Antioxidant and antifungal activities of marrubiin, extracts and essential oil from Marrubium vulgare L. against pathogenic dermatophyte strains.

OBJECTIVE: Medicinal plants extracts and plant-derived compounds are one of the natural sources for discovering new antifungal agents, the objectives of this work were to investigate for the first time the antidermatophytic, antipathogenic activities of methanol, acetone extracts, and essential oil of Marrubium vulgare L. grown in Tunisia and its active compound marrubiin on pathogenic for animals and humans, such as some dermatophytes and pathogenic for plants, and to evaluate antioxidant activities of different extracts with consideration to their chemical compositions. MATERIAL AND METHODS: Acetone and methanol extracts were evaluated by HPLC, the essential oil was also analyzed by GC/MS. PCL assay was used to determine the antioxidant activity. RESULTS: Results showed that methanol and acetone extracts exhibited a significant antioxidant activity (261.41 and 272.90µmol TE/g respectively), while the lowest one was observed in the case of marrubiin and essential oil. The antifungal activity of different extracts, marrubiin and essential oil at two concentrations (20 and 100µg/mL) were screened against the dermatophytes fungi Microsporum gypseum, Microsporum canis, Arthroderma cajetani, Trichophyton mentagrophytes, Trichophyton tonsurans, Epidermophyton floccosum and against two fungi strains (Botrytis cinerea, Pythium ultimum). Among tested extracts, marrubiin at 100µg/mL showed about 50% inhibition for T. mentagrophytes and E. floccosum. The anti-phytopathogenic activity was also carried out, only marrubiin had in activity against B. cinerea at the highest dose (32.40%), while methanol extract of M.vulgare and marrubiin are able to increase the mycelial growth of P. ultimum at the highest concentration (45.15 and 40.30% respectively). CONCLUSION: In our study, we conclude that M.vulgare and marrubiin can be used as natural antioxidants and antifungal agent for treatment of skin dermatophyte infections.

Antifungal activity of 40 TCMs used individually and in combination for treatment of superficial fungal infections.

ETHNOPHARMACOLOGICAL RELEVANCE: A series of 40 important Traditional Chinese Medicines (TCMs), which were reported effective in treating superficial fungal infections of the skin in Chinese clinical trial publications and Chinese Herbal Classics, were chosen for the investigation of the individual and combination antifungal properties against 8 superficial fungal strains in vitro. MATERIALS AND METHODS: Plant preparations were followed the theory of TCM by using sterile water boiled with plant material at 100°C to produce water decoction of the tested sample. The minimum inhibitory concentration (MIC) of each plant for each fungus was determined. For the compatibility investigation, both invariable (same amounts of each tested TCM) and variable (different amounts of each tested TCM) combinations were evaluated. RESULTS: All the tested TCMs demonstrated varying degrees of antifungal activities against one or more of the tested superficial fungi, and 16 of which were effective on all of the fungi. Strong antifungal activities were exhibited by water decoction of 7 TCMs with MIC at about 100µg/ml, and among these effective antifungal extracts, 4 TCMs including Melaphis chinensis, Polygonum cuspidatum, Punica granatum and Schisandra chinensis showed the significantly inhibitory activities against all of the fungi with MICs among 50µg/ml. Most of the invariable combinations of the above-mentioned 4 TCMs showed synergic effects against 4 of the least susceptible fungi strains, especially the invariable combination of Punica granatum, Melaphis chinensis and Schisandra chinensis, with the MIC at 23.4µg/ml. However, their further variable combinations investigation demonstrated that only the combination of 7.5g Punica granatum with 10g Melaphis chinensis and 7.5g Schisandra chinensis showed synergic effect with the MIC at19.5µg/ml. CONCLUSIONS: The present study aimed the discovery of therapeutically useful agents for treatment of superficial fungal infections. Findings suggested that the combination of 3 TCMs including Punica granatum, Melaphis chinensis and Schisandra chinensis showed potential antifungal activity and thus appeared to be promising agents in preventing superficial fungal skin infectious in a natural way through herbal resources. The synergic effects of invariable and variable combinations of the tested TCMs threw a light on our further animal model and clinical practice as well as the bio-guided isolation and identification of the antifungal compounds.

The anti-dermatophyte activity of Allium hirtifolium Boiss aqueous extract.

In an attempt at demonstrating the efficacy of Allium hirtifolium aqueous extract in control of skin fungal infections as traditional use, we evaluated the anti-dermatophyte activities of A. hirtifolium aqueous extract from bulbs and of ketoconazole against Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, M. gypseum, Trichophyton schoenleinii and Trichophyton verrucosum var. album by food poisoning technique, disc diffusion and micro broth dilution assays. The anti-fungal activity of A. hirtifolium was excellent when it was compared with ketoconazole. The anti-fungal evaluation by food poisoning method showed that A. hirtifolium extract inhibited the growth of dermatophytes dose-dependently. The inhibition zone diameter (IZ) of A. hirtifolium extract (15 µg/disc) was in the range of 28.8 ± 0.31 to 67.7 ± 1.5mm, while ketoconazole (15 µg/disc) had the IZ lower than 13mm. The MIC and MFC values of A. hirtifolium extract were in the range of 0.2-1.7 and 0.4-0.7 µg/mL; respectively. Therefore, A. hirtifolium extract showed a strong anti-fungal activity against human and animal dermatophytes.

Antifungal activity of Aegle marmelos (L.) Correa (Rutaceae) leaf extract on dermatophytes.

OBJECTIVE: To evaluate the in vitro antifungal activity of Aegle marmelos leaf extracts and fractions on the clinical isolates of dermatophytic fungi like Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. METHODS: The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of various extracts and fractions of the leaves of Aegle marmelos were measured using method of National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Aegle marmelos leaf extracts and fractions were found to have fungicidal activity against various clinical isolates of dermatophytic fungi. The MIC and MFC was found to be high in water and ethyl alcohol extracts and methanol fractions (200µg/mL) against dermatophytic fungi studied. CONCLUSIONS: Aegle marmelos leaf extracts significantly inhibites the growth of all dermatophytic fungi studied. If this activity is confirmed by in vivo studies and if the compound is isolated and identified, it could be a remedy for dermatophytosis.

Antibacterial and antifungal activity of Cassia fistula L.: an ethnomedicinal plant.

Hexane, chloroform, ethyl acetate, methanol and water extracts from the flower of Cassia fistula (an ethnomedicinal plant) were tested against bacteria and fungi. All the extracts exhibited antibacterial activity against Gram-positive organisms with minimum inhibitory concentrations (MIC) between 0.078 and 2.5 mg/ml. Among the Gram-negative bacteria, only Pseudomonas aeruginosa was susceptible to the extracts. Ethyl acetate crude extract was fractionated using chromatographic techniques. A crystal was isolated, which was confirmed as 4-hydroxy benzoic acid hydrate using X-ray crystallography. It exhibited antifungal activity against Trichophyton mentagrophytes (MIC 0.5 mg/ml) and Epidermophyton floccosum (MIC 0.5 mg/ml).

Occurrence of dermatomycoses and in-vitro therapeutic efficacy of three antifungal drugs on the growth of Epidermophyton floccosum.

The occurrence of dermatomycoses and the in-vitro therapeutic efficacy of some antifungal agents on dermatomycotic organisms were investigated. Of the 550 primary school children screened, the incidence was one hundred (18%), 70 were males (representing 20% of the males screened) and 30 females (15% of the females sampled). The differences between male and female prevalence were insignificant. Three species of dermatophytes were isolated and identified. These were Microsporum canis, Trichophyton tonsurans and Epidermophyton floccosum. The antifungal agents tested on E. floccosum were griseofulvin, terbinafine and ketoconazole. They produced different sized zones of inhibition against the growth of E. floccosum. Griseofulvin exhibited a 50% inhibition of growth on E. floccosum at 63.00 mg/L. Terbinafine on the other hand exhibited varying levels of inhibition of growth at varying concentrations, at 0.07 mg/L, terbinafine achieved 46% inhibition of growth on E. floccosum. The drug achieved 100% inhibition of growth on the isolate at 61.81 mg/L. In the case of ketoconazole, 50% inhibition of growth was achieved at 100 mg/L while 100% inhibition of growth was achieved at 200 mg/L. The antifungal effects of the three drugs were confirmed by broth dilution tests where terbinafine was found to be fungistatic on the growth of E. floccosum at concentrations ranging from 0.013-1.700 mg/L and was fungicidal at concentrations ranging from 0.027-1.700 mg/L. Ketoconazole was found to inhibit the growth of E. floccosum at 0.003-1.700 mg/L and was fungicidal at concentrations ranging from 0.027-1.700 mg/L. It however did not succeed in killing the isolate under the same range of concentrations. Griseofulvin exhibited fungistatic effects on the growth of E. floccusum at 0.013-1.700 mg/L. In conclusion, ketoconazole and griseofulvin were found to be fungistatic and not fungicidal while terbinafine was both fungistatic and fungicidal on the pathogen. Terbinafine was found to be the most effective drug in inhibiting E. floccosum.

Clinical comparison of the efficacy and tolerability of once daily Canesten with twice daily Nizoral (clotrimazole 1% cream vs. ketoconazole 2% cream) during a 28-day topical treatment of interdigital tinea pedis.

The effects of two topical cream formulations containing clotrimazole 1% and ketoconazole 2%, respectively, were clinically compared in a double-blind, randomized manner for a 28-day therapy of interdigital tinea pedis in 106 treated patients. Ketoconazole was to be used twice daily whereas clotrimazole was administered only once daily. The primary response criterion defined as the number of patients with cure or improvement after 28 treatment days was comparable with 62.0% vs. 64.0% (clotrimazole vs. ketoconazole) for the full analysis set of 100 (50 vs. 50) patients. The mycological response revealed a negative culture and microscopy in 53.1% vs. 52.1% of the patients after 14, in 76.0% vs. 79.2% after 28, and in 83.7% vs. 76.9% after 56 days of observation, indicating a possibly better long-term efficacy of clotrimazole. The development of the overall score of tinea-related signs and symptoms did not show relevant differences between the two drugs and continuously decreased from 11+/-5 in both groups at baseline to 2+/-2 vs. 2+/-1 at day 56. As to the remission and improvement rates of single symptoms, better results were obtained under clotrimazole than under ketoconazole particularly for pruritus (97.8 vs. 89.6%) and burning/stinging (97.5 vs. 89.4%) which both are perceived as most bothersome by the patients. Furthermore, both substances appeared as comparably safe and well tolerable (8 vs. 7 adverse events with only 1 vs. 3 drug related). In conclusion, a successful therapy of tinea pedis can be achieved with both clotrimazole and ketoconazole within 28 days of treatment and once-daily clotrimazole is equally effective as twice-daily ketoconazole with favourable influences on the most irritating symptoms of the disease. Mycological and reliable clinical cure cannot be observed during two weeks after start of treatment.

Antimicrobial activity of protoanemonin, a lactone from ranunculaceous plants.

Protoanemonin, a component of Ranunculus bulbosus, was tested as an antifungal agent on selected strains of dermatophytes and yeasts. The minimum inhibitory concentrations ranged from 2.0 to 7.5 X 10(-4) M and the minimum lethal concentrations from 3.8 X 10(-4) M to greater than 1.0 X 10(-3) M. The most sensitive dermatophyte tested was Epidermophyton floccosum, and the most sensitive yeast Rhodotorula glutinis. The effects of different culture media and of light on the sensitivity of Rhodotorula glutinis to protoanemonin were also tested. Structural analogies between protoanemonin and other cytotoxic unsaturated lactones, and the reversal by the amino acid cysteine of the antifungal action suggest a possible mechanism of action.

The formation of macroconidia in dermatophytes following UV irradiation and temperature treatment.

The ability of sterile hyphae of Epidermophyton and Microsporum to produce macroconidia was restored by exposing hyphae to ultra violet irradiation followed by prolonged incubation at 35 degrees C. The four species of Trichophyton used in the present study did not produce macroconidia after such treatments.