Large Wastewater Treatment Plant Research Articles

Chemical-Saving Potential for Membrane Bioreactor (MBR) Processes Based on Long-Term Pilot Trials.

Membrane bioreactors (MBRs) have gained attraction in municipal wastewater treatment because of their capacity to meet strict water quality standards and support water reuse. Despite this, their operational sustainability is often compromised by high resource consumption, especially regarding the use of chemicals for membrane cleaning. This study explores innovative membrane-cleaning strategies to enhance the sustainability of MBR processes. Through long-term pilot trials at Stockholm's largest wastewater treatment plant, this study showed that alternative cleaning strategies can reduce chemical use by up to 75% without sacrificing treatment performance. The results further suggest that these alternative strategies could result in cost reductions of up to 70% and a reduction in environmental impacts by as much as 95% for certain indicators. Given that MBRs play a crucial role in addressing increasing treatment demands and advancing circular water management, the outcomes of this study are beneficial for the broader adoption of MBR processes. These results also have implications for existing installations, offering a pathway to more sustainable wastewater treatment. Moreover, the presented cleaning strategies provide significant opportunities for lowering operational costs and reducing the environmental footprint of new and existing MBR installations.

Leveraging wastewater: Validating the national-scale SARS-CoV-2 surveillance system in Cyprus for elevated public health surveillance and enhanced epidemiological insight

The validation of a cross-country, national-scale SARS-CoV-2 wastewater surveillance system for the monitoring of public health in the Republic of Cyprus, took place during October 2021-September 2022. The chronicle of SARS-CoV-2 genetic marker concentrations and prevailing Variants of Concern (VoCs) in all large Wastewater Treatment Plants (WWTPs) of Cyprus for a continuous period of 1 year, was recorded through an integrated, multi-disciplinary sewage-based surveillance system set up to track, visualize and make the collected information and statistical analysis outcomes, publicly available. Composite sampling and physicochemical parameter normalization of Population Equivalents (PE) and SARS-CoV-2 genetic marker concentration loads were utilized.Chemical Oxygen Demand (COD) was selected for normalization of genetic fragment loads. The selection of a normalization method should be tailored according to the characteristics of each individual city/WWTP population structure, flows and wastewater origin. Metagenomic sequencing showed the transition of the SARS-CoV-2 genome from the Delta to the Omicron variant prevalence, in agreement with changes in wastewater gene concentrations. Differences in the early detection of SARS-CoV-2 genetic markers were found among cities. The established system was able to track and monitor the trend of SARS-CoV-2, even in times of absence of other public health measures such as tracing of confirmed COVID-19 cases, showing the capacity of the wastewater system to quickly capture shifts in infections.The work done herein, confirmed the ability of the national-scale system to timely observe pandemic trends, evaluate effects of control measures implemented and monitor the burden of infected patients that shed SARS-CoV-2 fragments.

Treating wastewater for microplastics to a level on par with nearby marine waters

Retention of microplastics (MPs) at the third largest wastewater treatment plant (WWTP) in Sweden was investigated. The plant is one of the most modern and advanced of its kind, with rapid sand filter for tertiary treatment in combination with mechanical, biological, and chemical treatment. It achieved a significantly high treatment efficiency, which brought the MP concentration in its discharge on par with concentrations measured in marine waters of the same region. This novel data shows that properly designed modern WWTPs can reduce the MP content of sewage down to background levels measured in the receiving aquatic environment. Opposite to current understanding of the retention of MP by WWTPs, a modern and well-designed WWTP does not have to be a significant point source for MP. MPs were quantified at all major treatment steps, including digester inlet and outlet sludge. MPs sized 10–500 µm were analyzed by a focal plane array based micro-Fourier transform infrared (FPA-µFTIR) microscopy, a hyperspectral imaging technique, while MPs above 500 µm were analyzed by Attenuated Total Reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Mass was estimated from the hyperspectral images for MPs <500 µm and from microscope images >500 µm. The overall treatment efficiency was in terms of MP counts 99.98 %, with a daily input of 6.42 × 1010 and output of 1.04 × 107 particles. The mass removal efficiency was 99.99 %. The mechanical part of the treatment, the pre-treatment, and primary stages, reduced both the MP counts and mass by approximately 71 %. The combined biological treatment, secondary settling, and final polishing with rapid sand filtration removed nearly all the remaining 29 %. MPs became successively smaller as they passed the different treatment steps. The digester inlet received 1.04 × 1011 MPs daily, while it discharged 9.96 × 1010 MPs, causing a small but not significant decrease in MP counts, with a corresponding MP mass reduction of 9.56 %.

Biochar from recovered cellulose as new admixture in constructed wetlands for micropollutant removal: A circular approach

This study aimed to evaluate the suitability of biochar produced by pyrolysis from recovered wastewater cellulose and activated biologically as an admixture in Constructed Wetlands (CWs) when applied as a post-treatment step to remove micropollutants (MPs) from municipal wastewater effluent. Two planted vertical flow mesocosm CWs with cellulose-based admixtures of different origins (plant residue and recovered toilet paper) were fed with a municipal wastewater effluent representative for rural catchments. The results showed an average MPs elimination of 89.1 % for the activated biochar produced from recovered cellulose when 15 relevant compounds are considered and a reduction of the risk from compounds cocktail below the maximum acceptable level having diclofenac, carbamazepine, PFOS, ciprofloxacin and clarithromycin as main risk drivers (Risk Quotient > 1). The implementation of a circular approach to reduce MPs was finally conducted for the Blies catchment (Saarland region in Germany) characterized by low population density and small, sensitive water bodies. This approach demonstrates the feasibility of combining cellulose recovery with a fine sieve in large wastewater treatment plants (WWTPs) and providing biochar produced from recovered cellulose as an admixture to small WWTP where CW is an affordable solution for MP mitigation.

Using the fish plasma model to evaluate potential effects of pharmaceuticals in effluent from a large urban wastewater treatment plant

Several pharmaceuticals and personal care products (PPCPs) were evaluated using the fish plasma model (FPM) for juvenile Chinook salmon exposed to effluent from a large urban wastewater treatment plant. The FPM compares fish plasma concentrations to therapeutic values determined in human plasma as an indication of potential adverse effects. We used human Cmax values, which are the maximum plasma concentration for a minimum therapeutic dose. Observed and predicted plasma concentrations from juvenile Chinook salmon exposed to a dilution series of whole wastewater effluent were compared to 1%Cmax values to determine Response Ratios (RR) ([plasma]/1%Cmax) for assessment of possible adverse effects. Several PPCPs were found to approach or exceed an RR of 1, indicating potential effects in fish. We also predicted plasma concentrations from measured water concentrations and determined that several of the values were close to or below the analytical reporting limit (RL) indicating potential plasma concentrations for a large number of PPCPs that were below detection. Additionally, the 1%Cmax was less than the RL for several analytes, which could impede predictions of possible effect concentrations. A comparison of observed and predicted plasma concentrations found that observed values were frequently much higher than values predicted with water concentrations, especially for low log10Dow compounds. The observed versus predicted values using the human volume of distribution (Vd), were generally much closer in agreement. These data appear to support the selection of whole-body concentrations to predict plasma values, which relies more on estimating simple partitioning within the fish instead of uptake via water. Overall, these observations highlight the frequently underestimated predicted plasma concentrations and potential to cause adverse effects in fish. Using measured plasma concentrations or predicted values from whole-body concentrations along with improved prediction models and reductions in analytical detection limits will foster more accurate risk assessments of pharmaceutical exposure for fish.

A holistic approach for performance evaluation of wastewater treatment plants: integrating grey water footprint and life cycle impact assessment.

Wastewater treatment plants (WWTPs) have positive and negative impacts on the environment. Therefore, life cycle impact assessment (LCIA) can provide a more holistic framework for performance evaluation than the conventional approach. This study added water footprint (WF) to LCIA and defined ϕ index for accounting for the damage ratio of carbon footprint (CF) to WF. The application of these innovations was verified by comparing the performance of 26 WWTPs. These facilities are located in four different climates in Iran, serve between 1,900 and 980,000 people, and have treatment units like activated sludge, aerated lagoon, and stabilization pond. Here, grey water footprint (GWF) calculated the ecological impacts through typical pollutants. Blue water footprint (BWF) included the productive impacts of wastewater reuse, and CF estimated CO2 emissions from WWTPs. Results showed that GWF was the leading factor. ϕ was 4-7.5% and the average WF of WWTPs was 0.6 m3/ca, which reduced 84%, to 0.1 m³/ca, through wastewater reuse. Here, wastewater treatment and reuse in larger WWTPs, particularly with activated sludge had lower cumulative impacts. Since this method takes more items than the conventional approach, it is recommended for integrated evaluation of WWTPs, mainly in areas where the water-energy nexus is a paradigm for sustainable development.

Metagenomic insights into the wastewater resistome before and after purification at large‑scale wastewater treatment plants in the Moscow city

Wastewater treatment plants (WWTPs) are considered to be hotspots for the spread of antibiotic resistance genes (ARGs). We performed a metagenomic analysis of the raw wastewater, activated sludge and treated wastewater from two large WWTPs responsible for the treatment of urban wastewater in Moscow, Russia. In untreated wastewater, several hundred ARGs that could confer resistance to most commonly used classes of antibiotics were found. WWTPs employed a nitrification/denitrification or an anaerobic/anoxic/oxic process and enabled efficient removal of organic matter, nitrogen and phosphorus, as well as fecal microbiota. The resistome constituted about 0.05% of the whole metagenome, and after water treatment its share decreased by 3–4 times. The resistomes were dominated by ARGs encoding resistance to beta-lactams, macrolides, aminoglycosides, tetracyclines, quaternary ammonium compounds, and sulfonamides. ARGs for macrolides and tetracyclines were removed more efficiently than beta-lactamases, especially ampC, the most abundant ARG in the treated effluent. The removal efficiency of particular ARGs was impacted by the treatment technology. Metagenome-assembled genomes of multidrug-resistant strains were assembled both for the influent and the treated effluent. Ccomparison of resistomes from WWTPs in Moscow and around the world suggested that the abundance and content of ARGs depend on social, economic, medical, and environmental factors.

Enhancing compound confidence in suspect and non-target screening through machine learning-based retention time prediction

The retention time (RT) of contaminants of emerging concern (CECs) in liquid chromatography-high-resolution mass spectrometry (LC-HRMS) is crucial for database matching in non-targeted screening (NTS) analysis. In this study, we developed a machine learning (ML) model to predict RTs of CECs in NTS analysis. Using 1051 CEC standards, we evaluated Random Forest (RF), XGBoost, Support Vector Regression (SVR), and Artificial Neural Network (ANN) with molecular fingerprints and chemical descriptors to establish an optimal model. The SVR model utilizing chemical descriptors resulted in good predictive capacity with R2ext = 0.850 and r2 = 0.925. The model was further validated through laboratory NTS compound characterization. When applied to examine CEC occurrence in a large wastewater treatment plant, we identified 40 level S1 CECs (confirmed structure by reference standard) and 234 level S2 compounds (probable structure by library spectrum match). The model predicted RTs for level S2 compounds, leading to the classification of 153 level S2 compounds with high confidence (ΔRT <2 min). The model served as a robust filtering mechanism within the analytical framework. This study emphasizes the importance of predicted RTs in NTS analysis and highlights the potential of prediction models. Our research introduces a workflow that enhances NTS analysis by utilizing RT prediction models to determine compound confidence levels.

Using Surrogate Parameters to Enhance Monitoring of Community Wastewater Management System Performance for Sustainable Operations.

Community wastewater management systems (CWMS) are small-scale wastewater treatment systems typically in regional and rural areas with less sophisticated treatment processes and often managed by local governments or communities. Research and industrial applications have demonstrated that online UV-Vis sensors have great potential for improving wastewater monitoring and treatment processes. Existing studies on the development of surrogate parameters with models from spectral data for wastewater were largely limited to lab-based. In contrast, industrial applications of these sensors have primarily targeted large wastewater treatment plants (WWTPs), leaving a gap in research for small-scale WWTPs. This paper demonstrates the suitability of using a field-based online UV-Vis sensor combined with advanced data analytics for CWMSs as an early warning for process upset to support sustainable operations. An industry case study is provided to demonstrate the development of surrogate monitoring parameters for total suspended solids (TSSs) and chemical oxygen demand (COD) using the UV-Vis spectral data from an online UV-Vis sensor. Absorbances at a wavelength of 625 nm (UV625) and absorbances at a wavelength of 265 nm (UV265) were identified as surrogate parameters to measure TSSs and COD, respectively. This study contributes to the improvement of WWTP performance with a continuous monitoring system by developing a process monitoring framework and optimization strategy.

Change characteristics, bacteria host, and spread risks of bioaerosol ARGs/MGEs from different stages in sewage and sludge treatment process

The spread of antibiotic resistance genes (ARGs) in the atmospheric environment has seriously threatened human health. Wastewater treatment plants (WWTPs) are an important source of aerosol ARGs. A large WWTP, including sewage treatment process (SWP) and sludge treatment process (SDP), was selected in North China for sampling in this study. The content of ARGs, mobile genetic elements (MGEs), and bacterial genera in sewage/sludge and aerosols from different process stages was detected. The possible correlation between ARGs/ MGEs and bacteria was analyzed. The risk of antibiotic-resistant bacteria was evaluated and the diffusion of ARGs/MGEs was simulated. The results showed that the concentration of ARGs/MGEs varied as the process progressed, and which in the aeration tank was relatively high. The ARGs/MGEs content in SWP aerosol (8.35–163.27 copies/m3) was higher than that in SDP (5.52–16.36 copies/m3). The main ARGs/MGEs detected in SWP aerosol were tnpA-05, tnpA-04, and ermF, while the main ARGs/MGEs detected in SDP aerosol were sul1, ermF, and blaPAO. ARGs were positively correlated with most bacteria and Escherichia coli with ARGs carries higher cytotoxicity. ARGs/MGEs mainly diffused towards the southeast, which may cause harm to urban residents with the diffusion of aerosols. This study provides clues and theoretical basis for preventing the hazards of ARGs from WWTP sources.

Microparticles in Wild and Caged Biota, Sediments, and Water Relative to Large Municipal Wastewater Treatment Plant Discharges.

Anthropogenically modified microparticles including microplastics are present in municipal wastewater treatment plant (WWTP) effluents; however, it is unclear whether biotic exposures are elevated downstream of these outfalls. In the fall of 2019, the present study examined whether microparticle levels in resident fish, environmental samples, and caged organisms were elevated near the Waterloo and Kitchener WWTP outfalls along the Grand River, Ontario, Canada. Wild rainbow darters (Etheostoma caeruleum) were collected from a total of 10 sites upstream and downstream of both WWTPs, along with surface water and sediment samples to assess spatial patterns over an approximately 70-km river stretch. Amphipods (Hyalella azteca), fluted-shell mussels (Lasmigona costata), and rainbow trout (Oncorhynchus mykiss) were also caged upstream and downstream of one WWTP for 14 or 28 days. Whole amphipods, fish digestive tracts, and mussel tissues (hemolymph, digestive glands, gills) were digested with potassium hydroxide, whereas environmental samples were processed using filtration and density separation. Visual identification, measurement, and chemical confirmation (subset only) of microparticles were completed. Elevated abiotic microparticles were found at several upstream reference sites as well as at one or both wastewater-impacted sites. Microparticles in amphipods, all mussel tissues, and wild fish did not show patterns indicative of increased exposures downstream of effluent discharges. In contrast, elevated microparticle counts were found in trout caged directly downstream of the outfall. Across all samples, cellulose fibers (mainly blue and clear colors) were the most common. Overall, results suggest little influence of WWTP effluents on microparticles in biota but rather a ubiquitous presence across most sites that indicates the importance of other point and nonpoint sources to this system. Environ Toxicol Chem 2024;43:1047-1061. © 2024 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Direct application of chemically enhanced primary treatment in a municipal wastewater treatment plant: A case study

A feasible and efficient implementation of an Fe-based chemically enhanced primary treatment (CEPT) in a large conventional wastewater treatment plant (WWTP) was thoroughly evaluated. The influence of the velocity gradient and hydraulic retention time in the coagulation and flocculation stages, as well as the coagulant (FeCl3) dosage, were evaluated at lab-scale. Standard Jar Tests were performed to simulate CEPT, and the results were compared with those obtained by primary settling without coagulant. The increase in FeCl3 dosage and flocculation hydraulic retention time (HRTF) increased the primary settling COD removal capacity between 6.6% and 18.1%, highlighting its effect on the easily biodegradable COD fraction. The COD removal capacity increased up to 16.8% when working with an HRTF of less than 4 min, facilitating CEPT direct application in conventional WWTPs, avoiding the need to incorporate flocculation chamber, with a corresponding decrease in the associated implementation cost. The presence of residual Fe in the primary settling effluent and an increase in the primary sludge volume were the main drawbacks of the process. The coagulation and flocculation velocity gradients or the coagulation hydraulic retention time did not significantly influence the process.

Wastewater-based epidemiology applied at the building-level reveals distinct virome profiles based on the age of the contributing individuals

BackgroundHuman viruses released into the environment can be detected and characterized in wastewater. The study of wastewater virome offers a consolidated perspective on the circulation of viruses within a population. Because the occurrence and severity of viral infections can vary across a person’s lifetime, studying the virome in wastewater samples contributed by various demographic segments can provide valuable insights into the prevalence of viral infections within these segments. In our study, targeted enrichment sequencing was employed to characterize the human virome in wastewater at a building-level scale. This was accomplished through passive sampling of wastewater in schools, university settings, and nursing homes in two cities in Catalonia. Additionally, sewage from a large urban wastewater treatment plant was analysed to serve as a reference for examining the collective excreted human virome.ResultsThe virome obtained from influent wastewater treatment plant samples showcased the combined viral presence from individuals of varying ages, with astroviruses and human bocaviruses being the most prevalent, followed by human adenoviruses, polyomaviruses, and papillomaviruses. Significant variations in the viral profiles were observed among the different types of buildings studied. Mamastrovirus 1 was predominant in school samples, salivirus and human polyomaviruses JC and BK in the university settings while nursing homes showed a more balanced distribution of viral families presenting papillomavirus and picornaviruses and, interestingly, some viruses linked to immunosuppression.ConclusionsThis study shows the utility of building-level wastewater-based epidemiology as an effective tool for monitoring the presence of viruses circulating within specific age groups. It provides valuable insights for public health monitoring and epidemiological studies.

Assessing the determinants of scale effects on carbon efficiency in China's wastewater treatment plants using causal machine learning

The debate over the merits of centralized versus decentralized wastewater treatment plants (WWTPs) has gained prominence considering pressing sustainable development objectives and the need to reduce greenhouse gas (GHG) emissions. This highlights the importance of innovative analytical tools to shape forthcoming policies. Using causal machine learning, we evaluate the impact of WWTP scale on GHG emission intensities and investigate contributing factors. Results show GHG intensity typically decreases as WWTPs scale up. However, this trend varies based on regional environmental, economic, and infrastructure elements. Specifically, regions with fewer industrial wastewater contributions, increased rainwater composition, and elevated temperatures show smaller scale effects. This suggests limited GHG reductions from merely expanding WWTPs in such areas, as the benefits of handling fluctuating inflow volumes, tackling heavy pollution, and operating in cooler conditions offered by larger WWTPs are compromised. This research lays the foundation for comprehensive models promoting sustainable wastewater treatment strategies.

A thorough analysis of the occurrence, removal and environmental risks of organic micropollutants in a full-scale hybrid membrane bioreactor fed by hospital wastewater

The Urban Wastewater Treatment Directive recent draft issued last October 2022 pays attention to contaminants of emerging concern including organic micropollutants (OMPs) and requires the removal of some of them at large urban wastewater treatment plants (WWTPs) calling for their upgrading. Many investigations to date have reported the occurrence of a vast group of OMPs in the influent and many technologies have been tested for their removal at a lab- or pilot-scale. Moreover, it is well-known that hospital wastewater (HWW) contains specific OMPs at high concentration and therefore its management and treatment deserves attention. In this study, a 1-year investigation was carried out at a full-scale membrane bioreactor (MBR) treating mainly HWW. To promote the removal of OMPs, powdered activated carbon (PAC) was added to the bioreactor at 0.1 g/L and 0.2 g/L which resulted in the MBR operating as a hybrid MBR. Its performance was tested for 232 target and 90 non-target OMPs, analyzed by UHPLC-QTOF-MS using a direct injection method. A new methodology was defined to select the key compounds in order to evaluate the performance of the treatments. It was based on their frequency, occurrence, persistence to removal, bioaccumulation and toxicity. Finally, an environmental risk assessment of the OMP residues was conducted by means of the risk quotient approach. The results indicate that PAC addition increased the removal of most of the key OMPs (e.g., sulfamethoxazole, diclofenac, lidocaine) and OMP classes (e.g., antibiotics, psychiatric drugs and stimulants) with the highest loads in the WWTP influent. The hybrid MBR also reduced the risk in the receiving water as the PAC dosage increased mainly for spiramycin, lorazepam, oleandomycin. Finally, uncertainties and issues related to the investigation being carried out at full-scale under real conditions are discussed.

Influenza transmission dynamics quantified from RNA in wastewater in Switzerland.

Influenza infections are challenging to monitor at the population level due to many mild and asymptomatic cases and similar symptoms to other common circulating respiratory diseases, including COVID-19. Methods for tracking cases outside of typical reporting infrastructure could improve monitoring of influenza transmission dynamics. Influenza shedding into wastewater represents a promising source of information where quantification is unbiased by testing or treatment-seeking behaviours. We quantified influenza A and B virus loads from influent at Switzerland's three largest wastewater treatment plants, serving about 14% of the Swiss population (1.2 million individuals). We estimated trends in infection incidence and the effective reproductive number (Re) in these catchments during a 2021/22 epidemic and compared our estimates to typical influenza surveillance data. Wastewater data captured the same overall trends in infection incidence as laboratory-confirmed case data at the catchment level. However, the wastewater data were more sensitive in capturing a transient peak in incidence in December 2021 than the case data. The Reestimated from the wastewater data was roughly at or below the epidemic threshold of 1 during work-from-home measures in December 2021 but increased to at or above the epidemic threshold in two of the three catchments after the relaxation of these measures. The third catchment yielded qualitatively the same results but with wider confidence intervals. The confirmed case data at the catchment level yielded comparatively less precise R_e estimates before and during the work-from-home period, with confidence intervals that included one before and during the work-from-home period. Overall, we show that influenza RNA in wastewater can help monitor nationwide influenza transmission dynamics. Based on this research, we developed an online dashboard for ongoing wastewater-based influenza surveillance in Switzerland.

Removal of microplastic particles during municipal wastewater treatment: A current review

Wastewater treatment plants (WWTPs) are considered the main point source of microplastics (MPs) discharged into water bodies. The article discusses the types and number of MPs present in wastewater and the effectiveness of their removal at the municipal WWTP. The use of different methodologies for determining MPs makes it difficult to compare the results obtained in the studies of different authors. Limitations of various sampling, preparation and final identification techniques of MPs, that may lead to over or underestimating their number in wastewater, were indicated. Although the efficiency of MPs removal on WWPP is reaching 98% or more with advanced treatment, large WWTP could discharged as much as 1010 MPs/day. These values often do not take into account particles smaller than 1 µm (nanoplastics), of which the number in the effluent can be significant. MPs and their degradation products may have a harmful effect on the environment, therefore increasing their removal from wastewater is very important. This can be achieved by introducing additional purification processes, as well as improving the operation of existing ones.

Nitrous Oxide Generation, Influencing Factors and Emission Reduction in Anaerobic-Anoxic-Oxic Process Wastewater Treatment Plant

To achieve the reduction of emissions and control of non-CO2 greenhouse gases in municipal wastewater treatment plants, this study conducted a one-year monitoring of N2O in the A2O process of a large municipal wastewater treatment plant in Beijing. N2O released from the water surface of the A2O process was collected by using a gas flux chamber, and the key influencing factors affecting N2O were analysed using statistics. The results showed that N2O was not easy to be released under non-aerated conditions, and the aerobic zone was its main emission area. Temperature leads to high N2O emissions in winter and low in summer. Correlation analyses showed that NO2-N is one of the most critical factors affecting N2O production, and its accumulation level directly affects the amount of N2O produced in the system. DO also has a great influence on the production of N2O, and the amount of N2O produced decreases with the increase of the DO concentration, and the effect on the enzyme activity also makes the pH have a significant effect on the production of N2O.

Treated municipal wastewater as a source of high-risk and emerging multidrug-resistant clones of E. coli and other Enterobacterales producing extended-spectrum β-lactamases

Extended-spectrum β-lactamase (ESBL)-producing Enterobacterales are a major public health problem, and wastewater from municipal wastewater treatment plants (WWTPs) is a potential means of spreading them into the environment and community. Our objective was to isolate ESBL-producing E. coli and other Enterobacterales from wastewater after treatment at Croatia's largest WWTP and to characterize these isolates by phenotypic and genotypic testing. Of the 200 bacterial isolates, 140 were confirmed as Enterobacterales by MALDI-TOF MS, with Escherichia coli and Klebsiella spp. predominating (69% and 7%, respectively). All 140 enterobacterial isolates were multidrug-resistant (MDR) and produced ESBLs. The most prevalent ESBL genes among the isolates tested were blaCTX-M-15 (60%), blaTEM-116 (44%), and blaCTX-M-3 (13%). Most isolates (94%) carried more than one ESBL gene in addition to blaCTX-M. Genes encoding plasmid-mediated AmpC, most notably blaEBC, were detected in 22% of isolates, whereas genes encoding carbapenemases (blaOXA-48, blaNDM-1, blaVIM-1) were less represented (10%). In E. coli, 9 different sequence types (ST) were found, with the emerging high-risk clones ST361 (serotype A-O9:H30) and pandemic ST131 (serotype B2–O25:H4) predominating (32% and 15%, respectively). Other high-risk E. coli clones included ST405 (3%), ST410 (3%), CC10 (3%), ST10 (3%), and ST38 (2%), and emerging clones included ST1193 (2%) and ST635 (2%). Whole-genome sequencing of three representative E. coli from two dominant clone groups (ST361 and ST131) and one extensively drug-resistant K. oxytoca revealed the presence of multiple plasmids and resistance genes to several other antibiotic classes, as well as association of the blaCTX-M-15 gene with transposons and insertion sequences. Our findings indicate that treated municipal wastewater contributes to the spread of emerging and pandemic MDR E. coli clones and other enterobacterial strains of clinical importance into the aquatic environment, with the risk of reintroduction into humans.

Fate of nitrogen in French human excreta: Current waste and agronomic opportunities for the future

Nitrogen (N) is essential for plant growth and protein synthesis but global reactive N losses, mainly from food systems, induce strong environmental impacts. N losses after human excretion are often overlooked because, in Western societies, they partly occur as inert N2, following denitrification in wastewater treatment plants (WWTP), and losses in waters are often small compared to diffuse agricultural emissions. Yet N from human excretions could be used for crop fertilization, potentially with very high recycling rates via source separation. In this study we use unique operational data from the ∼20,000 French WWTPs to produce a N mass-balance of excretions in the French sanitation system. Even though 75 % of WWTPs' sludge is spread on crops, only 10 % of the excreted N is recycled and 50 % of N is lost to the atmosphere, mainly through WWTP nitrification-denitrification. The remaining 40 % ends up in water or in diffuse losses in the ground, of which about half is lost outside of the WWTPs' discharge system, through sewers storm water and individual autonomous systems. While WWTPs removal efficiency increased in the 2000s, it has been followed by a decade of stagnation, reaching 70 % at the national level. This national average hides regional discrepancies, from 60 to 85 % in the 6 French water agencies basins. These differences closely correlate with the classification as “N sensitive areas” and is mainly due to large WWTPs which handle most of the N load. Recycling all N in excretions could supply 10 % of domestic protein consumption in the current French food system, and up to 30 % if it is prioritized towards crop production for human consumption. Redesigning the food system (decrease of nutrient losses, more plant-based diets) could further increase this contribution.