水中粪大肠菌群检测方法及其研究进展

水体受粪便污染后可导致肠道病毒的扩散和传播，粪大肠菌群恰可准确地反映水体受粪便污染的状况，因此，对粪大肠菌群的检测是否精确、快速对疾病的预防有至关重要的作用。本文对目前粪大肠菌群的分析方法及其研究改进进行了综述和比较，认为目前水中粪大肠菌群的检测可优先选择纸片法和酶底物法，而经过改进的多管发酵法（EC培养法，乳糖蛋白胨培养法），若再加以验证完善，亦可成为一个好的选择。     

对水的细菌学检测中几个问题的讨论

“大肠菌群”是水质污染的指示菌，测定大肠菌群的方法主要有多管发酵法、滤膜法。多管发酵法实验周期较长，适用于较复杂的水样检测。本文对多管发酵法实验过程中的一些具体问题、如革兰氏染色法中的注意事项、初发酵24h内产酸不产气样品的处理，多管发酵法操作步骤是否可简化等、结合实践经验进行了讨论和分析。     

固定底物酶底物法与多管发酵法和快速纸片法测定水中粪大肠菌群(耐热大肠菌群)的比较

采用酶底物法和多管发酵法及快速纸片法,比对分析了分组环境水样中的大肠菌群,采用无菌水和标准菌种ATCC 25922(大肠埃希菌)标准菌种检验3种方法的假阳性率与假阴性率,结果表明,固定底物酶底物法与多管发酵法用于水中粪大肠菌群(耐热大肠菌群)检测结果具有一致性。     

水中粪大肠菌群结果不确定度的评定

依据国家标准和测量不确定度的评定方法,评定水中粪大肠菌群的不确定度.认为不确定度的主要来源为样品保存条件、培养条件、人员操作和判定计数等.通过对地表水和海水样品粪大肠菌群操作过程中的不确定度进行分量计算,建立一种简单的水中粪大肠菌群多管发酵法不确定度评定方法.     

关于饮水机水细菌污染及使用若干问题探讨

为了解冷热两用饮水机所供水的细菌污染情况，对新出厂的饮用纯净水、饮水机中冷水、未沸水和沸水、桶中水等不同水样中细菌总数和大肠菌群分别进行了检测分析。结果表明，未沸水灭菌情况不稳定，沸水灭菌效果很好；随时间推移，纯净水中的细菌总数呈递增趋势，并且增长速度较快，其污染源主要来自空气和饮水机本身，特别是饮水机内胆的污染不容忽视。所有水样中均未检出大肠菌群。     

宁波市主要流域农用水质量调查与评价

通过对宁波市两大主要流域姚江、奉化江农用水质环境质量的监测和综合分析评价,结果表明：两流域上游水质较好,没有监测指标超标,达到1级水质标准;中下游水质较差,有多项指标超标,水质以3级为主,部分地段水质达到2级,其主要超标物为粪大肠菌群数、悬浮物、石油类、六价铬和化学需氧量等,其中粪大肠菌群数、悬浮物是影响两流域水体质量的主要污染因子。姚江流域水质要明显好于奉化江流域。在水质的具体农业应用上应先考虑粮油作物和其他旱作作物的灌溉,若使用在对水质要求较严的蔬菜灌溉或渔业养殖上,建议对水质进行治理后再应用,以防影响农产品的产量和品质。     

武夷山九曲溪水环境质量综合评价

根据武夷山九曲溪2003-2007年水质监测资料,用均值型综合污染指数法对九曲溪水环境进行评价.结果表明,2003-2007年九曲溪水环境污染程度均为尚清洁级别,各断面水环境污染程度由重到轻依次为:星村码头>江墩桥>漫水桥>玉女峰>曹墩桥;溪流的主要污染物是粪大肠菌群,其次是五日生化需氧量、高锰酸盐指数和溶解氧;污染源主要是沿岸生活垃圾渗滤液和一些粪便污水,竹筏漂流对水质影响很小.     

密封消解法测定高氯离子含盐废水COD_(Cr)的探讨

针对国标重铬酸钾法测定高氯离子含盐废水CODCr时的不足,提出用密封消解法来测定高盐废水CODCr的观点,通过丁酮氧化率、氯离子干扰、混配水样和实际水样测定结果的比较,对国标法和密封消解法进行了验证。试验结果表明:在测定高氯离子含盐废水CODCr值时,密封消解法优于重铬酸钾法,能够真实准确地反映废水的CODCr。     

仪器法测定水环境高锰酸盐指数的应用

高锰酸盐指数是表征水质污染程度的重要指标之一,按照GB 11892—1989《水质 高锰酸盐指数的测定》中的水浴温度、水浴时间、滴定终点颜色判断、滴定条件的要求,人为因素和实验环境因素对 高锰酸盐指数检测数据的准确性影响较大,传统手工容量法检测的弊端日益显现,因此按照环境监测标 准设计自动化检验分析设备,提高实验室检测水平尤为重要。文章对水质高锰酸盐指数全自动分析仪方 法的准确度和精密度进行了测定分析,并与国标容量法的操作过程和数据进行对比探讨,仪器法测定下限为0.05 mg/L,而国标容量法规定的测定下限为0.5 mg/L,仪器法在实际样品测定中的应用范围更 广,数据更可靠。     

涪江流域遂宁段生态环境监测与水质污染现状评价

为研究涪江遂宁段河流水环境质量和污染现状,对水体中基本理化指标和细菌总数、粪大肠茵群数、浮游藻类组成等生物指标进行了监测和综合分析,并采用了单因子评价法和综合污染指数法对其进行水质污染现状进行了评价。结果表明,该流域境内香山和郭江属地表水Ⅳ类中度污染,污染指标总氮、总磷和BOD;超标倍数分别为0．76、0．26、0．02和0．39、0．24、0．03;跑马滩属地表水Ⅳ类中度污染,总氮和BOD,超标倍数为0．08、0．13;米家桥和老池属于Ⅲ类轻度污染;总氮、总磷和BOD,断面超标率分别为60％、40％、60％。此外,各监测断面水体中细菌总数、大肠菌群数及藻类组成均差异明显,变化趋势与水质理化指标基本吻合。细菌总数变化趋势为郭江〉跑马滩〉香山〉米家桥〉老池,郭江细菌总数2．36×10^7cfu／mL,比跑马滩、香山高、米家桥和老池分别高23.7％和41．5％．56．25倍和65．5倍。     

Fecal coliform loadings and stocks in buttermilk bay,Massachusetts, USA,and management implications

Abundance of fecal caliform bacteria is a weak index of the presence of human pathogens in wastewater entering coastal waters. In spite of this, use of fecal caliform indices for management purposes is widespread. To gain insight into interpretation of fecal coliform data, we evaluated size of stocks of fecal coliforms in water, sediments, and sea wrack, in Buttermilk Bay, a coastal embayment in Massachusetts. Sediments contained most of the fecal coliforms. Fecal coliforms in sediments were as much as one order of magnitude more abundant than in the water column or in sea wrack. The fecal coliforms in sediments of Buttermilk Bay were so abundant that resuspension of fecal coliforms from just the top 2 cm of muddy sediments could add sufficient cells to the water column to have the whole bay exceed the federal limit of fecal coliforms for shellfishing. The major sources of fecal coliforms to the bay were water-fowls, surface runoff, groundwater, and streams. Waterfowl were the largest source of fecal coliforms during cold months; surface runoff, streams, and groundwater were most important during warm months. Redirection of surface runoff pipes is unlikely to be a very successful management action since contributions via this source are insufficient to account for the measured increases in concentrations of fecal coliforms in water. Removal of waterfowl is also unlikely to be useful, since fecal coliform concentrations leading to closures of shellfish beds and swimming areas are most frequent during warm months when waterfowl are rarest. Rates of loss of fecal caliform cells from the water column by death and tidal exchange were high. Mortality of cells was about an order of magnitude larger than losses by tidal exchange. The amounts of fecal coliforms brought into the bay by waterfowl, surface runoff, groundwater, and streams are an order of magnitude smaller than the losses by mortality and tidal removal. This implies that there is an additional source of fecal coliforms within the bay. We suggest that resuspension of the upper layers of sediments can easily account for the fecal coliforms present in the water. Fecal coliform content of water and shellfish were not correlated. In contrast, sediment and shellfish fecal coliform abundances were significantly related. Monitoring of fecal coliforms in sediments may provide a better assessment of shellfish than sampling of water. The large fecal coliform stock in sediments should be the first priority for management. Efforts ought to be directed toward the reduction of sediment fecal coliform stocks. Lowering nutrient additions to coastal water bodies may be one practical approach.     

AN ANALYSIS OF COLIFORM BACTERIA IN THE UPPER ILLINOIS WATERWAY1

ABSTRACT: During the summer of 1971 about 150 water samples were examined for total and fecal coliform bacteria in the Upper Illinois Waterway at 19 river stations. The data per station were found to be normal geometric distributed. Bacteria densities changed with sampling dates and generally decreased with water movement downstream. Several sewage treatment effluents made marked pulses along the bacterial die-off curves. The observed fecal coliform results were evaluated in terms of the Illinois Pollution Control Board's standards. The FC:TC ratio on the waterway for each station were presented. Using Chick's Law, coliforms death rates were estimated. Efforts to correlate observed total and fecal densities with temperature, flow, algal densities, dissolved oxygen, and 5-day biochemical oxygen demand were not successful. (KEY TERMS: algae; biochemical oxygen demand; coliform bacteria; dissolved oxygen; flow; stream survey; temperature; water pollution; water quality standards)     

BACTERIAL LEVELS IN CISTERN WATER SYSTEMS OF NORTHERN KENTUCKY1

ABSTRACT: In rural Northern Kentucky, rainwater is commonly collected from rooftops and stored in cement block cisterns as the sole source of drinking water. Although every cistern system is unique in some aspect of design, use, or maintenance, a bacterial survey of 30 rural Northern Kentucky cistern systems suggests that coliforms and heterotrophic bacteria are common to all types of cistern storage systems. An average of 600 coliforms/ml and 3.6 ± 10⁵ heterotrophic bacteria/ml were detected in water samples from the bottoms of the cistern storage tanks. Bacterial levels in water delivered to household cold tap faucets were similar to the levels found in the storage tanks. When detected, fecal coliforms were recovered throughout the entire system including the household cold tap faucet. Current U.S. regulations for drinking water quality are discussed, with a suggestion that fecal coliform levels may be a more appropriate guideline for interpreting the water quality of individually maintained, nonchlorinated, nonpiped water supplies, such as cistern storage systems.     

Survival potential of Escherichia coli and Enterococci in subtropical beach sand: implications for water quality managers

Fecal bacteria have traditionally been used as indicator organisms to monitor the quality of recreational waters. Recent work has questioned the robustness of traditional indicators, particularly at seawater bathing beaches. For example, a study of Florida beaches found unexpectedly high abundances of Escherichia coli, fecal coliforms, and enterococci in beach sand. The aim of the present study was to explain these abundances by assessing the survival of E. coli and enterococci in beach sand relative to seawater. We used a combination of quantitative laboratory mesocosm experiments and field observations. Results suggested that E. coli and enterococci exhibited increased survivability and growth in sand relative to seawater. Because fecal bacteria are capable of replicating in sand, at least under controlled laboratory conditions, the results suggest that sand may be an important reservoir of metabolically active fecal organisms. Experiments with "natural" mesocosms (i.e., unsterilized sand or water rich in micropredators and native bacteria) failed to show the same increases in fecal indicators as was found in sterile sand. It is postulated that this was due to predation and competition with indigenous bacteria in these "natural" systems. Nonetheless, high populations of indicators were maintained and recovered from sand over the duration of the experiment as opposed to the die-off noted in water. Indicator bacteria may wash out of sand into shoreline waters during weather and tidal events, thereby decreasing the effectiveness of these indicators as predictors of health risk and complicating the interpretations for water quality managers.     

Evaluation of the host-specificity and prevalence of enterococci surface protein (esp) marker in sewage and its application for sourcing human fecal pollution

The suitability of the enterococci surface protein (esp) marker to detect human fecal pollution was evaluated by testing 197 fecal samples from 13 host groups in Southeast Queensland, Australia. Overall, this marker was detected in 90.5% of sewage and septic system samples and could not be detected in any fecal samples from 12 animal host groups. The sensitivity of the esp primer to detect the human-specific esp marker in sewage and septic samples was 100 and 67%, respectively. The overall specificity of this marker to distinguish between human and animal fecal pollution was 100%. Its prevalence in sewage was also determined by testing samples from the raw sewage, secondary effluent, and treated effluent of a sewage treatment plant (STP) over five consecutive days. Of the 15 samples tested, 12 (80%) were found to be positive for this marker. In contrast, it was not found in three samples from the treated effluent and these samples did not contain any culturable enterococci. The PCR limit of detection of this marker in freshwater samples was up to dilution 1 x 10(-4) and the number of culturable enterococci at this dilution was 4.8 x 10(1) +/- 7.0 x 10 degrees colony forming unit (CFU). The utility of this marker was evaluated by testing water samples from three non-sewered catchments in Pine Rivers in Southeast Queensland. Of the 13 samples tested, eight were positive for this marker with the number of enterococci ranging between 1.8 x 10(3) to 8.5 x 10(3) CFU per 100 mL of water. Based on the results, it can be concluded that the esp marker appears to be sewage specific and could be used as a reliable marker to detect human fecal pollution in surface waters in Southeast Queensland, Australia.     

Transport of fecal bacteria by boots and vehicle tires in a rural Alaskan community

People living without piped water and sewer can be at increased risk for diseases transmitted via the fecal-oral route. One rural Alaskan community that relies on hauling water into homes and sewage from homes was studied to determine the pathways of fecal contamination of drinking water and the human environment so that barriers can be established to protect health. Samples were tested for the fecal indicator, Escherichia coli, and the less specific indicator group, total coliforms. Shoes transported fecal contamination from outside to floor material inside buildings. Contamination in puddles on the road, in conjunction with contamination found on all-terrain vehicle (ATV) tires, supports vehicle traffic as a mechanism for transporting contamination from the dumpsite or other source areas to the rest of the community. The abundance of fecal bacteria transported around the community on shoes and ATV tires suggests that centralized measures for waste disposal as well as shoe removal in buildings could improve sanitation and health in the community.     

Modeling manure-borne bromide and fecal coliform transport with runoff and infiltration at a hillslope

Hillslope vegetated buffers are recommended to prevent water pollution from agricultural runoff. However, models to predict the efficacy of different grass buffer designs are lacking. The objective of this work was to develop and test a mechanistic model of coupled surface and subsurface flow and transport of bacteria and a conservative tracer on hillslopes. The testing should indicate what level of complexity and observation density might be needed to capture essential processes in the model. We combined the three-dimensional FEMWATER model of saturated-unsaturated subsurface flow with the Saint-Venant model for runoff. The model was tested with data on rainfall-induced fecal coliforms (FC) and bromide (Br) transport from manure applied at vegetated and bare 6-m long plots. The calibration of water retention parameters was unnecessary, and the same manure release parameters could be used both for simulations of Br and FC. Surface straining rates were similar for Br and bacteria. Simulations of Br and FC concentrations were least successful for the funnels closest to the source. This could be related to the finger-like flow of the manure from the strip along the bare slopes, to the transport of Br and FC with manure colloids that became strained at the grass slope, and to the presence of micro-ponds at the grassed slope. The two-dimensional model abstraction of the actual 3D transport worked well for flux-averaged concentrations. The model developed in this work is suitable to simulate surface and subsurface transport of agricultural contaminants on hillslopes and to evaluate efficiency of grass strip buffers, especially when lateral subsurface flow is important.     

Evaluating sewage-associated JCV and BKV polyomaviruses for sourcing human fecal pollution in a coastal river in Southeast Queensland, Australia

In this study, the host-sensitivity and host-specificity of JC virus (JCV) and BK virus (BKV) polyomaviruses were evaluated by testing wastewater and fecal samples from nine host groups in Southeast Queensland, Australia. The JCV and BKV polyomaviruses were detected in 63 human wastewater samples collected from primary and secondary effluent, suggesting high sensitivity of these viruses in human wastewater. In the 81 animal wastewater and fecal samples tested, 80 were polymerase chain reaction (PCR) negative for the JCV and BKV markers. Only one sample (out of 81 animal wastewater and fecal samples) from pig wastewater was positive. Nonetheless, the overall host-specificity of these viruses to differentiate between human and animal wastewater and fecal samples was 0.99. To our knowledge, this is the first study in Australia that reports on the high specificity of JCV and BKV polyomaviruses. To evaluate the field application of these viral markers for detecting human fecal pollution, 20 environmental samples were collected from a coastal river. In the 20 samples tested, 15% (3/20) and 70% (14/20) samples exceeded the regulatory guidelines for Escherichia coli and enterococci levels for marine waters. In all, five (25%) samples were PCR positive for JCV and BKV, indicating the presence of human fecal pollution in the coastal river investigated. The results suggest that JCV and BKV detection using PCR could be a useful tool for identifying human-sourced fecal pollution in coastal waters.     

Microbial and chemical markers: runoff transfer in animal manure-amended soils

Fecal contamination of water resources is evaluated by the enumeration of the fecal coliforms and Enterococci. However, the enumeration of these indicators does not allow us to differentiate between the sources of fecal contamination. Therefore, it is important to use alternative indicators of fecal contamination to identify livestock contamination in surface waters. The concentration of fecal indicators (, enteroccoci, and F-specific bacteriophages), microbiological markers (Rum-2-bac, Pig-2-bac, and ), and chemical fingerprints (sterols and stanols and other chemical compounds analyzed by 3D-fluorescence excitation-matrix spectroscopy) were determined in runoff waters generated by an artificial rainfall simulator. Three replicate plot experiments were conducted with swine slurry and cattle manure at agronomic nitrogen application rates. Low amounts of bacterial indicators (1.9-4.7%) are released in runoff water from swine-slurry-amended soils, whereas greater amounts (1.1-28.3%) of these indicators are released in runoff water from cattle-manure-amended soils. Microbial and chemical markers from animal manure were transferred to runoff water, allowing discrimination between swine and cattle fecal contamination in the environment via runoff after manure spreading. Host-specific bacterial and chemical markers were quantified for the first time in runoff waters samples after the experimental spreading of swine slurry or cattle manure.