Wastewater-based epidemiology (WBE) is a great approach that enables us to comprehensively monitor the community to determine the scale and dynamics of infections in a city, particularly in metropolitan cities with a high population density. Therefore, we monitored the time course of the SARS-CoV-2 RNA concentration in raw sewage in the Frankfurt metropolitan area, the European financial center. To determine the SARS-CoV-2 RNA concentration in sewage, we continuously collected 24 h composite samples twice a week from two wastewater treatment plant (WWTP) influents (Niederrad and Sindlingen) serving the Frankfurt metropolitan area and performed RT-qPCR analysis targeting three genes (N gene, S gene, and ORF1ab gene). In August, a resurgence in the SARS-CoV-2 RNA load was observed, reaching 3 × 1013 copies/day, which represented similar levels compared to April with approx. 2 × 1014 copies/day. This corresponds to a continuous increase again in COVID-19 cases in Frankfurt since August, with an average of 28.6 incidences, compared to 28.7 incidences in April. Different temporal dynamics were observed between different sampling points, indicating local dynamics in COVID-19 cases within the Frankfurt metropolitan area. The SARS-CoV-2 RNA load to the WWTP Niederrad ranged from approx. 4 × 1011 to 1 × 1015 copies/day, the load to the WWTP Sindlingen from approx. 1 × 1011 to 2 × 1014 copies/day, which resulted in a preceding increase in these loading in July ahead of the weekly averaged incidences. The study shows that WBE has the potential as an early warning system for SARS-CoV-2 infections and a monitoring system to identify global hotspots of COVID-19.

This study investigated the potential of Membrane-Aerated Biofilm Reactors (MABRs) for mainstream nitrogen removal via partial nitration/anaerobic ammonium oxidation (anammox). Four laboratory-scale MABRs were operated with real municipal wastewater characterized by low concentrations of nitrogen (varying between 31 and 120 mg–NH 4 –N L −1 ) and the presence of biodegradable organic carbon (soluble COD (sCOD) between 7 and 230 mg-O 2 L −1 ). Two reactors were operated with different aeration strategies (intermittent vs. continuous), the other two with differences in biomass retention (recirculation or removal of detached biomass). Keeping a constant HRT caused instabilities due to difficulties with setting the optimal oxygen flux for the respective surface loadings (1.6–6 g–NH 4 –N m −2 d −1 ). Operating the MABRs with a constant surface loading (2 g–NH 4 –N m −2 d −1 ) resulted in higher and more stable total nitrogen (TN) removal independent of the aeration strategy. The intermittently aerated MABR improved from an average TN removal of 23%–69%, the continuously aerated MABR from 20% to 50% TN removal. Independent of the feeding strategy, the continuously aerated reactor removed slightly more ammonium (80–95%) compared to the intermittently aerated reactor (74–93%). Limiting the oxygen supply by intermittent aeration proofed successful to favor partial nitritation and anammox. Continuous aeration did not achieve stable suppression of nitrite oxidizing bacteria (NOB). Of the removed ammonium, approx. 26% were left in the effluent as nitrate (only 10% with intermittent aeration). Recirculation of the detached biomass resulted in reattachment onto the biofilm or membrane surface. This recirculation led to significantly higher biomass retention times and thus to better performance. Removing detached biofilm from the reactor caused a slightly lower TN removal of 33% compared to 45% with reattachment, while average ammonium removal was 58% compared to 63%, respectively. Scouring events had a significant impact on the overall operation, resulting in short term losses of TN removal capacities of 50–100%. The microbial community composition was different depending on the aeration strategy and biomass retention. The continuously aerated reactor contained significantly more AOB than the intermittently aerated MABR. The reactor with biomass retention contained less ammonium oxidizing bacteria (AOB), compared to the reactor with low biomass retention. In all MABRs, anammox bacteria established in the biofilm after an initial drop in abundance.

For community-level monitoring, the European Commission under the EU Sewage Sentinel System recommends wastewater-based SARS-CoV-2 surveillance. Tracking SARS-CoV-2 variants in a community is pivotal for appropriate public health response. Genome sequencing of SARS-CoV-2 in wastewater samples for tracking variants is challenging, often resulting in low coverage genome sequences, thereby impeding the detection of the SARS-CoV-2 mutations. Therefore, we aimed at high-coverage SARS-CoV-2 genome sequences from sewage samples which we successfully accomplished. This first pan-European surveillance compared the mutation profiles associated with the variants of concerns: B.1.1.7, P.1, B.1.351 and B.1.617.2 across 20 European countries, including 54 municipalities. The results highlight that SARS-CoV-2 variants detected in the wastewater samples mirror the variants profiles reported in clinical data. This study demonstrated that >98% coverage of SARS-CoV-2 genomic sequences is possible and can be used to track SARS-CoV-2 mutations in wastewater to support identifying variants circulating in a city at the community level.

Wastewater-based severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) surveillance of Frankfurt Airport by genome sequencing was used to detect SARS-CoV-2 variants entering the region. In November 2021, we found all characteristic mutations of Omicron in wastewater originating from Frankfurt Airport before the first confirmed clinical report from an arriving passenger on 26 November 2021.

We report a sequencing analysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in wastewater samples collected in the Frankfurt, Germany, metropolitan area. The majority of the detected mutations have been identified only in clinical genomes outside Frankfurt, indicating that the sequencing of SARS-CoV-2 RNA in wastewater can provide insights into emerging variants in a city.