Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms
Objectives: New combinations of β-lactams and β-lactamase inhibitors, such as ceftolozane/tazobactam could be useful to combat biofilm-driven chronic infections by extensively resistant (XDR) Pseudomonas aeruginosa but resistance development by mutations in the Ω-loop of AmpC has been described. How...
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Elsevier
2024-12-01
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| Series: | Biofilm |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S259020752400056X |
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| author | María Fernández-Billón Elena Jordana-Lluch Aina E. Llambías-Cabot María A. Gomis-Font Pablo Fraile-Ribot Rosa I. Torrandell Pamela J. Colman-Vega Óscar Murillo María D. Macià Antonio Oliver |
| author_facet | María Fernández-Billón Elena Jordana-Lluch Aina E. Llambías-Cabot María A. Gomis-Font Pablo Fraile-Ribot Rosa I. Torrandell Pamela J. Colman-Vega Óscar Murillo María D. Macià Antonio Oliver |
| author_sort | María Fernández-Billón |
| collection | DOAJ |
| description | Objectives: New combinations of β-lactams and β-lactamase inhibitors, such as ceftolozane/tazobactam could be useful to combat biofilm-driven chronic infections by extensively resistant (XDR) Pseudomonas aeruginosa but resistance development by mutations in the Ω-loop of AmpC has been described. However, these mutations confer collateral susceptibility to carbapenems. Thus we aimed to evaluate the therapeutic efficacy and the prevention of resistance development of regimen alternating ceftolozane/tazobactam and imipenem. Methods: A carbapenem-resistant XDR P. aeruginosa clinical strain (ST175, 104-B7) and its isogenic imipenem-susceptible ceftolozane/tazobactam-resistant mutant derivative (AmpC T96I, 104-I9) were used. Experiments of single strains and mixed (104-B7 and 104-I9, 1:0.01 ratio) biofilms were performed. 48h biofilms (flow cell system) were treated for 6 days with either ceftolozane/tazobactam, 4/4 mg/L or the alternation of ceftolozane/tazobactam (2 days)-imipenem 4 mg/L (2 days) - ceftolozane/tazobactam (2 days). After treatment, biofilms were collected and plated on Mueller-Hinton agar± ceftolozane/tazobactam 4/4 mg/L. Structural dynamics were monitored using confocal laser scanning microscopy and images were processed with IMARIS software. At least, three independent triplicate experiments per condition were performed. Emerging resistant mutants were characterized through whole genome sequencing (Illumina). Results: Ceftolozane/tazobactam monotherapy failed to reduce the biofilms of the 104-B7 XDR strain and led to the selection of resistant mutants that showed AmpC Ω-loop mutations (T96I, L244R or aa236Δ7). On the contrary, alternation with imipenem enhanced activity (3 Logs reduction at day 6) and prevented the emergence of ceftolozane/tazobactam-resistant mutants. Likewise, treatment with ceftolozane/tazobactam dramatically amplified the resistant strain 104-I9 in mixed biofilms (>90 % of the population), while the alternation regimen counterselected it. Conclusions: Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem effectively prevented the selection of resistant mutants and thus could be a potential therapeutic strategy for the treatment of P. aeruginosa XDR chronic infections. |
| format | Article |
| id | doaj-art-8bebbc899d964ea5b572ef3acde7ec3b |
| institution | DOAJ |
| issn | 2590-2075 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
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| series | Biofilm |
| spelling | doaj-art-8bebbc899d964ea5b572ef3acde7ec3b2025-08-20T02:50:26ZengElsevierBiofilm2590-20752024-12-01810023110.1016/j.bioflm.2024.100231Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilmsMaría Fernández-Billón0Elena Jordana-Lluch1Aina E. Llambías-Cabot2María A. Gomis-Font3Pablo Fraile-Ribot4Rosa I. Torrandell5Pamela J. Colman-Vega6Óscar Murillo7María D. Macià8Antonio Oliver9Department of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, SpainCentro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, Spain; Department of Infectious Diseases, Hospital Universitari de Bellvitge, IDIBELL, Barcelona, SpainDepartment of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, Spain; Corresponding author. Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, Spain.Department of Microbiology, Hospital Universitario Son Espases, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma de Mallorca, Spain; Centro de Investigación Biomédica en Red en Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC), 28029 Madrid, Spain; Corresponding author. Department of Microbiology, Hospital Universitario Son Espases, Ctra. Vallemossa 79, 07120 Palma de Mallorca, Spain.Objectives: New combinations of β-lactams and β-lactamase inhibitors, such as ceftolozane/tazobactam could be useful to combat biofilm-driven chronic infections by extensively resistant (XDR) Pseudomonas aeruginosa but resistance development by mutations in the Ω-loop of AmpC has been described. However, these mutations confer collateral susceptibility to carbapenems. Thus we aimed to evaluate the therapeutic efficacy and the prevention of resistance development of regimen alternating ceftolozane/tazobactam and imipenem. Methods: A carbapenem-resistant XDR P. aeruginosa clinical strain (ST175, 104-B7) and its isogenic imipenem-susceptible ceftolozane/tazobactam-resistant mutant derivative (AmpC T96I, 104-I9) were used. Experiments of single strains and mixed (104-B7 and 104-I9, 1:0.01 ratio) biofilms were performed. 48h biofilms (flow cell system) were treated for 6 days with either ceftolozane/tazobactam, 4/4 mg/L or the alternation of ceftolozane/tazobactam (2 days)-imipenem 4 mg/L (2 days) - ceftolozane/tazobactam (2 days). After treatment, biofilms were collected and plated on Mueller-Hinton agar± ceftolozane/tazobactam 4/4 mg/L. Structural dynamics were monitored using confocal laser scanning microscopy and images were processed with IMARIS software. At least, three independent triplicate experiments per condition were performed. Emerging resistant mutants were characterized through whole genome sequencing (Illumina). Results: Ceftolozane/tazobactam monotherapy failed to reduce the biofilms of the 104-B7 XDR strain and led to the selection of resistant mutants that showed AmpC Ω-loop mutations (T96I, L244R or aa236Δ7). On the contrary, alternation with imipenem enhanced activity (3 Logs reduction at day 6) and prevented the emergence of ceftolozane/tazobactam-resistant mutants. Likewise, treatment with ceftolozane/tazobactam dramatically amplified the resistant strain 104-I9 in mixed biofilms (>90 % of the population), while the alternation regimen counterselected it. Conclusions: Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem effectively prevented the selection of resistant mutants and thus could be a potential therapeutic strategy for the treatment of P. aeruginosa XDR chronic infections.http://www.sciencedirect.com/science/article/pii/S259020752400056XPseudomonas aeruginosaXDRBiofilmsAntagonistic resistance mechanisms to antibiotics |
| spellingShingle | María Fernández-Billón Elena Jordana-Lluch Aina E. Llambías-Cabot María A. Gomis-Font Pablo Fraile-Ribot Rosa I. Torrandell Pamela J. Colman-Vega Óscar Murillo María D. Macià Antonio Oliver Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms Biofilm Pseudomonas aeruginosa XDR Biofilms Antagonistic resistance mechanisms to antibiotics |
| title | Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms |
| title_full | Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms |
| title_fullStr | Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms |
| title_full_unstemmed | Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms |
| title_short | Collateral susceptibility-guided alternation of ceftolozane/tazobactam with imipenem prevents resistance development in XDR Pseudomonas aeruginosa biofilms |
| title_sort | collateral susceptibility guided alternation of ceftolozane tazobactam with imipenem prevents resistance development in xdr pseudomonas aeruginosa biofilms |
| topic | Pseudomonas aeruginosa XDR Biofilms Antagonistic resistance mechanisms to antibiotics |
| url | http://www.sciencedirect.com/science/article/pii/S259020752400056X |
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