Risk-based critical concentrations of enteric pathogens for recreational water criteria and recommended minimum sample volumes for routine water monitoring.

Concerns are rising about the contamination of recreational waters from human and animal waste, along with associated risks to public health. However, existing guidelines for managing pathogens in these environments have not yet fully integrated risk-based pathogen-specific criteria, which, along with recent advancements in indicators and markers, are essential to improve the protection of public health. This study aimed to establish risk-based critical concentration benchmarks for significant enteric pathogens, i.e., norovirus, rotavirus, adenovirus, Cryptosporidium spp., Giardia lamblia, Campylobacter jejuni, Salmonella spp., and Escherichia coli O157:H7. Applying a 0.036 risk benchmark to both marine and freshwater environments, the study identified the lowest critical concentrations for children, who are the most susceptible group. Norovirus, C. jejuni, and Cryptosporidium presented lowest median critical concentrations for virus, bacteria, and protozoa, respectively: 0.74 GC, 1.73 CFU, and 0.39 viable oocysts per 100 mL in freshwater for children. These values were then used to determine minimum sample volumes corresponding to different recovery rates for culture method, digital polymerase chain reaction and quantitative PCR methods. The results indicate that for children, norovirus required the largest sample volumes of freshwater and marine water (52.08 to 178.57 L, based on the 5th percentile with a 10 % recovery rate), reflecting its low critical concentration and high potential for causing illness. In contrast, adenovirus and rotavirus required significantly smaller volumes (approximately 0.24 to 1.33 L). C. jejuni and Cryptosporidium, which required the highest sampling volumes for bacteria and protozoa, needed 1.72 to 11.09 L and 4.17 to 25.51 L, respectively. Additionally, the presented risk-based framework could provide a model for establishing pathogen thresholds, potentially guiding the creation of extensive risk-based criteria for various pathogens in recreational waters, thus aiding public health authorities in decision-making, strengthening pathogen monitoring, and improving water quality testing accuracy for enhanced health protection.

Greywater recycling for diverse collection scales and appliances: Enteric pathogen log-removal targets and treatment trains.

In light of increasingly diverse greywater reuse applications, this study proposes risk-based log-removal targets (LRTs) to aid the selection of treatment trains for greywater recycling at different collection scales, including appliance-scale reuse of individual greywater streams. An epidemiology-based model was used to simulate the concentrations of prevalent and treatment-resistant reference pathogens (protozoa: Giardia and Cryptosporidium spp., bacteria: Salmonella and Campylobacter spp., viruses: rotavirus, norovirus, adenovirus, and Coxsackievirus B5) in the greywater streams for collection scales of 5-, 100-, and a 1000-people. Using quantitative microbial risk assessment (QMRA), we calculated LRTs to meet a health benchmark of 10-4 infections per person per year over 10'000 Monte Carlo iterations. LRTs were highest for norovirus at the 5-people scale and for adenovirus at the 100- and 1000-people scales. Example treatment trains were designed to meet the 95 % quantiles of LRTs. Treatment trains consisted of an aerated membrane bioreactor, chlorination, and, if required, UV disinfection. In most cases, rotavirus, norovirus, adenovirus and Cryptosporidium spp. determined the overall treatment train requirements. Norovirus was most often critical to dimension the chlorination (concentration × time values) and adenovirus determined the required UV dose. Smaller collection scales did not generally allow for simpler treatment trains due to the high LRTs associated with viruses, with the exception of recirculating washing machines and handwashing stations. Similarly, treating greywater sources individually resulted in lower LRTs, but the lower required LRTs nevertheless did not generally allow for simpler treatment trains. For instance, LRTs for a recirculating washing machine were around 3-log units lower compared to LRTs for indoor reuse of combined greywater (1000-people scale), but both scenarios necessitated treatment with a membrane bioreactor, chlorination and UV disinfection. However, simpler treatment trains may be feasible for small-scale and application-scale reuse if: (i) less conservative health benchmarks are used for household-based systems, considering the reduced relative importance of treated greywater in pathogen transmission in households, and (ii) higher log-removal values (LRVs) can be validated for unit processes, enabling simpler treatment trains for a larger number of appliance-scale reuse systems.

Quantitative Risk Assessment of Wind-Supported Transmission of Highly Pathogenic Avian Influenza Virus to Dutch Poultry Farms via Fecal Particles from Infected Wild Birds in the Environment.

A quantitative microbial risk assessment model was developed to estimate the probability that the aerosolization of fecal droppings from wild birds in the vicinity of poultry farms would result in the infection of indoor-housed poultry with highly pathogenic avian influenza virus (HPAIv) in the Netherlands. Model input parameters were sourced from the scientific literature and experimental data. The availability of data was diverse across input parameters, and especially parameters on the aerosolization of fecal droppings, survival of HPAIv and dispersal of aerosols were uncertain. Model results indicated that the daily probability of infection of a single poultry farm is very low, with a median value of 7.5 × 10-9. Accounting for the total number of poultry farms and the length of the bird-flu season, the median overall probability of at least one HPAIv-infected poultry farm during the bird-flu season is 2.2 × 10-3 (approximately once every 455 years). This is an overall estimate, averaged over different farm types, virus strains and wild bird species, and results indicate that uncertainty is relatively high. Based on these model results, we conclude that it is unlikely that this introduction route plays an important role in the occurrence of HPAIv outbreaks in indoor-housed poultry.

Risk of Illness with Salmonella Spp. and Antibiotic-Resistant Salmonella sp. due to Consumption of Lettuce Irrigated with Water from La Ramada Irrigation District.

Despite heavy contamination of the Bogotá River with domestic and industrial waste, it remains vital for various purposes, including agricultural use at La Ramada Irrigation District. There are important concerns regarding pathogen concentrations in irrigation water at La Ramada, including the presence of antibiotic-resistant Salmonella spp. This study aimed to estimate the risk of Salmonella-related illness from consuming lettuce irrigated with Bogotá River water at La Ramada. We collected lettuce samples from 4 different sites, all irrigated with water from La Ramada. The methodology involved a process to detach Salmonella spp. from lettuce leaves, quantification through plate counts on SS agar, and establishment of antibiotic-resistant bacteria concentrations through growth on media supplemented with ampicillin or ciprofloxacin. The results showed concentrations of Salmonella spp. of 103.59,102.66, and 104.56 CFU/g lettuce at sites 1, 2, and 3, respectively, and ampicillin-resistant Salmonella spp. of 101.93, 101.31, and 102.07 CFU/g lettuce at sites 1, 2, and 3, respectively. No colonies were obtained from lettuce samples collected from site 4. Notably, we detected no isolates resistant to ciprofloxacin at any of the sites. Salmonella spp. concentrations varied greatly among sampling sites. Salmonella spp. concentrations were used to predict the daily probability of illness, with a probability of 0.59 (0.33 to 0.78, CI 95%) for Salmonella spp. and 0.3 (0.03 to 0.53, CI 95%) for ampicillin-resistant Salmonella spp.

Evolution and health risk of indicator microorganisms in landscape water replenished by reclaimed water.

As an important means to solve water shortage, reclaimed water has been widely used for landscape water supply. However, with the emergence of large-scale epidemic diseases such as SARS, avian influenza and COVID-19 in recent years, people are increasingly concerned about the public health safety of reclaimed water discharged into landscape water, especially the pathogenic microorganisms in it. In this study, the water quality and microorganisms of the Old Summer Palace, a landscape water body with reclaimed water as the only replenishment water source, were tracked through long-term dynamic monitoring. And the health risks of indicator microorganisms were analyzed using Quantitative Microbial Risk Assessment (QMRA). It was found that the concentration of indicator microorganisms Enterococcus (ENT), Escherichia coli (EC) and Fecal coliform (FC) generally showed an upward trend along the direction of water flow and increased by more than 0.6 log at the end of the flow. The concentrations of indicator microorganisms were higher in summer and autumn than those in spring. And there was a positive correlation between the concentration of indicator microorganisms and COD. Further research suggested that increased concentration of indicator microorganisms also led to increased health risks, which were more than 30% higher in other areas of the park than the water inlet area and required special attention. In addition, (water) surface operation exposure pathway had much higher health risks than other pathways and people in related occupations were advised to take precautions to reduce the risks.

Bioaerosols emission from source facilities in a wastewater treatment plant: Critical exposure time and sensitivity analysis.

Overexposure of sewage workers to bioaerosol released from wastewater treatment plants (WWTPs) can cause serious infections, but practical method for controlling their health risk is lacking. In this study, reverse quantitative microbial risk assessment was used to estimate the daily critical exposure time (CET) of sewage workers exposing to Staphylococcus aureus bioaerosol emitted by three emission sources facilities in a WWTP based on either U.S. EPA or WHO benchmark, and sensitivity analysis was conducted to analyze the influence of various parameters on the outcomes of CET. The results showed that the CET of females was always 1.12-1.29 times that of males. In addition, the CET after wearing face masks was 28.28-52.37 times as long as before. The working time can be determined based on the CET results of male workers wearing face masks exposed to the inverted-umbrella aeration tank (14.73-550.98 min for U.S. EPA benchmark and 55.07-1972.24 min for WHO benchmark). In each scenario, the variable parameter exposure concentration (ec) always showed the most influence on the CET results. After wearing the face masks, the removal fraction by employing face masks also had a significant effect on the results, only second to ec. Therefore, the wearing of face mask is the most convenient and effective measure to prolong the CET. Furthermore, practical methods to reducing bioaerosol concentration in WWTPs exposure are also necessary to extend CET and safeguard worker health. This study enriches the application range of reverse quantitative microbial risk assessment framework and provides theoretical support for stakeholders to establish reasonable working time threshold guidelines, and practical method and novel perspective to protect the on-site health risks of sewage workers exposing to various facilities.

Quantitative microbial risk assessment (QMRA) of the work of manual pit emptiers, commonly known as bayakous.

In Haiti, manual pit emptiers, known as bayakous, face significant health risks. They work by descending naked into latrine pits, exposing themselves to pathogens and contributing to environmental contamination. This study employs the quantitative microbial risk assessment (QMRA) method to evaluate the microbial risks associated with this practice, considering nine prevalent pathogens in Haiti. Three ingestion scenarios were developed: hand-to-mouth contact, ingestion while immersed in excreta, and a combination of both. A sensitivity analysis assessed the impact of input data on study outcomes. The results indicate a high probability of infection and illness during pit emptying operations annually for all scenarios and pathogens. Recommendations include adopting personal protective equipment (PPE) and using a manual Gulper waste pump to eliminate the need to descend directly into the pits, thereby reducing the risk of injury from sharp objects. The study proposes the establishment of intermediate disposal points approximately 5 km from collection sites to deter illegal dumping. National regulations and professionalization of the bayakou profession are suggested, along with awareness campaigns to promote PPE and Gulper pump usage. Addressing these issues is crucial for safeguarding the health of bayakou and public health in Haiti.

Quantitative microbial risk assessment for Escherichia Coli O157: H7 via drinking water in the Gaza Strip, Palestine.

Introduction: Microbial contamination of drinking water, particularly by pathogens such as Escherichia coli O157: H7, is a significant public health concern worldwide, especially in regions with limited access to clean water like the Gaza Strip. However, few studies have quantified the disease burden associated with E. coli O157: H7 contamination in such challenging water management contexts. Objective: This study aimed to conduct a comprehensive Quantitative Microbial Risk Assessment to estimate the annual infection risk and disease burden attributed to E. coli O157: H7 in Gaza's drinking water. Methods: Applying the typical four steps of the Quantitative Microbial Risk Assessment technique-hazard identification, exposure assessment, dose-response analysis, and risk characterization-the study assessed the microbial risk associated with E. coli O157: H7 contamination in Gaza's drinking water supply. A total of 1317 water samples from various sources across Gaza were collected and analyzed for the presence of E. coli O157: H7. Using Microsoft ExcelTM and @RISKTM software, a Quantitative Microbial Risk Assessment model was constructed to quantify the risk of infection associated with E. coli O157: H7 contamination. Monte Carlo simulation techniques were employed to assess uncertainty surrounding input variables and generate probabilistic estimates of infection risk and disease burden. Results: Analysis of the water samples revealed the presence of E. coli O157: H7 in 6.9% of samples, with mean, standard deviation, and maximum values of 1.97, 9.74, and 112 MPN/100 ml, respectively. The risk model estimated a median infection risk of 3.21 × 10-01 per person per year and a median disease burden of 3.21 × 10-01 Disability-Adjusted Life Years per person per year, significantly exceeding acceptable thresholds set by the WHO. Conclusion: These findings emphasize the urgent need for proactive strategies to mitigate public health risks associated with waterborne pathogens in Gaza.

Two risk assessments: Evaluating the use of indicator HF183 Bacteroides versus pathogen measurements for modelling recreational illness risks in an urban watershed.

The purpose of this study was to evaluate the performance of HF183 Bacteroides for estimating pathogen exposures during recreational water activities. We compared the use of Bacteroides-based exposure assessment to exposure assessment that relied on pathogen measurements. We considered two types of recreational water sites: those impacted by combined sewer overflows (CSOs) and those not impacted by CSOs. Samples from CSO-impacted and non-CSO-impacted urban creeks were analysed by quantitative polymerase chain reaction (qPCR) for HF183 Bacteroides and eight human gastrointestinal pathogens. Exposure assessment was conducted two ways for each type of site (CSO-impacted vs. non-CSO impacted): 1) by estimating pathogen concentrations from HF183 Bacteroides concentrations using published ratios of HF183 to pathogens in sewage and 2) by estimating pathogen concentrations from qPCR measurements. QMRA (quantitative microbial risk assessment) was then conducted for swimming, wading, and fishing exposures. Overall, mean risk estimates varied from 0.27 to 53 illnesses per 1,000 recreators depending on exposure assessment, site, activity, and norovirus dose-response model. HF183-based exposure assessment identified CSO-impacted sites as higher risk, and the recommended HF183 risk-based threshold of 525 genomic copies per 100 mL was generally protective of public health at the CSO-impacted sites but was not as protective at the non-CSO-impacted sites. In the context of our urban watershed, HF183-based exposure assessment over- and under-estimated risk relative to exposure assessment based on pathogen measurements, and the etiology of predicted pathogen-specific illnesses differed significantly. Across all sites, the HF183 model overestimated risk for norovirus, adenovirus, and Campylobacter jejuni, and it underestimated risk for E. coli and Cryptosporidium. To our knowledge, this study is the first to directly compare health risk estimates using HF183 and empirical pathogen measurements from the same waterways. Our work highlights the importance of site-specific hazard identification and exposure assessment to decide whether HF183 is applicable for monitoring risk.

Risk Assessment Predicts Most of the Salmonellosis Risk in Raw Chicken Parts is Concentrated in Those Few Products with High Levels of High-Virulence Serotypes of Salmonella.

Salmonella prevalence declined in U.S. raw poultry products since adopting prevalence-based Salmonella performance standards, but human illnesses did not reduce proportionally. We used Quantitative Microbial Risk Assessment (QMRA) to evaluate public health risks of raw chicken parts contaminated with different levels of all Salmonella and specific high- and low-virulence serotypes. Lognormal Salmonella level distributions were fitted to 2012 USDA-FSIS Baseline parts survey and 2023 USDA-FSIS HACCP verification sampling data. Three different Dose-Response (DR) approaches included (i) a single DR for all serotypes, (ii) DR that reduces Salmonella Kentucky ST152 virulence, and (iii) multiple serotype-specific DR models. All scenarios found risk concentrated in the few products with high Salmonella levels. Using a single DR model with Baseline data (µ = -3.19, σ = 1.29 Log CFU/g), 68% and 37% of illnesses were attributed to the 0.7% and 0.06% of products with >1 and >10 CFU/g Salmonella, respectively. Using distributions from 2023 HACCP data (µ = -5.53, σ = 2.45), 99.8% and 99.0% of illnesses were attributed to the 1.3% and 0.4% of products with >1 and >10 CFU/g Salmonella, respectively. Scenarios with serotype-specific DR models showed more concentrated risk at higher levels. Baseline data showed 92% and 67% and HACCP data showed >99.99% and 99.96% of illnesses attributed to products with >1 and >10 CFU/g Salmonella, respectively. Regarding serotypes using Baseline or HACCP input data, 0.002% and 0.1% of illnesses were attributed to the 0.2% and 0.4% of products with >1 CFU/g of Kentucky ST152, respectively, while 69% and 83% of illnesses were attributed to the 0.3% and 0.6% of products with >1 CFU/g of Enteritidis, Infantis, or Typhimurium, respectively. Therefore, public health risk in chicken parts is concentrated in finished products with high levels and specifically high levels of high-virulence serotypes. Low-virulence serotypes like Kentucky contribute few human cases.

A secondary model for the effect of pH on the variability in growth fitness of Listeria innocua strains.

Variability in microbial growth is a keystone of modern Quantitative Microbiological Risk Assessment (QMRA). However, there are still significant knowledge gaps on how to model variability, with the most common assumption being that variability is constant. This is implemented by an error term (with constant variance) added on top of the secondary growth model (for the square root of the growth rate). However, this may go against microbial ecology principles, where differences in growth fitness among bacterial strains would be more prominent in the vicinity of the growth limits than at optimal growth conditions. This study coins the term "secondary models for variability", evaluating whether they should be considered in QMRA instead of the constant strain variability hypothesis. For this, 21 strains of Listeria innocua were used as case study, estimating their growth rate by the two-fold dilution method at pH between 5 and 10. Estimates of between-strain variability and experimental uncertainty were obtained for each pH using mixed-effects models, showing the lowest variability at optimal growth conditions, increasing towards the growth limits. Nonetheless, the experimental uncertainty also increased towards the extremes, evidencing the need to analyze both sources of variance independently. A secondary model was thus proposed, relating strain variability and pH conditions. Although the modelling approach certainly has some limitations that would need further experimental validation, it is an important step towards improving the description of variability in QMRA, being the first model of this type in the field.

Site-specific risk-based threshold (RBT) concentrations for sewage-associated markers in estuarine swimming waters.

This study establishes site-specific risk-based threshold (RBT) concentrations for sewage-associated markers, including Bacteroides HF183 (HF183), Lachnospiraceae Lachno3 (Lachno3), cross-assembly phage (CrAssphage), and pepper mild mottle virus (PMMoV), utilizing quantitative microbial risk assessment (QMRA) for recreational estuarine waters (EW). The QMRA model calculates a RBT concentration corresponding to a selected target illness risk for ingestion of EW contaminated with untreated sewage. RBT concentrations were estimated considering site-specific decay rates and concentrations of markers and reference pathogen (human norovirus; HNoV), aiding in the identification of high-risk days during the swimming season. Results indicated varying RBT concentrations for fresh (Day 0) and aged (Days 1 to 10) sewage contamination scenarios over 10 days. HF183 exhibited the highest RBT concentration (26,600 gene copis (GC)/100 mL) initially but decreased rapidly with aging (2570 to 3120 GC/100 mL on Day 10) depending on the decay rates, while Lachno3 and CrAssphage remained relatively stable. PMMoV, despite lower initial RBT (3920 GC/100 mL), exhibited increased RBT (4700 to 6440 GC/100 mL) with aging due to its slower decay rate compared to HNoV. Sensitivity analysis revealed HNoV concentrations as the most influential parameter. Comparison of marker concentrations in estuarine locations with RBT concentrations showed instances of marker exceedance, suggesting days of potential higher risks. The observed discrepancies between bacterial and viral marker concentrations in EW highlight the need for optimized sample concentration method and simultaneous measurement of multiple markers for enhanced risk predictions. Future research will explore the utility of multiple markers in risk management. Overall, this study contributes to better understanding human health risks in recreational waters, aiding regulators, and water quality managers in effective decision-making for risk prioritization and mitigation strategies.

Estimation and evaluation of the risks of protozoa infections associated to the water from a treatment plant in southern Brazil using the Quantitative Microbiological Risk Assessment Methodology (QMRA).

In this study, the Quantitative Microbial Risk Assessment (QMRA) methodology was applied to estimate the annual risk of Giardia and Cryptosporidium infection associated with a water treatment plant in southern Brazil. The efficiency of the treatment plant in removing protozoa and the effectiveness of the Brazilian legislation on microbiological protection were evaluated, emphasizing the relevance of implementing the QMRA in this context. Two distinct approaches were employed to estimate the mechanical removal of protozoa: The definitions provided by the United States Environmental Protection Agency (USEPA), and the model proposed by Neminski and Ongerth. Although the raw water collected had a higher concentration of Giardia cysts than Cryptosporidium oocysts, the estimated values for the annual risk of infection were significantly higher for Cryptosporidium than for Giardia. From a general perspective, the risk values of protozoa infection were either below or very near the limit set by the World Health Organization (WHO). In contrast, all the risk values of Cryptosporidium infection exceeded the threshold established by the USEPA. Ultimately, it was concluded that the implementation of the QMRA methodology should be considered by the Brazilian authorities, as the requirements and guidelines provided by the Brazilian legislation proved to be insufficient to guarantee the microbiological safety of drinking water. In this context, the QMRA application can effectively contribute to the prevention and investigation of outbreaks of waterborne disease.

Integrating quantitative microbiological risk assessment and disability-adjusted life years to evaluate the effects of urbanization on health risks for river recreationists.

Urban rivers provide an excellent opportunity for water recreation. This study probabilistically assessed health risks associated with water recreation in urban rivers in the Bitan Scenic Area, Taiwan, by employing quantitative microbial risk assessment and disability-adjusted life years (DALYs). Moreover, the effects of urbanization on the health risks of river recreation induced by waterborne pathogenic Escherichia coli (E. coli) were investigated. First, data on river E. coli levels were collected in both the Bitan Scenic Area and the upstream river section, and model parameters were obtained through a questionnaire administered to river recreationists. Monte Carlo simulation was then employed to address parameter uncertainty. Finally, DALYs were calculated to quantify the cumulative effects in terms of potential life lost and years lived with disability. The results indicated that the 90 % confidence intervals for the disease burden (DB) were 0.2-74.1 × 10-6, 0.01-94.0 × 10-6, and 0.3-128.9 × 10-6 DALY per person per year (pppy) for canoeing, swimming, and fishing, respectively, in the Bitan Scenic Area. Furthermore, urbanization near the Bitan Scenic Area approximately doubled the DB risks to river recreationists in upstream rural areas. At the 95th percentile, the DB risks exceeded the tolerances recommended by the World Health Organization (1 × 10-6) or U.S. Environmental Protection Agency (1 × 10-4). The findings suggest that the simultaneous implementation of effluent sewer systems and best management practices can reduce health risks to river recreationists by at least half, reducing the DALY levels below 1 × 10-4 or even 1 × 10-5 pppy.

Quantitative Microbial Risk Assessment Framework Incorporating Water Ages with Legionella pneumophila Growth Rates.

Water age in drinking water systems is often used as a proxy for water quality but is rarely used as a direct input in assessing microbial risk. This study directly linked water ages in a premise plumbing system to concentrations of Legionella pneumophila via a growth model. In turn, the L. pneumophila concentrations were used for a quantitative microbial risk assessment to calculate the associated probabilities of infection (Pinf) and clinically severe illness (Pcsi) due to showering. Risk reductions achieved by purging devices, which reduce water age, were also quantified. The median annual Pinf exceeded the commonly used 1 in 10,000 (10-4) risk benchmark in all scenarios, but the median annual Pcsi was always 1-3 orders of magnitude below 10-4. The median annual Pcsi was lower in homes with two occupants (4.7 × 10-7) than with one occupant (7.5 × 10-7) due to more frequent use of water fixtures, which reduced water ages. The median annual Pcsi for homes with one occupant was reduced by 39-43% with scheduled purging 1-2 times per day. Smart purging devices, which purge only after a certain period of nonuse, maintained these lower annual Pcsi values while reducing additional water consumption by 45-62%.

Quantitative microbial risk assessment for ingestion of antibiotic resistance genes from private wells contaminated by human and livestock fecal sources.

We used quantitative microbial risk assessment to estimate ingestion risk for intI1, erm(B), sul1, tet(A), tet(W), and tet(X) in private wells contaminated by human and/or livestock feces. Genes were quantified with five human-specific and six bovine-specific microbial source-tracking (MST) markers in 138 well-water samples from a rural Wisconsin county. Daily ingestion risk (probability of swallowing ≥1 gene) was based on daily water consumption and a Poisson exposure model. Calculations were stratified by MST source and soil depth over the aquifer where wells were drilled. Relative ingestion risk was estimated using wells with no MST detections and >6.1 m soil depth as a referent category. Daily ingestion risk varied from 0 to 8.8 × 10-1 by gene and fecal source (i.e., human or bovine). The estimated number of residents ingesting target genes from private wells varied from 910 (tet(A)) to 1,500 (intI1 and tet(X)) per day out of 12,000 total. Relative risk of tet(A) ingestion was significantly higher in wells with MST markers detected, including wells with ≤6.1 m soil depth contaminated by bovine markers (2.2 [90% CI: 1.1-4.7]), wells with >6.1 m soil depth contaminated by bovine markers (1.8 [1.002-3.9]), and wells with ≤6.1 m soil depth contaminated by bovine and human markers simultaneously (3.1 [1.7-6.5]). Antibiotic resistance genes (ARGs) were not necessarily present in viable microorganisms, and ingestion is not directly associated with infection. However, results illustrate relative contributions of human and livestock fecal sources to ARG exposure and highlight rural groundwater as a significant point of exposure.IMPORTANCEAntibiotic resistance is a global public health challenge with well-known environmental dimensions, but quantitative analyses of the roles played by various natural environments in transmission of antibiotic resistance are lacking, particularly for drinking water. This study assesses risk of ingestion for several antibiotic resistance genes (ARGs) and the class 1 integron gene (intI1) in drinking water from private wells in a rural area of northeast Wisconsin, United States. Results allow comparison of drinking water as an exposure route for antibiotic resistance relative to other routes like food and recreational water. They also enable a comparison of the importance of human versus livestock fecal sources in the study area. Our study demonstrates the previously unrecognized importance of untreated rural drinking water as an exposure route for antibiotic resistance and identifies bovine fecal material as an important exposure factor in the study setting.

Transmission of Viruses from Restroom Use: A Quantitative Microbial Risk Assessment.

Restroom use has been implicated in a number of viral outbreaks. In this study, we apply quantitative microbial risk assessment to quantify the risk of viral transmission by contaminated restroom fomites. We estimate risk from high-touch fomite surfaces (entrance/exit door, toilet seat) for three viruses of interest (SARS-CoV-2, adenovirus, norovirus) through eight exposure scenarios involving differing user behaviors, and the use of hand sanitizer following each scenario. We assessed the impacts of several sequences of fomite contacts in the restroom, reflecting the variability of human behavior, on infection risks for these viruses. Touching of the toilet seat was assumed to model adjustment of the seat (open vs. closed), a common touch point in single-user restrooms (home, small business, hospital). A Monte Carlo simulation was conducted for each exposure scenario (10,000 simulations each). Norovirus resulted in the highest probability of infection for all exposure scenarios with fomite surfaces. Post-restroom automatic-dispensing hand sanitizer use reduced the probability of infection for each virus by up to 99.75%. Handwashing within the restroom, an important risk-reduction intervention, was not found to be as effective as use of a non-touch hand sanitizer dispenser for reducing risk to near or below 1/1,000,000, a commonly used risk threshold for comparison.

Occurrence, transmission and risks assessment of pathogens in aquatic environments accessible to humans.

Pathogens are ubiquitously detected in various natural and engineered water systems, posing potential threats to public health. However, it remains unclear which human-accessible waters are hotspots for pathogens, how pathogens transmit to these waters, and what level of health risk associated with pathogens in these environments. This review collaboratively focuses and summarizes the contamination levels of pathogens on the 5 water systems accessible to humans (natural water, drinking water, recreational water, wastewater, and reclaimed water). Then, we showcase the pathways, influencing factors and simulation models of pathogens transmission and survival. Further, we compare the health risk levels of various pathogens through Quantitative Microbial Risk Assessment (QMRA), and assess the limitations of water-associated QMRA application. Pathogen levels in wastewater are consistently higher than in other water systems, with no significant variation for Cryptosporidium spp. among five water systems. Hydraulic conditions primarily govern the transmission of pathogens into human-accessible waters, while environmental factors such as temperature impact pathogens survival. The median and mean values of computed public health risk levels posed by pathogens consistently surpass safety thresholds, particularly in the context of recreational waters. Despite the highest pathogens levels found in wastewater, the calculated health risk is significantly lower than in other water systems. Except pathogens concentration, variables like the exposure mode, extent, and frequency are also crucial factors influencing the public health risk in water systems. This review shares valuable insights to the more accurate assessment and comprehensive management of public health risk in human-accessible water environments.

Evaluation of virus removal in membrane bioreactor (MBR) and conventional activated sludge (CAS) processes based on long-term monitoring at two wastewater treatment plants.

The membrane bioreactor (MBR) process always offers better wastewater treatment than conventional activated sludge (CAS) treatment. However, the difference in their efficacy of virus reduction remains unknown. To investigate this, we monitored virus concentrations before and after MBR and CAS processes over 2 years. Concentrations of norovirus genotypes I and II (NoV GI and GII), aichivirus (AiV), F-specific RNA phage genotypes I, II, and III (GI-, GII-, and GIII-FRNAPHs), and pepper mild mottle virus (PMMoV) were measured by a quantitative polymerase chain reaction (qPCR) method at two municipal wastewater treatment plants (WWTPs A and B) in Japan. Virus concentration datasets containing left-censored data were estimated by using both maximum likelihood estimation (MLE) and robust regression on order statistics (rROS) approaches. PMMoV was the most prevalent at both WWTPs, with median concentrations of 7.5 to 8.8 log10 copies/L before treatment. Log10 removal values (LRVs) of all viruses based on means and standard deviations of concentrations before and after treatment were consistently higher following MBR than following CAS. We used NoV GII as a model pathogen in a quantitative microbial risk assessment of the treated water, and we estimated the additional reductions required following MBR and CAS processes to meet the guideline of 10-6 DALYs pppy for safe wastewater reuse.

Quantitative assessment of food safety interventions for Campylobacter spp. and Salmonella spp. along the chicken meat supply chain in Burkina Faso and Ethiopia.

Rural and small-scale chicken farming is a major source of income in most African countries, and chicken meat is an important source of nutrients. However, chicken meat can be contaminated with Campylobacter spp. and Salmonella spp., pathogens with a high reported burden of foodborne illnesses. Therefore, it is essential to control these pathogens in chicken meat. Quantitative microbial risk assessments (QMRA) can aid the development of effective food safety control measures and are currently lacking in chicken meat supply chains in the African context. In this study, we developed stochastic QMRA models for Salmonella spp. and Campylobacter spp. in the chicken meat supply chain in Burkina Faso and Ethiopia employing the modular process risk model in @Risk software. The study scope covered chicken farming, transport, slaughtering, consumer handling, and consumption. Effectiveness of candidate interventions was assessed against baseline models' outputs, which showed that the mean annual Campylobacter spp. risk estimates were 6482 cases of illness per 100,000 persons and 164 disability adjusted life years (DALYs) per 100,000 persons in Burkina Faso, and 12,145 cases and 272 DALYs per 100,000 persons in Ethiopia. For Salmonella spp., mean annual estimates were 2713 cases and 1212 DALYs per 100,000 persons in Burkina Faso, and 4745 cases and 432 DALYs per 100,000 persons in Ethiopia. Combining interventions (improved hand washing plus designated kitchen utensils plus improved cooking) resulted in 75 % risk reduction in Burkina Faso at restaurants and 93 to 94 % in Ethiopia at homes for both Salmonella spp. and Campylobacter spp. For Burkina Faso, adding good hygienic slaughter practices at the market to these combined interventions led to over 91 % microbial risk reduction. Interventions that involved multiple food safety actions in a particular step of the supply chain or combining different interventions from different steps of the supply chain resulted in more risk reduction than individual action interventions. Overall, this study demonstrates how diverse and scanty food supply chain information can be applied in QMRA to provide estimates that can be used to stimulate risk-based food safety action in African countries.