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Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates

BACKGROUND: Antimicrobial resistance is a growing threat to public health. Pseudomonas aeruginosa is a relevant pathogen causing human and animal infections, frequently displaying high levels of resistance to commonly used antimicrobials. The increasing difficulty to develop new effective antibiotic...

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Autores principales: Cabassi, Clotilde Silvia, Sala, Andrea, Santospirito, Davide, Alborali, Giovanni Loris, Carretto, Edoardo, Ghibaudo, Giovanni, Taddei, Simone
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5364734/
https://www.ncbi.nlm.nih.gov/pubmed/28335779
http://dx.doi.org/10.1186/s12941-017-0193-1
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author Cabassi, Clotilde Silvia
Sala, Andrea
Santospirito, Davide
Alborali, Giovanni Loris
Carretto, Edoardo
Ghibaudo, Giovanni
Taddei, Simone
author_facet Cabassi, Clotilde Silvia
Sala, Andrea
Santospirito, Davide
Alborali, Giovanni Loris
Carretto, Edoardo
Ghibaudo, Giovanni
Taddei, Simone
author_sort Cabassi, Clotilde Silvia
collection PubMed
description BACKGROUND: Antimicrobial resistance is a growing threat to public health. Pseudomonas aeruginosa is a relevant pathogen causing human and animal infections, frequently displaying high levels of resistance to commonly used antimicrobials. The increasing difficulty to develop new effective antibiotics have discouraged investment in this area and only a few new antibiotics are currently under development. An approach to overcome antibiotic resistance could be based on antimicrobial peptides since they offer advantages over currently used microbicides. METHODS: The antimicrobial activity of the synthetic peptide AMP2041 was evaluated against 49 P. aeruginosa clinical strains with high levels of antimicrobial resistance, isolated from humans (n = 19) and animals (n = 30). In vitro activity was evaluated by a microdilution assay for lethal dose 90% (LD(90)), while the activity over time was performed by time-kill assay with 12.5 µg/ml of AMP2014. Evidences for a direct membrane damage were investigated on P. aeruginosa ATCC 27853 reference strain, on animal isolate PA-VET 38 and on human isolate PA-H 24 by propidium iodide and on P. aeruginosa ATCC 27853 by scanning electron microscopy. RESULTS: AMP2041 showed a dose-dependent activity, with a mean (SEM) LD(90) of 1.69 and 3.3 µg/ml for animal and human strains, respectively. AMP2041 showed microbicidal activity on P. aeruginosa isolates from a patient with cystic fibrosis (CF) and resistance increased from first infection isolate (LD(90) = 0.3 μg/ml) to the mucoid phenotype (LD(90) = 10.4 μg/ml). The time-kill assay showed a time-dependent bactericidal effect of AMP2041 and LD(90) was reached within 20 min for all the strains. The stain-dead assay showed an increasing of membrane permeabilization and SEM analysis revealed holes, dents and bursts throughout bacterial cell wall after 30 min of incubation with AMP2041. CONCLUSIONS: The obtained results assessed for the first time the good antimicrobial activity of AMP2041 on P. aeruginosa strains of human origin, including those deriving from a CF patient. We confirmed the excellent antimicrobial activity of AMP2041 on P. aeruginosa strains derived from dog otitis. We also assessed that AMP2041 antimicrobial activity is linked to changes of the P. aeruginosa cell wall morphology and to the increasing of membrane permeability.
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spelling pubmed-53647342017-03-28 Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates Cabassi, Clotilde Silvia Sala, Andrea Santospirito, Davide Alborali, Giovanni Loris Carretto, Edoardo Ghibaudo, Giovanni Taddei, Simone Ann Clin Microbiol Antimicrob Research BACKGROUND: Antimicrobial resistance is a growing threat to public health. Pseudomonas aeruginosa is a relevant pathogen causing human and animal infections, frequently displaying high levels of resistance to commonly used antimicrobials. The increasing difficulty to develop new effective antibiotics have discouraged investment in this area and only a few new antibiotics are currently under development. An approach to overcome antibiotic resistance could be based on antimicrobial peptides since they offer advantages over currently used microbicides. METHODS: The antimicrobial activity of the synthetic peptide AMP2041 was evaluated against 49 P. aeruginosa clinical strains with high levels of antimicrobial resistance, isolated from humans (n = 19) and animals (n = 30). In vitro activity was evaluated by a microdilution assay for lethal dose 90% (LD(90)), while the activity over time was performed by time-kill assay with 12.5 µg/ml of AMP2014. Evidences for a direct membrane damage were investigated on P. aeruginosa ATCC 27853 reference strain, on animal isolate PA-VET 38 and on human isolate PA-H 24 by propidium iodide and on P. aeruginosa ATCC 27853 by scanning electron microscopy. RESULTS: AMP2041 showed a dose-dependent activity, with a mean (SEM) LD(90) of 1.69 and 3.3 µg/ml for animal and human strains, respectively. AMP2041 showed microbicidal activity on P. aeruginosa isolates from a patient with cystic fibrosis (CF) and resistance increased from first infection isolate (LD(90) = 0.3 μg/ml) to the mucoid phenotype (LD(90) = 10.4 μg/ml). The time-kill assay showed a time-dependent bactericidal effect of AMP2041 and LD(90) was reached within 20 min for all the strains. The stain-dead assay showed an increasing of membrane permeabilization and SEM analysis revealed holes, dents and bursts throughout bacterial cell wall after 30 min of incubation with AMP2041. CONCLUSIONS: The obtained results assessed for the first time the good antimicrobial activity of AMP2041 on P. aeruginosa strains of human origin, including those deriving from a CF patient. We confirmed the excellent antimicrobial activity of AMP2041 on P. aeruginosa strains derived from dog otitis. We also assessed that AMP2041 antimicrobial activity is linked to changes of the P. aeruginosa cell wall morphology and to the increasing of membrane permeability. BioMed Central 2017-03-23 /pmc/articles/PMC5364734/ /pubmed/28335779 http://dx.doi.org/10.1186/s12941-017-0193-1 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Cabassi, Clotilde Silvia
Sala, Andrea
Santospirito, Davide
Alborali, Giovanni Loris
Carretto, Edoardo
Ghibaudo, Giovanni
Taddei, Simone
Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title_full Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title_fullStr Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title_full_unstemmed Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title_short Activity of AMP2041 against human and animal multidrug resistant Pseudomonas aeruginosa clinical isolates
title_sort activity of amp2041 against human and animal multidrug resistant pseudomonas aeruginosa clinical isolates
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5364734/
https://www.ncbi.nlm.nih.gov/pubmed/28335779
http://dx.doi.org/10.1186/s12941-017-0193-1
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