Ação antimicrobiana da associação dos extratos de punica granatum e curcuma longa contra cepas multirresistentes de acinetobacter baumannii
Carregando...
Data
2024-04-08
Autores
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Acinetobacter baumannii faz parte dos patógenos que representam um grande risco à saúde devido ao surgimento de cepas resistentes a agentes antimicrobianos. Portanto, a busca por estratégias alternativas para desenvolver novos compostos antibacterianos é imprescindível. Neste cenário a fitoterapia tem ganhado notoriedade, pois diversas plantas produzem naturalmente uma grande variedade de moléculas bioativas que podem ser utilizadas como agentes antimicrobianos. Diante disso, o objetivo do trabalho é avaliar a ação antimicrobiana e antibiofilme dos extratos de açafrão (Curcuma longa) e romã (Punica granatum L.) associados sobre três cepas clínicas multirresistentes de Acinetobacter baumannii. A atividade antimicrobiana sobre culturas planctônicas foi verificada por meio de microdiluição em caldo de acordo com a CLSI (norma M7-A9). Para isso, diluições seriadas dos extratos foram feitas em placas de 96 poços, em que foi adicionada a suspensão de microrganismos padronizada em 106 células/mL. Após 24h de incubação, foi verificada a turbidez dos poços, avaliando a presença da CIM (Concentração Inibitória Mínima). A partir da CIM foram semeadas alíquotas dos poços para que fosse determinada a CMM (Concentração Microbicida Mínima). A análise da associação dos extratos foi realizada por meio da técnica de microdiluição checkerboard. Os testes em biofilmes foram realizados com a distribuição de suspensões padronizadas em 107 células/mL em placas de 96 poços com caldo BHI. Após formação do biofilme, foi realizada a aplicação do extrato nas concentrações pré-determinadas pela CMM. Todos os tratamentos foram aplicados pelo tempo de 5 min e de 24h. A mensuração da viabilidade dos biofilmes após os tratamentos foi realizada pelo teste de MTT. A análise estatística foi realizada com 5% de significância (p<0,05), analisados pelo método ANOVA complementado pelo Teste de Tukey. Os extratos de C. longa e P. granatum apresentaram, respectivamente, o valor de CIM e CMM de 50 mg/mL e 0,3 mg/mL contra as cepas de Acinetobacter baumannii utilizadas, sendo assim, considerados bactericidas. Quando associados não apresentaram atividade sinérgica. Em relação a ação antibiofilme, após 5 min de contato com C. longa e P. granatum, a redução do biofilme chegou a 38% e 31% frente à cepa Ab H557. Já após 24 h, as reduções chegaram a 85% quando tratados com C. longa. Conclui-se, portanto, que extratos glicólicos isolados de C. longa P. granatum L. apresentaram efeito antimicrobiano bactericida sobre as cepas multirresistentes de Acinetobacter baumannii estudadas, tendo o extrato de C. longa apresentado melhores resultados antibiofilme.
Acinetobacter baumannii is one of the pathogens that presents a great risk to health due to the emergence of resistant strains to antimicrobial agents. Therefore, the search for alternative strategies to develop new antibacterial compounds is essential. Herbal medicine has gained notoriety, as several plants naturally produce a wide variety of bioactive molecules that can be used as antimicrobial agents. Therefore, the objective of this work is to evaluate the antimicrobial and antibiofilm action of turmeric (Curcuma longa) and pomegranate (Punica granatum L.) extracts associated on three multidrug-resistant clinical strains of Acinetobacter baumannii. Antimicrobial activity on planktonic cultures was verified by broth microdilution according to CLSI (M7-A9 standard). For this, serial dilutions of the extracts were made in 96-well plates, in which the suspension of microorganisms standardized at 106 cells/mL was added. After 24 hours of incubation, the turbidity of the wells was verified, evaluating the presence of MIC (Minimum Inhibitory Concentration). From the MIC, aliquots of the wells were seeded to determine the MMC (Minimum Microbicidal Concentration). The analysis of the association of extracts was performed using the checkerboard microdilution technique. Biofilm tests were performed with the distribution of standardized suspensions at 107 cells/mL in 96-well plates with BHI broth. After biofilm formation, the extract was applied at concentrations predetermined by the CMM. All treatments were applied for 5 min and 24 h. Measurement of biofilm viability after treatments was performed using the MTT test. Statistical analysis was performed with 5% significance (p<0.05), analyzed by the ANOVA method complemented by the Tukey test. The extracts of C. longa and P. granatum showed, respectively, the value of MIC and MMC of 50 mg/mL and 0.3 mg/mL against the Acinetobacter baumannii strains used, thus being considered bactericidal. When associated, they did not show synergistic activity. Regarding the antibiofilm action, after 5 min of contact with C. longa and P. granatum, the biofilm reduction reached 38% and 31% against the Ab H557 strain. After 24 h, the reductions reached 85% when treated with C. longa. It is concluded, therefore, that isolated glycolic extracts of C. longa and P. granatum had a bactericidal antimicrobial effect on the multidrug-resistant strains of Acinetobacter baumannii studied, with the extract of C. longa showing better antibiofilm results.
Acinetobacter baumannii is one of the pathogens that presents a great risk to health due to the emergence of resistant strains to antimicrobial agents. Therefore, the search for alternative strategies to develop new antibacterial compounds is essential. Herbal medicine has gained notoriety, as several plants naturally produce a wide variety of bioactive molecules that can be used as antimicrobial agents. Therefore, the objective of this work is to evaluate the antimicrobial and antibiofilm action of turmeric (Curcuma longa) and pomegranate (Punica granatum L.) extracts associated on three multidrug-resistant clinical strains of Acinetobacter baumannii. Antimicrobial activity on planktonic cultures was verified by broth microdilution according to CLSI (M7-A9 standard). For this, serial dilutions of the extracts were made in 96-well plates, in which the suspension of microorganisms standardized at 106 cells/mL was added. After 24 hours of incubation, the turbidity of the wells was verified, evaluating the presence of MIC (Minimum Inhibitory Concentration). From the MIC, aliquots of the wells were seeded to determine the MMC (Minimum Microbicidal Concentration). The analysis of the association of extracts was performed using the checkerboard microdilution technique. Biofilm tests were performed with the distribution of standardized suspensions at 107 cells/mL in 96-well plates with BHI broth. After biofilm formation, the extract was applied at concentrations predetermined by the CMM. All treatments were applied for 5 min and 24 h. Measurement of biofilm viability after treatments was performed using the MTT test. Statistical analysis was performed with 5% significance (p<0.05), analyzed by the ANOVA method complemented by the Tukey test. The extracts of C. longa and P. granatum showed, respectively, the value of MIC and MMC of 50 mg/mL and 0.3 mg/mL against the Acinetobacter baumannii strains used, thus being considered bactericidal. When associated, they did not show synergistic activity. Regarding the antibiofilm action, after 5 min of contact with C. longa and P. granatum, the biofilm reduction reached 38% and 31% against the Ab H557 strain. After 24 h, the reductions reached 85% when treated with C. longa. It is concluded, therefore, that isolated glycolic extracts of C. longa and P. granatum had a bactericidal antimicrobial effect on the multidrug-resistant strains of Acinetobacter baumannii studied, with the extract of C. longa showing better antibiofilm results.
Descrição
Palavras-chave
Punica granatum L., Curcuma longa, Acinetobacter baumannii, Punica granatum L., Curcuma longa, Acinetobacter baumannii