The shedding of pathogens by infected humans enables the use of sewage monitoring to conduct wastewater-based epidemiology (WBE). Although most WBE studies use data from large sewage treatment plants, timely data from smaller catchments are needed for targeted public health action. Traditional sampling methods, like autosamplers or grab sampling, are not conducive to quick ad hoc deployments and high-resolution monitoring at these smaller scales. This study develops and validates a cheap and easily deployable passive sampler unit, made from readily available consumables, with relevance to the COVID-19 pandemic but with broader use for WBE. We provide the first evidence that passive samplers can be used to detect SARS-CoV-2 in wastewater from populations with low prevalence of active COVID-19 infections (0.034 to 0.34 per 10,000), demonstrating their ability for early detection of infections at three different scales (lot, suburb, and city). A side by side evaluation of passive samplers ( n = 245) and traditionally collected wastewater samples ( n = 183) verified that the passive samplers were sensitive at detecting SARS-CoV-2 in wastewater. On all 33 days where we directly compared traditional and passive sampling techniques, at least one passive sampler was positive when the average SARS-CoV-2 concentration in the wastewater equaled or exceeded the quantification limit of 1.8 gene copies per mL ( n = 7). Moreover, on 13 occasions where wastewater SARS-CoV-2 concentrations were less than 1.8 gene copies per mL, one or more passive samplers were positive. Finally, there was a statistically significant ( p < 0.001) positive relationship between the concentrations of SARS-CoV-2 in wastewater and the levels found on the passive samplers, indicating that with further evaluation, these devices could yield semi-quantitative results in the future. Passive samplers have the potential for wide use in WBE with attractive feasibility attributes of cost, ease of deployment at small-scale locations, and continuous sampling of the wastewater. Further research will focus on the optimization of laboratory methods including elution and extraction and continued parallel deployment and evaluations in a variety of settings to inform optimal use in wastewater surveillance.

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Campylobacter is the leading agent of diarrheal disease worldwide. This study evaluates a novel culture-PCR hybrid (MPN-PCR) assay for the rapid enumeration of Campylobacter spp. from estuarine and wastewater systems. To first evaluate the current, culture-based, Australian standard, an inter-laboratory study was conducted on 69 subsampled water samples. The proposed Most-Probable Number (MPN)-PCR method was then evaluated, by analysing 147 estuarine samples collected over a 2 year period. Data for 14 different biological, hydrological and climatic parameters were also collated to identify pathogen-environment relationships and assess the potential for method specific bias. The results demonstrated that the intra-laboratory performance of the MPN-PCR was superior to that of AS/NZS (σ = 0.7912, P < 0.001; κ = 0.701, P < 0.001) with an overall diagnostic accuracy of ~94%. Furthermore, the analysis of both MPN-PCR and AS/NZS identified the potential for the introduction of method specific bias during assessment of the effects of environmental parameters on Campylobacter spp. numbers.

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At South East Water wastewater treatment plants (WwTPs) in Victoria, Australia, biosolids are stockpiled for three years in compliance with the State guidelines to achieve the highest pathogen reduction grade (T1), suitable for unrestricted use in agriculture and landscaping. However, extended stockpiling is costly, may increase odour nuisance and greenhouse gas emissions, and reduces the fertiliser value of the biosolids. A verification programme of sampling and analysis for enteric pathogens was conducted at two WwTPs where sludge is treated by aerobic and anaerobic digestion, air drying (in drying pans or solar drying sheds) and stockpiling, to enumerate and, if present, monitor the decay of a range of enteric pathogens and parasites. The sludge treatment processes at both WwTPs achieved T1 grade biosolids with respect to prescribed pathogenic bacterial numbers (&lt;1 Salmonella spp. 50 g−1 dry solids (DS) and &lt;100 Escherichia coli g−1 DS) and &gt;3 log10 enteric virus reduction after a storage period of one year. No Ascaris eggs were detected in the influent to the WwTPs, confirming previous studies that the presence of helminth infections in Victoria is extremely low and that Ascaris is not applicable as a control criterion for the microbiological quality of biosolids in the region.