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.     

干旱区地下水脆弱性特征及评价方法探讨

干旱区地下水脆弱性是地下水系统本身固有的不稳定属性，是系统结构、功能状态在人类活动干扰及气候变化等自然因素作用下具有的敏感性、易变性和弹性的综合反映。可以以河川径流中冰雪融水比重、地表径流入渗占地下水补给比例、地下水补给强度、地表水的引用率等十项指标（IRRUDQELTS）进行定量评价。通过对塔里木盆地南缘地下水脆弱性评价，表明该区属地下水严重脆弱区，其中又以民丰县、皮山县为极端脆弱，结果与实际情况相符合，说明该评价指标体系具有一定的科学性。     

定量分析几种Q矢量

结合1991年7月5日20:00～6日20:00一次典型的江淮梅雨锋暴雨过程，从台站实际业务工作需要考虑，细致、具体地比较分析了850 hPa、700 hPa及500 hPa 3个层次的准地转 Q 矢量散度场、半地转 Q 矢量散度场、非地转 Q 矢量散度场及湿 Q 矢量散度场与相应时刻地面降水场对应关系的差异，同时还针对每一种 Q 矢量，将其在850 hPa、700 hPa及500 hPa 3个层次的散度场对同时刻降水场的反映能力进行了比较。在定量比较的基础上，得到了对4种 Q 矢量诊断特性的具体认识:（1）在整个梅雨锋暴雨过程中，3个层次的半地转 Q 矢量散度场及准地转 Q 矢量散度场对雨区的反映能力较小，而非地转 Q 矢量的散度场和湿 Q 矢量的散度场对雨区的反映能力明显较前二者大，尤其是湿 Q 矢量散度场在每个层次的诊断能力基本都大于相应层次的其它 Q 矢量的散度场。（2）对于每一种 Q 矢量而言，基本都是在700 hPa的散度场与雨区的对应关系好于各自在850 hPa和500 hPa的散度场，尤其是700 hPa湿 Q 矢量散度辐合场对同时刻梅雨锋暴雨的强度及落区都有很好的指示作用。最后，基于理论的角度对各 Q 矢量的诊断特性进行了较为深入地探讨和比较分析，明确地指出了4种 Q 矢量存在理论前提上的差异。