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Virulence Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/kvir20
Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa a
b
Sónia Gonçalves Pereira , Ana Cristina Rosa & Olga Cardoso a
a
Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal
b
Department of Mathematics, Faculty of Sciences and Technology, University of Coimbra, Coimbra, Portugal Accepted author version posted online: 07 May 2015.
Click for updates To cite this article: Sónia Gonçalves Pereira, Ana Cristina Rosa & Olga Cardoso (2015): Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa, Virulence, DOI: 10.1080/21505594.2015.1048958 To link to this article: http://dx.doi.org/10.1080/21505594.2015.1048958
Disclaimer: This is a version of an unedited manuscript that has been accepted for publication. As a service to authors and researchers we are providing this version of the accepted manuscript (AM). Copyediting, typesetting, and review of the resulting proof will be undertaken on this manuscript before final publication of the Version of Record (VoR). During production and pre-press, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal relate to this version also.
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa Sónia Gonçalves Pereira1, Ana Cristina Rosa2, Olga Cardoso1 1
Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal
2
Department of Mathematics, Faculty of Sciences and Technology, University of Coimbra, Coimbra,
Portugal
cr us an M ed pt ce Ac
Downloaded by [New York University] at 10:00 13 May 2015
ip t
Corresponding author: Sónia Gonçalves Pereira; e-mail: [email protected]
1
P. aeruginosa is the fifth most frequent pathogen worldwide, the second in nosocomial pneumonia, the third in urinary infections, the forth in post-surgical infections and the seventh pathogen responsible for sepsis.
1
It is also the major cause of mortality in cystic fibrosis patients and the most prevalent Gram-
negative multidrug-resistant bacteria in the airway of mechanically ventilated patients
2
and in pediatric
patients hospitalized in intensive care units (ICU). 1 Infections by P. aeruginosa are diverse and difficult
ip t
to treat due to its intrinsic ability to develop resistance under antibiotic pressure and to produce a variety of virulence factors (VF), like adhesins, proteases, phenazines and exotoxins. 3 Several studies report that
5,6
Some suggest
while others report enhanced resistance in isolates with high levels of
P. aeruginosa infections.
7,8
us
VF. 2,4 Also, literature indicates that inhibitors of specific VF may be good therapeutic alternatives to treat With present work, we intend to address possible associations between VF
an
and antibiotic resistance in P. aeruginosa, aiming to provide information that may be useful for the development of alternative therapies using VF inhibitors.
M
Seventy-six P. aeruginosa clinical isolates were randomly selected from sputum (43%), respiratory tract aspirates (21%), urine (20%), exudates (12%) and blood (4%) of inpatients and outpatients of a
ed
Portuguese central hospital, with consent from the patients. Antibiotic susceptibility tests were performed by disk diffusion method, using imipenem (IP), meropenem (MP), ceftazidime (CAZ), cefepime (FEP),
pt
aztreonam (AZT), piperacillin (PIP), amikacin (AMK) and ciprofloxacin (CIP). Results were interpreted according to CLSI recommendations, 9 with intermediate results considered resistant. MDR classification
ce
was performed according to Magiorakos et al. (2011). 10 Twelve virulence phenotypic characteristics were screened: swimming, swarming, twitching motility and production of rhamnolipids,
protease,
13
lipase, lecithinase
14
and pyocyanin.
15
Ac
Downloaded by [New York University] at 10:00 13 May 2015
these relations are antagonistic
2,4,5,6
cr
the presence or expression of virulence traits is related to resistance to antibiotics.
11
elastase,
12
Production of phospholipase C, pyoverdine and
biofilms were semi-quantified and classified as described elsewhere,
16,17,18
considering 4 classes of
production: absent, reduced, moderate and high. Screening for the genes flaG, orfF, pilA, pilB, associated
to motility; lecA, lecB, involved in lectin production; apr, lasA, lasB, encoding protease and elastases; phzH, phzM, phzS, phzI, phzII, from the biosynthetic pathway of phenazines; exoA, encoding exotoxinA and exoS, exoT, exoU and exoY, encoding type III secretory system effector proteins, was carried out as previously described. 3 exoA primers annealing temperature was 68ºC with expected amplified fragments of 368 bp. 19 Data were analyzed using SPSS® version 21.0 (IBM) software. Logistic regression (LR)
2
modeling of antibiotic resistance as function of VF presence was performed retaining the predictors statistically significant in Wald test (p
Virulence Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/kvir20
Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa a
b
Sónia Gonçalves Pereira , Ana Cristina Rosa & Olga Cardoso a
a
Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal
b
Department of Mathematics, Faculty of Sciences and Technology, University of Coimbra, Coimbra, Portugal Accepted author version posted online: 07 May 2015.
Click for updates To cite this article: Sónia Gonçalves Pereira, Ana Cristina Rosa & Olga Cardoso (2015): Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa, Virulence, DOI: 10.1080/21505594.2015.1048958 To link to this article: http://dx.doi.org/10.1080/21505594.2015.1048958
Disclaimer: This is a version of an unedited manuscript that has been accepted for publication. As a service to authors and researchers we are providing this version of the accepted manuscript (AM). Copyediting, typesetting, and review of the resulting proof will be undertaken on this manuscript before final publication of the Version of Record (VoR). During production and pre-press, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal relate to this version also.
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
Virulence factors as predictive tools for drug resistance in Pseudomonas aeruginosa Sónia Gonçalves Pereira1, Ana Cristina Rosa2, Olga Cardoso1 1
Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal
2
Department of Mathematics, Faculty of Sciences and Technology, University of Coimbra, Coimbra,
Portugal
cr us an M ed pt ce Ac
Downloaded by [New York University] at 10:00 13 May 2015
ip t
Corresponding author: Sónia Gonçalves Pereira; e-mail: [email protected]
1
P. aeruginosa is the fifth most frequent pathogen worldwide, the second in nosocomial pneumonia, the third in urinary infections, the forth in post-surgical infections and the seventh pathogen responsible for sepsis.
1
It is also the major cause of mortality in cystic fibrosis patients and the most prevalent Gram-
negative multidrug-resistant bacteria in the airway of mechanically ventilated patients
2
and in pediatric
patients hospitalized in intensive care units (ICU). 1 Infections by P. aeruginosa are diverse and difficult
ip t
to treat due to its intrinsic ability to develop resistance under antibiotic pressure and to produce a variety of virulence factors (VF), like adhesins, proteases, phenazines and exotoxins. 3 Several studies report that
5,6
Some suggest
while others report enhanced resistance in isolates with high levels of
P. aeruginosa infections.
7,8
us
VF. 2,4 Also, literature indicates that inhibitors of specific VF may be good therapeutic alternatives to treat With present work, we intend to address possible associations between VF
an
and antibiotic resistance in P. aeruginosa, aiming to provide information that may be useful for the development of alternative therapies using VF inhibitors.
M
Seventy-six P. aeruginosa clinical isolates were randomly selected from sputum (43%), respiratory tract aspirates (21%), urine (20%), exudates (12%) and blood (4%) of inpatients and outpatients of a
ed
Portuguese central hospital, with consent from the patients. Antibiotic susceptibility tests were performed by disk diffusion method, using imipenem (IP), meropenem (MP), ceftazidime (CAZ), cefepime (FEP),
pt
aztreonam (AZT), piperacillin (PIP), amikacin (AMK) and ciprofloxacin (CIP). Results were interpreted according to CLSI recommendations, 9 with intermediate results considered resistant. MDR classification
ce
was performed according to Magiorakos et al. (2011). 10 Twelve virulence phenotypic characteristics were screened: swimming, swarming, twitching motility and production of rhamnolipids,
protease,
13
lipase, lecithinase
14
and pyocyanin.
15
Ac
Downloaded by [New York University] at 10:00 13 May 2015
these relations are antagonistic
2,4,5,6
cr
the presence or expression of virulence traits is related to resistance to antibiotics.
11
elastase,
12
Production of phospholipase C, pyoverdine and
biofilms were semi-quantified and classified as described elsewhere,
16,17,18
considering 4 classes of
production: absent, reduced, moderate and high. Screening for the genes flaG, orfF, pilA, pilB, associated
to motility; lecA, lecB, involved in lectin production; apr, lasA, lasB, encoding protease and elastases; phzH, phzM, phzS, phzI, phzII, from the biosynthetic pathway of phenazines; exoA, encoding exotoxinA and exoS, exoT, exoU and exoY, encoding type III secretory system effector proteins, was carried out as previously described. 3 exoA primers annealing temperature was 68ºC with expected amplified fragments of 368 bp. 19 Data were analyzed using SPSS® version 21.0 (IBM) software. Logistic regression (LR)
2
modeling of antibiotic resistance as function of VF presence was performed retaining the predictors statistically significant in Wald test (p
