Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal

Detalhes bibliográficos
Autor(a) principal: Almeida, Geone Pimentel dos Santos Bulhões de
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal de Alagoas (UFAL)
Texto Completo: http://www.repositorio.ufal.br/handle/riufal/7315
Resumo: Vulvovaginal candidiasis (VVC) is a fungal infection classified as a sexually transmitted disease according to WHO, caused by yeast of the genus Candida. In Brazil, it is the second cause of infections among women of childbearing age and sexually active. The therapeutic treatment available for VVC treatment is restricted to polyene and azole drugs and resistance to the recommended antifungal agents (ketoconazole / CET, fluconazole / FLU, itraconazole / ITR and nystatin / NIS) has been reported more frequently in Candida albicans , the main associated species, but also in Candida non-albicans species (eg C.glabrata, C. krusei and C. parapsilosis complex). Propolis is a resinous compound produced by bees Apis mellifera L. and is known to possess a variety of biological activities, and Brazilian red propolis (BRP) has components such as flavonoids that cause structural and functional damage against microorganisms. The objective of this study was to evaluate the in vitro antifungal activity of PVA (ethanol extract and fractions) against different Candida species, focusing on clinical isolates of VVC. Five strains of ATCC reference (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei) and 15 clinical isolates (3 of each species) were cultured from patients with VVC from Maceió / Alagoas on Sabouraud agar Dextrose and CHROMagar® Candida separately until 48 hours at 37° C. Suspensions of individual Candida cultures were adjusted to 0.5 McFarland standard in 0.9% NaCl solution for antimicrobial susceptibility testing (TSA) by disc diffusion and microdilution methods, as CLSI for CET, ITR, FLU , NIS, as well as EE and its fractions. The JASP (version 0.8.3.1) and IBM®SPSS (version 23.0) programs were adopted in the statistical analyzes. It was observed that in the TSA by disk diffusion and microdilution with both commercial antifungal and EE and fractions showed varied susceptibility profiles for the analyzed species, mainly among the clinical isolates when compared to the reference strains. By the disc-diffusion method all ATCC strains were susceptible to CET and NIS, but there was resistance to ITR and FLU mainly in C. glabrata and C. krusei, including ITR / FLU co-resistance pattern confirmed in 9 isolates. Among all the clinical isolates, resistance to at least 2 antifungal agents was tested, being 8 resistant to CET / ITR / FLU, drugs indicated by RENAME (SUS) for VVC treatment. The microdilution tests confirmed that CET and NIS were more effective, whereas FLU and ITR had less activity in the reference strains and some clinical isolates. The microdilution method with PVA showed MICs for the fractions ranging from 125 to 250 μg / mL, being found MIC of 500 μg / m only for C. parapsilosis and C. krusei in relation to the hexanolic and ethanolic fractions, respectively. The ethanolic fraction of PVA was effective against a larger number of strains tested, but the acetanolic fraction acted with lower MIC, probably because of more flavonoids, whereas the chloroform fraction had homogenous MIC among the reference strains (250 μg / mL) . TSA by both methods indicated 9 clinical isolates with ITR / FLU co-resistance, and PVA was effective with low MIC for 6 isolates with R-ITR / FLU standard. Disk-diffusion and microdilution tests were concordant in most of the strains and isolates tested. These results provide new data on the Brazilian red propolis of Alagoas in relation to the quality of flavonoids and confirmed the anti-Candida activity of different fractions for several species frequently involved in vulvovaginal candidiasis, creating possibilities for the use of PVA as a bioactive with important pharmacological potential in formulations of vaginal use.
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spelling Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase VulvovaginalCandidíase vulvovaginalPrópole - AlagoasAntifúngicosVulvovaginal candidiasisred propolisantifungalsCNPQ::CIENCIAS DA SAUDE::FARMACIAVulvovaginal candidiasis (VVC) is a fungal infection classified as a sexually transmitted disease according to WHO, caused by yeast of the genus Candida. In Brazil, it is the second cause of infections among women of childbearing age and sexually active. The therapeutic treatment available for VVC treatment is restricted to polyene and azole drugs and resistance to the recommended antifungal agents (ketoconazole / CET, fluconazole / FLU, itraconazole / ITR and nystatin / NIS) has been reported more frequently in Candida albicans , the main associated species, but also in Candida non-albicans species (eg C.glabrata, C. krusei and C. parapsilosis complex). Propolis is a resinous compound produced by bees Apis mellifera L. and is known to possess a variety of biological activities, and Brazilian red propolis (BRP) has components such as flavonoids that cause structural and functional damage against microorganisms. The objective of this study was to evaluate the in vitro antifungal activity of PVA (ethanol extract and fractions) against different Candida species, focusing on clinical isolates of VVC. Five strains of ATCC reference (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei) and 15 clinical isolates (3 of each species) were cultured from patients with VVC from Maceió / Alagoas on Sabouraud agar Dextrose and CHROMagar® Candida separately until 48 hours at 37° C. Suspensions of individual Candida cultures were adjusted to 0.5 McFarland standard in 0.9% NaCl solution for antimicrobial susceptibility testing (TSA) by disc diffusion and microdilution methods, as CLSI for CET, ITR, FLU , NIS, as well as EE and its fractions. The JASP (version 0.8.3.1) and IBM®SPSS (version 23.0) programs were adopted in the statistical analyzes. It was observed that in the TSA by disk diffusion and microdilution with both commercial antifungal and EE and fractions showed varied susceptibility profiles for the analyzed species, mainly among the clinical isolates when compared to the reference strains. By the disc-diffusion method all ATCC strains were susceptible to CET and NIS, but there was resistance to ITR and FLU mainly in C. glabrata and C. krusei, including ITR / FLU co-resistance pattern confirmed in 9 isolates. Among all the clinical isolates, resistance to at least 2 antifungal agents was tested, being 8 resistant to CET / ITR / FLU, drugs indicated by RENAME (SUS) for VVC treatment. The microdilution tests confirmed that CET and NIS were more effective, whereas FLU and ITR had less activity in the reference strains and some clinical isolates. The microdilution method with PVA showed MICs for the fractions ranging from 125 to 250 μg / mL, being found MIC of 500 μg / m only for C. parapsilosis and C. krusei in relation to the hexanolic and ethanolic fractions, respectively. The ethanolic fraction of PVA was effective against a larger number of strains tested, but the acetanolic fraction acted with lower MIC, probably because of more flavonoids, whereas the chloroform fraction had homogenous MIC among the reference strains (250 μg / mL) . TSA by both methods indicated 9 clinical isolates with ITR / FLU co-resistance, and PVA was effective with low MIC for 6 isolates with R-ITR / FLU standard. Disk-diffusion and microdilution tests were concordant in most of the strains and isolates tested. These results provide new data on the Brazilian red propolis of Alagoas in relation to the quality of flavonoids and confirmed the anti-Candida activity of different fractions for several species frequently involved in vulvovaginal candidiasis, creating possibilities for the use of PVA as a bioactive with important pharmacological potential in formulations of vaginal use.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorA candidíase vulvovaginal (CVV) é considera uma doença sexualmente transmissível segundo a OMS, causada por levedura do gênero Candida. No Brasil, é a segunda causa de infecções entre mulheres em idade fértil e sexualmente ativas. O tratamento terapêutico disponível para o tratamento com CVV é restrito a fármacos poliênicos e azólicos, e a resistência à agentes antifúngicos recomendados (cetoconazol/CET, fluconazol/FLU, itraconazol/ITR e nistatina/NIS) tem sido relatada com mais frequência em Candida albicans, a principal espécie associada, mas também em espécies de Candida não-albicans (p. ex, C.glabrata, C. krusei e do complexo de C. parapsilosis). A própolis é um composto resinoso produzido por abelhas Apis mellifera L. e é conhecido por possuir uma variedade de atividades biológicas, e a própolis vermelha de Alagoas (PVA) tem componentes como flavonóides que causam danos estruturais e funcionais contra microorganismos. O objetivo deste estudo foi avaliar a atividade antifúngica in vitro de PVA (extrato de etanol e frações) contra diferentes espécies de Candida, focalizando isolados clínicos de CVV. Foram cultivadas cinco cepas de referência ATCC (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis e C. krusei) e 15 isolados clínicos (3 de cada espécie citada) de pacientes com CVV de Maceió/Alagoas em ágar Sabouraud Dextrose e CHROMagar®Candida separadamente, até 48h a 37°C. As suspensões de cada cultura de Candida foram ajustadas para 0,5 na escala McFarland em solução NaCl 0,9% para os testes de susceptibilidade a antimicrobianos (TSA) pelos métodos de difusão de disco e microdiluição, conforme CLSI para CET, ITR, FLU, NIS, bem como EE e suas frações. Os programas JASP (versão 0.8.3.1) e IBM®SPSS (versão 23.0) foram adotados nas análises estatísticas. Observouse que no TSA por disco-difusão e microdiluição tanto com antifúngicos comerciais como com EE e frações mostraram perfis de susceptibilidade variados para as espécies analisadas, principalmente entre os isolados clínicos quando comparados às cepas de referência. Pelo método disco-difusão todas as cepas ATCC foram susceptíveis a CET e NIS, mas houve resistência ao ITR e FLU principalmente em C. glabrata e C. krusei, inclusive padrão de coresistência ITR/FLU confirmado em 9 isolados. Entre todos os isolados clínicos houve resistência a pelo menos 2 antifúngicos testados, sendo 8 resistentes a CET/ITR/FLU, drogas indicadas pelo RENAME (SUS) para tratamento de CVV. Os testes de microdiluição confirmaram que CET e NIS se mostraram mais efetivos, enquanto FLU e ITR tiveram menor atividade nas cepas de referência e alguns isolados clínicos. O método de microdiluição com a PVA mostraram CIM para as frações variando de 125 a 250 μg/mL, sendo constatado CIM de 500 μg/m apenas para C. parapsilosis e C. krusei em relação às frações hexanólica e etanólico, respectivamente. A fração etanólica de PVA foi eficaz contra uma maior quantidade de cepas testadas, mas a fração acetanólica agiu com CIM mais baixo, provavelmente por que mais flavonoides, enquanto que a fração clorofórmica teve CIM homogêneo entre as cepas de referência (250 μg/mL). TSA por ambos os métodos indicou 9 isolados clínicos com coresistência à ITR/FLU, e PVA foi efetiva com baixo CIM para 6 isolados com padrão RITR/ FLU. Os testes de disco-difusão e microdiluição foram concordantes na maior parte das cepas e isolados testados. Esses resultados trazem novos dados sobre a própolis vermelha brasileira de Alagoas em relação à qualidade de flavonoides e confirmou atividade anti-Candida de diferentes frações para várias espécies frequentemente envolvidas em candidíase vulvovaginal, criando possibilidades para o uso de PVA como bioativo com importante potencial farmacológico em formulações de uso vaginal.Universidade Federal de AlagoasBrasilPrograma de Pós-Graduação em Ciências FarmacêuticasUFALReys, José Rui Machadohttp://lattes.cnpq.br/5637387651693816Maranhão, Fernanda Cristina de Albuquerquehttp://lattes.cnpq.br/9154361722501415Moura, Maria Aline Barros Fidelis dehttp://lattes.cnpq.br/8554388756291432Nascimento, Ticiano Gomes dohttp://lattes.cnpq.br/6296388037177344Silva Filho, Eurípedes Alves dahttp://lattes.cnpq.br/8114140674559096Almeida, Geone Pimentel dos Santos Bulhões de2020-11-12T02:15:35Z2020-11-012020-11-12T02:15:35Z2017-12-07info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfALMEIDA, Geone Pimentel dos Santos Bulhões de. Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal. 2017. 89 f. Dissertação (Mestrado em Ciências Farmacêuticas) – Instituto de Ciências Farmacêuticas, Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal de Alagoas, Maceió, 2020.http://www.repositorio.ufal.br/handle/riufal/7315porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal de Alagoas (UFAL)instname:Universidade Federal de Alagoas (UFAL)instacron:UFAL2020-11-12T02:17:45Zoai:www.repositorio.ufal.br:riufal/7315Repositório InstitucionalPUBhttp://www.repositorio.ufal.br/oai/requestri@sibi.ufal.bropendoar:2020-11-12T02:17:45Repositório Institucional da Universidade Federal de Alagoas (UFAL) - Universidade Federal de Alagoas (UFAL)false
dc.title.none.fl_str_mv Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
title Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
spellingShingle Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
Almeida, Geone Pimentel dos Santos Bulhões de
Candidíase vulvovaginal
Própole - Alagoas
Antifúngicos
Vulvovaginal candidiasis
red propolis
antifungals
CNPQ::CIENCIAS DA SAUDE::FARMACIA
title_short Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
title_full Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
title_fullStr Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
title_full_unstemmed Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
title_sort Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal
author Almeida, Geone Pimentel dos Santos Bulhões de
author_facet Almeida, Geone Pimentel dos Santos Bulhões de
author_role author
dc.contributor.none.fl_str_mv Reys, José Rui Machado
http://lattes.cnpq.br/5637387651693816
Maranhão, Fernanda Cristina de Albuquerque
http://lattes.cnpq.br/9154361722501415
Moura, Maria Aline Barros Fidelis de
http://lattes.cnpq.br/8554388756291432
Nascimento, Ticiano Gomes do
http://lattes.cnpq.br/6296388037177344
Silva Filho, Eurípedes Alves da
http://lattes.cnpq.br/8114140674559096
dc.contributor.author.fl_str_mv Almeida, Geone Pimentel dos Santos Bulhões de
dc.subject.por.fl_str_mv Candidíase vulvovaginal
Própole - Alagoas
Antifúngicos
Vulvovaginal candidiasis
red propolis
antifungals
CNPQ::CIENCIAS DA SAUDE::FARMACIA
topic Candidíase vulvovaginal
Própole - Alagoas
Antifúngicos
Vulvovaginal candidiasis
red propolis
antifungals
CNPQ::CIENCIAS DA SAUDE::FARMACIA
description Vulvovaginal candidiasis (VVC) is a fungal infection classified as a sexually transmitted disease according to WHO, caused by yeast of the genus Candida. In Brazil, it is the second cause of infections among women of childbearing age and sexually active. The therapeutic treatment available for VVC treatment is restricted to polyene and azole drugs and resistance to the recommended antifungal agents (ketoconazole / CET, fluconazole / FLU, itraconazole / ITR and nystatin / NIS) has been reported more frequently in Candida albicans , the main associated species, but also in Candida non-albicans species (eg C.glabrata, C. krusei and C. parapsilosis complex). Propolis is a resinous compound produced by bees Apis mellifera L. and is known to possess a variety of biological activities, and Brazilian red propolis (BRP) has components such as flavonoids that cause structural and functional damage against microorganisms. The objective of this study was to evaluate the in vitro antifungal activity of PVA (ethanol extract and fractions) against different Candida species, focusing on clinical isolates of VVC. Five strains of ATCC reference (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei) and 15 clinical isolates (3 of each species) were cultured from patients with VVC from Maceió / Alagoas on Sabouraud agar Dextrose and CHROMagar® Candida separately until 48 hours at 37° C. Suspensions of individual Candida cultures were adjusted to 0.5 McFarland standard in 0.9% NaCl solution for antimicrobial susceptibility testing (TSA) by disc diffusion and microdilution methods, as CLSI for CET, ITR, FLU , NIS, as well as EE and its fractions. The JASP (version 0.8.3.1) and IBM®SPSS (version 23.0) programs were adopted in the statistical analyzes. It was observed that in the TSA by disk diffusion and microdilution with both commercial antifungal and EE and fractions showed varied susceptibility profiles for the analyzed species, mainly among the clinical isolates when compared to the reference strains. By the disc-diffusion method all ATCC strains were susceptible to CET and NIS, but there was resistance to ITR and FLU mainly in C. glabrata and C. krusei, including ITR / FLU co-resistance pattern confirmed in 9 isolates. Among all the clinical isolates, resistance to at least 2 antifungal agents was tested, being 8 resistant to CET / ITR / FLU, drugs indicated by RENAME (SUS) for VVC treatment. The microdilution tests confirmed that CET and NIS were more effective, whereas FLU and ITR had less activity in the reference strains and some clinical isolates. The microdilution method with PVA showed MICs for the fractions ranging from 125 to 250 μg / mL, being found MIC of 500 μg / m only for C. parapsilosis and C. krusei in relation to the hexanolic and ethanolic fractions, respectively. The ethanolic fraction of PVA was effective against a larger number of strains tested, but the acetanolic fraction acted with lower MIC, probably because of more flavonoids, whereas the chloroform fraction had homogenous MIC among the reference strains (250 μg / mL) . TSA by both methods indicated 9 clinical isolates with ITR / FLU co-resistance, and PVA was effective with low MIC for 6 isolates with R-ITR / FLU standard. Disk-diffusion and microdilution tests were concordant in most of the strains and isolates tested. These results provide new data on the Brazilian red propolis of Alagoas in relation to the quality of flavonoids and confirmed the anti-Candida activity of different fractions for several species frequently involved in vulvovaginal candidiasis, creating possibilities for the use of PVA as a bioactive with important pharmacological potential in formulations of vaginal use.
publishDate 2017
dc.date.none.fl_str_mv 2017-12-07
2020-11-12T02:15:35Z
2020-11-01
2020-11-12T02:15:35Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv ALMEIDA, Geone Pimentel dos Santos Bulhões de. Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal. 2017. 89 f. Dissertação (Mestrado em Ciências Farmacêuticas) – Instituto de Ciências Farmacêuticas, Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal de Alagoas, Maceió, 2020.
http://www.repositorio.ufal.br/handle/riufal/7315
identifier_str_mv ALMEIDA, Geone Pimentel dos Santos Bulhões de. Atividade antifúngica in vitro de Extrato da Própolis Vermelha de Alagoas em Candida spp. isoladas de pacientes com Candidíase Vulvovaginal. 2017. 89 f. Dissertação (Mestrado em Ciências Farmacêuticas) – Instituto de Ciências Farmacêuticas, Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal de Alagoas, Maceió, 2020.
url http://www.repositorio.ufal.br/handle/riufal/7315
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Alagoas
Brasil
Programa de Pós-Graduação em Ciências Farmacêuticas
UFAL
publisher.none.fl_str_mv Universidade Federal de Alagoas
Brasil
Programa de Pós-Graduação em Ciências Farmacêuticas
UFAL
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal de Alagoas (UFAL)
instname:Universidade Federal de Alagoas (UFAL)
instacron:UFAL
instname_str Universidade Federal de Alagoas (UFAL)
instacron_str UFAL
institution UFAL
reponame_str Repositório Institucional da Universidade Federal de Alagoas (UFAL)
collection Repositório Institucional da Universidade Federal de Alagoas (UFAL)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal de Alagoas (UFAL) - Universidade Federal de Alagoas (UFAL)
repository.mail.fl_str_mv ri@sibi.ufal.br
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