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. 2023 Dec 19;13(1):1.
doi: 10.3390/antibiotics13010001.

Virulence of Pseudomonas aeruginosa in Cystic Fibrosis: Relationships between Normoxia and Anoxia Lifestyle

Rosanna Papa  1 , Esther Imperlini  2 , Marika Trecca  1 , Irene Paris  1 , Gianluca Vrenna  3 , Marco Artini  1 , Laura Selan  1
Affiliations

Virulence of Pseudomonas aeruginosa in Cystic Fibrosis: Relationships between Normoxia and Anoxia Lifestyle

Rosanna Papa et al. Antibiotics (Basel). .

Abstract

The airways of cystic fibrosis (CF) patients are colonized by many pathogens and the most common is Pseudomonas aeruginosa, an environmental pathogen that is able to infect immunocompromised patients thanks to its ability to develop resistance to conventional antibiotics. Over 12% of all patients colonized by P. aeruginosa harbour multi-drug resistant species. During airway infection in CF, P. aeruginosa adopts various mechanisms to survive in a hostile ecological niche characterized by low oxygen concentration, nutrient limitation and high osmotic pressure. To this end, P. aeruginosa uses a variety of virulence factors including pigment production, biofilm formation, motility and the secretion of toxins and proteases. This study represents the first report that systematically analyzes the differences in virulence features, in normoxia and anoxia, of clinical P. aeruginosa isolated from CF patients, characterized by multi- or pan-drug antibiotic resistance compared to antibiotic sensitive strains. The virulence features, such as biofilm formation, protease secretion and motility, are highly diversified in anaerobiosis, which reflects the condition of chronic CF infection. These findings may contribute to the understanding of the real-world lifestyle of pathogens isolated during disease progression in each particular patient and to assist in the design of therapeutic protocols for personalized medicine.

Keywords: antimicrobial resistance; biofilm; motility; oxygen concentration; proteases; pyocyanin; pyoverdine.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Pyocyanin production of WT, MDR and PDR strains at 24 h and 48 h in normoxia. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 2
Figure 2
Pyoverdine production of WT, MDR and PDR strains at 24 h, 48 h and 72 h. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 3
Figure 3
Biofilm formation of WT, MDR and PDR strains in normoxia and anoxia. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 4
Figure 4
Protease production of WT, MDR and PDR strains at 24 h and 48 h in normoxia and anoxia. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 5
Figure 5
Gelatin-zymography to assay proteases secreted by P. aeruginosa clinical strains at 24 h and 48 h in normoxia and anoxia. Protein samples from unconcentrated culture supernatants were separated by using 10% SDS-PAGE gel with 0.2% gelatin. All gels were incubated in a development buffer and destained with 0.5% Coomassie blue R-250, thus obtaining clear bands corresponding to active proteases on a blue background. MW, molecular weight in kDa of protein markers. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 6
Figure 6
Motility assay: swimming assay of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 6
Figure 6
Motility assay: swimming assay of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 6
Figure 6
Motility assay: swimming assay of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 7
Figure 7
Swarming motility of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 7
Figure 7
Swarming motility of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
Figure 7
Figure 7
Swarming motility of P. aeruginosa bacterial strains in normoxia (left panel) and anoxia (right panel) measured at 24 h, 48 h and 72 h of bacterial growth. WT: wild type, sensitive strains; MDR: multi-drug resistant strains; PDR: pan-drug resistant strains.
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