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Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples

Bacterial vaginosis (BV) is a common vaginal condition affecting reproductive-age women, especially in sub-Saharan Africa. With poor treatment outcomes, BV has been associated with pregnancy complications, pelvic inflammatory disease as well as acquisition and transmission of sexually transmitted di...

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Main Author: Masete, Kopano Valerie
Other Authors: Froissart, Rémy
Format: Thesis
Language:English
Published: Division of Medical Virology 2019
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access_status_str Open Access
author Masete, Kopano Valerie
author2 Froissart, Rémy
author_browse Froissart, Rémy
Masete, Kopano Valerie
author_facet Froissart, Rémy
Masete, Kopano Valerie
author_sort Masete, Kopano Valerie
collection Thesis
description Bacterial vaginosis (BV) is a common vaginal condition affecting reproductive-age women, especially in sub-Saharan Africa. With poor treatment outcomes, BV has been associated with pregnancy complications, pelvic inflammatory disease as well as acquisition and transmission of sexually transmitted diseases. While the etiology of BV is not well characterized, it is understood that Gardnerella vaginalis plays a critical role in BV by initiating the formation of the polymicrobial biofilm that characterizes BV and by degrading protective vaginal mucus through the release of sialidase. Recent evidence suggests that the G. vaginalis species is more heterogeneous that initially thought and that not all G. vaginalis may be involved BV. The aim of this study was thus to characterize the genotypic and phenotypic diversity of G. vaginalis isolates. This was achieved in vitro, using 109 G. vaginalis isolates that were previously purified from vaginal samples of 109 French women who were BV-positive (n = 75), BV-intermediate (n = 20) or BV-negative (n = 14), as diagnosed by Nugent scoring. To determine the genotypic diversity of G. vaginalis isolates, 90 isolates were successfully genotyped using their chaperonin-60 (cpn60) sequences, revealing the presence of four phylogenetic clades (subgroups A-D) made up of 13 subgroup A, 17 subgroup B, 58 subgroup C and 2 subgroup D isolates. To determine the phenotypic diversity of G. vaginalis isolates, sialidase activity, biofilm formation and susceptibility to antibiotics used to treat BV were measured. Sialidase activity was not detected in subgroup A and D isolates but was detected, at similar levels, in subgroup B and C isolates. Isolates from all subgroups of G. vaginalis could form similar amounts of biofilm. G. vaginalis isolates (n = 45) were largely resistant to metronidazole (71%), but sensitive to clindamycin (100%), moxifloxacin (91%) and augmentin (100%). The presence of prophages in G. vaginalis isolates was also investigated, revealing the presence of bacteriophage (phage)-like particles that could not be classified into any known phage families, whose phage status remains to be confirmed. In conclusion, G. vaginalis subgroup B and C isolates were the only ones that formed biofilm as well as had detectable sialidase activity suggesting that G. vaginalis subgroups B and C are most likely to be involved in BV. These results contribute to our knowledge of BV and could be useful in future studies that aim to design better treatment strategies for BV.
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institution University of Cape Town (South Africa)
language eng
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2019
publishDateRange 2019
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publisher Division of Medical Virology
publisherStr Division of Medical Virology
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spelling oai:open.uct.ac.za:11427/29274 Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples Masete, Kopano Valerie Froissart, Rémy Passmore, Jo-Ann Medical Virology Bacterial vaginosis (BV) is a common vaginal condition affecting reproductive-age women, especially in sub-Saharan Africa. With poor treatment outcomes, BV has been associated with pregnancy complications, pelvic inflammatory disease as well as acquisition and transmission of sexually transmitted diseases. While the etiology of BV is not well characterized, it is understood that Gardnerella vaginalis plays a critical role in BV by initiating the formation of the polymicrobial biofilm that characterizes BV and by degrading protective vaginal mucus through the release of sialidase. Recent evidence suggests that the G. vaginalis species is more heterogeneous that initially thought and that not all G. vaginalis may be involved BV. The aim of this study was thus to characterize the genotypic and phenotypic diversity of G. vaginalis isolates. This was achieved in vitro, using 109 G. vaginalis isolates that were previously purified from vaginal samples of 109 French women who were BV-positive (n = 75), BV-intermediate (n = 20) or BV-negative (n = 14), as diagnosed by Nugent scoring. To determine the genotypic diversity of G. vaginalis isolates, 90 isolates were successfully genotyped using their chaperonin-60 (cpn60) sequences, revealing the presence of four phylogenetic clades (subgroups A-D) made up of 13 subgroup A, 17 subgroup B, 58 subgroup C and 2 subgroup D isolates. To determine the phenotypic diversity of G. vaginalis isolates, sialidase activity, biofilm formation and susceptibility to antibiotics used to treat BV were measured. Sialidase activity was not detected in subgroup A and D isolates but was detected, at similar levels, in subgroup B and C isolates. Isolates from all subgroups of G. vaginalis could form similar amounts of biofilm. G. vaginalis isolates (n = 45) were largely resistant to metronidazole (71%), but sensitive to clindamycin (100%), moxifloxacin (91%) and augmentin (100%). The presence of prophages in G. vaginalis isolates was also investigated, revealing the presence of bacteriophage (phage)-like particles that could not be classified into any known phage families, whose phage status remains to be confirmed. In conclusion, G. vaginalis subgroup B and C isolates were the only ones that formed biofilm as well as had detectable sialidase activity suggesting that G. vaginalis subgroups B and C are most likely to be involved in BV. These results contribute to our knowledge of BV and could be useful in future studies that aim to design better treatment strategies for BV. 2019-02-05T06:32:01Z 2019-02-05T06:32:01Z 2018 2019-01-31T14:20:40Z Master Thesis Masters MSc http://hdl.handle.net/11427/29274 eng application/pdf Division of Medical Virology Faculty of Health Sciences University of Cape Town
spellingShingle Medical Virology
Masete, Kopano Valerie
Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
thesis_degree_str Master's
title Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
title_full Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
title_fullStr Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
title_full_unstemmed Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
title_short Characterizing the genotypic and phenotypic diversity of Gardnerella vaginalis from vaginal clinical samples
title_sort characterizing the genotypic and phenotypic diversity of gardnerella vaginalis from vaginal clinical samples
topic Medical Virology
url http://hdl.handle.net/11427/29274
work_keys_str_mv AT masetekopanovalerie characterizingthegenotypicandphenotypicdiversityofgardnerellavaginalisfromvaginalclinicalsamples