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Published in: BMC Infectious Diseases 1/2020

Open Access 01-12-2020 | Pneumonia | Research article

Characterization of the virulence of Pseudomonas aeruginosa strains causing ventilator-associated pneumonia

Authors: Beatriz Alonso, Laia Fernández-Barat, Enea Gino Di Domenico, Mercedes Marín, Emilia Cercenado, Irene Merino, Manuela de Pablos, Patricia Muñoz, María Guembe

Published in: BMC Infectious Diseases | Issue 1/2020

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Abstract

Background

The objective of this study was to evaluate the virulence of P. aeruginosa ventilator-associated pneumonia (VAP) strains (cases) in terms of biofilm production and other phenotypic and genotypic virulence factors compared to P. aeruginosa strains isolated from other infections (controls).

Methods

Biofilm production was tested to assess biomass production and metabolic activity using crystal violet binding assay and XTT assay, respectively. Pigment production (pyocyanin and pyoverdine) was evaluated using cetrimide agar. Virulence genes were detected by conventional multiplex PCR and virulence was tested in an in vivo model in Galleria mellonella larvae.

Results

We did not find statistically significant differences between VAP and no-VAP strains (p > 0.05) regarding biofilm production. VAP strains had no production of pyocyanin after 24 h of incubation (p = 0.023). The distribution of virulence genes between both groups were similar (p > 0.05). VAP strains were less virulent than non-VAP strains in an in vivo model of G. mellonella (p < 0.001).

Conclusion

The virulence of VAP-Pseudomonas aeruginosa does not depend on biofilm formation, production of pyoverdine or the presence of some virulence genes compared to P. aeruginosa isolated from non-invasive locations. However, VAP strains showed attenuated virulence compared to non-VAP strains in an in vivo model of G. mellonella.
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Literature
3.
go back to reference Kalil AC, Metersky ML, Klompas M, Muscedere J, Sweeney DA, Palmer LB, et al. Management of Adults with Hospital-acquired and Ventilator-associated Pneumonia: 2016 clinical practice guidelines by the Infectious Diseases Society of America and the American Thoracic Society. Clin Infect Dis. 2016;63(5):e61–e111.CrossRefPubMedPubMedCentral Kalil AC, Metersky ML, Klompas M, Muscedere J, Sweeney DA, Palmer LB, et al. Management of Adults with Hospital-acquired and Ventilator-associated Pneumonia: 2016 clinical practice guidelines by the Infectious Diseases Society of America and the American Thoracic Society. Clin Infect Dis. 2016;63(5):e61–e111.CrossRefPubMedPubMedCentral
4.
go back to reference Sarda C, Fazal F, Rello J. Management of ventilator-associated pneumonia (VAP) caused by resistant gram-negative bacteria: which is the best strategy to treat? Expert Rev Respir Med. 2019;13(8):787–98.CrossRefPubMed Sarda C, Fazal F, Rello J. Management of ventilator-associated pneumonia (VAP) caused by resistant gram-negative bacteria: which is the best strategy to treat? Expert Rev Respir Med. 2019;13(8):787–98.CrossRefPubMed
5.
go back to reference Maurice NM, Bedi B, Sadikot RT. Pseudomonas aeruginosa biofilms: host response and clinical implications in lung infections. Am J Respir Cell Mol Biol. 2018;58(4):428–39.CrossRefPubMedPubMedCentral Maurice NM, Bedi B, Sadikot RT. Pseudomonas aeruginosa biofilms: host response and clinical implications in lung infections. Am J Respir Cell Mol Biol. 2018;58(4):428–39.CrossRefPubMedPubMedCentral
6.
go back to reference Fernandez-Barat L, Torres A. Biofilms in ventilator-associated pneumonia. Future Microbiol. 2016;11:1599–610. Fernandez-Barat L, Torres A. Biofilms in ventilator-associated pneumonia. Future Microbiol. 2016;11:1599–610.
7.
go back to reference Berra L, Schmidt U, Wiener-Kronish J. Relationship between virulence factors and outcome of ventilator-associated pneumonia related to Pseudomonas aeruginosa. Curr Respir Med Rev. 2010;6(1):19–25.CrossRef Berra L, Schmidt U, Wiener-Kronish J. Relationship between virulence factors and outcome of ventilator-associated pneumonia related to Pseudomonas aeruginosa. Curr Respir Med Rev. 2010;6(1):19–25.CrossRef
8.
go back to reference Gellatly SL, Hancock RE. Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathog Disease. 2013;67(3):159–73.CrossRef Gellatly SL, Hancock RE. Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathog Disease. 2013;67(3):159–73.CrossRef
9.
go back to reference Alonso B, Lucio J, Perez-Granda MJ, Cruces R, Sanchez-Carrillo C, Bouza E, et al. Does biomass production correlate with metabolic activity in Staphylococcus aureus? J Microbiol Methods. 2016;131:110–12. Alonso B, Lucio J, Perez-Granda MJ, Cruces R, Sanchez-Carrillo C, Bouza E, et al. Does biomass production correlate with metabolic activity in Staphylococcus aureus? J Microbiol Methods. 2016;131:110–12.
10.
go back to reference Stepanovic S, Vukovic D, Hola V, Di Bonaventura G, Djukic S, Cirkovic I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007;115(8):891–9.CrossRefPubMed Stepanovic S, Vukovic D, Hola V, Di Bonaventura G, Djukic S, Cirkovic I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007;115(8):891–9.CrossRefPubMed
11.
go back to reference Fazeli N, Momtaz H. Virulence gene profiles of multidrug-resistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red Crescent Med J. 2014;16(10):e15722.CrossRefPubMedPubMedCentral Fazeli N, Momtaz H. Virulence gene profiles of multidrug-resistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red Crescent Med J. 2014;16(10):e15722.CrossRefPubMedPubMedCentral
12.
go back to reference Gholami A, Majidpour A, Talebi-Taher M, Boustanshenas M, Adabi M. PCR-based assay for the rapid and precise distinction of Pseudomonas aeruginosa from other Pseudomonas species recovered from burns patients. J Prev Med Hyg. 2016;57(2):E81–5.PubMedPubMedCentral Gholami A, Majidpour A, Talebi-Taher M, Boustanshenas M, Adabi M. PCR-based assay for the rapid and precise distinction of Pseudomonas aeruginosa from other Pseudomonas species recovered from burns patients. J Prev Med Hyg. 2016;57(2):E81–5.PubMedPubMedCentral
13.
go back to reference Sabharwal N, Dhall S, Chhibber S, Harjai K. Molecular detection of virulence genes as markers in Pseudomonas aeruginosa isolated from urinary tract infections. Int J Mol Epidemiol Genet. 2014;5(3):125–34.PubMedPubMedCentral Sabharwal N, Dhall S, Chhibber S, Harjai K. Molecular detection of virulence genes as markers in Pseudomonas aeruginosa isolated from urinary tract infections. Int J Mol Epidemiol Genet. 2014;5(3):125–34.PubMedPubMedCentral
14.
go back to reference Marcos-Zambrano LJ, Puig-Asensio M, Perez-Garcia F, Escribano P, Sanchez-Carrillo C, Zaragoza O, et al. Candida guilliermondii complex is characterized by high antifungal resistance but low mortality in 22 cases of candidemia. Antimicrob Agents Chemother. 2017;61(7):e00099–17. Marcos-Zambrano LJ, Puig-Asensio M, Perez-Garcia F, Escribano P, Sanchez-Carrillo C, Zaragoza O, et al. Candida guilliermondii complex is characterized by high antifungal resistance but low mortality in 22 cases of candidemia. Antimicrob Agents Chemother. 2017;61(7):e00099–17.
15.
go back to reference Weiss E, Essaied W, Adrie C, Zahar JR, Timsit JF. Treatment of severe hospital-acquired and ventilator-associated pneumonia: a systematic review of inclusion and judgment criteria used in randomized controlled trials. Crit Care. 2017;21(1):162.CrossRefPubMedPubMedCentral Weiss E, Essaied W, Adrie C, Zahar JR, Timsit JF. Treatment of severe hospital-acquired and ventilator-associated pneumonia: a systematic review of inclusion and judgment criteria used in randomized controlled trials. Crit Care. 2017;21(1):162.CrossRefPubMedPubMedCentral
16.
go back to reference Landelle C, Nocquet Boyer V, Abbas M, Genevois E, Abidi N, Naimo S, et al. Impact of a multifaceted prevention program on ventilator-associated pneumonia including selective oropharyngeal decontamination. Intensive Care Med. 2018;44(11):1777–86.CrossRefPubMedPubMedCentral Landelle C, Nocquet Boyer V, Abbas M, Genevois E, Abidi N, Naimo S, et al. Impact of a multifaceted prevention program on ventilator-associated pneumonia including selective oropharyngeal decontamination. Intensive Care Med. 2018;44(11):1777–86.CrossRefPubMedPubMedCentral
17.
go back to reference Tokmaji G, Vermeulen H, Muller MC, Kwakman PH, Schultz MJ, Zaat SA. Silver-coated endotracheal tubes for prevention of ventilator-associated pneumonia in critically ill patients. Cochrane Database Syst Rev. 2015(8):Cd009201. Tokmaji G, Vermeulen H, Muller MC, Kwakman PH, Schultz MJ, Zaat SA. Silver-coated endotracheal tubes for prevention of ventilator-associated pneumonia in critically ill patients. Cochrane Database Syst Rev. 2015(8):Cd009201.
18.
go back to reference Sadikot RT, Blackwell TS, Christman JW, Prince AS. Pathogen-host interactions in Pseudomonas aeruginosa pneumonia. Am J Respir Crit Care Med. 2005;171(11):1209–23.CrossRefPubMedPubMedCentral Sadikot RT, Blackwell TS, Christman JW, Prince AS. Pathogen-host interactions in Pseudomonas aeruginosa pneumonia. Am J Respir Crit Care Med. 2005;171(11):1209–23.CrossRefPubMedPubMedCentral
19.
go back to reference Kohler T, Guanella R, Carlet J, van Delden C. Quorum sensing-dependent virulence during Pseudomonas aeruginosa colonisation and pneumonia in mechanically ventilated patients. Thorax. 2010;65(8):703–10.CrossRefPubMed Kohler T, Guanella R, Carlet J, van Delden C. Quorum sensing-dependent virulence during Pseudomonas aeruginosa colonisation and pneumonia in mechanically ventilated patients. Thorax. 2010;65(8):703–10.CrossRefPubMed
20.
go back to reference Rybtke M, Hultqvist LD, Givskov M, Tolker-Nielsen T. Pseudomonas aeruginosa biofilm infections: community structure, antimicrobial tolerance and immune response. J Mol Biol. 2015;427(23):3628–45.CrossRefPubMed Rybtke M, Hultqvist LD, Givskov M, Tolker-Nielsen T. Pseudomonas aeruginosa biofilm infections: community structure, antimicrobial tolerance and immune response. J Mol Biol. 2015;427(23):3628–45.CrossRefPubMed
21.
go back to reference Lee K, Yoon SS. Pseudomonas aeruginosa biofilm, a programmed bacterial life for fitness. J Microbiol Biotechnol. 2017;27(6):1053–64.CrossRefPubMed Lee K, Yoon SS. Pseudomonas aeruginosa biofilm, a programmed bacterial life for fitness. J Microbiol Biotechnol. 2017;27(6):1053–64.CrossRefPubMed
22.
go back to reference Rada B, Leto TL. Pyocyanin effects on respiratory epithelium: relevance in Pseudomonas aeruginosa airway infections. Trends Microbiol. 2013;21(2):73–81.CrossRefPubMed Rada B, Leto TL. Pyocyanin effects on respiratory epithelium: relevance in Pseudomonas aeruginosa airway infections. Trends Microbiol. 2013;21(2):73–81.CrossRefPubMed
23.
go back to reference Lau GW, Hassett DJ, Ran H, Kong F. The role of pyocyanin in Pseudomonas aeruginosa infection. Trends Mol Med. 2004;10(12):599–606.CrossRefPubMed Lau GW, Hassett DJ, Ran H, Kong F. The role of pyocyanin in Pseudomonas aeruginosa infection. Trends Mol Med. 2004;10(12):599–606.CrossRefPubMed
24.
go back to reference Stacey SD, Pritchett CL. Pseudomonas aeruginosa AlgU contributes to posttranscriptional activity by increasing rsmA expression in a mucA22 strain. J Bacteriol. 2016;198(13):1812–26.CrossRefPubMedPubMedCentral Stacey SD, Pritchett CL. Pseudomonas aeruginosa AlgU contributes to posttranscriptional activity by increasing rsmA expression in a mucA22 strain. J Bacteriol. 2016;198(13):1812–26.CrossRefPubMedPubMedCentral
25.
go back to reference Azam MW, Khan AU. Updates on the pathogenicity status of Pseudomonas aeruginosa. Drug Discov Today. 2019;24(1):350–59. Azam MW, Khan AU. Updates on the pathogenicity status of Pseudomonas aeruginosa. Drug Discov Today. 2019;24(1):350–59.
26.
go back to reference Sawa T. The molecular mechanism of acute lung injury caused by Pseudomonas aeruginosa: from bacterial pathogenesis to host response. J Intensive Care. 2014;2(1):10.CrossRefPubMedPubMedCentral Sawa T. The molecular mechanism of acute lung injury caused by Pseudomonas aeruginosa: from bacterial pathogenesis to host response. J Intensive Care. 2014;2(1):10.CrossRefPubMedPubMedCentral
27.
go back to reference Wolfgang MC, Kulasekara BR, Liang X, Boyd D, Wu K, Yang Q, et al. Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2003;100(14):8484–9.CrossRefPubMedPubMedCentral Wolfgang MC, Kulasekara BR, Liang X, Boyd D, Wu K, Yang Q, et al. Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2003;100(14):8484–9.CrossRefPubMedPubMedCentral
28.
go back to reference Tsai CJ, Loh JM, Proft T. Galleria mellonella infection models for the study of bacterial diseases and for antimicrobial drug testing. Virulence. 2016;7(3):214–29.CrossRefPubMedPubMedCentral Tsai CJ, Loh JM, Proft T. Galleria mellonella infection models for the study of bacterial diseases and for antimicrobial drug testing. Virulence. 2016;7(3):214–29.CrossRefPubMedPubMedCentral
29.
go back to reference Wang K, Chen YQ, Salido MM, Kohli GS, Kong JL, Liang HJ, et al. The rapid in vivo evolution of Pseudomonas aeruginosa in ventilator-associated pneumonia patients leads to attenuated virulence. Open Biol. 2017;7(9):170029. Wang K, Chen YQ, Salido MM, Kohli GS, Kong JL, Liang HJ, et al. The rapid in vivo evolution of Pseudomonas aeruginosa in ventilator-associated pneumonia patients leads to attenuated virulence. Open Biol. 2017;7(9):170029.
30.
go back to reference Kohler T, Buckling A, van Delden C. Cooperation and virulence of clinical Pseudomonas aeruginosa populations. Proc Natl Acad Sci U S A. 2009;106(15):6339–44.CrossRefPubMedPubMedCentral Kohler T, Buckling A, van Delden C. Cooperation and virulence of clinical Pseudomonas aeruginosa populations. Proc Natl Acad Sci U S A. 2009;106(15):6339–44.CrossRefPubMedPubMedCentral
31.
go back to reference De Winter FHR, s Jongers B Bielen K, Mancuso D, Timbermont L, Lammens C, et al. Mechanical ventilation impairs IL-17 cytokine family expression in ventilator-associated pneumonia. 2019;20(20):5072. De Winter FHR, s Jongers B Bielen K, Mancuso D, Timbermont L, Lammens C, et al. Mechanical ventilation impairs IL-17 cytokine family expression in ventilator-associated pneumonia. 2019;20(20):5072.
Metadata
Title
Characterization of the virulence of Pseudomonas aeruginosa strains causing ventilator-associated pneumonia
Authors
Beatriz Alonso
Laia Fernández-Barat
Enea Gino Di Domenico
Mercedes Marín
Emilia Cercenado
Irene Merino
Manuela de Pablos
Patricia Muñoz
María Guembe
Publication date
01-12-2020
Publisher
BioMed Central
Published in
BMC Infectious Diseases / Issue 1/2020
Electronic ISSN: 1471-2334
DOI
https://doi.org/10.1186/s12879-020-05534-1

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