Anti-Candida activity of the water-soluble lectin from Moringa oleifera seeds (WSMoL).

This work reports the effects of the water-soluble lectin from Moringa oleifera seeds (WSMoL) on growth and survival of Candida species. In addition, cellular alterations linked to the antifungal effect were investigated. The minimal inhibitory (MIC) and fungicidal (MFC) concentrations were determined and 24-h growth curves in absence and presence of lectin were established. Flow cytometry was used to evaluate the induction of apoptosis/necrosis, alterations in mitochondrial membrane potential (ΔΨm), and occurrence of lysosomal damage. WSMoL inhibited the growth of C. albicans, C. glabrata, C. krusei and C. parapsilosis with MIC of 20µg/mL. The lowest MFC (20µg/mL) was detected for C. glabrata and the highest (80µg/mL) for C. albicans and C. parapsilosis. The inhibitory effect started from the ninth to nineteenth hour of incubation depending on the fungal species. Incubation with the lectin at the MIC for 24h increased the number of cells undergoing apoptosis and necrosis. Hyperpolarization of the mitochondrial membrane was detected after 12-h treatment, followed by reduction of ΔΨm or depolarization after 24h. No lysosomal damage was detected in treated cells. In conclusion, WSMoL is a fungistatic and fungicide agent against Candida with differential effects depending on the species.

Multi-effect of the water-soluble Moringa oleifera lectin against Serratia marcescens and Bacillus sp.: antibacterial, antibiofilm and anti-adhesive properties.

AIMS: To evaluate the antibiofilm potential of water-soluble Moringa oleifera seed lectin (WSMoL) on Serratia marcescens and Bacillus sp. METHODS AND RESULTS: WSMoL inhibited biofilm formation by S. marcescens at concentrations lower than 2·6 µg ml-1 and impaired bacterial growth at higher concentrations, avoiding biofilm formation. For Bacillus sp., the lectin inhibited bacterial growth at all concentrations. The antibiofilm action of WSMoL is associated with damage to bacterial cells. WSMoL did not disrupt preformed S. marcescens biofilms but was able to damage cells inside them. On the other hand, the lectin reduced the number of cells in Bacillus sp. biofilm treated with it. WSMoL was able to control biofilm formation when immobilized on glass surface (116 µg cm-2 ), damaging S. marcescens cells and avoiding adherence of Bacillus sp. cells on glass. The Bacillus sp. isolate is member of Bacillus subtilis species complex and closely related to species of the conspecific 'amyloliquefaciens' group. CONCLUSION: WSMoL prevented biofilm development by S. marcescens and Bacillus sp. and the antibiofilm effect is also observed when the lectin is immobilized on glass. SIGNIFICANCE AND IMPACT OF THE STUDY: Taking together, our results provide support to the potential use of WSMoL for controlling biofilm formation by bacteria.

Water-soluble Moringa oleifera lectin interferes with growth, survival and cell permeability of corrosive and pathogenic bacteria.

AIMS: This work evaluated the antibacterial activity of a water-soluble Moringa oleifera seed lectin (WSMoL) by evaluating its effect on growth, survival and cell permeability of Bacillus sp., Bacillus cereus, Bacillus pumillus, Bacillus megaterium, Micrococcus sp., Pseudomonas sp., Pseudomonas fluorescens, Pseudomonas stutzeri and Serratia marcescens. In addition, the effect of lectin on membrane integrity of most sensitive species was also evaluated. All the tested bacteria are able to cause biocorrosion and some are also responsible for human infections. METHODS AND RESULTS: WSMoL inhibited the bacterial growth, induced agglutination and promoted the leakage of proteins from cells of all strains. Bactericidal effect was detected against Bacillus sp., B. pumillus, B. megaterium, Ps. fluorescens and Ser. marcescens. The bacteriostatic effect of lectin was evident with only 6 h of incubation. Fluorescence microscopy of Ser. marcescens showed that WSMoL caused loss of cell integrity and indicated an anti-biofilm activity of the lectin. CONCLUSIONS: WSMoL was active against the bacteria by inhibiting growth and affecting cell permeability. The lectin also interfered with membrane integrity of Ser. marcescens, the most sensitive species. SIGNIFICANCE AND IMPACT OF THE STUDY: The study indicates that WSMoL was active against bacteria that cause serious problems in both industrial and health sectors. Also, the study contributes for the 'state-of-art' on antibacterial mechanisms of lectins.

Antimicrobial lectin from Schinus terebinthifolius leaf.

AIMS: Schinus terebinthifolius leaves are used for treating human diseases caused by micro-organisms. This work reports the isolation, characterization and antimicrobial activity of S. terebinthifolius leaf lectin (SteLL). METHODS AND RESULTS: The isolation procedure involved protein extraction with 0.15 mol l(-1) NaCl, filtration through activated charcoal and chromatography of the filtrate on a chitin column. SteLL is a 14-kDa glycopeptide with haemagglutinating activity that is inhibited by N-acetyl-glucosamine, not affected by ions (Ca(2+) and Mg(2+)) and stable upon heating (30-100 °C) as well as over the pH 5.0-8.0. The antimicrobial effect of SteLL was evaluated by determining the minimal inhibitory (MIC), bactericide (MBC) and fungicide (MFC) concentrations. Lectin was active against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella enteritidis and Staphylococcus aureus. Highest bacteriostatic and bactericide effects were detected for Salm. enteritidis (MIC: 0.45 µg ml(-1)) and Staph. aureus (MBC: 7.18 µg ml(-1)), respectively. SteLL impaired the growth (MIC: 6.5 µg ml(-1)) and survival (MFC: 26 µg ml(-1)) of Candida albicans. CONCLUSIONS: SteLL, a chitin-binding lectin, purified in milligram quantities, showed antimicrobial activity against medically important bacteria and fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: SteLL can be considered as a new biomaterial for potential antimicrobial applications.

Biological Activities of Libidibia (Caesalpinia) ferrea var. parvifolia (Mart. ex Tul.) L. P. Queiroz Pod Preparations.

Libidibia ferrea has been used in folk medicine throughout Brazil, and this study evaluated the biological activities of crude extract (CE) as well as a partially purified fraction (F80) obtained from its pods. Results from the MTT assay revealed that only F80 inhibited NCI-H292 cell growth; however, neither CE nor F80 reduced HEp-2 cell growth or sarcoma 180 tumor weight with the in vivo assay. Acute oral toxicity of the extract and fraction was evaluated following the steps of Guideline 423, using female mice; LD(50) for both preparations was determined as 2,500 mg/kg body weight. CE and F80 promoted a reduction of the leukocyte number and nitrite level in inflammatory exudates when the anti-inflammatory assay (carrageenan-induced peritonitis) was performed. CE and F80 inhibited writhing regarding antinociceptive activity (acetic acid-induced writhing response in mice). In conclusion, CE and F80 have no significant cytotoxic or antitumor activities in cell lines showing low toxicity and no action against tumors in vivo. Both preparations revealed anti-inflammatory and antinociceptive activities, corroborating the pharmacological basis of L. ferrea for ethnomedical use.

Detection of water soluble lectin and antioxidant component from Moringa oleifera seeds.

Seed flour from Moringa oleifera is widely used as a natural coagulant for water treatment in developing countries. Extracts obtained by water soaking of M. oleifera intact seeds were investigated for the presence of lectin, trypsin inhibitor, tannin as well as antioxidant activity. A water soluble M. oleifera lectin (WSMoL) detected was mainly active with rabbit cells at pH 4.5; heat treatment, pH 7.0, fructose and porcine thyroglobulin abolished HA of WSMoL. Trypsin inhibitor or tannins were not detected; the antioxidant component (WSMoAC) reduced 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) was slower than catechin and was thermostable. The extracts showed a primary glycopolypeptide band of Mw 20,000; the main native acidic protein showed hemagglutinating activity. WSMoL may be involved in seed coagulant properties.

Antioxidant activity of leaf extracts from Bauhinia monandra.

Bauhinia monandra Kurz. is used in Brazil for the treatment of diabetes. Since this activity may be correlated with the presence of antioxidant compounds, leaf extracts of B. monandra were evaluated for their radical scavenging capacity (RSC). An ethanolic extract was taken up in aqueous methanol and partitioned with hexane, chloroform, ethyl acetate to yield three organic extracts together with remaining aqueous extract. The RSC was determined spectrophotometrically using 1,1-diphenylpicrylhydrazyl free radical (DPPH). The chloroform and ethyl acetate extracts were the most appropriate as sources of antioxidant compounds as shown by their inhibition concentration (IC50) and inhibition percentage (IP) values. The antioxidant activity of such extracts was attributed to the presence of three compounds of different polarities (flavonoids and steroids). The chloroform and ethyl acetate extracts exhibited an IC50 of approximately 2 mg/g DPPH and IP values in the range of 60-65%. The results indicate that the extracts of B. monandra have a very potent antioxidant activity, compared with the pure catechins used as positive controls and with other plant extracts.

Effect of arbuscular mycorrhizal fungi and soil phosphorus level on expression of protein and activity of peroxidase on passion fruit roots

The effects of mycorrhizal inoculation and increasing soil P levels on the expression of total proteins and peroxidase activity on passion fruit roots were evaluated. The experimental design was entirely at random, with four treatments of inoculation (a - control; b - Gigaspora albida; c - Scutellospora heterogama; d - mixture of G. albida, G. margarita, S. heterogama, and Glomus clarum) Ãù three levels of soil P (4, 11, and 30 mg/dm of soil), each with three replicates. Plants were harvested 70 days after inoculation, when root colonization, shoot P level, protein content, and enzymatic activity of peroxidase (PAGE - 7 percent) on root extract were evaluated. Regarding protein, there was no significant difference among the treatments, except between those roots receiving mixed inoculum and 11 mg P/dm of soil. Effect of P on protein concentration, when compared with the inoculation effect was observed. For peroxidase, there was an eletrophoretic band common to all treatments (rf: 0.43) and another that was absent only in noncolonized plants, grown in soil with lower P (rf: 0.46). Mycorrhizal specific bands were not present but a small decrease of intensity of bands in noncolonized plants was observed. Conversely, the control roots presented a single band (rf: 0.33) not observed in the other extracts, that may demonstrate an inhibitory effect of AMF on some host activities. The data showed the influence of P level in soil on the protein expression of roots, suggesting the influence of this nutrient on root genetic expression as well as on the mechanisms of symbiotic control/recognition