斑节对虾白斑综合症杆状病毒感染的实验模式

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。     

对虾一种无包涵体杆状病毒病原的PCR检测

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.     

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

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.     

Cloning of a chub metallothionein cDNA and development of competitive RT-PCR of chub metallothionein mRNA as a potential biomarker of heavy metal exposure

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.     

用16S rRNA基因的内切酶图谱快速鉴别几种对虾病原菌

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     

钙质超微化石定量研究方法

针对当前从事钙质超微化石定量研究使用的不同制样方法，简要介绍了适用于光学显微镜（LM）和扫描电子显微镜（SEM）的传统涂片法、小珠校准法、随机沉淀法和过滤法的主要流程和设备要求，同时对不同方法的优缺点进行了评述。     

Diagnosis of iridovirus in large yellow croaker by PCR

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.     

Seasonal Diatom Variability and Paleolimnological Inferences – A Case Study

The seasonality of physical, chemical, and biological water variables is a major characteristic of temperate, dimictic lakes. Yet, few investigations have considered the potential information that is encoded in seasonal dynamics with respect to the paleolimnological record. We used a one-year sequence of diatoms obtained from sediment traps and water samples, as well as the sedimentary diatom record covering the past ca. 1000 years in Bates Pond, Connecticut (USA), to investigate which variables influence the seasonal distribution of diatoms and how this can be used for the interpretation of the fossil record. The seasonal patterns in diatom assemblages were related to stratification and, to a lesser extent, to nitrate, silica, and phosphorus. During mixing periods in spring and autumn, both planktonic and benthic species were collected in the traps, while few lightly silicified, spindle-shaped planktonic diatoms dominated during thermal stratification in summer. Changes in fossil diatom assemblages reflected human activity in the watershed after European settlement and subsequent recovery in the 20th century. A long-term trend in diatom assemblage change initiated before European settlement was probably related to increased length of mixing periods during the Little Ice Age, indicated by the increase of taxa that presently grow during mixing periods and by application of a preliminary seasonal temperature model. We argue that the analysis of seasonal diatom dynamics in temperate lakes may provide important information for the refinement of paleolimnological interpretations. However, investigations of several lakes and years would be desirable in order to establish a more robust seasonal data set for the enhancement of paleolimnological interpretations.     

Prediction of volumes and risk in hydrocarbon exploration: a quantification of geology

 The prediction of the hydrocarbon potential of a specific trap or of a number of specific traps (venture), referred to herein as prospect appraisal, concerns a probabilistic exercise based on the quantification of geology in terms of structural closure, reservoir quality, hydrocarbon charge, and the retention potential of the seal. Its objectives include: (a) prediction of the hydrocarbon volumes that could be present in the trap from an analysis of its geologic attributes; (b) the amount of uncertainty introduced in the volumetric prediction by the uncertainties in the subsurface geology; (c) the risk that one or more of the essential attributes of the prospect are underdeveloped and recoverable reserves are absent. The uncertainty of the geologic input requires a probabilistic approach, for which the Monte Carlo procedure is well suited. Prospect appraisal forms the basis for decision-making in oil exploration and development and, therefore, should be reliable, consistent, and auditable. This requires the use of a consistent methodology, the development of reliable models to quantify the geologic processes involved, and the collection of comprehensive and relational databases for the many geologic variables. As a result of data availability, uncertainty and risk tend to increase strongly from mature, producing basins to areas of frontier exploration. This may complicate management of exploration portfolios. Received: 1 July 1996/Accepted: 25 November 1996     

基于最优分割分级法的月球撞击坑分级及其演化分析

撞击坑是月球表面广泛分布的重要构造形态,占据了月球表面的大部分面积。撞击坑的直径差别很大,从几微米到数百千米,其退化程度与形成年代具有密切关系。为了研究不同地质年代形成的撞击坑直径大小及其演化规律,需采用量化分级方法对大小不同的撞击坑进行定量分级和统计分析。本文在月表撞击坑数据库LU60645GT和Lunar_Impact_Crater_Database(2011)的基础上,结合数据库中撞击坑的直径、深度和年代信息,利用最优分割分级法对撞击坑直径进行定量化分级,并根据分级结果,综合分析撞击坑几何形态特征及其演化规律。研究结果表明,撞击坑形态特征的演化与年代有密切的关系。在相同级别、相同地体下,撞击坑形成的年代越早,其形态特征的精细结构退化程度越明显,只保留了大体的几何形状;而在不同级别、相同地体、相同年代下的撞击坑形态特征则由简单逐渐变为复杂,坑物质也逐渐变得复杂。