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Development of in vitro resistance to fluoroquinolones in Pseudomonas aeruginosa
Xiaojing He
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9586-6250
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This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Antimicrobial Resistance and Infection Control.
Fluoroquinolone resistance in Pseudomonas aeruginosa typically arises through site-specific mutations and overexpression of efflux pumps. In this study, we investigated the dynamics of different resistance mechanisms in P. aeruginosa populations that have evolved under fluoroquinolone pressure, as well as the interactions between these mechanisms in evolutionary trajectories. Bacteria of strain ATCC27853 were selected under different concentrations of ciprofloxacin and levofloxacin for six parallel lineages, followed by amplification of four target genes in the quinolone-resistance determining region (QRDR) and Sanger sequencing to identify the mutations. The expression of four efflux pump proteins was evaluated by real-time polymerase chain reaction using the relative quantitation method, with the ATCC27853 strain used as a control. We found that ciprofloxacin killed P. aeruginosa sooner than did levofloxacin. Further, we identified five different mutations in three subunits of QRDRs, with gyrA as the main mutated gene associated with conferring fluoroquinolone resistance. Additionally, we found a larger number of mutations appearing at 2 mg/L and 4 mg/L of ciprofloxacin and levofloxacin, respectively. Moreover, we identified the main efflux pump being expressed as MexCD-OprJ, with initial overexpression observed at 0.25 mg/L and 0.5 mg/L of ciprofloxacin and levofloxacin, respectively. These results demonstrated gyrA83 mutation and MexCD-OprJ overexpression as the primary mechanism conferring ciprofloxacin and levofloxacin resistance in P. aeruginosa. In addition, we also show that ciprofloxacin exhibited a stronger ability to kill the bacteria while potentially rendering it more susceptible to resistance.
Keywords
Fluoroquinolone, Pseudomonas aeruginosa, Efflux pumps, Levofloxacin, Ciprofloxacin
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CURRENT STATUS: Published
Version 2
Posted 23 Jun, 2020
Published
On 05 Aug, 2020
No community comments so far
Editorial decision: Accept
On 16 Jul, 2020
Reviewer #2 agreed
On 01 Jul, 2020
Review #2 received
Received 01 Jul, 2020
Review #1 received
Received 28 Jun, 2020
Reviewers invited
Invitations sent on 26 Jun, 2020
Reviewer #1 agreed
On 26 Jun, 2020
Editor assigned
On 23 Jun, 2020
Submission checks complete
On 22 Jun, 2020
Editor invited
On 22 Jun, 2020
No comments provided
Editorial decision: Major revision
On 02 Jun, 2020
Review #1 received
Received 09 May, 2020
Review #2 received
Received 03 May, 2020
Reviewer #2 agreed
On 21 Apr, 2020
Reviewer #1 agreed
On 17 Apr, 2020
Reviewers invited
Invitations sent on 16 Apr, 2020
Editor assigned
On 15 Apr, 2020
Editor invited
On 14 Apr, 2020
Submission checks complete
On 06 Apr, 2020
First submitted
On 31 Mar, 2020
Metrics
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This preprint is under consideration at Antimicrobial Resistance and Infection Control. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Development of in vitro resistance to fluoroquinolones in Pseudomonas aeruginosa
Xiaojing He
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9586-6250
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 2
Posted 23 Jun, 2020
Published
On 05 Aug, 2020
No community comments so far
Editorial decision: Accept
On 16 Jul, 2020
Reviewer #2 agreed
On 01 Jul, 2020
Review #2 received
Received 01 Jul, 2020
Review #1 received
Received 28 Jun, 2020
Reviewers invited
Invitations sent on 26 Jun, 2020
Reviewer #1 agreed
On 26 Jun, 2020
Editor assigned
On 23 Jun, 2020
Submission checks complete
On 22 Jun, 2020
Editor invited
On 22 Jun, 2020
No comments provided
Editorial decision: Major revision
On 02 Jun, 2020
Review #1 received
Received 09 May, 2020
Review #2 received
Received 03 May, 2020
Reviewer #2 agreed
On 21 Apr, 2020
Reviewer #1 agreed
On 17 Apr, 2020
Reviewers invited
Invitations sent on 16 Apr, 2020
Editor assigned
On 15 Apr, 2020
Editor invited
On 14 Apr, 2020
Submission checks complete
On 06 Apr, 2020
First submitted
On 31 Mar, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Antimicrobial Resistance and Infection Control.
Fluoroquinolone resistance in Pseudomonas aeruginosa typically arises through site-specific mutations and overexpression of efflux pumps. In this study, we investigated the dynamics of different resistance mechanisms in P. aeruginosa populations that have evolved under fluoroquinolone pressure, as well as the interactions between these mechanisms in evolutionary trajectories. Bacteria of strain ATCC27853 were selected under different concentrations of ciprofloxacin and levofloxacin for six parallel lineages, followed by amplification of four target genes in the quinolone-resistance determining region (QRDR) and Sanger sequencing to identify the mutations. The expression of four efflux pump proteins was evaluated by real-time polymerase chain reaction using the relative quantitation method, with the ATCC27853 strain used as a control. We found that ciprofloxacin killed P. aeruginosa sooner than did levofloxacin. Further, we identified five different mutations in three subunits of QRDRs, with gyrA as the main mutated gene associated with conferring fluoroquinolone resistance. Additionally, we found a larger number of mutations appearing at 2 mg/L and 4 mg/L of ciprofloxacin and levofloxacin, respectively. Moreover, we identified the main efflux pump being expressed as MexCD-OprJ, with initial overexpression observed at 0.25 mg/L and 0.5 mg/L of ciprofloxacin and levofloxacin, respectively. These results demonstrated gyrA83 mutation and MexCD-OprJ overexpression as the primary mechanism conferring ciprofloxacin and levofloxacin resistance in P. aeruginosa. In addition, we also show that ciprofloxacin exhibited a stronger ability to kill the bacteria while potentially rendering it more susceptible to resistance.
Figure 1
Figure 2
Figure 3