Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach
Determining the prevalence of antimicrobial resistance (AMR) in non-clinical settings is vital for better management of the global AMR crisis. Untreated and even treated wastewaters are important sources that release AMR into the environment. Methodologically, it is difficult to generate a comprehen...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-06-01
|
| Series: | Water |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4441/15/13/2432 |
| _version_ | 1797590778661306368 |
|---|---|
| author | Camila A. Knecht Maja Hinkel Ines Mäusezahl Anne-Kristin Kaster Jaime Nivala Jochen A. Müller |
| author_facet | Camila A. Knecht Maja Hinkel Ines Mäusezahl Anne-Kristin Kaster Jaime Nivala Jochen A. Müller |
| author_sort | Camila A. Knecht |
| collection | DOAJ |
| description | Determining the prevalence of antimicrobial resistance (AMR) in non-clinical settings is vital for better management of the global AMR crisis. Untreated and even treated wastewaters are important sources that release AMR into the environment. Methodologically, it is difficult to generate a comprehensive in situ profile of antibiotic resistance gene hosts. Here, we used epicPCR (emulsion, paired isolation, and concatenation PCR) as a cultivation-independent method to reveal the host profiles of the AMR indicator genes <i>intI1</i>, <i>sul1</i>, <i>sul2</i>, and <i>dfrA1</i> in two constructed wetlands treating municipal wastewater. Overall, the epicPCR analysis revealed a profile of AMR indicator gene hosts that is consistent with literature data from cultivation-based approaches. Most carriers of antibiotic resistance (AR) genes and likely of class 1 integrons belonged to the <i>Gammaproteobateria</i>, particularly the <i>Burkholderiaceae</i> and <i>Rhodocyclaceae</i> families, followed by members of the <i>Campylobacterota</i>, <i>Desulfobacterota</i>, and <i>Firmicutes</i>. The analysis also identified several novel hosts for the indicator genes widely distributed in the wetlands, including the genera <i>Legionella</i> and <i>Ralstonia</i>. Therefore, the application of epicPCR has produced an expanded insight into the in situ indicator gene host profile, while highlighting the role of the environment as a reservoir for AMR. |
| first_indexed | 2024-03-11T01:25:21Z |
| format | Article |
| id | doaj.art-4bbec56267d24c1695257287f6b00867 |
| institution | Directory Open Access Journal |
| issn | 2073-4441 |
| language | English |
| last_indexed | 2024-03-11T01:25:21Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Water |
| spelling | doaj.art-4bbec56267d24c1695257287f6b008672023-11-18T17:48:09ZengMDPI AGWater2073-44412023-06-011513243210.3390/w15132432Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput ApproachCamila A. Knecht0Maja Hinkel1Ines Mäusezahl2Anne-Kristin Kaster3Jaime Nivala4Jochen A. Müller5Department of Environmental Biotechnology, Helmholtz Centre for Environmental Research—UFZ, 04318 Leipzig, GermanyDepartment of Environmental Biotechnology, Helmholtz Centre for Environmental Research—UFZ, 04318 Leipzig, GermanyDepartment of Environmental Biotechnology, Helmholtz Centre for Environmental Research—UFZ, 04318 Leipzig, GermanyInstitute for Biological Interfaces (IBG-5), Karlsruhe Institute of Technology, 76344 Eggenstein-Leopoldshafen, GermanyEnvironmental and Biotechnology Centre, Helmholtz Centre for Environmental Research—UFZ, 04318 Leipzig, GermanyDepartment of Environmental Biotechnology, Helmholtz Centre for Environmental Research—UFZ, 04318 Leipzig, GermanyDetermining the prevalence of antimicrobial resistance (AMR) in non-clinical settings is vital for better management of the global AMR crisis. Untreated and even treated wastewaters are important sources that release AMR into the environment. Methodologically, it is difficult to generate a comprehensive in situ profile of antibiotic resistance gene hosts. Here, we used epicPCR (emulsion, paired isolation, and concatenation PCR) as a cultivation-independent method to reveal the host profiles of the AMR indicator genes <i>intI1</i>, <i>sul1</i>, <i>sul2</i>, and <i>dfrA1</i> in two constructed wetlands treating municipal wastewater. Overall, the epicPCR analysis revealed a profile of AMR indicator gene hosts that is consistent with literature data from cultivation-based approaches. Most carriers of antibiotic resistance (AR) genes and likely of class 1 integrons belonged to the <i>Gammaproteobateria</i>, particularly the <i>Burkholderiaceae</i> and <i>Rhodocyclaceae</i> families, followed by members of the <i>Campylobacterota</i>, <i>Desulfobacterota</i>, and <i>Firmicutes</i>. The analysis also identified several novel hosts for the indicator genes widely distributed in the wetlands, including the genera <i>Legionella</i> and <i>Ralstonia</i>. Therefore, the application of epicPCR has produced an expanded insight into the in situ indicator gene host profile, while highlighting the role of the environment as a reservoir for AMR.https://www.mdpi.com/2073-4441/15/13/2432antimicrobial resistancewastewaterconstructed wetlandsingle-cell analysisepicPCR |
| spellingShingle | Camila A. Knecht Maja Hinkel Ines Mäusezahl Anne-Kristin Kaster Jaime Nivala Jochen A. Müller Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach Water antimicrobial resistance wastewater constructed wetland single-cell analysis epicPCR |
| title | Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach |
| title_full | Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach |
| title_fullStr | Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach |
| title_full_unstemmed | Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach |
| title_short | Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach |
| title_sort | identification of antibiotic resistance gene hosts in treatment wetlands using a single cell based high throughput approach |
| topic | antimicrobial resistance wastewater constructed wetland single-cell analysis epicPCR |
| url | https://www.mdpi.com/2073-4441/15/13/2432 |
| work_keys_str_mv | AT camilaaknecht identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach AT majahinkel identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach AT inesmausezahl identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach AT annekristinkaster identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach AT jaimenivala identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach AT jochenamuller identificationofantibioticresistancegenehostsintreatmentwetlandsusingasinglecellbasedhighthroughputapproach |