ACCOMMODATING CHANGE OF BACTERIAL INDICATORS IN LONG TERM WATER QUALITY DATASETS1

ABSTRACT: In 1996, the State of Oregon adopted a water quality standard based on Escherichia coli (E. coli), recognizing E. coli as an indicator of pathogenic potential. The Oregon Department of Environmental Quality (DEQ) began analysis for E. coli that same year. The Oregon DEQ continued collection and analysis of fecal coliform (a prior indicator organism) for data input to bacterial loading models and the Oregon Water Quality Index (OWQI). The OWQI is a primary indicator of general water quality for the Oregon DEQ and the Oregon Progress Board. The objective of this study was to develop a regression relationship between fecal coliform and E. coli. This relationship would fill data gaps and extend water quality models and indicators. Water quality policy is better informed by the ability of these extended water quality models to determine whether water quality meets present or would have met past bacterial standards. Monitoring resources spent on dual bacterial analyses could be conserved. This study also showed that changes to OWQI values (as a result of changing bacterial indicators) were minimal, and corresponded to improved characterization of water quality with respect to pathogenic potential.     

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 commercial odor control agents for suppressing Escherichia coli in swine manure slurry

Ten commercially available manure odor control agents were evaluated in bench-scale laboratory microcosms for their ability to inhibit or kill Escherichia coli, a commonly used indicator of fecal pollution and a potential pathogen. At manufacturer recommended rates, none of the agents reduced viable populations of E. coli in pure cultures or in swine manure slurry. However, at rates 10-fold higher than those recommended by the manufacturer, EnviroPur rapidly reduced viable populations of E. coli. Accelerated death of E. coli was observed at temperatures as low as 4 degrees C. Chemical analysis of EnviroPur indicated that it contains alkylphenol polyethoxylates, common industrial surfactants. These results suggest that at manufacturer-recommended rates, the odor-controlling agents would not be effective at suppressing E. coli in stored swine manure slurry.     

Spatial and temporal modeling of microbial contaminants on grazing farmlands

This paper introduces an integrated spatial and temporal modeling system developed mathematically for assessing microbial contaminants on animal-grazed farmlands. The model uses fecal coliform, specifically Escherichia coli, as an indicator of fecal contamination and describes the sources, sinks, transport processes, and fate of E. coli contaminants in catchments and associated streams. Spatial features include grazing location, land topography, distance to a nearby stream, and distance through the stream network to the outlet. Temporal features are population dynamics on the land surface, in flow, and on streambeds. The model applies the principles of conservation of mass balance on two different types of pools: grid cells on land surfaces and networked stream segments. The model aims to improve the prediction of the effects of different land management strategies on the fecal contamination of waterways. This is achieved by characterizing the movement of fecal contaminants from land to streams and in-stream mobilization. Processes of attenuation, diffusion, and transport govern the movement. Our study site is a hill land catchment with an area of 140 ha and is used exclusively for animal grazing. The model was calibrated with previous research results, and then tested using the data collected at the outlet of the catchment. The sensitivity of the model predictions was analyzed for different scenarios: effect of stock rate, attenuation rate, and flow volumes. The similar pattern between monitored and predicted E. coli concentration proved that the model captures the key features that control the population dynamics of fecal contaminants. Further experiments are required to expand the model's functionality for covering more mitigation options.     

Enumeration of waterborne Escherichia coli with petrifilm plates: comparison to standard methods

Escherichia coli is often monitored in environmental waters as an indicator of the possible presence of human pathogens associated with feces. Petrifilm E. coli/coliform count plates (3M, Minneapolis, MN), previously validated for enumerating E. coli in food, were tested for monitoring E. coli in environmental water. Escherichia coli counts in environmental water samples enumerated with Petrifilm were significantly correlated (R > 0.9; slope = 0.9-1.0; p < 0.001) with counts obtained with three commonly used methods, mTEC (Becton Dickinson, Sparks, MD), m-ColiBlue (Hach, Loveland, CO), and Colilert-18/IDEXX Quanti-Tray 2000 (IDEXX, Westbrook, ME). Blue colonies on Petrifilm plates were most reliably identified as E. coli when accompanied by gas formation, as determined by characterization of the colonies on MacConkey agar plates (PML Microbiologicals, Mississauga, ON, Canada) and by polymerase chair reaction (PCR) with E. coli-specific primers. The main disadvantage of Petrifilm plates for environmental water testing is the small volume (1 mL per sample) that can be tested; however, the plates appear to be suitable for screening and locating sites that exceed criteria for total body and partial body contact. Simplicity of use and storage, reliability, and relatively low cost make Petrifilm plates suitable for volunteer-based and educational water quality monitoring applications, particularly when used as a preliminary screening method to identify problem sites.     

Transport of Escherichia coli in sand columns with constant and changing water contents

Understanding how changes in volumetric water content (theta) affect bacterial adsorption could help reduce transport of pathogenic and indicator bacteria that may be present in infiltrating wastewater. Three flow regimes that simulated infiltration from a household septic system were evaluated: saturated, unsaturated with a constant volumetric water content theta (constant unsaturated flow), and unsaturated with cyclic changes in theta (variable unsaturated flow). Escherichia coli was suspended in artificial sewage (AS) and applied as step inputs to sand columns, with regular interruptions in input for variable unsaturated flow. A transport model was fit to the saturated and constant unsaturated flow breakthrough curves to determine retardation (R), the first-order filtration coefficient (mu), and the maximum outflow relative concentration (Cmax). The total cells transported as a fraction of input (tau) in all three flow regimes was calculated. Constant unsaturated flow resulted in a significantly lower Cmax (0.633) in comparison with saturated flow (0.803, P < or = 0.05), although unsaturated mu (0.0693 h(-1)) was not significantly different from saturated mu (0.0259 h(-1)). Constant unsaturated flow also resulted in a significantly smaller tau (0.617) than saturated (0.806) or variable unsaturated flow (0.734). In variable unsaturated flow, cell concentrations were out of phase with theta--as the column drained, cell concentrations in the outflow increased; and when a pulse of suspension was applied, cell concentrations decreased. Constant unsaturated flow is probably the best for removal of pathogenic bacteria because this regime resulted in lower maximum concentrations of E. coli and greater cell removal, in comparison with saturated and variable unsaturated flow.     

Efficacy of Bacteroides measurements for reducing the statistical uncertainty associated with hydrologic flow and fecal loads in a mixed use watershed

This paper presents an analysis of the occurrence and uncertainty of source-specific Bacteroides and Escherichia coli in a stream in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region during baseflow conditions. The objectives of the study were to evaluate the occurrence, hydrologic significance, and source of fecal mass in the stream using assays for total Bacteroides (AllBac) and bovine-specific Bacteroides (BoBac), and then to compare these measurements with E. coli densities and loads. Samples were collected during baseflow conditions over several months at seven different main channel sites in the Stock Creek watershed, a 49.3 km2 basin located in Knoxville, TN (USA). We determined instantaneous loads for total fecal loads, bovine fecal loads, and E. coli from measured flow rates and the representative Bacteroides fecal masses and/or E. coli densities. The study indicated a strong correlation between total fecal load (kg d(-1)), bovine fecal load (kg d(-1)), E. coli load rate (CFU d(-1)), 7-d antecedent precipitation, and turbidity. The various datasets were used to establish parameter correlations and spatial dependencies throughout the watershed. The data analysis demonstrated two prevalent patterns throughout the watershed: (i) a runoff-dominated transport and occurrence; and (ii) potential groundwater-dominated transport and occurrence.     

Identifying fecal sources in a selected catchment reach using multiple source-tracking tools

Given known limitations of current microbial source-tracking (MST) tools, emphasis on small, simple study areas may enhance interpretations of fecal contamination sources in streams. In this study, three MST tools-Escherichia coli repetitive element polymerase chain reaction (rep-PCR), coliphage typing, and Bacteroidales 16S rDNA host-associated markers-were evaluated in a selected reach of Plum Creek in south-central Nebraska. Water-quality samples were collected from six sites. One reach was selected for MST evaluation based on observed patterns of E. coli contamination. Despite high E. coli concentrations, coliphages were detected only once among water samples, precluding their use as a MST tool in this setting. Rep-PCR classification of E. coli isolates from both water and sediment samples supported the hypothesis that cattle and wildlife were dominant sources of fecal contamination, with minor contributions by horses and humans. Conversely, neither ruminant nor human sources were detected by Bacteroidales markers in most water samples. In bed sediment, ruminant- and human-associated Bacteroidales markers were detected throughout the interval from 0 to 0.3 m, with detections independent of E. coli concentrations in the sediment. Although results by E. coli-based and Bacteroidales-based MST methods led to similar interpretations, detection of Bacteroidales markers in sediment more commonly than in water indicates that different tools to track fecal contamination (in this case, tools based on Bacteroidales DNA and E. coli isolates) may have varying relevance to the more specific goal of tracking the sources of E. coli in watersheds. This is the first report of simultaneous, toolbox approach application of a library-based and marker-based MST analyses to flowing surface water.     

Risk-based evaluation of Escherichia coli monitoring data from undisinfected drinking water

Drinking water regulations in the United States and elsewhere are based on the occurrence of fecal indicator bacteria. Though not meeting all the criteria of an ideal indicator, nonpathogenic strains of Escherichia coli (E. coli) are used worldwide as an indicator of potential fecal contamination for drinking water and for distribution systems. This is, in part, because real illnesses are related to human pathogens, such as E. coli O157:H7, whose presence may be predicted better by E. coli than by total coliform bacteria. Our objective was to estimate the number of E. coli O157:H7 illnesses attributable to drinking water exposures in the United States and the feasible relationships between positive occurrences of the indicator bacteria E. coli and E. coli O157:H7 in drinking water. Results of the modeling indicate that in undisinfected drinking water systems, the ratio of bacterial indicator E. coli positives to E. coli O157:H7 organisms is estimated to be between 6:1 and 90:1 with few model parameters accounting for the vast majority of the uncertainty. These results provide context for considering the potential public health implications of a positive E. coli result from routine monitoring of undisinfected drinking water.     

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.     

Effect of manure on Escherichia coli attachment to soil

Attachment of bacteria to soil is an important component of bacterial fate and transport. Escherichia coli are commonly used as indicators of fecal contamination in the environment. Despite the fact that E. coli are derived exclusively from feces or manure, effect of the presence of manure colloids on bacteria attachment to agricultural soils was never directly studied. The objective of this work was to evaluate the magnitude of the effect of manure on E. coli attachment to soil. Escherichia coli attachment to soil was studied in batch experiments with samples of loam and sandy clay loam topsoil that were taken in Pennsylvania and Maryland. Escherichia coli cells were added to the water-manure suspensions containing 0, 20, and 40 g L(-1) of filtered liquid bovine manure, which subsequently were equilibrated with air-dry sieved soil in different soil to suspension ratios. The Langmuir isotherm equation was fitted to data. Manure dramatically affected E. coli attachment to soil. Attachment isotherms were closer to linear without manure and were strongly nonlinear in the presence of manure. The maximum E. coli attachment occurred in the absence of manure. Increasing manure content generally resulted in decreased attachment.     

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。     

Spatial and temporal drivers of zoonotic pathogen contamination of an agricultural watershed

In regions where animal agriculture is prominent, such as southern Alberta, higher rates of gastrointestinal illness have been reported when compared with nonagricultural regions. This difference in the rate of illness is thought to be a result of increased zoonotic pathogen exposure through environmental sources such as water. In this study, temporal and spatial factors associated with bacterial pathogen contamination of the Oldman River, which transverses this region, were analyzed using classification and regression tree analysis. Significantly higher levels of fecal indicators; more frequent isolations of Campylobacter spp., Escherichia coli O157:H7, and Salmonella enterica spp.; and higher rates of detection of pig-specific Bacteroides markers occurred at downstream sites than at upstream sites, suggesting additive stream inputs. Fecal indicator densities were also significantly higher when any one of these three bacterial pathogens was present and where there were higher total animal manure units; however, occasionally pathogens were present when fecal indicator levels were low or undetectable. Overall, Salmonella spp., Campylobacter spp., and E. coli O157:H7 presence was associated with season, animal manure units, and total rainfall on the day of sampling and 3 d in advance of sampling. Several of the environmental variables analyzed in this study appear to influence pathogen prevalence and therefore may be useful in predicting water quality and safety and in the improvement of watershed management practices in this and other agricultural regions.     

FECAL‐INDICATOR BACTERIA IN STREAMS ALONG A GRADIENT OF RESIDENTIAL DEVELOPMENT1

ABSTRACT: Fecal‐indicator bacteria were sampled at 14 stream sites in Anchorage, Alaska, USA, as part of a study to determine the effects of urbanization on water quality. Population density in the subbasins sampled ranged from zero to 1,750 persons per square kilometer. Higher concentrations of fecal‐coliform, E. coli, and enterococci bacteria were measured at the most urbanized sites. Although fecal‐indicator bacteria concentrations were higher in summer than in winter, seasonal differences in bacteria concentrations generally were not significant. Areas served by sewer systems had significantly higher fecal‐indicator bacteria concentrations than did areas served by septic systems. The areas served by sewer systems also had storm drains that discharged directly to the streams, whereas storm sewers were not present in the areas served by septic systems. Fecal‐indicator bacteria concentrations were highly variable over a two‐day period of stable streamflow, which may have implications for testing of compliance to water‐quality standards.     

Solar and Tidal Modulations of Fecal Indicator Bacteria in Coastal Waters at Huntington Beach,California

The coastal waters at many beaches in California and the United States are afflicted with fecal pollution, which poses a health risk for people exposed to the water through recreational activities such as swimming, surfing, and diving. Identifying sources of pollution is complicated by oceanographic transport/mixing processes and the nonconservative behavior of microorganisms exposed to sunlight and hostile marine conditions. This article investigates the variation of fecal indicator bacteria (FIB) concentrations in the surf zone and the adjacent coastal marsh by applying autocorrelation and cross-correlation analyses that illustrate solar and tidal modulations. A steady state bioreactor model was developed to explain solar inactivation in the surf zone, whereas a dynamic model was applied to explain tidally influenced disturbances in the coastal marsh. These models applied to intensive monitoring datasets on FIB and environmental variables have provided insights into the biologic and physical processes controlling coastal water quality, specifically the influence of sunlight and tides on bacterial levels.     

Investigation of an Escherichia coli environmental benchmark for waterborne pathogens in agricultural watersheds in Canada

Canada's National Agri-Environmental Standards Initiative sought to develop an environmental benchmark for low-level waterborne pathogen occurrence in agricultural watersheds. A field study collected 902 water samples from 27 sites in four intensive agricultural watersheds across Canada from 2005 to 2007. Four of the sites were selected as reference sites away from livestock and human fecal pollution sources in each watershed. Water samples were analyzed for Campylobacter spp., Salmonella spp., Escherichia coli O157:H7, Cryptosporidium spp., Giardia spp., and the water quality indicator E. coli. The annual mean number of pathogen species was higher at agricultural sites (1.54 ± 0.07 species per water sample) than at reference sites (0.75 ± 0.14 species per water sample). The annual mean concentration of E. coli was also higher at agricultural sites (491 ± 96 colony-forming units [cfu] 100 mL(-1)) than at reference sites (53 ± 18 cfu 100 mL(-1)). The feasibility of adopting existing E. coli water quality guideline values as an environmental benchmark was assessed, but waterborne pathogens were detected at agricultural sites in 80% of water samples with low E. coli concentrations (<100 cfu 100 mL(-1)). Instead, an approach was developed based on using the natural background occurrence of pathogens at reference sites in agricultural watersheds to derive provisional environmental benchmarks for pathogens at agricultural sites. The environmental benchmarks that were derived were found to represent E. coli values lower than geometric mean values typically found in recreational water quality guidelines. Additional research is needed to investigate environmental benchmarks for waterborne pathogens within the context of the "One World, One Health" perspective for protecting human, domestic animal, and wildlife health.     

Putative temporal variability of Escherichia coli ribotypes from yearling steers

Escherichia coli is a ubiquitous component of the intestinal microflora of warm-blooded animals, and is an indicator of fecal contamination of surface waters. Ribotype profiling of E. coli is one of several genotypic methods that has been developed to determine the host origin of fecal bacteria. Like most genotypic methods of source tracking, ribotyping requires a host origin database to identify environmental isolates. To determine the extent of temporal variability of ribotypes and its effect on a host origin database, E. coli isolates were obtained from fecal samples of two herds of Black Angus steers at a long-term experimental site at four sampling times from October 1999 to July 2000. Fecal samples were taken from six randomly chosen steers at each time. At a similarity index of 90% as calculated by unweighted pair-group method using arithmetic averages (UPGMA), 240 ribotypes were identified from 451 E. coli isolates. Only 20 ribotypes (8.3%), comprising 33% of the total isolates, were shared among sampling times and were considered resident ribotypes. Two of the twenty resident ribotypes appeared at three sampling times, and the remaining eighteen appeared at two. The majority of the ribotypes, therefore, were transient and unique to each sampling time and steer. Both the apparent turnover of E. coli ribotypes and a clonal diversity index of 0.97 (indicative of extensive ribotype variability) suggest the necessity of ribotyping a large number E. coli isolates per host to establish a host origin database that is independent of temporal variability, or complete enough to be effective.     

Oxidative destabilization of dissolved organics and E. coli in domestic wastewater through immobilized cell reactor system

Domestic wastewater contains a considerable amount of pathogenic organisms besides non-biodegradable organics. The conventional technologies followed for the treatment of domestic wastewater are less efficient in removing pathogenic organisms despite substantial removal of dissolved organics. The focal theme of the present investigation was to use a chemo-autotrophic activated carbon oxidation (CAACO) system, an immobilized cell reactor using chemoautotrophs (Bacillus sp.) for the treatment of domestic wastewater. The oxidation of organics and Escherichia coli in wastewater is controlled by the parameters space time, O(2)/COD, bed height and cod loading. The scheme comprised of anaerobic treatment, sand filtration and CAACO treatment removed BOD. COD, Total organic carbon (TOC), dissolved protein, total Kjeldhal nitrogen (TKN) and bacterial count (most probable number (MPN)) by 81%, 92%, 84%, 94%, 93% and 99.9997%, respectively. The low concentration of E. coli in the CAACO-treated wastewater was completely eliminated through UV irradiation in 3 min at 254 nm.     

Wildlife identified as major source of Escherichia coli in agriculturally dominated watersheds by BOX A1R-derived genetic fingerprints

The presence of Escherichia coli in recreational and potable waters is a major concern to the general public as elevated levels of E. coli suggest the presence of pathogenic bacteria and viruses. Unfortunately, traditional microbial techniques do not allow specific identification of the source of E. coli. This reduces the ability to target management practices that reduce bacterial contamination. In the Finger Lakes region of western New York, USA, wildlife resides in relatively high densities on watersheds dominated by people and dairy farms, and as a result, the sources of fecal degradation of potable and recreational waters are often unknown. In the Conesus Lake watershed, the sources of microbial contamination were assessed using Rep-PCR molecular tools, a method of amplifying repetitive DNA sequences found throughout the E. coli genome to produce distinct fingerprints for a given ecotype. Molecular fingerprints of E. coli isolated from regional populations of cattle, humans, geese and deer were compared to E. coli isolated from stream water samples. Canonical discriminant function analysis indicated that the DNA fingerprints of the original source group isolates were correctly predicted 90.2% of the time. Since land use in the sub-watersheds was dominated by dairy and cash crop farms, it was expected that the majority of E. coli isolated would be identified as cows; however, an unexpectedly high percentage of isolates were identified as wildlife (geese and deer). Geese were the dominant source of E. coli (44.7-73.7% of the total sources) in four sub-watersheds followed by cows (10.5-21.1%), deer (10.5-18.4%), humans (5.3-12.9%) and unidentifiable sources (0.0-11.8%). Management practices intended to decrease the number of cattle or the amount of manure spread in a sub-watershed were reflected in a decrease of E. coli ecotypes associated with dairy cows.     

Biochar pyrolyzed at two temperatures affects Escherichia coli transport through a sandy soil

The incorporation of biochar into soils has been proposed as a means to sequester carbon from the atmosphere. An added environmental benefit is that biochar has also been shown to increase soil retention of nutrients, heavy metals, and pesticides. The goal of this study was to evaluate whether biochar amendments affect the transport of Escherichia coli through a water-saturated soil. We looked at the transport of three E. coli isolates through 10-cm columns packed with a fine sandy soil amended with 2 or 10% (w/w) poultry litter biochar pyrolyzed at 350 or 700°C. For all three isolates, mixing the high-temperature biochar at a rate of 2% into the soil had no impact on transport behavior. When added at a rate of 10%, a reduction of five orders of magnitude in the amount of E. coli transported through the soil was observed for two of the isolates, and a 60% reduction was observed for the third isolate. Mixing the low-temperature biochar into the soil resulted in enhanced transport through the soil for two of the isolates, whereas no significant differences in transport behavior were observed between the low-temperature and high-temperature biochar amendments for one isolate. Our results show that the addition of biochar can affect the retention and transport behavior of E. coli and that biochar application rate, biochar pyrolysis temperature, and bacterial surface characteristics were important factors determining the transport of E. coli through our test soil.