Bacteriostatic effect of Echeveria extracts on diarrheagenic E. coli pathotypes and non-cytotoxicity on human Caco-2 cells.

INTRODUCTION: Diarrheagenic Escherichia coli pathotypes are important aetiological agents of diarrhoeal illness among children from less developed areas, worldwide. Diarrheagenic E. coli pathotypes strains are increasingly becoming drug resistant, thus effective and accessible therapeutic alternatives are required for their treatment; herbal extracts may be a potential alternative. AIMS: to evaluate Echeveria craigiana, E. kimnachii, and E. subrigida methanol extracts antibacterial effect on six diarrheagenic E. coli reference strains and on human colorectal adenocarcinoma cells viability and cytokine production. METHODOLOGY: Diarrheagenic E. coli pathotypes reference strains: typical enteropathogenic E2348/69, enterotoxigenic H10407, enterohaemorrhagic O157:H7/EDL933, enteroinvasive E11, diffusely adherent C18451-A, and enteroaggregative 042 E. coli. E craigiana, E. kimnachii, and E. subrigida leaves, collected at Sinaloa, Mexico, were freeze-dried and macerated in methanol solvent. Antibacterial activity was determined by a novel method developed in our laboratory, bacterial oxygen consumption by polarographic oxygen electrode technique and membrane integrity by two methods (live/dead and protein leakage assays). Colorectal adenocarcinoma cells viability by MTT assay and cytokine production using a Cytometric Bead Array kit. RESULTS: Extracts concentrations of 100 µg/mL and 5-hour incubation, reduced more than 93% the growth of all diarrheagenic E. coli pathotypes tested strains and significantly decreased bacterial oxygen consumption, like bacteriostatic antibiotics. After 24-hour incubation methanol extracts had a differential antibacterial effect on each diarrheagenic E. coli pathotypes strain. Echeveria extracts did not have any effect on viability and cytokine production of colorectal adenocarcinoma cells. CONCLUSIONS: Echeveria methanol extracts have a bacteriostatic effect on all diarrheagenic E. coli pathotypes strains, thus potentially they could be used as antibacterial agents on diarrheagenic E. coli pathotypes-contaminated products and on patients with diarrheagenic E. coli pathotypes infections.

Bioguided isolation of N-malonyl-(+)-tryptophan from the fruit of Pithecellobium dulce (Roxb.) Benth. that showed high activity against Hymenolepis nana.

Pithecellobium dulce is distributed in America and Asia where is widely used in traditional medicine. This study describes the bioguided fractionation of the methanol extract (ME) obtained from the P. dulce fruit that showed in vitro activity against Hymenolepis nana; Artemia salina assay was used to determine toxicity; and the purified compound was computationally analysed to obtain its absorption-distribution-metabolism-excretion-and-toxicity properties (ADMET). The ME and its fractions were more active than praziquantel (PZQ), and the purified compound was characterized as N-malonyl-(+)-tryptophan (NMT). Parasites treated with NMT showed shorter paralysis and death times (5 and 7 min) than those treated with PZQ (15 and 30 min), both used at 20 mg/mL. Toxicity and ADMET prediction results supported the slight-hazardousness and efficacy of the assayed fractions/compound. This is the first report of the antiparasitary activity of both the P. dulce ME and NMT, showing their potential to treat human H. nana infections.

Soluble melanins of the Randia echinocarpa fruit - Structural characteristics and toxicity.

The water-soluble melanins (SM) of Randia echinocarpa fruit possess interesting biological activities and have been scarcely characterized. In this study, SM were obtained at boiling (SMBT) and room (SMRT) temperatures and characterized by UV-Vis, IR, thermogravimetric analysis, and GC-MS of the hydrolysis products of the SM; besides, the solid-state 13 C NMR, elemental analysis, and acute and sub-acute toxicity of the SMBT were determined. SMBT and SMRT contain organic acids and carbohydrates and their spectroscopic signals and thermograms were similar, but the SMBT yield was higher. The SMBT were characterized by their elemental composition (C 48.260 ± 0.011%, N 3.693 ± 0.009% and H 6.093 ± 0.076%) consistent with the presence of aromatic rings and eumelanins, degradation temperature at 300°C, 13 C NMR signals supporting melanin-bonding with carbohydrates and organic acids, and innocuity in Balb/C mice (acute assay, LD50  > 5 g/kg b.w.; sub-acute assay, no lethality at 500 mg/kg b.w. for 30 days). PRACTICAL APPLICATIONS: The consumption of melanins has been associated with health benefits because of their biological activities (e.g., antioxidant, immunostimulatory, UV- and radiation-protective). Randia echinocarpa is employed in Mexican traditional medicine against chronic degenerative diseases (e.g., cancer and diabetes) and ailments of organs (e.g., kidney and lung) and systems (e.g., circulatory and gastrointestinal). The R. echinocarpa fruit contains water-soluble melanins (SM) that inhibit carbohydrate-digestive enzymes and show high antioxidant activity; thus, SM could be useful for the prevention and treatment of diabetes. This study showed that the SM structure contains melanin-bonding organic acids and carbohydrates, which could be associated with the SM solubility and higher yield, and that SMBT were innocuous in the acute and sub-acute assays in mice. Thus, the R. echinocarpa SMBT could be used as safe potential ingredients to develop functional products.

High prevalence of multidrug-resistant Escherichia coli isolates from children with and without diarrhoea and their susceptibility to the antibacterial activity of extracts/fractions of fruits native to Mexico.

PURPOSE: This paper aims to evaluate the antimicrobial resistance of Esherichia coli isolates from children under 5 years old, with and without diarrhoea, who were hospital outpatients in Culiacan, Sinaloa, Mexico. It also looks at the antimicrobial activity of fruit extracts against selected multidrug-resistant (MDR) E. coli strains. METHODOLOGY: A total of 205 E. coli isolates from stool samples were collected from 94 children under 5 years old who were outpatients from two hospitals in the city of Culiacan, Sinaloa, Mexico, during the autumn/winter of 2003/04; their resistance profiles to 19 commercial antimicrobials were investigated using the Kirby-Bauer method. The antibacterial activities of extracts/fractions of fruits (i.e. uvalama, Vitex mollis; ayale, Crescentia alata; and arrayan, Psidium sartorianum) were evaluated using the broth microdilution method. RESULTS: All E. coli isolates were susceptible to amikacin, nitrofurantoin and meropenem, and approximately 96 % were resistant to at least one antimicrobial, especially carbenicillin (93.2 %), cefuroxime sodium (53.7 %), ampicillin (40 %) and trimethoprim/sulfamethoxazole (35.1 %). Likewise, the frequency of MDR strains (44.9 %) was high, and no significant association with diarrhoea symptoms was found. Remarkably, all fruit extracts/fractions showed antibacterial activity against some, but not all, MDR isolates. The lowest minimal inhibitory concentration values were for the hexane fraction of arrayan (0.25 mg ml-1). CONCLUSION: A high number of antimicrobial-resistant E. coli (especially to ß-lactams and sulfonamides) and MDR isolates were detected in children under 5 years old, irrespective of diarrhoea symptoms; this is novel information for Culiacan, Sinaloa, Mexico. Moreover, our results showed that the studied fruit extracts/fractions are potential alternative or complementary treatments for MDR E. coli strains.

Chemical composition and biological activities of Helicteres vegae and Heliopsis sinaloensis.

CONTEXT: Helicteres vegae Cristóbal (Sterculiaceae) (Hv) and Heliopsis sinaloensis B.L. Turner (Asteraceae) (Hs) are endangered and poorly studied plant species; related plants have been used against chronic-degenerative and infectious diseases. Therefore, Hv and Hs could be sources of bioactive compounds against these illnesses. OBJECTIVE: To determine the chemical composition and biological activities (antioxidant, antimutagenic and antimicrobial) of Hv and Hs leaves (L) and stems (S). MATERIALS AND METHODS: Methanol extracts (ME) of each plant/tissue were evaluated for their phytochemicals; phenolics (HPLC-DAD-ESI-MS); antioxidant activity (AA) (0.125-4 mg/mL) (DPPH, ABTS, ORAC and ß-carotene discoloration); antimutagenicity (0.5 and 1 mg/plate) (Ames assay, tester strain Salmonella enterica serovar Typhimurium YG1024, 1-nitropyrene as mutagen); activity against human pathogens (1 mg/mL); and toxicity (0.01-2 mg/mL) (Artemia salina assay). RESULTS: All ME showed flavonoids and triterpenes/steroids. The ME-SHv had the highest content of total phenolics (TP) (2245.82 ± 21.45 mg GAE/100 g d.w.) and condensed tannins (603.71 ± 1.115 mg CE/100 g d.w.). The compounds identified were flavonoids (kaempferol 7-O-coumaroylhexoside, and two kaempferol 7-O-rhamnosylhexosides) and phenolics [rosmarinic acid, and 3'-O-(8″-Z-caffeoyl) rosmarinic acid]. The ME-LHs showed the highest content of flavonoids (357.88 mg RE/g d.w.) and phenolic acids (238.58 mg CAE/g d.w.) by HPLC. The ME-SHv showed the highest AA. All ME were strong antimutagens (63.3-85.7%). Only the Hs extracts were toxic (ME-LHs, LC50 = 94.9 ± 1.7 µg/mL; ME-SHs, LC50 = 89.03 ± 4.42 µg/mL). DISCUSSION AND CONCLUSIONS: Both Hv and Hs are potential sources of preventive and therapeutic agents against chronic-degenerative diseases.

Actividad antibacteriana de extractos de frutos de nanchi (Byrsonima crassifolia (L.) Kunth), arrayán (Psidium sartorianum (O. Berg) Nied.) y ayale (Crescentia alata Kunth) / Antibacterial activity of extracts obtained from the nanchi (Byrsonima crassifolia (L.) Kunth), arrayan (Psidium sartorianum (O. Berg) Nied.) and ayale (Crescentia alata Kunth) fruits

Nanchi (Byrsonima crassifolia), arrayan (Psidium sartorianum) and ayale (Crescentia alata) are wild and under-utilized plants from Mexico; their fruits have been used as food and as Mexican traditional remedies against human bacterial infections (e.g. bacillary dysentery). However, scientific reports which support such uses or promote their consumption are scarce. In this work, the antibacterial activities of fruit extracts (i.e. hexanic, HE; chloroformic, CE; and methanolic, ME) were determined by the micro-dilution assay, establishing the Minimum Inhibitory Concentration (MIC) and Minimum Bactericide Concentration (MBC) against 21 human pathogenic bacteria. The HE of arrayan and ayale showed the highest activity against enterobacteria (E. coli, Salmonella spp. and Shigella spp.) (MIC 0.25-2 mg/mL; MBC 0.5-16 mg/mL). The arrayan ME was the most active against the Gram-positive bacteria, showing Staphylococcus aureus the highest sensitivity (MIC 2 mg/mL; MBC 2-4 mg/mL). The presented results support the traditional uses of these plant materials for treating bacterial infectious diseases.

Nanchi (Byrsonima crassifolia), arrayán (Psidium sartorianum) y ayale (Crescentia alata) son plantas silvestres subutilizadas de México; sus frutos son comestibles y usados como medicamentos tradicionales contra infecciones bacterianas humanas (e.g. disentería bacilar). Sin embargo, los reportes científicos que avalen los usos y promuevan su consumo son escasos. En este trabajo se determinó, ensayo de micro-dilución en caldo, la Concentración Mínima Inhibitoria (CMI) y Concentración Mínima Bactericida (CMB), de los extractos de frutos (hexánico, EH; clorofórmico, EC; y metanólico, EM) contra 21 bacterias patógenas humanas. Los EH de arrayán y ayale mostraron la mayor actividad (CMI 0.25-2 mg/mL; CMB 0.5-16 mg/mL) contra enterobacterias (Escherichia coli, Salmonella spp. y Shigella spp.). El EM de arrayán fue el más activo contra bacterias Gram positivas, presentando Staphylococcus aureus la mayor sensibilidad (CMI 2 mg/mL; CMB 2-4 mg/mL). Estos resultados apoyan el uso tradicional de estos materiales en padecimientos asociados al tratamiento de infecciones bacterianas.

Physicochemical, nutritional and antibacterial characteristics of the fruit of Bromelia pinguin L.

Bromelia pinguin L. is a plant native to Sinaloa, Mexico, where its fruit is used as food or as a phytotherapeutic agent. The fruits of B. pinguin were characterized and they could be considered as a functional food. These fruits show an average weight of 13.7 g and a yellow color of high luminosity (b* = 43.2, L* = 74.5). The values for acidity (4.6%, as citric acid) and pH (3.7) of B. pinguin fruit are similar to those of citrics. The edible portion is characterized by a high content of vitamin C (126 mg/100 g), ash (10.6 g/100 g d.w.), crude fiber (3.4 g/100 g d.w.), calcium (1290 mg/100 g d.w.), magnesium (500 mg/100 g d.w.), manganese (2.95 mg/100 g d.w.) and it is a good source of zinc (2.8 mg/100 g d.w.). Polar fractions extracted from the pulp fruit showed activity against several genera of human pathogenic-bacteria (Staphylococcus, Streptococcus, Enterococcus, Salmonella, Shigella, Escherichia and Pseudomonas).