Molecular analysis of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients

Jarych, Dariusz; Augustynowicz–Kopeć, Ewa; Iwańska, Agnieszka; Parniewski, Paweł; Majchrzak, Marta

doi:10.1038/s41598-021-95034-2

Cited by 10 publications

(5 citation statements)

References 30 publications

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“…A study by Jarych et al, irrespective of geographical distance and sample origin (different clinical samples vs. CF patients), supports our findings, suggesting that MLVA is a robust genotyping method applicable for systematic surveys of P. aeruginosa isolates in patients. The study demonstrates the method's effectiveness in discriminating isolates (23).…”

Section: Discussionmentioning

confidence: 82%

Multiple-Locus Variable-Number Tandem Repeat Analysis Genotyping of Biofilm-Producing Pseudomonas aeruginosa Clinical Isolates

Ramazani,

Izadi Amoli,

Taghizadeh Armaki

et al. 2024

Jundishapur J Microbiol

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Background: Pseudomonas aeruginosa (P. aeruginosa) significantly contributes to hospital-acquired infections. Objectives: This study aimed to investigate the genetic diversity of P. aeruginosa strains using multiple-locus variable-number tandem repeat analysis (MLVA) and to explore the relationship between biofilm production and antibiotic resistance. Methods: In this cross-sectional study, 79 P. aeruginosa isolates were collected. Antibiotic sensitivity was tested using the Kirby-Bauer method, and biofilm production capability was assessed through the microtiter plate method. Genetic diversity was evaluated by MLVA, analyzing eight variable-number tandem repeat (VNTR) loci: MS-213, MS-214, MS-207, MS-217, MS-222, MS-209, MS-77, and MS-172. Phylogenetic relationships were delineated using PHYLOViZ 2.0 software. Results: The patient cohort comprised 51.9% males, with the majority of samples (35.4%) obtained from urine. Ceftazidime (CAZ 30µg) showed the highest resistance rate at 77.2%. Notably, 92.4% of isolates were capable of forming biofilms, categorized as 22.7% weak, 28.7% moderate, and 46.5% strong. Phylogenetic analysis demonstrated variability across one or more VNTR loci. Simpson’s index (0.906) and Shannon-Weiner diversity indices (H: 3.466, J: 0.910, Hmax: 3.807, Hmin: 1.242) identified MS77 as the most informative marker for genetic diversity among the isolates. Conclusions: The study highlights an alarming trend in antibiotic resistance, underscoring the necessity of regular monitoring. The findings confirm that MLVA is a straightforward, rapid genotyping method suitable for assessing the genetic diversity of P. aeruginosa.

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Section: Discussionmentioning

confidence: 82%

Multiple-Locus Variable-Number Tandem Repeat Analysis Genotyping of Biofilm-Producing Pseudomonas aeruginosa Clinical Isolates

Ramazani,

Izadi Amoli,

Taghizadeh Armaki

et al. 2024

Jundishapur J Microbiol

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“…The genomes of the underlying P. aeruginosa show little variability in the early stage of a chronic lung infection, but with the duration of the chronic course, single nucleotide mutations and genetic inversions in accessory and housekeeping genes result in diversification. On the other hand, previous studies showed that CF patients can be simultaneously colonized by several different strains that might be residents of the pulmonary tract microbiome prior to an exacerbation [62,63]. All these findings emphasize that antimicrobial resistance testing based solely on a single isolate may lead to ineffective treatment against other coexisting subspecies within the cystic fibrosis (CF) lung.…”

Section: Discussionmentioning

confidence: 99%

The Impact of Pseudomonas aeruginosa Infection in Adult Cystic Fibrosis Patients—A Single Polish Centre Study

Jarzynka,

Makarewicz,

Weiss

et al. 2023

Pathogens

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Background: Pseudomonas aeruginosa (PA) is one of the most predominant pathogens of lung infections, often causing exacerbations in adult patients with cystic fibrosis (CF). Materials and Methods: Microbiological characterization of 74 PA isolates and to evaluate the correlations between the bacterial features and 44 adult Polish CF cohort clinical parameters. Results: The most common variant in the CF transmembrane conductance regulator (CFTR) gene was F508del (76.3%), followed by 3849+10kbC>T (26.3%). A total of 39.4% of the PA isolates showed multiple resistances. In patients with parameters pointing to a decline in lung function, there was a statistically significant moderate correlation with β-lactam resistance and a weak correlation between hospital frequency and colistin resistance. The mucoidity did not correlate with the biofilm formation ability, which showed 41.9% of the isolates. Proteolytic activity, observed in 60.8% of the clinical isolates, was weakly associated with motility detected in 78.4% of the strains. The genetic profiles of the PA were highly heterogeneous, and a weak positive correlation was established between cluster group and biofilm formation. Conclusion: The findings suggest that there is a high variety in P. aeruginosa populations in adult CF patients. There is a need to monitor PA strains in groups of patients with cystic fibrosis, in particular, in terms of the occurrence of antibiotic resistance related to a decline in lung function.

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“…This highlights the well-documented variability in P. aeruginosa strains that occupy the CF lung. This variability underscores the complexity of managing infections in CF patients, as the differences in phage susceptibility and antibiotic resistance can significantly impact treatment efficacy and outcomes (26).…”

Section: Clonality Of the Isolatesmentioning

confidence: 99%

Temperate bacteriophage induced inPseudomonas aeruginosabiofilms can modulate bacteriophage and antibiotic resistance

Martinet,

Samuel,

Weiss

et al. 2024

Preprint

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Given the high levels of resistance in Gram-negative bacteria, phage therapy is garnering increasing attention. In Germany, a clinical study is already underway investigating a phage cocktail for the treatment ofPseudomonas aeruginosain cystic fibrosis (CF) patients. In our study, we examined susceptibility to virulent phages and the PF1-like prophage and antimicrobial profiles and ofP. aeruginosaisolates from a local cystic fibrosis cohort to identify correlations and lysogenic conversion of the prophegs. Consistent with other studies, prophage Pf4 is the most prevalent in this cohort and is activated in the absence of other influences during biofilm formation. These phages can be transferred to other strains that do not contain Pf1-like prophages, thereby influencing the dynamics of bacterial populations in the CF lung. This also rapidly leads to the emergence of a subpopulation resistant to the virulent phages, potentially complicating phage therapy. However, this subset also becomes more susceptible to most antibiotics commonly used in CF, which could be a useful treatment strategy. Interestingly, this bacterial subset lost its susceptibility to colistin, an important inhaled antibiotic in CF, which could lead to treatment failure. Our research highlights both the difficulties and potential strategies to improve treatment options for CF patients.Author summaryWe investigated 51P. aeruginosaisolates obtained from CF patients for the presence of PF1-like prophages and characterized their susceptibility prior and after lysogenig conversion of the prophages to three virulent phages. Our study revealed that the temperate phage Pf4 is the most prominent PF1-like prophage in this cohort, undergoing lysogenic conversion during biofilm formation. The virions identified in the biofilm supernatants are superinfective and transferable to other prophage-freeP. aeruginosaisolates, shaping population dynamics in the CF lung. Prophage reactivation results in the survival of a sub-population with reduced susceptibility to virulent phages, posing a potential challenge for phage therapy. However, this sub-population exhibited restored susceptibility to most CF-relevant antibiotics, presenting an intriguing therapeutic opportunity. Targeted prophage reactivation could sensitize multidrug-resistantP. aeruginosain CF patients, enhancing or even restoring antimicrobial efficacy. Notably, this sub-population also showed a loss of colistin susceptibility, which may lead to treatment failure when colistin is used as an inhaled antipseudomonal antibiotic. Our findings highlight both significant challenges and potential therapeutic opportunities for optimizing the treatment of CF patients. However, these findings are preliminary and require further investigation, particularly regarding the possibility of lysogenic conversion in other prophages (such asSiphoviridae) and how these interplays with resistance to virulent phages. Further studies are ongoing and will enhance our understanding of the role of prophages in the pathophysiology of CF.

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Molecular analysis of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients

Cited by 10 publications

References 30 publications

Multiple-Locus Variable-Number Tandem Repeat Analysis Genotyping of Biofilm-Producing Pseudomonas aeruginosa Clinical Isolates

Multiple-Locus Variable-Number Tandem Repeat Analysis Genotyping of Biofilm-Producing Pseudomonas aeruginosa Clinical Isolates

The Impact of Pseudomonas aeruginosa Infection in Adult Cystic Fibrosis Patients—A Single Polish Centre Study

Temperate bacteriophage induced inPseudomonas aeruginosabiofilms can modulate bacteriophage and antibiotic resistance

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