Despite high vaccination rates in the Netherlands, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to circulate. Longitudinal sewage surveillance was implemented along with the notification of cases as two parts of the surveillance pyramid to validate the use of sewage for surveillance, as an early warning tool, and to measure the effect of interventions. Sewage samples were collected from nine neighborhoods between September 2020 and November 2021. Comparative analysis and modeling were performed to understand the correlation between wastewater and case trends. Using high resolution sampling, normalization of wastewater SARS-CoV-2 concentrations, and ‘normalization’ of reported positive tests for testing delay and intensity, the incidence of reported positive tests could be modeled based on sewage data, and trends in both surveillance systems coincided. The high collinearity implied that high levels of viral shedding around the onset of disease largely determined SARS-CoV-2 levels in wastewater, and that the observed relationship was independent of variants of concern and vaccination levels. Sewage surveillance alongside a large-scale testing effort where 58 % of a municipality was tested, indicated a five-fold difference in the number of SARS-CoV-2-positive individuals and reported cases through standard testing. Where trends in reported positive cases were biased due to testing delay and testing behavior, wastewater surveillance can objectively display SARS-CoV-2 dynamics for both small and large locations and is sensitive enough to measure small variations in the number of infected individuals within or between neighborhoods. With the transition to a post-acute phase of the pandemic, sewage surveillance can help to keep track of re-emergence, but continued validation studies are needed to assess the predictive value of sewage surveillance with new variants. Our findings and model aid in interpreting SARS-CoV-2 surveillance data for public health decision-making and show its potential as one of the pillars of future surveillance of (re)emerging viruses.

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The ecological state of receiving water bodies can be significantly influenced by organic micropollutants that are emitted via stormwater runoff. Reported efforts to quantify the emission of micropollutants mainly focus on sampling at combined sewer overflows and storm sewer outfalls, which can be challenging. An alternative method, called fingerprinting, was developed and tested in this study. The fingerprinting method utilizes wastewater treatment plant (WWTP) influent samples and derives the proportion of stormwater in a sample. This is achieved by comparing the wet weather vs dry weather concentrations of substances-tracers which are present only in wastewater. It is then possible to estimate the concentration of organic micropollutants in stormwater runoff from measurements in the influent of a WWTP based on a mass balance. In this research, the fingerprinting method was applied in influent samples obtained in five WWTPs in the Netherlands. In total, 28 DWF and 22 WWF samples were used. The chosen tracers were ibuprofen, 2-hydroxyibuprofen, naproxen and diclofenac. Subsequently, the concentration in stormwater runoff of 403 organic micropollutants was estimated via the WWF samples. The substances that were present and analyzed included glyphosate and AMPA, 24 out of 254 pesticides, 6 out of 28 organochlorine pesticides, 45 out of 63 pharmaceuticals, 15 out of 15 PAHs, 2 of the 7 PCBs, and 20 of 33 other substances (e.g. bisphenol-A). A comparison with findings from other studies suggested that the fingerprinting method yields trustworthy results. It was also noted that a representative and stable dry weather flow reference concentration is a strict requirement for the successful application of the proposed method.

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Monitoring of SARS-CoV-2 in wastewater (WW) is a promising tool for epidemiological surveillance, correlating not only viral RNA levels with the infection dynamics within the population, but also to viral diversity. However, the complex mixture of viral lineages in WW samples makes tracking of specific variants or lineages circulating in the population a challenging task. We sequenced sewage samples of 9 WW-catchment areas within the city of Rotterdam, used specific signature mutations from individual SARS-CoV-2 lineages to estimate their relative abundances in WW and compared them against those observed in clinical genomic surveillance of infected individuals between September 2020 and December 2021. We showed that especially for dominant lineages, the median of the frequencies of signature mutations coincides with the occurrence of those lineages in Rotterdam's clinical genomic surveillance. This, along with digital droplet RT-PCR targeting signature mutations of specific variants of concern (VOCs), showed that several VOCs emerged, became dominant and were replaced by the next VOC in Rotterdam at different time points during the study. In addition, single nucleotide variant (SNV) analysis provided evidence that spatio-temporal clusters can also be discerned from WW samples. We were able to detect specific SNVs in sewage, including one resulting in the Q183H amino acid change in the Spike gene, that was not captured by clinical genomic surveillance. Our results highlight the potential use of WW samples for genomic surveillance, increasing the set of epidemiological tools to monitor SARS-CoV-2 diversity.@en

Illicit connections of wastewater to stormwater systems are the main drawback of separate sewer systems, as they lead to a direct discharge of untreated wastewater to the aquatic environment. Consequently, several inspection methods have been developed for detecting illicit connections. This study simultaneously applied several low- and high-tech methods for the detection of illicit connections in the same catchment (De Heuvel, the Netherlands). The methods included mesh wire screens for capturing coarse contamination, measurements of electroconductivity and temperature, sampling and quantification of Escherichia coli and extended-spectrum ß-lactamase-producing E. coli (ESBL-EC), DNA analysis via quantitative polymerase chain reaction for human-, dog-, and bird-specific fecal indicators, and distributed temperature sensing. Significant illicit connections could be identified using all methods. Nonetheless, hydraulic conditions and, predominantly, the sewage volume determine whether a misconnection can be detected by especially the low-tech methods. Using these results, the identified misconnections were repaired and biological and DNA analyses were repeated. Our results demonstrate that there were no changes in E. coli or ESBL-EC before and after mitigation, suggesting that these common markers of fecal contamination are not specific enough to evaluate the performance of mitigation efforts. However, a marked decrease in human wastewater markers (HF183) was observed.

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Drinking water distribution networks (WDNs) are a crucial infrastructure for life in cities. Deterioration of this ageing, and partly hidden from view, infrastructure can result in losses due to leakage and an increased contamination risk. To counteract this, maintenance strategies are required to maintain the service level. Information on the most critical elements of a WDN, with respect to the functioning of the system as a whole, is essential for prioritising maintenance or rehabilitation activities. In this study a Graph theory based method is developed and applied for efficiently identifying the most critical elements. The main advantage of this method is that it avoids the need to perform elaborate hydrodynamic model calculations. Instead, the structure of the network is the main starting point. The results show that the structure of the network is more decisive than the hydraulics with respect to the criticality of the system’s performance as a whole. Results depict that the suggested approach is applicable not only to the main (primary) network, but also to the capillaries which are normally beyond the scope of the traditional methods applied so-far because of the complexity of the networks and the required calculation time.

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