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.     

Direct and indirect hydrological controls on E. coli concentration and loading in midwestern streams

This study investigates hydrological controls on E. coli concentration and loading in two artificially drained agricultural watersheds (58 and 23 km(2)) of the U.S. Midwest. Stream E. coli concentrations are significantly (p < 0.02) lower at base flow than high flow; however, E. coli load is significantly higher at high flow than at low flow (p < 0.001). Although E. coli concentrations are not significantly higher (p = 0.253) in summer/fall (3269 MPN/100 mL) than in the winter/spring (2411 MPN/100 mL), E. coli load is significantly higher (p < 0.05) in winter/spring (346 MPN/day) than in summer/fall season (75 MPN/day). Correlation analysis indicates that discharge and precipitation are the best indicators of E. coli concentration and 7-d antecedent precipitation (7dP), the best indicator of E. coli loading in the watersheds studied regardless of flow conditions and location. However, E. coli concentration and loading best correlate to 7dP and turbidity at base flow. A spatial dependency is also observed at base flow with E. coli concentration and load correlating better to 7dP in the headwaters and to turbidity in the lower reaches of the watersheds studied. For high flow conditions, E. coli concentration and loading are poorly correlated to most variables, except stream water temperature and 7-d antecedent discharge. These results are consistent with those reported in the literature and suggest that, at least during base flow conditions, turbidity and 7dP may be usable in artificially drained landscapes of the Midwest to identify potential hot spots of E. coli contamination.     

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.     

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.     

Demonstration of methods to reduce E. coli runoff from dairy manure application sites

Contamination by bacteria is a leading cause of impairment in U.S. waters, particularly in areas of livestock agriculture. We evaluated the effectiveness of several practices in reducing Escherichia coli levels in runoff from fields receiving liquid dairy (Bos taurus) manure. Runoff trials were conducted on replicated hay and silage corn (Zea mays L.) plots using simulated rainfall. Levels of E. coli in runoff were approximately 10(4) to 10(6) organisms per 100 mL, representing a significant pollution potential. Practices tested were: manure storage, delay between manure application and rainfall, manure incorporation by tillage, and increased hayland vegetation height. Storage of manure for 30 d or more consistently and dramatically lowered E. coli counts in our experiments, with longer storage providing greater reductions. Manure E. coli declined by > 99% after approximately 90 d of storage. On average, levels of E. coli in runoff were 97% lower from plots receiving 30-d-old and > 99% lower from plots receiving 90-d-old manure than from plots where fresh manure was applied. Runoff from hayland and cornland plots where manure was applied 3 d before rainfall contained approximately 50% fewer E. coli than did runoff from plots that received manure 1 d before rainfall. Hayland vegetation height alone did not significantly affect E. coli levels in runoff, but interactions with rainfall delay and manure age were observed. Manure incorporation alone did not significantly affect E. coli levels in cornland plot runoff, but incorporation could reduce bacteria export by reducing field runoff and interaction with rainfall delay was observed. Extended storage that avoids additions of fresh manure, combined with application several days before runoff, incorporation on tilled land, and higher vegetation on hayland at application could substantially reduce microorganism loading from agricultural land.     

Significant Escherichia coli attenuation by vegetative buffers on annual grasslands

A study was conducted to estimate the retention efficiency of vegetative buffers for Escherichia coli deposited on grasslands in cattle fecal deposits and subject to natural rainfall-runoff conditions. The study was conducted on annual grasslands in California's northern Sierra Nevada foothills, a region with a distinct wet-dry season Mediterranean climate. We used 48, 2.0- by 3.0-m runoff plots to examine the efficacy of 0.1-, 1.1-, and 2.1-m buffers at three land slopes (5, 20, and 35%) and four dry vegetation matter levels (225, 560, 900, and 4500 kg/ha) across 27 rainfall-runoff events during two rainfall seasons. Buffer width treatments were implemented by placement of cattle fecal material containing known loads of E. coli 0.1, 1.1, or 2.1 m upslope of the plot runoff collector. Mean total runoff to total rainfall ratio per plot ranged from 0.014:1 to 0.019:1 and reflected the high infiltration capacity of these soils. Approximately 94.8 to 99.995% of total E. coli load applied to each plot appears to be either retained in the fecal pat and/or attenuated within 0.1 m downslope of the fecal pat, irrespective of the presence of a wider vegetated buffer. Relative to a 0.1-m buffer, we found 0.3 to 3.1 log10 reduction in E. coli discharge per additional meter of vegetative buffer across the range of residual dry vegetation matter levels, land slope, and rainfall and runoff conditions experienced during this project. Buffer efficiency was significantly reduced as runoff increased. These results support the assertion that grassland buffers are an effective method for reducing animal agricultural inputs of waterborne E. coli into surface waters.     

Bedding and seasonal effects on chemical and bacterial properties of feedlot cattle manure

Nutrients, soluble salts, and pathogenic bacteria in feedlot-pen manure have the potential to cause pollution of the environment. A three-year study (1998-2000) was conducted at a beef cattle (Bos taurus) feedlot in southern Alberta, Canada to determine the effect of bedding material [barley (Hordeum vulgare L.) straw versus wood chips] and season on the chemical and bacterial properties of pen-floor manure. Manure was sampled for chemical content (N, P, soluble salts, electrical conductivity, and pH) and populations of four groups of bacteria (Escherichia coli, total coliforms, and total aerobic heterotrophs at 27 and 39 degrees C). More chemical parameters of manure were significantly (P < or = 0.05) affected by season (SO4, Na, Mg, K, Ca, sodium adsorption ratio [SAR], total C, NO3-N, NH4-N, total P, and available P) than by bedding (K, pH, total C, C to N ratio, NH4-N, and available P). Bedding had no significant (P > 0.05) effect on the four bacterial groups whereas season affected all four groups. Numbers of E. coli and total coliforms (TC) were significantly higher by 1.72 to 2.02 log10 units in the summer than the other three seasons, which was consistent with a strong positive correlation of E. coli and TC with air temperature. The low ratio of bedding to manure in the pens was probably the major cause of the lack of significant bedding effects. Bedding material and seasonal timing of cleaning feedlot pens and land application of manure may be a potential tool to manage nutrients, soluble salts, and pathogens in manure.     

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.     

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.     

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.     

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.     

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.     

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.     

Riparian Vegetation and Water Chemistry in a Basin Under Semiarid Mediterranean Climate, Andarax River, Spain

A study has been made of the relationships between the characteristics of the riparian vegetation (floristic composition, structure and diversity) and the spatial–temporal variation of the quality of the stream waters in a basin under a semiarid Mediterranean climate in the southeastern Iberian Peninsula. The plant communities of the high reaches present greater specific richness and diversity (S _mean= 7.0 ± 3.4 and H′_mean= 2.0 ± 0.7) than do those of the middle and low reaches (S _mean= 4.5 ± 1.6 and H′_mean= 1.8 ± 0.6). One zone reached the highest specific richness (S= 12, H′= 3.2), which, apart from being situated in the intermediate stretch of the basin, represents a transitional state (ecotone) between the Salix and Tamarix communities. The characteristics of the waters analyzed indicate very high rates of erosion and runoff due to the nature of the soils (easily eroded marls) and to agricultural expansion and mining since the 16th century. The present-day riparian vegetation is not adequate to absorb the nitrates added to the basin by crop fertilization, reaching extremely high values, particularly during the dry period (between 1.2 and 42.5 mg/liter). Sewage dumping at three sampling stations did not appear to affect the specific composition of the woody vegetation. In the zones with watercourses, water salinity was low during the period of greater water flow, but considerably higher in the dry season (the upper limit was some 1.2 mS/m), resulting in a predominance of salt cedars over willows. Three types of saltcedar areas were distinguished: subhalophilous, which barely changes its chemical composition over the season; halophilous, which develops over strongly mineralized waters and markedly alters in chemical composition during the dry season; and hyperhalophilous, where salinity is extraordinarily high and quite constant throughout the year. A direct relationship was found between the dominance of Tamarix africana and abundance of NaCl.     

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.     

Removal of faecal bacteria from septage by treating it in a full-scale duckweed-covered pond system

Performance of a full-scale duckweed-covered treatment system in removing faecal bacteria is presented. The system consisted of three ponds in series and received septage from holding tanks. Inflow averaged between 36 m³ d⁻¹ in the cold season and 60 m³ d⁻¹ in the warm season, resulting in a total hydraulic retention time of 88 and 58 days, respectively. Duckweed (Lemna minor) colonized the ponds in the summer and continued to grow in the cold season. Due to the difficult harvesting process of the duckweed biomass, the investigation of the treatment efficiency was carried out without plant harvesting. The system was monitored for temperature, pH, oxygen, chlorophyll-a, Escherichia coli and Enterococci. Duckweed growth resulted in chlorophyll-a concentration reduction from 924 to 13 μg L⁻¹, causing neutral and anoxic conditions in the pond water. A temperature effect was noticed on the E. coli decay coefficient with a decreasing trend along the treatment system. Enterococci always decayed less than E. coli. Differences on decay coefficients and removal efficiencies were not observed between the three ponds for both bacterial types. Effluent quality in terms of E. coli was 489 and 1377 cfu/100 mL, in the warm and the cold seasons, respectively, with average removals of 99.65 ± 1.46% and 99.33 ± 3.03%. Total Enterococci removal was 88.91 ± 23.1% in the warm season and 94.43 ± 24.45% in the cold season, resulting in mean effluent values of 1058 and 1404 cfu/100 mL, respectively. The seasonal differences in total removal efficiencies were insignificant for both bacterial types.     

Incidence and persistence of zoonotic bacterial and protozoan pathogens in a beef cattle feedlot runoff control vegetative treatment system

Determining the survival of zoonotic pathogens in livestock manure and runoff is critical for understanding the environmental and public health risks associated with these wastes. The occurrence and persistence of the bacterial pathogens Escherichia coli O157:H7 and Campylobacter spp. in a passive beef cattle feedlot runoff control-vegetative treatment system were examined over a 26-mo period. Incidence of the protozoans Cryptosporidium spp. and Giardia spp. was also assessed. The control system utilizes a shallow basin to collect liquid runoff and accumulate eroded solids from the pen surfaces; when an adequate liquid volume is attained, the liquid is discharged from the basin onto a 4.5-ha vegetative treatment area (VTA) of bromegrass which is harvested as hay. Basin discharge transported E. coli O157, Campylobacter spp., and generic E. coli into the VTA soil, but without additional discharge from the basin, the pathogen prevalences decreased over time. Similarly, the VTA soil concentrations of generic E. coli initially decreased rapidly, but lower residual populations persisted. Isolation of Cryptosporidium oocysts and Giardia cysts from VTA samples was infrequent, indicating differences in sedimentation and/or transport in comparison to bacteria. Isolation of generic E. coli from freshly cut hay from VTA regions that received basin discharge (12 of 30 vs. 1 of 30 control samples) provided evidence for the risk of contamination; however, neither E. coli O157 or Campylobacter spp. were recovered from the hay following baling. This work indicates that the runoff control system is effective for reducing environmental risk by containing and removing pathogens from feedlot runoff.     

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。     

The microbial status of natural waters in a protected wilderness area

Waters derived from remote 'wilderness' locations have been assumed to be largely free of bacterial contamination and thus such, near-pristine, protected catchments, unused for agriculture, have been first in the multiple line of protection (pristine catchment-long storage-treatment-disinfection) employed by the water industry. This assumption is challenged by a bacterial survey of the waters derived from the New Cairngorm National Park, Scotland. Over 480 spot samples were taken for 59 sites between March 2001 and October 2002 during nine field campaigns each of three to five days duration. Over 75% of samples tested positive for Escherichia coli (E. coli) and 85% for total coliforms. Concentrations displayed both temporal and spatial patterns. Largest values occurred over the summer months and particularly at weekends at sites frequented by visitors, either for 'wild' camping or day visits, or where water was drawn from the river for drinking. Overall the spatial and temporal variations in bacterial concentrations suggest a relationship with visitor numbers and in particular wild camping. The implications of the results for drinking water quality and visitors health are discussed along with possible management options for the area in terms of improving the disposal of human waste.