冰川表面冰尘洞内病毒研究进展

病毒在驱动生态系统高生物多样性形成以及生态系统营养物和有机物的循环过程中均发挥着重要的作用。冰川洞作为冰冻圈微生物活动的热点, 为病毒提供了特殊的生存环境, 驱使病毒产生了生存与适应机制。通过综述国际上近些年来关于冰川表面冰尘洞内病毒的主要研究成果, 内容涉及冰尘洞在冰川生态系统中的重要性, 冰尘洞内病毒的丰度与生产力, 冰尘洞内病毒的遗传多样性, 病毒与宿主的相互作用关系, 以及病毒对于冰尘洞碳循环和食物网的重要性。此外, 还总结了当前冰尘洞内病毒研究存在的空白： 形态多样性、 降解率、 分离培养、 冰川表面散落冰尘内病毒, 指出了未来研究需要重点关注的几个方面, 为进一步研究病毒对于冰尘洞生态系统的重要影响提供科学参考, 并且对青藏高原冰川表面冰尘洞内病毒未来的研究提出了展望。     

广西养殖拟穴青蟹携带病原体状况的初步调查

采用微生物分离培养技术和巢式PCR技术相结合方法对2011年采自广西北海南万和古城、钦州茅岭和钦州港、防城港企沙和公车的拟穴青蟹(Scylla paramamosain)携带寄生虫、细菌和病毒状况开展调查.寄生虫检测结果显示,仅钦州茅岭拟穴青蟹鳃组织中发现寄生虫茗荷(儿).细菌性病原体检测结果显示,共有11种病原性细菌存在于健康养殖拟穴青蟹体内,弧菌属细菌性病原出现频率最高,其中副溶血弧菌和溶藻弧菌在调查区域内均有分布,其他菌种则呈现一定的地域特异性.病毒性病原体检测结果显示,白斑综合症病毒仅在钦州茅岭拟穴青蟹检出,阳性率为4.94%;双顺反子病毒于北海古城和钦州茅岭拟穴青蟹检出阳性携带率为3.70%;呼肠孤病毒携带率则较高,北海、防城港以及钦州拟穴青蟹均有检出,阳性率达25.93%.由此可见,10-11月广西区内健康养殖拟穴青蟹携带弧菌和呼肠孤病毒的比例远远高于其他病原体,为拟穴青蟹携带的主要病原体.     

流行性造血器官坏死病毒（EHNV）PCR快速检测试剂盒的研制

以流行性造血器官坏死病毒（EHNV）中的主衣壳蛋白基因保守区序列作为扩增靶序列,设计合成了一对特异性引物,应用快速的模板制备方法和优化PCR扩增条件,建立了EHNV病毒PCR快速检测技术,并开发成简便、快速、实用的检测试剂盒.该试剂盒的检测灵敏度相当于31个病毒粒子,模板制备时间约30 min,约4 h即可得到准确的结果;且无非特异性扩增带,适用于由EHNV病毒感染的鱼类苗种和水产品的检疫及水质环境的监测.     

Development and application of antibody microarray for white spot syndrome virus detection in shrimp

Detecting white spot syndrome virus (WSSV) in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based microarray is a novel proteomic technology that can meet the requirements. In this study, we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples. First, seven slides each with different modifications were characterized by atomic force microscope, and were compared in the efficiency of immobilizing proteins. Of the seven, 3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size. A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides, and then the microarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody. The results were measured by a laser chipscanner and analyzed with software. To obtain satisfied fluorescence signal intensity, optimal conditions were searched. The detection limit of the antibody microarray for WSSV is 0.62 μg/mL, with a proven long shelf life for 6 months at 4°C or 8 months at -20°C. Furthermore, concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection. These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV.     

Spatial interpolation and image-integrative geostatistical prediction of mosquito vectors for arboviral surveillance

Remote sensing can augment traditional methods of mosquito species surveillance for arboviruses. Abundance and patterns of mosquito vectors of West Nile virus in Chesapeake, Virginia, USA, were studied using light trap collection data and a Landsat-7 Enhanced Thematic Mapper+ digital image for spatial interpolation and geostatistical mapping of the abundance of 24 species of mosquitoes capable of transmitting West Nile virus to humans. We evaluated spatial interpolation techniques including inverse distance weighting, ordinary kriging, co-kriging geostatistics using combined Landsat-7 tasselled cap transform indices (brightness, greenness, and wetness) to characterize habitats and breeding conditions. Results highlight gaps in surveillance coverage, geostatistical improvement of vector patterns and abundance, and spatial patterns of error. Constraints and opportunities for adoption of remote sensing and spatial analysis for mosquito control are identified and discussed.     

H5、H7和H9亚型禽流感分子鉴别诊断方法的建立

为快速、敏感、特异地鉴别诊断现较为重要的禽流感病毒亚型,根据H5、H7和H9亚型禽流感病毒HA基因序列的保守区域设计出并合成3对特异性引物,利用这3对引物对H5、H7和H9亚型禽流感病毒的HA基因片断进行多联RT-PCR扩增,并对多联RT-PCR检测方法进行敏感性及特异性实验。结果表明:3对引物能特异性地扩增出H5、H7和H9亚型禽流感病毒HA基因片段,大小分别为490bp,365bp和686bp;该检测方法敏感性高,特异性强;初步建立起快速、敏感、特异地鉴别检测H5、H7和H9亚型禽流感病毒的分子诊断方法。     

Initiation of two ovarian cell lines from Fugu rubripes (Temminck et. Schlegel)

The ovary is an excellent system for studying stem cell renewal and differentiation, which is under the control of ovarian somatic cells. In order to understand oogenesis in Fugu rubripes (Temminck et. Schlegel) as a marine fish model of aquaculture importance, we established cell lines called TSOC1 and TSOC2 from a juvenile ovary of this organism. TSOC1 is composed of spindle epithelial-like cells, while the other is cobblestone-like cells. Therefore, TSOC1 and TSOC2 appear to consist of ovarian somatic cells. Growth requirement condition was investigated including temperature, concentration of FBS and pH. Significant fluorescent signals were observed after TSOC1 and TSOC2 cells were transfected with pEGFP-N3 vector, indicating its potential utility for genetic manipulation such as gene function studies. It is shown that these cell lines are effective for infection by the turbot reddish body iridovirus and flounder lymphosystis disease virus as evidenced by the appearance of cytopathic effect and virus propagation in the virus-infected cells, and most convincingly, the observation of viral particles by electron microscopy, demonstrating that TSOC1 and TSOC2 are suitable to study interactions between virus and host cells. It is believed that TSOC1 and TSOC2 will be useful tools to study sex-related events and interactions between primordial germ cells and oogonia cells during oogenesis. Therefore, establishment of ovary cell lines from Fugu rubripes seems to be significant for those research areas.     

“四深”微生物的地质作用——从气候环境变化到生态灾难

“四深”微生物是指深海、深地、深空和深时环境的微生物,特别是细菌、古菌、真菌、病毒等。人们对“四深”微生物的了解非常有限,是亟待突破的地球生物学前沿领域。“四深”微生物的研究对理解地球生命起源、界定生物圈的边界条件、促进地球科学与生命科学以及行星科学之间的交叉融合具有不可替代性的贡献。随着我国深海、深空、深地等重大工程计划的推进,一系列与“四深”微生物有关的前沿科学问题不断提出,包括地质微生物与气候环境的相互作用、地质微生物的生物安全与生态安全、地质微生物参与的隐匿地质过程等。特别是,“四深”环境活性氧自由基对微生物的影响、地质病毒对生物演化和地质过程的影响等前沿领域都亟待突破。活性氧自由基能对生物分子、细胞、组织和器官,乃至整个生物圈的演化以及微生物地质作用都产生重要影响。病毒引发了现代和近代诸多全球性疫情爆发,地质病毒则可能对生物的背景灭绝和大灭绝以及一些地质过程产生影响。     

微藻类病毒与宿主之间微生态关系

病毒感染、杀死和裂解微藻是海洋生态系统中的普遍现象,在自然海洋环境中,有多种因素可以导致浮游植物细胞的损失,其中微藻的自然死亡(即细胞裂解)率是导致微藻损失的一个重要因素.由病毒介导的宿主死亡不仅可影响藻类物种的种间演替,也可能会影响种内演替、藻类群落的丰度及多样性等.越来越多的证据表明,病毒可以通过减少宿主种群数量或防止藻类宿主种群数量达到高峰的方式来控制浮游植物动力学指标.因此,藻类病毒与宿主之间的相互作用在赤潮动力学和感染的传播中发挥了重要作用.同时,病毒具有高度特异性宿主范围的发现拓展了我们对微藻种群动力学过程的认识.本文从病毒-微藻稳定感染系统模型、病毒对微藻种群动力学的调节、病毒介导的微藻死亡、宿主对病毒侵染的防御以及影响病毒与宿主相互作用的环境因素等方面综述微藻类病毒与宿主的相互作用关系.     

Spatial autocorrelation of West Nile virus vector mosquito abundance in a seasonally wet suburban environment

The objective of this study is to quantify and model spatial dependence in mosquito vector populations and develop predictions for unsampled locations using geostatistics. Mosquito control program trap sites are often located too far apart to detect spatial dependence but the results show that integration of spatial data over time for Cx. pipiens-restuans and according to meteorological conditions for Ae. vexans enables spatial analysis of sparse sample data. This study shows that mosquito abundance is spatially correlated and that spatial dependence differs between Cx. pipiens-restuans and Ae. vexans mosquitoes.