Quantitative polymerase chain reaction (qPCR) offers a rapid, automated, and potentially on-site method for quantifying L. pneumophila in building potable water systems, complementing and potentially replacing traditional culture-based techniques. However, its application in assessing human health risks is complicated by a tendency to overestimate risks due to the detection of genomic copies unassociated with viable, infectious bacteria. This study examines the relationship between L. pneumophila measurements via qPCR and culture-based methods, aiming to establish qPCR-to-culture concentration ratios needed to inform associated health risks. Eligible studies collected quantitative data on L. pneumophila concentrations using molecular and culture-based methods within paired water samples. We developed a Poisson lognormal ratio model and a random-effects meta-analysis model to analyze variations in qPCR-to-culture ratios within and across sites. Of the 17 studies in the systematic review, seven, including 23 site-specific data sets, were used for meta-analysis. Our findings indicate these ratios typically vary from 1:1 to 100:1, with ratios close to 1:1 predicted at all sites. Consequently, adopting a default 1:1 conversion factor appears necessary as a cautious approach to convert qPCR concentrations to culturable concentrations for use in health risk models, such as quantitative microbial risk assessment (QMRA). Where this approach may be too conservative, viability-qPCR could improve the accuracy of qPCR-based QMRA. Standardizing qPCR and culture-based methods and reporting site-specific environmental factors affecting L. pneumophila culturability would improve understanding of the relationship between the two methods. The ratio model introduced here advances beyond simple correlation analyses, facilitating investigations of temporal and spatial heterogeneities in the relationship. This analysis is a step forward in the integration of QMRA and molecular biology, and the framework demonstrated for L. pneumophila is applicable to other pathogens monitored in the environment.

Inhalation of aerosols produced during showering exposes people to chemical and microbial contaminants present in the water. To improve quantitative estimates of exposure and to inform the efficacy of potential interventions to reduce exposures, the number and size distributions of aerosols generated during showering events were monitored and a mass balance model of the generated aerosols was developed. The aerosol generation rates were calculated through calibrating the model with the measured aerosol data. Specifically, aerosol count concentrations and size distributions were measured with an aerodynamic particle sizer over the duration of mock showering events under various conditions, including different water temperatures and different showerhead types (conventional and rain showers). The empirical data were then used to fit a mass balance model to obtain aerosol generation rates and decay rates for each aerosol size class through least square fitting. An initial high peak concentration of aerosols was observed under hot water conditions relative to cold water conditions which resulted in a rapid increase in aerosol exposure during the first 1–2 min of showering. This suggests that people showering in hot water conditions will have a potentially increased exposure during the first 1–2 min. The model-fitted values suggest large inter-experiment variation in estimated aerosol generation and decay rates, even among triplicates of the same showering conditions. Current exposure assessment approaches assume constant aerosol concentrations during showers which might lead to miscalculated cumulative risk for microbial hazards because of their uneven distribution in building plumbing systems and biofilm detachment process during flushing. Thus, considering aerosol dynamics is beneficial during shower exposure assessments to inform risk management interventions. The data set and associated modeling results provided can support this, as they can be readily integrated into microbial risk assessments for waterborne pathogens such as Legionella spp., nontuberculous Mycobacteria (NTM) and Pseudomonas aeruginosa.

Antimicrobial resistance (AMR) poses a global health threat, causing millions of deaths annually, with expectations of increased impact in the future. Wastewater surveillance offers a cost-effective, non-invasive tool to understand AMR carriage trends within a population. We monitored extended-spectrum β-lactamase producing Escherichia coli (ESBL-E. coli) weekly in influent wastewater from six wastewater treatment plants (WWTPs) in Switzerland (November 2021 to November 2022) to investigate spatio-temporal variations, explore correlations with environmental variables, develop a predictive model for ESBL-E. coli carriage in the community, and detect the most prevalent ESBL-genes. We cultured total and ESBL-E. coli in 300 wastewater samples to quantify daily loads and percentage of ESBL-E. coli. Additionally, we screened 234 ESBL-E. coli isolates using molecular methods for the presence of 18 ESBL-gene families. We found a population-weighted mean percentage of ESBL-E. coli of 1.9% (95% confidence interval: 1.8–2%) across all sites and weeks, which can inform ESBL-E. coli carriage. Concentrations of ESBL-E. coli varied across WWTPs and time, with higher values observed in WWTPs serving larger populations. Recent precipitations (previous 24/96 h) showed no significant association with ESBL-E. coli, while temperature occasionally had a moderate impact (P < 0.05, correlation coefficients approximately 0.40) in some locations. We identified blaCTX-M-1, blaCTX-M-9, and blaTEM as the predominant ESBL-gene families. Our study demonstrates that wastewater-based surveillance of culturable ESBL-E. coli provides insights into AMR trends in Switzerland and may also inform resistance. These findings establish a foundation for long term, nationally established monitoring protocols and provide information that may help inform targeted public health interventions.

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.

Greywater reuse is a strategy to address water scarcity, necessitating the selection of treatment processes that balance cost-efficiency and human health risks. A key aspect in evaluating these risks is understanding pathogen contamination levels in greywater, a complex task due to intermittent pathogen occurrences. To address this, faecal indicator organisms like E. coli are often monitored as proxies to evaluate faecal contamination levels and infer pathogen concentrations. However, the wide variability in faecal indicator concentrations poses challenges in their modelling for quantitative microbial risk assessment (QMRA). Our study critically assesses the adequacy of parametric models in predicting the variability in E. coli concentrations in greywater. We found that models that build on summary statistics, like medians and standard deviations, can substantially underestimate the variability in E. coli concentrations. More appropriate models may provide more accurate estimations of, and uncertainty around, peak E. coli concentrations. To demonstrate this, a Poisson lognormal distribution model is fit to a data set of E. coli concentrations measured in shower and laundry greywater sources. This model estimated arithmetic mean E. coli concentrations in laundry waters at approximately 1.0E + 06 MPN 100 mL−1. These results are around 2.0 log10 units higher than estimations from a previously used hierarchical lognormal model based on aggregated summary data from multiple studies. Such differences are considerable when assessing human health risks and setting pathogen reduction targets for greywater reuse. This research highlights the importance of making raw monitoring data available for more accurate statistical evaluations than those based on summary statistics. It also emphasizes the crucial role of model comparison, selection, and validation to inform policy-relevant outcomes.