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     

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.     

基于EMD与神经网络的机械故障诊断技术

经验模式分解 (EMD)是分析非线性、非平稳信号的有力工具 ,它将信号分解为突出了原信号的不同时间尺度的局部特征信息的内在模函数 (IMF)分量。本文通过将各 IMF分量输入到 BP网络中进行训练学习和故障诊断 ,比直接输入原信号可以提高 BP网络对故障诊断的准确率 ,而且减少了训练时间。     

苏里格气田盒8气藏气水层测井识别研究

鄂尔多斯盆地苏里格气田盒8气藏为弹性驱动的河流相低效气藏,储层非均质性强,在开发中普遍见水。根据试采情况将气藏储层分为气层、水层、气水层三类,开展气、水层的测井识别研究。以区内26口井50个射孔层段各层的试油结果作为依据,选出了700个样本点作为判识建模的标本,以声波时差和中子孔隙度等8个原始测井值作为判识变量组合,采用模糊判别分析,建立了各层的判识模型。回判结果表明,模型对气层精度达到97％,水层为89％,气水层为94％。利用未参加建模的出水井射孔段对模型进行验证,判识结果基本符合实际情况,模型具有较高精度。     

波包传播诊断的理论基础和计算方法

文中提出了一种新的利用实际观测资料研究波包传播的诊断方法 (WPD) ,并通过理想资料和实际资料的计算证明了该方法的可用性和广泛性     

北疆夏季两次冰雹天气过程的对比分析

北疆夏季两次冰雹天气过程的对比分析发现，充足的水汽、足够大的上升运动和不稳定的层结果冰雹天气出现的必备条件，而降雹的强弱则与对流云图的发展和强度有关。