Browsing by Author "Purkrtova, Sabina"
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- Biomarkers for monitoring antibiotic resistance in aquatic environmentsPublication . Teixeira, Ana Margarida Ribeiro; Vaz-Moreira, Ivone; Calderón-Franco, David; Weissbrodt, David Gregory; Purkrtova, Sabina; Gajdos, S.; Dottorini, G.; Nielsen, Per Halkjær; Khalifa, Leron; Cytryn, Eddie; Bartacek, Jan; Manaia, CéliaThe occurrence of antimicrobial resistance raises concerns as a human health threat that can be propagated through the environment. Wastewater discharge into the environment is an important source for antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Sewage collection and urban wastewater treatment plants (UWTPs) are major barriers that reduce environmental contamination by sewage-derived pathogens and nutrients. However, the continuous discharge of ARB and ARGs through wastewater, including when well-functioning UWTPs are available, is unavoidable. Regular and integrated antibiotic resistance monitoring in both wastewater and receiving water bodies would contribute to improve control measures. However, monitoring processes are not harmonized being the choice of suitable biomarkers a first limitation. In this study, we tested 10 selected potential antibiotic resistance biomarkers, which have been described has being associated to humans, and rare in clean environments - intI1, sul1, ermB, ermF, aph(3’’)-Ib, qacEΔ1, uidA, mefC, tetX and crAssphage. The public database MGnify (https://www.ebi.ac.uk/metagenomics/; hosted by EMBL-EBI), was screened using the filters corresponding to origin - human gut, wastewater, sewage, and fresh water. These biomarkers and the 16S rRNA gene were monitored by quantitative PCR (qPCR) tested in raw wastewater, activated sludge, treated wastewater and surface water (upstream and downstream the UWTP) samples, collected from different countries (Portugal, Czech Republic, Denmark, The Netherlands, and Israel). The abundance of the 10 potential biomarkers decreased on average by up to 2.5 log-units gene copies/mL of sample from raw wastewater to surface water, due to treatment and/or dilution in surface water. A clustering analysis of samples based on biomarkers abundance, grouped the samples according to the (waste)water type. This classification was confirmed when 12 anonymous (waste)water samples were analysed in a blind test. The tested biomarkers were observed to differentiate different types of sample, permitting the assessment of wastewater treatment efficiency or of impacts of UWTPs discharge or others in aquatic environments. The selection of suitable biomarkers that can typify different water sources and levels of ARG contamination, along with harmonized qPCR procedures, can facilitate regular and integrated legal requirements to antibiotic resistance monitoring in wastewater and related aquatic environments.
- Biomarkers for monitoring antibiotic resistance in aquatic environmentsPublication . Teixeira, A. Margarida; Vaz-Moreira, Ivone; Franco, David Calderón; Weissbrodt, David; Purkrtova, Sabina; Gajdos, Stanislav; Dottorini, Giulia; Nielsen, Per Halkjær; Khalifa, Leron; Cytryn, Eddie; Bartacek, Jan; Manaia, Célia M.
- Candidate biomarkers of antibiotic resistance for the monitoring of wastewater and the downstream environmentPublication . Teixeira, A. Margarida; Vaz-Moreira, Ivone; Calderón-Franco, David; Weissbrodt, David; Purkrtova, Sabina; Gajdos, Stanislav; Dottorini, Giulia; Nielsen, Per Halkjær; Khalifa, Leron; Cytryn, Eddie; Bartacek, Jan; Manaia, Célia M.Urban wastewater treatment plants (UWTPs) are essential for reducing the pollutants load and protecting water bodies. However, wastewater catchment areas and UWTPs emit continuously antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), with recognized impacts on the downstream environments. Recently, the European Commission recommended to monitor antibiotic resistance in UWTPs serving more than 100 000 population equivalents. Antibiotic resistance monitoring in environmental samples can be challenging. The expected complexity of these systems can jeopardize the interpretation capacity regarding, for instance, wastewater treatment efficiency, impacts of environmental contamination, or risks due to human exposure. Simplified monitoring frameworks will be essential for the successful implementation of analytical procedures, data analysis, and data sharing. This study aimed to test a set of biomarkers representative of ARG contamination, selected based on their frequent human association and, simultaneously, rare presence in pristine environments. In addition to the 16S rRNA gene, ten potential biomarkers (intI1, sul1, ermB, ermF, aph(3′’)-Ib, qacEΔ1, uidA, mefC, tetX, and crAssphage) were monitored in DNA extracts (n = 116) from raw wastewater, activated sludge, treated wastewater, and surface water (upstream and downstream of UWTPs) samples collected in the Czech Republic, Denmark, Israel, the Netherlands, and Portugal. Each biomarker was sensitive enough to measure decreases (on average by up to 2.5 log-units gene copy/mL) from raw wastewater to surface water, with variations in the same order of magnitude as for the 16S rRNA gene. The use of the 10 biomarkers allowed the typing of water samples whose origin or quality could be predicted in a blind test. The results show that, based on appropriate biomarkers, qPCR can be used for a cost-effective and technically accessible approach to monitoring wastewater and the downstream environment.
- Monitoring antibiotic resistance genes in wastewater environments: the challenges of filling a gap in the One-Health cyclePublication . Miłobedzka, Aleksandra; Ferreira, Catarina; Vaz-Moreira, Ivone; Calderón-Franco, David; Gorecki, Adrian; Purkrtova, Sabina; Bartacek, Jan; Dziewit, Lukasz; Singleton, Caitlin M.; Nielsen, Per Halkjær; Weissbrodt, David Gregory; Manaia, Célia M.Antibiotic resistance (AR) is a global problem requiring international cooperation and coordinated action. Global monitoring must rely on methods available and comparable across nations to quantify AR occurrence and identify sources and reservoirs, as well as paths of AR dissemination. Numerous analytical tools that are gaining relevance in microbiology, have the potential to be applied to AR research. This review summarizes the state of the art of AR monitoring methods, considering distinct needs, objectives and available resources. Based on the overview of distinct approaches that are used or can be adapted to monitor AR, it is discussed the potential to establish reliable and useful monitoring schemes that can be implemented in distinct contexts. This discussion places the environmental monitoring within the One-Health approach, where two types of risk, dissemination across distinct environmental compartments, and transmission to humans, must be considered. The plethora of methodological approaches to monitor AR and the variable features of the monitored sites challenge the capacity of the scientific community and policy makers to reach a common understanding. However, the dialogue between different methods and the production of action-oriented data is a priority. The review aims to warm up this discussion.
- Monitoring antibiotic resistance in aquatic environmentsPublication . Teixeira, A. Margarida; Cachetas, Diogo; Vaz-Moreira, Ivone; Rocha, Jaqueline; Franco, David Calderón; Weissbrodt, David; Purkrtova, Sabina; Gajdos, Stanislav; Dottorini, Giulia; Nielsen, Per Halkjær; Khalifa, Leron; Cytryn, Eddie; Manaia, Célia M.