水环境中致病菌分子生物学检测技术研究进展

总结了水环境中主要致病菌及其导致的疾病；介绍了检测水环境样品中致病菌的聚合酶链式反应（PCR）、实时定量PCR、等温扩增技术、生物传感器和高通量测序等分子生物学技术研究进展；分析了不同检测技术的优缺点和应用特点；提出了水环境中致病菌分子生物学检测技术的未来发展方向，为水环境中致病菌分析检测和风险控制提供研究思路和技术支撑。     

基于改进的多目标决策的水环境质量综合评价

在对水环境质量综合评价中,多目标决策-理想区间法解决了水环境质量评价标准是区间而非点的缺陷。但是在计算监测点到各理想区间向量的距离时,各水环境质量指标权重直接影响综合评价的结果,通常的确定方法是简单的假设各水环境质量指标的权重相等,这与实际情况相悖。为了解决这一问题,提出了将超标法用于多目标决策法中,利用超标法确定各水环境质量指标的权重,然后将其应用于多目标决策-理想区间法来分析水环境质量等级。并将改进后的多目标决策-理想区间法应用于珠江口及邻近海域的水环境中。基于超标法确定权重的多目标决策-理想区间法与聚类分析相比更有效,与等权重的多目标决策-理想区间法相比,更能体现水环境的污染状况,可应用于各种环境因子的综合评价中。     

水环境生物监测的发展方向与核心技术

水环境生物监测是环境监测的重要内容,它应重点说清环境胁迫的生物效应。简述了总量管理、流域管理、风险管理、生态管理等环境管理对水环境生物监测有迫切需求,应引入＂生态系统健康＂、＂生物完整性＂、＂环境胁迫＂、＂全排水毒性＂等现代环境生物监测的基本概念,建立水环境生物监测技术发展的理论基础,发展生物完整性、综合毒性等监测与评价核心技术;革新现行监测方法体系,建立包括QA/QC、快速方法等支持系统在内的现代水环境生物监测业务化方法体系;创新评价技术体系,建立水环境生态健康评价及综合毒性评价指标体系、基准及分级管理标准,确立水环境质量管理的生物学目标。     

滇池流域水环境综合管理技术支撑平台构建研究

滇池流域水环境综合管理技术支撑平台以实现水环境数据信息化、支撑多元化、管理智能化为目标,集成水环境多元数据采集传输、融合共享及动态表征技术,突破高原湖泊流域多目标复杂环境综合管理决策控制技术,构建"数据中心-业务系统-信息发布"为主线的综合管理平台,其以水环境数据中心为支撑,以水环境信息系统与水环境专家决策支持系统为核心,以水环境信息发布系统为共享交换媒介,为流域管理提供监控预警、总量控制、预测预报、项目评估及应急决策等技术服务,有效支撑滇池流域水环境监控预警和综合管理。     

以嘉陵江铊浓度异常事件为例探讨《重特大突发水环境事件应急监测工作规程》在应急监测中的应用

以"1.20"嘉陵江铊浓度异常事件应急监测为例,阐述了《重特大突发水环境事件应急监测工作规程》在应急监测工作中的应用。通过实例分析了应急监测全过程中的监测方案编制、样品采集安排、监测实验室设置、监测方法选择、监测结果报送、污染趋势预测以及应急状态终止等内容,结果表明《重特大突发水环境事件应急监测工作规程》能够指导应急监测工作顺利开展,该案例可为相关突发水环境事件的现场处置和应急监测工作提供技术参考。     

PCR技术对水中病原体的检测

水中有病毒、细菌、原生动物、蠕虫等多种病原体。常规的检测方法是用大肠菌作为病原体的指示生物,但是大肠菌的生存特性与病毒、原生动物差异较大,难以指示病毒、原生动物的存在与否。对每一种病原体单独进行检测,费时、费力。由于PCR技术具有快速、灵敏、特异的特点,使得它对于水中病原体的检测具有较高的应用价值。     

国家水环境质量监测网络发展历程与展望

阐述了水环境质量监测的定义和目的，介绍了构成国家水环境质量监测网络的6个子网，即地表水环境质量监测网、地表水环境质量自动监测网、地下水环境质量监测网、饮用水源地环境质量监测网、水污染防治专项规划水体环境质量监测网和锰三角地区水环境质量监测网的发展历程与现状。指出了目前该网络存在的不足，提出建立水环境质量监测网络设计技术体系、扩大监测网覆盖面、扩展监测项目等建议。     

饮用水源微生物快速检测技术的发展及应用

微生物是威胁饮用水安全的首要问题，水环境微生物快速检测技术的开发和应用是推动饮用水源微生物快速检测和水质安全预警技术发展的保障。随着对水质微生物污染快速检测和准确预警新要求的提出，水环境中微生物在线检测和预警技术得到了越来越多的开发和应用。笔者总结了水环境常见微生物检测方法和技术的发展，重点讨论了饮用水源微生物快速检测技术的发展和应用，根据各项技术的应用和推广使用程度，将其归纳为常用快速检测技术、潜在适用快速检测技术和新型快速检测技术等类别，并详细阐述了一些应用较广的技术，以期为构建水质微生物污染早期预警系统提供参考。     

水环境中PAHs源解析研究方法比较

综述了近年来水环境中PAHs源解析的方法,包括化学质量平衡法、因子分析法、稳定碳同位素法和比值法等,重点评述了方法的理论基础、适用条件、应用状况和存在问题,并结合我国情况分析了水环境中PAHs源解析方法的发展趋势.认为因子分析法与多元统计方法结合使用是当前我国PAHs源解析的最适用方法,可以实现对源贡献率的定量分析,与GIS技术结合应用,能实现PAHs源的空间统计分析,反映污染源影响作用的空间分布规律.     

流式细胞术在水质检测领域的研究进展

流式细胞术作为一种新兴技术,具有检测快速简便、灵敏度高等优点,在水环境突发污染事件及水处理工艺的评价中正发挥着重要作用。随着荧光染色及配套技术的突破性发展,流式细胞术在水环境领域的研究和应用得到逐步拓展。笔者系统评述了流式细胞术检测原理及技术特点、水环境检测影响因素及其应用进展等,并对该技术在水质检测评价中的推广应用提出建议。     

Ultracentrifugation as a direct method to concentrate viruses in environmental waters: virus-like particle enumeration as a new approach to determine the efficiency of recovery

Some health important enteric viruses are considered to be emerging waterborne pathogens and so the improvement of detection of these viruses in the aquatic environment is one of the most important steps in dealing with these pathogens. Since these viruses may be present in low numbers in water, it is necessary to concentrate water samples before viral detection. Although there are several methods to concentrate viruses in environmental waters, all present some drawbacks and consequently the method should be chosen that, despite its limitations, is adequate to achieve the aim of each study. As the effectiveness of the concentration methods is evaluated by determining the efficiency of viral recovery after concentration, it is important to use a simple and effective approach to evaluate their recovery efficiency. In this work ultracentrifugation, usually used as a secondary step for virus concentration, was evaluated as the main method to concentrate directly viruses in environmental water samples, using the microscopic enumeration of virus-like particles (VLP) as a new approach to estimate the efficiency of recovery. As the flocculation method is currently employed to concentrate viruses in environmental waters, it was also used in this study to assess the efficiency of the ultracentrifugation as the main viral concentration method in environmental waters. The results of this study indicate that ultracentrifugation is an adequate approach to concentrate viruses directly from environmental waters (recovery percentages between 66 and 72% in wastewaters and between 66 and 76% in recreational waters) and that the determination of VLP by epifluorescence microscopy is a simple, fast and cheap alternative approach to determine the recovery efficiency of the viral concentration methods.     

Molecular detection of three gastroenteritis viruses in urban surface waters in Beijing and correlation with levels of fecal indicator bacteria

To assess the presence of three gastroenteritis viruses responsible for human acute gastroenteritis in surface water, a 1-year study was carried out in the city of Beijing, China. A total of 108 urban surface water samples were collected from nine collection sites which were defined with a global positioning system in rivers or lakes from September 2006 to August 2007. The water samples were subjected to virus concentration using an HA electronegative filter, followed by polymerase chain reaction (PCR) for rotavirus (RV) astrovirus (AV), and norovirus (NV). It showed that the number of viruses detected in water samples from different sites was variable, totaling 63 virus strains, with rotavirus (48.1%) verified as the most prevalent detected, followed by astrovirus (AV, 5.6%), and norovirus (NV, 4.6%). RV was also quantified by real-time PCR and the concentration of RV ranged from 0 to 18.27 genome copies·L(-1). And the distributions of RV in surface water were abundant in cold weather (from September to February) while less prevailing in warm weather (from March to August). The high detection rate of RV we encountered in this study provided convincing evidence that RV circulated at a certain frequency in the Beijing population. There was no statistically significant correlation between RV levels and both fecal coliform (R (2)?=?0.02) and Enterococcus faecalis (R (2)?=?0.02) densities. Our study suggests prolonged virus persistence in aquatic environments and emphasizes the enteric virus group as the most reliable for environmental monitoring.     

Evaluation of concentration efficiency of the Pseudomonas aeruginosa phage PP7 in various water matrixes by different methods

Enteric viruses monitoring in surface waters requires the concentration of viruses before detection assays. The aim of this study was to evaluate different methods in terms of recovery efficiencies of bacteriophage PP7 of Pseudomonas aeruginosa, measured by real-time PCR, using it as a viral control process in water analysis. Different nucleic acid extraction methods (silica–guanidinium thiocyanate, a commercial kit (Qiagen Viral RNA Kit) and phenol–chloroform with alcohol precipitation) exhibited very low recovery efficiencies (0.08–4.18 %), being the most efficient the commercial kit used for subsequent experiments. To evaluate the efficiency of three concentration methods, PBS (as model for clean water) and water samples from rivers were seeded to reach high (HC, 10⁶ pfu ml^?1) and low concentrations (LC, 10⁴ pfu ml^?1) of PP7. Tangential ultrafiltration proved to be more efficient (50.36?±?12.91, 17.21?±?9.22 and 12.58?±?2.35 % for HC in PBS and two river samples, respectively) than adsorption–elution with negatively charged membranes (1.00?±?1.34, 2.79?±?2.62 and 0.05?±?0.08 % for HC in PBS and two river samples, respectively) and polyethylene glycol precipitation (15.95?±?7.43, 4.01?±?1.12 and 3.91?±?0.54 %, for HC in PBS and two river samples, respectively), being 3.2–50.4 times more efficient than the others for PBS and 2.7–252 times for river samples. Efficiencies also depended on the initial virus concentration and aqueous matrixes composition. In consequence, the incorporation of an internal standard like PP7 along the process is useful as a control of the water concentration procedure, the nucleic acid extraction, the presence of inhibitors and the variability of the recovery among replicas, and for the calculation of the sample limit of detection. Thus, the use of a process control, as presented here, is crucial for the accurate quantification of viral contamination.     

Molecular approaches to microbiological monitoring: fecal source detection

Molecular methods are useful both to monitor natural communities of bacteria, and to track specific bacterial markers in complex environments. Length-heterogeneity polymerase chain reaction (LH-PCR) and terminal restriction fragment length polymorphism (T-RFLP) of 16S rDNAs discriminate among 16S rRNA genes based on length polymorphisms of their PCR products. With these methods, we developed an alternative indicator that distinguishes the source of fecal pollution in water. We amplify 16S rRNA gene fragments from the fecal anaerobic genus Bacteroides with specific primers. Because Bacteroides normally resides in gut habitats, its presence in water indicates fecal pollution. Molecular detection circumvents the complexities of growing anaerobic bacteria. We identified Bacteroides LH-PCR and T-RFLP ribosomal DNA markers unique to either ruminant or human feces. The same unique fecal markers were recovered from polluted natural waters. We cloned and sequenced the unique markers; marker sequences were used to design specific PCR primers that reliably distinguish human from ruminant sources of fecal contamination. Primers for more species are under development. This approach is more sensitive than fecal coliform assays, is comparable in complexity to standard food safety and public health diagnostic tests, and lends itself to automation and high-throughput. Thus molecular genetic markers for fecal anaerobic bacteria hold promise for monitoring bacterial pollution and water quality.     

水中烷基汞检测技术的研究进展

简述了水中烷基汞的分析测定技术,包括富集材料的种类和性能,水样富集处理方法,色谱和毛细管电泳分离技术,光谱、质谱及其他多种检测技术的联合应用等,并对水中烷基汞的分析检测技术发展进行了展望。     

水中亚硝胺类消毒副产物的污染现状、形成与控制

综述了水中亚硝胺类消毒副产物的来源、浓度水平及限定标准,以及亚硝胺类物质(NAs)的生成机制、分析检测技术、控制与去除方法。指出,随着人们对饮用水安全消毒的重视和污水消毒的推广,应增加大范围水中NAs污染水平的调查工作,以及对复杂污水体系中NAs生成前体物的研究,优化和改进消毒工艺,减少NAs的生成,针对复杂水样建立准确可靠的NAs检测方法。     

Development of an extraction method for perchlorate in soils

Perchlorate originates as a contaminant in the environment from its use in solid rocket fuels and munitions. The current US EPA methods for perchlorate determination via ion chromatography using conductivity detection do not include recommendations for the extraction of perchlorate from soil. This study evaluated and identified appropriate conditions for the extraction of perchlorate from clay loam, loamy sand, and sandy soils. Based on the results of this evaluation, soils should be extracted in a dry, ground (mortar and pestle) state with Milli-Q water in a 1 ratio 1 soil ratio water ratio and diluted no more than 5-fold before analysis. When sandy soils were extracted in this manner, the calculated method detection limit was 3.5 microg kg(-1). The findings of this study have aided in the establishment of a standardized extraction method for perchlorate in soil.