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.     

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。     

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。     

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。     

纸片法快速测定水质总大肠菌群

简要介绍了纸片法测定水质总大肠菌群的操作方法，并通过６个实验室的验证试验，研究了纸片法与现行标准方法（多管法）的一致性。结果表明，２种方法的测定结果基本一致，无显著性差异，但纸片法测定周期比多管法缩短了２ｄ，仅需１ｄ即可。     

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。     

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.     

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-…     

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。     

不同污染指示菌对河流的细菌学评价

以细菌学理论为依据，以湖南沅江常德江段水体为实例，说明了用《地表水环境质量标准》（GHZB 1－1999）中粪大肠菌群指标（FC）和《地面水环境质量标准》（GB 3838－88）中总大肠菌群指标（TC）分别评价河流水体的细菌学质量所存在的差异。     

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.     

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.     

Sources and fate of Salmonella and fecal indicator bacteria in an urban creek

This research aimed to understand the sources and fate of Salmonella and fecal bacteria in urban surface waters. An urban creek (San Pedro Creek, California, USA) that had unusually high levels of Salmonella and fecal bacteria relative to other nearby waterbodies was chosen as a model field site. State of the art microbiological methods were used in concert with modeling to investigate Salmonella and fecal bacteria sources, and determine field-relevant dark inactivation and photoinactivation rates. Three along-creek surveys that spanned reaches adjacent to both urban and forested land covers were conducted to measure Salmonella, enterococci, Escherichia coli, and horse- and human-specific Bacteroidales. Salmonella were detected adjacent to and downstream of urban land cover, but not adjacent to forested land cover. No human or horse-specific Bacteroidales fecal markers were detected implicating other urban animal sources of bacteria. Two locations along the creek where Salmonella was consistently detected were sampled hourly for 25 hours and a mass-balance model was applied to determine field-relevant light and dark inactivation rates for Salmonella, enterococci, and E. coli. Sunlight inactivation did not appear to be important in modulating concentrations of Salmonella, but was important in modulating both enterococci and E. coli concentrations. Dark inactivation was important for all three organisms. This is the first study to quantitatively examine the fate of Salmonella within an urban surface water. Although the work is carried out at a single site, the methodologies are extendable to source tracking in other waterbodies. Additionally, the rate constants determined through the modeling will be useful for modeling these organisms in other surface waters, and represent useful benchmarks for comparison to laboratory-derived inactivation rates.     

A comparison of rapid and conventional measures of indicator bacteria as predictors of waterborne protozoan pathogen presence and density

E. coli and enterococci in recreational waters are monitored as indicators of fecal contamination, pathogen presence, and health risk. Quantitative polymerase chain reaction (qPCR) tests for fecal indicator bacteria can provide beach managers with same-day information about water quality, unlike culture methods which provide that information the following day. The abilities of qPCR measurements of indicator bacteria, as compared to culture measurements of indicator bacteria, as predictors of pathogen presence or density in surface waters are not well understood. The purpose of this study was to make such comparisons between water samples collected from Chicago area surface waters, including rivers, inland lakes, Lake Michigan, and the Chicago Area Waterways System, which is dominated by wastewater effluent. A total of 294 twenty-litre samples were collected and analyzed for Giardia and Cryptosporidium. qPCR and membrane filtration methods were used to quantify E. coli and enterococci. Correlation, logistic regression, and zero-inflated Poisson modeling were utilized to evaluate associations between indicators and parasites. qPCR and culture measures of the indicator bacteria were similar in their ability to predict parasite presence and density. Correlations between parasites and indicators were generally stronger at waters not dominated by effluent. Associations between indicator density and Giarida presence were observed more consistently than between indicator density and Cryptosporidium presence. Associations between enterococci and parasites were generally stronger than associations between E. coli and parasites. The use of qPCR monitoring in our setting would generate more timely results without compromising the ability to predict parasite presence or density.     

Microbiological water quality of the Mfoundi River watershed at Yaoundé, Cameroon, as inferred from indicator bacteria of fecal contamination

Using the membrane filtration technique to count total coliform (TC), fecal coliform (FC) and fecal streptococci (FS), the microbiological water quality of the Mfoundi River and four of its representative tributaries at Yaoundé, Cameroon, was assessed for human use and contact. Sampling was conducted so as to examine the potential origin of fecal contamination and how rainfall affects the measured concentrations of indicators organisms. Our results revealed that waters were not safe for human use or primary contact according to the standards for water quality established by the Word Health Organization (WHO). Indeed, these waters exhibited high concentrations of TC (Mean ± SD=5.6 × 10⁸ ± 2.5 × 10⁶ CFU/100 ml), FC (Mean ± SD=6.8 × 10⁵ ± 2.4 × 10³ CFU/100 ml) and FS (Mean ± SD=7.3 × 10⁵ ± 2.1 × 10³ CFU/100 ml) that varied with the sampling sites and points. FC/FS ratio suggested that this contamination was more from warm-blooded animals than humans and correlation analysis points to the role of rainfall as a contributing factor, which enhanced the bacterial numbers detected. We conclude that there is a great potential risk of infection for users of waters from the Mfoundi River and its tributaries at Yaoundé.     

E. coli kinetics--effect of temperature on the maintenance and respectively the decay phase

The knowledge of enteric bacteria survival kinetic is very important for environmental scientists. Enteric bacteria andspecifically the fecal indicator bacteria are typically used to measure the sanitary quality of water for recreational, industrial, agricultural and water supply purposes. They are released into the environment with feces, and are then exposedto a variety of environmental conditions that eventually causetheir death. In general, it is believed that the fecal indicatorcannot grow in natural environments, since they are adapted to live in the gastrointestinal tract. Studies have shown that fecalindicator bacteria survive from a few hours up to several daysin surface water, but may survive for days or months in lake-sediments, where they may be protected from sunlight andpredators. We assume that pathogens similar to the fecal indicator bacteria die at the same rate as fecal indicator bacteria. Therefore, if we find relatively high numbers of fecalindicator bacteria in an environment, we assume that there is anincreased likelihood of pathogens being present as well. The kinetic of enteric bacteria survival in natural waters is affected by a large number of factors. One of them is the temperature. The aim of this contribution was the experimentalresearch of the survival kinetic of enteric bacteria applying a simple mathematical formula, which describes the survival kineticpredicting the decay phase at various temperatures. We aspire that the results will lead both to the solution of many engineering problems and to future research.     

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.     

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.     

Assessment of animal impacts on bacterial water quality in a South Carolina, USA tidal creek system

Fecal pollution may adversely impact water quality in coastal ecosystems. The goal of this study was to determine whether cattle were a source of fecal pollution in a South Carolina watershed. Surface water samples were collected in June 2002 and February through March 2003 in closed shellfish harvesting waters of Toogoodoo Creek in Charleston County, SC. Fecal coliform concentrations in 70 % of the water samples taken for this study exceeded shellfish harvesting water standards. Ribotyping was performed in order to identify animal sources contributing to elevated fecal coliform levels. Escherichia coli isolates (n?=?253) from surface water samples were ribotyped and compared to a ribotype library developed from known sources of fecal material. Ribotypes from water samples that matched library ribotypes with 90 % maximum similarity or better were assigned to that source. Less than half of the unknown isolates (38 %) matched with library isolates. About half (53 %) of the matched ribotypes were assigned to cattle isolates and 43 % to raccoon. Ribotyping almost exclusively identified animal sources. While these results indicate that runoff from cattle farms was a likely source of fecal pollution in the watershed, wildlife also contributed. Given the small size of the library, ribotyping was moderately useful for determining the impact of adjacent cattle farms on Toogoodoo Creek. Increasing the number and diversity of the wildlife sources from the area would likely increase the usefulness of the method.