiveness of this species.  相似文献   

用16S rRNA基因的内切酶图谱快速鉴别几种对虾病原菌   总被引：4，自引：0，他引：4  

孔杰  刘萍  张岩  杨丛海  叶军  徐怀恕  郭华荣 《海洋与湖沼》1997,28(5):449-452

坎普氏弧菌是青岛海洋大学生物系微生物实验室于１９８９－１９９０年自对虾养殖场中国对虾红腿病心脏及血淋巴中分离并鉴定的菌株，副溶血菌和溶藻胶弧菌两菌株于１９９４年９月得自中国科学院微生物研究所，为建立快速、准确的中国对虾病原菌的诊断技术，根据几种细菌的１６ＳｒＲＮＡ基因的序列，设计并合成该基因的多聚酶反应的引物ＰＬ１和ＰＬ２。并用该对引物分别从坎普氏弧菌、副溶血弧菌和溶藻胶弧菌的ＤＮＡ要品中扩增出分  相似文献   

Diagnosis of iridovirus in large yellow croaker by PCR   总被引：1，自引：0，他引：1  

Chen Xinhu  Wang Xiaowen  Wu Wenzhong 《海洋学报(英文版)》2003,22(4):635-642

A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus(LYCIV) is described,which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen.Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus(RSIV) and sea bass iridovirus(SBIV),suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone.The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes,the expected fragment was detected from spleen DNA samples of infected fishes,whereas no fragments were amplified from healthy fish spleen DNA,white spot syndrome baculoviruses(WSBV) DNA and pseudorabies virus(PRV) DNA.Detection limit of this method was 10^-7 ng positive plasmid DNA containing target sequence,equal to about 100 virions.In the infected experiment,first positive detection(1/4) appeared at Day 3 post-infection,all fish(4/4) tested positive at Day 7,however obvious symptoms were observed at Day 8,so LYCIV infection could be detected prior to the appearance of obvious symptoms.These results indicate that this PCR method could be used for early,rapid and specific detection of LYCIV infection.  相似文献   

东海原甲藻cDNA文库构建及尝试性EST分析   总被引：2，自引：0，他引：2  

张秀芳  杨官品  刘永建  李瑞香  朱明远 《中国海洋大学学报(自然科学版)》2006,36(3):361-364

东海原甲藻(Prorocentrumdonghaiense)是中国沿海频繁发生的大规模赤潮的原因种之一,大规模分离鉴定东海原甲藻功能基因是理解东海原甲藻赤潮形成过程和机理的重要基础。取对数生长期藻体,经微量总RNA抽提、cDNA合成、cDNA扩增和克隆等步骤,构建了东海原甲藻cDNA文库并进行了尝试性表达序列标签分析。从含有约5000个转化子的文库中随机取150个测序,获得126条EST序列。经网上BlastN及BlastX分析,共发现11个是已知功能的基因的标签。这些基因与东海原甲藻生长发育、物质转换和能量代谢相关。  相似文献   

我国陆上大中型气田分布规律与有利勘探区带优选     

曾宪斌  张静华  金惠  袁素华 《海洋地质与第四纪地质》2000,20(3):79-82

“八五”、“九五”期间我国天然气探明储量幅度增长,大中型气田分布规律的深化研究密切关系到天然气储量的增长势头。克拉通盆地下层序古隆起控气理论和上层序非构造圈闭控气机理是克拉通大中型气田富集分布的基本规律,前陆盆地是我国富气盆地类型之一,其冲断带下盘大型造圈闭群、前缘斜坡岩性尖灭区带与浅层次生气藏、前缘隆起上的断块圈闭群等是该类盆地有利的天然气富集区带。依据充分的地质资料,结合我国天然气勘探实际,提  相似文献