Sequence analysis and quantitative detection of Norwalk-like viruses in cultured oysters of China

We isolated 4 Norwalk-like viruses (NLVs) contaminated oysters from 33 Chinese oysters collected from local commer-cial sources of Shandong Province. After amplification of the RNA-dependent RNA polymerase (RdRp) region of NLVs genomes with RT-PCR, the open reading frame 1 (ORF1) of the RdRp was sequenced and subjected to multiple-sequence alignment. The re-suits showed that NLVs in the four isolates belong to genogroup Ⅱ. The sequence comparison showed that the similarity between four Chinese oyster isolates were higher than 99.0%, which indicated that NLVs prevalent in close areas have high homogeneity in genome sequences. In addition, the most conserved sequences between diverse NLVs were used to design primers and TaqMan probes, then the real-time quantitative PCR assay was performed. According to the standard curve of GII NLVs, the original amounts (copies) of NLVs in positive patient's fecal isolate, positive Japanese oyster isolate, and the Chinese oyster isolate were 8.9×108, 1.25×108 and 4.7×101 respectively. The detecting limit of NLVs was 1×101 copies. This study will be helpful for routine diagnosis of NLVs pathogens in foods and thus for avoiding food poisoning in the future.     

当地球发起攻击……

四川汶川大地震震惊世界.笔者参阅美国<大众科学>数据,并结合中国、湖南某些地质灾害情况,予以介绍,以飨读者.海啸、火山爆发、地震、山体滑坡--这些大自然杀手的唯一确定的特性是,它们总会卷土重来,一而再,再而三.我们最有效的防御措施是:更有效的预测和保护.     

浅议气象信息网络保障和维护

分析了现代气象信息网络现状和气象局域网常见故障,归纳了一般性解决方法和应对措施,提出了气象网络维护的一些办法。     

中国对虾（Penaeuschinensis）杆状病毒的装配和包涵体形成过程研究

对中国对虾肝胰腺上皮细胞的透射电镜观察，发现杆状病毒可在对虾肝胰腺上皮细胞的细胞质内繁殖。增殖过程中在病毒发生基质表面及附近形成晶格状排列的包涵体，通过包涵体的形成、分化等过程，完成杆状病毒的装配。杆状病毒的装配过程似可分为５个时期：１．病毒发生基质形成期；２．未成熟包涵体期；３．成熟包涵体期；４．包涵体分化与病毒粒子装配期；５．侵染期     

免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度

利用免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度.以牙鲆鳃细胞系(FG)作为感染细胞,将生长旺盛的FG细胞接种于48孔培养板中培养至形成细胞单层,用2倍连续稀释的LCDV粗提液分别接种FG细胞.固定各稀释度LCDV感染后的FG细胞,孵育抗牙鲆LCDV单克隆抗体,其后再运用生物素-亲合素反应系统,以碱性磷酸酶底物APRed试剂盒发色.倒置显微镜观察,被病毒感染的FG细胞的细胞质呈现红色,未被感染细胞的细胞质呈无色.记录各稀释度病毒感染的阳性细胞孔数,按Reed-Muench法计算组织细胞培养半数感染量(TCID0).结果显示,免疫细胞化学法测得LCDV在FG的滴度为1.77×210 TCID50/mL.该法可以用来有效测定LCDV滴度,且结果直观、准确性较好,灵敏度较高.     

养殖杂色鲍暴发病超微病理学研究

从1999年至2002年在中国南方沿海许多养殖场的杂色鲍(Haliotis diversicolor Reeve)出现了严重的疾病,该病可感染各种规格的鲍,死亡很快,死亡率高。病鲍的外观症状表现为外套膜收缩,腹足表面变黑,肌肉变得僵硬。通过电镜观察,在肝胰腺、外套膜、肾、鳃及肠等组织中发现了一种球状病毒。病毒粒子大小为100～130nm。该病毒一般存在于间质细胞的细胞质中,为双层质膜所包裹。染病鲍细胞器如滑面内质网扩张、线粒体崩解,糖原粒、核糖体减少,核膜松疏、溶解、消失,核质边缘化。病毒的病原性通过对健康鲍的感染试验得到证实。     

White spot syndrome virus envelope protein VP124 involved in the virus infection

White spot syndrome virus （WSSV） is one of the major shrimp pathogens causing large economic losses to shrimp farming. In an attempt to identify the envelope proteins involved in the virus infection, purified WSSV virions were mixed with three antisera against WSSV envelope proteins （VP39, VP124 and VP187 ）, individually. And then they were injected intramuscularly into crayfish （Procambarus clarkii） to conduct in vivo neutralization assays. The results showed that for groups injected with virions only and groups injected with the mixture of virions and antiserum against VP124, the crayfish mortalities were 100% and 60% on the 8th day postinfection, individually. The virus infection could be delayed or neutralized by antibody against the envelope protein VP124. Quantitative PCR was used to further investigate the influence of three antisera described above on the virus infection. The results showed that the antiserum against VP124 could restrain the propagation of WSSV in crayfish. All of the results suggested that the viral envelope protein VP124 played a role in WSSV infection.     

对虾白斑综合症病毒透射电镜分析中病毒固定条件的优化

电镜观察是研究对虾白斑综合症病毒(WSSV)形态和结构的重要手段.本研究采用不同浓度的多聚甲醛和戊二醛对纯化的WSSV粒子进行固定,从中寻找最优的固定条件用于病毒的透射电镜(TEM)分析.实验结果显示,经过多聚甲醛或者戊二醛固定的WSSV粒子对吸附、染色等实验操作的耐受性较未固定样品有显著提高,在制样过程中没有发生明显的结构破坏.其中1.00％ (V/V)多聚甲醛常温固定20 min对WSSV粒子的结构保持得最好.进一步地分析表明,1.00％多聚甲醛固定后的病毒粒子更有利于金标免疫电镜的研究.     

三亚海域浮游病毒与细菌丰度的时空变化规律以及相关环境因子*

在三亚海域设置3个站位, 分别于各站位连续采集12个月表层水样。利用流式细胞仪进行浮游病毒及浮游细菌丰度的测定, 并对两者及其与环境因子之间的相关性进行研究, 同时对不同站位之间进行差异性分析。结果表明, 调查海域浮游病毒丰度(平均7.63×10⁶viruses·mL^-1)高于浮游细菌丰度(平均1.52×10⁶cells·mL^-1)。浮游病毒及浮游细菌丰度在三亚河口最高, 且有明显的季节变化, 冬春季高于夏秋季, 并且其与鹿回头半岛西侧、小东海之间均有极显著差异(P<0.01), 不同类群病毒代表的宿主类群也有所不同。调查海域总体水平浮游病毒丰度与浮游细菌丰度显著正相关(r=0.800, P<0.01); 叶绿素(Chl a)和氮盐(NO- 2、NO- 2+NO- 3、NH+ 4)是影响两者的关键因子。     

渤海海域浮游病毒和浮游细菌的丰度变化与环境因子之间的耦联关系

Uncovering the role of environmental factors and finding critical factors which harbor significant fractions in governing microbial communities remain key questions in coastal marine systems. To detect the interactions between environmental factors and distributions of virio-and bacterioplankton in trophic coastal areas, we used flow cytometry to investigate the abundance of virio-and bacterioplankton covering 31 stations in the Bohai Sea of China. Our results suggested that the average abundance of total virus(TV) in winter(～2.29×10~8 particles/mL)was slightly lower than in summer(～3.83×10~8 particles/mL). The mean total bacterial abundance(TB) was much lower in winter(～2.54×107 particles/mL) than in summer(～5.43×10~7 particles/mL). Correlation analysis via redundancy analysis(RDA) and network analysis among virioplankton, bacterioplankton and environmental factors revealed that the abundances of viral and bacterial subpopulations depend on environmental factors. In winter, only temperature significantly influenced the abundances of virio-and bacterioplankton. In summer, in addition to temperature, both salinity and nutrient(SiO_2) had a remarkable impact on the distribution of virioand bacterioplankton. Our results showed a clear seasonal and trophic pattern throughout the whole water system, which revealed that temperature and eutrophication may play crucial roles in microbial distribution pattern.