Antibiotic resistance profiles to determine sources of fecal contamination in a rural Virginia watershed

Antibiotic resistance analysis (ARA) was used to determine if enterococci of human origin were present in a stream (Spout Run) that passes through a rural nonsewered community (Millwood, VA). Millwood consists of 82 homes, all served by individual septic systems, and Spout Run drains a 5,800-ha karst topography watershed that contains large populations of livestock and wildlife. Periodic monitoring by state regulatory officials had resulted in Spout Run being placed on the Virginia impaired stream list and Millwood being categorized as an at-risk community. Stream samples were collected monthly and analyzed for fecal coliforms and enterococci (May 1999-May 2000); ARA was performed on enterococci stream isolates on a quarterly basis. All 117 stream samples were positive for fecal coliforms, and 32% exceeded the Virginia recreational water standard (1,000 fecal coliforms/100 mL). A library of 1,174 known source Enterococcus isolate antibiotic resistance profiles was constructed, and yielded correct classification rates of 94.6% for 203 human isolates, 93.7% for 734 livestock isolates, and 87.8% for 237 wildlife isolates. Antibiotic resistance analysis of 2,012 enterococcal isolates recovered from stream samples indicated isolates of human origin appeared throughout the stream as it passed through Millwood, with a yearly average of approximately 10% human, 40% wildlife, and 50% livestock. There were no human origin isolates in samples collected upstream from Millwood, and the percent human origin isolates declined downstream from Millwood. While a human signature was found in Spout Run, it was small compared with the proportion of isolates from livestock and wildlife.     

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.     

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.     

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.     

Identifying sources of fecal contamination inexpensively with targeted sampling and bacterial source tracking

Most bacterial source tracking (BST) methods are too expensive for most communities to afford. We developed targeted sampling as a prelude to BST to reduce these costs. We combined targeted sampling with three inexpensive BST methods, Enterococcus speciation, detection of the esp gene, and fluorometry, to confirm the sources of fecal contamination to beaches on Georgia's Jekyll and Sea Islands during calm and stormy weather conditions. For Jekyll Island, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. For the Sea Island beach during calm conditions, the most likely sources of fecal contamination were leaking sewer lines and wildlife feces. The leaking sewer lines were confirmed with fluorometry and detection of the esp gene. For the Sea Island beach during stormflow conditions, the most likely sources of fecal contamination were wildlife feces and runoff discharging from two county-maintained pipes. For the pipes, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. Sediments were also a reservoir of fecal enterococci for both Jekyll and Sea Islands. Combining targeted sampling with two or more BST methods identified sources of fecal contamination quickly, easily, and inexpensively. This combination was the first time targeted sampling was conducted during stormy conditions, and the first time targeted sampling was combined with enterococcal speciation, detection of the esp gene, and fluorometry.     

Geographic variability of Escherichia coli ribotypes from animals in Idaho and Georgia

Several genotypic methods have been developed for determining the host origin of fecal bacteria in contaminated waters. Some of these methods rely on a host origin database to identify environmental isolates. It is not well understood to what degree these host origin isolates are geographically variable (i.e., cosmopolitan or endemic). This is important because a geographically limited host origin database may or may not be universally applicable. The objective of our study was to use one genotypic method, ribotyping, to determine the geographic variability of the fecal bacterium, Escherichia coli, from one location in Idaho and three locations in Georgia for cattle (Bos taurus), horse (Equus caballus), swine (Sus scrofa), and chicken (Gallus gallus domesticus). A total of 568 fecal E. coli isolates from Kimberly, ID (125 isolates), Athens, GA (210 isolates), Brunswick, GA (102 isolates), and Tifton, GA (131 isolates), yielded 213 ribotypes. The percentage of ribotype sharing within an animal species increased with decreased distance between geographic locations for cattle and horses, but not for swine and chicken. When the E. coli ribotypes among the four host species were compared at one location, the percent of unshared ribotypes was 86, 89, 81, and 79% for Kimberly, Athens, Brunswick, and Tifton, respectively. These data suggest that there is good ribotype separation among host animal species at each location. The ability to match environmental isolates to a host origin database may depend on a large number of environmental and host origin isolates that ideally are not geographically separated.     

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.     

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.     

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.     

ENHANCING FECAL COLIFORM TOTAL MAXIMUM DAILY LOAD MODELS THROUGH BACTERIAL SOURCE TRACKING1

ABSTRACT: Surface water impairment by fecal coliform bacteria is a water quality issue of national scope and importance. In Virginia, more than 400 stream and river segments are on the Commonwealth's 2002 303(d) list because of fecal coliform impairment. Total maximum daily loads (TMDLs) will be developed for most of these listed streams and rivers. Information regarding the major fecal coliform sources that impair surface water quality would enhance the development of effective watershed models and improve TMDLs. Bacterial source tracking (BST) is a recently developed technology for identifying the sources of fecal coliform bacteria and it may be helpful in generating improved TMDLs. Bacterial source tracking was performed, watershed models were developed, and TMDLs were prepared for three streams (Accotink Creek, Christians Creek, and Blacks Run) on Virginia's 303(d) list of impaired waters. Quality assurance of the BST work suggests that these data adequately describe the bacteria sources that are impairing these streams. Initial comparison of simulated bacterial sources with the observed BST data indicated that the fecal coliform sources were represented inaccurately in the initial model simulation. Revised model simulations (based on BST data) appeared to provide a better representation of the sources of fecal coliform bacteria in these three streams. The coupled approach of incorporating BST data into the fecal coliform transport model appears to reduce model uncertainty and should result in an improved TMDL.     

Modeling Flow,Nutrient, and Sediment Delivery from a Large International Watershed Using a Field‐Scale SWAT Model

A large international watershed, the St. Clair‐Detroit River System, containing both extensive urban and agricultural areas, was modeled using the Soil and Water Assessment Tool (SWAT) model. The watershed, located in southeastern Michigan, United States, and southwestern Ontario, Canada, encompasses the St. Clair, Clinton, Detroit (DT), Sydenham (SY), Upper, and Lower Thames subwatersheds. The SWAT input data and model resolution (i.e., hydrologic response units, HRUs), were established to mimic farm boundaries, the first time this has been done for a watershed of this size. The model was calibrated (2007–2015) and validated (2001–2006) with a mix of manual and automatic methods at six locations for flow and water quality at various time scales. The model was evaluated using Nash–Sutcliffe efficiency and percent bias and was used to explore major water quality issues. We showed the importance of allowing key parameters to vary among subwatersheds to improve goodness of fit, and the resulting parameters were consistent with subwatershed characteristics. Agricultural sources in the Thames and SY subwatersheds and point sources from DT subwatershed were major contributors of phosphorus. Spatial distribution of phosphorus yields at HRU and subbasin levels identified locations for potential management targeting for both point and nonpoint sources and revealed that in some subwatersheds nonpoint sources are dominated by urban sources.     

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.     

Interaction and influence of two creeks on Escherichia coli concentrations of nearby beaches: exploration of predictability and mechanisms

The impact of river outfalls on beach water quality depends on numerous interacting factors. The delivery of contaminants by multiple creeks greatly complicates understanding of the source contributions, especially when pollution might originate up- or down-coast of beaches. We studied two beaches along Lake Michigan that are located between two creek outfalls to determine the hydrometeorologic factors influencing near-shore microbiologic water quality and the relative impact of the creeks. The creeks continuously delivered water with high concentrations of Escherichia coli to Lake Michigan, and the direction of transport of these bacteria was affected by current direction. Current direction reversals were associated with elevated E. coli concentrations at Central Avenue beach. Rainfall, barometric pressure, wave height, wave period, and creek specific conductance were significantly related to E. coli concentration at the beaches and were the parameters used in predictive models that best described E. coli variation at the two beaches. Multiple inputs to numerous beaches complicates the analysis and understanding of the relative relationship of sources but affords opportunities for showing how these complex creek inputs might interact to yield collective or individual effects on beach water quality.     

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.     

Respective contributions of point and non-point sources of E. coli and enterococci in a large urbanized watershed (the Seine river, France)

Because the large rivers of the Seine watershed have a low microbiological water quality, the main sources of fecal contamination were investigated in the present study. The inputs of the point (wastewater treatment plants (WWTPs) effluents) and non-point sources (surface runoff and soil leaching) of fecal bacteria were quantified for Escherichia coli and intestinal enteroccoci used as bacterial indicators. In order to assess the contamination through non-point sources, fecal indicators abundance was estimated in samples collected in small streams located in rural areas upstream from all point sources; these small rivers were characterized by the land use of their watershed. Bacterial indicator numbers were also measured in effluents of WWTPs, some using classical treatment (settling followed by activated sludge process) and some using an additional disinfection stage (UV irradiation). These data were used to estimate the respective importance of each type of source at the scale of the whole Seine river watershed taking into account the land use and the population density. It shows the predominant importance of the point sources of fecal indicator bacteria at the scale of the whole watershed. In a scenario in which activated sludge treatment would be complemented with UV in all WWTPs located in this watershed, the non-point sources of fecal indicator bacteria would be dominant.     

Waterfowl abundance does not predict the dominant avian source of beach Escherichia coli

The horizontal, fluorophore enhanced, rep-PCR (HFERP) DNA fingerprinting technique was used to identify potential sources of in water, nearshore sand, and sediment at two beaches in the Duluth-Superior Harbor, near Duluth, MN, and Superior, WI, during May, July, and September 2006. An animal or environmental source could be identified for 35, 29, and 30% of strains in water, sand, and sediments, respectively. Waterfowl, including Canada geese, ring-billed gulls, and mallard ducks, were the largest source of that could be identified in water (55-100%), sand (59-100%), and sediment (92-100%) at both beaches. Although ring-billed gulls were more abundant in this harbor, Canada geese were usually the dominant source of waterfowl found at these beaches. The percentage of identified from treated wastewater was always less than the percentage of originating from waterfowl. At both beaches, the percentage of in water contributed by treated wastewater was higher in May compared with July and September. The larger proportion of wastewater-derived seen in May probably reflected a smaller contribution of from geese when these birds were less abundant rather than an absolute increase in from treated wastewater. Microbial source analysis and bird census data both indicated that waterfowl were a major source of at beaches in the Duluth-Superior Harbor. These data also indicated it is risky to assume that the most abundant waterfowl species present in waterways will also be the largest source of avian-derived in water, nearshore sand, and sediments at beaches.     

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.     

Use of Escherichia coli BOX-PCR fingerprints to identify sources of fecal contamination of water bodies in the State of São Paulo, Brazil

Repetitive element sequence-based polymerase chain reaction (rep-PCR) is one of the commonest methods used to identify sources of fecal contamination of water systems. In this work, BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR) was used to discriminate Escherichia coli strains originating from different animals and water sources, and the suitability of the technique for bacterial source tracking (BST) was evaluated. A total of 214 strains from humans, 150 strains from animals, 55 strains from sewage and 77 strains from water bodies were analyzed by the BOX-PCR technique. When maximum similarity between the fingerprints was used, a correct classification rate of 84% was achieved for strains from human and animal sources. Furthermore, 95% of the strains found in sewage were classified as being from human sources by at least one of the four classification tools used. Classification of the strains found in water bodies in the State of S?o Paulo was based on the fingerprints obtained for human and animal sources. Most of the sampling sites appeared to be affected by mixed sources of fecal contamination. The use of BOX-PCR for BST could be especially valuable in developing countries, where simplicity and cost are important considerations.     

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.     

Potential of Enterococcus faecalis as a human fecal indicator for microbial source tracking

Regulatory agencies are interested in a fecal indicator bacterium with a host range limited to humans because human fecal contamination represents the greatest hazard to humans, yet is a relatively easy nonpoint source to remedy. Watersheds with human fecal contamination could be given first priority for cleanup. A fecal indicator bacterium with a host range limited to humans and a few other warm-blooded animal species would also simplify microbial source tracking because only a few animal species would be required for any host origin database. The literature suggests that the fecal indicator bacterium Enterococcus faecalis has a limited host range. On this basis, we selected this bacterium for study. Of 583 fecal streptococcal isolates obtained on Enterococcosel agar from Canada goose, cattle, deer, dog, human, chicken, and swine, 392 were considered presumptive enterococci and were subsequently speciated with the API 20 Strep system. Of these isolates, 22 were Ent. durans (5.6%), 61 were Ent. faecalis (15.6%), 98 were Ent. faecium (25.0%), 86 were Ent. gallinarum (21.9%), and 125 were unidentified (31.9%). The host range of the Ent. faecalis isolates was limited to dogs, humans, and chickens. Media were developed to isolate and identify Ent. faecalis quickly from fecal samples and this scheme eliminated Ent. faecalis isolates from dogs. When the remaining Ent. faecalis isolates were ribotyped, it was possible to differentiate clearly among the isolates from human and chicken. It may be that combining the potentially limited host range of Ent. faecalis with ribotyping is useful for prioritizing watersheds with fecal contamination.