网箱养殖石斑鱼病毒性神经坏死病流行调查

厦门海水网箱养殖石斑鱼春、夏季节暴发性疾病经过流行病学调查和病原检测,确认主要病原为病毒性神经坏死病毒,病鱼外观表现典型的神经异常症状。应用逆转录聚合酶链式反应（RT—PCR）技术检测5种石斑鱼：赤点石斑鱼（Epinephelus akaara）、青石斑鱼（E．awoara）、云纹石斑鱼（Emoara）、紫石斑鱼（E．lanceolatus）、马拉巴石斑鱼（E．malabaricus）.检测76条石斑鱼,检出率90％;病鱼的脑、视网膜、肾脏、肝脏、脾脏等组织均能检出神经坏死病毒（NNV）,以脑和视网膜检出率最高,肾脏次之,肝脏、脾脏检出率较少,鳃未检出。     

白斑综合症病毒膜蛋白VP124参与病毒的感染

白斑综合症病毒(white spot syndrome virus,WSSV)是对虾的最主要病毒病原之一,给对虾养殖造成了巨大的经济损失。WSSV分别与三种该病毒膜蛋白VP39,VP124和VP187的特异性抗血清在体外作用后,再注射到螯虾体内,进行中和试验。结果显示,注射同或未同VP124抗血清作用的WSSV,8 d后螯虾的死亡率分别是100%和60%,WSSV的感染能够被抗VP124抗体中和。进一步利用定量PCR分析上述三种特异性抗血清对病毒感染的影响,结果显示,VP124抗血清可抑制WSSV在螯虾体内的增殖。所有结果表明,VP124在病毒的感染中起作用。     

Occurrence of gravel in the stomachs of two New Zealand stream‐dwelling fish (note)

Gravel occurred in. the stomachs of 17.8% of 924 Galaxias vulgaris and 2.8% of 430 Philypnodon breviceps taken from the Glentui River, Canterbury, between June 1970 and November 1971. It is suggested that differences in feeding habits and feeding locality are responsible for the difference in occurrence of gravel in the stomachs of the two fish species. Galaxias vulgaris feeds in the water column in riffles and, being a drift feeder, is more likely to ingest inedible material by accident than is P. breviceps, which feeds on the bottom in sheltered or quiet stretches of the river.     

The application of GIS for flood defence in the Anglian Region: developing for the future

The spatio‐temporal relationship between unusual sightings of dead birds and human West Nile virus infections has been observed in many studies and has been proposed as an indicator of an intense amplification cycle between birds and mosquitoes. However, to date, no single study has provided quantitative evidence that the amplification cycle occurs at the local level and that it operates within certain temporal parameters. Here, we use a novel geostatistical and spatial analytic methodology and present the first evidence that the localized unusual space–time correspondence of dead birds models the amplification cycle and that this cycle peaks 15–16 days prior to human onset of West Nile virus infections. During the process of establishing this relationship, we extend the traditional Knox space–time interaction measure to overcome pair‐dependency limitations and use a novel implementation of the kappa non‐chance agreement measure to identify the temporal characteristics of the association of bird deaths to human West Nile infections.     

Reduced removal of bacteriophage MS2 in during basin infiltration managed aquifer recharge as basin sand is exposed to infiltration water

Basin infiltration managed aquifer recharge (MAR) is a commonly used method for storing and treating surface water to be used as drinking water. This study examined how the removal of bacteriophage MS2 was affected by the relative age of the sand used for basin infiltration MAR at 4 °C using batch experiments (static and agitated) and column experiments. The sand and the water used in all experiments were characteristic of that used at a basin infiltration MAR scheme in Uppsala, Sweden. Experimental data was fit with numerical models describing the fate and transport of virus in soil—water systems. The “used” sand that had been subjected to intermittent infiltration over a period of 8 years had 15 times the amount of organic carbon than the “new” sand, which had yet to be used for infiltration. Results showed that attachment of MS2 to the new sand in batch experiments was relatively irreversible. Attachment to the used sand was reversible. Inactivation of MS2 was slowed when it was attached to the used sand. Results for the column experiments showed that the removal rate of MS2 was significantly lower in columns of used sand than in columns of new sand. Simulations indicated that MS2 would be entirely removed in the infiltration basins with new sand. Less than 3 log removals was estimated for basins with used sand. Reduced removal of MS2 by the used sand was deemed to be most likely due to organic coatings on the used sand. Results of this study give deeper insight into the mechanisms responsible for removing virus in infiltration basins and how those mechanisms will change as the sand in the basin is exposed to infiltration water.     

White spot syndrome virus inactivation study by using gamma irradiation

The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus(WSSV). White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3–10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 10 5.4 /mL. Shrimp post-larvae(1–2 g) were treated with gamma-irradiated(different doses) WSSV(10 0 to 10-4 dilutions) for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14–15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gammairradiated WSSV especially at 14–15 KGy.     

Effects of virus infection on expression of cell cycle regulatory proteins in the unicellular marine algae Emiliania huxleyi

The authors have investigated the biochemical events by which marine algal virus infection induces cell cycle arrest. The key G₂/M-phase regulatory proteins are analyzed by immunobloting in unicellular Emiliania huxleyi, suggesting that virus induced cell cycle arrest is related with virus''s effect on cyclins and cyclin dependent kinases. E. huxleyi virus (EhV) represses Cdc2/cyclinB complex activity by inhibiting the activity of Cdc2 kinase in a phosphorylation-related manner, blocking host cells G₂/M checkpoint. Dephosphorylated/inactive Cdc25C combined with up-regulation of Wee1 expression at early infect period appears to be important mechanisms by which EhV represses Cdc2/cyclinB complex activity that is required for entry into M phase. This study has allowed us to confirm that algal virus infection leads to selective activation or inhibition of certain cell-cycle factors, which may play a significant role in establishing a more efficient environment for viral gene expression and DNA replication.     

猪传染性胃肠炎病毒S、N基因核酸疫苗的构建与免疫原性试验

采用RT—PCR扩增TGEVSCH株S基因5’端主要抗原位点编码区和全长N基因,将其插入真核表达载体pVAXl,构建单独与嵌合表达S、N基因的pVAX—S、pVAX—N、pVAX—S—N3种重组质粒。通过脂质体将重组质粒体外转染COS-7细胞,利用间接免疫荧光检测目的基因的表达情况。将6周龄NIH小鼠随机分为5组,分别于腿部肌肉注射重组质粒pVAX、pVAX—S、pVAXS-N、pVAX—S＋pVAX—N与pVAX—S＋pVAX,共免疫3次,间隔2周。通过间接EI。IsA以及流式细胞仪分别检测免疫小鼠血清中特异性TGEVIgG抗体和外周血T淋巴细胞亚群数量。结果显示,重组质粒构建正确且在COS-7细胞中得以表达;pVAX—S＋pVAX—N质粒混合免疫组与pVAX—s＋pVAx质粒混合免疫对照组相比,可有效提高免疫小鼠外周血T淋巴细胞亚群数量和血清中特异性抗体水平的增加,而pVAX—S-N免疫组与pVAX—S组比较,未能起到提高免疫小鼠血清中特异性抗体水平的和外周血T淋巴细胞亚群数量作用。以上结果表明S、N基因的联合应用可有效加强TGEV核酸疫苗的免疫效力,但这种增强作用与S、N基因联合应用的方式直接相关。     

口蹄疫病毒VP1植物表达载体的构建

P1 T重组质粒上含有口蹄疫病毒 (FMDV)GD10分离株的p1cDNA片段 ,以此为模板 ,用PCR方法扩增其中的VP1基因 ,获得大小约 6 40bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体 pCAMBIA130 5 .2 ,转化EcoliTOP10感受态细胞。重组质粒经PCR、酶切及序列分析 ,证实VP1基因处于CaMV35S启动子控制 ,且读码框正确