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。     

关于垃圾填埋场渗滤液的大肠菌值指标的探讨

指出了监测生活垃圾填埋物渗滤液中遇到的大肠菌群检测方法和排放限值单位应用的问题，并建议使用粪大肠菌群作为指标，开展定性检测。     

水中粪大肠菌群快速检测方法-固定底物酶底物法与多管发酵法的比较

目的在于比较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)的检测,使用科立得TM(Colilert(R))试刺和传统方法检测地表水、水源水及污水水样,比较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)检测结果的一致性.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,固定底物酶底物法可以用作评价水质微生物污染的标准方法.     

两种酶底物法监测水中(粪)大肠菌群商品化制品的比较

比较了进口colilert试剂和国产colitech试剂及其各自配套制品对水中(粪)大肠菌群的检测,包括总大肠菌群和粪大肠菌群。分别使用同为酶底物法的两种制品检测地表水、地下水、水源水、生活饮用水及污水水样,比较了两种制品用于水中(粪)大肠菌群检测结果的一致性,同时验证了两种制品的无菌性和培养基的选择性。试验结果表明,进口colilert试剂和国产colitech试剂及其各自配套制品于水中(粪)大肠菌群检测结果具有一致性,培养基具有很好的选择性。     

五管发酵测定粪大肠菌群的研究

通过实验摸索出粪大肠菌群的一种五管发酵法。该方法检测下限为9个/L,适用于地表水和废水中粪大肠菌群的测定。     

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。     

快速测定地表水中粪大肠菌群

地表水中粪大肠菌群是评价水质卫生状况的一项重要指标,一般采用多管发酵标准法测定。该方法工作量大,操作繁琐,周期长。因粪大肠菌群的复发酵试验在44 5℃条件下仍能以产酸产气加以判断,因此,可直接将地表水样接种于EC培养液中,按多管发酵标准法原理计算粪大肠杆菌群数,快速检测。1　材料EC培养液,试管,小倒管等。2　检测方法2 1　操作步骤调节地表水样pH值至中性(7～8),按多管发酵标准法的稀释接种方法对水样作适当稀释,以EC培养液直接接种水样,在44 5℃水浴中培养24h,观察结果。表1　多个样品粪大肠菌群两种测定方法的结果　　L-…     

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。     

24孔最大可能数法快速检测水质粪大肠菌群

设计了一种24孔最大可能数法,用玫红酸抑制杂菌,快速检测水质粪大肠菌群。方法能在24 h内检测不同程度污染水样中103L-1~107L-1的粪大肠菌群,回收率为76.1%~108%,批内RSD为24.0%~34.0%。与多管发酵法的比较试验表明,两种方法相关性良好,且在添加玫红酸条件下等效。提出方法还需要更多不同性质及来源水样的检测结果来验证。     

延长初发酵培养时间对大肠菌群结果影响分析

阐述了将《大肠菌群多管发酵法》(GB/T 4789.28-2003)初发酵时间24 h延长至48 h的原因,通过对139份不同种类样品进行大肠菌群总数检测,发现有12份样品增加了大肠菌群数,表明延长初发酵培养时间,可以增加迟缓发酵大肠菌群成员检出.指出迟缓发酵大肠菌群在44.5 ℃培养时,均无产酸产气现象,表明引起迟缓发酵的大肠菌群不属于粪大肠菌群成员.     

Chlorination and water quality monitoring within a public drinking water supply in Rawalpindi Cantt (Westridge and Tench) area, Pakistan

Concern over the presence of fecal coliform in public drinking water supplies has been expressed in recent years in Pakistan since it has been regarded as pathogenic organism of prime importance in gastroenteritis. Two major drinking water distribution systems in the Cantt area of Rawalpindi district covering the Westridge and Tench areas was monitored over a 2-month period to determine the prevalence of fecal coliform and chlorine residual. The collected samples were examined for total chlorine, free chlorine residual, chloramines, total coliforms, fecal coliforms, and turbidity. The drinking water quality monitoring in the distribution network was performed by collecting samples from water source, overhead reservoir, and residential taps. In the Westridge area, total chlorine varied from the lowest value of 0.27 mg/L at Station # W-5 to the highest value of 0.42 mg/L at Station # W-2, total coliforms varied from 1.1 to 3.6 most probable number (MPN)/100 mL with presence of Escherichia coli in all samples, total dissolved solids (TDS) ranged from 199.5 to 205 mg/L, conductivity fluctuated between 399 and 411 microS/cm, and turbidity varied from 0.43 to 0.73 NTU. In the Tench area, the value of total chlorine ranged from 0.14 mg/L at Station # T-7 to 0.55 mg/L at Station # T-1. Total coliform varied from 3.6 to 5.1 MPN/100 mL and fecal coliform were detected at all the stations except at Station # T-1. TDS ranged from 201.4 to 257 mg/L, conductivity varied from 343 to 513 microS/cm, and turbidity ranged between 0.66 and 1.55 NTU. It is recommended to the respective agencies to ensure that the chlorine residual is available at consumer end.     

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.     

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.     

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.     

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).     

Identifying fecal pollution sources using 3M™ Petrifilm™ count plates and antibiotic resistance analysis in the Horse Creek Watershed in Aiken County,SC (USA)

Sources of fecal coliform pollution in a small South Carolina (USA) watershed were identified using inexpensive methods and commonly available equipment. Samples from the upper reaches of the watershed were analyzed with 3M^? Petrifilm^? count plates. We were able to narrow down the study’s focus to one particular tributary, Sand River, that was the major contributor of the coliform pollution (both fecal and total) to a downstream reservoir that is heavily used for recreation purposes. Concentrations of total coliforms ranged from 2,400 to 120,333 cfu/100 mL, with sharp increases in coliform counts observed in samples taken after rain events. Positive correlations between turbidity and fecal coliform counts suggested a relationship between fecal pollution and stormwater runoff. Antibiotic resistance analysis (ARA) compared antibiotic resistance profiles of fecal coliform isolates from the stream to those of a watershed-specific fecal source library (equine, waterfowl, canines, and untreated sewage). Known fecal source isolates and unknown isolates from the stream were exposed to six antibiotics at three concentrations each. Discriminant analysis grouped known isolates with an overall average rate of correct classification (ARCC) of 84.3 %. A total of 401 isolates from the first stream location were classified as equine (45.9 %), sewage (39.4 %), waterfowl (6.2 %), and feline (8.5 %). A similar pattern was observed at the second sampling location, with 42.6 % equine, 45.2 % sewage, 2.8 % waterfowl, 0.6 % canine, and 8.8 % feline. While there were slight weather-dependent differences, the vast majority of the coliform pollution in this stream appeared to be from two sources, equine and sewage. This information will contribute to better land use decisions and further justify implementation of low-impact development practices within this urban watershed.     

Bacteria holding times for fecal coliform by mFC agar method and total coliform and Escherichia coli by Colilert®-18 Quanti-Tray® method

Bacteria holding-time experiments of up to 62 h were performed on five surface-water samples from four urban stream sites in the vicinity of Atlanta, GA, USA that had relatively high densities of coliform bacteria (Escherichia coli densities were all well above the US Environmental Protection Agency criterion of 126 colonies (100 ml)^???1 for recreational waters). Holding-time experiments were done for fecal coliform using the membrane filtration modified fecal coliform (mFC) agar method and for total coliform and E. coli using the Colilert^®-18 Quanti-Tray^® method. The precisions of these analytical methods were quantified. Precisions determined for fecal coliform indicated that the upper bound of the ideal range of counts could reasonably be extended upward and would improve precision. For the Colilert^®-18 method, analytical precisions were similar to the theoretical precisions for this method. Fecal and total coliform densities did not change significantly with holding times up to about 27 h. Limited information indicated that fecal coliform densities might be stable for holding times of up to 62 h, whereas total coliform densities might not be stable for holding times greater than about 27 h. E. coli densities were stable for holding times of up to 18 h—a shorter period than indicated from a previous studies. These results should be applicable to non-regulatory monitoring sampling designs for similar urban surface-water sample types.     

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.