RESUMO
AIMS: We investigated the putative fungistatic and fungicidal activities of pomegranate sarcotesta lectin (PgTeL) against Cryptococcus neoformans B3501 (serotype D), specifically the ability of PgTeL to inhibit yeast capsule and biofilm formation in this strain. METHODS AND RESULTS: PgTeL showed a minimum inhibitory concentration of 172.0 µg ml-1, at which it did not exhibit a fungicidal effect. PgTeL concentrations of 4.0-256.0 µg ml-1 reduced biofilm biomass by 31.0%-64.0%. Furthermore, 32.0-256.0 µg ml-1 PgTeL decreased the metabolic activity of the biofilm by 32.0%-93.0%. Scanning electron microscopy images clearly revealed disruption of the biofilm matrix. Moreover, PgTeL disrupted preformed biofilms. At concentrations of 8.0-256.0 µg ml-1, PgTeL reduced metabolic activity in C. neoformans by 36.0%-92.0%. However, PgTeL did not inhibit the ability of B3501 cells to form capsules under stress conditions. CONCLUSIONS: PgTeL inhibited biofilm formation and disrupted preformed biofilms, demonstrating its potential for use as an anticryptococcal agent.
Assuntos
Criptococose , Cryptococcus neoformans , Punica granatum , Lectinas/farmacologia , Punica granatum/metabolismo , Plâncton/metabolismo , Biofilmes , Testes de Sensibilidade Microbiana , Antifúngicos/farmacologia , Antifúngicos/metabolismoRESUMO
The chemical composition, the antioxidant and antimicrobial potential of crude extract from leaves Cinnamomum verum and their enriched fractions was studied. Phytochemical analyses were performed by TLC and HPLC, and the antioxidant capacity was verified by DPPH⢠and ABTSâ¢+. The Minimal Inhibitory/Bactericidal Concentration was conducted against twenty-two bacteria to select five strains susceptible to extracts/fractions and resistant to the antibiotics tested. Interference of Ethyl Acetate Fraction (EAF) in resistance to synthetic antibiotic was assayed by modulatory and checkerboard model. The chromatographic data showed phenolic compounds in crude extract, as well the flavonoid enrichment in the EAF. The combination of EAF and synthetic antibiotics (ampicillin, azithromycin, ciprofloxacin, or gentamicin) provides a synergistic effect against multidrug resistant strains). The results are useful to obtain multi-targeting in a single therapy solution, which on antioxidants molecules plant-derivatives can act synergistically in antimicrobial combinations, a valuable aid as bacterial resistance modifying compounds.
Assuntos
Anti-Infecciosos , Antioxidantes , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Antioxidantes/química , Bactérias , Cinnamomum zeylanicum , Testes de Sensibilidade Microbiana , Extratos Vegetais/química , Folhas de Planta/químicaRESUMO
The aim of this study was to evaluate the phytochemical composition, antioxidant, and antimicrobial potential of crude extract and fractions of Punica granatum leaves. The extract was produced by turbo extraction, after which hexanic, ethyl acetate, and aqueous fractions were obtained by partitioning. The chemical analyses were performed by thin layer chromatography and high-performance liquid chromatography, and the antioxidant activities were assayed by DPPH. and ABTS.+ . Minimal inhibitory and bactericidal concentrations (MIC/MBC) were applied to twenty-two bacteria. Most strains susceptible to extract/fractions and resistant to antibiotics were selected, and ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with the ethyl acetate fraction (EAF) against multidrug-resistant strains in modulatory and checkboard models. The data from chromatographic analyses showed flavonoids and tannins in the extract, as well as the enrichment of EAF in phenols, mainly flavonoids. The flavonoids were connected to the electron transfer activity demonstrated in the DPPH. and ABTS.+ assays. Gram-positive strains are more susceptible to EAF. The subinhibitory concentrations of P. granatum enhanced the antimicrobial activity of the agents and reduced the EAF individual MIC, and the combination of EAF and antibiotics demonstrated a synergistic effect. These results present a promising approach for developing a therapy in which antioxidant extracts and fractions can be used in combination with antibiotics.
Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Lythraceae/química , Extratos Vegetais/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Benzotiazóis/antagonistas & inibidores , Compostos de Bifenilo/antagonistas & inibidores , Escherichia coli/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Picratos/antagonistas & inibidores , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Folhas de Planta/química , Pseudomonas aeruginosa/efeitos dos fármacos , Ácidos Sulfônicos/antagonistas & inibidoresRESUMO
The objective of this study was to conduct phytochemical characterization and biological evaluation of Hymenaea eriogyne. Crude extracts and fractions from the bark, leaves and pods, were obtained for phytochemical screening by TLC and HPLC, and evaluation of antibacterial and antioxidant potential. Chromatographic data revealed the presence of several metabolites, notably from the flavonoid class. HPLC analysis confirmed the presence of the flavanonol astilbin (taxifolin 3-O-ramnoside) and other flavonoids derived from aglycone taxifolin. In addition, it was possible to quantify phytochemical markers in the extracts and fractions, which showed an increased content of flavonoid and catechin derivatives in the fraction. Better results of the minimum inhibitory/bactericidal concentrations were obtained with extracts and fractions from bark. In the antioxidant activity using the DPPH method, the enriched bark fraction presented an IC50 of 34.46 µg/mL. These results contribute to the continuity of studies on the chemical and biological composition of the species.
Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Hymenaea , Antibacterianos/isolamento & purificação , Antioxidantes/isolamento & purificação , Hymenaea/química , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia , Casca de Planta/química , Extratos Vegetais/farmacologia , Folhas de Planta/químicaRESUMO
The sarcotesta of Punica granatum fruit contains an antimicrobial lectin called PgTeL. In this work, we evaluated the antibacterial activity of PgTeL against five drug-resistant Escherichia coli isolates able to produce ß-lactamases. Minimum inhibitory (MIC) and bactericidal (MBC) concentrations were determined by broth dilution. Morphometric and viability analyses were performed by flow cytometry, and ultrastructural changes were evaluated by scanning electron microscopy. Potential synergistic effects of PgTeL with antibiotics and anti-biofilm effect were also evaluated. PgTeL showed antibacterial activity against all isolates with MIC and MBC values ranging from 12.5 to 50.0⯵g/mL and from 25.0 to 100.0⯵g/mL, respectively. For most isolates, PgTeL postponed the growth start by at least ten hours. At the MIC, the lectin caused alterations in size, shape and structure of bacterial cells. The combination PgTeL-ceftazidime showed a synergistic effect for all isolates. Synergy was also detected with ampicillin (one isolate), carbenicillin (one isolate), cefotaxime (one isolate), cephalexin (four isolates) and cefuroxime (three isolates). PgTeL exhibited anti-biofilm activity against all isolates, causing ≥50% inhibition of biofilms at or above 6.25⯵g/mL. The antibacterial effect of PgTeL and its synergy with antibiotics indicate that this fruit-derived molecule may have potential for future treatment of multidrug-resistant infections.
Assuntos
Antibacterianos/farmacologia , Escherichia coli/efeitos dos fármacos , Lythraceae/química , Lectinas de Plantas/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Biofilmes/efeitos dos fármacos , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Escherichia coli/metabolismo , Escherichia coli/ultraestrutura , Frutas/química , Testes de Sensibilidade Microbiana , Lectinas de Plantas/química , Lectinas de Plantas/isolamento & purificação , Resistência beta-Lactâmica , beta-Lactamases/biossínteseRESUMO
The Alpinia purpurata inflorescence contains a lectin (ApuL), which has immunomodulatory activities on human cells. In the present work, it was evaluated the antibacterial and antifungal effects of ApuL against human pathogens. ApuL showed bacteriostatic activity against non-resistant (UFPEDA-02) and an oxacillin-resistant isolate (UFPEDA-672) of Staphylococcus aureus with minimal inhibitory concentrations (MIC50) of 50 and 400⯵g/mL, respectively. In addition, it showed bactericidal effect on the non-resistant isolate (minimal bactericidal concentration: 200⯵g/mL). For Candida albicans and Candida parapsilosis, ApuL showed fungistatic effect (MIC50: 200 and 400⯵g/mL, respectively). The lectin was able to impair the viability of the microorganism cells, as indicated by propidium iodide (PI) staining. Analysis of growth curves, protein leakage, and ultrastructural changes supported that ApuL acts through distinct mechanisms on S. aureus isolates. Ultrastructural analysis of ApuL-treated Candida cells revealed malformations with elongations and bulges. ApuL-oxacillin combination showed synergistic effect on the oxacillin-resistant isolates UFPEDA-670 and 671, which were not sensitive to lectin alone. Synergism was also detected for ApuL-ceftazidime against a multidrug-resistant isolate of Pseudomonas aeruginosa. Synergistic action of ApuL-fluconazole was detected for C. parapsilosis, which was insensitive to the drug alone. Biofilm formation by S. aureus non-resistant isolate and C. albicans was remarkably inhibited by ApuL at sub-inhibitory concentrations. In conclusion, ApuL showed differential effects on non-resistant and resistant bacterial isolates, was active against Candida species, and showed synergistic action in combination with antibiotics.
Assuntos
Alpinia/química , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Lectinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Candida albicans/crescimento & desenvolvimento , Candida albicans/fisiologia , Sinergismo Farmacológico , Lectinas/isolamento & purificação , Testes de Sensibilidade Microbiana , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/fisiologiaRESUMO
The pomegranate (Punica granatum) sarcotesta contains a chitin-binding lectin (PgTeL) with antibacterial activity against human pathogenic species. In this work, the structural stability of PgTeL was evaluated by fluorimetric analysis and the lectin was evaluated for cytotoxicity to human peripheral blood mononuclear cells (PBMCs) and antifungal activity against Candida albicans and Candida krusei. PgTeL folding was impaired when lectin was incubated at pH≥6.0. On the other hand, the lectin did not undergo unfolding even when heated at 100°C. PgTeL (1, 10, and 100µg/mL) was not cytotoxic to PBMCs. Antifungal activity was detected for C. albicans (MIC: 25µg/mL; MFC: 50µg/mL) and C. krusei (MIC and MFC of 12.5µg/mL). Treatment of yeast cells with PgTeL resulted in decrease of intracellular ATP content even at sub-inhibitory concentrations (½MIC and »MIC) and induced lipid peroxidation. In addition, PgTeL damaged the integrity of fungal cell wall of both species, with more pronounced effects in C. krusei. The lectin showed significant antibiofilm activity on C. albicans at sub-inhibitory concentrations (0.195 and 0.39µg/mL). In conclusion, PgTeL is an anti-Candida agent whose action mechanism involves oxidative stress, energetic collapse, damage to the cell wall and rupture of yeast cells.
Assuntos
Candida albicans/efeitos dos fármacos , Candida/efeitos dos fármacos , Lectinas/farmacologia , Lythraceae/química , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida/metabolismo , Candida/ultraestrutura , Candida albicans/metabolismo , Candida albicans/ultraestrutura , Parede Celular/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio , Lectinas/química , Lectinas/isolamento & purificação , Peroxidação de Lipídeos/efeitos dos fármacos , Testes de Sensibilidade Microbiana , TemperaturaRESUMO
This work describes the isolation of a lectin (CasuL) from the leaf pinnulae of Calliandra surinamensis and the evaluation of its cytotoxic, antimicrobial and antibiofilm properties. Proteins from pinnulae extract were precipitated with ammonium sulphate (60% saturation) and submitted to Sephadex G-75 chromatography, which yielded isolated CasuL (purification factor: 113). Native CasuL is an acidic protein (pI 5.82) with a relative molecular mass of 48kDa. This lectin is also an oligomeric protein composed of three subunits and mass spectrometry revealed similarities with a Sorghum bicolor protein. CasuL did not undergo unfolding when heated but changes in conformation and hemagglutinating activity were detected at basic pH. CasuL did not reduce the viability of human peripheral blood mononuclear cells but was toxic to leukemic K562 cells (IC50 67.04±5.78µg/mL) and breast cancer T47D cells (IC50: 58.75±2.5µg/mL). CasuL (6.25-800µg/mL) only showed bacteriostatic effect but was able to reduce biofilm formation by Staphylococcus saprophyticcus and Staphylococcus aureus (non-resistant and oxacillin-resistant isolates). CasuL showed antifungal activity against Candida krusei causing alterations in cell morphology and damage to cell wall. In conclusion, the pinnulae of C. surinamensis leaves contain a thermo-stable lectin with biotechnological potential as cytotoxic, antibiofilm, and antifungal agent.
Assuntos
Anti-Infecciosos/farmacologia , Antineoplásicos/farmacologia , Biofilmes/efeitos dos fármacos , Fabaceae/química , Folhas de Planta/química , Lectinas de Plantas/farmacologia , Anti-Infecciosos/química , Anti-Infecciosos/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Candida/efeitos dos fármacos , Candida/fisiologia , Linhagem Celular Tumoral , Humanos , Testes de Sensibilidade Microbiana , Lectinas de Plantas/química , Lectinas de Plantas/isolamento & purificação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologiaRESUMO
Aluminum salts are used as coagulants in water treatment; however, the exposure to residual aluminum has been associated with human brain lesions. The water-soluble Moringa oleifera lectin (WSMoL), which is extracted with distilled water and isolated by chitin chromatography, has coagulant activity and is able to reduce the concentration of metal ions in aqueous solutions. This study evaluated the potential of using aluminum sulfate and WSMoL to reduce the turbidity and toxicity of water from the Cavouco stream located in Recife, Pernambuco, Brazil. The water sample used (called P1) was collected from the stream source, which was found to be strongly polluted based on physicochemical and water quality analyses, as well as ecotoxicity assays with Artemia salina and seeds of Eruca sativa and Lactuca sativa. The assays combining WSMoL and aluminum sulfate were more efficient than those that used these agents separately. Furthermore, the greatest reduction in turbidity (96.8%) was obtained with the treatment using aluminum sulfate followed by WSMoL, compared to when they were applied simultaneously (91.3%). In addition, aluminum sulfate followed by WSMoL treatment resulted in residual aluminum concentration (0.3 mg/L) that was much lower than that recorded after the treatment using only the salt (35.5 mg/L). The ecotoxicity of P1 was also strongly reduced after the treatments. In summary, the combined use of aluminum sulfate and WSMoL was efficient in promoting a strong reduction of turbidity and ecotoxicity of a polluted water sample, without resulting in a high residual aluminum concentration at the conclusion of the treatment.
Assuntos
Compostos de Alúmen/química , Recuperação e Remediação Ambiental/métodos , Moringa oleifera/metabolismo , Extratos Vegetais/química , Lectinas de Plantas/química , Poluição da Água/análise , Animais , Artemia/efeitos dos fármacos , Brassicaceae/efeitos dos fármacos , Brasil , Lactuca/efeitos dos fármacos , Rios/química , Sementes/química , Água/análise , Purificação da Água/métodosRESUMO
Moringa oleifera flower extract, with trypsin inhibitor activity, is a larvicidal agent on Aedes aegypti. This work reports the isolation of trypsin inhibitor (M. oleifera flower trypsin inhibitor (MoFTI)) and its effect on A. aegypti egg hatching, viability of newly hatched larvae, survival of pupae, and growth of inhabitant bacteria from midgut of fourth-instar larvae (L4). MoFTI (K i, 2.4 µM), isolated by affinity chromatography on trypsin-agarose column, was an 18.2 kDa polypeptide on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Flower extract (at concentrations of 8.5-17.0 mg/mL) reduced egg hatchability while MoFTI (0.05-0.5 mg/mL) did not affect the hatching rate. Mortality of newly hatched larvae ranged from 3.5 to 19.1 % in the presence of the extract (4.0-17.0 mg/mL) and was also promoted by MoFTI (LC50, 0.3 mg/mL). After 72 h, larvae incubated with extract at 13.0 and 17.0 mg/mL were at stages L2 and L1, respectively, while in control they reached L3 instar. In the presence of MoFTI, at all concentrations tested, the larvae did not pass the first instar. Flower extract and MoFTI did not interfere on pupae survival. The extract and MoFTI inhibited the growth of L4 gut bacteria (minimum inhibitory concentrations of 3.47 and 0.031 mg/mL, respectively) but only the inhibitor showed bactericide effect (minimum bactericidal concentration of 1.0 mg/mL). The findings reported herein indicate that MoFTI constitutes a larvicidal principle from M. oleifera flowers against A. aegypti newly hatched larvae and is an antibacterial agent active against the microbiota from L4 gut.