噬菌体MS2生物学特性研究

本文分析了噬菌体MS2在水质监测中作为水中肠道病毒指示生物的可行性与优势,并通过试验测定了噬菌体MS2的热稳定性、pH值稳定性、最佳感染复数和一步生长曲线等生物学参数,结果表明:噬菌体MS2的热稳定性较弱,在70℃时5 min内即会全部失活;在pH值4～10的范围内噬菌体MS2均可稳定存在;噬菌体MS2感染其宿主大肠杆...     

The Effect of Heat on the Physicochemical Properties of Bacteriophage MS2

The differences in physicochemical characteristics between infectious and non-infectious viral particles are poorly known. Even for heat, which is known as one of the most efficient treatments to inactivate enteric viruses, the global inactivation mechanisms have not been described yet. Such knowledge would help distinguish between both types of particles and therefore clarify the interpretation of the presence of viral genomes in food after heat treatment. In this study, we examined in particular the differences in electrostatic charge and hydrophobicity between the two particle types. MS2 phage, a common surrogate for enteric viruses, was used as a model virus. The heat-induced inactivation process of the infectious phages caused hydrophobic domains to be transiently exposed and their charge to become less negative. The particles also became progressively permeable to small molecules such as SYPRO Orange dye. The presence of non-infectious phage particles in which the genome was not accessible to RNases has been clearly demonstrated. These observations were done for MS2 phages exposed to a temperature of 60 °C. When exposed to a temperature higher than their critical temperature (72 °C), the particles were disrupted and the genome became available for RNases. At lower temperatures, 60 °C in this study, the transient expression of hydrophobic domains of remaining infectious phages appeared as an interesting parameter for improving their specific detection.     

藻细胞和高岭土的存在对病毒MS2存活的影响

选用病毒MS2作为水中肠道病毒的指示病毒,高岭土和铜绿微囊藻分别作为无机颗粒物和有机颗粒物,研究颗粒物浓度、pH值、不同价态离子浓度、天然有机物(NOM)等水质条件下,无机(高岭土)、有机(铜绿微囊藻)颗粒物存在对病毒MS2存活的影响.结果表明,无机颗粒物高岭土对病毒MS2的存活无明显影响,但当水体钙硬度(钙离子产生的硬度)较大时,病毒MS2的表观存活量增加1个对数;铜绿微囊藻的存在会导致病毒MS2的存活量降低1个对数左右,但当溶液的pH值大于4.0或铜绿微囊藻的浓度小于1.0×106cells·L-1时,藻类对病毒的生存无明显影响;当水体钙硬度较大时,藻反而会增加病毒MS2的存活对数.因此在高浊水、高藻水中,水的钙硬度增加会使水体中病毒生存能力变强,进而增加饮用水的安全风险.     

Impact of Sodium Chloride, Sucrose and Milk on Heat Stability of the Murine Norovirus and the MS2 Phage

Until now, little is known about the influence of food additives on heat inactivation of noroviruses. Only a few studies have shown a protective or inhibiting effect on virus infectivity caused by the food matrix. Therefore, the aim of this study was to examine the influence of sodium chloride, sucrose and milk on heat stability of the surrogates murine norovirus (MNV) and MS2 phage at 60 °C for 1–5 min in PBS for MNV and for 5–120 min in suspension medium buffer for MS2 phage. Different concentrations of sodium chloride (5, 10 %) and sucrose (5, 50 %) were added to the respective buffers. In addition, commercially available milk with different fat concentrations (0.3, 1.5, 3.5 %) was investigated in this study. In general, a linear titre reduction for MNV and MS2 phage could be observed, except for the heat treatment of MNV in PBS with 50 % sucrose. A protective effect of PBS with 50 % sucrose and of the matrix milk on MNV could be concluded. All other tested conditions did not show any influence on virus inactivation. However, MS2 phage did show a higher heat resistance throughout the experiments compared to MNV. In future investigations, it should be tested, whether the achieved data may be considered in risk assessments of heat-treated food products with high concentrations of sugar. Furthermore, it should be clarified, whether these results can also be referred to complex food matrices.     

理化因素及感染复数对噬菌体MS2活性的影响

噬菌体MS2作为潜在的水中肠道病毒指示生物,已逐渐成为回用水微生物学安全评价指标的主要研究方向之一。测定了温度、pH值、紫外线和感染复数对噬菌体MS2活性的影响。结果表明:噬菌体MS2在70℃时5 min内即会全部失活,在pH值4～10的范围内均可稳定存在,在短波紫外线照射下25 min内即可失活;噬菌体MS2感染其宿主大肠杆菌的最佳感染复数为1。     

Evaluation of Viral Recovery Methodologies from Solid Waste Landfill Leachate

Food and Environmental Virology - Leachate from solid waste landfill is a dark liquid of variable composition and possible source of contamination of groundwater and surface waters. This study aims...     

Potential of Pulsed Light to Inactivate Bacteriophage MS2 in Simple Liquid Medium and on Complex Foodstuffs

The virucidal efficacy of a pulsed light treatment applied to viral suspensions, glass beads and herb powders was studied for the F-RNA bacteriophage MS2. The experimental results obtained demonstrated the high potential of this technology to efficiently decontaminate simple matrices but underlined the complexity of application to complex food matrices.     

紫外活化过硫酸钠灭活水中噬菌体MS2的特性及机制

水环境中的病毒对常见的消毒技术有较强的抵抗力.为了开发水中病毒的高效灭活技术,以噬菌体MS2为对象,研究紫外活化过硫酸钠（UV/PS）体系灭活病毒的特性和机制.采用双层平板法对噬菌体MS2进行定量检测,研究UV/PS对水样中噬菌体MS2的灭活率和动力学特征,并考察PS用量、pH值和噬菌体初始浓度等因素对灭活效果的影响.利用透射扫描电镜观察UV/PS处理前后噬菌体的形貌,利用电子顺磁共振波谱法确认反应体系中存在的自由基种类.在自由基淬灭实验的基础上,分析计算UV/PS体系中各因素对噬菌体灭活的贡献率.结果表明,当紫外辐照强度为160 μW·cm^-2时,UV/PS处理4 min即可去除4.39 lg的噬菌体MS2,较单独使用同样辐照剂量的UV消毒灭活率高1.44 lg.UV/PS体系对噬菌体MS2的灭活动力学过程符合一级反应动力学模型.增加体系中的PS初始浓度能明显提高对噬菌体的灭活率和灭活速率,而pH和噬菌体初始浓度对UV/PS灭活噬菌体的影响较小.UV/PS处理可导致噬菌体的衣壳破损,促进了噬菌体颗粒团聚.UV/PS体系中存在SO₄^-·和·OH,是噬菌体MS2灭活的重要因素.·OH比SO₄^-·对噬菌体MS2灭活的贡献更大.     

同时提取土壤微生物DNA和RNA方法的研究

获得高浓度、大片段、多样性程度高的土壤微生物总DNA是研究土壤微生物群落结构的分子生态学基础.研究采用PBS缓冲液洗涤土壤样品,结合SDS裂解微生物细胞的方法,同时提取分别添加小麦秸秆粉和油菜秸秆粉的两种土壤样晶的微生物DNA和RNA.对提取出的DNA和RNA进行酶切,基因组PCR和反转录PCR,实验结果表明该法提取的...     

Assessment of Viral Contamination of Five Brazilian Artisanal Cheese Produced from Raw Milk: a Randomized Survey

Food and Environmental Virology - Enteric viruses have been described as important contaminants in fresh and ready-to-eat foods such as sandwiches, deli meat and dairy products. This is a...     

Evaluating Efficacy of Field-Generated Electrochemical Oxidants on Disinfection of Fomites Using Bacteriophage MS2 and Mouse Norovirus MNV-1 as Pathogenic Virus Surrogates

Surface disinfection, as part of environmental hygiene practices, is an efficient barrier to gastroenteritis transmission. However, surface disinfectants may be difficult to obtain in remote, resource-limited, or disaster relief settings. Electrochemical oxidants (ECO) are chlorine-based disinfectants that can be generated using battery power to electrolyze brine (NaCl) solutions. Electrolysis generates a mixed-oxidant solution that contains both chlorine (HOCl, OCl^?) and reactive oxygen species (e.g., ·OH, O₃, H₂O₂, and ·O_2?) capable of inactivating pathogens. One onsite generator of ECO is the Smart Electrochlorinator 200 (SE-200, Cascade Designs, Inc.). In a laboratory study, we assessed ECO surface disinfection efficacy for two gastrointestinal virus surrogates: bacteriophage MS2 and murine norovirus MNV-1. We quantified both infectivity and nucleic acid inactivation using culture-dependent and independent assays. At free available chlorine concentrations of 2,500 ppm and a contact time of 30 s, ECO inactivation of infective MS2 bacteriophage exceeded 7 log₁₀ compared to MNV-1 disinfection of approximately 2 log₁₀. Genomic RNA inactivation was less than infective virus inactivation: MS2 RNA inactivation was approximately 5 log₁₀ compared to MNV-1 RNA inactivation of approximately 1.5 log₁₀. The results are similar to inactivation efficacy of household bleach when used at similar free available chlorine concentrations. Our work demonstrates the potential of ECO solutions, generated onsite, to be used for surface disinfection.     

Use of F-Specific RNA Bacteriophage to Estimate Infectious Norovirus Levels in Oysters

Food and Environmental Virology - Contamination of bivalve shellfish, particularly oysters, with norovirus is recognised as a significant food safety risk. Methods for quantification of norovirus...     

Persistence of MS-2 Bacteriophage Within Eastern Oysters

Male-specific bacteriophages have been proposed as human enteric virus indicators for shellfish. In this study, Eastern oysters (Crassostrea virginica) were individually exposed to 5.6 × 10¹⁰ PFU of MS-2 for 48 h at 15 °C followed by collective maintenance in continuously UV-sterilized seawater for 0–6 weeks at either 7, 15, or 24 °C. Initial contamination levels of MS-2 were >6 log PFU. Assessment of weekly declines of viable MS-2 indicated that cooler temperatures dramatically enhanced the persistence of MS-2 within oyster tissues. At 3 weeks, the average log PFU reductions for MS-2 within oysters were 2.28, 2.90, and 4.57 for oysters held at 7, 15, and 24 °C, respectively. Fitting temporal survival data with linear and nonlinear Weibull models indicated that the Weibull model best fit the observed reductions. In total, these data can serve as a guideline for regulatory agencies regarding the influence of water temperature on indicator phage after episodic sewage exposure.     

活性污泥对病毒的生物吸附特性

针对污水处理过程中可能存在的病原微生物对人体健康的风险,以f2噬菌体为模型病毒,采用静态吸附方法,研究病毒在活性污泥上的生物吸附特性及其影响因素,以期了解病毒在活性污泥处理系统中固液两相体系的分配规律.结果表明,活性污泥对病毒的吸附速率快,吸附量大,在120 min时即能达到吸附平衡,平衡吸附率高达96%以上,对病毒的去除效果较为理想;该吸附符合Freundlich吸附等温式,且吸附率随着病毒浓度的增加而呈现出下降趋势;另外,pH值、金属阳离子浓度及有机物浓度均可显著影响病毒在活性污泥上的吸附.     

The Impact of the Extreme Amazonian Flood Season on the Incidence of Viral Gastroenteritis Cases

During the Amazonian flood season in 2012, the Negro River reached its highest level in 110 years, submerging residential and commercial areas which appeared associated with an elevation in the observed gastroenteritis cases in the city of Manaus. The aim of this study was to evaluate the microbiological water quality of the Negro River basin during this extreme flood to investigate this apparent association between the illness cases and the population exposed to the contaminated waters. Forty water samples were collected and analysed for classic and emerging enteric viruses. Human adenoviruses, group A rotaviruses and genogroup II noroviruses were detected in 100, 77.5 and 27.5% of the samples, respectively, in concentrations of 10³–10⁶ GC/L. All samples were compliant with local bacteriological standards. HAdV2 and 41 and RVA G2, P[6], and P[8] were characterised. Astroviruses, sapoviruses, genogroup IV noroviruses, klasseviruses, bocaviruses and aichiviruses were not detected. Statistical analyses showed correlations between river stage level and reported gastroenteritis cases and, also, significant differences between virus concentrations during this extreme event when compared with normal dry seasons and previous flood seasons of the Negro River. These findings suggest an association between the extreme flood experienced and gastrointestinal cases in the affected areas providing circumstantial evidence of causality between the elevations in enteric viruses in surface waters and reported illness.     

环境因子对舰载机金属构件维修检查间隔期的影响

结合金属构件腐蚀的影响因素和原理,定义了海洋环境因子;研究了海洋环境对舰载机金属构件维修检查间隔期的影响,并通过例证对舰载机金属构件的维修检查间隔期进行了调整,为舰载机在海洋环境下金属构件的维修决策提供了可靠的依据.     

化学沉淀法回收污泥中氮磷的影响因素研究

污水处理厂污泥上清液中富集着较高浓度溶解性的氮磷,将此部分氮磷形成磷酸盐沉淀(如磷酸氨镁、磷酸钙、磷酸铝等)加以回收利用,受到各种因素的影响.文章以正交试验得出的影响因素为基础,深入研究了pH、初始PO43--p的浓度、Mg/P和反应时间对某污水厂污泥上清液中磷酸氨镁沉淀法回收氮磷的影响.结果表明:pH是影响污泥上清液...     

An Optimised Direct Lysis Method for Viral RNA Extraction and Detection of Foodborne Viruses on Fruits and Vegetables

Detection of norovirus (NoV) and hepatitis A virus (HAV) on fruits and vegetables using current standard methodologies can be inefficient. Method optimisat     

电镀污泥中镍的回收技术研究

采用30％N902从除杂后的电镀污泥氨浸液中回收金属镍。在萃取原料液pH=9,相比（A／O）=2：1,反应时间为5min条件下可使镍的萃取率达到99％。负载有机相经水洗后,用2mol／L A／O=1：1的硫酸进行反萃,反萃时间为30min,反萃级数为8级,得到产品硫酸镍。硫酸镍溶液中镍离子含量〉90g／L,其它杂质达到产品质量要求。