Increased bactericidal activity of colistin on Pseudomonas aeruginosa biofilms in anaerobic conditions

Tolerance towards antibiotics of Pseudomonas aeruginosa biofilms is recognized as a major cause of therapeutic failure of chronic lung infection in cystic fibrosis (CF) patients. This lung infection is characterized by antibiotic-tolerant biofilms in mucus with zones of O2 depletion mainly due to polymorphonuclear leukocytic activity. In contrast to the main types of bactericidal antibiotics, it has not been possible to establish an association between the bactericidal effects of colistin and the production of detectable levels of OH ˙ on several strains of planktonic P. aeruginosa. Therefore, we propose that production of OH ˙ may not contribute significantly to the bactericidal activity of colistin on P. aeruginosa biofilm. Thus, we investigated the effect of colistin treatment on biofilm of wild-type PAO1, a catalase-deficient mutant (ΔkatA) and a colistin-resistant CF isolate cultured in microtiter plates in normoxic- or anoxic atmosphere with 1 mM nitrate. The killing of bacteria during colistin treatment was measured by CFU counts, and the OH⋅ formation was measured by 3′-(p-hydroxylphenyl fluorescein) fluorescein (HPF) fluorescence. Validation of the assay was done by hydrogen peroxide treatment. OH⋅ formation was undetectable in aerobic PAO1 biofilms during 3 h of colistin treatment. Interestingly, we demonstrate increased susceptibility of P. aeruginosa biofilms towards colistin during anaerobic conditions. In fact, the maximum enhancement of killing by anaerobic conditions exceeded 2 logs using 4 mg L−1 of colistin compared to killing at aerobic conditions.

[1]  G. Di Bonaventura,et al.  In vitro activity of colistin against biofilm by Pseudomonas aeruginosa is significantly improved under "cystic fibrosis-like" physicochemical conditions. , 2015, Diagnostic microbiology and infectious disease.

[2]  M. Gladwin,et al.  Sodium Nitrite Blocks the Activity of Aminoglycosides against Pseudomonas aeruginosa Biofilms , 2015, Antimicrobial Agents and Chemotherapy.

[3]  M. Kühl,et al.  Physiological levels of nitrate support anoxic growth by denitrification of Pseudomonas aeruginosa at growth rates reported in cystic fibrosis lungs and sputum , 2014, Front. Microbiol..

[4]  T. Scheike,et al.  Polymorphonuclear Leukocytes Restrict Growth of Pseudomonas aeruginosa in the Lungs of Cystic Fibrosis Patients , 2014, Infection and Immunity.

[5]  C. Hansen,et al.  Nitric oxide production by polymorphonuclear leucocytes in infected cystic fibrosis sputum consumes oxygen , 2014, Clinical and experimental immunology.

[6]  Ahmad S Khalil,et al.  Antibiotics induce redox-related physiological alterations as part of their lethality. , 2014, Proceedings of the National Academy of Sciences of the United States of America.

[7]  N. Høiby,et al.  Formation of hydroxyl radicals contributes to the bactericidal activity of ciprofloxacin against Pseudomonas aeruginosa biofilms. , 2014, Pathogens and disease.

[8]  N. Høiby,et al.  Bactericidal effect of colistin on planktonic Pseudomonas aeruginosa is independent of hydroxyl radical formation. , 2014, International journal of antimicrobial agents.

[9]  A. Kharazmi,et al.  Nitrous Oxide Production in Sputum from Cystic Fibrosis Patients with Chronic Pseudomonas aeruginosa Lung Infection , 2014, PloS one.

[10]  H. Nelis,et al.  Biofilm-Grown Burkholderia cepacia Complex Cells Survive Antibiotic Treatment by Avoiding Production of Reactive Oxygen Species , 2013, PloS one.

[11]  J. Imlay,et al.  Cell Death from Antibiotics Without the Involvement of Reactive Oxygen Species , 2013, Science.

[12]  R. Kaul,et al.  Polymyxin Resistance of Pseudomonas aeruginosa phoQ Mutants Is Dependent on Additional Two-Component Regulatory Systems , 2013, Antimicrobial Agents and Chemotherapy.

[13]  M. Strous,et al.  Denitrification and aerobic respiration, hybrid electron transport chains and co-evolution. , 2013, Biochimica et biophysica acta.

[14]  T. Tolker-Nielsen,et al.  The metabolically active subpopulation in Pseudomonas aeruginosa biofilms survives exposure to membrane-targeting antimicrobials via distinct molecular mechanisms. , 2012, FEMS immunology and medical microbiology.

[15]  Samuel I. Miller,et al.  PmrB Mutations Promote Polymyxin Resistance of Pseudomonas aeruginosa Isolated from Colistin-Treated Cystic Fibrosis Patients , 2011, Antimicrobial Agents and Chemotherapy.

[16]  Samuel I. Miller,et al.  PhoQ Mutations Promote Lipid A Modification and Polymyxin Resistance of Pseudomonas aeruginosa Found in Colistin-Treated Cystic Fibrosis Patients , 2011, Antimicrobial Agents and Chemotherapy.

[17]  N. Høiby,et al.  Pharmacokinetics/Pharmacodynamics of Colistin and Imipenem on Mucoid and Nonmucoid Pseudomonas aeruginosa Biofilms , 2011, Antimicrobial Agents and Chemotherapy.

[18]  C. von Buchwald,et al.  Decreased mucosal oxygen tension in the maxillary sinuses in patients with cystic fibrosis. , 2011, Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society.

[19]  N. Høiby,et al.  Pseudomonas aeruginosa biofilms in cystic fibrosis. , 2010, Future microbiology.

[20]  Thomas Bjarnsholt,et al.  Antibiotic resistance of bacterial biofilms. , 2010, International journal of antimicrobial agents.

[21]  A. Kharazmi,et al.  Polymorphonuclear leucocytes consume oxygen in sputum from chronic Pseudomonas aeruginosa pneumonia in cystic fibrosis , 2009, Thorax.

[22]  N. Høiby,et al.  Pseudomonas aeruginosa biofilms in the respiratory tract of cystic fibrosis patients , 2009, Pediatric pulmonology.

[23]  T. Tolker-Nielsen,et al.  Tolerance to the antimicrobial peptide colistin in Pseudomonas aeruginosa biofilms is linked to metabolically active cells, and depends on the pmr and mexAB‐oprM genes , 2008, Molecular microbiology.

[24]  S. Molin,et al.  In Situ Growth Rates and Biofilm Development of Pseudomonas aeruginosa Populations in Chronic Lung Infections , 2007, Journal of bacteriology.

[25]  J. Collins,et al.  A Common Mechanism of Cellular Death Induced by Bactericidal Antibiotics , 2007, Cell.

[26]  Samuel I. Miller,et al.  Differentiation and Distribution of Colistin- and Sodium Dodecyl Sulfate-Tolerant Cells in Pseudomonas aeruginosa Biofilms , 2006, Journal of bacteriology.

[27]  M. Elkins,et al.  Antibiotic Susceptibilities of Pseudomonas aeruginosa Isolates Derived from Patients with Cystic Fibrosis under Aerobic, Anaerobic, and Biofilm Conditions , 2005, Journal of Clinical Microbiology.

[28]  Philip S. Stewart,et al.  Diffusion in Biofilms , 2003, Journal of bacteriology.

[29]  Y. Urano,et al.  Development of Novel Fluorescence Probes That Can Reliably Detect Reactive Oxygen Species and Distinguish Specific Species* 210 , 2003, The Journal of Biological Chemistry.

[30]  Philip S. Stewart,et al.  Contributions of Antibiotic Penetration, Oxygen Limitation, and Low Metabolic Activity to Tolerance of Pseudomonas aeruginosa Biofilms to Ciprofloxacin and Tobramycin , 2003, Antimicrobial Agents and Chemotherapy.

[31]  George M. Hilliard,et al.  Anaerobic metabolism and quorum sensing by Pseudomonas aeruginosa biofilms in chronically infected cystic fibrosis airways: rethinking antibiotic treatment strategies and drug targets. , 2002, Advanced drug delivery reviews.

[32]  Richard C Boucher,et al.  Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients. , 2002, The Journal of clinical investigation.

[33]  R. Hancock,et al.  Interactions of Bacterial Cationic Peptide Antibiotics with Outer and Cytoplasmic Membranes ofPseudomonas aeruginosa , 2000, Antimicrobial Agents and Chemotherapy.

[34]  E Maier,et al.  Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. , 1999, Biochemistry.

[35]  P. Stewart,et al.  Spatial Physiological Heterogeneity inPseudomonas aeruginosa Biofilm Is Determined by Oxygen Availability , 1998, Applied and Environmental Microbiology.

[36]  R. Baltimore,et al.  Immunohistopathologic localization of Pseudomonas aeruginosa in lungs from patients with cystic fibrosis. Implications for the pathogenesis of progressive lung deterioration. , 1989, The American review of respiratory disease.

[37]  S. Linn,et al.  Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro. , 1988, Science.

[38]  D. Storm,et al.  Inhibition of Escherichia coli growth and respiration by polymyxin B covalently attached to agarose beads. , 1977, Biochemistry.

[39]  G. Mandell,et al.  pO2, pH, and Redox Potential of Experimental Abscesses , 1974, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[40]  L. Lewandowski,et al.  A mutation suppressing streptomycin dependence. I. An effect on ribosome function. , 1967, Journal of molecular biology.

[41]  O. Maaløe,et al.  DNA replication and the division cycle in Escherichia coli , 1967 .

[42]  N. Høiby,et al.  Maintenance treatment of chronic Pseudomonas aeruginosa infection in cystic fibrosis , 2000 .

[43]  D. Hassett,et al.  Pseudomonas aeruginosa biofilm sensitivity to biocides: use of hydrogen peroxide as model antimicrobial agent for examining resistance mechanisms. , 1999, Methods in enzymology.

[44]  J. Noebels.  Ion-Sensitive Intracellular Microelectrodes. How to Make and Use Them , 1979, The Yale Journal of Biology and Medicine.

[45]  D. Sinclair,et al.  Killing by Bactericidal Antibiotics Does Not Depend on Reactive Oxygen Species , 2022 .