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- COST Action ES1403: New and Emerging challenges and opportunities in wastewater REUSe (NEREUS)Publication . Fatta-Kassinos, Despo; Manaia, C.; Berendonk, T. U.; Cytryn, E.; Bayona, J.; Chefetz, B.; Slobodnik, J.; Kreuzinger, N.; Rizzo, L.; Malato, S.; Lundy, L.; Ledin, A.
- Urban wastewater treatment plants as hotspots for the release of antibiotics in the environment: a reviewPublication . Michael, I.; Rizzo, L.; McArdell, C. S.; Manaia, C. M.; Merlin, C.; Schwartz, T.; Dagot, C.; Fatta-Kassinos, D.Urban wastewater treatment plants (UWTPs) are among the main sources of antibiotics' release into various compartments of the environment worldwide. The aim of the present paper is to critically review the fate and removal of various antibiotics in wastewater treatment, focusing on different processes (i.e. biological processes, advanced treatment technologies and disinfection) in view of the current concerns related to the induction of toxic effects in aquatic and terrestrial organisms, and the occurrence of antibiotics that may promote the selection of antibiotic resistance genes and bacteria, as reported in the literature. Where available, estimations of the removal of antibiotics are provided along with the main treatment steps. The removal efficiency during wastewater treatment processes varies and is mainly dependent on a combination of antibiotics' physicochemical properties and the operating conditions of the treatment systems. As a result, the application of alternative techniques including membrane processes, activated carbon adsorption, advanced oxidation processes (AOPs), and combinations of them, which may lead to higher removals, may be necessary before the final disposal of the effluents or their reuse for irrigation or groundwater recharge.
- Performance of secondary wastewater treatment methods for the removal of contaminants of emerging concern implicated in crop uptake and antibiotic resistance spread: a reviewPublication . Krzeminski, Pawel; Tomei, Maria Concetta; Karaolia, Popi; Langenhoff, Alette; Almeida, C. Marisa R.; Felis, Ewa; Gritten, Fanny; Andersen, Henrik Rasmus; Fernandes, Telma; Manaia, Célia M.; Rizzo, Luigi; Fatta-Kassinos, DespoContaminants of emerging concern (CEC) discharged in effluents of wastewater treatment plants (WWTPs), not specifically designed for their removal, pose serious hazards to human health and ecosystems. Their impact is of particular relevance to wastewater disposal and re-use in agricultural settings due to CEC uptake and accumulation in food crops and consequent diffusion into the food-chain. This is the reason why the chemical CEC discussed in this review have been selected considering, besides recalcitrance, frequency of detection and entity of potential hazards, their relevance for crop uptake. Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have been included as microbial CEC because of the potential of secondary wastewater treatment to offer conditions favourable to the survival and proliferation of ARB, and dissemination of ARGs. Given the adverse effects of chemical and microbial CEC, their removal is being considered as an additional design criterion, which highlights the necessity of upgrading conventional WWTPs with more effective technologies. In this review, the performance of currently applied biological treatment methods for secondary treatment is analysed. To this end, technological solutions including conventional activated sludge (CAS), membrane bioreactors (MBRs), moving bed biofilm reactors (MBBRs), and nature-based solutions such as constructed wetlands (CWs) are compared for the achievable removal efficiencies of the selected CEC and their potential of acting as reservoirs of ARB&ARGs. With the aim of giving a picture of real systems, this review focuses on data from full-scale and pilot-scale plants treating real urban wastewater. To achieve an integrated assessment, technologies are compared considering also other relevant evaluation parameters such as investment and management costs, complexity of layout and management, present scale of application and need of a post-treatment. Comparison results allow the definition of design and operation strategies for the implementation of CEC removal in WWTPs, when agricultural reuse of effluents is planned.
- High throughput analysis of integron gene cassettes in wastewater environmentsPublication . Gatica, Joao; Tripathi, Vijay; Green, Stefan; Manaia, Célia M.; Berendonk, Thomas; Cacace, Damiano; Merlin, Christophe; Kreuzinger, Norbert; Schwartz, Thomas; Fatta-Kassinos, Despo; Rizzo, Luigi; Schwermer, Carsten U.; Garelick, Hemda; Jurkevitch, Edouard; Cytryn, EddieIntegrons are extensively targeted as a proxy for anthropogenic impact in the environment. We developed a novel high-throughput amplicon sequencing pipeline that enables characterization of thousands of integron gene cassette-associated reads, and applied it to acquire a comprehensive overview of gene cassette composition in effluents from wastewater treatment facilities across Europe. Between 38 100 and 172 995 reads per-sample were generated and functionally characterized by screening against nr, SEED, ARDB and β-lactamase databases. Over 75% of the reads were characterized as hypothetical, but thousands were associated with toxin-antitoxin systems, DNA repair, cell membrane function, detoxification and aminoglycoside and β-lactam resistance. Among the reads characterized as β-lactamases, the carbapenemase blaOXA was dominant in most of the effluents, except for Cyprus and Israel where blaGES was also abundant. Quantitative PCR assessment of blaOXA and blaGES genes in the European effluents revealed similar trends to those displayed in the integron amplicon sequencing pipeline described above, corroborating the robustness of this method and suggesting that these integron-associated genes may be excellent targets for source tracking of effluents in downstream environments. Further application of the above analyses revealed several order-of-magnitude reductions in effluent-associated β-lactamase genes in effluent-saturated soils, suggesting marginal persistence in the soil microbiome.
- Antibiotic resistance genes in treated wastewater and in the receiving water bodies: a pan-European survey of urban settingsPublication . Cacace, Damiano; Fatta-Kassinos, Despo; Manaia, Célia M.; Cytryn, Eddie; Kreuzinger, Norbert; Rizzo, Luigi; Karaolia, Popi; Schwartz, Thomas; Alexander, Johannes; Merlin, Christophe; Garelick, Hemda; Schmitt, Heike; Vries, Daisy de; Schwermer, Carsten U.; Meric, Sureyya; Ozkal, Can Burak; Pons, Marie-Noelle; Kneis, David; Berendonk, Thomas U.There is increasing public concern regarding the fate of antibiotic resistance genes (ARGs) during wastewater treatment, their persistence during the treatment process and their potential impacts on the receiving water bodies. In this study, we used quantitative PCR (qPCR) to determine the abundance of nine ARGs and a class 1 integron associated integrase gene in 16 wastewater treatment plant (WWTP) effluents from ten different European countries. In order to assess the impact on the receiving water bodies, gene abundances in the latter were also analysed. Six out of the nine ARGs analysed were detected in all effluent and river water samples. Among the quantified genes, intI1 and sul1 were the most abundant. Our results demonstrate that European WWTP contribute to the enrichment of the resistome in the receiving water bodies with the particular impact being dependent on the effluent load and local hydrological conditions. The ARGs concentrations in WWTP effluents were found to be inversely correlated to the number of implemented biological treatment steps, indicating a possible option for WWTP management. Furthermore, this study has identified blaOXA-58 as a possible resistance gene for future studies investigating the impact of WWTPs on their receiving water.
- Impact of disinfection processes on bacterial community in urban wastewater: Should we rethink microbial assessment methods?Publication . Cesare, Andrea Di; Corno, Gianluca; Manaia, Célia M.; Rizzo, LuigiThe development of new methods and technologies for microbial characterization as well as their increasing availability at more affordable costs, has made evident the limitations of the conventional and routinely applied (typically cultivation based) methods to exhaustively and fully characterize the actual effect of disinfection process in urban wastewater treatment plants (UWWTPs). Such problem is even more relevant and of concern if microbial challenges, such as the occurrence of pathogens as well as the spread of antibiotic resistance, are taken into account. Such threats move scientists to investigate new and more effective disinfection processes from one side and new methods, techniques and approaches to characterize disinfection process efficiency from the other side. In this opinion paper, the limitations of routine detection methods are discussed according to the relevant and updated scientific literature to explain how research oriented methods and technologies (namely, quantitative real-time PCR, flow cytometry, metagenetics and metagenomics) can allow a better evaluation of disinfection processes. After a short introduction to the main disinfection processes, the application of different microbial characterization methods is discussed according to distinct challenges, such as pathogens inactivation or antibiotic resistance dissemination, when wastewater safety is of concern (for example in reuse scenarios). The routine and research oriented techniques can be successfully used in complementary way to evaluate disinfection process efficiency. Recommendations for UWWTPs managers for internal control of disinfection process are proposed.
- Vancomycin resistant enterococci: from the hospital effluent to the urban wastewater treatment plantPublication . Varela, Ana Rita; Ferro, Giovanna; Vredenburg, Jana; Yanik, Melike; Vieira, Lucas; Rizzo, Luigi; Lameiras, Catarina; Manaia, Célia M.Vancomycin is an important antibiotic to treat serious nosocomial enterococci infections. Human activities, in particular those related with clinical practices performed in hospitals, can potentiate the transfer and selection of clinically-relevant resistant bacteria such as vancomycin resistant enterococci (VRE). Indeed, previous studies demonstrated the occurrence of VRE in urban wastewater treatment plants and related environments (e.g. sewage, rivers). In this study, the occurrence of VRE in a hospital effluent and in the receiving urban wastewater treatment plant was investigated. Vancomycin and ciprofloxacin resistant bacteria occurred in the hospital effluent and in raw municipal inflow at densities of 10(3) to 10(2) CFU mL(-1), being significantly more prevalent in the hospital effluent than in the urban wastewater. Most of the VRE isolated from the hospital effluent belonged to the species Enterococcus faecalis and Enterococcus faecium and presented multidrug-resistance phenotypes to ciprofloxacin, tetracycline, erythromycin, and high-level gentamicin. The same pattern was observed in clinical isolates and in enterococci isolated from the final effluent of the urban wastewater treatment plant. These results show that hospital effluents discharged into urban wastewater treatment plants may be a relevant source of resistance spread to the environment.
- Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a reviewPublication . Rizzo, L.; Manaia, C.; Merlin, C.; Schwartz, T.; Dagot, C.; Ploy, M. C.; Michael, I.; Fatta-Kassinos, D.Urban wastewater treatment plants (UWTPs) are among the main sources of antibiotics' release into the environment. The occurrence of antibiotics may promote the selection of antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB), which shade health risks to humans and animals. In this paper the fate of ARB and ARGs in UWTP's, focusing on different processes/technologies (i.e., biological processes, advanced treatment technologies and disinfection), was critically reviewed. The mechanisms by which biological processes influence the development/selection of ARB and ARGs transfer are still poorly understood. Advanced treatment technologies and disinfection process are regarded as a major tool to control the spread of ARB into the environment. In spite of intense efforts made over the last years to bring solutions to control antibiotic resistance spread in the environment, there are still important gaps to fill in. In particular, it is important to: (i) improve risk assessment studies in order to allow accurate estimates about the maximal abundance of ARB in UWTPs effluents that would not pose risks for human and environmental health; (ii) understand the factors and mechanisms that drive antibiotic resistance maintenance and selection in wastewater habitats. The final objective is to implement wastewater treatment technologies capable of assuring the production of UWTPs effluents with an acceptable level of ARB.
- Immobilised cerium-doped Zinc oxide as a photocatalyst for the degradation of antibiotics and the inactivation of antibiotic-resistant bacteriaPublication . Zammit, Ian; Vaiano, Vincenzo; Ribeiro, Ana R.; Silva, Adrián M. T.; Manaia, Célia M.; Rizzo, LuigiThe threat of antibiotic resistance to the wellbeing of societies is well established. Urban wastewater treatment plants (UWTPs) are recognised sources for antibiotic resistance dissemination in the environment. Herein a novel cerium-doped zinc oxide (Ce-ZnO) photocatalyst is compared to ZnO and the benchmark TiO2-P25 in the immobilised form on a metallic support, to evaluate a photocatalytic process as a possible tertiary treatment in UWTPs. The catalysts were compared for the removal of two antibiotics, trimethoprim (TMP) and sulfamethoxazole (SMX), and for the inactivation of Escherichia coli (E. coli) strain DH5-Alpha in isotonic sodium chloride solution and of autochthonous bacteria in real secondary wastewater. In real wastewater, E. coli and other coliforms were monitored, as well as the respective fractions resistant to ofloxacin and azithromycin. In parallel, Pseudomonas aeruginosa and the respective sub-population resistant to ofloxacin or ciprofloxacin were also monitored. Photocatalysis with both ZnO and Ce-ZnO was faster than using TiO2-P25 at degrading the antibiotics, with Ce-ZnO the fastest against SMX but slower than undoped ZnO in the removal of TMP. Ce-ZnO catalyst reuse in the immobilised form produced somewhat slower kinetics maintained >50% of the initial activity, even after five cycles of use. Approximately 3 log10 inactivation of E. coli in isotonic sodium chloride water was recorded with reproducible results. In the removal of autochthonous bacteria in real wastewater, Ce-ZnO performed better (more than 2 log values higher) than TiO2-P25. In all cases, E. coli and other coliforms, including their resistant subpopulations, were inactivated at a higher rate than P. aeruginosa. With short reaction times no evidence for enrichment of resistance was observed, yet with extended reaction times low levels of bacterial loads were not further inactivated. Overall, Ce-ZnO is an easy and cheap photocatalyst to produce and immobilise and the one that showed higher activity than the industry standard TiO2-P25 against the tested antibiotics and bacteria, including antibiotic-resistant bacteria.
- Wastewater disinfection by solar heterogeneous photocatalysis: effect on tetracycline resistant/sensitive enterococcus strains Enterococcus strainsPublication . Rizzo, L.; Ferro, G.; Manaia, C. M.Solar simulated heterogeneous photocatalysis (SSHP) with suspended TiO2 was investigated in the inactivation of tetracycline resistant/sensitive Enterococcus (TRE/TSE) strains in the effluent of an urban wastewater treatment plant (UWTP). The effect of solar simulated disinfection (SSD) on the inactivation of the same Enterococcus strains was investigated as control. SSHP process (0.05 g l-1 of TiO2) was found to be effective in the inactivation of both Enterococcus strains with total inactivation (7 log unit) observed after 60 min of irradiation. On the contrary, SSD process did not show any significant inactivation after 90 min of irradiation. The effect of both processes on the antibiotic resistance phenotypes of the surviving enterococci was also evaluated. TRE cells surviving the SSHP treatment showed that disinfection process did not affect the antibiotic resistance pattern after 45 min irradiation. The same was observed for the TSE strain. Accordingly, antibiotic resistance can spread into the receiving water body when antibiotic resistant strains survive to disinfection process.