Significant Factors for Modelling Survival of Escherichia coli in Lake Sediments.

Currently available numerical models that describe the fecal contamination of aquatic environments using Escherichia coli as an indicator bacterium did not consider its survival in sediments. We conducted a series of comparative experiments to reveal the independent and interactive effects of sediment factors, including temperature, pH, water-extractable total dissolved solids (TDSs), coexisting microbes, and sampling sites, in lake environments on E. coli survival. In experiments, E. coli survival was observed by controlling any two factors at a time. Consequently, the decrease in pH and presence of coexisting microbes enhanced E. coli die-off, whereas the addition of water-extractable TDSs promoted its growth. To select factors to be considered for modelling E. coli survival in sediments, the independent effects of each factor and the interaction effect of the two factors were statistically compared based on their effect sizes (η2). As a result, pH (η2 = 59.5-89.0%) affected E. coli survival most significantly, followed by coexisting microbes (1.7-48.4%). Among the interactions affecting E. coli survival, including pH or coexisting microbes-which had larger independent effects-relatively larger statistically significant interactions were observed between pH and coexisting microbes (31.1%), coexisting microbes and water-extractable TDSs (85.4%), and coexisting microbes and temperature (26.4%).

Assessing the utility of shellfish sanitation monitoring data for long-term estuarine water quality analysis.

Regular testing of coastal waters for fecal coliform bacteria by shellfish sanitation programs could provide data to fill large gaps in existing coastal water quality monitoring, but research is needed to understand the opportunities and limitations of using these data for inference of long-term trends. In this study, we analyzed spatiotemporal trends from multidecadal fecal coliform concentration observations collected by a shellfish sanitation program, and assessed the feasibility of using these monitoring data to infer long-term water quality dynamics. We evaluated trends in fecal coliform concentrations for a 20-year period (1999-2021) using data collected from spatially fixed sampling sites (n = 466) in North Carolina (USA). Findings indicated that shellfish sanitation data can be used for long-term water quality inference under relatively stationary management conditions, and that salinity trends can be used to investigate management-driven bias in fecal coliform observations collected in a particular area.

Two-year Monitoring of Microbiological Water Quality in Small Water Supply Systems: Implications for Microbial Risk Management.

Small water supply systems (SWSSs) are often more vulnerable to waterborne disease outbreaks. In Japan, many SWSSs operate without regulation under the Waterworks Law, yet there is limited investigation into microbial contamination and the associated health risks. In this study, the microbiological water quality of four SWSSs that utilize mountain streams as water sources and do not install water treatment facilities were monitored for over 2 years. In investigated SWSSs, the mean heterotrophic plate counts were below 350 CFU/mL, and the total bacterial loads (16S rDNA concentration) ranged from 4.71 to 5.35 log10 copies/mL. The results also showed the consistent presence of fecal indicator bacteria (FIB), i.e., Escherichia coli and Clostridium perfringens, suggesting the potential of fecal pollution. E. coli was then utilized as an indicator to assess the health risk posed by E. coli O157:H7 and Campylobacter jejuni. The results indicated that the estimated mean annual risk of infection and disability-adjusted life years (DALYs) exceeded acceptable levels in all SWSSs for the two reference pathogens. To ensure microbial water safety, implementing appropriate water treatment facilities with an estimated mean required reduction of 5-6 log10 was necessary. This study highlighted the potential microbial contamination and health risk level in SWSSs that utilize mountain streams as water sources, even though the water sources were almost not affected by human activities. Furthermore, this study would also be helpful in supporting risk-based water management to ensure a safe water supply in SWSSs.

Influence of combined abiotic/biotic factors on decay of P. aeruginosa and E. coli in Rhine River water.

Understanding the dynamic change in abundance of both fecal and opportunistic waterborne pathogens in urban surface water under different abiotic and biotic factors helps the prediction of microbiological water quality and protection of public health during recreational activities, such as swimming. However, a comprehensive understanding of the interaction among various factors on pathogen behavior in surface water is missing. In this study, the effect of salinity, light, and temperature and the presence of indigenous microbiota, on the decay/persistence of Escherichia coli and Pseudomonas aeruginosa in Rhine River water were tested during 7 days of incubation with varying salinity (0.4, 5.4, 9.4, and 15.4 ppt), with light under a light/dark regime (light/dark) and without light (dark), temperature (3, 12, and 20 °C), and presence/absence of indigenous microbiota. The results demonstrated that light, indigenous microbiota, and temperature significantly impacted the decay of E. coli. Moreover, a significant (p<0.01) four-factor interactive impact of these four environmental conditions on E. coli decay was observed. However, for P. aeruginosa, temperature and indigenous microbiota were two determinate factors on the decay or growth. A significant three-factor interactive impact between indigenous microbiota, temperature, and salinity (p<0.01); indigenous microbiota, light, and temperature (p<0.01); and light, temperature, and salinity (p<0.05) on the decay of P. aeruginosa was found. Due to these interactive effects, caution should be taken when predicting decay/persistence of E. coli and P. aeruginosa in surface water based on a single environmental condition. In addition, the different response of E. coli and P. aeruginosa to the environmental conditions highlights that E. coli monitoring alone underestimates health risks of surface water by non-fecal opportunistic pathogens, such as P. aeruginosa. KEY POINTS: Abiotic and biotic factors interactively affect decay of E. coli and P. aeruginosa E.coli and P.aeruginosa behave significantly different under the given conditions Only E. coli as an indicator underestimates the microbiological water quality.

Village Settlements in Mountainous Tropical Areas, Hotspots of Fecal Contamination as Evidenced by Escherichia coli and Stanol Concentrations in Stormwater Pulses.

Fecal bacteria in surface water may indicate threats to human health. Our hypothesis is that village settlements in tropical rural areas are major hotspots of fecal contamination because of the number of domestic animals usually roaming in the alleys and the lack of fecal matter treatment before entering the river network. By jointly monitoring the dynamics of Escherichia coli and of seven stanol compounds during four flood events (July-August 2016) at the outlet of a ditch draining sewage and surface runoff out of a village of Northern Lao PDR, our objectives were (1) to assess the range of E. coli concentration in the surface runoff washing off from a village settlement and (2) to identify the major contributory sources of fecal contamination using stanol compounds during flood events. E. coli pulses ranged from 4.7 × 104 to 3.2 × 106 most probable number (MPN) 100 mL-1, with particle-attached E. coli ranging from 83 to 100%. Major contributory feces sources were chickens and humans (about 66 and 29%, respectively), with the highest percentage switching from the human pole to the chicken pole during flood events. Concentrations indicate a severe fecal contamination of surface water during flood events and suggest that villages may be considered as major hotspots of fecal contamination pulses into the river network and thus as point sources in hydrological models.

Assessing the defecation practices of unsheltered individuals and their contributions to microbial water quality in an arid, urban watershed.

Outdoor defecation by people experiencing homelessness is frequently perceived as a potentially large source of human fecal pollution and a significant source of health risk in urban waterbodies with recreational contact. The goal of this study was to count the number of people experiencing homelessness and quantifies their sanitation habits in an urban river corridor setting, then use this information for estimating human fecal pollutant loading on a watershed scale. Two types of census counts were conducted including periodic point-in-time counts over six years and weekly counts of encampments. While the population census varied from count-to-count, the range of population estimates in the river corridor varied from 109 to 349 individuals during the six-year span, which mirrored the weekly counts of encampments. A face-to-face survey of people experiencing homelessness assessed the sanitation habits of the unsheltered population (N = 63), including outdoor defecation frequency and containment practices. Overall, 95 % of survey respondents reported defecating outdoors; 36 % practiced outdoor defecation between 4 and 7 days/week and 27 % practiced outdoor defecation <1 day/week. Of those that did practice outdoor defecation, 75 % contained their feces in a bucket or bag, thereby limiting fecal material contributions to the river; 6.7 % reported defecating on low ground near the river that could wash off when flood waters rise during a storm event. Only a single survey respondent reported defecating directly into the river. Based on literature values for average HF183 output for an adult human, and the average rainfall in the urban watershed, the total watershed contribution of HF183 averaged 1.2 × 1010 gene copies per storm event (95 % CI: 0.9 × 1010-1.6 × 1010) along the 41 km stretch of river in this study. This human fecal loading estimate is at least two orders of magnitude less than cumulative HF183 loading from all human sources measured at the bottom of the watershed.

Vertical stratification and seasonality of fecal indicator bacteria in New York City playground sandboxes.

Sandboxes in public play spaces afford a crucial opportunity for urban children to engage in naturalistic play that fosters development of cognitive, social, and motor skills. As open pits, sandboxes in New York City public playgrounds are potentially exposed to fecal inputs from various sources, including wild and domestic animals. A longitudinal study of thirteen sandboxes located in public playgrounds on the east side of Manhattan reveals ubiquity of the fecal indicator bacteria enterococci and Escherichia coli through all seasons. The highest concentrations of bacteria occur in surface sand (n = 42; mean enterococci 230 MPN/g and E. coli 182 MPN/g dry weight), with significantly lower levels at depths below the surface (n = 35; mean enterococci 21 MPN/g and E. coli 12 MPN/g dry weight), a stratification consistent with fecal loading at the surface. Generalized linear mixed models indicate that sand depth (surface vs. underlayers) is the most influential variable affecting bacterial levels (P <0.001 for both enterococci and E. coli), followed by sampling season (P <0.001 for both). Bacterial concentrations do not vary significantly as a function of playground location or ZIP code within the study area. Children's exposure while playing in sandboxes likely reaches 105 enterococci and 104E. coli in a typical play period. Microbial source tracking to identify fecal hosts reveals dog, bird, and human biomarkers in low concentrations. Open sandbox microcosms installed at ground level in the urban environment of Manhattan are fouled by enterococci and E. coli within two weeks, while adjacent closed microcosms exhibit no fecal contamination over a 33-day sampling period. Collectively, our results indicate that increasing the frequency of sand refills and covering sandboxes during times of disuse would be straightforward management strategies to mitigate fecal contamination in playground sandboxes.

Short-term forecasting of fecal coliforms in shellfish growing waters.

This study sought to develop models for predicting near-term (1-3 day) fecal contamination events in coastal shellfish growing waters. Using Random Forest regression, we (1) developed fecal coliform (FC) concentration models for shellfish growing areas using watershed characteristics and antecedent hydrologic and meteorologic observations as predictors, (2) tested the change in model performance associated when forecasted, as opposed to measured, rainfall variables were used as predictors, and (3) evaluated model predictor importance in relation to shellfish sanitation management criteria. Models were trained to 10 years of coastal FC measurements (n = 1285) for 5 major shellfish management areas along the Florida (USA) coast. Model performance varied between the 5 management areas with R2 ranging from 0.36 to 0.72. Antecedent precipitation variables were among the most important predictors in the day-of forecast models in all management areas. When forecasted rainfall was included in the models, wind components became increasingly important.

Fecal Impairment Framework, A New Conceptual Framework for Assessing Fecal Contamination in Recreational Waters.

Fecal pollution of surface water is a pervasive problem that negatively affects waterbodies concerning both public health and ecological functions. Current assessment methods monitor fecal indicator bacteria (FIB) to identify pollution sources using culture-based quantification and microbial source tracking (MST). These types of information assist stakeholders in identifying likely sources of fecal pollution, prioritizing them for remediation, and choosing appropriate best management practices. While both culture-based quantification and MST are useful, they yield different kinds of information, potentially increasing uncertainty in prioritizing sources for management. This study presents a conceptual framework that takes separate human health risk estimates based on measured MST and E. coli concentrations as inputs and produces an estimate of the overall fecal impairment risk as its output. The proposed framework is intended to serve as a supplemental screening tool for existing monitoring programs to aid in identifying and prioritizing sites for remediation. In this study, we evaluated the framework by applying it to two primarily agricultural watersheds and several freshwater recreational beaches using existing routine monitoring data. Based on a combination of E. coli and MST results, the proposed fecal impairment framework identified four sites in the watersheds as candidates for remediation and identified temporal trends in the beach application. As these case studies demonstrate, the proposed fecal impairment framework is an easy-to-use and cost-effective supplemental screening tool that provides actionable information to managers using existing routine monitoring data, without requiring specialized expertize.

Integrating microbial source tracking with quantitative microbial risk assessment to evaluate site specific risk based thresholds at two South Florida beaches.

Quantitative microbial risk assessment (QMRA) can be used to evaluate health risks associated with recreational beach use. This study developed a site-specific risk assessment using a novel approach that combined quantitative PCR-based measurement of microbial source tracking (MST) genetic markers (human, dog, and gull fecal bacteria) with a QMRA analysis of potential pathogen risk. Water samples (n = 24) from two recreational beaches were collected and analyzed for MST markers as part of a broader Beach Exposure And Child Health Study that examined child behavior interactions with the beach environment. We report here the measurements of fecal bacteria MST markers in the environmental DNA extracts of those samples and a QMRA analysis of potential health risks utilizing the results from the MST measurements in the water samples. Human-specific Bacteroides was enumerated by the HF183 Taqman qPCR assay, gull-specific Catellicoccus was enumerated by the Gull2 qPCR assay, and dog-specific Bacteroides was enumerated by the DogBact qPCR assay. Derived reference pathogen doses, calculated from the MST marker concentrations detected in recreational waters, were used to estimate the risk of gastrointestinal illness for both children and adults. Dose-response equations were used to estimate the probability of the risk of infection (Pinf) per a swimming exposure event. Based on the QMRA simulations presented in this study, the GI risk from swimming or playing in water containing a mixture of human and non-human fecal sources appear to be primarily driven by the human fecal source. However, the estimated median GI health risk for both beaches never exceeded the U.S. EPA risk threshold of 32 illnesses per 1,000 recreation events. Our research suggests that utilizing QMRA together with MST can further extend our understanding of potential recreational bather risk by identifying the source contributing the greatest risk in a particular location, therefore informing beach management responses and decision-making.

Land use and rainfall influences on bacterial levels and sources in stormwater ponds.

Urban stormwater runoff is a known source of microbial contamination of stormwater ponds. However, less is known about the influences of land use and rainfall on microbial quality over time in these receiving waters. In this study, two fecal indicator bacteria (FIB), namely Escherichia coli and thermotolerant coliforms, were monitored in three stormwater ponds in Calgary, Alberta, Canada. The stormwater ponds were selected due to their potential as water sources for beneficial uses such as irrigation, which requires lower water quality than drinking water, thereby alleviating the pressure on the city's potable water demands. The selected stormwater ponds vary in size and shape, contribution catchment size, and percentages of several primary land use types. Microbial source tracking for human, dog, seagull, Canada goose, ruminant, and muskrat was also conducted to determine sources of bacterial contamination in the stormwater ponds. Sampling was conducted near the pond surface and adjacent to the shoreline, specifically near the outfalls that discharge stormwater runoff into the ponds and the inlets that convey water out of the ponds. Overall, the FIB concentrations in the vicinity of pond outfalls were significantly or relatively higher than those near pond inlets. The contamination in the McCall Lake and the Country Hills stormwater ponds showed higher amounts of human markers (40 to 60%) compared to the Inverness stormwater pond (< 20%), which coincided with their higher FIB concentration medians. The results revealed that stormwater drained from catchments with a higher percentage of commercial land use was more contaminated than those with primary residential land use, while the impacts of residential development on the FIB levels in the Inverness stormwater pond were not obvious. Furthermore, FIB concentrations in the ponds increased in response to both rain events and inter-event dry periods, with human-specific markers being predominant despite the high levels of animal markers during inter-event dry periods. Human-origin sources might be among the main microbial loading contributors in the pond catchments in general. All these findings can inform the development or improvement of measures for mitigating microbial pollution, strategies for reusing stormwater, and maintenance programs.

Examining the influence of COVID-19 lockdowns on coastal water quality: A study on fecal bacteria levels in Moroccan seawaters.

Fecal bacteria in bathing seawater pose a substantial public health risk and require rigorous monitoring. The unexpected beach closures during the COVID-19 lockdowns have afforded unique opportunities to evaluate the impact of human activities on bathing water quality (BWQ). This study examined the temporal changes in fecal coliforms (FC) and streptococci (FS) within bathing seawater across a popular coastal region in Morocco during two lockdown periods (2020 L and 2021 L), comparing these data with observations from pre-lockdown years (2018, 2019) and post-lockdown periods (2020, 2021, 2022). Our findings illuminate the influential role the hiatus periods played in enhancing bathing water quality, attaining an "excellent" status with marked reductions in fecal coliform and streptococci levels. Consequently, the FC/FS analysis exposed a clear preponderance of humans as the primary sources of fecal contamination, a trend that aligns with the burgeoning coastal tourism and the escalating numbers of beach visitors. Additionally, the presence of domestic animals like camels and horses used for tourist rides, coupled with an increase in wild animals such as dogs during the lockdown periods, compounded the potential sources of fecal bacteria in the study area. Furthermore, occasional sewage discharge from tourist accommodations and wastewater treatment plants may also contribute to fecal contamination. To effectively mitigate these concerns and bolster public health, a commitment to relentless surveillance efforts, leveraging novel and innovative tools, is essential. These findings underline the crucial interplay between human activities and the health of our coastal ecosystems, emphasizing the need for sustainable practices for a safer and healthier future.

Gastropods as a source for fecal indicator bacteria in drinking water.

Microbial water quality is routinely examined using the fecal indicator bacteria Escherichia coli, coliform bacteria and enterococci. Several practical cases in German drinking water distribution systems indicated invertebrates such as insects or gastropods as a source for the microbiological deterioration. Therefore, we examined three genera of Gastropoda (Arion, Helix and Cepaea) for the presence of fecal indictor bacteria in excreta using standard methods. Enterococci and coliform bacteria were detected in high concentrations (mean values of 1.5 × 106 and 6.3 × 106 per gram feces, respectively). E. coli was also detected, still specification revealed that what was assigned by standard ISO-methods to be E. coli was indeed a novel species of Buttiauxella, exhibiting ß-D-glucuronidase activity, thus, explaining the false-positive results. Microbiome analyses confirmed the cultural results. Enterobacteriaceae were dominant in the samples, yet only very few sequences could be assigned to Escherichia. Our study suggests, that enterococci and coliform bacteria are an integral component in the gastropod microbiome, whereas E. coli might be derived from other sources with gastropods being a vector. The results further indicate, that the current concept of fecal indicator bacteria needs to be extended, as not only humans and homeothermic animals could be a source for fecal indictor bacteria, but also gastropods need to be taken into consideration.

Cross-method assessment of coliphage detection methods in the framework of EPA regulatory standards.

This study aimed to understand the performance and utility of US EPA-approved coliphage methods in comparison to fecal indicator bacteria (FIB) and molecular microbial source tracking (MST) methods in recreational waters. We used US EPA Method 1642 to quantify concentrations of coliphage along with culture- and molecular-based enumeration of E. coli and Enterococcus sp, and human fecal source marker HF183. We also conducted a feasibility assessment to determine the utility of US EPA Method 1642 for application to routine recreational water monitoring. Ten sampling events were conducted at three sampling sites over the duration of a year. Average concentrations of somatic (SC) (log10 1.48 PFU/100 mL) and male-specific (MSC) coliphages (log10 1.00 PFU/100 mL) at all sites were low with SC found across a broader range (0.3-3.1 log10 PFU/100 mL) of concentrations compared to MSC (non-detect-1.7 log10 PFU/100 mL). A feasibility assessment was conducted across US EPA Method 1642 for coliphage enumeration, culture-based FIB, defined substrate technology (DST) approaches Enterolert™ and Colilert®, and quantitative microbial source tracking (qMST) US EPA Method 1696. US EPA Method 1642 had the longest processing times, but also was moderate in cost, compared to the DST and qMST molecular methods. Given the poor correlations between MSC and SC with FIB and qMST markers in this study and the cumbersome nature of US EPA Method 1642, the method may not be the most applicable method for use in systems impacted predominantly by stormwater and other non-point source pollution. Findings from this study, however, provide guidance on the application of fecal indicator virus in ambient coastal surface waters.

Quantitative microbial risk assessment of the gastrointestinal risks to swimmers at Southeast Asian urban beaches using site-specific and combined autochthonous and fecal bacteria exposure data.

Recreational exposure to microbial pollution at urban beaches poses a health risk to beachgoers. The accurate quantification of such risks is crucial in managing beaches effectively and establishing warning guidelines. In this study, we employed a quantitative microbial risk assessment (QMRA) framework to assess marine water quality and estimate the risks associated with Vibrio parahaemolyticus, an autochthonous pathogen that causes gastrointestinal illnesses, and enterococci, a traditional fecal bacteria indicator. The microbial contamination levels of V. parahaemolyticus and enterococci were determined from 48 water samples collected at two beaches in Thailand during dry and wet seasons. The accidentally ingested water volumes were obtained through a survey involving 438 respondents. The probability of illness (Pill) was estimated using dose-response models and Monte Carlo simulation. The results revealed that enterococci posed a higher risk of illness than V. parahaemolyticus at all seven study sites. The median combined gastrointestinal (GI) risk from both bacteria at all sites met the US EPA risk benchmark of 0.036 and the 0.05 benchmark set by the WHO, but the 95th percentile risk data at all sites exceeded the benchmarks. This emphasizes the need for the continuous monitoring and management of microbial pollution at these sites. The site-specific exposure data showed higher estimated risks with increased variations compared to the WHO-referenced values, which highlights the significance of locally measured microbial concentrations and survey exposure data to avoid underestimation. Estimating the risks from recreational exposure to waterborne bacteria can inform beach management policies aimed at reducing public health risks to swimmers. The study findings improve the understanding of the risks associated with water recreation activities at Southeast Asian beaches and offer valuable insights for the development of water quality guidelines, which are crucial for the sustainable development of the blue economy.

Quantification of indicator and pathogenic bacteria in manures and digestates from three agricultural biogas plants over a one-year period.

Interest in the conversion of manure in biogas via anaerobic digestion (AD) is growing, but questions remain about the biosafety of digestates. For a period of one year, we monitored the impact of three mesophilic agricultural biogas plants (BPs) mainly fed with pig manure (BP1, BP3) or bovine manure (BP2) on the physicochemical parameters, the composition of the microbial community and the concentration of bacteria (E. coli, enterococci, Salmonella, Campylobacter, Listeria monocytogenes, Clostridium perfringens, Clostridium botulinum and Clostridioides difficile). The BP2 digestate differed from those of the two other BPs with a higher nitrogen content, more total solids and greater abundance of Clostridia MBA03 and Disgonomonadacea. Persistence during digestion ranked from least to most, was: Campylobacter (1.6 to >2.9 log10 reduction, according to the BP) < E. coli (1.8 to 2.2 log10) < Salmonella (1.1 to 1.4 log10) < enterococci (0.2 to 1.2 log10) and C. perfringens (0.2 to 1 log10) < L. monocytogenes (-1.2 to 1.6 log10) < C. difficile and C. botulinum (≤0.5 log10). No statistical link was found between the reduction in the concentration of the targeted bacteria and the physicochemical and operational parameters likely to have an effect (NH3, volatile fatty acids and total solids contents, hydraulic retention time, presence of co-substrates), underlining the fact that the fate of the bacteria during mesophilic digestion depends on many interacting factors. The reduction in concentrations varied significantly over the sampling period, underlining the need for longitudinal studies to estimate the impact of AD on pathogenic microorganisms.

Environmental risk assessment for fecal contamination sources in urban and peri-urban estuaries, in Escambia and Santa Rosa counties, FL, USA.

Fecal pollution of estuaries and adjacent creeks and streams is of significant concern along the Gulf of Mexico. The prospective threat to human life and water quality impairment via fecal pollution is a substantial danger to the strength and resistance of coastline areas. Pensacola, FL, has a prosperous coastal tourism industry that is utilized for numerous other uses, such as recreational watersports and boating, seafood, and shellfish harvesting. However, the frequency and severity of fecal contamination present possible socio-economic issues, specifically financial hardships. Therefore, understanding the source, abundance, and fate of fecal microbial pollutants in aquatic systems signifies an imperative initial stage for detecting the host sources and techniques to lessen their transport from the landscape. This research aimed to quantify the fecal indicator bacteria (FIB), Escherichia coli, and perform microbiological fecal source tracking to verify if the fecal inputs are of either animal or human host origin. Surface water samples were taken from urban and peri-urban creeks for two sampling periods (February 2021 and January 2022), and IDEXX Colilert-18 (USEPA Standard Method 9223) was used for E. coli enumeration. DNA extractions were obtained from each sample, and quantitative PCR was utilized for fecal microbial source tracking (MST) to detect human, dog, ruminant, and bird host-specific Bacteroides DNA. The result indicates elevated quantities of FIB, E. coli, that surpass the threshold considered safe regarding human health. E. coli at six sites over the two sampling periods exceeded the impairment threshold, reaching as high as 866.4 MPN/100 ml. Fecal source tracking identified human host fecal contamination at four of nine sites, dogs at three of nine, and birds at one site. However, those sites with sources identified via MST all had E. coli levels below impairment thresholds. No sites were determined to be positive for ruminant as a source or for the pathogen Helicobacter pylori. No canine host fecal inputs were found in January 2022, and only one site with human sewage. Our results highlight the utility of MST in assessing bacterial inputs to water bodies and the challenges.

Low antimicrobial resistance in Escherichia coli isolates from two large Austrian alpine karstic spring catchments.

The increasing occurrence of antibiotic resistant bacteria poses a threat to global public health. Clinically relevant resistances also spread through the environment. Aquatic ecosystems in particular represent important dispersal pathways. In the past, pristine water resources have not been a study focus, although ingestion of resistant bacteria through water consumption constitutes a potentially important transmission route. This study assessed antibiotic resistances in Escherichia coli populations in two large well-protected and well-managed Austrian karstic spring catchments representing essential groundwater resources for water supply. E. coli were detected seasonally only during the summer period. By screening a representative number of 551 E. coli isolates from 13 sites in two catchments, it could be shown that the prevalence of antibiotic resistance in this study area is low. 3.4 % of the isolates showed resistances to one or two antibiotic classes, 0.5 % were resistant to three antibiotic classes. No resistances to critical and last-line antibiotics were detected. By integrating fecal pollution assessment and microbial source tracking, we could infer that ruminants were the main hosts for antibiotic resistant bacteria in the studied catchment areas. A comparison with other studies on antibiotic resistances in karstic or mountainous springs highlighted the low contamination status of the model catchments studied here, most likely due to the high protection and careful management while other, less pristine catchments showed much higher antibiotic resistances. We demonstrate that studying easily accessible karstic springs allows a holistic view on large catchments concerning the extent and origin of fecal pollution as well as antibiotic resistance. This representative monitoring approach is also in line with the proposed update of the EU Groundwater Directive (GWD).

Growth and Decay of Fecal Indicator Bacteria and Changes in the Coliform Composition on the Top Surface Sand of Coastal Beaches during the Rainy Season.

High counts of bacteria are present in beach sand, and human health threats attributable to contact with sand have been reported. In this study, we investigated fecal indicator bacteria in the top surface sand of coastal beaches. Monitoring investigations were performed during a monsoon when rainfall occurs randomly, and the composition of the coliforms was analyzed. The coliform count in the top surface sand (depth < 1 cm) increased by approximately 100 fold (26-2.23 × 103 CFU/100 g) with increasing water content because of precipitation. The composition of the coliforms in the top surface sand changed within 24 h of rainfall, with Enterobacter comprising more than 40% of the coliforms. Estimation of factors that changed the bacterial counts and composition revealed that coliform counts tended to increase with increasing water content in the top surface sand. However, the abundance of Enterobacter was independent of the sand surface temperature and water content. Coliform counts in the top surface sand rapidly increased and the composition showed remarkable variations because of the supply of water to the beach following rainfall. Among them, some bacteria with suspected pathogenicity were present. Controlling bacteria in coastal beaches is important for improving public health for beachgoers.

Using watershed characteristics to enhance fecal source identification.

Fecal pollution is one of the most prevalent forms of pollution affecting waterbodies worldwide, threatening public health and negatively impacting aquatic environments. Microbial source tracking (MST) applies polymerase chain reaction (PCR) technology to help identify the source of fecal pollution. In this study, we combine spatial data for two watersheds with general and host-associated MST markers to target human (HF183/BacR287), bovine (CowM2), and general ruminant (Rum2Bac) sources. Concentrations of MST markers in samples were determined with droplet digital PCR (ddPCR). The three MST markers were detected at all sites (n = 25), but bovine and general ruminant markers were significantly associated with watershed characteristics. MST results, combined with watershed characteristics, suggest that streams draining areas with low-infiltration soil groups and high agricultural land use are at an increased risk for fecal contamination. Microbial source tracking has been applied in numerous studies to aid in identifying the sources of fecal contamination, but these studies usually lack information on the involvement of watershed characteristics. Our study combined watershed characteristics with MST results to provide more comprehensive insight into the factors that influence fecal contamination in order to implement the most effective best management practices.