Real-time monitoring of<i>Pseudomonas aeruginosa</i>biofilm growth dynamics and persister cells’ eradication

Žiemytė, Miglė; Carda‐Diéguez, Miguel; Rodríguez-Díaz, Juan Carlos; Ventero, María Paz; Mira, Álex; Ferrer, Milagros

doi:10.1080/22221751.2021.1994355

Cited by 26 publications

(22 citation statements)

References 52 publications

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“…All the tested isolates were strong biofilm formers; however, the persister isolates had stronger biofilms (as their OD was twice that of the PAO1 strain). In contrast, a previous study reported that the biofilm of PAO1 and their persister strain had the same strength [ 26 ].…”

Section: Discussionmentioning

confidence: 79%

“…Previous reports have suggested that persister cells exhibit different phenotypes compared to the wild-type strains, including smaller colony sizes or changes in pigmentation [ 10 , 14 , 26 ]. Given that persister cells can enter a non-growing state, tolerate high concentrations of bactericidal antibiotics (for example by activating efflux pumps), and regrow once the treatment is ceased, these cells have already been linked to the relapse and recalcitrance of chronic infections [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

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Genome Analysis of Pseudomonas aeruginosa Strains from Chronically Infected Patients with High Levels of Persister Formation

et al. 2023

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The appearance of persister cells with low metabolic rates are key factors leading to antibiotic treatment failure. Such persisters are multidrug tolerant and play a key role in the recalcitrance of biofilm-based chronic infections. Here, we present the genomic analyses of three distinct Pseudomonas aeruginosa Egyptian persister-isolates recovered from chronic human infections. To calculate the persister frequencies, viable counts were determined before and after treatment with levofloxacin. The susceptibilities of isolates to different antibiotics were determined using the agar-dilution method. To determine their recalcitrance, the levofloxacin persisters were further challenged with lethal concentrations of meropenem, tobramycin, or colistin. Furthermore, the biofilm formation of the persister strains was estimated phenotypically, and they were reported to be strong biofilm-forming strains. The genotypic characterization of the persisters was performed using whole genome sequencing (WGS) followed by phylogenetic analysis and resistome profiling. Interestingly, out of the thirty-eight clinical isolates, three isolates (8%) demonstrated a persister phenotype. The three levofloxacin-persister isolates were tested for their susceptibility to selected antibiotics; all of the tested isolates were multidrug resistant (MDR). Additionally, the P. aeruginosa persisters were capable of surviving over 24 h and were not eradicated after exposure to 100X-MIC of levofloxacin. WGS for the three persisters revealed a smaller genome size compared to PAO1-genome. Resistome profiling indicated the presence of a broad collection of antibiotic-resistance genes, including genes encoding for antibiotic-modifying enzymes and efflux pump. Phylogenetic analysis indicated that the persister isolates belong to a distinct clade rather than the deposited P. aeruginosa strains in the GenBank. Conclusively, the persister isolates in our study are MDR and form a highly strong biofilm. WGS revealed a smaller genome that belongs to a distinct clade.

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Section: Discussionmentioning

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Genome Analysis of Pseudomonas aeruginosa Strains from Chronically Infected Patients with High Levels of Persister Formation

et al. 2023

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“…Understanding these pleiotropic interactions would be very useful to optimize the treatments of infections or cancers [66]. Indeed, this might allow to pre-evolve an infecting bacterial cell population or a cancer cell population to have a low persistence to the future treatment, or to be more sensitive to molecules such as mannitol that starts being used against persistent cells [66,67].…”

Section: Discussionmentioning

confidence: 99%

Evolution of bacterial persistence to antibiotics during a 50,000-generation experiment in an antibiotic-free environment

Mathe‐Hubert

Amia

Martin

et al. 2022

Preprint

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Failure of antibiotic therapies causes > 700,000 deaths yearly and involves both bacterial resistance and persistence. Persistence results in the relapse of infections by producing a tiny fraction of pathogen survivors that stay dormant during antibiotic exposure. From an evolutionary perspective, persistence is either a 'bethedging strategy' that helps to cope with stochastically changing environments or an unavoidable minimal rate of 'cellular errors' that lock the cells in a low activity state. Here, we analyzed the evolution of persistence over 50,000 bacterial generations in a stable environment by improving a published method that estimates the number of persister cells based on the growth of the reviving population. Our results challenged our understanding of the factors underlying persistence evolution. In one case, we observed a substantial decrease in persistence proportion, suggesting that the naturally observed persistence level is not an unavoidable minimal rate of 'cellular errors'. However, although there was no obvious environmental stochasticity, in most cases the persistence level was maintained during 50,000 bacterial generations, and even increased in few cases.

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“…To assess the inhibition of biofilm formation, a crystal violet staining assay was conducted as per a previously published protocol, albeit with a slight modification . After overnight incubation, individual colony isolates were adjusted to 0.5 McFarland with NS.…”

Section: Materials and Methodsmentioning

confidence: 99%

Restoring Colistin Sensitivity and Combating Biofilm Formation: Synergistic Effects of Colistin and Usnic Acid against Colistin-Resistant Enterobacteriaceae

Zhang,

Han,

Huang

et al. 2023

ACS Infect. Dis.

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Real-time monitoring ofPseudomonas aeruginosabiofilm growth dynamics and persister cells’ eradication

Cited by 26 publications

References 52 publications

Genome Analysis of Pseudomonas aeruginosa Strains from Chronically Infected Patients with High Levels of Persister Formation

Genome Analysis of Pseudomonas aeruginosa Strains from Chronically Infected Patients with High Levels of Persister Formation

Evolution of bacterial persistence to antibiotics during a 50,000-generation experiment in an antibiotic-free environment

Restoring Colistin Sensitivity and Combating Biofilm Formation: Synergistic Effects of Colistin and Usnic Acid against Colistin-Resistant Enterobacteriaceae

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