Increased Susceptibility to Azithromycin of Pseudomonas Aeruginosa Biofilms Using RPMI 1640 Testing Media

Overview

Journal APMIS

Specialties Allergy & Immunology
Microbiology
Pathology

Date 2024 Apr 16

PMID 38622982

Authors

Adrian Jimenez San San Mauro

Niels Hoiby

Oana Ciofu

Affiliations

Soon will be listed here.

Abstract

Azithromycin (AZM) is efficient for treatment of chronic Pseudomonas aeruginosa biofilm lung infections, despite of resistance in conventional susceptibility testing. It has been shown that planktonic P. aeruginosa are more susceptible to AZM when tested in RPMI 1640 medium. The aim of the study was to test the susceptibility to AZM of P. aeruginosa biofilms in LB vs RPMI 1640 media. We investigated the effect of AZM on planktonic and biofilms of (WT) P. aeruginosa (PAO1), the hypermutable (ΔmutS) and the antibiotic-resistant phenotype(ΔnfxB) mutants. The effect of AZM on young and mature biofilms was investigated in the modified Calgary Biofilm Device by estimation of the minimal biofilm inhibitory concentration (MBIC). The AZM MBIC in LB/RPMI1640 on young biofilms treated for 24 h was 16/4 μg/mL for PAO1, 32/8 μg/mL for ΔmutS, and 256/16 μg/mL for ΔnfxB, while in mature biofilms was 256/2 μg/mL for PAO1 and ΔmutS and 16/1 μg/mL for ΔnfxB. The effect of AZM was improved when the treatment was prolonged to 72 h, supporting the intracellular accumulation of AZM. An increased susceptibility of P. aeruginosa biofilms to AZM was observed in RPMI 1640 than in LB medium. Our results might improve susceptibility testing and dosing of AZM for treatment of biofilm infections.

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References

Wilms E, Touw D, Heijerman H . Pharmacokinetics of azithromycin in plasma, blood, polymorphonuclear neutrophils and sputum during long-term therapy in patients with cystic fibrosis. Ther Drug Monit. 2006; 28(2):219-25. DOI: 10.1097/01.ftd.0000195617.69721.a5. View

Lee S, Gallagher L, Manoil C . Reconstructing a Wild-Type Pseudomonas aeruginosa Reference Strain PAO1. J Bacteriol. 2021; 203(14):e0017921. PMC: 8223932. DOI: 10.1128/JB.00179-21. View

Lai P, Schibler M, Walters J . Azithromycin enhances phagocytic killing of Aggregatibacter actinomycetemcomitans Y4 by human neutrophils. J Periodontol. 2014; 86(1):155-61. PMC: 4543394. DOI: 10.1902/jop.2014.140183. View

Parnham M, Erakovic Haber V, Giamarellos-Bourboulis E, Perletti G, Verleden G, Vos R . Azithromycin: mechanisms of action and their relevance for clinical applications. Pharmacol Ther. 2014; 143(2):225-45. DOI: 10.1016/j.pharmthera.2014.03.003. View

Swatton J, Davenport P, Maunders E, Griffin J, Lilley K, Welch M . Impact of Azithromycin on the Quorum Sensing-Controlled Proteome of Pseudomonas aeruginosa. PLoS One. 2016; 11(1):e0147698. PMC: 4726577. DOI: 10.1371/journal.pone.0147698. View

Altenburg J, de Graaff C, Stienstra Y, Sloos J, van Haren E, Koppers R . Effect of azithromycin maintenance treatment on infectious exacerbations among patients with non-cystic fibrosis bronchiectasis: the BAT randomized controlled trial. JAMA. 2013; 309(12):1251-9. DOI: 10.1001/jama.2013.1937. View

Ciofu O, Mandsberg L, Bjarnsholt T, Wassermann T, Hoiby N . Genetic adaptation of Pseudomonas aeruginosa during chronic lung infection of patients with cystic fibrosis: strong and weak mutators with heterogeneous genetic backgrounds emerge in mucA and/or lasR mutants. Microbiology (Reading). 2009; 156(Pt 4):1108-1119. DOI: 10.1099/mic.0.033993-0. View

Hengzhuang W, Hoiby N, Ciofu O . Pharmacokinetics and pharmacodynamics of antibiotics in biofilm infections of Pseudomonas aeruginosa in vitro and in vivo. Methods Mol Biol. 2014; 1147:239-54. DOI: 10.1007/978-1-4939-0467-9_17. View

Oliver A, Canton R, Campo P, Baquero F, Blazquez J . High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science. 2000; 288(5469):1251-4. DOI: 10.1126/science.288.5469.1251. View

10.

Ersoy S, Heithoff D, Barnes 5th L, Tripp G, House J, Marth J . Correcting a Fundamental Flaw in the Paradigm for Antimicrobial Susceptibility Testing. EBioMedicine. 2017; 20:173-181. PMC: 5478264. DOI: 10.1016/j.ebiom.2017.05.026. View

11.

Hansen C, Pressler T, Koch C, Hoiby N . Long-term azitromycin treatment of cystic fibrosis patients with chronic Pseudomonas aeruginosa infection; an observational cohort study. J Cyst Fibros. 2005; 4(1):35-40. DOI: 10.1016/j.jcf.2004.09.001. View

12.

Jaffe A, Bush A . Anti-inflammatory effects of macrolides in lung disease. Pediatr Pulmonol. 2001; 31(6):464-73. DOI: 10.1002/ppul.1076. View

13.

Imamura Y, Higashiyama Y, Tomono K, Izumikawa K, Yanagihara K, Ohno H . Azithromycin exhibits bactericidal effects on Pseudomonas aeruginosa through interaction with the outer membrane. Antimicrob Agents Chemother. 2005; 49(4):1377-80. PMC: 1068619. DOI: 10.1128/AAC.49.4.1377-1380.2005. View

14.

Marvig R, Sondergaard M, Damkiaer S, Hoiby N, Krogh Johansen H, Molin S . Mutations in 23S rRNA confer resistance against azithromycin in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 2012; 56(8):4519-21. PMC: 3421623. DOI: 10.1128/AAC.00630-12. View

15.

Jorgensen K, Wassermann T, Jensen P, Hengzuang W, Molin S, Hoiby N . Sublethal ciprofloxacin treatment leads to rapid development of high-level ciprofloxacin resistance during long-term experimental evolution of Pseudomonas aeruginosa. Antimicrob Agents Chemother. 2013; 57(9):4215-21. PMC: 3754285. DOI: 10.1128/AAC.00493-13. View

16.

Goltermann L, Andersen K, Krogh Johansen H, Molin S, La Rosa R . Macrolide therapy in Pseudomonas aeruginosa infections causes uL4 ribosomal protein mutations leading to high-level resistance. Clin Microbiol Infect. 2022; 28(12):1594-1601. DOI: 10.1016/j.cmi.2022.08.003. View

17.

Saiman L, Marshall B, Mayer-Hamblett N, Burns J, Quittner A, Cibene D . Azithromycin in patients with cystic fibrosis chronically infected with Pseudomonas aeruginosa: a randomized controlled trial. JAMA. 2003; 290(13):1749-56. DOI: 10.1001/jama.290.13.1749. View

18.

Kobbernagel H, Buchvald F, Haarman E, Casaulta C, Collins S, Hogg C . Efficacy and safety of azithromycin maintenance therapy in primary ciliary dyskinesia (BESTCILIA): a multicentre, double-blind, randomised, placebo-controlled phase 3 trial. Lancet Respir Med. 2020; 8(5):493-505. DOI: 10.1016/S2213-2600(20)30058-8. View

19.

Oliver A, Mena A . Bacterial hypermutation in cystic fibrosis, not only for antibiotic resistance. Clin Microbiol Infect. 2010; 16(7):798-808. DOI: 10.1111/j.1469-0691.2010.03250.x. View

20.

Buyck J, Plesiat P, Traore H, Vanderbist F, Tulkens P, Van Bambeke F . Increased susceptibility of Pseudomonas aeruginosa to macrolides and ketolides in eukaryotic cell culture media and biological fluids due to decreased expression of oprM and increased outer-membrane permeability. Clin Infect Dis. 2012; 55(4):534-42. DOI: 10.1093/cid/cis473. View