This paper presents risk-based enteric pathogen log reduction targets for non-potable and potable uses of a variety of alternative source waters (i.e., locally-collected greywater, roof runoff, and stormwater). A probabilistic Quantitative Microbial Risk Assessment (QMRA) was used to derive the pathogen log 10 reduction targets (LRTs) that corresponded with an infection risk of either 10 −4 per person per year (ppy) or 10 −2 ppy. The QMRA accounted for variation in pathogen concentration and sporadic pathogen occurrence (when data were available) in source waters for reference pathogens in the genera Rotavirus, Mastadenovirus(human adenoviruses), Norovirus, Campylobacter, Salmonella, Giardia and Cryptosporidium. Non-potable uses included indoor use (for toilet flushing and clothes washing) with occasional accidental ingestion of treated nonpotable water (or cross-connection with potable water), and unrestricted irrigation for outdoor use. Various exposure scenarios captured the uncertainty from key inputs, i.e., the pathogen concentration in source water; the volume of water ingested; and for the indoor use, the frequency of and the fraction of the population exposed to accidental ingestion. Both potable and non-potable uses required pathogen treatment for the selected waters and the LRT was generally greater for potable use than non-potable indoor use and unrestricted irrigation. The difference in treatment requirements among source waters was driven by the microbial quality of the water-both the density and occurrence of reference pathogens. Greywater from collection systems with 1000 people had the highest LRTs; however, those for greywater collected from a smaller population (~ 5 people), which have less frequent pathogen occurrences, were lower. Stormwater had highly variable microbial quality, which resulted in a range of possible treatment requirements. The microbial quality of roof runoff, and thus the resulting LRTs, remains uncertain due to lack of relevant pathogen data.

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We propose a minimum set of meta-information to accompany the reporting of SARS-CoV-2 occurrence in wastewater for improved data interpretation.

Risk-based treatment of onsite wastewaters for decentralized reuse requires information on the occurrence and density of pathogens in source waters, which differ from municipal wastewater due to scaling and dilution effects in addition to variable source contributions. In this first quantitative report of viral enteric pathogens in onsite-collected graywater and wastewater, untreated graywater (n = 50 samples) and combined wastewater (i.e., including blackwater; n = 28) from three decentralized collection systems were analyzed for two norovirus genogroups (GI/GII) and human adenoviruses using droplet digital polymerase chain reaction (ddPCR). Compared to traditional quantitative PCR (qPCR), which had insufficient sensitivity to quantify viruses in graywater, ddPCR allowed quantification of norovirus GII and adenovirus in 4% and 14% of graywater samples, respectively (none quantifiable for norovirus GI). Norovirus GII was routinely quantifiable in combined wastewater by either PCR method (96% of samples), with wellcorrelated results between the analyses (R 2 = 0.96) indicating a density range of 5.2-7.9 log 10 genome copies/L. These concentrations are greater than typically reported in centralized municipal wastewater, yet agree well with an epidemiology-based model previously used to develop pathogen log-reduction targets (LRTs) for decentralized non-potable water systems. Results emphasize the unique quality of onsite wastewaters, supporting the previous LRTs and further quantitative microbial risk assessment (QMRA) of decentralized water reuse.

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As decentralized water reuse continues to gain popularity, risk-based treatment guidance is increasingly sought for the protection of public health. However, effort s to evaluate pathogen risks and log-reduction requirements have been hindered by an incomplete understanding of pathogen occurrence and densities in locally-collected wastewaters (i.e., from decentralized collection systems). Of particular interest is the potentially high enteric pathogen concentration in small systems with an active infected excreter, but generally lower frequency of pathogen occurrences in smaller systems compared to those with several hundred contributors. Such variability, coupled with low concentrations in many source streams (e.g., sink, shower/bath, and laundry waters), has limited direct measurement of pathogens. This study presents an approach to modeling pathogen concentrations in variously sized greywater and combined wastewater collection systems based on epidemiological pathogen incidence rates, user population size, and fecal loadings to various residential wastewater sources. Pathogen infections were modeled within various population sizes (5-, 100-, and 1,000-person) for seven reference pathogens (viruses: adenoviruses, Norovirus, and Rotavirus; bacteria: Campylobacter and Salmonella spp.; and protozoa: Cryptosporidium and Giardia spp.) on each day of 10,000 possible years, accounting for intermittent infection and overlap of infection periods within the population. Fecal contamination of fresh greywaters from bathroom sinks, showers/baths, and laundry, as well as combined greywater and local combined wastewater (i.e., including toilets), was modeled based on reported fecal indicators in the various sources. Simulated daily infections and models of fecal contamination were coupled with pathogen shedding characteristics to generate distributions of pathogen densities in the various waters. The predicted frequency of pathogen occurrences in local wastewaters was generally low due to low infection incidence within small cohort groups, but increased with collection scale (population size) and infection incidence rate (e.g., Norovirus). When pathogens did occur, a decrease in concentrations from 5- to 100- and from 100- to 1,000-person systems was observed; nonetheless, overall mean concentrations (i.e., including non-occurrences) remained the same due to the increased number of occurrences. This highlights value of the model for characterizing scaling effects over averaging methods, which overestimate the frequency of pathogen occurrence in small systems while underestimating concentration peaks that likely drive risk periods. Results of this work will inform development of risk-based pathogen reduction requirements for decentralized water reuse.

Listeria monocytogenes is detected by electrochemical impedance spectroscopy using a mouse monoclonal antibody immobilized onto an Au electrode. This yields sensitivities of 0.825 kΩ cm2/(CFU/mL) and 1.129 kΩ cm2/(CFU/mL) and detection limits of 5 CFU/mL and 4 CFU/mL for ideal solutions and filtered tomato extract, respectively. Control experiments with an Au electrode onto which a mouse monoclonal antibody to GAPDH is immobilized demonstrate that non‐specific adsorption is insignificant for the system and methodology studied here. Control experiments with Salmonella enterica demonstrate no cross‐reactivity to this food pathogen. Potential technological hurdles to development of a multiplexed impedance biosensor for food pathogens are also discussed.

In this study, we report the human health risk of gastrointestinal infection associated with inhalation exposure to airborne zoonotic pathogens emitted following application of dairy cattle manure to land. Inverse dispersion modeling with the USEPA's AERMOD dispersion model was used to determine bioaerosol emission rates based on edge-of-field bioaerosol and source material samples analyzed by real-time quantitative polymerase chain reaction (qPCR). Bioaerosol emissions and transport simulated with AERMOD, previously reported viable manure pathogen contents, relevant exposure pathways, and pathogen-specific dose-response relationships were then used to estimate potential downwind risks with a quantitative microbial risk assessment (QMRA) approach. Median 8-h infection risks decreased exponentially with distance from a median of 1:2700 at edge-of-field to 1:13 000 at 100 m and 1:200 000 at 1000 m; peak risks were considerably greater (1:33, 1:170, and 1:2500, respectively). These results indicate that bioaerosols emitted from manure application sites following manure application may present significant public health risks to downwind receptors. Manure management practices should consider improved controls for bioaerosols in order to reduce the risk of disease transmission.