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.      

“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。     

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。     

Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)

Viral hemorrhagic septicemia virus (VHSV) and marine birnavirus (MABV) are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification (LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms.     

Rhythmic swimming of the isopod Exosphaeroma obtusum (Dana)

The tidal swimming rhythm of the New Zealand rocky shore isopod Exosphaeroma obtusum (Dana) is described. Peak swimming was on the ebb tide about 2.5 h after high water; the peak in the dark period was 6–8 times greater in amplitude than the daytime peak. Positive thigmotaxis was shown at all other times. The rhythm period in continuous darkness (free‐running period) was greater than tidal periodicity, and there was a semi‐lunar swimming pattern with peaks at the first and third quarters of the moon. Rheotropic experiments showed that changes in water flow did not initiate the swimming rhythm. The work was compared with the infaunal species Pseudaega punctala and the northern Eurydice pulchra. It was concluded that infaunal species rely more on endogenous control of the swimming rhythm because of their isolation, when buried, from exogenous Zeitgeber.     

Modification of the polyethylene glycol 6000 precipitation method for recovering human and indicator viruses from oysters and mussels

Human viruses are a common contaminant of shellfish affected by human sewage wastes. They are difficult to detect as they are not easily separated from shellfish tissue. This paper describes a modification of the polyethylene glycol (PEG) precipitation technique for recovery of enteroviruses and F‐specific bacteriophages from the Pacific oyster (Crassostrea gigas) and the green‐lipped mussel (Perna canaliculus). Modifications adopted were the use of only the digestive gland tissue for virus extraction, resuspension of the primary PEG pellet in 4 volumes of eluent, and the introduction of a secondary PEG precipitation to reconcentrate the virus containing extract. The recovery rate of the virus extraction process was not affected by introduction of the secondary concentration step (overall recovery remained at 60–70% of the virus input). The advantages of reduction of tissue residue in the extract, smaller final volume, and the ability to process 2–3 times the number of individual shellfish for the same effort, improve the practicality of the method.