Antibiotic resistant and extended-spectrum β-lactamase producing faecal coliforms in wastewater treatment plant effluent

Antibiotic Resistant ◽

Extended Spectrum

AbstractWastewater treatment plants (WWTPs) provide optimal conditions for the maintenance and spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). In this work we describe the occurrence of antibiotic resistant faecal coliforms and their mechanisms of antibiotic resistance in the effluent of two urban WWTPs in Ireland. Effluent samples were collected from two WWTPs in Spring and Autumn of 2015 and 2016. The bacterial susceptibility patterns to 13 antibiotics were determined. The phenotypic tests were carried out to identify AmpC or extended-spectrum β-lactamase (ESBL) producers. The presence of ESBL genes were detected by PCR. Plasmids carrying ESBL genes were transformed into Escherichia coli DH5α recipient and underwent plasmid replicon typing to identify incompatibility groups. More than 90% of isolated faecal coliforms were resistant to amoxicillin and ampicillin, followed by tetracycline (up to 39.82%), ciprofloxacin (up to 31.42%) and trimethoprim (up to 37.61%). Faecal coliforms resistant to colistin and imipenem were detected in all effluent samples. Up to 53.98% of isolated faecal coliforms expressed a multi-drug resistance (MRD) phenotype. AmpC production was confirmed in 5.22% of isolates. The ESBL genes were confirmed for 11.84% of isolates (9.2% of isolates carried blaTEM, 1.4% blaSHV-12, 0.2% blaCTX-M-1 and 1% blaCTX-M-15). Plasmids extracted from 52 ESBL isolates were successfully transformed into recipient E. coli. The detected plasmid incompatibility groups included the IncF group, IncI1, IncHI1/2 and IncA/C. These results provide evidence that treated wastewater is polluted with ARB and MDR faecal coliforms and are sources of ESBL-producing, carbapenem and colistin resistant Enterobacteriaceae.ImportanceAntibiotic resistant bacteria (ARB) are an emerging environmental concern with a potential impact on human health. The results provide the evidence that treated wastewater is polluted with antibiotic resistant bacteria containing mobile resistance mechanisms of importance to clinical treatment of pathogens and multi-drug resistant (MDR) faecal coliforms. They are sources of relatively high proportions of ESBL-producing Enterobacteriaceae, and include carbapenem and colistin resistant Enterobacteriaceae. The significance of this study is the identification of the role of WWTPs as a potential control point to reduce or stop the movement of ESBL, MDR and colistin resistant bacteria into the environment from further upstream sources, such as human or animal waste.

Technologies in Wastewater Treatment Plants for the Removal of Antibiotics, Resistant Bacteria and Antibiotic Resistance Genes: a Review of the Current Literature

Ingenieria y Universidad ◽

10.11144/javeriana.iyu26.twtp ◽

2021 ◽

Vol 26 ◽

Author(s):

Maria Camila Zapata Zúñiga ◽

Miguel Angel Parra-Pérez ◽

Johan Alexander Álvarez-Berrio ◽

Nidia Isabel Molina-Gómez

Keyword(s):

Antibiotic Resistance ◽

Wastewater Treatment ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Resistant Bacteria ◽

Sources Of Information ◽

Antibiotic Resistant ◽

Treatment Processes

This study aimed to evaluate the efficiency of technologies for removing antibiotics, antibiotic-resistant bacteria and their antibiotic resistance genes, and the countries where they have been developed. For this purpose, was conducted a systematic review to identify the tertiary treatments to remove the above-mentioned pollutants. The ScienceDirect and Scopus databases were used as sources of information, taking into account only experimental research from 2006 to 2019 and technologies with removal rates higher than 70% to the information analyses. From the analysis of 9 technologies evaluated, in a set of 47 investigations, photo-Fenton, and electrochemical treatments were found to be the most efficient in the removal of antibiotics; gamma radiation and photocatalysis with TiO2 and UV revealed better results in the removal of resistant microbial agents and their resistance genes, with efficiencies of 99.9%. As one of the largest producers and consumers of antibiotics, China appears to be the country with the most scientific research on the area. The importance of innovation in wastewater treatment processes to achieve better results in the remotion of antibiotics, antibiotic-resistant bacteria, and their resistance genes is highlighted, given the effects on the aquatic ecosystems and public health.

The occurrence of antibiotic-resistant bacteria, including Escherichia coli, in municipal wastewater and river water

E3S Web of Conferences ◽

10.1051/e3sconf/201910000061 ◽

2019 ◽

Vol 100 ◽

pp. 00061 ◽

Cited By ~ 1

Author(s):

Adriana Osińska ◽

Ewa Korzeniewska ◽

Monika Harnisz ◽

Sebastian Niestępski ◽

Piotr Jachimowicz

Keyword(s):

Escherichia Coli ◽

Wastewater Treatment ◽

River Water ◽

Municipal Wastewater ◽

High Efficiency ◽

Treated Wastewater ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli

Wastewater treatment plants (WWTPs) are major reservoirs of antibiotic-resistant bacteria (ARB) which are transported to the natural environment with discharged effluents. Samples of untreated wastewater (UWW) and treated wastewater (TWW) from four municipal WWTPs and samples of river water collected upstream (URW) and downstream (DRW) from the effluent discharge point were analyzed in the study. The total counts of bacteria resistant to β-lactams and tetracyclines and the counts of antibiotic-resistant Escherichia coli were determined. Antibiotic-resistant bacteria, including antibiotic-resistant E. coli, were removed with up to 99.9% efficiency in the evaluated WWTPs. Despite the above, ARB counts in TWW samples were high at up to 1.25x105 CFU/mL in winter and 1.25x103 CFU/mL in summer. Antibiotic-resistant bacteria were also abundant (up to 103 CFU/ml) in URW and DRW samples collected in winter and summer. In both UWW and TWW samples, the counts of ARB and antibiotic-resistant E. coli were at least one order of magnitude lower in summer than in winter. The study revealed that despite the high efficiency of bacterial removal in the wastewater treatment processes, considerable amounts of ARB are released into the environment with TWW and that the percentage of ARB in total bacterial counts increases after wastewater treatment.

Do wastewater treatment plants increase antibiotic resistant bacteria or genes in the environment? Protocol for a systematic review

10.7287/peerj.preprints.27727 ◽

2019 ◽

Author(s):

Daloha Rodríguez-Molina ◽

Petra Mang ◽

Heike Schmitt ◽

Mariana Carmen Chifiriuc ◽

Katja Radon ◽

...

Keyword(s):

Systematic Review ◽

Antibiotic Resistance ◽

Wastewater Treatment ◽

Water Samples ◽

Resistant Bacteria ◽

Process Data ◽

Antibiotic Resistant ◽

Antimicrobial Resistance Genes

Background. Antibiotic resistance is a global public health threat. Water from human activities is collected at wastewater treatment plants where processes often do not sufficiently neutralize antibiotic resistant bacteria and genes, which are further shed into the local environment. This protocol outlines the steps to conduct a systematic review based on the Population, Exposure, Comparator and Outcome (PECO) framework, aiming at answering the question Are antimicrobial-resistant enterobacteriaceae and antimicrobial resistance genes present (O) in air and water samples (P) taken either near or downstream or downwind or down-gradient from wastewater treatment plants (E), as compared to air and water samples taken either further away or upstream or upwind or up-gradient from such wastewater treatment plant (C)? Presence of antimicrobial-resistant bacteria and genes will be quantitatively measured by extracting their prevalence or concentration, depending on the reviewed study. Methods. We will search PubMed, EMBASE, the Cochrane database and Web of Science for original articles published from 01-Jan-2000 to 03-Sep-2018 with language restriction. Articles will undergo a relevance and a design screening process. Data from eligible articles will be extracted by two independent reviewers. Further, we will perform a risk of bias assessment using a decision matrix. We will synthesize and present results in narrative and tabular form and will perform a meta-analysis if heterogeneity of results allows it. Discussion. Antibiotic resistance in environmental samples around wastewater treatment plants may pose a risk of exposure to workers and nearby residents. Results from the systematic review outlined in this protocol will allow to estimate the extend of exposure, to inform policy making and help to design future studies.

Characterization and Quantitation of a Novel β-Lactamase Gene Found in a Wastewater Treatment Facility and the Surrounding Coastal Ecosystem

Applied and Environmental Microbiology ◽

10.1128/aem.02732-10 ◽

2011 ◽

Vol 77 (23) ◽

pp. 8226-8233 ◽

Cited By ~ 32

Author(s):

Miguel I. Uyaguari ◽

Erin B. Fichot ◽

Geoffrey I. Scott ◽

R. Sean Norman

Keyword(s):

Antibiotic Resistance ◽

Wastewater Treatment ◽

Antibiotic Resistance Gene ◽

Significant Proportion ◽

Coastal Ecosystem ◽

Treated Wastewater ◽

Gene Copy ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Content Type

ABSTRACTWastewater treatment plants (WWTPs) are engineered structures that collect, concentrate, and treat human waste, ultimately releasing treated wastewater into local environments. While WWTPs efficiently remove most biosolids, it has been shown that many antibiotics and antibiotic-resistant bacteria can survive the treatment process. To determine how WWTPs influence the concentration and dissemination of antibiotic-resistant genes into the environment, a functional metagenomic approach was used to identify a novel antibiotic resistance gene within a WWTP, and quantitative PCR (qPCR) was used to determine gene copy numbers within the facility and the local coastal ecosystem. From the WWTP metagenomic library, the fosmid insert contained in one highly resistant clone (MIC, ∼416 μg ml−1ampicillin) was sequenced and annotated, revealing 33 putative genes, including a 927-bp gene that is 42% identical to a functionally characterized β-lactamase fromStaphylococcus aureusPC1. Isolation and subcloning of this gene, referred to asblaM-1, conferred ampicillin resistance to itsEscherichia colihost. When normalized to volume, qPCR showed increased concentrations ofblaM-1during initial treatment stages but 2-fold-decreased concentrations during the final treatment stage. The concentration ng−1DNA increased throughout the WWTP process from influent to effluent, suggesting thatblaM-1makes up a significant proportion of the overall genetic material being released into the coastal ecosystem. Average discharge was estimated to be 3.9 × 1014copies of theblaM-1gene released daily into this coastal ecosystem. Furthermore, the gene was observed in all sampled coastal water and sediment samples surrounding the facility. Our results suggest that WWTPs may be a pathway for the dissemination of novel antibiotic resistance genes into the environment.

Characterization of Environmental and Cultivable Antibiotic-Resistant Microbial Communities Associated with Wastewater Treatment

10.20944/preprints202103.0140.v1 ◽

2021 ◽

Author(s):

Alicia Sorgen ◽

James Johnson ◽

Kevin Lambirth ◽

Sandra Clinton ◽

Molly Redmond ◽

...

Keyword(s):

Antibiotic Resistance ◽

Wastewater Treatment ◽

Bacterial Resistance ◽

Compositional Data ◽

Treated Wastewater ◽

Plate Count ◽

Resistant Bacteria ◽

Bacterial Populations ◽

Antibiotic Resistant

Bacterial resistance to antibiotics is a growing global concern, threatening human and environ-mental health, particularly among urban populations. Wastewater treatment plants (WWTPs) are thought to be “hotspots” for antibiotic resistance dissemination. The conditions of WWTPs, in conjunction with the persistence of commonly used antibiotics, may favor the selection and trans-fer of resistance genes among bacterial populations. WWTPs provide an important ecological niche to examine the spread of antibiotic resistance. We used heterotrophic plate count methods to identify phenotypically resistant cultivable portions of these bacterial communities and charac-terized the composition of the culturable subset of these populations. Resistant taxa were more abundant in raw sewage and wastewater before the biological aeration treatment stage. While some antibiotic resistant bacteria (ARB) were detectable downstream of treated wastewater re-lease, these organisms are not enriched relative to effluent-free upstream water, indicating effi-cient removal during treatment. Combined culture-dependent and culture-independent analyses revealed a stark difference in community composition between culturable fractions and the envi-ronmental source material, irrespective of culturing conditions. Higher proportions of the envi-ronmental populations were recovered than predicted by the widely accepted 1% culturability paradigm. These results represent baseline abundance and compositional data for ARB commu-nities for reference in future studies addressing the dissemination of antibiotic resistance associ-ated with urban wastewater treatment ecosystems.

Do wastewater treatment plants increase antibiotic resistant bacteria or genes in the environment? Protocol for a systematic review

10.7287/peerj.preprints.27727v1 ◽

2019 ◽

Author(s):

Daloha Rodríguez-Molina ◽

Petra Mang ◽

Heike Schmitt ◽

Mariana Carmen Chifiriuc ◽

Katja Radon ◽

...

Keyword(s):

Systematic Review ◽

Antibiotic Resistance ◽

Wastewater Treatment ◽

Water Samples ◽

Resistant Bacteria ◽

Process Data ◽

Antibiotic Resistant ◽

Antimicrobial Resistance Genes

Background. Antibiotic resistance is a global public health threat. Water from human activities is collected at wastewater treatment plants where processes often do not sufficiently neutralize antibiotic resistant bacteria and genes, which are further shed into the local environment. This protocol outlines the steps to conduct a systematic review based on the Population, Exposure, Comparator and Outcome (PECO) framework, aiming at answering the question Are antimicrobial-resistant enterobacteriaceae and antimicrobial resistance genes present (O) in air and water samples (P) taken either near or downstream or downwind or down-gradient from wastewater treatment plants (E), as compared to air and water samples taken either further away or upstream or upwind or up-gradient from such wastewater treatment plant (C)? Presence of antimicrobial-resistant bacteria and genes will be quantitatively measured by extracting their prevalence or concentration, depending on the reviewed study. Methods. We will search PubMed, EMBASE, the Cochrane database and Web of Science for original articles published from 01-Jan-2000 to 03-Sep-2018 with language restriction. Articles will undergo a relevance and a design screening process. Data from eligible articles will be extracted by two independent reviewers. Further, we will perform a risk of bias assessment using a decision matrix. We will synthesize and present results in narrative and tabular form and will perform a meta-analysis if heterogeneity of results allows it. Discussion. Antibiotic resistance in environmental samples around wastewater treatment plants may pose a risk of exposure to workers and nearby residents. Results from the systematic review outlined in this protocol will allow to estimate the extend of exposure, to inform policy making and help to design future studies.

Reusing Treated Wastewater: Consideration of the Safety Aspects Associated with Antibiotic-Resistant Bacteria and Antibiotic Resistance Genes

Water ◽

10.3390/w10030244 ◽

2018 ◽

Vol 10 (3) ◽

pp. 244 ◽

Cited By ~ 33

Author(s):

Pei-Ying Hong ◽

Timothy Julian ◽

Marie-Laure Pype ◽

Sunny Jiang ◽

Kara Nelson ◽

...

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Treated Wastewater ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Safety Aspects

A PCR-DGGE approach to evaluate the impact of wastewater source on the antibiotic resistance diversity in treated wastewater effluent

Water Science & Technology ◽

10.2166/wst.2012.024 ◽

2012 ◽

Vol 65 (7) ◽

pp. 1323-1331 ◽

Cited By ~ 4

Author(s):

J. Sigala ◽

A. Unc

Keyword(s):

Antibiotic Resistance ◽

Treatment Plant ◽

Primary Treatment ◽

Treated Wastewater ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Source Type ◽

The Impact

Increased incidence of antibiotics in human-affected environments is raising concerns about increase in acquired antibiotic resistance by environmental bacteria. Wastewater collection and treatment systems are likely significant anthropogenic sinks and vectors for antibiotics and associated antibiotic resistance. Typical municipal treatment plants collect wastewaters of various sources, including well-established antibiotic resistance reservoirs such as hospitals, intensive care units and nursing homes, and integrate them with sources not commonly identified as major sources of antibiotic resistance, such as residential or industrial sources. A comprehensive PCR-DGGE diversity analysis of wastewater antibiotic-resistant bacteria was performed to evaluate the role of various wastewater sources in the discharge of antibiotic resistance by a municipal treatment plant. Wastewater sources are clearly inducing resistance in the final effluent but the role of each source type is highly variable, likely as a function of variable environmental conditions or water use patterns. Comparisons between primary treatment and secondary treatment stages indicate a strong role of the intensity of the wastewater treatment in the diversity profiles of antibiotic-resistant bacteria. While pervasiveness of antibiotic resistance in the system impedes clear discrimination between sources in the tested system, there are indications of specific source type related impacts.

Characterization of Environmental and Cultivable Antibiotic-Resistant Microbial Communities Associated with Wastewater Treatment

Antibiotics ◽

10.3390/antibiotics10040352 ◽

2021 ◽

Vol 10 (4) ◽

pp. 352

Author(s):

Alicia Sorgen ◽

James Johnson ◽

Kevin Lambirth ◽

Sandra M. Clinton ◽

Molly Redmond ◽

...

Keyword(s):

Antibiotic Resistance ◽

Wastewater Treatment ◽

Bacterial Resistance ◽

Compositional Data ◽

Treated Wastewater ◽

Plate Count ◽

Resistant Bacteria ◽

Bacterial Populations ◽

Antibiotic Resistant

Bacterial resistance to antibiotics is a growing global concern, threatening human and environmental health, particularly among urban populations. Wastewater treatment plants (WWTPs) are thought to be “hotspots” for antibiotic resistance dissemination. The conditions of WWTPs, in conjunction with the persistence of commonly used antibiotics, may favor the selection and transfer of resistance genes among bacterial populations. WWTPs provide an important ecological niche to examine the spread of antibiotic resistance. We used heterotrophic plate count methods to identify phenotypically resistant cultivable portions of these bacterial communities and characterized the composition of the culturable subset of these populations. Resistant taxa were more abundant in raw sewage and wastewater before the biological aeration treatment stage. While some antibiotic-resistant bacteria (ARB) were detectable downstream of treated wastewater release, these organisms are not enriched relative to effluent-free upstream water, indicating efficient removal during treatment. Combined culture-dependent and -independent analyses revealed a stark difference in community composition between culturable fractions and the environmental source material, irrespective of culturing conditions. Higher proportions of the environmental populations were recovered than predicted by the widely accepted 1% culturability paradigm. These results represent baseline abundance and compositional data for ARB communities for reference in future studies addressing the dissemination of antibiotic resistance associated with urban wastewater treatment ecosystems.