Fecal Indicator Bacteria Concentrations Research Articles

Consideration of Natural Sources in a Bacteria TMDL-Lines of Evidence, Including Beach Microbial Source Tracking.

Total Maximum Daily Load (TMDL) stipulations remained unmet at a southern California beach despite a suite of management actions carried out since 2001, prompting exploration of a Natural Sources Exclusion (NSE) provision within the TMDL. Quantitative Microbial Source Tracking (MST) was employed from 2012 to 2015 to inventory sources of natural and anthropogenic fecal indicator bacteria (FIB). Data suggested FIB exceedances could be traced to gulls based on gull marker prevalence and correlations with FIB concentrations in seawater, sand, and eelgrass. In contrast, human marker concentrations and a tracer dye test did not indicate prevalent human sources. Exponential decay of gull marker in sand amended with live Catellicoccus marimammalium suggested that measured marker reflected fecal inputs versus growth outside the host. Improved water quality was coincident with a 2013 bird exclusion structure, consistent with NSE. However, load allocation needed for TMDL reconsideration was hampered by variable ratios of FIB, MST markers, and pathogens measured in seawater and in gull, cat, and raccoon feces. Quantitative Microbial Risk Assessment is a suggested path forward because such models can incorporate distributions from a combination of FIB sources and communicate criteria in terms of human health risk.

Nutrients and Phytoplankton in a Shallow, Hypereutrophic Urban Lake: Prospects for Restoration

University Lake, a shallow, artificial, urban lake adjacent to the campus of Louisiana State University, has a long history of water quality problems, including algal blooms, fish kills, and high concentrations of fecal indicator bacteria. Periodic dredging of the lake is necessary to prevent its return to swampland. This study was undertaken to elucidate the roles of allochthonous versus autochthonous nutrients as causes of water quality problems in the lake, with the expectation that this information would help identify strategies for lake restoration. Photosynthetic rates and concentrations of inorganic nutrients and phytoplankton pigments were measured over a period of one year. More than 90% of the chlorophyll a (chl a) in the lake was accounted for by Chlorophyceae, Cyanophyceae, and Bacillariophyceae. Concentrations of chl a, which averaged 75 μg L−1, fluctuated weekly during dry weather by as much as a factor of four. Phytoplankton growth rates were about 30% higher 1–2 days after rain events than after periods of dry weather, the implication being that allochthonous nutrient loading has a significant effect on the dynamics of the phytoplankton community in the lake. Therefore, dredging of sediments will likely produce no long-term improvement in water quality. More than 100 storm drains currently discharge into the lake, and diversion of those drains may be the most cost-effective strategy for effecting a long-term improvement in water quality.

Influence of Sewered Versus Septic Systems on Watershed Exports of E. coli

Elevated bacteria concentrations have led to the impairment (e.g., closures of shellfisheries and recreational beaches) of coastal waters. Although many previous studies have suggested that wastewater inputs can lead to elevated fecal indicator bacteria (FIB) concentrations in surface waters, few studies have quantified wastewater-associated FIB exports at the watershed scale. The goal of this study was to estimate bacterial exports at the watershed scale based on wastewater management approach (septic vs. sewer). Six watersheds (three exclusively on septic and three exclusively served by a sewer system) were selected for water quality assessment and comparison. Streams were monitored approximately monthly from August 2011 to June 2012 during baseflow conditions. Additionally, three storms were monitored. Samples were collected in sterile 100-mL bottles and analyzed for Escherichia coli concentrations. Discharge from streams was measured and bacterial exports were estimated by multiplying discharge by E. coli concentration. The results revealed that (1) during baseflow conditions, septic watersheds contained elevated stream discharge and E. coli concentrations and exports as compared to sewer watersheds; (2) warmer months had elevated E. coli watershed exports compared to colder months in both septic and sewer watersheds; and (3) storms significantly increased watershed E. coli exports in both septic and sewer watersheds. Storms significantly increased watershed E. coli exports in both septic and sewered watersheds, but E. coli counts in sewered watersheds were considerably greater likely due to greater impervious surface coverage and or leaky sewer infrastructure. These findings in conjunction with previous studies suggest that septic systems may play a pivotal role in the delivery of FIB to receiving waters, particularly during baseflow conditions.

The Impact of Environmental Variables on Faecal Indicator Bacteria in the Betna River Basin, Bangladesh

Environmental variables influence Faecal Indicator Bacteria (FIB) in surface water. Understanding that influence is important, because presence of FIB, which are an indication of faecal contamination, means that harmful pathogens could be present that could also be influenced by environmental variables. Although some recent studies have focused on this topic, most of this work has been conducted in developed countries. Similar studies in developing countries and in a (sub)tropical climate are lacking. In this study we assess the influence of environmental variables on fluctuations in FIB concentrations of the Betna River in southwest Bangladesh that floods almost every year. Monthly water samples from five locations along Betna River were tested for FIB (E. coli and enterococci) in 2014–2015. A linear regression model was developed to assess the effect of the environmental variables on FIB concentrations. The study revealed increased FIB concentrations during wet weather conditions. Precipitation and water temperature were positively correlated with FIB concentrations. Water temperature was positively correlated, because the warm May to September period coincides with frequent precipitation. Precipitation increases manure release from land to surface water. The regression model explains nearly half of the variability in FIB concentrations (R2 of 0.46 for E. coli and 0.48 for enterococci). This study indicates that increased precipitation combined with higher water temperature, as is expected in this region with climate change, likely increases FIB concentrations. Waterborne pathogens are expected to respond similarly to these environmental changes, indicating that disease outbreaks could well become more frequent and severe.

Transport and attenuation of Salmonella enterica, fecal indicator bacteria and a poultry litter marker gene are correlated in soil columns

Millions of tons of fecal-contaminated poultry litter are applied to U.S. agricultural fields annually. Precipitation and irrigation facilitate transport of fecal-derived pathogens and fecal indicator bacteria (FIB) to groundwater. The goal of this study was to compare transport of pathogens, FIB, and a microbial source tracking marker gene for poultry litter (LA35) in a simulated soil-to-groundwater system. Nine laboratory soil columns containing four different soil types were used to evaluate microbial transport to groundwater via infiltration. Quantitative polymerase chain reaction was used to monitor Salmonella enterica Typhimurium, Escherichia coli, Enterococcus spp., Brevibacterium sp. LA35 and Bacteroidales leached from soil columns inoculated with poultry litter. S. enterica was correlated with LA35 poultry litter marker gene and FIB concentrations in column soils containing organic matter, but not in acid washed sands. In contrast, S. enterica was found to correlate with LA35 and FIB in the leachate from columns containing sand, but not with leachate from organic soil columns. The majority of recovered DNA was found in leachate of predominately sandy soil columns, and in the soil of loamy columns. At least 90% of the DNA retained in soils for each microbial target was found in the top 3cm of the column. These studies suggest that poultry litter associated pathogens and FIB are rapidly released from litter, but are influenced by complex attenuation mechanisms during infiltration, including soil type. This study advances our understanding of the potential for subsurface transport of poultry litter associated pathogens and FIB, and support the use of the LA35 marker gene for evaluating poultry litter impacts on groundwater.

Microbiological quality of water in a city with persistent and recurrent waterborne diseases under tropical sub-rural conditions: The case of Kikwit City, Democratic Republic of the Congo.

The availability of safe drinking water in sub-Saharan countries remains a major challenge because poor sanitation has been the cause of various outbreaks of waterborne disease due to the poor microbiological quality of water used for domestic purposes. The faecal indicator bacteria (FIB) used in the present study included Escherichia coli (E. coli) and Enterococcus (ENT). FIB and aerobic mesophilic bacteria (AMB) were quantified during July 2015 (dry season) and November 2015 (rainy season) in order to assess the quality of drinking water from wells (n=3; P1-P3), and two rivers, the River Lukemi (RLK, n=3) and River Luini (RLN, n=2) in the city of Kikwit, which is located in the province of Kwilu in the Democratic Republic of the Congo. Kikwit is well known for its outbreaks of persistent and recurrent waterborne diseases including Entamoeba, Shigella, typhoid fever, cholera, and Ebola Viral Hemorrhagic Fever. Consequently, E. coli, ENT, and AMB were quantified in water samples according to the standard international methods for water quality determination using the membrane filtration method. The FIB characterization was performed for human-specific Bacteroides by PCR using specific primers. The results obtained revealed high FIB concentrations in river samples collected during both seasons. For example, E. coli respectively reached 4.3×104 and 9.2×104 CFU 100mL-1 in the dry season and the wet season. ENT reached 5.3×103 CFU 100mL-1 during the dry season and 9.8×103 CFU 100mL-1 in the wet season. The pollution was significantly worse in the wet season compared to the dry season. Surprisingly, no faecal contamination was observed in well water samples collected in the dry season while E. coli and ENT were detected in all wells in the wet season with values of 6, 7, and 11CFUmL-1 for E. coli in wells P1-P3, respectively and 3, 5, 9 CFU mL-1for ENT in the same wells. Interestingly, the PCR assays for human-specific Bacteroides HF183/HF134 indicated that 97-100% captured in all analyses of isolated FIB were of human origin. The results indicate that contamination of E. coli, ENT, and AMB in the studied water resources increases during the wet season. This study improves understanding of the microbiological pollution of rivers and wells under tropical conditions and will guide future municipal/local government decisions on improving water quality in this region which is characterised by persistent and recurrent waterborne diseases. Although the epidemiology can be geographically localised, the effects of cross border transmission can be global. Therefore, the research results presented in this article form recommendations to municipalities/local authorities and the approach and procedures can be carried out in a similar environment.

The compartment bag test (CBT) for enumerating fecal indicator bacteria: Basis for design and interpretation of results

For the past several years, the compartment bag test (CBT) has been employed in water quality monitoring and public health protection around the world. To date, however, the statistical basis for the design and recommended procedures for enumerating fecal indicator bacteria (FIB) concentrations from CBT results have not been formally documented. Here, we provide that documentation following protocols for communicating the evolution of similar water quality testing procedures. We begin with an overview of the statistical theory behind the CBT, followed by a description of how that theory was applied to determine an optimal CBT design. We then provide recommendations for interpreting CBT results, including procedures for estimating quantiles of the FIB concentration probability distribution, and the confidence of compliance with recognized water quality guidelines. We synthesize these values in custom user-oriented ‘look-up’ tables similar to those developed for other FIB water quality testing methods. Modified versions of our tables are currently distributed commercially as part of the CBT testing kit.

Microbial source tracking in shellfish harvesting waters in the Gulf of Nicoya, Costa Rica

Current microbial water quality monitoring is generally limited to culture-based measurements of fecal indicator bacteria (FIB). Given the many possible sources of fecal pollution within a watershed and extra-intestinal FIB reservoirs, it is important to determine source(s) of fecal pollution as a means to improve water quality and protect public health. The principal objective of this investigation was to characterize the microbial water quality of shellfish harvesting areas in the Gulf of Nicoya, Costa Rica during 2015. In order to achieve this objective, the specificity and sensitivity of 11 existing microbial source tracking (MST) PCR assays, associated with cows (BacCow), dogs (BacCan, DogBac), domestic wastewater (PMMoV), general avian (GFD), gulls (Gull2), horses (HorseBac, HoF), humans (HF183, HPyV), and pigs (PF), were evaluated using domestic wastewater and animal fecal samples collected from the region. The sensitivity of animal-associated assays ranged from 13 to 100%, while assay specificity ranged from 38 to 100%. The specificity of pepper mild mottle virus (PMMoV) and human polyomavirus (HPyV) was 100% for domestic wastewater, as compared to 94% specificity of the HF183 Bacteroidales marker. PMMoV was identified as a useful domestic wastewater-associated marker, with concentrations as high as 1.1 × 105 copies/ml and 100% sensitivity and specificity. Monthly surface water samples collected from four shellfish harvesting areas were analyzed using culture-based methods for Escherichia coli as well as molecular methods for FIB and a suite of MST markers, which were selected for their specificity in the region. While culturable E. coli results suggested possible fecal pollution during the monitoring period, the absence of human/domestic wastewater-associated markers and low FIB concentrations determined using molecular methods indicated sufficient microbial water quality for shellfish harvesting. This is the first study to our knowledge to test the performance of MST markers in Costa Rica as well as in Central America. Given the lack of wastewater treatment and the presence of secondary sources of FIB, this study highlights the importance of an MST toolbox approach to characterize water quality in tropical regions. Furthermore, it confirms and extends the geographic range of PMMoV as an effective tool for monitoring domestic wastewater pollution.

Spatial and temporal microbial pollution patterns in a tropical estuary during high and low river flow conditions

Spatial and temporal patterns of coastal microbial pollution are not well documented. Our study examined these patterns through measurements of fecal indicator bacteria (FIB), nutrients, and physiochemical parameters in Hilo Bay, Hawai'i, during high and low river flow. >40% of samples tested positive for the human-associated Bacteroides marker, with highest percentages near rivers. Other FIB were also higher near rivers, but only Clostridium perfringens concentrations were related to discharge. During storms, FIB concentrations were three times to an order of magnitude higher, and increased with decreasing salinity and water temperature, and increasing turbidity. These relationships and high spatial resolution data for these parameters were used to create Enterococcus spp. and C. perfringens maps that predicted exceedances with 64% and 95% accuracy, respectively. Mapping microbial pollution patterns and predicting exceedances is a valuable tool that can improve water quality monitoring and aid in visualizing FIB hotspots for management actions.

Escherichia coli Reduction by Bivalves in an Impaired River Impacted by Agricultural Land Use.

Fecal indicator bacteria (FIB) are leading causes of impaired surface waters. Innovative and environmentally appropriate best management practices are needed to reduce FIB concentrations and associated risk. This study examines the ability of the native freshwater mussel Anodonta californiensis and an invasive freshwater clam Corbicula fluminea to reduce concentrations of the FIB Escherichia coli in natural waters. Laboratory batch experiments were used to show bivalve species-specific E. coli removal capabilities and to develop a relationship between bivalve size and clearance rates. A field survey within an impaired coastal river containing both species of bivalves in an agricultural- and grazing-dominated area of the central coast of California showed a significant inverse correlation between E. coli concentration and bivalve density. An in situ field spiking and sampling study showed filtration by freshwater bivalves resulting in 1-1.5 log10 reduction of E. coli over 24 h, and calculated clearance rates ranged from 1.2 to 7.4 L hr-1 bivalve-1. Results of this study show the importance of freshwater bivalves for improving water quality through the removal of E. coli. While both native and invasive bivalves can reduce E. coli levels, the use of native bivalves through integration into best management practices is recommended as a way to improve water quality and protect and encourage re-establishment of native bivalve species that are in decline.

Watershed Assessment with Beach Microbial Source Tracking and Outcomes of Resulting Gull Management.

Total maximum daily load (TMDL) implementation at a southern California beach involved ultraviolet treatment of watershed drainage that provided >97% reduction in fecal indicator bacteria (FIB) concentrations. However, this pollutant control measure did not provide sufficient improvement of beach water quality, prompting further assessment. Investigation included microbial source tracking (MST) for human, gull, and canine fecal sources, monitoring of enterococci and fecal coliform, and measurement of chemical and physical water quality parameters for samples collected from watershed, groundwater, and beach sites, including a beach scour pond and tidal creek. FIB variability remained poorly modeled in regression analysis. However, MST revealed correlations between FIB and gull source tracking markers, leading to recommendations to manage gulls as a pollutant source. Beach conditions were followed for three years after implementation of a best management practice (BMP) to abate gulls using a falconry program for the beach and an upland landfill. The gull abatement BMP was associated with improved beach water quality, and this appears to be the first report of falconry in the context of TMDL implementation. Overall, MST data enabled management action despite an inability to fully model FIB dynamics in the coupled watershed-beach system.

The impact of hospital and urban wastewaters on the bacteriological contamination of the water resources in Kinshasa, Democratic Republic of Congo

ABSTRACTAlthough the United Nations General Assembly recognized in 2010 the right to safe and clean drinking water and sanitation as a human right that is essential to the full enjoyment of life and all other human rights, the contamination of water supplies with faecal pathogens is still a major and unsolved problem in many parts of the world. In this study, faecal indicator bacteria (FIB), including Escherichia coli (E. coli) and Enterococcus (ENT), were quantified over the period of June/July 2014 and June/July 2015 to assess the quality of hospital effluents (n = 3: H1, H2 and H3) and of rivers receiving wastewaters from the city of Kinshasa, Democratic Republic of Congo. The water and sediment samples from the river-receiving systems were collected in, upstream and downstream of the hospital outlet pipe (HOP) discharge. The analysis of E. coli and ENT in water and sediment suspension was performed using the cultural membrane filter method. The FIB characterization was performed for general E. coli, Enterococcus faecalis(E. faecalis) and human-specific Bacteroides by PCR using specific primers. The results revealed very high FIB concentration in the hospital effluent waters, with E. coli reaching the values of 4.2 × 105, 16.1 × 105 and 5.9 × 105 CFU 100 mL−1, for the hospital effluents from H1, H2, and H3, respectively; and Enterococcus reaching the values of 2.3 × 104, 10.9 × 104 and 4.1 × 104 CFU 100 mL−1, respectively. Interestingly, the FIB levels in the water and sediment samples from river-receiving systems are spatially and temporally highly variable and present in some samples with higher values than the hospital effluents. The PCR assays for human-specific Bacteroides HF183/HF134 further indicate that more than 98% of bacteria were from human origin. The results of this research therefore confirm the hypothesis of our previous studies, indicating that in developing countries (e.g., Democratic Republic of Congo and South India), the hospital effluent waters can be a significant source of the deterioration of the bacteriological quality for urban rivers. The approach used in this investigation can be further used to decipher the pollution of water resources by human faecal contamination. The results of this research will help to better understand the microbiological pollution problems in river-receiving systems and will guide municipality decisions on improving the urban water quality.

Fecal contamination and Microcystis in drinking-water sources of rural Cambodia using PCR and culture-based methods

Rural communities within low-income countries frequently rely on a range of drinking-water sources, and each water source varies in its potential for biological contamination. The extent and source of biological contamination in primary drinking sources within Kien Svay, Kandal, Cambodia, were determined by fecal indicator bacteria (FIB) measurements, 16S rDNA genetic markers for human and bovine fecal Bacteroides, presence of the bloom-forming Microcystis species, and the microcystin toxin mcyD gene marker. Thirteen wells, 11 rain barrels, 10 surface-water sites, and five sediment samples were examined during the dry and wet seasons. Surface water was commonly contaminated with FIB, with up to 1.02 × 105Enterococcus sp., 6.13 × 104E. coli, and 2.91 × 104 total coliforms per 100 mL of water. Human and bovine Bacteroides were detected in 100 and 90% of the surface water samples, respectively. Concentrations of FIB in rain-barrels varied by site, however 91% contained human Bacteroides. Microcystis cells were found in 90% of surface water sites, with many also containing microcystin gene mcyD, representing the first report of microcystin-producing cyanobacteria in surface waters of Cambodia. The study results show that many potential drinking-water sources in Cambodia contain harmful bacterial and algal contaminants, and care should be taken when selecting and monitoring water options.

Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed

Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial source tracking (MST) markers, developed to determine potential sources of fecal contamination, can also be resuspended from bed sediments. The primary objective of this study was to predict occurrence of waterborne pathogens in water and streambed sediments using a simple statistical model that includes traditionally measured FIB, environmental parameters and source allocation, using MST markers as predictor variables. Synoptic sampling events were conducted during baseflow conditions downstream from agricultural (AG), forested (FORS), and wastewater pollution control plant (WPCP) land uses. Concentrations of FIB and MST markers were measured in water and sediments, along with occurrences of the enteric pathogens Campylobacter, Listeria and Salmonella, and the virulence gene that carries Shiga toxin, stx2. Pathogens were detected in water more often than in underlying sediments. Shiga toxin was significantly related to land use, with concentrations of the ruminant marker selected as an independent variable that could correctly classify 76% and 64% of observed Shiga toxin occurrences in water and sediment, respectively. FIB concentrations and water quality parameters were also selected as independent variables that correctly classified Shiga toxin occurrences in water and sediment (54%–87%), and Salmonella occurrences in water (96%). Relationships between pathogens and indicator variables were generally inconsistent and no single indicator adequately described occurrence of all pathogens. Because of inconsistent relationships between individual pathogens and FIB/MST markers, incorporating a combination of FIB, water quality measurements, and MST markers may be the best way to assess microbial water quality in mixed land use systems.

Evaluation of Enterococcus concentrations at beaches in Door County, WI (Lake Michigan, USA) by qPCR and defined substrate culture analysis

The US EPA (Environmental Protection Agency) has approved the use of a quantitative polymerase chain reaction (qPCR) method for surface water regulatory decisions. This method would allow for results in as little as 4 h, rather than the 24 + hours for culture-based tests. The US EPA qPCR Method 1611 is enterococci-based, whereas the Great Lakes states adopted Escherichia coli as the indicator organism of choice. The change in indicator organism, method, and the heterogeneous nature of Great Lake beaches present challenges to implementation of rapid methods. The objective of this study was to determine the feasibility of using qPCR for beach water monitoring and management in rural, northern Lake Michigan communities. Concentrations of fecal indicator bacteria (FIB) were different in each method and varied greatly by beach. When concentrations of enterococci by TaqMan Enterococcus qPCR (EntTaq) were compared with E. coli using Colilert, positive correlations were detected only at Baileys Harbor, Fish Creek and Sunset Park Beaches (r2 = 0.26, r2 = 0.22 and r2 = 0.36, respectively). When enterococci concentrations determined by EntTaq were used for management decisions, the EntTaq method would have resulted in 47 false positive and 27 false negative results when compared to culture-based decisions. When management decisions based on E. coli concentrations were obtained from Colilert, EntTaq would have resulted in 35 false positive and 49 false negative results. This study suggests that, although minimally trained individuals can obtain appropriate results using qPCR methods, each beach community should carefully evaluate the impact of adopting rapid methods for beach management decisions.

Fecal Indicator Bacteria Entrainment from Streambed to Water Column: Transport by Unsteady Flow over a Sand Bed

Storms cause a substantial increase in the fecal indicator bacteria (FIB) concentrations in stream water as a result of FIB-laden runoff and the release of FIB from stream sediments. Previous work has emphasized the association between FIB and bed sediments finer than sand. The objectives of this work were to elucidate the effect of various velocities on the entrainment of bed-dwelling coliforms in sand-bed streams and to refine methodologies for quantifying sandy streambeds as sources of FIB. Pump-induced hydrographs were created using a stainless steel nonrecirculating flume. Experiments consisted of simulating four storm hydrographs and collecting water samples upstream and downstream of a sand bed at selected intervals. Bed sediment samples were collected before and after each event. The highest concentrations of total coliform and suspended sediments generally occurred in the downstream samples during the rising limb of the hydrographs as a result of entrainment of coliforms and sand from the bed to the water column. There was a first flush effect in the system, as the upper layer of sand was influenced by a rapidly increasing velocity at ∼0.2 m s. Coliforms downstream of the sand bed increased rapidly as velocity exceeded this threshold but then declined even as velocity and discharge continued to increase. This likely reflects the depletion of coliforms as the more densely populated sediment layer was flushed out. There is evidence that streams with sand beds harbor enough FIB that development of total maximum daily loads (TMDLs) should include consideration of them as a source.

Contamination with bacterial zoonotic pathogen genes in U.S. streams influenced by varying types of animal agriculture

Animal waste, stream water, and streambed sediment from 19 small (<32km2) watersheds in 12U.S. states having either no major animal agriculture (control, n=4), or predominantly beef (n=4), dairy (n=3), swine (n=5), or poultry (n=3) were tested for: 1) cholesterol, coprostanol, estrone, and fecal indicator bacteria (FIB) concentrations, and 2) shiga-toxin producing and enterotoxigenic Escherichia coli, Salmonella, Campylobacter, and pathogenic and vancomycin-resistant enterococci by polymerase chain reaction (PCR) on enrichments, and/or direct quantitative PCR. Pathogen genes were most frequently detected in dairy wastes, followed by beef, swine and poultry wastes in that order; there was only one detection of an animal-source-specific pathogen gene (stx1) in any water or sediment sample in any control watershed. Post-rainfall pathogen gene numbers in stream water were significantly correlated with FIB, cholesterol and coprostanol concentrations, and were most highly correlated in dairy watershed samples collected from 3 different states. Although collected across multiple states and ecoregions, animal-waste gene profiles were distinctive via discriminant analysis. Stream water gene profiles could also be discriminated by the watershed animal type. Although pathogen genes were not abundant in stream water or streambed samples, PCR on enrichments indicated that many genes were from viable organisms, including several (shiga-toxin producing or enterotoxigenic E. coli, Salmonella, vancomycin-resistant enterococci) that could potentially affect either human or animal health. Pathogen gene numbers and types in stream water samples were influenced most by animal type, by local factors such as whether animals had stream access, and by the amount of local rainfall, and not by studied watershed soil or physical characteristics. Our results indicated that stream water in small agricultural U.S. watersheds was susceptible to pathogen gene inputs under typical agricultural practices and environmental conditions. Pathogen gene profiles may offer the potential to address both source of, and risks associated with, fecal pollution.

Restructuring of the Aquatic Bacterial Community by Hydric Dynamics Associated with Superstorm Sandy.

Bacterial community composition and longitudinal fluctuations were monitored in a riverine system during and after Superstorm Sandy to better characterize inter- and intracommunity responses associated with the disturbance associated with a 100-year storm event. High-throughput sequencing of the 16S rRNA gene was used to assess microbial community structure within water samples from Muddy Creek Run, a second-order stream in Huntingdon, PA, at 12 different time points during the storm event (29 October to 3 November 2012) and under seasonally matched baseline conditions. High-throughput sequencing of the 16S rRNA gene was used to track changes in bacterial community structure and divergence during and after Superstorm Sandy. Bacterial community dynamics were correlated to measured physicochemical parameters and fecal indicator bacteria (FIB) concentrations. Bioinformatics analyses of 2.1 million 16S rRNA gene sequences revealed a significant increase in bacterial diversity in samples taken during peak discharge of the storm. Beta-diversity analyses revealed longitudinal shifts in the bacterial community structure. Successional changes were observed, in which Betaproteobacteria and Gammaproteobacteria decreased in 16S rRNA gene relative abundance, while the relative abundance of members of the Firmicutes increased. Furthermore, 16S rRNA gene sequences matching pathogenic bacteria, including strains of Legionella, Campylobacter, Arcobacter, and Helicobacter, as well as bacteria of fecal origin (e.g., Bacteroides), exhibited an increase in abundance after peak discharge of the storm. This study revealed a significant restructuring of in-stream bacterial community structure associated with hydric dynamics of a storm event. In order to better understand the microbial risks associated with freshwater environments during a storm event, a more comprehensive understanding of the variations in aquatic bacterial diversity is warranted. This study investigated the bacterial communities during and after Superstorm Sandy to provide fine time point resolution of dynamic changes in bacterial composition. This study adds to the current literature by revealing the variation in bacterial community structure during the course of a storm. This study employed high-throughput DNA sequencing, which generated a deep analysis of inter- and intracommunity responses during a significant storm event. This study has highlighted the utility of applying high-throughput sequencing for water quality monitoring purposes, as this approach enabled a more comprehensive investigation of the bacterial community structure. Altogether, these data suggest a drastic restructuring of the stream bacterial community during a storm event and highlight the potential of high-throughput sequencing approaches for assessing the microbiological quality of our environment.

Decay rates of faecal indicator bacteria from sewage and ovine faeces in brackish and freshwater microcosms with contrasting suspended particulate matter concentrations

To safeguard human health, legislative measures require the monitoring of faecal indicator bacteria (FIB) concentrations in recreational and shellfish waters. Consequently, numerous studies have focussed on FIB survival in the water column and more recently in estuarine sediments. However, there is a paucity of information regarding the influence of contrasting suspended particulate matter (SPM) concentrations on the survival of FIB in the water column of estuaries. Here, microcosms containing freshwater or brackish water with low, high and extreme SPM concentrations were inoculated with sewage and ovine faeces and the decay rate of Escherichia coli, coliforms and enterococci were determined by enumeration over five consecutive days. E. coli derived from ovine faeces proliferated and persisted at high levels in both freshwater and brackish microcosms (no decay), whereas ovine enterococci demonstrated a net decay over the duration of the experiment. Furthermore, SPM concentration had a significant effect on the decay rates of both E. coli and enterococci from ovine faeces in brackish microcosms, but decay rate was greater at low SPM concentrations for E. coli, whereas the opposite was observed for enterococci, whose decay rates increased as SPM concentration increased. E. coli, enterococci and coliforms derived from wastewater demonstrated a net decay in both freshwater and brackish microcosms, with contrasting effects of SPM concentration on decay rate. In addition, some FIB groups demonstrated contrasting responses (decay or proliferation) in the first 24h following inoculation into freshwater versus brackish microcosms. Overall, SPM concentrations influenced the proliferation and decay rates of FIB in brackish waters, but had minimal influence in freshwater. These results demonstrate that the survival rates of FIB in aquatic environments are system specific, species and source dependent, and influenced by SPM concentration. This study has important implications for catchment-based risk assessments and source apportionment of FIB pollution in aquatic environments.

Fecal Indicator Bacteria Dynamics in a Surface Flow Constructed Wetland in Southwestern Illinois, USA

Constructed wetlands have been used to treat wastewater because of their efficiency at removing fecal indicator bacteria (FIB), their low cost, and their ease of maintenance. This study investigates the ability of a surface-flow wetland (0.03 ha in size), constructed in the rural village of Elsah, Illinois, to treat FIB pollution. The objectives of this study were to: 1) compare mean FIB concentrations (specifically thermotolerant coliforms [TTC] and enterococci [ENT]) in the wetland during low versus high precipitation conditions; 2) compare mean FIB levels among different sampling locations along the wetland’s treatment gradient; and 3) determine whether FIB and other environmental variables were significantly correlated. Both TTC and ENT levels increased during storm events, likely due to increased mobilization of sediment. Both TTC and ENT were significantly lower in zones located further from the inflow point. The strongest correlation was observed between TTC and ENT, and both parameters were strongly correlated with precipitation.