Modeling the dry-weather tidal cycling of fecal indicator bacteria in surface waters of an intertidal wetland

Recreational water quality at beaches in California and elsewhere is often poor near the outlets of rivers, estuaries, and lagoons. This condition has prompted interest in the role of wetlands in modulating surface water concentrations of fecal indicator bacteria (FIB), the basis of water quality standards internationally. A model was developed and applied to predict the dry-weather tidal cycling of FIB in Talbert Marsh, an estuarine, intertidal wetland in Huntington Beach, California, in response to loads from urban runoff, bird feces, and resuspended sediments. The model predicts the advection, dispersion and die-off of total coliform, Escherichia coli, and enterococci using a depth-integrated formulation. We find that urban runoff and resuspension of contaminated wetland sediments are responsible for surface water concentrations of FIB in the wetland. Model predictions show that urban runoff controls surface water concentrations at inland sites and sediment resuspension controls surface water concentrations near the mouth. Direct wash-off of bird feces into the surface water is not a significant contributor, although bird feces can contribute to the sediment bacteria load. The key parameters needed to accurately predict FIB concentrations, using a validated hydrodynamic model, are: the load due to urban runoff, sediment erodibility parameters, and sediment concentrations and surface water die-off rates of enteric bacteria. In the present study, literature values for sediment erodibility and water column die-off rates are used and average concentrations of FIB are predicted within 1/2 log unit of measurements. Total coliform are predicted more accurately than E. coli or enterococci, both in terms of magnitude and tidal variability. Since wetland-dependent animals are natural sources of FIB, and FIB survive for long periods of time and may multiply in wetland sediments, these results highlight limitations of FIB as indicators of human fecal pollution in and near wetlands.     

Variability of fecal indicator bacteria in flowing and ponded waters in southern California: implications for bacterial TMDL development and implementation

Recreational water quality is assessed by using water quality objectives for fecal indicator bacteria (FIB) including total coliform, fecal coliform (or E. coli), and/or Enterococcus. It is required under the Clean Water Act that a TMDL be developed for a bacteria-impaired water body. The development and implementation of bacterial TMDLs has proven challenging and often difficult due to unknown source(s) of FIB. This study found that FIB levels varied significantly in flowing water, ponded water, and associated sediment. FIB levels in isolated ponded water in waterways were significantly higher than in flowing water. Sediment under ponded water contained a great amount of FIB. Furthermore, FIB concentrations in ponded water tended to increase with increasing water temperature and to decrease with increasing water salinity. The result provides the field evidence of survival/growth of FIB in water and sediment under ambient conditions in southern California. A holistic approach including natural sources (e.g., a reference system) should be considered for practical and applicable purposes while developing and implementing bacterial TMDLs for pathogen-impaired waterbodies.     

Addressing uncertainty in fecal indicator bacteria dark inactivation rates

Assessing the potential threat of fecal contamination in surface water often depends on model forecasts which assume that fecal indicator bacteria (FIB, a proxy for the concentration of pathogens found in fecal contamination from warm-blooded animals) are lost or removed from the water column at a certain rate (often referred to as an “inactivation” rate). In efforts to reduce human health risks in these water bodies, regulators enforce limits on easily-measured FIB concentrations, commonly reported as most probable number (MPN) and colony forming unit (CFU) values. Accurate assessment of the potential threat of fecal contamination, therefore, depends on propagating uncertainty surrounding “true” FIB concentrations into MPN and CFU values, inactivation rates, model forecasts, and management decisions. Here, we explore how empirical relationships between FIB inactivation rates and extrinsic factors might vary depending on how uncertainty in MPN values is expressed. Using water samples collected from the Neuse River Estuary (NRE) in eastern North Carolina, we compare Escherichia coli (EC) and Enterococcus (ENT) dark inactivation rates derived from two statistical models of first-order loss; a conventional model employing ordinary least-squares (OLS) regression with MPN values, and a novel Bayesian model utilizing the pattern of positive wells in an IDEXX Quanti-Tray^®/2000 test. While our results suggest that EC dark inactivation rates tend to decrease as initial EC concentrations decrease and that ENT dark inactivation rates are relatively consistent across different ENT concentrations, we find these relationships depend upon model selection and model calibration procedures. We also find that our proposed Bayesian model provides a more defensible approach to quantifying uncertainty in microbiological assessments of water quality than the conventional MPN-based model, and that our proposed model represents a new strategy for developing robust relationships between environmental factors and FIB inactivation rates, and for reducing uncertainty in water resource management decisions.     

Persistence of fecal indicator bacteria in Santa Monica Bay beach sediments

Monitoring the water quality of recreational beaches is only one step toward understanding microbial contamination -- the primary cause of beach closings. The surf zone sediment reservoir is typically overlooked and may also be important. This study involved monitoring the fecal indicator bacteria (FIB) levels in water and sediment at three ocean beaches (two exposed and one enclosed) during a storm event, conducting laboratory microcosm experiments with sediment from these beaches, and surveying sediment FIB levels at 13 beaches (some exposed and some enclosed). Peaks in Escherichia coli and enterococci concentrations in water and sediment coincided with storm activity at the two exposed beaches, while levels of both FIB were consistently high and irregular at the enclosed beach. Results from microcosm experiments showing similar, dramatic growth of FIB in both overlying water and sediment from all beaches, as well as results from the beach survey, support the hypothesis that the quiescent environment rather than sediment characteristics can explain the elevated sediment FIB levels observed at enclosed beaches. This work has implications for the predictive value of FIB measurements, and points to the importance of the sediment reservoir.     

Meteorological effects on the levels of fecal indicator bacteria in an urban stream: A modeling approach

Gwangju Creek (GJC) in Korea, which drains a highly urbanized watershed, has suffered from substantial fecal contamination, thereby limiting the beneficial use of the water in addition to threatening public health. In this study, to quantitatively estimate the sinks and sources of fecal indicator bacteria (FIB) in GJC under varying meteorological conditions, two FIB (i.e., Escherichia coli and enterococci bacteria) were monitored hourly for 24 h periods during both wet and dry weather conditions at four sites along GJC, and the collected data was subsequently used to develop a spatiotemporal FIB prediction model. The monitoring data revealed that storm washoff and irradiational die-off by sunlight are the two key processes controlling FIB populations in wet and dry weather, respectively. FIB populations significantly increased during precipitation, with greater concentrations occurring at higher rainfall intensity. During dry weather, FIB populations decreased in the presence of sunlight in daytime but quickly recovered at nighttime due to continuous point-source inputs. In this way, the contributions of the key processes (i.e., irradiational die-off by sunlight, settling, storm washoff, and resuspension) to the FIB levels in GJC under different meteorological conditions were quantitatively estimated using the developed model. The modeling results showed that the die-off by sunlight is the major sink of FIB during the daytime in dry weather with a minor contribution from the settling process. During wet weather, storm washoff and resuspension are equally important processes that are responsible for the substantial increase of FIB populations.     

Use of salinity mixing models to estimate the contribution of creek water fecal indicator bacteria to an estuarine environment: Newport Bay, California

The contribution of freshwater discharge to fecal indicator bacteria (FIB) impairment of an estuarine environment can be approximated from simple, two end-member mixing models using salinity as a tracer. We conducted a yearlong time series investigation of Newport Bay, a regionally important estuarine embayment in southern California, assessing the concentrations of FIB, specifically Escherichia coli and enterococci bacteria, and salinity. In total, eight within-bay stations and one offshore control site were sampled nearly once per week and the three tributaries draining into Newport Bay were sampled approximately daily. Using salinity as a conservative tracer for water mass mixing and determining the end-member values of FIB in both the creek sites and the offshore site, we created a linear, two end-member mixing model of FIB within Newport Bay. Deviations from the mixing model suggest either an additional source of FIB to the bay (e.g. bird feces, storm drain discharge) or regrowth and/or die-off of FIB within the bay. Our results indicate that salinity mixing models can be useful in predicting changes in FIB concentrations in the estuarine environments and can help narrow the search for sources of FIB to the bay and enhance our understanding of the fate of FIB within the bay.     

Assessment of sources of human pathogens and fecal contamination in a Florida freshwater lake

We investigated the potential for a variety of environmental reservoirs to harbor or contribute fecal indicator bacteria (FIB), DNA markers of human fecal contamination, and human pathogens to a freshwater lake. We hypothesized that submerged aquatic vegetation (SAV), sediments, and stormwater act as reservoirs and/or provide inputs of FIB and human pathogens to this inland water. Analysis included microbial source tracking (MST) markers of sewage contamination (Enterococcus faecium esp gene, human-associated Bacteroides HF183, and human polyomaviruses), pathogens (Salmonella, Cryptosporidium, Giardia, and enteric viruses), and FIB (fecal coliforms, Escherichia coli, and enterococci). Bayesian analysis was used to assess relationships among microbial and physicochemical variables. FIB in the water were correlated with concentrations in SAV and sediment. Furthermore, the correlation of antecedent rainfall and major rain events with FIB concentrations and detection of human markers and pathogens points toward multiple reservoirs for microbial contaminants in this system. Although pathogens and human-source markers were detected in 55% and 21% of samples, respectively, markers rarely coincided with pathogen detection. Bayesian analysis revealed that low concentrations (<45 CFU × 100 ml⁻¹) of fecal coliforms were associated with 93% probability that pathogens would not be detected; furthermore the Bayes net model showed associations between elevated temperature and rainfall with fecal coliform and enterococci concentrations, but not E. coli. These data indicate that many under-studied matrices (e.g. SAV, sediment, stormwater) are important reservoirs for FIB and potentially human pathogens and demonstrate the usefulness of Bayes net analysis for water quality assessment.     

Human and bovine adenoviruses for the detection of source-specific fecal pollution in coastal waters in Australia

In this study, the host-specificity and -sensitivity of human- and bovine-specific adenoviruses (HS-AVs and BS-AVs) were evaluated by testing wastewater/fecal samples from various animal species in Southeast, Queensland, Australia. The overall specificity and sensitivity of the HS-AVs marker were 1.0 and 0.78, respectively. These figures for the BS-AVs were 1.0 and 0.73, respectively. Twenty environmental water samples were collected during wet conditions and 20 samples were colleted during dry conditions from the Maroochy Coastal River and tested for the presence of fecal indicator bacteria (FIB), host-specific viral markers, zoonotic bacterial and protozoan pathogens using PCR/qPCR. The concentrations of FIB in water samples collected after wet conditions were generally higher compared to dry conditions. HS-AVs was detected in 20% water samples collected during wet conditions and whereas BS-AVs was detected in both wet (i.e., 10%) and dry (i.e., 10%) conditions. Both Campylobacter jejuni mapA and Salmonella invA genes detected in 10% samples collected during dry conditions. The concentrations of Salmonella invA ranged between 3.5 × 10² and 4.3 × 10² genomic copies per 500 ml of water Giardia lamblia β-giardin gene was detected only in one sample (5%) collected during the dry conditions. Weak or significant correlations were observed between FIB with viral markers and zoonotic pathogens. However, during dry conditions, no significant correlations were observed between FIB concentrations with viral markers and zoonotic pathogens. The prevalence of HS-AVs in samples collected from the study river suggests that the quality of water is affected by human fecal pollution and as well as bovine fecal pollution. The results suggest that HS-AVs and BS-AVs detection using PCR could be a useful tool for the identification of human sourced fecal pollution in coastal waters.     

A mechanistic model of runoff-associated fecal coliform fate and transport through a coastal lagoon

Fecal coliform (FC) contamination in coastal waters is an ongoing public health problem worldwide. Coastal wetlands and lagoons are typically expected to protect coastal waters by attenuating watershed pollutants including FC bacteria. However, new evidence suggests that coastal lagoons or marshes can also be a source of high indicator organism concentrations in coastal waters. We asked for a Mediterranean-type climate, what is the fate of runoff-associated FC through a coastal lagoon? To address this question, we developed a mass balance-based, mechanistic model of FC concentration through a coastal lagoon and simulated, for summer and winter conditions, FC within the lagoon water column, lagoon sediments, and in the ocean water just downstream of the lagoon mouth. Our model accounts for advective flow and dispersion, decay and sedimentation and resuspension of FC-laden sediments during high flow, erosional conditions. Under low flow conditions that occur in the summer, net FC decay and FC storage in lagoon sediments are predicted. Under high flow conditions that occur in the winter, FC-laden sediments are predicted to erode, resuspend and flow out of the lagoon where they elevate FC concentrations in the coastal ocean. For both seasonal conditions, the predicted water column FC concentrations were within an order of magnitude of field measurements for a reference site in southern California. Our results suggest that there are seasonally varying roles for coastal lagoons in mediating FC contamination to coastal waters.     

Growing season surface water loading of fecal indicator organisms within a rural watershed

The loading of microbial contaminants was examined within the Thomas Brook watershed, a 784 ha mixed land-use catchment located in the headwaters of the Cornwallis River drainage basin (Nova Scotia, Canada). The objectives were to: (i) examine spatial and temporal characteristics of fecal bacteria loading during the growing season from five subwatersheds, and (ii) develop areal fecal indicator organism export coefficients for rural landscapes. Fecal coliform, Escherichia coli, total suspended solids (TSS) concentrations and stream flow were monitored at five locations in the watershed over six consecutive growing seasons (May-Oct, 2001-2006). A nested watershed monitoring approach was used to determine bacterial loading from distinct source types (residential vs. agricultural) during both baseflow and stormflow periods. Areal bacterial loading rates increased in each nested watershed moving downstream through the watershed and were highest in the three subcatchments dominated by agricultural activities. Upper watershed bacterial loading throughout the growing season from an agricultural subcatchment (Growing Season Avg 8.92 × 10¹⁰ CFU ha⁻¹) was consistently higher than a residential subcatchment (Growing Season Avg 8.43 × 10⁹ CFU ha⁻¹). As expected, annual average stormflow bacterial loads were higher than baseflow loads, however baseflow loads still comprised between 14 and 35% of the growing season bacterial loads in the five subwatersheds. Fecal bacteria loads were greater during years with higher annual precipitation. A positive linear relationship was observed between E. coli and TSS loading during the 2005 and 2006 growing seasons when both parameters were monitored, indicating that the processes of sediment transport and bacterial transport are linked. It is anticipated that computed areal microbial loading coefficients will be useful in developing watershed management plans. More intensive sampling during stormflow events is recommended for improving these coefficients.     

Reduction of fecal indicator bacteria (FIB) in the Ballona Wetlands saltwater marsh (Los Angeles County, California, USA) with implications for restoration actions

A benefit of wetland preservation and restoration is the ecosystem service of improving water quality, typically assessed based on bacterial loading. The Ballona Wetlands, a degraded salt marsh of approximately 100 ac located on the southern border of Marina Del Rey (Los Angeles County, California, USA) are currently the focus of publicly funded restoration planning. The wetlands receive tidal water, usually contaminated with fecal indicator bacteria (FIB: total and fecal coliforms, Escherichia coli, enterococci) from the adjacent Ballona Creek and Estuary. During the summer of 2007, two 24-h studies were conducted to determine FIB tidal dynamics within the wetland. Measurements of water flow and mean FIB concentrations (n = 3) were measured every 1.5 h to determine total FIB load estimates. FIB loading rates (MPN/s) were greatest during flood tides as water entered the wetlands, and then again during spring tide conditions when sediments were resuspended during swifter spring ebb flows. During daylight hours, the wetland acted as a sink for these bacteria as loads diminished, presumably by sunlight and other processes. Conversely, during late afternoon and night, the wetlands shifted to being a source as excess FIB departed on ebb flows. Therefore, the wetlands act as both a source and sink for FIB depending on tidal conditions and exposure to sunlight. Future restoration actions would result in a tradeoff - increased tidal channels offer a greater surface area for FIB inactivation, but also would result in a greater volume of FIB-contaminated resuspended sediments carried out of the wetlands on stronger ebb flows. As levels of FIB in Ballona Creek and Estuary diminish through recently established regulatory actions, the wetlands could shift into a greater sink for FIB.     

Characterizing fecal contamination in stormwater runoff in coastal North Carolina, USA

Microbial contaminants in stormwater runoff have the potential to negatively impact public health. Stormwater runoff to coastal waters is increasing in amount and rate of discharge due to loss of vegetated landscape and increasing coastal development. However, the extent and nature of microbial contamination of stormwater runoff in North Carolina (NC) has not been previously characterized. The aim of this study was to measure a range of fecal indicator bacteria (FIB) and molecular markers at three coastal sites. E. coli and Enterococcus sp. were measured in addition to molecular markers including Bacteroides Human-Specific Marker (HS) and fecal Bacteroides spp. Levels of FIB in stormwater far exceeded recreational water quality guidelines, frequently by several orders of magnitude. High concentrations of fecal Bacteroides spp. and the presence of HS indicated the presence of human fecal contamination in the stormwater runoff, but only during specific storms. Examinations of levels of fecal contamination in stormwater over multiple seasons and a range of storm conditions will allow managers to consider appropriate design of effective mitigation strategies necessary to maintain and restore coastal water quality.     

Contrasts in concentrations and loads of conventional and alternative indicators of fecal contamination in coastal stormwater

Fecal contamination in stormwater is often complex. Because conventional fecal indicator bacteria (FIB) cannot be used to ascertain source of fecal contamination, alternative indicators are being explored to partition these sources. As they are assessed for future use, it is critical to compare alternative indicators to conventional FIB under a range of stormwater delivery conditions. In this study, conventional FIB and fecal Bacteroides spp. were monitored throughout the duration of five storm events from coastal stormwater outfalls in Dare County, North Carolina, USA to characterize relationships among FIB concentrations, alternative fecal markers, and loading of contaminants. Water samples were collected multiple times during each storm and analyzed for Enterococcus sp. and Escherichia coli using enzymatic tests and fecal Bacteroides spp. by QPCR. Both conventional FIB and fecal Bacteroides spp. concentrations in stormwater were generally high and extremely variable over the course of the storm events. Over the very short distances between sites, we observed statistically significant spatial and temporal variability, indicating that stormwater monitoring based on single grab-samples is inappropriate. Loading of FIB and fecal Bacteroides spp. appeared to be affected differently by various hydrologic factors. Specifically, Spearman correlations between fecal Bacteroides spp. and drainage area and antecedent rainfall were lower than those between conventional FIB and these hydrologic factors. Furthermore, the patterns of fecal Bacteroides spp. concentrations generally increased over the duration of the storms, whereas E. coli and Enterococcus sp. concentrations generally followed the patterns of the hydrograph, peaking early and tailing off. Given the greater source-specificity and limited persistence of fecal Bacteroides spp. in oxygenated environments, differences in these patterns suggest multiple delivery modes of fecal contamination (i.e. landscape scouring versus groundwater discharge).     

Evaluation of rapid methods and novel indicators for assessing microbiological beach water quality

A broad suite of new measurement methods and indicators based on molecular measurement technology have been developed to assess beach water quality, but they have generally been subjected to limited testing outside of the laboratory in which they were developed. Here we evaluated 29 assays targeting a variety of bacterial, viral, and chemical analytes by providing the method developers with twelve blind samples consisting of samples spiked with known concentration of sewage or gull guano and negative controls. Each method was evaluated with respect to its ability to detect the target organism, absence of signal in the negative controls and repeatability among replicates. Only six of the 30 methods detected their targets in at least 75% of the samples while consistently determining the absence of the target in the negative controls. Among quantitative methods, QPCR for Bacteroides thetaiotamicron and Enterococcus detected by Luminex reliably identified all but one sample containing human fecal material and produced no false positive results. Among non-quantitative methods, the Enterococcus esp gene, the Bacteroidales human specific marker and culture-based coliphage were the most reliable for identifying human fecal material. We also found that investigator-specific variations of methods targeting the same organism often produced different results.     

Loading of fecal indicator bacteria in North Carolina tidal creek headwaters: Hydrographic patterns and terrestrial runoff relationships

In the New River Estuary (NRE) in eastern North Carolina (NC), fecal indicator bacteria (FIB) levels exceed water quality standards, leading to closure of estuarine waters for shellfishing and classification of parts of the estuary as “impaired” per the Clean Water Act section 303(d) list. As a means to investigate fecal contamination and loading of FIB to the NRE, a continuous automated sampler (ISCO) outfitted with flow modules and water quality probes was placed in four first-order tidal creek headwaters. Total storm discharge and bacterial load for Escherichia coli (EC) and Enterococcus spp. (ENT) were calculated using graphical volumetric flow calculations and interpolation of FIB measurements over each storm’s duration for 10 storms. Mean total load of 10⁹- 10¹² EC and ENT cells (MPN) occurred over the course of each storm. Total storm loading, averaged across all storms, was as much as 30 and 37 times greater than equivalent duration of baseflow loading for EC and ENT, respectively. Within the first 30% of creek storm volume for all storms and all creeks combined, a mean cumulative load of only 37% and 44% of the total EC and ENT cells, respectively, was discharged, indicating these creeks are not demonstrating a ‘first flush’ scenario for FIB. The median storm Event Mean Concentrations (EMCs) were 6.37 × 10² and 2.03 × 10² MPN/100 mL, for EC and ENT, respectively, compared with median baseflow concentrations of 1.48 × 10² and 4.84 × 10¹ for EC and ENT, respectively, and were significantly different between base and storm flow events. FIB was correlated with TSS (weak), flow rate (strong), and different stages (base, rising, peak, and falling) of the hydrograph (strong). Pollutographs indicate large intra-storm variability of FIB, and the need for more intensive sampling throughout a storm in order to attain accurate FIB contaminant estimates. Instream sediment concentrations ranged from 5 to 478 (MPN/g) and 13 to 776 (MPN/g) for EC and ENT, respectively, indicating sediment as a source, but a minor reservoir. This overall approach for calculating loading in headwater tidal creeks is detailed. Accurate loading characterization of FIB during storms and dry weather conditions, and understanding intra-storm FIB concentrations, is imperative for understanding patterns of water quality impairment, establishing management planning, and developing appropriate mitigation strategies.     

Presence of Enterococcus faecalis in broiler litter and wild bird feces for bacterial source tracking

When Enterococcus faecalis is isolated from fresh feces, its host range appears to be limited to humans and birds. Although E. faecalis is found in human sewage, the extent to which the bacterium is found in broiler litter and in the feces of wild birds is unclear. These results have implications for bacterial source tracking. We determined if media designed for the isolation of fecal enterococci affected this host range, and if E. faecalis was routinely found in broiler litter and in the feces of wild birds. Of five different isolation media, none affected the isolation of E. faecalis. Enterococcus faecalis was routinely found in fresh broiler feces (522 of 1092 isolates; 48%), but rarely in broiler litter (12 of 1452 isolates; <2%). Therefore, broiler litter selects against this bacterium, and broiler litter is an unlikely environmental source of this bacterium. The presence of E. faecalis in eight wild bird species was highly variable. Unless the fecal loading rate from migratory or resident wild birds is high, water samples collected during baseflow conditions with high numbers of E. faecalis may indicate human fecal contamination.     

Survival dynamics of fecal bacteria in ponds in agricultural watersheds of the Piedmont and Coastal Plain of Georgia

Animal agriculture in watersheds produces manure bacteria that may contaminate surface waters and put public health at risk. We measured fecal indicator bacteria (commensal Escherichia coli and fecal enterococci) and manure pathogens (Salmonella and E. coli 0157:H7), and physical-chemical parameters in pond inflow, within pond, pond outflow, and pond sediments in three ponds in agricultural watersheds. Bishop Pond with perennial inflow and outflow is located in the Piedmont, and Ponds A and C with ephemeral inflow and outflow in the Coastal Plain of Georgia. Bromide and chloride tracer experiments at Bishop Pond reflected a residence time much greater than that estimated by two models, and indicated that complete mixing within Bishop Pond was never obtained. The long residence time meant that fecal bacteria were exposed to solar UV-radiation and microbial predation. At Bishop Pond outflow concentrations of fecal indicator bacteria were significantly less than inflow concentrations; such was not observed at Ponds A and C. Both Salmonella and E. coli 0157:H7 were measured when concomitant concentrations of commensal E. coli were below the criterion for surface water impairment indicating problems with the effectiveness of indicator organisms. Bishop Pond improved down stream water quality; whereas, Ponds A and C with ephemeral inflow and outflow and possibly greater nutrient concentrations within the two ponds appeared to be less effective in improving down stream water quality.     

Rapid QPCR-based assay for fecal Bacteroides spp. as a tool for assessing fecal contamination in recreational waters

Concentrations of fecal indicator bacteria (FIB; e.g. Escherichia coli, and Enterococcus sp.) can only be used in limited ways for determining the source of fecal contamination in recreational waters because they cannot distinguish human from non-human fecal contamination. Several Bacteroides spp. have been suggested as potential alternative indicators. We have developed a rapid, culture-independent method for quantifying fecal Bacteroides spp. using quantitative PCR (QPCR) targeting the 16S rRNA gene. The assay specifically targets and quantifies the most common human Bacteroides spp. The details of the method are presented, including analyses of a wide range of fecal samples from different organisms. Specificity and performance of the QPCR assay were also tested via a laboratory experiment where human sewage and gull guano were inoculated into a range of environmental water samples. Concentrations of fecal Bacteroides spp., total Enterococcus sp., Enterococcus faecium, Enterococcus faecalis, and Enterococcus casseliflavus were measured using QPCR, and total Enterococcus sp. and E. coli were quantified by membrane filtration (MF). Samples spiked with gull guano were highly concentrated with total Enterococcus sp., E. coli, E. faecalis, and E. casseliflavus, demonstrating that these indicators are prominent in animal feces. On the other hand, fecal Bacteroides spp. concentrations were high in samples containing sewage and were relatively low in samples spiked with gull guano. Sensitivity and specificity results suggest that the rapid fecal Bacteroides spp. QPCR assay may be a useful tool to effectively predict the presence and concentration of human-specific fecal pollution.     

Fecal indicator bacteria and Salmonella in ponds managed as bird habitat, San Francisco Bay, California, USA

Throughout the world, coastal resource managers are encouraging the restoration of previously modified coastal habitats back into wetlands and managed ponds for their ecosystem value. Because many coastal wetlands are adjacent to urban centers and waters used for human recreation, it is important to understand how wildlife can affect water quality. We measured fecal indicator bacteria (FIB) concentrations, presence/absence of Salmonella, bird abundance, and physico-chemical parameters in two coastal, managed ponds and adjacent sloughs for 4 weeks during the summer and winter in 2006. We characterized the microbial water quality in these waters relative to state water-quality standards and examined the relationship between FIB, bird abundance, and physico-chemical parameters. A box model approach was utilized to determine the net source or sink of FIB in the ponds during the study periods. FIB concentrations often exceeded state standards, particularly in the summer, and microbial water quality in the sloughs was generally lower than in ponds during both seasons. Specifically, the inflow of water from the sloughs to the ponds during the summer, more so than waterfowl use, appeared to increase the FIB concentrations in the ponds. The box model results suggested that the ponds served as net wetland sources and sinks for FIB, and high bird abundances in the winter likely contributed to net winter source terms for two of the three FIB in both ponds. Eight serovars of the human pathogen Salmonella were isolated from slough and pond waters, although the source of the pathogen to these wetlands was not identified. Thus, it appeared that factors other than bird abundance were most important in modulating FIB concentrations in these ponds.     

Mortality rates of autochthonous and fecal bacteria in natural aquatic ecosystems

Bacterial mortality has been investigated in freshwater (River Seine) and in marine (North Sea) systems using a method based on following the disappearance of radioactivity from the DNA of assemblages of bacteria previously labeled with tritiated thymidine. Measurement of bacterial mortality of autochthonous and various types of fecal bacteria allowed direct comparisons between their respective first-order mortality rates. Mortality rates obtained for the different types of bacteria in the River Seine were, respectively, 7.9-33.9 x 10(-3)h(-1) for Escherichia coli, 12.2-29.2 x 10(-3)h(-1) for S. faecium and 7.0-18.3 x 10(-3)h(-1) for the autochthonous bacteria. In the Belgian coastal waters, these rates were 4.6-27.3 x 10(-3)h(-1) for E. coli, 6.0-22.0 x 10(-3)h(-1) for S. typhimurium, 10.0-18.9 x 10(-3)h(-1) for S. faecium and 1.0-13.9 x 10(-3)h(-1) for autochthonous bacteria. In both environments, the overall mortality rates of autochthonous and the different fecal bacteria were in the same order of magnitude and overall mortality rates of E. coli were on average about twice as high for autochthonous bacteria. Grazing by protozooplankton was the dominant process of mortality for fecal and autochthonous bacteria in both environments. Except in a few situations, grazing by protozooplankton was responsible for more than 90% of the overall mortality rate of fecal and autochthonous bacteria in the river and in the coastal area.