A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo
The opportunistic human pathogen <i>Pseudomonas aeruginosa</i> is responsible for a variety of acute infections and is a major cause of mortality in chronically infected cystic fibrosis patients. Due to increased resistance to antibiotics, new therapeutic strategies against <i>P. a...
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MDPI AG
2021-03-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/4/401 |
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| author | Pauline Nogaret Fatima El Garah Anne-Béatrice Blanc-Potard |
| author_facet | Pauline Nogaret Fatima El Garah Anne-Béatrice Blanc-Potard |
| author_sort | Pauline Nogaret |
| collection | DOAJ |
| description | The opportunistic human pathogen <i>Pseudomonas aeruginosa</i> is responsible for a variety of acute infections and is a major cause of mortality in chronically infected cystic fibrosis patients. Due to increased resistance to antibiotics, new therapeutic strategies against <i>P. aeruginosa</i> are urgently needed. In this context, we aimed to develop a simple vertebrate animal model to rapidly assess in vivo drug efficacy against <i>P. aeruginosa</i>. Zebrafish are increasingly considered for modeling human infections caused by bacterial pathogens, which are commonly microinjected in embryos. In the present study, we established a novel protocol for zebrafish infection by <i>P. aeruginosa</i> based on bath immersion in 96-well plates of tail-injured embryos. The immersion method, followed by a 48-hour survey of embryo viability, was first validated to assess the virulence of <i>P. aeruginosa</i> wild-type PAO1 and a known attenuated mutant. We then validated its relevance for antipseudomonal drug testing by first using a clinically used antibiotic, ciprofloxacin. Secondly, we used a novel quorum sensing (QS) inhibitory molecule, <i>N</i>-(2-pyrimidyl)butanamide (C11), the activity of which had been validated in vitro but not previously tested in any animal model. A significant protective effect of C11 was observed on infected embryos, supporting the ability of C11 to attenuate in vivo <i>P. aeruginosa</i> pathogenicity. In conclusion, we present here a new and reliable method to compare the virulence of <i>P. aeruginosa</i> strains in vivo and to rapidly assess the efficacy of clinically relevant drugs against <i>P. aeruginosa</i>, including new antivirulence compounds. |
| first_indexed | 2024-03-10T12:48:47Z |
| format | Article |
| id | doaj.art-dba9fb5efdc14376abed330e29d280c4 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T12:48:47Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-dba9fb5efdc14376abed330e29d280c42023-11-21T13:15:02ZengMDPI AGPathogens2076-08172021-03-0110440110.3390/pathogens10040401A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In VivoPauline Nogaret0Fatima El Garah1Anne-Béatrice Blanc-Potard2Laboratory of Pathogen-Host Interactions (LPHI), Université Montpellier, CEDEX 05, 34095 Montpellier, FranceLaboratoire de Génie Chimique, Université de Toulouse, CNRS, INPT, UPS, 31432 Toulouse, FranceLaboratory of Pathogen-Host Interactions (LPHI), Université Montpellier, CEDEX 05, 34095 Montpellier, FranceThe opportunistic human pathogen <i>Pseudomonas aeruginosa</i> is responsible for a variety of acute infections and is a major cause of mortality in chronically infected cystic fibrosis patients. Due to increased resistance to antibiotics, new therapeutic strategies against <i>P. aeruginosa</i> are urgently needed. In this context, we aimed to develop a simple vertebrate animal model to rapidly assess in vivo drug efficacy against <i>P. aeruginosa</i>. Zebrafish are increasingly considered for modeling human infections caused by bacterial pathogens, which are commonly microinjected in embryos. In the present study, we established a novel protocol for zebrafish infection by <i>P. aeruginosa</i> based on bath immersion in 96-well plates of tail-injured embryos. The immersion method, followed by a 48-hour survey of embryo viability, was first validated to assess the virulence of <i>P. aeruginosa</i> wild-type PAO1 and a known attenuated mutant. We then validated its relevance for antipseudomonal drug testing by first using a clinically used antibiotic, ciprofloxacin. Secondly, we used a novel quorum sensing (QS) inhibitory molecule, <i>N</i>-(2-pyrimidyl)butanamide (C11), the activity of which had been validated in vitro but not previously tested in any animal model. A significant protective effect of C11 was observed on infected embryos, supporting the ability of C11 to attenuate in vivo <i>P. aeruginosa</i> pathogenicity. In conclusion, we present here a new and reliable method to compare the virulence of <i>P. aeruginosa</i> strains in vivo and to rapidly assess the efficacy of clinically relevant drugs against <i>P. aeruginosa</i>, including new antivirulence compounds.https://www.mdpi.com/2076-0817/10/4/401<i>Pseudomonas aeruginosa</i>zebrafishquorum sensing |
| spellingShingle | Pauline Nogaret Fatima El Garah Anne-Béatrice Blanc-Potard A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo Pathogens <i>Pseudomonas aeruginosa</i> zebrafish quorum sensing |
| title | A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo |
| title_full | A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo |
| title_fullStr | A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo |
| title_full_unstemmed | A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo |
| title_short | A Novel Infection Protocol in Zebrafish Embryo to Assess <i>Pseudomonas aeruginosa</i> Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo |
| title_sort | novel infection protocol in zebrafish embryo to assess i pseudomonas aeruginosa i virulence and validate efficacy of a quorum sensing inhibitor in vivo |
| topic | <i>Pseudomonas aeruginosa</i> zebrafish quorum sensing |
| url | https://www.mdpi.com/2076-0817/10/4/401 |
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