Effects of a novel anti-biofilm peptide CRAMP combined with antibiotics on the formation of Pseudomonas aeruginosa biofilms

Zhang, Yang; He, Xinkui; Peng, Cheng; Li, Xiaofen; Wang, Shiyuan; Xiong, Jing; Li, Hui; Wang, Zhiying; Yi, Huashan; Liu, Juan; Chen, Hongwei

doi:10.1016/j.micpath.2020.104660

Cited by 12 publications

(9 citation statements)

References 31 publications

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“…In recent years, the use of dispersants combined with antibiotics has been reported, and this combination is highly advantageous [ 59 – 61 ]. In our previous study, it was shown that the combination of subinhibitory concentrations of CRAMP and colistin has a significant synergistic effect on the formation of PAO1 biofilms [ 19 ], and we also recently found that CRAMP combined with vancomycin, roxithromycin, and azithromycin showed faster and stronger antibiofilm bacterial activity than that of a single drug through the time-kill curve test, especially combination with vancomycin, which caused the biofilm cells to die within 3 h (a manuscript is in the preparation stage). These studies demonstrate that it is promising to develop CRAMP as a potential biofilm dispersant.…”

Section: Discussionmentioning

confidence: 99%

“…The morphological features of biofilms were observed by CLSM as described previously with some modifications [ 19 ]. In this experiment, 250 µL of the undiluted test bacterial solution ( OD 600 = 0.1) was added to an 8-well chambered cover glass (1.5 Borosilicate glass, Lab-Tek II chambered coverglass, Rochester, NY, USA), and the medium was replaced with fresh medium every 24 h. After incubation for 3 days at 37 ℃, the biofilm was treated with CRAMP at 37 ℃ for 1 h. Then, the biofilm was washed with 0.9% (wt/vol) NaCl and stained for 20 min in the dark at room temperature using a Filmtracer™ LIVE/DEAD™ Biofilm Viability kit (Cat.…”

Section: Methodsmentioning

confidence: 99%

“…However, few studies have concentrated on the mechanism by which AMPs disperse biofilms. We have recently reported that the mouse homologue cathelicidin-related AMP (CRAMP) could inhibit the formation of P. aeruginosa PAO1 biofilms at subminimum inhibitory concentrations (MICs) [ 19 ] and reduce the biomass of preformed P. aeruginosa 27 853 biofilms [ 20 ]. More importantly, in our studies, CRAMP was the only AMP that could eradicate the P. aeruginosa biofilm, while others could only kill part of the bacterial biofilm [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

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Pseudomonas aeruginosa biofilm dispersion by the mouse antimicrobial peptide CRAMP

Zhang

Peng²,

Wang³

et al. 2022

Vet Res

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Pseudomonas aeruginosa (P. aeruginosa) is a known bacterium that produces biofilms and causes severe infection. Furthermore, P. aeruginosa biofilms are extremely difficult to eradicate, leading to the development of chronic and antibiotic-resistant infections. Our previous study showed that a cathelicidin-related antimicrobial peptide (CRAMP) inhibits the formation of P. aeruginosa biofilms and markedly reduces the biomass of preformed biofilms, while the mechanism of eradicating bacterial biofilms remains elusive. Therefore, in this study, the potential mechanism by which CRAMP eradicates P. aeruginosa biofilms was investigated through an integrative analysis of transcriptomic, proteomic, and metabolomic data. The omics data revealed CRAMP functioned against P. aeruginosa biofilms by different pathways, including the Pseudomonas quinolone signal (PQS) system, cyclic dimeric guanosine monophosphate (c-di-GMP) signalling pathway, and synthesis pathways of exopolysaccharides and rhamnolipid. Moreover, a total of 2914 differential transcripts, 785 differential proteins, and 280 differential metabolites were identified. A series of phenotypic validation tests demonstrated that CRAMP reduced the c-di-GMP level with a decrease in exopolysaccharides, especially alginate, in P. aeruginosa PAO1 biofilm cells, improved bacterial flagellar motility, and increased the rhamnolipid content, contributing to the dispersion of biofilms. Our study provides new insight into the development of CRAMP as a potentially effective antibiofilm dispersant.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Pseudomonas aeruginosa biofilm dispersion by the mouse antimicrobial peptide CRAMP

Zhang

Peng²,

Wang³

et al. 2022

Vet Res

Self Cite

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“…In the quantitative assay, proteolytic activity was detected using azocasein as the protease substrate [ 24 ]. After digestion by proteases, the produced protein fragments were quantitatively analyzed by measuring their absorbance, after the removal of the undigested substrate by trichloroacetic acid precipitation.…”

Section: Resultsmentioning

confidence: 99%

Anti-Virulence Potential of a Chionodracine-Derived Peptide against Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates from Cystic Fibrosis Patients

Artini

Imperlini

Buonocore

et al. 2022

IJMS

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Pseudomonas aeruginosa is an opportunistic pathogen causing several chronic infections resistant to currently available antibiotics. Its pathogenicity is related to the production of different virulence factors such as biofilm and protease secretion. Pseudomonas communities can persist in biofilms that protect bacterial cells from antibiotics. Hence, there is a need for innovative approaches that are able to counteract these virulence factors, which play a pivotal role, especially in chronic infections. In this context, antimicrobial peptides are emerging drugs showing a broad spectrum of antibacterial activity. Here, we tested the anti-virulence activity of a chionodracine-derived peptide (KHS-Cnd) on five P. aeruginosa clinical isolates from cystic fibrosis patients. We demonstrated that KHS-Cnd impaired biofilm development and caused biofilm disaggregation without affecting bacterial viability in nearly all of the tested strains. Ultrastructural morphological analysis showed that the effect of KHS-Cnd on biofilm could be related to a different compactness of the matrix. KHS-Cnd was also able to reduce adhesion to pulmonary cell lines and to impair the invasion of host cells by P. aeruginosa. A cytotoxic effect of KHS-Cnd was observed only at the highest tested concentration. This study highlights the potential of KHS-Cnd as an anti-biofilm and anti-virulence molecule against P. aeruginosa clinical strains.

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“…The morphological features of biofilms were observed by CLSM as described previously with some modifications [ 46 ]. In this experiment, 100 μL of the undiluted test bacterial solution ( OD600 = 0.1) was added to an 8-well chambered cover glass (1.5 Borosilicate glass, Lab-Tek II chambered coverglass).…”

Section: Methodsmentioning

confidence: 99%

Effect of baicalin on eradicating biofilms of bovine milk derived Acinetobacter lwoffii

Ma,

Mei,

Liu

et al. 2024

BMC Vet Res

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Background Acinetobacter lwoffii (A.lwoffii) is a serious zoonotic pathogen that has been identified as a cause of infections such as meningitis, bacteremia and pneumonia. In recent years, the infection rate and detection rate of A.lwoffii is increasing, especially in the breeding industry. Due to the presence of biofilms, it is difficult to eradicate and has become a potential super drug-resistant bacteria. Therefore, eradication of preformed biofilm is an alternative therapeutic action to control A.lwoffii infection. The present study aimed to clarify that baicalin could eradicate A.lwoffii biofilm in dairy cows, and to explore the mechanism of baicalin eradicating A.lwoffii. Results The results showed that compared to the control group, the 4 MIC of baicalin significantly eradicated the preformed biofilm, and the effect was stable at this concentration, the number of viable bacteria in the biofilm was decreased by 0.67 Log10CFU/mL. The total fluorescence intensity of biofilm bacteria decreased significantly, with a reduction rate of 67.0%. There were 833 differentially expressed genes (367 up-regulated and 466 down-regulated), whose functions mainly focused on oxidative phosphorylation, biofilm regulation system and trehalose synthesis. Molecular docking analysis predicted 11 groups of target proteins that were well combined with baicalin, and the content of trehalose decreased significantly after the biofilm of A.lwoffii was treated with baicalin. Conclusions The present study evaluated the antibiofilm potential of baicalin against A.lwoffii. Baicalin revealed strong antibiofilm potential against A.lwoffii. Baicalin induced biofilm eradication may be related to oxidative phosphorylation and TCSs. Moreover, the decrease of trehalose content may be related to biofilm eradication.

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Effects of a novel anti-biofilm peptide CRAMP combined with antibiotics on the formation of Pseudomonas aeruginosa biofilms

Cited by 12 publications

References 31 publications

Pseudomonas aeruginosa biofilm dispersion by the mouse antimicrobial peptide CRAMP

Pseudomonas aeruginosa biofilm dispersion by the mouse antimicrobial peptide CRAMP

Anti-Virulence Potential of a Chionodracine-Derived Peptide against Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates from Cystic Fibrosis Patients

Effect of baicalin on eradicating biofilms of bovine milk derived Acinetobacter lwoffii

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