Membrane phospholipid composition of Pseudomonas aeruginosa grown in a cystic fibrosis mucus-mimicking medium

Deschamps, Estelle; Schaumann, Annick; Schmitz-Afonso, Isabelle; Afonso, Carlos; Dé, Emmanuelle; Loutelier‐Bourhis, Corinne; Alexandre, Stéphane

doi:10.1016/j.bbamem.2020.183482

Cited by 17 publications

(21 citation statements)

References 39 publications

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“…Pseudomonas bacteria were shown to have a different pattern of gene expression and phenotypic characteristics in lung sputum or in media mimicking the composition of CF sputum compared to when grown in standard laboratory media . This includes changes in membrane composition, which can affect antibiotic susceptibility . Since the media can have an effect on the bacterial physiology and on the antimicrobial activity, we investigated the antimicrobial activity under standard conditions and with growth media that mimic the environment of two types of infections: (i) a chronic wound infection using SWF and (ii) a respiratory infection of a CF lung using SCFM …”

Section: Resultsmentioning

confidence: 99%

“…34 This includes changes in membrane composition, which can affect antibiotic susceptibility. 35 Since the media can have an effect on the bacterial physiology and on the antimicrobial activity, we investigated the antimicrobial activity under standard conditions and with growth media that mimic the environment of two types of infections: (i) a chronic wound infection using SWF 36 and (ii) a respiratory infection of a CF lung using SCFM. 34 We used two isolates of each species.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Evaluation of the Antimicrobial Activity in Host-Mimicking Media and In Vivo Toxicity of Antimicrobial Polymers as Functional Mimics of AMPs

Maset

Hapeshi

Hall

et al. 2022

ACS Appl. Mater. Interfaces

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Activity tests for synthetic antimicrobial compounds are often limited to the minimal inhibitory concentration assay using standard media and bacterial strains. In this study, a family of acrylamide copolymers that act as synthetic mimics of antimicrobial peptides were synthesized and shown to have a disruptive effect on bacterial membranes and structural integrity through microscopy techniques and membrane polarization experiments. The polymers were tested for their antimicrobial properties using media that mimic clinically relevant conditions. Additionally, their activity was compared in two different strains of the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Pseudomonas aeruginosa. We showed that the medium composition can have an important influence on the polymer activity as there was a considerable reduction in minimal inhibitory concentrations against S. aureus grown in synthetic wound fluid (SWF), and against P. aeruginosa grown in synthetic cystic fibrosis sputum media (SCFM), compared to the concentrations in standard testing media. In contrast, we observed a complete loss of activity against P. aeruginosa in the serum-containing SWF. Finally, we made use of an emerging invertebrate in vivo model, using Galleria mellonella larvae, to assess toxicity of the polymeric antimicrobials, showing a good correlation with cell line toxicity measurements and demonstrating its potential in the evaluation of novel antimicrobial materials.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Evaluation of the Antimicrobial Activity in Host-Mimicking Media and In Vivo Toxicity of Antimicrobial Polymers as Functional Mimics of AMPs

Maset

Hapeshi

Hall

et al. 2022

ACS Appl. Mater. Interfaces

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show abstract

“…Pseudomonas aeruginosa PAO1 was cultivated in the defined rich medium Synthetic Cystic Fibrosis Medium (SCFM), designed to mimic the conditions of the airways in patients with cystic fibrosis and containing amino acids, glucose and lactate as carbon sources (Turner et al ., 2015). We excluded DNA and mucins to reduce viscosity and allow HPLC analysis of the samples (Deschamps et al ., 2021). Two different batches of SCFM were used for the cultivation experiments in deep‐well plates and Erlenmeyer flasks, respectively, and some variation in the concentration of specific metabolites was determined by HPLC analysis.…”

Section: Methodsmentioning

confidence: 99%

High‐throughput dilution‐based growth method enables time‐resolved exo‐metabolomics of Pseudomonas putida and Pseudomonas aeruginosa

Pedersen

Gurdo

Johansen

et al. 2021

Microbial Biotechnology

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Understanding metabolism is fundamental to access and harness bacterial physiology. In most bacteria, nutrient utilization is hierarchically optimized according to their energetic potential and their availability in the environment to maximise growth rates. Lowthroughput methods have been largely used to characterize bacterial metabolic profiles. However, indepth analysis of large collections of strains across several conditions is challenging since highthroughput approaches are still limitedespecially for non-traditional hosts. Here, we developed a highthroughput dilution-resolved cultivation method for metabolic footprinting of Pseudomonas putida and Pseudomonas aeruginosa. This method was benchmarked against a conventional low-throughput timeresolved cultivation approach using either a synthetic culture medium (where a single carbon source is present) for P. putida or a complex nutrient mixture for P. aeruginosa. Dynamic metabolic footprinting, either by sugar quantification or by targeted exo-metabolomic analyses, revealed overlaps between the bacterial metabolic profiles irrespective of the cultivation strategy, suggesting a certain level of robustness and flexibility of the high-throughput dilution-resolved method. Cultivation of P. putida in microtiter plates imposed a metabolic constraint, dependent on oxygen availability, which altered the pattern of secreted metabolites at the level of sugar oxidation. Deep-well plates, however, constituted an optimal cultivation set-up yielding consistent and comparable metabolic profiles across conditions and strains. Altogether, the results illustrate the usefulness of this technological advance for highthroughput analyses of bacterial metabolism for both biotechnological applications and automation purposes.

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“…For example, P. aeruginosa grown in synthetic cystic brosis media (SCFM) showed lower levels of PC compared to P. aeruginosa grown in Muller-Hilton broth (MHB). 64 To more accurately simulate the natural biological environment, the phospholipid composition of the bacterial were also studied in SCFM with added DOPC (SCFM-PC). 65 As with SCFM only, there was a general increase in comparative quantities of those lipids present within P. aeruginosa grown in SCFM-PC, compared to that grown in MHB.…”

Section: Phospholipid Composition Of Bacterial Membranesmentioning

confidence: 99%

The phospholipid membrane compositions of bacterial cells, cancer cell lines and biological samples from cancer patients

et al. 2021

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Membrane phospholipid composition of Pseudomonas aeruginosa grown in a cystic fibrosis mucus-mimicking medium

Cited by 17 publications

References 39 publications

Evaluation of the Antimicrobial Activity in Host-Mimicking Media and In Vivo Toxicity of Antimicrobial Polymers as Functional Mimics of AMPs

Evaluation of the Antimicrobial Activity in Host-Mimicking Media and In Vivo Toxicity of Antimicrobial Polymers as Functional Mimics of AMPs

High‐throughput dilution‐based growth method enables time‐resolved exo‐metabolomics of Pseudomonas putida and Pseudomonas aeruginosa

The phospholipid membrane compositions of bacterial cells, cancer cell lines and biological samples from cancer patients

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