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.     

Phosphorus, sediment, and Escherichia coli loads in unfenced streams of the Georgia Piedmont, USA

Contamination of unfenced streams with P, sediments, and pathogenic bacteria from cattle (Bos taurus) activity may be affected by the availability of shade and alternative water sources. The objectives of this study were to evaluate water quality in two streams draining tall fescue (Festuca arundinacea Schreb.)-common bermudagrass (Cynodon dactylon L.) pastures with different shade distribution, and to quantify the effects of alternative water sources on stream water quality. For 3 yr, loads of dissolved reactive phosphorus (DRP), total phosphorus (TP), and total suspended solids (TSS) were measured during storm flow, and loads of DRP, TP, TSS, and Escherichia coli were measured every 14 d during base flow. We also used GPS collars to determine amount of time cattle spent in riparian areas. Our results showed that cattle-grazed pastures with unfenced streams contributed significant loads of DRP, TP, TSS, and E. coli to surface waters (p < 0.01). Time spent by cattle in riparian areas as well as storm flow loads of DRP, TP, and TSS were larger (p < 0.08) in the pasture with the smaller amount of nonriparian shade. Water trough availability decreased base flow loads of TSS and E. coli in both streams, and decreased time cattle spent in riparian areas in the pasture with the smaller amount of nonriparian shade (p < 0.08). Our results indicate that possible BMPs to reduce contamination from cattle-grazed pastures would be to develop or encourage nonriparian shade and to provide cattle with alternative water sources away from the stream.     

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.     

Transfer of Escherichia coli to water from drained and undrained grassland after grazing

The aim of this study was to determine the load of Escherichia coli transferred via drainage waters from drained and undrained pasture following a grazing period. Higher concentrations (ranging between 10(4) and 10(3) colony forming units [CFU] g(-1)) of E. coli persisted in soil for up to 60 d beyond the point where cattle were removed from the plots, but these eventually declined in the early months of spring to concentrations less than 10(2) CFU g(-1). The decline reflects the combined effect of cell depletion from the soil store through both wash-out and die-off of E. coli. No difference (P > 0.05) was observed in E. coli loads exported from drained and undrained plots. Similarly, no difference (P > 0.05) was observed in E. coli concentrations in drainage waters of mole drain flow and overland plus subsurface interflow. Intermittent periods of elevated discharge associated with storm events mobilized E. coli at higher concentrations (e.g., in excess of 400 CFU mL(-1)) than observed during low flow conditions (often <25 CFU mL(-1)). The combination of high discharge and cell concentrations resulted in the export of E. coli loads from drained and undrained plots exceeding 10(6) CFU L(-1) s(-1). The results highlight the potential for drained land to export E. coli loads comparable with those transferred from undrained pasture.     

Resuspension of sediment-associated Escherichia coli in a natural stream

In this study, a tracer bacteria was used to investigate the resuspension and persistence of sediment-associated bacteria in a small alluvial stream. The study was conducted in Swan Creek, located within the Grand River watershed of Ontario, Canada. A 1.1-m2 section of the bed was seeded with a strain of Escherichia coli resistant to nalidixic acid (E. coli NAR). The survival, transport, and redistribution of the tracer bacteria within a 1.7-km river section downstream of the source cell was assessed for a 2-mo period following the introduction of the tracer bacteria. This study has illustrated that enteric bacteria can survive in bed sediments for up 6 wk and that inactivation of the tracer bacteria resembled typical first-order decay. Critical conditions for resuspension, as well as resuspension rates, of sediment-associated bacteria were determined for several storm events. The critical shear stress for E. coli NAR resuspension in Swan Creek ranged from 1.5 to 1.7 N m(-2), which is comparable with literature values for critical shear stresses for erosion of cohesive sediments. Bacteria resuspension was primarily limited to the rising limb of storm hydrographs implying that a finite supply of sediment-associated bacteria are available for resuspension during individual storm events. The information presented in this paper will further the development of representative microbial water quality models.     

Population Dynamics of Escherichia coli in Surface Water1

Bucci, Vanni, Marin Vuli?, Xiaodan Ruan, and Ferdi L. Hellweger, 2011. Population Dynamics of Escherichia coli in Surface Water. Journal of the American Water Resources Association (JAWRA) 47(3):611‐619. DOI: 10.1111/j.1752‐1688.2011.00528.x Abstract: Present models of the fate of enteric bacteria in surface water usually assume simple first‐order decay. However, observed decay kinetics are typically biphasic and there is increasing evidence for growth in the ambient environment. These features are similar to the stationary phase of long‐term batch cultures, where the dynamics are the result of repeated cycles of population takeovers by Growth Advantage in Stationary Phase (GASP) mutants able to scavenge nutrients released from dead cells. We investigated the possible role of this mechanism (i.e., growth of an adapted subpopulation) in surface water using laboratory experiments with various strains of Escherichia coli in different types of water. Addition of ampicillin (which only kills dividing cells) to these cultures caused a decrease in the total cell density, demonstrating the presence of a growing subpopulation. Furthermore, long‐term survivors from surface water cultures outcompete naïve cells and take over the population in surface water. This growth advantage is heritable (i.e., aged strains were isolated from a single cell) and consistent with the GASP mechanism, although the responsible loci were not identified. Our results show that E. coli populations in surface water are dynamic and consist of dying naïve cells and growing surface water adapted cells. These results can explain the apparent biphasic decay pattern observed in laboratory experiments. This mechanism may also be important in the ambient environment.     

Comparison of Escherichia coli and Campylobacter jejuni transport in saturated porous media

Due to the difficulties in testing for specific pathogens, water samples are tested for the presence of nonpathogenic indicator organisms to determine whether a water supply has been contaminated by fecal material. An implicit assumption in this approach is that where pathogenic microorganisms are present fecal indicator organisms are present as well; yet surprisingly few studies have been conducted that directly compare the transport of indicator organisms with pathogenic organisms in ground water environments. In this study we compared the cell properties and transport of Escherichia coli, a commonly used indicator organism, and Campylobacter jejuni, an important enteropathogen commonly found in agricultural wastes, through saturated porous media. Differences in cell properties were determined by measuring cell geometry, hydrophobicity, and electrophoretic mobility. Transport differences were determined by conducting miscible displacement experiments in laboratory columns. Under the experimental conditions tested, C. jejuni was much more negatively charged and more hydrophobic than E. coli. In addition, C. jejuni cells were slightly longer, narrower, and less spherical than E. coli. The variations in cell properties, primarily surface charge, resulted in significant differences in transport between these two microorganisms, with the transport of C. jejuni exceeding that of E. coli when conditions favored low attachment rates, thus calling into question the usefulness of using E. coli as an indicator organism for this important pathogen.     

Escherichia coli load reduction from runoff by vegetative filter strips: a laboratory-scale study

Vegetative filter strips (VFS) are commonly used best management practices for removing contaminants from runoff. Additional research is warranted to determine their efficiency and the most appropriate metrics for predicting fecal bacteria reductions. The objective of this research was to determine VFS effectiveness in removing from runoff relative to inflow rate, infiltration capacity, and flow concentration. This research also investigated the presence of in runoff from clean water runon after diluted manure runon events. A laboratory-scale VFS soil box (200 cm long, 100 cm wide, 7.5% slope) was packed with a sandy loam soil. Ten constant-flow VFS experiments were conducted with and without vegetation (8-10 cm ryegrass [ L.]) at low (20-40 cm s), medium (40-60 cm s), and high (85-120 cm s) flow rates and for a full (100 cm) or concentrated (40 cm) VFS flow width to simulate a channelizing flow condition. Two runon events were investigated for each experimental condition: (i) diluted liquid swine manure runon and (ii) clean water runon 48 h afterward. was used as an indicator of fecal contamination and was quantified by the most probable number (MPN) technique. No concentration reductions were observed based on peak outflow concentrations, and only small concentration reductions were observed based on outflow event mean concentrations. The mass reductions ranged from 22 to 71% and were strongly correlated to infiltration or runoff reduction ( = 0.88), which was dependent on the degree of flow concentration. Little to no effect of sedimentation on transport was observed, hypothesized to be due to minimum attachment to sediment particles because the bacteria originated from manure sources. Therefore, the design of VFS for bacteria removal should be based on the infiltration capacity in the VFS and should prevent concentrated flow, which limits total infiltration. The event mean concentrations in clean water runon experiments were between 10 and 100 MPN per 100 mL; therefore, under these conditions, VFS served as a source of residual from previous runon events.     

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.     

Locating Escherichia coli contamination in a rural South Carolina watershed

One of the problems associated with the use of ambient water quality standards in surface water regulation is the difficulty of identifying and regulating nonpoint source pollution, making such standards unenforceable, especially at the local level. We used the Escherichia coli indicator to locate the most contaminated reaches in rural South Carolina's Bush River watershed (297 km(2), 186 stream-km). We divided the watershed into 20 smaller reaches and sampled each reach multiple times, but restricted each sampling round to one day. We located four low order creek reaches, representing just nine stream-km, where we observed geometric mean E. coli densities of over 1250 E. coli/100 mL; in each case, the source of the contamination (riparian grazing of cattle) was easily identifiable. On the Bush River itself, we observed a step change in one reach where geometric means increased from 106 E. coli/100 mL to 565 E. coli/100 mL over the reach's 10 km length. In this case, the sources of contamination were not as obvious as in the lower order streams; in this case, more advanced Microbial Source Tracking techniques will be required to identify the sources. Nevertheless, this sampling protocol helped locate polluted reaches and provided decision-makers with reasonable justifications for concrete action in deciding where (or where not) to install conservation practices and where more sophisticated (and expensive) MST techniques were warranted.     

Using zebra mussels to monitor Escherichia coli in environmental waters

Use of the zebra mussel (Dreissena polymorpha) as an indicator of previously elevated bacteria concentrations in a watershed was examined. The ability of the zebra mussel to accumulate and purge Escherichia coli over several days was investigated in both laboratory and field experiments. In laboratory experiments, periodic enumeration of E. coli in mussels that had been exposed to a dilute solution of raw sewage demonstrated that (i) maximum concentrations of E. coli are reached within a few hours of exposure to sewage, (ii) the tissue concentration attained is higher than the concentration in the ambient water, and (iii) the E. coli concentrations take several days to return to preexposure concentrations when mussels are subsequently placed in sterile water. In field experiments conducted in southeast Michigan in the Clinton River watershed, brief increases in E. coli concentrations in the water were accompanied by increases in mussel concentrations of E. coli that lasted 2 or 3 d. The ability of mussels to retain and to concentrate E. coli made it possible to detect E. coli in the environment under conditions that conventional monitoring may often miss. Sampling caged mussels in a river and its tributaries may enable watershed managers to reduce the sampling frequency normally required to identify critical E. coli sources, thereby providing a more cost-effective river monitoring strategy for bacterial contamination.     

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.     

Persistence of culturable Escherichia coli fecal contaminants in dairy alpine grassland soils

Our knowledge of Escherichia coli (E. coli) ecology in the field is very limited in the case of dairy alpine grassland soils. Here, our objective was to monitor field survival of E. coli in cow pats and underlying soils in four different alpine pasture units, and to determine whether the soil could constitute an environmental reservoir. E. coli was enumerated by MPN using a selective medium. E. coli survived well in cow pats (10(7) to 10(8) cells g(-1) dry pat), but cow pats disappeared within about 2 mo. In each pasture unit, constant levels of E. coli (10(3) to 10(4) cells g(-1) dry soil) were recovered from all topsoil (0-5 cm) samples regardless of the sampling date, that is, under the snow cover, immediately after snow melting, or during the pasture season (during and after the decomposition of pats). In deeper soil layers below the root zone (5-25 cm), E. coli persistence varied according to soil type, with higher numbers recovered in poorly-drained soils (10(3) to 10(4) cells g(-1) dry soil) than in well-drained soils (< 10(2) cells g(-1) dry soil). A preliminary analysis of 38 partial uidA sequences of E. coli from pat and soils highlighted a cluster containing sequences only found in this work. Overall, this study raises the possibility that fecal E. coli could have formed a naturalized (sub)population, which is now part of the indigenous soil community of alpine pasture grasslands, the soil thus representing an environmental reservoir of E. coli.     

Farm-level irrigation and the marginal cost of water use: evidence from Georgia

We create a proxy for the cost of irrigation water in Georgia from a sample of Georgia irrigators by investigating the marginal cost of pumping groundwater. We then combine this proxy with agronomic and climatic variables to estimate the response of agricultural water use to differences in the marginal cost of irrigation. The results show that pumping costs are a significant determinant of water use, and imply that agricultural water use would be moderately affected by institutional changes that would explicitly price water.     

Escherichia coli loading at or near base flow in a mixed-use watershed

This study analyzed the occurrence of Escherichia coli in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region using synoptic monitoring (samples taken throughout the watershed system) during base-flow conditions. The objective of the study was to evaluate the occurrence of E. coli during base-flow conditions for several months at seven different main channel and nine different tributary sampling sites in the Stock Creek watershed, a 49.3-km(2) basin located in Knoxville, TN. Escherichia coli densities were measured using the Colilert (Defined Substrate Technology) method. The instantaneous loads for E. coli were determined from measured flow rates and E. coli densities, with the highest loading rates observed in the late fall. The study indicated a strong correlation between E. coli load rate (colony-forming units [CFU]/d), 7-d antecedent precipitation, and turbidity. Water quality data, however, also exhibited a spatial dependency; for example, the E. coli load rate was better correlated with turbidity in the slower draining basin tailwater sampling sites than in the faster draining upstream headwater sampling sites. In the headwater sites, the E. coli load rate was better correlated with 7-d antecedent precipitation than turbidity.     

Predicting Streambed Sediment and Water Column Escherichia coli Levels at Watershed Scale

A sub‐model for the Soil and Water Assessment Tool (SWAT) is developed to predict Escherichia coli levels in the streambed sediment as well as in the water column. New formulations to estimate the levels of E. coli in streambed sediment and the water column are derived. These equations include calculations of E. coli resuspension from the streambed sediment to the water column, E. coli deposition from the water column to the streambed sediment, E. coli growth in the streambed sediment and the water column, and instream E. coli routing. These formulations were programmed in FORTRAN and integrated into SWAT. The modified SWAT model was applied to Squaw Creek Watershed, Iowa, to predict E. coli levels in the stream. Escherichia coli concentrations in the streambed sediment and the water column were monitored extensively in this watershed, and observations were used to verify the model predictions. The model proposed here can predict E. coli concentrations in streambed sediment as well as in the water column. Approximately 58% of the predictions of E. coli levels in the bed sediment were within 1 order of magnitude from the observed value, and in the water column 83% of the predictions of E. coli levels were within 1 order of magnitude. Results suggest that the proposed model will help predictions of instream bacterial contamination.     

Sequence-based source tracking of Escherichia coli based on genetic diversity of beta-glucuronidase

High levels of fecal bacteria are a concern for recreational waters; however, the source of contamination is often unknown. This study investigated whether direct sequencing of a bacterial gene could be utilized for detecting genetic differences between bacterial strains for microbial source tracking. A 525-nucleotide segment of the gene for beta-glucuronidase (uidA) was sequenced in 941 Escherichia coli isolates from the Clinton River-Lake St. Clair watershed, 182 E. coli isolates from human and animal feces, and 34 E. coli isolates from a combined sewer. Environmental isolates exhibited 114 alleles in 11 groups on a genetic tree. Frequency of strains from different genetic groups differed significantly (p < 0.03) between upstream reaches (Bear Creek-Red Run), downstream reaches, and Lake St. Clair beaches. Fecal E. coli uidA sequences exhibited 81 alleles that overlapped with the environmental set. An algorithm to assign alleles to different host sources averaged approximately 75% correct classification with the fecal data set. Using the same algorithm, the percent of environmental isolates assignable to humans decreased significantly between Bear Creek-Red Run (30 +/- 3%) and the beaches (17 +/- 2%) (p < 0.05). Birds accounted for approximately 50% of assignable environmental isolates. For combined sewer isolates, the same algorithm assigned 51% to humans. These experiments demonstrate differences in the frequency of different E. coli strains at different locations in a watershed, and provide a "proof in principle" that sequence-based data can be used for microbial source tracking.     

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.     

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.     

Combination of Biological and Habitat Indices for Assessment of Idaho Streams

States and tribes are encouraged to use multiple biological assemblages in assessment of water bodies. An assessment index for each assemblage provides information on aspects of the aquatic resource that may be unique in terms of stressor sensitivity, stressor type, or ecological scale. However, assessment results relative to impairment thresholds can disagree among indices for an individual water body, leading to uncertain overall water‐body assessments. We explored options for combining stream indices for macroinvertebrates, fish, and habitat in ways that would yield the most consistent and sensitive results relative to established disturbance categories. Methods varied in the scoring or rating scales used to standardize each index value, the thresholds used to define impairment of aquatic life uses, and the ways of synthesizing multiple indices. The index compositing method that scores each index on a continuous scale and averages the scores after standardizing had superior accuracy, sensitivity, and precision. In addition, using the 25th quantile of reference sites instead of the 10th quantile resulted in a more balanced error rate among reference and degraded site categories.