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Investigating the impact of UV-C/H2O2 and sunlight/H2O2 on the removal of antibiotics, antibiotic resistance determinants and toxicity present in urban wastewater

Publication . G. Michael, Stella; Michael-Kordatou, Irene; Nahim-Granados, Samira; Polo-López, Maria Inmaculada; Rocha, Jaqueline; Martínez-Piernas, Ana B.; Fatta-Kassinos, Pilar; Manaia, Célia M.

This work aimed at exploring the impact of UV-C/H2O2 and sunlight/H2O2 processes, applied at pilot scale, on removing: (i) ciprofloxacin and sulfamethoxazole, (ii) cultivable Escherichia coli and Pseudomonas aeruginosa grown in the presence and absence of sub-minimal inhibitory concentrations of ciprofloxacin and sulfamethoxazole and (iii) the genes 16S rRNA and selected antibiotic resistance genes (ARGs) (i.e., sul1, blaCTX-M, qnrS, tetM, etc.) from urban wastewater. The major antibiotic transformation products (TPs) formed, were elucidated and the chronic toxicity of the whole effluent mixture against Vibrio fischeri was evaluated. The capability of the processes, in terms of the elimination of the antibiotics present in urban wastewater, varied among the two light sources used: both antibiotics were fully removed during UV-C/Η2Ο2, whereas only ciprofloxacin was removed during the sunlight/H2O2. The photo-transformation of the antibiotics led to the identification of 21 and 18 TPs of ciprofloxacin and sulfamethoxazole, respectively, while all of them retained their core moiety, responsible for the antibacterial activity. All the UV-C/H2O2-treated samples were found to be toxic, whereas the luminescence of V. fischeri was not inhibited when tested in the sunlight/H2O2-treated samples. During both processes, E. coli, P. aeruginosa and the colonies of these species still viable in the presence of antibiotics, were successfully inactivated to values below the detection limit. However, sunlight/H2O2 has not achieved complete disinfection, as regrowth of E. coli and P. aeruginosa colonies was observed after 48 h of storage of the treated effluent. Finally, none of the technologies tested was able to completely remove the target ARGs, confirming their inability to prevent the spread of resistance determinants to the environment.

2020Journal article

Restricted access

Betaproteobacteria are predominant in drinking water: are there reasons for concern?

Publication . Ferro, Pompeyo; Vaz-Moreira, Ivone; Manaia, Célia M.

Betaproteobacteria include some of the most abundant and ubiquitous bacterial genera that can be found in drinking water, including mineral water. The combination of physiology and ecology traits place some Betaproteobacteria in the list of potential, yet sometimes neglected, opportunistic pathogens that can be transmitted by water or aqueous solutions. Indeed, some drinking water Betaproteobacteria with intrinsic and sometimes acquired antibiotic resistance, harbouring virulence factors and often found in biofilm structures, can persist after water disinfection and reach the consumer. This literature review summarises and discusses the current knowledge about the occurrence and implications of Betaproteobacteria in drinking water. Although the sparse knowledge on the ecology and physiology of Betaproteobacteria thriving in tap or bottled natural mineral/spring drinking water (DW) is an evidence of this review, it is demonstrated that DW holds a high diversity of Betaproteobacteria, whose presence may not be innocuous. Frequently belonging to genera also found in humans, DW Betaproteobacteria are ubiquitous in different habitats, have the potential to resist antibiotics either due to intrinsic or acquired mechanisms, and hold different virulence factors. The combination of these factors places DW Betaproteobacteria in the list of candidates of emerging opportunistic pathogens. Improved bacterial identification of clinical isolates associated with opportunistic infections and additional genomic and physiological studies may contribute to elucidate the potential impact of these bacteria.

2019Journal article

Open access

Neighbor urban wastewater treatment plants display distinct profiles of bacterial community and antibiotic resistance genes

Publication . Fernandes, Telma; Vaz-Moreira, Ivone; Manaia, Célia M.

Urban wastewater treatment plants (UWTPs) are among the major recipients of antibiotic-resistant bacteria (ARB), antibiotic resistance genes (ARGs), and antibiotic residues in urban environments. Although during treatment, bacteria of human and animal origin are removed, some are able to survive, persisting in the final effluent. The occurrence of these bacteria, especially those harboring ARGs, may have a direct impact on the quality of the treated wastewater that is returned to the environment. In this study, we aimed to assess if the final effluent bacterial communities of three UWTPs (PT1, PT2, and PT3) located next to each other were distinct and if such differences were related with the antibiotic resistance profiles. It was observed that the bacterial community (16S rRNA gene Illumina sequencing) and load of selected ARGs of final effluent differed among the three UWTPs, irrespective of sampling time. Members of the families Aeromonadaceae, Campylobacteraceae, Veillonellaceae, [Weeksellaceae], and Porphyromonadaceae were observed to be positively correlated with some ARGs (bla(CTX-M), bla(OXA-A), bla(SHV)) and intI1 (p < 0.05), while Intrasporangiaceae were observed to be negatively correlated. While Aeromonadaceae are recognized relevant ARG harbors, the other bacterial families may represent bacteria that co-exist with the ARG hosts, which may belong to minor bacterial groups omitted in the analyses. These findings suggest the importance of bacterial dynamics during treatment to the ARB&ARGs removal, a rationale that may contribute to design new strategies to apply in the UWTPs to prevent the spread of antibiotic resistance.

2019Journal article

Open access

A rationale for the high limits of quantification of antibiotic resistance genes in soil

Publication . Fortunato, Gianuario; Vaz-Moreira, Ivone; Becerra-Castro, Cristina; Nunes, Olga C.; Manaia, Célia M.

The determination of values of abundance of antibiotic resistance genes (ARGs) per mass of soil is extremely useful to assess the potential impacts of relevant sources of antibiotic resistance, such as irrigation with treated wastewater or manure application. Culture-independent methods and, in particular, quantitative PCR (qPCR), have been regarded as suitable approaches for such a purpose. However, it is arguable if these methods are sensitive enough to measure ARGs abundance at levels that may represent a risk for environmental and human health. This study aimed at demonstrating the range of values of ARGs quantification that can be expected based on currently used procedures of DNA extraction and qPCR analyses. The demonstration was based on the use of soil samples spiked with known amounts of wastewater antibiotic resistant bacteria (ARB) (Enterococcus faecalis, Escherichia coli, Acinetobacter johnsonii, or Pseudomonas aeruginosa), harbouring known ARGs, and also on the calculation of expected values determined based on qPCR. The limits of quantification (LOQ) of the ARGs (vanA, qnrS, blaTEM, blaOXA, blaIMP, blaVIM) were observed to be approximately 4 log-units per gram of soil dry weight, irrespective of the type of soil tested. These values were close to the theoretical LOQ values calculated based on currently used DNA extraction methods and qPCR procedures. The observed LOQ values can be considered extremely high to perform an accurate assessment of the impacts of ARGs discharges in soils. A key message is that ARGs accumulation will be noticeable only at very high doses. The assessment of the impacts of ARGs discharges in soils, of associated risks of propagation and potential transmission to humans, must take into consideration this type of evidence, and avoid the simplistic assumption that no detection corresponds to risk absence.

2018Journal article

Open access

Hydromonas duriensis gen. nov., sp. nov., isolated from freshwater

Publication . Vaz-Moreira, Ivone; Narciso-da-Rocha, Carlos; De Brandt, Evie; Vandamme, Peter; Ferreira, A. C. Silva; Lobo-da-Cunha, Alexandre; Nunes, Olga C.; Manaia, Célia M.

An aerobic, Gram-stain-negative rod, designated strain A2P5T , was isolated from the Douro river, in Porto, Portugal. Cells were catalase- and oxidase-positive. Growth occurred at15–30 8C, at pH 6–8 and in the presence of 1 % (w/v) NaCl. The major respiratory quinone was Q8, the genomic DNA had a G+C content of 47¡1 mol%, and phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol were amongst the major polar lipids. On the basis of 16S rRNA gene sequence analysis, strain A2P5T was observed to be a member of the family Burkholderiaceae, but could not be identified as a member of any validly named genus. The low levels of 16S rRNA gene sequence similarity to other recognized taxa (,91 %), together with the comparative analysis of phenotypic and chemotaxonomic characteristics, supported the proposal of a novel species of a new genus within the family Burkholderiaceae. The name Hydromonas duriensis gen. nov., sp. nov. is proposed. The type strain of Hydromonas duriensis is A2P5T (5LMG 28428T 5CCUG 66137T).

2015Journal article

Restricted access