The effect of alginate lyase on the gentamicin resistance of Pseudomonas aeruginosa in mucoid biofilms

• Published in: Journal of applied microbiology Vol. 121; no. 1; pp. 126 - 135
• Main Authors: Germoni, L.A.P., Bremer, P.J., Lamont, I.L.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-07-2016
• Subjects: alginate lyase; Anti-Bacterial Agents - pharmacology; antibiotic sensitivity; Antibiotics; bacterial alginate; Bacterial infections; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; biofilms; Biofilms - drug effects; cystic fibrosis; Cystic Fibrosis - drug therapy; Cystic Fibrosis - microbiology; Drug resistance; Drug Resistance, Bacterial - drug effects; Gentamicins - pharmacology; Humans; Microbiology; Polysaccharide-Lyases - genetics; Polysaccharide-Lyases - metabolism; Pseudomonas aeruginosa; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - physiology; Pseudomonas Infections - drug therapy; Pseudomonas Infections - microbiology; alginate lyase; Pseudomonas aeruginosa; bacterial alginate; biofilms; cystic fibrosis; antibiotic sensitivity
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.13153

Aims
Pseudomonas aeruginosa can secrete large amounts of alginate during chronic infections and this has been associated with high resistance to antibiotics. The major aim of this study was to investigate whether degradation of extracellular alginate by alginate lyase would increase the sensitivity of Ps. aeruginosa to gentamicin, an aminoglycoside antibiotic.
Methods and Results
Degradation of alginate from Ps. aeruginosa was monitored using a spectrometric assay. Alginate lyase depolymerized alginate, but calcium and zinc cations at concentrations found in the cystic fibrosis lung reduced enzyme activity. Biofilms formed on agar were partially degraded by alginate lyase, but staining with crystal violet showed that the biomass of biofilms grown in liquid was not significantly affected by the enzyme. Viability testing showed that the sensitivity to gentamicin of biofilm bacteria and of bacteria released from biofilms was unaffected by alginate lyase.
Conclusions
Our results show that at least under the conditions used here alginate lyase does not affect gentamicin resistance of Ps. aeruginosa.
Significance and Impact of the Study
Our study indicates that alginate does not contribute to resistance to gentamicin and so does not provide support for the concept of treating patients with alginate lyase in order to increase the antibiotic sensitivity of Ps. aeruginosa.