RESUMO
The objective of the study was to carry out phytochemical prospection through colorimetric tests to determine the groups of secondary metabolites and also to determine the total content of phenolic compounds (TPC) present in plant extracts methanol (ME), ethyl acetate (EAE), hexane (HE) and dichloromethane (DE) from the leaves of Myrsine umbellata, as well as to investigate the antimicrobial activity against twelve standard ATCC strains by the broth microdilution technique; the antioxidant potential by the DPPH method and the ABTS method and the antibiofilm potential on the biofilm biomass of standard bacteria by the crystal violet technique and tetrazolium salt reduction (MTT) assay. Phytochemical prospection detected the presence of saponins, steroids, alkaloids, anthocyanins, anthocyanidins, flavonoids, and tannins. The results of the quantitative phytochemical estimation revealed a higher content of total phenolics in DE (280.24 ± 0.037 µM GAE g ext. -1) followed by ME (159.01 ± 0.031 µM GAE g ext. -1). The ME showed the best biological activities when compared to the other extracts tested. We observed antimicrobial activity against Gram-positive Staphylococcus epidermidis strain (MIC 3.12 and MBC 6.25), antioxidant percentage of 92.58% against the DPPH radical and 420.31 µM Trolox g ext. -1 against the ABTS radical, finally showed antibiofilm action against Gram-positive strain Staphylococcus aureus, with eradication of the biomass in 92.58%. The results suggest that EM from M. umbellata represents an alternative source of plant bioactives for the development of natura products.
Assuntos
Anti-Infecciosos , Benzotiazóis , Myrsine , Ácidos Sulfônicos , Antocianinas , Antioxidantes/farmacologia , Metanol , Fenóis/farmacologia , Compostos Fitoquímicos/farmacologia , Anti-Infecciosos/farmacologiaRESUMO
The species Myrsine umbellata is a native plant of Brazil, whose barks are traditionally used in herbal medicine to treat liver disorders and combat leprosy. Therefore, the aim of the study was to identify the phytochemical prospection of ethanolic (EE) and acetonic (EA) extracts by colorimetric tests and by gas chromatography coupled to mass spectrometry (GC-MS) of the essential oil (EO) of M. umbellata leaves; evaluate the antimicrobial activity in front of standard ATCC strains by the broth microdilution technique; the antioxidant potential by DPPH reduction method and antibiofilm action by crystal violet assay and cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) based on optical density. Phytochemical prospection of EE and EA detected the presence of free steroids, alkaloids, flavonoids (flavones, flavononoids, flavonols and xanthons) and tannins in both extracts (EE and EA) and saponins only in EE. In EO, the majority compounds identified were elixene, caryophyllene (E), spatulenol, d-Cadinene and aromadendrene. EA showed antimicrobial activity with MIC and MBC/MFC values ranging from 3.12 to 100 mg.mL-1, highlighting its efficiency on the Gram-positive strain S. epidermidis. EE showed antimicrobial potential in the range of 3.12 to 200 mg.mL-1, and the Gram-negative E. coli strain was the most susceptible. However, OE showed bacteriostatic potential against S. Typhimurium, S. Abaetetuba, P. aeruginosa, and S. epidermidis strains. The ability to sequester free radicals was evident in EA extract with antioxidant activity of 89.55% and in EE with 63.05%. The antibiofilm potential was observed in EE extract which eradicated the mature biofilm biomass of all tested bacteria with high activity (50% to 84.28%) and EO also showed antibiofilm effect on mature biofilm of UEL enteroaggregative E. coli, S. aureus and S. Enteritidis strains with biomass reduction percentage of 63.74%, 68.04% and 86.19%, respectively. These results indicate the potential of M. umbellata extracts and as a source of plant bioactivity for the development of new alternative strategies for the control of planktonic or biofilm-resistant microorganisms.
Assuntos
Anti-Infecciosos , Myrsine , Óleos Voláteis , Primulaceae , Óleos Voláteis/farmacologia , Antioxidantes/farmacologia , Staphylococcus aureus , Escherichia coli , Compostos Fitoquímicos , Antibacterianos/farmacologia , Biofilmes , Extratos Vegetais/farmacologiaRESUMO
The species Myrsine umbellata is a native plant of Brazil, whose barks are traditionally used in herbal medicine to treat liver disorders and combat leprosy. Therefore, the aim of the study was to identify the phytochemical prospection of ethanolic (EE) and acetonic (EA) extracts by colorimetric tests and by gas chromatography coupled to mass spectrometry (GC-MS) of the essential oil (EO) of M. umbellata leaves; evaluate the antimicrobial activity in front of standard ATCC strains by the broth microdilution technique; the antioxidant potential by DPPH reduction method and antibiofilm action by crystal violet assay and cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) based on optical density. Phytochemical prospection of EE and EA detected the presence of free steroids, alkaloids, flavonoids (flavones, flavononoids, flavonols and xanthons) and tannins in both extracts (EE and EA) and saponins only in EE. In EO, the majority compounds identified were elixene, caryophyllene (E), spatulenol, d-Cadinene and aromadendrene. EA showed antimicrobial activity with MIC and MBC/MFC values ranging from 3.12 to 100 mg.mL-1, highlighting its efficiency on the Gram-positive strain S. epidermidis. EE showed antimicrobial potential in the range of 3.12 to 200 mg.mL-1, and the Gram-negative E. coli strain was the most susceptible. However, OE showed bacteriostatic potential against S. Typhimurium, S. Abaetetuba, P. aeruginosa, and S. epidermidis strains. The ability to sequester free radicals was evident in EA extract with antioxidant activity of 89.55% and in EE with 63.05%. The antibiofilm potential was observed in EE extract which eradicated the mature biofilm biomass of all tested bacteria with high activity (50% to 84.28%) and EO also showed antibiofilm effect on mature biofilm of UEL enteroaggregative E. coli, S. aureus and S. Enteritidis strains with biomass reduction percentage of 63.74%, 68.04% and 86.19%, respectively. These results indicate the potential of M. umbellata extracts and as a source of plant bioactivity for the development of new alternative strategies for the control of planktonic or biofilm-resistant microorganisms.
A espécie Myrsine umbellata é uma planta nativa do Brasil, tradicionalmente suas cascas são empregadas em fitoterapia no tratamento de afecções do fígado e combate a hanseníase. Portanto, o objetivo do estudo foi realizar a prospecção fitoquímica dos extratos etanólico (EE) e acetônico (EA) por meio de testes colorimétricos e por cromatografia gasosa acoplada à espectrometria de massas (CG-EM) do óleo essencial (OE) das folhas de M. umbellata; avaliar a atividade antimicrobiana frente a dozes cepas padrões ATCCs pela técnica de microdiluição em caldo; o potencial antioxidante pelo método de redução de DPPH e ação antibiofilme pela técnica do cristal violeta e a viabilidade celular foi determinada usando 3- (4,5-dimetiltiazol-2-il)-2,5-difeniltetrazólio (MTT) com base na densidade óptica. A prospecção fitoquímica dos EE e EA detectou a presença de esteroides livres, alcaloides, flavonoides (flavonas, flavononóis, flavonóis e xantonas) e taninos em ambos os extratos (EE e EA) e de saponinas somente no EE. No OE os compostos majoritários identificados foram elixeno, cariofileno (E), espatulenol, d-cadineno e aromadendreno. O EA apresentou atividade antimicrobiana com valores de CIM e CBM/CFM que variaram de 3,12 a 100 mg.mL-1, destacando sua eficiência sobre a cepa Gram-positiva S. epidermidis. Já EE apresentou potencial antimicrobiano na faixa que variou de 3,12 a 200 mg.mL-1, e a cepa Gram-negativa E. coli foi a mais suscetível. Entretanto, OE apresentou potencial bacteriostático frente às cepas S. Typhimurium, S. Abaetetuba, P. aeruginosa e S. epidermidis. A capacidade de sequestrar radicais livres foi evidenciada no extrato EA com atividade antioxidante de 89,55% e no EE com 63,05%. O potencial antibiofime foi observado no extrato EE que erradicou a biomassa do biofilme maduro de todas as bactérias testadas com elevada atividade (50% a 84,28%) e OE também apresentou efeito antibiofilme sobre o biofilme maduro das cepas E. coli enteroagreativa UEL, S. aureus e S. Enteritidis com percentual de redução de biomassa de 63,74%, 68,04% e 86,19%, respectivamente. Esses resultados indicam o potencial dos extratos e do OE de M. umbellata como fonte de bioativos vegetais para o desenvolvimento de novas estratégias alternativas para o controle de microrganismos resistentes de forma planctônica ou de biofilmes.