An assay for determining minimal concentrations of antibiotics that drive horizontal transfer of resistance
| Autor: | Jekaterina Jutkina, Carolin Rutgersson, D. G. Joakim Larsson, Carl-Fredrik Flach |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Environmental Engineering Gene Transfer Horizontal Tetracycline medicine.drug_class 030106 microbiology Antibiotics Microbial Sensitivity Tests 010501 environmental sciences Biology medicine.disease_cause 01 natural sciences Microbiology 03 medical and health sciences Minimum inhibitory concentration Antibiotic resistance Plasmid medicine Environmental Chemistry Waste Management and Disposal Escherichia coli 0105 earth and related environmental sciences Drug Resistance Microbial biology.organism_classification Pollution Anti-Bacterial Agents Horizontal gene transfer Biological Assay Bacteria medicine.drug |
| Zdroj: | The Science of the total environment. |
| ISSN: | 1879-1026 |
| Popis: | Ability to understand the factors driving horizontal transfer of antibiotic resistance from unknown, harmless bacteria to pathogens is crucial in order to tackle the growing resistance problem. However, current methods to measure effects of stressors on horizontal gene transfer have limitations and often fall short, as the estimated endpoints can be a mix of both the number of transfer events and clonal growth of transconjugants. Our aim was therefore to achieve a proper strategy for assessing the minimal concentration of a stressor (exemplified by tetracycline) that drives horizontal transfer of antibiotic resistance from a complex community to a model pathogen. Conditions were optimized to improve a culture-based approach using the bacterial community of treated sewage effluent as donor, and fluorescent, traceable Escherichia coli as recipient. Reduced level of background resistance, differentiation of isolates as well as decreased risk for measuring effects of selection were achieved through the use of chromogenic medium, optimization of conjugation time as well as applying a different antibiotic for isolation of transconjugants than the one tested for its ability to drive transfer. Using this assay, we showed that a very low concentration of tetracycline, 10μg/L i.e. 150 times below the minimal inhibitory concentration of the recipient, promoted horizontal transfer of multiple antibiotic-resistance determinants. Higher concentrations favoured selection of a tetracycline-resistance phenotype along with a decline in the number of detectable transfer events. The described method can be used to evaluate different environmental conditions and factors that trigger horizontal dissemination of mobile resistance elements, eventually resulting in the formation of drug-resistant pathogens. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect