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.     

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。     

Monitoring E. coli and total coliforms in natural spring water as related to recreational mountain areas

Natural spring water has unique properties, as it is rich in minerals that are considered to be beneficial to human health. A survey of the microbiological quality of natural spring water was conducted to assess possible risks from the consumption of the water by visitors in recreational mountain areas located in Seoul, South Korea. The densities of total coliforms and Escherichia coli were measured during the spring and the summer of 2002 to investigate the presence of coliform bacteria in the drinking spring waters. Total coliforms were detected in all samples and the mean density of total coliforms was up to a maximum of 228 CFU/mL. Detectable E. coli was found in 78% of all samples and the mean densities of E. coli varied from a minimum of 0 CFU/mL to a maximum of 15 CFU/mL in all samples. Malfunctioning septic systems and wildlife population appear to be the main source of E. coli contamination. Presence of E. coli in natural spring water indicates potential adverse health effects for individuals or populations exposed to this water. The fecal contaminated spring water may present an unacceptable risk to humans if it is used as raw drinking water.     

Comparative study of fluorogenic and chromogenic media for specific detection of environmental isolates of thermotolerant Escherichia coli

In a field study 78 water samples were analysed employingFluorocult Brilla Broth (BB) and its performance was comparedwith standard MPN procedure. Out of 78 water samples analysed 56(71.7%) samples yielded positive reactions in BB whereas, 50(64.1%) samples were positive by standard fecal coliform test.A comparative study of fluorogenic and chromogenic mediacontaining substrate -D glucuronide for specificdetection of environmental isolates of 313 thermotolerant E.coli has been undertaken. Five fluorogenic media wereused: Fluorocult MacConkey agar (MCA), Fluorocult ECD agar(ECD), Fluorocult VRB agar (VRB), Fluorocult E. coli0157:H7 agar (ECH7) and Fluorocult Brilla Broth (BB) andChromogenic Chromocult agar (CCA). BB and CCA were found to behighly specific and sensitive media to detect E. coli asall E. coli yielded positive reaction on them. On ECH7 andECD agar 67.5 and 64.9 of E. coli isolates gave positivereaction, respectively. Low sensitivity was observed in case ofMCA and VRB agar in detecting E. coli. The performance ofBB appears to be better when compared with standard MPNprocedure employing MacConkey broth/Brilliant green bile brothin detecting E. coli in drinking water.     

Characterization and statistical modeling of bacterial (Escherichia coli) outflows from watersheds that discharge into southern Lake Michigan

Two watersheds in northwestern Indiana were selected for detailed monitoring of bacterially contaminated discharges (Escherichia coli) into Lake Michigan. A large watershed that drains an urbanized area with treatment plants that release raw sewage during storms discharges into Lake Michigan at the outlet of Burns Ditch. A small watershed drains part of the Great Marsh, a wetland complex that has been disrupted by ditching and limited residential development, at the outlet of Derby Ditch. Monitoring at the outlet of Burns Ditch in 1999 and 2000 indicated that E. coli concentrations vary over two orders of magnitude during storms. During one storm, sewage overflows caused concentrations to increase to more than 10,000 cfu/100 mL for several hours. Monitoring at Derby Ditch from 1997 to 2000 also indicated that E. coli concentrations increase during storms with the highest concentrations generally occurring during rising streamflow. Multiple regression analysis indicated that 60% of the variability in measured outflows of E. coli from Derby Ditch (n = 88) could be accounted for by a model that utilizes continuously measured rainfall, stream discharge, soil temperature and depth to water table in the Great Marsh. A similar analysis indicated that 90% of the variability in measured E. coli concentrations at the outlet of Burns Ditch (n = 43) during storms could be accounted for by a combination of continuously measured water-quality variables including nitrate and ammonium. These models, which utilize data that can be collected on a real-time basis, could form part of an Early Warning System for predicting beach closures.     

Linking on-farm dairy management practices to storm-flow fecal coliform loading for California coastal watersheds

How and where to improve water quality within an agricultural watershed requires data at a spatial scale that corresponds with individual management decision units on an agricultural operation. This is particularly true in the context of water quality regulations, such as Total Maximum Daily Loads (TMDLs), that identify agriculture as one source of non-point source pollution through larger tributary watershed scale and above and below water quality investigations. We have conducted a systems approach study of 10 coastal dairies and ranches to document fecal coliform concentration and loading to surface waters at the management decision unit scale. Water quality samples were collected on a storm event basis from loading units that included: manure management systems; gutters; storm drains; pastures; and corrals and lots. In addition, in-stream samples were collected above and below the dairy facilities and from a control watershed, managed for light grazing and without a dairy facility or human residence and corresponding septic system. Samples were analyzed for fecal coliform concentration by membrane filtration. Instantaneous discharge was measured for each collected sample. Storm runoff was also calculated using the curve number method (SCS, 1985). Results for a representative dairy as well as the entire 10 dairy data set are presented. Fecal coliform concentrations demonstrate high variability both within and between loading units. Fecal coliform concentrations for pastures range from 206 to 2,288,888 cfu/100 ml and for lots from 1,933 to 166,105,000 cfu/100 ml. Mean concentrations for pastures and lots are 121,298 (SE=62,222) and 3,155,584 (SE=1,902,713) cfu/100 ml, respectively. Fecal coliform load from units of concentrated animals and manure are significantly more than units such as pastures while storm flow amounts were significantly less. Compared with results from earlier tributary scale studies in the watershed, this systems approach has generatedwater quality data that is beneficial for management decisions because of its scale and representation of current management activities. These results are facilitating on-farm changes through the cooperative efforts of dairy managers, regulatory agency staff, and sources of technical and financial assistance.     

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.     

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.     

Characterization of Escherichia Coli Isolates from Different Fecal Sources by Means of Classification Tree Analysis of Fatty Acid Methyl Ester (Fame) Profiles

Microbial source tracking (MST) methods need to be rapid, inexpensive and accurate. Unfortunately, many MST methods provide a wealth of information that is difficult to interpret by the regulators who use this information to make decisions. This paper describes the use of classification tree analysis to interpret the results of a MST method based on fatty acid methyl ester (FAME) profiles of Escherichia coli isolates, and to present results in a format readily interpretable by water quality managers. Raw sewage E. coli isolates and animal E. coli isolates from cow, dog, gull, and horse were isolated and their FAME profiles collected. Correct classification rates determined with leaveone-out cross-validation resulted in an overall low correct classification rate of 61%. A higher overall correct classification rate of 85% was obtained when the animal isolates were pooled together and compared to the raw sewage isolates. Bootstrap aggregation or adaptive resampling and combining of the FAME profile data increased correct classification rates substantially. Other MST methods may be better suited to differentiate between different fecal sources but classification tree analysis has enabled us to distinguish raw sewage from animal E. coli isolates, which previously had not been possible with other multivariate methods such as principal component analysis and cluster analysis.     

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.