粪大肠菌群多管发酵测定法的改进研究

对地表水和废水中粪大肠菌群目前通用的多管发酵检测方法进行优化。将水样接种至乳糖蛋白胨培养液后,在44.5℃±0.5℃培养箱中培养24h记录结果,并与优化前的方法进行对比。结果表明,改进后的方法在时间上比改进前节省了约24~48小时,但两者在结果上无统计学意义的差别。多管发酵改进法可以用于地表水和废水中粪大肠菌群的检测。     

荧光法快速测定水中粪大肠菌群

为研究荧光法测定水中粪大肠菌群的适用性,在6家实验室使用荧光法对粪大肠菌群的标准菌株和实际水样进行测定,并与多管发酵法进行比对。结果表明:6家实验室对标准菌株的监测结果均在误差范围内,实验室内相对误差为-2. 75%～1. 71%,准确度较高;对3个不同浓度实际水样进行精密度测定,实验室内相对标准偏差分别为1. 30%～3. 93%、1. 94%～4. 72%、1. 88%～4. 54%,实验室间的相对标准偏差分别为1. 14%、1. 59%、1. 72%,精密度较高;用荧光法与多管发酵法同时测定实际水样,2种方法测试结果线性相关性良好且变化趋势相同,对测试数据进行t检验,2种方法测试结果无显著差异。     

A Regional Survey of the Microbiological Water Quality along the Shoreline of the Southern California Bight

A regional survey of the microbiological water quality along the shoreline of the Southern California Bight (SCB), from Point Conception south to Ensenada, Mexico, was conducted during August, 1998, by 36 agencies under the coordination of the Southern California Coastal Water Research Project (SCCWRP). Microbiological water quality was assessed by calculating the percentage of shoreline-mile-days that exceeded bacterial indicator thresholds for total and fecal coliforms, total/fecal ratios, and enterococci. Sample sites were selected using a stratified random sampling approach, with the SCB recreational shoreline divided into six strata: high- and low-use sandy beaches, high- and low-use rocky shoreline, and perennial and ephemeral freshwater outlets. Samples were collected on a weekly basis at a total of 253 sites, beginning on August 2^nd, 1998 and continuing for five weeks. Samples were analyzed by 22 participating labs using their normal methods (multiple tube fermentation, membrane filtration, Colilert® and/or Enterolert®). All labs met testing criteria established through intercalibration exercises and quality control check samples distributed during the sampling period. Nearly 95% of the shoreline-mile days did not exceed daily and monthly bacterial indicator thresholds, demonstrating good bacteriological water quality along the SCB shoreline. Freshwater outlets, comprised mainly of storm drains, had the poorest water quality with 60% and 40% of the shoreline-miles exceeding monthly and daily thresholds, respectively. Freshwater outlets were also more likely to demonstrate exceedances by multiple indicators at a single site, and repeat exceedances at sites over the five-week period. Compared with the southern California beaches, Mexican beaches had nearly 5 times the number of exceedances for total and fecal coliforms, and nearly 8 times the number of exceedances for total/fecal ratios.     

A Comparison of Bacterial Indicators and Methods in Rural Surface Waters

The United States Environmental Protection Agency (USEPA) recommends the use of Escherichia coli (E. coli) and enterococci as indicators of enteric pathogens in fresh waters; however, fecal coliform analyses will remain important by virtue of the large amount of historic data collected in prior years. In this study, we attempted, in a real-world situation (i.e., a rural inland watershed in the Piedmont of South Carolina) to compare different bacterial indicators and methods to one another. We compared fecal coliforms, enumerated by membrane filtration with E. coli, enumerated by a commercialized enzyme substrate method and observed E. coli/fecal coliform ratios of 1.63 and 1.2 for two separate tests. In the same watershed, we observed an E. coli/fecal coliform ratio of 0.84 when we used the commercialized enzyme substrate method for both enumerations. Given these results, users of such data should exercise care when they make comparisons between historic membrane filtration data and data acquired through the use of the more modern enzymatic methods. Some sampling and side-by-side testing between methods in a specific watershed may be prudent before any conversion factors between old and new datasets are applied.     

水中粪大肠菌群检测方法的比较

采用标准菌株、实际水样和国际标准样品，比较纸片快速法与多管发酵法的一致性。标准菌株试验表明，两种方法在粪大肠菌群的定性检测上没有显著性差异；实际水样试验表明，纸片快速法的检测结果略低于多管发酵法，但两种方法检测结果的回归关系显著；国际标准样品试验表明，两种方法的精密度与准确度均无统计学意义上的显著性差异。     

酶底物法检测地表水粪便污染的适用性探讨

为探讨酶底物法检测地表水粪便污染的适用性，采用酶底物法与多管发酵法同步检测了长江南京段、秦淮河、玄武湖等水中大肠菌群。结果显示：在多数情况下，酶底物法检测结果略低于多管发酵法，两种方法没有统计学差异（P〉0．05）；与多管发酵法相比，酶底物法检测粪便污染特异性强、时间周期短、二次污染小、简便快速，适于推广应用。     

Environmental pollutions impacts on the bacteriological and physicochemical quality of suburban and rural groundwater supplies in Marrakesh area (Morocco)

This study scrutinized bacteriological and chemical quality of groundwater supplies of Marrakesh (Morocco) within a year. It assessed the influence of some chemical factors on fecal and opportunistic pathogenic bacterial communities. The annual average densities of fecal coliforms, fecal streptococci and Pseudomonas aeruginosa were respectively: 1891 colony forming units (CFU)/100 mL, 1246 CFU/100 mL and 206 CFU/100 mL. The total occurrence of these bacteria during the period of study was 94%. Detectable non-O1 Vibrio cholerae was present in 81% of samples and the mean abundances ranged from 0 to 11100 MPN/100 mL. Significant correlations between fecal coliforms and streptococci and between fecal coliforms and non-O1 V. cholerae (p < 0.01) were found. Fecal coliforms can be used to detect the presence of non-O1 V. cholerae in this groundwater. These well waters were greatly mineralized, hard, salt with high concentrations of nitrogenous ions and major elements (Ca(2+), Na(+), Cl(-), SO(2-)(4) and K(+)). Most of chemical parameters were relatively steady with time except for calcium, ammonium, nitrites and organic matter which were seasonally influenced. The degree of correlation between chemical parameters and the abundances of isolated bacteria is heterogeneous. The concentrations of calcium and nitrites favored (p < 0.01) the abundance of fecal coliforms and streptococci, but reduced P. aeruginosa abundances. Calcium, salinity, chlorides, nitrites and nitrates tests can be used for screening in situ when the laboratory facilities are limited. This work is expected to assist local authorities in developing plans and actions to reduce the pollution to acceptable levels.     

粪大肠菌群酶底物法在环境监测中的应用研究

粪大肠菌群作为水体粪便污染的指示菌，对地表水的水质监测评价具有重要作用。采取较为精确、快速的粪大肠菌群检测方法，对控制流行疾病的发生和传播有重要的科学意义。从粪大肠菌群的检测意义、监测方法、标准化应用情况等方面进行归纳分析，提出推广和标准化酶底物法监测粪大肠菌群的迫切性。     

Making performance-based chemistry work: how we created comparable data among laboratories as part of a Southern California marine regional assessment

Quality assurance procedures to ensure consistency among chemistry laboratories typically involves the use of standard methods and state certification programs that require laboratories to demonstrate their ability to attain generic performance criteria. To assess whether these procedures are effective for ensuring comparability when processing local samples with potentially complex matrices, seven experienced, state-certified laboratories participated in an intercalibration exercise. Each laboratory was permitted to use their typical methodology for quantifying PAH, PCB and DDT on shared samples collected from Santa Monica Bay and the Palos Verdes Shelf, two sites with a complex mix of constituents. In the initial intercalibration exercise, results from these laboratories differed by as much as an order of magnitude for all three chemical groups. Much, but not all, of the difference was attributable to differences in detection capability. A series of studies was conducted to identify the reasons for the observed differences, which varied among laboratories and included methodological differences, instrument sensitivity differences, and differing interpretations of chromatograms. Following these investigations and resulting modifications to laboratory procedures, the exercise was repeated. The average coefficient of variation among laboratories across all chemical parameters was reduced to less than 30%. Our results suggest that performance-based chemistry can produce comparable results, but the certification processes presently in place that focus on general laboratory procedures and simple matrices are insufficient to achieve comparability.     

Feasibility of the hydrogen sulfide test for the assessment of drinking water quality in post-earthquake Haiti

In 2010, a magnitude 7.0 earthquake struck Haiti, severely damaging the drinking and wastewater infrastructure and leaving millions homeless. Compounding this problem, the introduction of Vibrio cholerae resulted in a massive cholera outbreak that infected over 700,000 people and threatened the safety of Haiti’s drinking water. To mitigate this public health crisis, non-government organizations installed thousands of wells to provide communities with safe drinking water. However, despite increased access, Haiti currently lacks the monitoring capacity to assure the microbial safety of any of its water resources. For these reasons, this study was designed to assess the feasibility of using a simple, low-cost method to detect indicators of fecal contamination of drinking water that could be implemented at the community level. Water samples from 358 sources of drinking water in the Léogâne flood basin were screened with a commercially available hydrogen sulfide test and a standard membrane method for the enumeration of thermotolerant coliforms. When compared with the gold standard method, the hydrogen sulfide test had a sensitivity of 65 % and a specificity of 93 %. While the sensitivity of the assay increased at higher fecal coliform concentrations, it never exceeded 88 %, even with fecal coliform concentrations greater than 100 colony-forming units per 100 ml. While its simplicity makes the hydrogen sulfide test attractive for assessing water quality in low-resource settings, the low sensitivity raises concerns about its use as the sole indicator of the presence or absence of fecal coliforms in individual or community water sources.     

Faecal Coliform Bacteria in Febros River (Northwest Portugal): Temporal Variation, Correlation with Water Parameters, and Species Identification

Febros river water was sampled weekly, during 35 successive weeks, and analyzed for microbiological (total coliforms, faecal coliforms, faecal streptococci and enterococci) and chemical-physical (ammonia and temperature) parameters. All microbiological parameters were highly correlated with each other and with ammonia, suggesting that the simultaneous determination of all variables currently in use in the evaluation of the microbiological quality of waters is probably redundant, and could be simplified, and that ammonia should be tested as a sentinel parameter of the microbiological pollution load of Febros river. From the strains isolated from positive tubes of the faecal coliforms test (multiple tube fermentation technique) and retested in this assay, Escherichia coli, Klebsiella oxytoca and Klebsiella pneumoniae subsp. pneumoniae strains were positive, indicating that the faecal coliforms test is not totally specific for Escherichia coli, and can detect other bacteria. Considering that these Klebsiella spp. are not necessarily of faecal origin, it was concluded that the faecal coliforms test can overestimate true faecal pollution. From the strains isolated from positive tubes of the faecal coliforms procedure, only Escherichia coli strains were clearly positive in the β-D-glucuronidase test. All other species were negative or very weakly positive, suggesting that the assay of the β-D-glucuronidase activity is less prone to false positives than the faecal coliforms test in the quantification of Escherichia coli in environmental waters.     

Analysis of the usefulness of biological parameters for the control of activated sludge wastewater treatment plants in an interlaboratory study context

The quality of the sludge in Wastewater Treatment Plants (WWTPs) depends on the suitable colonization of the flocs by microorganisms. Due to the functional importance of these biological constituents, several biological or biological-related parameters have been commonly used for the control of depuration efficiency. According to national and international water regulation recommendations, interlaboratory studies have a great relevance to determine which parameters are more reliable for their extensive application in routine control. However, these studies are also very useful to demonstrate consistency in results from multiple laboratories and to develop reliable and reproducible methodologies which might be necessary for protocol validation and also for accreditation issues to meet regulatory environmental requirements. The main purpose of this work was to assess the results obtained in consecutive interlaboratory assays in order to determine the concordance degree in the application of biological parameters by participating laboratories. Following the international recommendations about these studies, a common working protocol was proposed. Statistical tests indicated that Sludge Index and several routine physical-chemical analyses [V30, Mixed Liquor Suspended Solids (MLSS), Mixed Liquor Volatile Solids (MLVS) and Sludge Volumetric Index (SVI)] show low variability and therefore are suitable tools for laboratory control. Shannon Index and Sludge Biotic Index also presented low variability although a more precise protocol would be necessary, in particular the methodology to count small flagellates. The abundance and identification of protist species showed low concordance among laboratories and three factors were responsible for the low reliability of data: population density, size and morphological distinguishable characters of the specimens.     

Strategies to optimize monitoring schemes of recreational waters from Salta,Argentina: a multivariate approach

Several recreational surface waters in Salta, Argentina, were selected to assess their quality. Seventy percent of the measurements exceeded at least one of the limits established by international legislation becoming unsuitable for their use. To interpret results of complex data, multivariate techniques were applied. Arenales River, due to the variability observed in the data, was divided in two: upstream and downstream representing low and high pollution sites, respectively, and cluster analysis supported that differentiation. Arenales River downstream and Campo Alegre Reservoir were the most different environments, and Vaqueros and La Caldera rivers were the most similar. Canonical correlation analysis allowed exploration of correlations between physicochemical and microbiological variables except in both parts of Arenales River, and principal component analysis allowed finding relationships among the nine measured variables in all aquatic environments. Variable’s loadings showed that Arenales River downstream was impacted by industrial and domestic activities, Arenales River upstream was affected by agricultural activities, Campo Alegre Reservoir was disturbed by anthropogenic and ecological effects, and La Caldera and Vaqueros rivers were influenced by recreational activities. Discriminant analysis allowed identification of subgroup of variables responsible for seasonal and spatial variations. Enterococcus, dissolved oxygen, conductivity, E. coli, pH, and fecal coliforms are sufficient to spatially describe the quality of the aquatic environments. Regarding seasonal variations, dissolved oxygen, conductivity, fecal coliforms, and pH can be used to describe water quality during dry season, while dissolved oxygen, conductivity, total coliforms, E. coli, and Enterococcus during wet season. Thus, the use of multivariate techniques allowed optimizing monitoring tasks and minimizing costs involved.     

Comparison of fecal coliform bacteria before and after wastewater treatment plant in the Izmir Bay (Eastern Aegean Sea)

The distribution of fecal coliforms was investigated and determined in Izmir Bay from 1996 to 2005. Izmir Bay severely was polluted from industrial and domestic discharges during decades. In early 2000, a wastewater treatment plant began to treat domestic and industrial wastes. This plant treats the wastes about 80% capacity after 2001. The sampling periods cover before and after treatment plant. Assessment method for determining the number of fecal coliform has evolved membrane filtrations. Maximum surface fecal coliform concentration was 4.9 × 10⁵ cfu 100 ml^???1 in 1996–2000 period. Following the opening treatment system, fecal coliform density decreased 2.1 × 10⁴ cfu 100 ml^???1 during 2001–2005. A continuous improvement can be sustained in the water quality if direct inflow of untreated wastewater is prevented.     

粪大肠菌群酶底物法在环境应急监测中的应用

结合国外粪大肠菌群的酶底物检测方法，针对某次突发环境污染事件，用酶底物法和标准方法多管发酵法同步检测受污染地表水中的粪大肠菌群，讨论酶底物法在应急监测中检测粪大肠菌群的适用性。结果表明，两种方法的测定数据显著相关，没有统计学差异（ P＞0．05）。相对于多管发酵法，酶底物法特异性强，检测时间短，二次污染少，符合应急监测的要求。     

Intensive water quality monitoring in a Taiwan bathing beach

A total of 357 water samples were collected from a public beach in northern Taiwan during beach season, and the densities of enterococci were analyzed by Enterolert methods. The mean enterococci level was 356 MPN/100 ml and ranged from <10 to 2,005 MPN/100 ml, which was classified as high contamination level according to the WHO water quality guideline (95 percentile >1,000 MPN/100 ml). Most of the deteriorated water quality conditions occurred during rainfall. By excluding data from the rain days, the overall beach water quality would be considered in the moderate contamination level (95 percentile 200-1,000 MPN/100 ml). Among the selected microbiological parameters tested, the densities of total coliforms and enterococci exhibited the highest correlation (r = 0.449, p = 0.009), followed by the concentrations of total coliforms and fecal coliforms (r = 0.403, p = 0.02). Nonetheless, no significant correlation was found between enterococci and fecal coliform levels (r = 0.197, p = 0.271).     

Bacteriological safety assessment,hygienic habits and cross-contamination risks in a Nigerian urban sample of household kitchen environment

Urban household kitchen environment was assessed for safety by determining their levels of indicator bacteria, hygienic habits and risk of cross-contamination. Household kitchens (60) were selected in Warri Town, Nigeria, by the multi-stage sampling technique. Contact surfaces, water and indoor kitchen air were analysed for aerobic plate counts, total and faecal coliforms using Nutrient and McConkey media by swab/rinse method, membrane filtration and sedimentation methods, respectively. Hygienic habits and risk of cross-contamination were assessed with structured questionnaire which included socio-demographic variables. On the basis of median counts, the prevalence of high counts (log cfu/cm²/m³/100 mL) of aerobic plate counts (>3.0), total coliforms (>1.0) and faecal coliforms (>0) on contact surfaces and air was high (58.0–92.0%), but low in water (30.0–40.0%). Pots, plates and cutleries were the contact surfaces with low counts. Prevalence of poor hygienic habits and high risk of cross-contamination was 38.6 and 67.5%, respectively. Education, occupation and kitchen type were associated with cross-contamination risk (P = 0.002–0.022), while only education was associated with hygienic habits (P = 0.03). Cross-contamination risk was related (P = 0.01–0.05) to aerobic plate counts (OR 2.30; CL 1.30–3.17), total coliforms (OR 5.63; CL 2.76–8.25) and faecal coliforms (OR 4.24; CL 2.87–6.24), while hygienic habit was not. It can be concluded that urban household kitchens in the Nigerian setting are vulnerable to pathogens likely to cause food-borne infections.     

Applicability of universal Bacteroidales genetic marker for microbial monitoring of drinking water sources in comparison to conventional indicators

Water quality monitoring is essential for the provision of safe drinking water. In this study, we compared a selection of fecal indicators with universal Bacteroidales genetic marker to identify fecal pollution of a variety of drinking water sources. A total of 60 samples were collected from water sources. The microbiological parameters included total coliforms, fecal coliforms, Escherichia coli and fecal streptococci as the fecal indicator bacteria (FIB), Clostridium perfringens and H₂S bacteria as alternative indicators, universal Bacteroidales genetic marker as a promising alternative fecal indicator, and Salmonella spp., Shigella spp., and E. coli O157 as pathogenic bacteria. From 60 samples analyzed, Bacteroidales was the most frequently detected indicator followed by total coliforms. However, the Bacteroidales assay failed to detect the marker in nine samples positive for FIB and other alternative indicators. The results of our study showed that the absence of Bacteroidales is not necessarily an evidence of fecal and pathogenic bacteria absence and may be unable to ensure the safety of the water. Further research, however, is required for a better understanding of the use of a Bacteroidales genetic marker as an indicator in water quality monitoring programs.     

藻密度手工监测实验室间比对研究

为研究不同实验室藻密度监测数据的可靠性,现场采集了藻密度样品,处理为现场平行样和实验室前处理后平行样2种,分别由5家实验室的6名技术人员开展检测分析。参考相关技术规范要求,结合其他领域相似工作的成功应用案例,采用Z比分数法对分析结果开展藻密度实验室间比对。结果表明,数据的相对偏差均符合技术规范要求;对藻密度进行数据直接比对和对数转换后比对,各实验室对现场平行样和实验室前处理后平行样的检测结果均为合格;分析方法原理导致藻密度手工监测的绝对数值偏差较大,对当前的太湖水华预警工作适用性不高。     

水质标准中指示微生物的发展及现状

饮用水病原微生物污染是公共卫生面临的主要威胁之一,微生物监测在水质监测中的必要性日益受到人们的认可。在实际工作中,一般是通过检测指示微生物间接反映病原微生物的存在。通过调研国内外各组织、机构颁布的水质标准发现,近年来,我国环境质量标准和污染物排放标准中对于指示微生物的选择有从总大肠菌群向粪大肠菌群和大肠埃希氏菌(E.coli)转变的趋势,而美国环保署、欧盟、世界卫生组织、澳大利亚国家健康与医疗研究委员会等根据最新的流行病学证据,强调了大肠埃希氏菌(E.coli)、肠球菌(Enterococci)与粪便污染的相关性更强,可用于替代大肠菌群。建议我国在后续水质标准中对微生物指标进行增补修订时,参考国外经验,形成集成多种指示微生物与多种特定病原体的监测指标体系,以更好地保护环境功能和民众健康。