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Real-Time Monitorin...
Real-Time Monitoring of nfxB Mutant Occurrence and Dynamics in Pseudomonas aeruginosa Biofilm Exposed to Subinhibitory Concentrations of Ciprofloxacin
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- Zaborskyte, Greta (författare)
- Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi,Univ Copenhagen, Dept Immunol & Microbiol, Costerton Biofilm Ctr, Copenhagen, Denmark.
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- Andersen, Jens Bo (författare)
- Univ Copenhagen, Dept Immunol & Microbiol, Costerton Biofilm Ctr, Copenhagen, Denmark.
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- Kragh, Kasper Norskov (författare)
- Univ Copenhagen, Dept Immunol & Microbiol, Costerton Biofilm Ctr, Copenhagen, Denmark.
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- Ciofu, Oana (författare)
- Univ Copenhagen, Dept Immunol & Microbiol, Costerton Biofilm Ctr, Copenhagen, Denmark.
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(creator_code:org_t)
- AMER SOC MICROBIOLOGY, 2017
- 2017
- Engelska.
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Ingår i: Antimicrobial Agents and Chemotherapy. - : AMER SOC MICROBIOLOGY. - 0066-4804 .- 1098-6596. ; 61:3
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Biofilm infections caused by Pseudomonas aeruginosa are frequently treated with ciprofloxacin (CIP); however, resistance rapidly develops. One of the primary resistance mechanisms is the overexpression of the MexCD-OprJ pump due to a mutation in nfxB, encoding the transcriptional repressor of this pump. The aim of this study was to investigate the effect of subinhibitory concentrations of CIP on the occurrence of nfxB mutants in the wild-type PAO1 flow cell biofilm model. For this purpose, we constructed fluorescent reporter strains (PAO1 background) with an mCherry tag for constitutive red fluorescence and chromosomal transcriptional fusion between the P-mexCD promoter and gfp leading to green fluorescence upon mutation of nfxB. We observed a rapid development of nfxB mutants by live confocal laser scanning microscopy (CLSM) imaging of the flow cell biofilm (reaching 80 to 90% of the whole population) when treated with 1/10 minimal biofilm inhibitory concentration of CIP for 24 h and 96 h. Based on the observed developmental stages, we propose that nfxB mutants emerged de novo in the biofilm during CIP treatment from filamentous cells, which might have arisen due to the stress responses induced by CIP. Identical nfxB mutations were found in fluorescent colonies from the same flow cell biofilm, especially in 24-h biofilms, suggesting selection and clonal expansion of the mutants during biofilm growth. Our findings point at the significant role of high-enough antibiotic dosages or appropriate combination therapy to avoid the emergence of resistant mutants in biofilms.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Microbiology in the medical area (hsv//eng)
Nyckelord
- Pseudomonas aeruginosa
- biofilms
- ciprofloxacin
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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