AVALIAÇÃO DA ATIVIDADE ANTIFÚNGICA DO EXTRATO AQUOSO DAS FOLHAS DE Libidibia ferrea (MART. EX TUL.) L. P. QUEIROZ (PAU FERRO)

Os fungos são micro-organismos comuns no ambiente, sendo que alguns vivem de forma comensal, habitando a superfície da pele, mucosas, trato respiratório, gastrointestinal e urogenital de humanos e animais, como é o caso das espécies pertencentes ao gênero Candida. O fluconazol é um dos fármacos mais usados para combater as enfermidades causadas por esses micro-organismos, atuando em um alvo específico dos fungos, entretanto, muitas linhagens já lhe são resistentes. A resistência a medicamentos tem despertado a necessidade de procurar novas alternativas de cunho natural. Libidibia ferrea popularmente conhecida como jucá ou pau ferro é uma espécie vegetal da família Fabaceae, que possui em sua composição substâncias avaliadas quanto a seu efeito antimicrobiano. Este estudo tem como objetivo verificar o efeito antifúngico do extrato, sozinho e combinado com o Fluconazol na inibição de crescimento de Candida albicans, Candida tropicalis e Candida kruzei, bem como na virulência fúngica. Os ensaios para determinação da Concentração Inibitória de 50% dos micro-organismos (CI50), foram realizados por microdiluição (8.192 a 8 μg/mL) e na combinação extrato/fármaco, o extrato foi avaliado em concentração subinibitória (CFM/16). A Concentração Fungicida Mínima (CFM) foi verificada por subcultivo em meio sólido. O efeito inibidor sobre a morfologia fúngica foi verificado em câmaras úmidas. O extrato apresentou efeito inibidor sobre os micro-organismos em concentrações elevadas e sem relevância clínica, demonstrando um comportamento de resistência ao extrato em questão. A CI50 evidencia a potencialização do fármaco pelo extrato, onde a combinação dos dois reduziu a concentração de efeito do fluconazol frente a todas as linhagens. A CFM foi ≥ 16.384 µg/mL. Em relação a morfologia fúngica, o extrato inibiu a transição morfológica de C. albicans e C. tropicalis na concentração mais elevada. A espécie tem constituintes ativos para potencialização de efeito de fármaco comercial e para interferir na expressão de um importante fator de virulência fúngica

UPLC-MS-ESI-QTOF Analysis and Antifungal Activity of the Spondias tuberosa Arruda Leaf and Root Hydroalcoholic Extracts

The aim of this study was to identify and evaluate the chemical compositions and effects of the S. tuberosa leaf and root hydroalcoholic extracts (HELST and HERST) against different strains of Candida. Chemical analysis was performed by Ultra-Performance Liquid Chromatography Coupled to Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF). The Inhibitory Concentration of 50% of the growth (IC50) as well as the intrinsic and combined action of the extracts with the antifungal fluconazole (FCZ) were determined by the microdilution method while the minimum fungicidal concentrations (MFCs) and the effect on fungal morphological transitions were analyzed by subculture and in humid chambers, respectively. From the preliminary phytochemical analysis, the phenols and flavonoids were the most abundant. The intrinsic IC50 values for HELST ranged from 5716.3 to 7805.8 µg/mL and from 6175.4 to 51070.9 µg/mL for the HERST, whereas the combination of the extracts with fluconazole presented IC50 values from 2.65 to 278.41 µg/mL. The MFC of the extracts, individually, for all the tested strains was ≥16384 µg/mL. When fluconazole was combined with each extract, the MFC against CA URM 5974 was reduced (HELST: 2048 and HERST: 4096 µg/mL). Synergism was observed against standard C. albicans (CA) and C. tropicalis (CT) strains and with the root extract against the CT isolate. The leaf extract inhibited the morphological transition of all strains while the root extract inhibited only CT strains

Análise química e efeito antifúngico de Spondias tuberosa Arruda (umbu) frente a leveduras do gênero Candida

Infecções causadas por fungos tem se tornado um problema cada vez maior de saúde pública e este fato vem se agravando por conta do surgimento de novos mecanismos de resistência. Diante disso, tem se buscado nos produtos naturais, uma alternativa no combate a esses patógenos. Spondias tuberosa Arruda (umbu) é uma planta do semiárido brasileiro, de importância na medicina tradicional. Baseado nesse contexto e na importância da conservação da espécie, esse trabalho objetivou analisar a composição química e avaliar de forma comparativa a ação antifúngica e o potencial inibidor de virulência dos extratos aquosos e hidroalcoólicos das folhas e raízes de Spondias tuberosa Arruda (umbu) frente às linhagens do gênero Candida. A análise dos componentes químicos das folhas e raízes foi feito por UPLC-ESI-QTOF-MS. Os ensaios antifúngicos foram realizados frente à C. albicans e C. tropicalis (tipo padrão e isolado clínico), por microdiluição, para determinação da IC50 e obtenção da curva de viabilidade celular sob a ação intrínseca dos extratos e do fluconazol (controle) e de forma combinada extrato/fármaco. A Concentração Fungicida Mínima (CFM) foi realizada por subcultivo. Para verificação do efeito sobre a morfologia das leveduras e inibição de sua virulência foram preparadas câmaras de microcultivo. Na identificação dos extratos constatou-se principalmente a presença de composto fenólicos e flavonoides. Os extratos apresentaram um efeito fungistático (≥ 16.384 μg/mL) e, na combinação destes com o Fluconazol, em algumas concentrações, o efeito foi sinérgico. Na transição morfológica os extratos conseguiram inibir ou reduzir os filamentos das cepas testadas em concentrações progressivas. Os resultados obtidos mostraram que os extratos das folhas tem efeito semelhantes com os das raízes, e assim pode ser uma alternativa de diminuir o impacto causado pela utilização das raízes, procurando contribuir com a conservação e manejo da espécie.Infections caused by fungi have become an increasing public health problem and this fact has been aggravated by the emergence of new resistance mechanisms. Given this, natural products have been sought as an alternative in combating these pathogens. Spondias tuberosa Arruda (Umbu) is a Brazilian semiarid plant of importance in traditional medicine. Based on this context and the importance of conservation of the species, this work aimed to analyze the chemical composition and comparatively evaluate the antifungal action and the potential virulence inhibitor of aqueous and hydroalcoholic extracts of leaves and roots of Spondias tuberosa Arruda (umbu) against strains of the genus Candida. The analysis of the chemical components of leaves and roots was performed by UPLC-ESI-QTOF-MS. Antifungal assays were performed against C. albicans and C. tropicalis (standard and isolated clinical type) by microdilution to determine the IC50 and to obtain the cell viability curve under the intrinsic action of extracts and fluconazole (control) and extract / drug combination form. Minimum Fungicidal Concentration (MFC) was performed by subculture. To verify the effect on yeast morphology and inhibition of yeast virulence, microculture chambers were prepared. In the identification of the extracts it was verified mainly the presence of phenolic compounds and flavonoids. The extracts had a fungistatic effect (≥ 16,384 μg / mL) and, in their combination with Fluconazole, at some concentrations, the effect was synergistic. In the morphological transition the extracts were able to inhibit or reduce the filaments of the tested strains in progressive concentrations. The obtained results showed that the leaf extracts have similar effect to the roots, and thus can be an alternative to reduce the impact caused by the use of the roots, seeking to contribute to the conservation and management of the species.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESConselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNP

Evaluación de la actividad antibacteriana y moduladora del extracto etanólico de Calotropis procera (Aiton) W.T. Aiton contra cepas bacterianas multirresistentes

El objetivo de este estudio fue evaluar la actividad antibacteriana y moduladora del extracto etanólico de Calotropis procera (Aiton) W.T. Aiton contra cepas multirresistentes de bacterias. Por el método de microdilución, fueron definidas la concentración inhibidora mínima (MIC) del extracto y la modulación con la concentración inhibidora CIM / 8 del extracto con norfloxacina, gentamicina e imipenem contra Staphylococcus aureus, Escherichia coli y Pseudomonas aeruginosa. Se obtuvo 512 μg/mL para g/mL para Pseudomonas aeruginosa. Se descubrió sinergismo en el caso de Staphylococcus aureus, en la modulación con norfloxacina y gentamicina, mientras que con imipenem frente a Pseudomonas aeruginosa y con gentamicina para Escherichia coli. Con base en estos resultados, se necesitan más estudios para probar la actividad antibacteriana del extracto.The objective of this study was to evaluate the antibacterial and modulatory activities of ethanolic extract of Calotropis procera (Aiton) W.T. Aiton against resistant species. By microdilution method, the minimum inhibitory concentration (MIC) of the extract and modulation of the subinhibitory concentration MIC/8 to norfloxacine, gentamicin and imipenem against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. There was obtained 512 μg/mL para g/mL to Pseudomonas aeruginosa. To Staphylococcus aureus, modulation showed synergism to norfloxacin and gentamicin, with imipenem against Pseudomonas aeruginosa and gentamicin against Escherichia coli. Based on these results, more studies are needed to test the antibacterial activity of the extract

AVALIAÇÃO DO POTENCIAL ANTIFÚNGICO E AÇÃO MODULADORA DO EXTRATO AQUOSO DAS FOLHAS DE Ricinus communis L. (Mamona) SOBRE CEPAS DE Candida spp.

Infecções fúngicas ocasionadas por espécies de Candida são consideradas graves problemas patológicos. A mamona (Ricinus communis L.) é apontada como espécie versátil detentora de várias propriedades medicinais, entre estas, tratamento de infecções de pele e diarreia. Este trabalho objetivou avaliar a atividade antifúngica do Extrato Aquoso das Folhas de R. communis L. (EAFRC). Na metodologia aplicada para os testes, empregou-se a microdiluição em microplacas para estabelecer a Concentração Inibitória de 50% dos micro-organismos (CI50) dos produtos isolados e para os testes de modulação em combinação com fluconazol, com resultados obtidos através da leitura em espectrofotômetro com leitor de ELISA. O subcultivo em placa de Petri usado para determinar a Concentração Fungicida Mínima (CFM). Os resultados não apresentaram CFM para as concentrações testadas do extrato, a CIM foi estabelecida em uma concentração bastante elevada do produto natural, nos testes com o produto natural também não houve a constatação de IC50, não havendo uma concentração capaz de inibir 50% dos micro-organismos. Nos resultados da modulação do fluconazol o EAFRC mostrou-se antagônico reduzindo o efeito do fármaco quando combinado a este. Os resultados obtidos, entretanto, não excluem a possibilidade de a planta possuir propriedades fungicidas, pois uma mesma planta pode apresentar resultados diferentes na produção de extratos a partir de suas raízes e caules, ou quando usados contra linhagens fúngicas diferentes das testadas, sendo assim os estudos sobre os potenciais da planta devem continuar para se estabelecer novos resultados

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth (Lamiaceae) against Candida strains

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration (MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction (DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥ 4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC50 of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development

Phytochemical Analysis, Antibacterial Activity and Modulating Effect of Essential Oil from Syzygium cumini (L.) Skeels

One of the main global problems that affect human health is the development of bacterial resistance to different drugs. As a result, the growing number of multidrug-resistant pathogens has contributed to an increase in resistant infections and represents a public health problem. The present work seeks to investigate the chemical composition and antibacterial activity of the essential oil of Syzygium cumini leaves. To identify its chemical composition, gas chromatography coupled to mass spectrometry was used. The antibacterial activity test was performed with the standard strains Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 25853 and Staphylococcus aureus ATCC 25923 and multidrug-resistant clinical isolates E. coli 06, P. aeruginosa 24 and S. aureus 10. The minimum inhibitory concentration (MIC) was determined by serial microdilution as well as the verification of the modulating effect of the antibiotic effect. In this test, the oil was used in a subinhibitory concentration. The test reading was performed after 24 h of incubation at 37 °C. The results show that the major chemical constituent is α-pinene (53.21%). The oil showed moderate activity against E. coli ATCC 25922, with the MIC of 512 µg/mL; there was no activity against the other strains. The oil potentiated the effect of antibiotics demonstrating possible synergism when associated with gentamicin, erythromycin and norfloxacin against E. coli 06 and S. aureus 10

Physico-chemical characterization and antibacterial activity of inclusion complexes of Hyptis martiusii Benth essential oil in β-cyclodextrin

Bezerra, Daniel Pereira. Fundação Gonçalo Moniz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil. a Pharmacy Department, Federal University of Sergipe, Sergipe, Brazil b Department of Biological Chemistry, Regional University of Cariri, Crato, Ceará, Brazil c Physiology Department, Federal University of Sergipe, Sergipe, Brazil d Gonçalo Muniz Research Center, Oswaldo Cruz Foundation, Bahia, Brazil e Department of Mineralogy and Geotectonics, Institute of Geosciences, University of SãoPaulo-SP, BrazilSubmitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-04-23T13:36:40Z No. of bitstreams: 1 Andrade TA Physico-chemical....pdf: 3585852 bytes, checksum: 8a7e1bcbe17cb62a62c347459f8804ba (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-04-23T13:48:34Z (GMT) No. of bitstreams: 1 Andrade TA Physico-chemical....pdf: 3585852 bytes, checksum: 8a7e1bcbe17cb62a62c347459f8804ba (MD5)Made available in DSpace on 2018-04-23T13:48:34Z (GMT). No. of bitstreams: 1 Andrade TA Physico-chemical....pdf: 3585852 bytes, checksum: 8a7e1bcbe17cb62a62c347459f8804ba (MD5) Previous issue date: 2017Conselho Nacional de Desenvolvimento Científico e Tecnológico/CNPq/Brazil and Fundação de Amparo à Pesquisa do Estado de Sergipe/FAPITEC-SE.Múltipla – ver em NotasCyclodextrins (CDs) have been used as important pharmaceutical excipients for improve the physicochemical properties of the drugs of low solubility as the essential oil of Hyptis martiusii. This oil is important therapeutically, but the low solubility and bioavailability compromises your use. Therein, the aim of this study was to obtain and to characterize physico-chemically the samples obtained by physical mixture (PM), paste complexation (PC) and slurry complexation (SC) of the essential oil Hyptis martiusii (EOHM) in β-CD, and to compare the antibacterial and modulatory-antibiotic activity of products obtained and oil free. The physicochemical characterization was performed by differential scanning calorimetry (DSC), thermogravimetry/derivative thermogravimetry (TG/DTG), scanning electron microscopy (SEM), X-ray diffraction (XRD) and Karl Fischer titration. Additionally, the antibacterial tests were performed by microdilution technique. Thus, it was observed that the PM method showed low complexing capacity, unlike PC and SC in which it was observed the formation of inclusion complexes. In addition, the second stage of the TG/DTG curves showed that SC was the best method inclusion with mass loss of 6.9% over the PC that was 6.0%. The XRD results corroborate with the results above suggesting the formation of new solid phase and the SEM photomicrographs showed the porous surface of the samples PC and SC. The essential oil alone demonstrated an antibacterial and modulatory effect against the S. aureus and the Gram negative strain, respectively. However, the β-CD and the inclusion complex did not demonstrate any biological activity in the performed antibacterial assays

UPLC-MS-ESI-QTOF Analysis and Antifungal Activity of Aqueous Extracts of Spondias tuberosa

This study aimed to identify the chemical composition of the Spondias tuberosa aqueous leaf and root extracts (EALST and EARST) and to evaluate their effect, comparatively, against opportunistic pathogenic fungi. Ultra-Performance Liquid Chromatography Coupled to a Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF) was employed for chemical analysis. Candida albicans and C. tropicalis standard strains and clinical isolates were used (CA INCQS 40006, CT INCQS 40042, CA URM 5974, and CT URM 4262). The 50% Inhibitory Concentration for the fungal population (IC50) was determined for both the intrinsic action of the extracts and the extract/fluconazole (FCZ) associations. The determination of the Minimum Fungicidal Concentration (MFC) and the verification of effects over fungal morphological transitions were performed by subculture in Petri dishes and humid chambers, respectively, both based on micro-dilution. UPLC-MS-ESI-QTOF analysis revealed the presence of phenolic and flavonoid compounds. The association of the extracts with fluconazole, resulted in IC50 values from 2.62 µg/mL to 308.96 µg/mL. The MFC of the extracts was ≥16,384 µg/mL for all tested strains, while fluconazole obtained an MFC of 8192 µg/mL against C. albicans strains. A reduction in MFC against CA URM 5974 (EALST: 2048 µg/mL and EARST: 1024 µg/mL) occurred in the extract/fluconazole association

Effect of α-Bisabolol and Its β-Cyclodextrin Complex as TetK and NorA Efflux Pump Inhibitors in Staphylococcus aureus Strains

Efflux pumps are proteins present in the plasma membrane of bacteria, which transport antibiotics and other compounds into the extracellular medium, conferring resistance. The discovery of natural efflux pump inhibitors is a promising alternative. α-Bisabolol is a sesquiterpene isolated from several plants such as Matricaria chamomilla L. and has important properties such as antibacterial and anti-inflammatory activity. Currently, the formation of inclusion complexes with β-Cyclodextrin has been used for improving the physicochemical characteristics of the host molecule. This study evaluated the effect of α-Bisabolol, in isolation and in complexation with β-Cyclodextrin, as TetK and NorA efflux pump inhibitors in Staphylococcus aureus strains. The minimum inhibitory concentration (MIC) was determined. Subsequently, inhibitory activity over the pumps was observed by an MIC reduction for the antibiotics, by using subinhibitory concentrations (MIC/8) in combination with tetracycline and norfloxacin. The MIC of the compounds was ≥1024 μg/mL. α-Bisabolol potentiated the action of tetracycline and reduced the MIC of norfloxacin to a clinically relevant concentration. The complexed substance showed synergism however, the effect of the isolated α-Bisabolol was superior to the complex. These results indicate α-Bisabolol is a potential substance to be used as an efflux pump inhibitor