Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay
Environmental oligotrophic bacteria are suspected to be highly relevant carriers of antimicrobial resistance (AMR). However, there is a lack of validated methods for monitoring in the aquatic environment. Since extended-spectrum β-lactamases (ESBLs) play a particularly important role in the clinical...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-03-01
|
| Series: | Heliyon |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024034157 |
| _version_ | 1797259697121656832 |
|---|---|
| author | Lara Stelmaszyk Claudia Stange Michael Hügler Jatinder P.S. Sidhu Harald Horn Andreas Tiehm |
| author_facet | Lara Stelmaszyk Claudia Stange Michael Hügler Jatinder P.S. Sidhu Harald Horn Andreas Tiehm |
| author_sort | Lara Stelmaszyk |
| collection | DOAJ |
| description | Environmental oligotrophic bacteria are suspected to be highly relevant carriers of antimicrobial resistance (AMR). However, there is a lack of validated methods for monitoring in the aquatic environment. Since extended-spectrum β-lactamases (ESBLs) play a particularly important role in the clinical sector, a culturing method based on R2A-medium spiked with different combinations of β-lactams was applied to quantify β-lactamase-producing environmental bacteria from surface waters. In German surface water samples (n = 28), oligotrophic bacteria ranging from 4.0 × 103 to 1.7 × 104 CFU per 100 mL were detected on the nutrient-poor medium spiked with 3rd generation cephalosporins and carbapenems. These numbers were 3 log10 higher compared to ESBL-producing Enterobacteriales of clinical relevance from the same water samples. A MALDI-TOF MS identification of the isolates demonstrated, that the method leads to the isolation of environmentally relevant strains with Pseudomonas, Flavobacterium, and Janthinobacterium being predominant β-lactam resistant genera. Subsequent micro-dilution antibiotic susceptibility tests (Micronaut-S test) confirmed the expression of β-lactamases. The qPCR analysis of surface waters DNA extracts showed the presence of β-lactamase genes (blaTEM, blaCMY-2, blaOXA-48, blaVIM-2, blaSHV, and blaNDM-1) at concentrations of 3.7 (±1.2) to 1.0 (±1.9) log10 gene copies per 100 mL. Overall, the results demonstrate a widespread distribution of cephalosporinase and carbapenemase enzymes in oligotrophic environmental bacteria that have to be considered as a reservoir of ARGs and contribute to the spread of antibiotic resistance. |
| first_indexed | 2024-04-24T23:13:33Z |
| format | Article |
| id | doaj.art-3f2ebf1bdfef447281e4b9e974bb4a99 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-04-24T23:13:33Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-3f2ebf1bdfef447281e4b9e974bb4a992024-03-17T07:58:02ZengElsevierHeliyon2405-84402024-03-01105e27384Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assayLara Stelmaszyk0Claudia Stange1Michael Hügler2Jatinder P.S. Sidhu3Harald Horn4Andreas Tiehm5TZW: DVGW Technologiezentrum Wasser, Department of Water Microbiology, Karlsruher Straße 84, Karlsruhe, Germany; Corresponding author.TZW: DVGW Technologiezentrum Wasser, Department of Water Microbiology, Karlsruher Straße 84, Karlsruhe, GermanyTZW: DVGW Technologiezentrum Wasser, Department of Water Microbiology, Karlsruher Straße 84, Karlsruhe, GermanyCSIRO Oceans and Atmosphere, Ecosciences Precinct, 41 Boggo Road, Brisbane, AustraliaKarlsruher Institut für Technologie, Engler-Bunte Institute, Wasserchemie und Wassertechnologie, Engler-Bunte-Ring 9a, Karlsruhe, GermanyTZW: DVGW Technologiezentrum Wasser, Department of Water Microbiology, Karlsruher Straße 84, Karlsruhe, GermanyEnvironmental oligotrophic bacteria are suspected to be highly relevant carriers of antimicrobial resistance (AMR). However, there is a lack of validated methods for monitoring in the aquatic environment. Since extended-spectrum β-lactamases (ESBLs) play a particularly important role in the clinical sector, a culturing method based on R2A-medium spiked with different combinations of β-lactams was applied to quantify β-lactamase-producing environmental bacteria from surface waters. In German surface water samples (n = 28), oligotrophic bacteria ranging from 4.0 × 103 to 1.7 × 104 CFU per 100 mL were detected on the nutrient-poor medium spiked with 3rd generation cephalosporins and carbapenems. These numbers were 3 log10 higher compared to ESBL-producing Enterobacteriales of clinical relevance from the same water samples. A MALDI-TOF MS identification of the isolates demonstrated, that the method leads to the isolation of environmentally relevant strains with Pseudomonas, Flavobacterium, and Janthinobacterium being predominant β-lactam resistant genera. Subsequent micro-dilution antibiotic susceptibility tests (Micronaut-S test) confirmed the expression of β-lactamases. The qPCR analysis of surface waters DNA extracts showed the presence of β-lactamase genes (blaTEM, blaCMY-2, blaOXA-48, blaVIM-2, blaSHV, and blaNDM-1) at concentrations of 3.7 (±1.2) to 1.0 (±1.9) log10 gene copies per 100 mL. Overall, the results demonstrate a widespread distribution of cephalosporinase and carbapenemase enzymes in oligotrophic environmental bacteria that have to be considered as a reservoir of ARGs and contribute to the spread of antibiotic resistance.http://www.sciencedirect.com/science/article/pii/S2405844024034157Antibiotic resistanceEnvironmental bacteriaCulture-based methodsMALDI-TOF MSqPCRβ-lactamases |
| spellingShingle | Lara Stelmaszyk Claudia Stange Michael Hügler Jatinder P.S. Sidhu Harald Horn Andreas Tiehm Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay Heliyon Antibiotic resistance Environmental bacteria Culture-based methods MALDI-TOF MS qPCR β-lactamases |
| title | Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay |
| title_full | Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay |
| title_fullStr | Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay |
| title_full_unstemmed | Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay |
| title_short | Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay |
| title_sort | quantification of β lactamase producing bacteria in german surface waters with subsequent maldi tof ms based identification and β lactamase activity assay |
| topic | Antibiotic resistance Environmental bacteria Culture-based methods MALDI-TOF MS qPCR β-lactamases |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024034157 |
| work_keys_str_mv | AT larastelmaszyk quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay AT claudiastange quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay AT michaelhugler quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay AT jatinderpssidhu quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay AT haraldhorn quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay AT andreastiehm quantificationofblactamaseproducingbacteriaingermansurfacewaterswithsubsequentmalditofmsbasedidentificationandblactamaseactivityassay |