Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)

Candida albicans is both a commensal and a significant opportunistic fungal human pathogen, present in the digestive tract and the mouth as a part of the normal human microflora. Overgrowth of C. albicans can cause oral and vaginal candidiasis as well as a number of serious mucosal, and systemic lif...

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Main Author: Toutounji, Maggie Nazih (author)
Format: masterThesis
Published: 2018
Subjects:
Online Access:http://hdl.handle.net/10725/8627
https://doi.org/10.26756/th.2018.89
http://libraries.lau.edu.lb/research/laur/terms-of-use/thesis.php
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author Toutounji, Maggie Nazih
author_facet Toutounji, Maggie Nazih
author_role author
dc.creator.none.fl_str_mv Toutounji, Maggie Nazih
dc.date.none.fl_str_mv 2018-10-15T06:21:26Z
2018-10-15T06:21:26Z
2018
2018-10-15
2018-07-19
dc.identifier.none.fl_str_mv http://hdl.handle.net/10725/8627
https://doi.org/10.26756/th.2018.89
http://libraries.lau.edu.lb/research/laur/terms-of-use/thesis.php
dc.language.none.fl_str_mv en
dc.publisher.none.fl_str_mv Lebanese American University
dc.rights.*.fl_str_mv info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Lebanese American University -- Dissertations
Dissertations, Academic
Candida albicans
Drug resistance
Fungal cell walls -- Effect of drugs on
Hospitals -- Lebanon
dc.title.none.fl_str_mv Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
dc.type.none.fl_str_mv Thesis
info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/masterThesis
description Candida albicans is both a commensal and a significant opportunistic fungal human pathogen, present in the digestive tract and the mouth as a part of the normal human microflora. Overgrowth of C. albicans can cause oral and vaginal candidiasis as well as a number of serious mucosal, and systemic life threatening infections. The antifungal drug caspofungin of the echinocandin family is the latest generation of antifungal drugs to be developed. It functions by inhibiting glucan synthase thus weakening the fungal cell wall leading to death. Recently reports of resistance to caspofungin have been reported mainly through mutations in the FKS encoded subunits of glucan synthase at Hot spot 1 (amino acids 641 to 649, FSTLSLRDP) and hot spot 2 (amino acids 1357 to 1364, DWIRRYTL). Our study aimed at sequencing both hot spots from 16 C. albicans Lebanese hospital isolates resistant and sensitive to caspofungin to determine whether mutations in these hotspots are present, and whether such mutations also impart resistance to our isolates. In addition we wanted to determine any relationship between resistance and pathogenicity related attributes such as virulence, adhesion, filamentation, resistance to cell wall disrupting agents such as Congo red, biofilm formation, and cell wall chitin deposition. Five isolates were found to contain mutations with the mutations restricted to resistant strains. Within hotspot 1 substitution at positions S642, T643, L644, R647, and D648 were found, while within hotspot 2 substitutions at positions L1364, T1363, and R1360, W1358 and R1361 were identified. Interestingly some of the mutations found have not been previously documented. In addition strains that are resistant to caspofungin also showed increased resistance to Congo red but decreased biofilm formation and attenuated virulence in a mouse model of infection. Caspofungin sensitive strains showed decreased resistance to Congo red yet increased virulence and biofilm formation. All filamentous strains were adhesive but only some were virluent. Chitin content analysis showed that caspofungin resistant strains had elevated levels of chitin resulting in cell wall thickening that counters the effect of caspofungin, while sensitive strains showed decreased chitin content. Our results demonstrate an inverse correlation between resistance and virulence whereby resistance is due to thickening of the cell wall preventing the cell from gaining virulence attributes, while a more susceptible cell wall increases susceptibility to drugs but allows increased virulence.
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network_acronym_str LAURepo
network_name_str Lebanese American University repository
oai_identifier_str oai:laur.lau.edu.lb:10725/8627
publishDate 2018
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spelling Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)Toutounji, Maggie NazihLebanese American University -- DissertationsDissertations, AcademicCandida albicansDrug resistanceFungal cell walls -- Effect of drugs onHospitals -- LebanonCandida albicans is both a commensal and a significant opportunistic fungal human pathogen, present in the digestive tract and the mouth as a part of the normal human microflora. Overgrowth of C. albicans can cause oral and vaginal candidiasis as well as a number of serious mucosal, and systemic life threatening infections. The antifungal drug caspofungin of the echinocandin family is the latest generation of antifungal drugs to be developed. It functions by inhibiting glucan synthase thus weakening the fungal cell wall leading to death. Recently reports of resistance to caspofungin have been reported mainly through mutations in the FKS encoded subunits of glucan synthase at Hot spot 1 (amino acids 641 to 649, FSTLSLRDP) and hot spot 2 (amino acids 1357 to 1364, DWIRRYTL). Our study aimed at sequencing both hot spots from 16 C. albicans Lebanese hospital isolates resistant and sensitive to caspofungin to determine whether mutations in these hotspots are present, and whether such mutations also impart resistance to our isolates. In addition we wanted to determine any relationship between resistance and pathogenicity related attributes such as virulence, adhesion, filamentation, resistance to cell wall disrupting agents such as Congo red, biofilm formation, and cell wall chitin deposition. Five isolates were found to contain mutations with the mutations restricted to resistant strains. Within hotspot 1 substitution at positions S642, T643, L644, R647, and D648 were found, while within hotspot 2 substitutions at positions L1364, T1363, and R1360, W1358 and R1361 were identified. Interestingly some of the mutations found have not been previously documented. In addition strains that are resistant to caspofungin also showed increased resistance to Congo red but decreased biofilm formation and attenuated virulence in a mouse model of infection. Caspofungin sensitive strains showed decreased resistance to Congo red yet increased virulence and biofilm formation. All filamentous strains were adhesive but only some were virluent. Chitin content analysis showed that caspofungin resistant strains had elevated levels of chitin resulting in cell wall thickening that counters the effect of caspofungin, while sensitive strains showed decreased chitin content. Our results demonstrate an inverse correlation between resistance and virulence whereby resistance is due to thickening of the cell wall preventing the cell from gaining virulence attributes, while a more susceptible cell wall increases susceptibility to drugs but allows increased virulence.N/A1 hard copy: xi, 76 leaves; col. ill.; 30 cm. available at RNL.Bibliography : leaves 59-76.Lebanese American University2018-10-15T06:21:26Z2018-10-15T06:21:26Z20182018-10-152018-07-19Thesisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://hdl.handle.net/10725/8627https://doi.org/10.26756/th.2018.89http://libraries.lau.edu.lb/research/laur/terms-of-use/thesis.phpeninfo:eu-repo/semantics/openAccessoai:laur.lau.edu.lb:10725/86272021-03-19T10:45:18Z
spellingShingle Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
Toutounji, Maggie Nazih
Lebanese American University -- Dissertations
Dissertations, Academic
Candida albicans
Drug resistance
Fungal cell walls -- Effect of drugs on
Hospitals -- Lebanon
status_str publishedVersion
title Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
title_full Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
title_fullStr Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
title_full_unstemmed Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
title_short Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
title_sort Genotypic and phenotypic characterization of candida albicans Lebanese hospital isolates resistant and sensitive to caspofungin. (c2018)
topic Lebanese American University -- Dissertations
Dissertations, Academic
Candida albicans
Drug resistance
Fungal cell walls -- Effect of drugs on
Hospitals -- Lebanon
url http://hdl.handle.net/10725/8627
https://doi.org/10.26756/th.2018.89
http://libraries.lau.edu.lb/research/laur/terms-of-use/thesis.php