Assessment of seasonality and normalization techniques for wastewater-based surveillance in Ontario, Canada.

Introduction: Wastewater-based surveillance is at the forefront of monitoring for community prevalence of COVID-19, however, continued uncertainty exists regarding the use of fecal indicators for normalization of the SARS-CoV-2 virus in wastewater. Using three communities in Ontario, sampled from 2021-2023, the seasonality of a viral fecal indicator (pepper mild mottle virus, PMMoV) and the utility of normalization of data to improve correlations with clinical cases was examined. Methods: Wastewater samples from Warden, the Humber Air Management Facility (AMF), and Kitchener were analyzed for SARS-CoV-2, PMMoV, and crAssphage. The seasonality of PMMoV and flow rates were examined and compared by Season-Trend-Loess decomposition analysis. The effects of normalization using PMMoV, crAssphage, and flow rates were analyzed by comparing the correlations to clinical cases by episode date (CBED) during 2021. Results: Seasonal analysis demonstrated that PMMoV had similar trends at Humber AMF and Kitchener with peaks in January and April 2022 and low concentrations (troughs) in the summer months. Warden had similar trends but was more sporadic between the peaks and troughs for PMMoV concentrations. Flow demonstrated similar trends but was not correlated to PMMoV concentrations at Humber AMF and was very weak at Kitchener (r = 0.12). Despite the differences among the sewersheds, unnormalized SARS-CoV-2 (raw N1-N2) concentration in wastewater (n = 99-191) was strongly correlated to the CBED in the communities (r = 0.620-0.854) during 2021. Additionally, normalization with PMMoV did not improve the correlations at Warden and significantly reduced the correlations at Humber AMF and Kitchener. Flow normalization (n = 99-191) at Humber AMF and Kitchener and crAssphage normalization (n = 29-57) correlations at all three sites were not significantly different from raw N1-N2 correlations with CBED. Discussion: Differences in seasonal trends in viral biomarkers caused by differences in sewershed characteristics (flow, input, etc.) may play a role in determining how effective normalization may be for improving correlations (or not). This study highlights the importance of assessing the influence of viral fecal indicators on normalized SARS-CoV-2 or other viruses of concern. Fecal indicators used to normalize the target of interest may help or hinder establishing trends with clinical outcomes of interest in wastewater-based surveillance and needs to be considered carefully across seasons and sites.

Wild fish responses to wastewater treatment plant upgrades in the Grand River, Ontario.

Municipal wastewater treatment plant (WWTP) effluent is one of several point sources of contaminants (nutrients, pharmaceuticals, estrogens, etc.) which can lead to adverse responses in aquatic life. Studies of WWTP effluent impacts on rainbow darter (Etheostoma caeruleum) collected downstream of WWTPs in the Grand River, Ontario have reported disruption at multiple levels of biological organization, including altered vitellogenin gene expression, lower levels of in vitro steroid production, and high frequency of intersex. However, major upgrades have occurred at treatment plants in the central Grand River over the last decade. Treatment upgrades to the Waterloo WWTP were initiated in 2009 but due to construction delays, the upgrades came fully on-line in 2017/2018. Responses in rainbow darter have been followed at sites associated with the outfall consistently over this entire time period. The treatment plant upgrade resulted in nitrification of effluent, and once complete there was a major reduction in effluent ammonia, selected pharmaceuticals, and estrogenicity. This study compared several key responses in rainbow darter associated with the Waterloo WWTP outfall prior to and post upgrades. Stable isotopes signatures in fish were used to track exposure to effluent and changed dramatically over time, corresponding to the effluent quality. Disruptions in in vitro steroid production and intersex in the darters that had been identified prior to the upgrades were no longer statistically different from the upstream reference sites after the upgrades. Although annual variations in water temperature and flow can potentially mask or exacerbate the effects of the WWTP effluent, major capital investments in wastewater treatment targeted at improving effluent quality have corresponded with the reduction of adverse responses in fish in the receiving environment.

River metabolic fingerprints and regimes reveal ecosystem responses to enhanced wastewater treatment.

Although many studies have examined how improvements in wastewater treatment impact river nutrient concentrations and loads, there has been much less focus on measuring river metabolism to evaluate the wider aquatic ecosystem benefits of reducing nutrient inputs to rivers. The objectives of this study were to evaluate the effects of enhanced wastewater treatment (nitrification) on river metabolism in the Grand River, Canada's largest river draining into Lake Erie. Metabolic fingerprints and regimes (calculated from high-frequency dissolved oxygen [DO] measurements) were used to visualize whole-river ecosystem functional responses to these wastewater treatment upgrades. There was a 60% reduction in ecosystem respiration during summer, in response to reductions in effluent total ammonia inputs, causing a shift from net heterotrophy to net autotrophy, and contraction of river metabolic fingerprints. This resulted in major improvements in summer DO concentrations, with reductions in the percentage of days during summer that DO minima fell below water-quality guidelines for protection of aquatic early life stages, from 88% to ≤16%. The results also point to potential cascading impacts on coupled phosphorus and nitrogen cycles, which may generate further improvements in river water quality. During the summer, high rates of river metabolism and nutrient retention may result in measured water-column nutrient concentrations potentially underestimating nutrient pressures. This study also demonstrates the value of combining river metabolism with nutrient monitoring for a more holistic understanding of the role of nutrients in river ecosystem health and function.

Assessment of the use of mainstream iron addition for phosphorous control on H2 S content of biogas from anaerobic digestion of sludges.

A material flux analysis on sulfur (S), phosphorus (P), aluminum (Al), and iron (Fe) was conducted for two WWTPs (Galt and Kitchener) to evaluate the potential of coagulants that are employed for phosphorus control to reduce hydrogen sulfide (H2 S) emissions in the biogas from anaerobic digestion. It was found that while the Galt WWTP receives higher concentrations of S in the raw wastewater than the Kitchener WWTP, this had only a modest impact on the speciation of S entering anaerobic digestion. At both plants, only 2%-4% of influent S entered the digesters. The presence of Fe in the sludge stream was found to cause S, that is released by volatile solid destruction and sulfate ( SO 4 2 - SO 4 2 - ) reduction, to become particulate-bound. A dosage of 1.1 mg/L of Fe into the raw wastewater (11% of the Fe dosed for P control) was sufficient for sulfide (S2- ) control. Transitioning the Galt WWTP from Al to Fe dosing for P control had no significant impact on effluent P concentrations and resulted in a substantial reduction in the biogas H2 S concentration. An additional secondary benefit was an increase in the solid content of the dewatered cake. PRACTITIONER POINTS: Material flux analyses can be employed to gain insight into the fate of key elements contributing to biogas quality. The use of iron for phosphorous control can effectively control H2 S in anaerobic biogas. Conversion from Al2 (SO4 )3 to FeSO4 dosing for P control resulted in increased solid content of centrifuged biosolids.

Impact of anaerobically digested biosolids characteristics and handling conditions on dewatering performance at multiple facilities.

Anaerobically digested biosolids (ABD) characteristics that affect dewatering were assessed at three water resource recovery facilities (WRRF) with different handling practices. Dewatering performance at the three sites corresponded to different levels of soluble chemical oxygen demand (COD), ammonia (NH4 -N), and mono- and divalent cation concentrations in ADB. Capillary suction time (CST) and a modified centrifugal technique were used to determine optimum polymer doses and to assess the impact of handling conditions on dewatering performance. Both techniques indicated that polymer dosing between 15 and 20 kg/dry tonne was optimal for all facilities and that biosolids mixing and pumping did not significantly impact dewaterability. The CST values of anaerobically digested biosolids decreased as temperature increased, but no significant difference was found for either temperature or location of dewatering facilities. Sludge viscosity and rheological properties that vary with temperature appeared to have influenced CST values. Modified centrifugal technique results indicated cake solids were not affected by polymer make-up water or ADB temperature when emulsion polymer was used. This study shows the value of laboratory testing of biosolids under controlled conditions to identify and correct potential problems in full-scale operations. PRACTITIONER POINTS: Capillary suction time and a modified centrifugal technique were used to assess the impact of different process-related and environmental factors on dewatering. Higher concentrations of soluble COD (potentially extracellular polymeric substances - EPS) and low calcium (Ca) in anaerobically digested biosolids align with reduced dewaterability. Cell disruption and break down of floc structures due to storage/mixing and pumping of biosolids did not appear to negatively impact dewatering. Modified centrifugal test results did not provide conclusive evidence of whether dewatering of anaerobically digested biosolids could be significantly impacted by temperature over the range 15-30°C, especially when emulsion polymer is used. This study shows the value of laboratory testing of biosolids under controlled conditions to identify potential problems in the full-scale operations.

Autotrophic nitrogen-removing biofilms on porous and non-porous membranes.

The effective removal of nitrogen compounds from wastewater has become a critical issue for treatment plants as the awareness of their negative impact on the environment increased. Autotrophic nitrogen removal has become an interesting alternative to the more conventional heterotrophic processes, as it eliminates the need for an organic carbon addition to the source water and reduces biomass yields. Gas transfer membrane biofilm reactors (MBfR) for nitrification and hydrogen driven denitrification are of special interest as they combine membrane diffusers and biofilms, provide an efficient supply of necessary electron donor for autotrophic removal of ammonia and nitrate, extend solids retention times and retain biomass within the reactor. Subsequently, a wide range of MBfR, which vary based on the type of membrane material and membrane module configuration, are being tested for this purpose. This paper reviews the research to date and also discusses the challenges that still lay ahead before MBfR can be used at treatment plants.