Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação

Detalhes bibliográficos
Autor(a) principal: Carvalhais, Bárbara Ellen Santos
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/10829
Resumo: Stenotrophomas maltophilia is a non-fermenting carbohydrate Gram-negative bacillus, which although of environmental origin, has been reported in bacteremias related to catheter use, urinary tract infections, endocarditis, and respiratory tract infections, especially in immunosuppressed patients. In addition to the intrinsic resistance to several antimicrobials, another characteristic of S. maltophilia that makes it an important opportunistic pathogen is its ability to form biofilms. Thus, our objective was to evaluate the effect of antimicrobials indicated for the treatment of S. maltophilia infections on biofilm in different stages of formation (adhesion, exponential growth and maturation). We evaluated 19 clinical samples and one reference sample (ATCC 13637) of S. maltophilia identified by biochemical methods and confirmed by amplification of the 23S region of ribosomal RNA. Samples were evaluated for similarity by ERIC-PCR. After this initial characterization, the samples were submitted to the following tests: (i) biofilm formation capacity; (ii) biofilm formation kinetics; (iii) antimicrobial susceptibility profile to chloramphenicol (CHL), levofloxacin (LVX), ceftazidime (CAZ), sulfamethoxazole-trimethoprim (SXT) and gentamicin (GEN) by the broth microdilution method and (iv) determination of the minimum inhibitory concentration of antimicrobial agents against biofilm (MIC-b) at different stages of formation. The samples showed genetic heterogeneity in the technique used. All were moderate producers of biofilm, showing adhesion until the 4th hour of incubation, exponential growth between the 4th and 8th hour and mature biofilm between 8 and 24 hours of incubation. All clinical samples in their planktonic forms were sensitive to LVX and SXT and most were sensitive to CHL. Higher resistance frequencies were for CAZ and GEN. We observed that bacterial susceptibility decreases as the biofilm becomes mature. The antimicrobial with the best biofilm activity regardless of the formation time was GEN followed by LVX. SXT which is the first choice antimicrobial for the treatment of S. maltophilia infections was the least effective against biofilms along with CAZ. The determination of the susceptibility profile of the samples in the biofilm shows that GEN was the best antimicrobial activity at all formation times. CAZ was the least effective against 24 hour biofilm and SXT was the least effective in 4 and 6 hour biofilm. Considering the MIC-b/MIC ratio, we can organize antimicrobials in the following decreasing order of activity against mature (24h) biofilms: GEN, LVX, CHL, SXT and CAZ. Based on these findings, we conclude that GEN may be a good option against infections related to S. maltophilia biofilms in regions where this microorganism is susceptible to aminoglycosides. LVX remains a good option against S. maltophilia biofilms. On the other hand, SXT that is currently considered the gold standard for infection by S. maltophilia should not be used in infections where biofilms may be present.
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spelling Santos, Kênia Valéria dosCarvalhais, Bárbara Ellen SantosSchuenck, Ricardo PintoSpano, Liliana Cruz2019-03-11T12:49:32Z2019-03-112019-03-11T12:49:32Z2018-12-06Stenotrophomas maltophilia is a non-fermenting carbohydrate Gram-negative bacillus, which although of environmental origin, has been reported in bacteremias related to catheter use, urinary tract infections, endocarditis, and respiratory tract infections, especially in immunosuppressed patients. In addition to the intrinsic resistance to several antimicrobials, another characteristic of S. maltophilia that makes it an important opportunistic pathogen is its ability to form biofilms. Thus, our objective was to evaluate the effect of antimicrobials indicated for the treatment of S. maltophilia infections on biofilm in different stages of formation (adhesion, exponential growth and maturation). We evaluated 19 clinical samples and one reference sample (ATCC 13637) of S. maltophilia identified by biochemical methods and confirmed by amplification of the 23S region of ribosomal RNA. Samples were evaluated for similarity by ERIC-PCR. After this initial characterization, the samples were submitted to the following tests: (i) biofilm formation capacity; (ii) biofilm formation kinetics; (iii) antimicrobial susceptibility profile to chloramphenicol (CHL), levofloxacin (LVX), ceftazidime (CAZ), sulfamethoxazole-trimethoprim (SXT) and gentamicin (GEN) by the broth microdilution method and (iv) determination of the minimum inhibitory concentration of antimicrobial agents against biofilm (MIC-b) at different stages of formation. The samples showed genetic heterogeneity in the technique used. All were moderate producers of biofilm, showing adhesion until the 4th hour of incubation, exponential growth between the 4th and 8th hour and mature biofilm between 8 and 24 hours of incubation. All clinical samples in their planktonic forms were sensitive to LVX and SXT and most were sensitive to CHL. Higher resistance frequencies were for CAZ and GEN. We observed that bacterial susceptibility decreases as the biofilm becomes mature. The antimicrobial with the best biofilm activity regardless of the formation time was GEN followed by LVX. SXT which is the first choice antimicrobial for the treatment of S. maltophilia infections was the least effective against biofilms along with CAZ. The determination of the susceptibility profile of the samples in the biofilm shows that GEN was the best antimicrobial activity at all formation times. CAZ was the least effective against 24 hour biofilm and SXT was the least effective in 4 and 6 hour biofilm. Considering the MIC-b/MIC ratio, we can organize antimicrobials in the following decreasing order of activity against mature (24h) biofilms: GEN, LVX, CHL, SXT and CAZ. Based on these findings, we conclude that GEN may be a good option against infections related to S. maltophilia biofilms in regions where this microorganism is susceptible to aminoglycosides. LVX remains a good option against S. maltophilia biofilms. On the other hand, SXT that is currently considered the gold standard for infection by S. maltophilia should not be used in infections where biofilms may be present.Stenotrophomas maltophilia é um bacilo Gram-negativo não fermentador de carboidratos, que, embora seja de origem ambiental, tem sido relatado em bacteremias relacionadas ao uso de cateter, infecções das vias urinárias, endocardites, e infecções do trato respiratório, especialmente em pacientes imunossuprimidos. Além da resistência intrínseca a diversos antimicrobianos, outra característica de S. maltophilia que a torna um importante patógeno oportunista é sua capacidade de formação de biofilmes. Assim, nosso objetivo foi avaliar o efeito dos antimicrobianos indicados para o tratamento de infecções por S. maltophilia sobre o biofilme em diferentes fases de formação (adesão, crescimento exponencial e maturação). Foram avaliadas 19 amostras clínicas e uma amostra de referência (ATCC 13637) de S. maltophilia identificadas por métodos bioquímicos e confirmadas por amplificação da região 23S do RNA ribossômico. As amostras foram avaliadas quanto à similaridade por ERIC-PCR. Após esta caracterização inicial, as amostras foram submetidas aos seguintes testes: (i) capacidade de formação de biofilme; (ii) cinética de formação do biofilme; (iii) perfil de suscetibilidade aos antimicrobianos cloranfenicol (CHL), levofloxacina (LVX), ceftazidima (CAZ), sulfametoxazol-trimetoprim (SXT) e gentamicina (GEN) pelo método de microdiluição em caldo e (iv) determinação da concentração inibitória mínima dos antimicrobianos contra biofilme (MIC-b) em diferentes estágios de formação. As amostras mostraram heterogeneidade genética na técnica utilizada. Todas foram produtoras moderadas de biofilme, mostrando aderência até a 4ª hora de incubação, crescimento exponencial entre a 4ª e 8ª hora e biofilme maduro entre 8 e 24 horas de incubação. Todas as amostras clínicas em suas formas plactônicas foram sensíveis à LVX e ao SXT e a maioria foi sensível ao CHL. Maiores frequências de resistência foram para CAZ e GEN. Observamos que a susceptibilidade bacteriana diminui à medida que o biofilme se torna maduro. O antimicrobiano com melhor atividade em biofilme, independentemente do tempo de formação foi GEN seguido de LVX. SXT que é o antimicrobiano de 1ª escolha para o tratamento de infecções por S. maltophilia foi o menos efetivo contra biofilmes, juntamente com CAZ. A determinação do perfil de suscetibilidade das amostras no biofilme mostra que GEN foi o antimicrobiano com melhor atividade em todos os tempos de formação. CAZ foi o menos eficaz contra o biofilme de 24 horas e SXT foi o menos eficaz em biofilme de 4 e 6 horas. Considerando a razão MIC-b/MIC podemos organizar os antimicrobianos na seguinte ordem decrescente de atividade contra biofilmes de maduros (24h): GEN, LVX, CHL, SXT e CAZ. Com base nestes achados, concluímos que GEN pode ser uma boa opção contra infecções relacionadas a biofilmes de S. maltophilia em regiões onde este microrganismo apresente susceptibilidade aos aminoglicosídeos. LVX continua sendo uma boa opção contra biofilmes de S. maltophilia. Por outro lado, SXT que atualmente é considerado o padrão ouro para infecção por S. maltophilia não deve ser usado em infecções onde biofilmes possam estar presentesTexthttp://repositorio.ufes.br/handle/10/10829porUniversidade Federal do Espírito SantoMestrado em Ciências FarmacêuticasPrograma de Pós-Graduação em Ciências FarmacêuticasUFESBRCentro de Ciências da SaúdeBiofilmAntimicrobialsAgentes antiinfecciososStenotrophomonas maltophiliaBiofilmeAntimicrobianosMIC-bFarmácia615Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formaçãoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESORIGINALtese_12813_Dissertação Final - Bárbara Ellen Santos Carvalhais.pdftese_12813_Dissertação Final - Bárbara Ellen Santos Carvalhais.pdfapplication/pdf1421896http://repositorio.ufes.br/bitstreams/bfb7b574-eedc-45f3-b37d-0053b0d1b438/downloaddf97350e7adeec0d605915f2831bea0eMD5210/108292024-06-27 11:00:15.162oai:repositorio.ufes.br:10/10829http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-06-27T11:00:15Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false
dc.title.none.fl_str_mv Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
title Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
spellingShingle Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
Carvalhais, Bárbara Ellen Santos
Biofilm
Antimicrobials
Agentes antiinfecciosos
Stenotrophomonas maltophilia
Biofilme
Antimicrobianos
MIC-b
Farmácia
615
title_short Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
title_full Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
title_fullStr Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
title_full_unstemmed Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
title_sort Efeito de antimicrobianos sobre biofilmes de stenotrophomonas maltophilia em diferentes estágios de formação
author Carvalhais, Bárbara Ellen Santos
author_facet Carvalhais, Bárbara Ellen Santos
author_role author
dc.contributor.advisor1.fl_str_mv Santos, Kênia Valéria dos
dc.contributor.author.fl_str_mv Carvalhais, Bárbara Ellen Santos
dc.contributor.referee1.fl_str_mv Schuenck, Ricardo Pinto
dc.contributor.referee2.fl_str_mv Spano, Liliana Cruz
contributor_str_mv Santos, Kênia Valéria dos
Schuenck, Ricardo Pinto
Spano, Liliana Cruz
dc.subject.eng.fl_str_mv Biofilm
Antimicrobials
topic Biofilm
Antimicrobials
Agentes antiinfecciosos
Stenotrophomonas maltophilia
Biofilme
Antimicrobianos
MIC-b
Farmácia
615
dc.subject.por.fl_str_mv Agentes antiinfecciosos
Stenotrophomonas maltophilia
Biofilme
Antimicrobianos
MIC-b
dc.subject.cnpq.fl_str_mv Farmácia
dc.subject.udc.none.fl_str_mv 615
description Stenotrophomas maltophilia is a non-fermenting carbohydrate Gram-negative bacillus, which although of environmental origin, has been reported in bacteremias related to catheter use, urinary tract infections, endocarditis, and respiratory tract infections, especially in immunosuppressed patients. In addition to the intrinsic resistance to several antimicrobials, another characteristic of S. maltophilia that makes it an important opportunistic pathogen is its ability to form biofilms. Thus, our objective was to evaluate the effect of antimicrobials indicated for the treatment of S. maltophilia infections on biofilm in different stages of formation (adhesion, exponential growth and maturation). We evaluated 19 clinical samples and one reference sample (ATCC 13637) of S. maltophilia identified by biochemical methods and confirmed by amplification of the 23S region of ribosomal RNA. Samples were evaluated for similarity by ERIC-PCR. After this initial characterization, the samples were submitted to the following tests: (i) biofilm formation capacity; (ii) biofilm formation kinetics; (iii) antimicrobial susceptibility profile to chloramphenicol (CHL), levofloxacin (LVX), ceftazidime (CAZ), sulfamethoxazole-trimethoprim (SXT) and gentamicin (GEN) by the broth microdilution method and (iv) determination of the minimum inhibitory concentration of antimicrobial agents against biofilm (MIC-b) at different stages of formation. The samples showed genetic heterogeneity in the technique used. All were moderate producers of biofilm, showing adhesion until the 4th hour of incubation, exponential growth between the 4th and 8th hour and mature biofilm between 8 and 24 hours of incubation. All clinical samples in their planktonic forms were sensitive to LVX and SXT and most were sensitive to CHL. Higher resistance frequencies were for CAZ and GEN. We observed that bacterial susceptibility decreases as the biofilm becomes mature. The antimicrobial with the best biofilm activity regardless of the formation time was GEN followed by LVX. SXT which is the first choice antimicrobial for the treatment of S. maltophilia infections was the least effective against biofilms along with CAZ. The determination of the susceptibility profile of the samples in the biofilm shows that GEN was the best antimicrobial activity at all formation times. CAZ was the least effective against 24 hour biofilm and SXT was the least effective in 4 and 6 hour biofilm. Considering the MIC-b/MIC ratio, we can organize antimicrobials in the following decreasing order of activity against mature (24h) biofilms: GEN, LVX, CHL, SXT and CAZ. Based on these findings, we conclude that GEN may be a good option against infections related to S. maltophilia biofilms in regions where this microorganism is susceptible to aminoglycosides. LVX remains a good option against S. maltophilia biofilms. On the other hand, SXT that is currently considered the gold standard for infection by S. maltophilia should not be used in infections where biofilms may be present.
publishDate 2018
dc.date.issued.fl_str_mv 2018-12-06
dc.date.accessioned.fl_str_mv 2019-03-11T12:49:32Z
dc.date.available.fl_str_mv 2019-03-11
2019-03-11T12:49:32Z
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dc.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Ciências Farmacêuticas
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dc.publisher.department.fl_str_mv Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Ciências Farmacêuticas
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