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1.
J Mycol Med ; 30(1): 100927, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31983544

ABSTRACT

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.


Subject(s)
Antifungal Agents/pharmacology , Antioxidants/pharmacology , Arthrodermataceae/drug effects , Diterpenes/pharmacology , Marrubium/chemistry , Animals , Antifungal Agents/isolation & purification , Antioxidants/isolation & purification , Arthrodermataceae/classification , Arthrodermataceae/pathogenicity , Dermatomycoses/drug therapy , Dermatomycoses/microbiology , Diterpenes/isolation & purification , Epidermophyton/drug effects , Epidermophyton/growth & development , Humans , Microbial Sensitivity Tests , Microsporum/drug effects , Microsporum/growth & development , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Oils/chemistry , Plant Oils/pharmacology , Trichophyton/drug effects , Trichophyton/growth & development
2.
Mycoses ; 59(7): 416-8, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27144472

ABSTRACT

In general, it is recommended to incubate dermatophytes cultures for a minimum of 4 weeks. Several aspects of routine fungal cultures should be evaluated in order to implement appropriate and necessary changes. The aim of this study was to determine the optimum incubation time for routine dermatophytes cultures, analysing the time to find first fungal growth by visual observation. We recorded the time when the initial growth was detected for all dermatophyte isolates during a 4-year period. A total of 5459 dermatophyte cultures were submitted to our laboratory. From the total cultures, only 16 (1.42%) isolates were recovered over/after 17 days of incubation and only three dermatophyte species were recovered over 17 days. Fourteen isolates belong to Trichophyton rubrum, one isolate to Trichophyton mentagrophytes complex and one isolate to Epidermophyton floccosum. We concluded that an incubation period of 17 days is enough to establish a microbiological diagnosis of dermatophytosis.


Subject(s)
Arthrodermataceae/growth & development , Arthrodermataceae/isolation & purification , Epidermophyton/growth & development , Epidermophyton/isolation & purification , Mycology/methods , Time Factors , Tinea/diagnosis , Tinea/microbiology , Trichophyton/growth & development , Trichophyton/isolation & purification
3.
J Ethnopharmacol ; 163: 88-93, 2015 Apr 02.
Article in English | MEDLINE | ID: mdl-25625353

ABSTRACT

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.


Subject(s)
Antifungal Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Epidermophyton/drug effects , Microsporum/drug effects , Trichophyton/drug effects , Antifungal Agents/therapeutic use , Dermatomycoses/drug therapy , Drugs, Chinese Herbal/therapeutic use , Epidermophyton/growth & development , Medicine, Chinese Traditional , Microbial Sensitivity Tests , Microsporum/growth & development , Trichophyton/growth & development
4.
J Mycol Med ; 25(1): e10-4, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25456419

ABSTRACT

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.


Subject(s)
Allium/chemistry , Antifungal Agents/isolation & purification , Antifungal Agents/therapeutic use , Plant Extracts/therapeutic use , Tinea/drug therapy , Arthrodermataceae/drug effects , Epidermophyton/drug effects , Epidermophyton/growth & development , Foodborne Diseases/drug therapy , Foodborne Diseases/microbiology , Humans , Ketoconazole/pharmacology , Microbial Sensitivity Tests , Microsporum/drug effects , Microsporum/growth & development , Phytotherapy , Plant Extracts/pharmacology , Tinea/microbiology , Trichophyton/drug effects , Trichophyton/growth & development , Water/chemistry
6.
J Mycol Med ; 24(2): e57-64, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24746720

ABSTRACT

OBJECTIVE: The increasing importance of dermatophytoses and emerging resistance of dermatophytes to current synthetic antifungals have stimulated the search for safer and more effective alternative drugs from natural sources. The present study was carried out to identify antagonistic bacteria of soil origin with strong inhibitory activities on the growth of major human pathogenic dermatophytes. MATERIALS AND METHODS: Antifungal activity of isolated soil bacteria was screened against the dermatophytes from three genera Microsporum (M. canis, M. gypseum), Epidermophyton (E. floccosum) and Trichophyton (T. mentagrophytes, T. rubrum, T. violaceum, T. tonsurans) by using visual plate agar assay method. A Pseudomonas chlororaphis isolate S105, identified at the species level by 16S ribosomal RNA sequence analysis, was reported as the strongest antagonistic bacterium. P. chlororaphis S105 culture supernatant (PCCS) was examined against tested dermatophytes by GY (glucose-yeast extract) broth bioassay in 6-well microplates. Antifungal compound of the bacterium was partially purified from the culture supernatant through a purification scheme of methanol extraction, Diaion HP20 ion-exchange chromatography and preparative thin layer chromatography. RESULTS: P. chlororaphis S105 was the most potent inhibitor of fungal growth for all tested dermatophytes with a percent inhibition ranged from 57.1% to 99.8%. The PCCS suppressed the growth of all fungi tested in the range of 18.5% to 84.8%. Partially purified antifungal compound of the bacterium was identified as a phenazine-like compound with an Rf value of 0.51. The compound inhibited fungal growth by 73.6% to 97.9% on GY broth. Fungal growth inhibition was significant for all dermatophytes tested in comparison with the controls (Anova, P<0.05). CONCLUSION: With respect to the strong inhibitory activity of P. chlororaphis against pathogenic dermatophytes reported here, it may be considered as a rich source of useful metabolites with potential application in antifungal drug discovery.


Subject(s)
Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Pseudomonas/chemistry , Antibiosis , Antifungal Agents/analysis , Epidermophyton/drug effects , Epidermophyton/growth & development , Microbial Sensitivity Tests , Microsporum/drug effects , Microsporum/growth & development , Phenazines/isolation & purification , Phenazines/metabolism , Phenazines/pharmacology , Pseudomonas/isolation & purification , Pseudomonas/physiology , Soil Microbiology , Trichophyton/drug effects , Trichophyton/growth & development
7.
Int J Dermatol ; 52(12): 1557-60, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24134487

ABSTRACT

BACKGROUND: In this study, we aimed to establish the in vitro antifungal susceptibilities of terbinafine, miconazole, itraconazole, ketoconazole, griseofulvin, and amphotericin B against dermatophyte isolates. METHODS: One hundred and seventy-seven clinical isolates were tested: Trichophyton rubrum (n = 78), Trichophyton mentagrophytes (n = 49), Epidermophyton floccosum (n = 30), Trichophyton verrucosum (n = 16), and Trichophyton tonsurans (n = 4). The broth microdilution assay for antifungal susceptibility testing of dermatophytes was performed according to Clinical and Laboratory Standards Institute guidelines in the M38-A2 document. RESULTS: Our minimum inhibitory concentration (MIC) results showed that the values for terbinafine for all dermatophyte isolates were significantly lower than the values for amphotericin B, miconazole, itraconazole, ketoconazole, and griseofulvin. For T. rubrum isolates, amphotericin B was more active than miconazole, itraconazole, and ketoconazole. Among the antifungal drugs tested, griseofulvin had the highest minimum inhibitory concentration values for T. mentagrophytes isolates. CONCLUSION: Terbinafine was found to be the most effective antifungal drug against all tested dermatophyte isolates. Griseofulvin was the less active antifungal drug against T. mentagrophytes isolates. Performing antifungal susceptibility testing is especially important for screening the development of antifungal resistance.


Subject(s)
Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Arthrodermataceae/growth & development , Drug Resistance, Fungal , Itraconazole/pharmacology , Amphotericin B/pharmacology , Epidermophyton/drug effects , Epidermophyton/growth & development , Griseofulvin/pharmacology , Humans , In Vitro Techniques , Ketoconazole/pharmacology , Microbial Sensitivity Tests , Naphthalenes/pharmacology , Terbinafine , Trichophyton/drug effects , Trichophyton/growth & development , Turkey
8.
Rev Iberoam Micol ; 30(2): 130-3, 2013.
Article in Spanish | MEDLINE | ID: mdl-23318162

ABSTRACT

BACKGROUND: Dermatophytes can be divided into geophilic (soil), zoophilic (animals) and anthropophilic (humans) strains, depending on the source of the keratin that they use for nutritional purposes. AIMS: The in vitro susceptibility of clinical isolates of dermatophyte fungi has been studied in the 3 ecological groups with several antifungal agents for the topical management of dermatophytoses in order to determine their relationship with the ecological group. METHODS: A standardised dilution micromethod in a liquid medium was used for the determination of the in vitro antifungal activity of 9 topical antifungal drugs: amorolfine (AMR), bifonazole (BFZ), clotrimazole, econazole, ketoconazole, miconazole, oxiconazole, terbinafine (TRB) and tioconazole. The in vitro activity was obtained against 124 clinical isolates of dermatophyte moulds from the anthropophilic, zoophilic and geophilic ecological groups. RESULTS: The in vitro antifungal activity was different depending on the ecological group, although a species-dependent profile was also observed. CONCLUSIONS: Azole derivatives showed a similar antifungal profile, being more active against anthropophilic dermatophytes > zoophilic > geophilic. Activity of TRB and AMR was different from that of azole derivatives (zoophilic > anthropophilic > geophilic). A higher in vitro antifungal activity against the 3 ecological groups was observed with TRB and AMR, whilst BFZ was the less active drug.


Subject(s)
Antifungal Agents/pharmacology , Drug Resistance, Fungal , Epidermophyton/drug effects , Host Specificity , Microsporum/drug effects , Trichophyton/drug effects , Animal Diseases/microbiology , Animals , Antifungal Agents/classification , Aspergillus fumigatus/drug effects , Candida/drug effects , Drug Resistance, Multiple, Fungal , Epidermophyton/classification , Epidermophyton/growth & development , Humans , In Vitro Techniques , Microbial Sensitivity Tests , Microsporum/classification , Microsporum/growth & development , Mycoses/microbiology , Mycoses/veterinary , Soil Microbiology , Species Specificity , Trichophyton/classification , Trichophyton/growth & development
9.
Bioeng Bugs ; 3(2): 104-13, 2012.
Article in English | MEDLINE | ID: mdl-22539027

ABSTRACT

A total of 220 lactic acid bacteria isolates were screened for antifungal activity using Aspergillus fumigatus and Aspergillus niger as the target strains. Four Lactobacillus strains exhibited strong inhibitory activity on agar surfaces. All four were also identified as having strong inhibitory activity against the human pathogenic fungi Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. One of the four lactobacilli, namely Lb. reuteri ee1p exhibited the most inhibition against dermatophytes. Cell-free culture supernatants of Lb. reuteri ee1p and of the non-antifungal Lb. reuteri M13 were freeze-dried and used to access and compare antifungal activity in agar plate assays and microtiter plate assays. Addition of the Lb. reuteri ee1p freeze-dried cell-free supernatant powder into the agar medium at concentrations greater than 2% inhibited all fungal colony growth. Addition of the powder at 5% to liquid cultures caused complete inhibition of fungal growth on the basis of turbidity. Freeze-dried supernatant of the non-antifungal Lb. reuteri M13 at the same concentrations had a much lesser effect. As Lb. reuteri M13 is very similar to the antifungal strain ee1p in terms of growth rate and final pH in liquid culture, and as it has little antifungal activity, it is clear that other antifungal compounds must be specifically produced (or produced at higher levels) by the anti-dermatophyte strain Lb. reuteri ee1p. Reuterin was undetectable in all four antifungal strains. The cell free supernatant of Lb. reuteri ee1p was analyzed by LC-FTMS using an Accela LC coupled to an LTQ Orbitrap XL mass spectrometer. The high mass accuracy spectrum produced by compounds in the Lb. reuteri ee1p strain was compared with both a multianalyte chromatogram and individual spectra of standard anti-fungal compounds, which are known to be produced by lactic acid bacteria. Ten antifungal metabolites were detected.


Subject(s)
Antifungal Agents/pharmacology , Epidermophyton/growth & development , Lactobacillus/metabolism , Microsporum/growth & development , Animals , Antibiosis/physiology , Arthrodermataceae/growth & development , Glyceraldehyde/analogs & derivatives , Glyceraldehyde/pharmacology , Humans , Microbial Sensitivity Tests , Propane/pharmacology
10.
Med Mycol ; 50(6): 579-84, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22332906

ABSTRACT

Previous studies have described some antibacterial effects of antimicrobial peptides (AMPs) expressed in human skin, but little is known about their possible activity against dermatophytes. Therefore we have tested the effects of human ß-defensin 2 (hBD-2), ribonuclease 7 (RNase 7) and psoriasin on the in vitro growth of four dermatophyte species. Germinating conidia of Trichophyton rubrum, T. mentagrophytes, Microsporum canis and Epidermophyton floccosum were exposed in vitro to hBD-2, RNase 7, psoriasin and fluconazole. Subsequent fungal growth was measured photometrically over 168 hours. All AMPs significantly inhibited fungal growth, with the degree of inhibition dependent on the dermatophyte species and the specific AMP. E. floccosum was found to be the most susceptible species in that it was markedly suppressed by all AMPs, whereas M. canis was inhibited only by psoriasin. Overall, psoriasin was the most effective AMP and had even stronger inhibitory effects on some dermatophytes than fluconazole. Our findings show that AMPs expressed in human skin can, in principal, inhibit the growth of dermatophytes in vitro. Therefore the question whether AMPs are relevant for human protection against tineas is justified and should be addressed by investigating their role in vivo.


Subject(s)
Arthrodermataceae/drug effects , Ribonucleases/pharmacology , S100 Proteins/pharmacology , Trichophyton/drug effects , beta-Defensins/pharmacology , Antifungal Agents/pharmacology , Epidermophyton/drug effects , Epidermophyton/growth & development , Epidermophyton/isolation & purification , Female , Fluconazole/pharmacology , Humans , Male , Microbial Sensitivity Tests , Microsporum/drug effects , Microsporum/growth & development , Microsporum/isolation & purification , S100 Calcium Binding Protein A7 , Spores, Fungal/drug effects , Spores, Fungal/growth & development , Tinea Pedis/microbiology , Trichophyton/growth & development , Trichophyton/isolation & purification
11.
Asian Pac J Trop Biomed ; 1(4): 309-12, 2011 Aug.
Article in English | MEDLINE | ID: mdl-23569781

ABSTRACT

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.


Subject(s)
Aegle/chemistry , Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Plant Extracts/pharmacology , Plant Leaves/chemistry , Antifungal Agents/isolation & purification , Arthrodermataceae/growth & development , Epidermophyton/drug effects , Epidermophyton/growth & development , Humans , Microbial Sensitivity Tests , Microbial Viability/drug effects , Microsporum/drug effects , Microsporum/growth & development , Plant Extracts/isolation & purification , Trichophyton/drug effects , Trichophyton/growth & development
12.
J Ethnopharmacol ; 112(3): 590-4, 2007 Jul 25.
Article in English | MEDLINE | ID: mdl-17532583

ABSTRACT

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).


Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Cassia/chemistry , Plant Extracts/pharmacology , Acetates/chemistry , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Chloroform/chemistry , Crystallization , Epidermophyton/drug effects , Epidermophyton/growth & development , Ethnopharmacology , Flowers/chemistry , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/growth & development , Hexanes/chemistry , Hydroxybenzoates/chemistry , Hydroxybenzoates/isolation & purification , Hydroxybenzoates/pharmacology , India , Methanol/chemistry , Microbial Sensitivity Tests , Molecular Structure , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plants, Medicinal/chemistry , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/growth & development , Trichophyton/drug effects , Trichophyton/growth & development , Water/chemistry
13.
Mycoses ; 48(6): 442-6, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16262884

ABSTRACT

Anti-dermatophytic activity of Chrysosporium keratinophillum against species of the genera Trichophyton, Microsporum and Epidermophyton floccosum was tested in vitro. When C. keratinophillum and different species of dermatophytes were inoculated on Sabouraud's dextrose agar plates 2 cm apart, no antagonistic effect of C. keratinophillum on the mycelial growth of dermatophytes was observed. However, conidia production was not observed on the hyphae of Trichophyton rubrum, Trichophyton tonsurans and E. floccosum grown near C. keratinophillum. The secretory substances released by C. keratinophillum inhibited the growth of T. rubrum, T. tonsurans, Trichophyton mentagrophytes var. interdigitale and E. floccosum at a concentration of 2,000 microg ml(-1) when tested by broth dilution technique. No inhibition of the growth was observed for Microsporum gypseum and Microsporum nanum. The anti-fungal activity of secretory substances released by C. keratinophillum was recorded to be heat stable. Results of the present study suggest that the anti-dermatophytic activity of the secretory substances of C. keratinophillum on T. rubrum, T. mentagrophytes var. interdigitale, T. tonsurans and E. floccosum may be responsible in part, for the absence of these dermatophyte species in soil. Considering the global prevalence of C. keratinophillum in soil one may speculate that the anti-dermatophytic activity of C. keratinophillum is one of the early events for the evolutionary divergence of saprophytic archi-dermatophytes to obligate parasitic dermatophyte species.


Subject(s)
Antibiosis , Chrysosporium/physiology , Epidermophyton/growth & development , Microsporum/growth & development , Trichophyton/growth & development , Antifungal Agents/isolation & purification , Antifungal Agents/pharmacology , Biological Evolution , Chrysosporium/metabolism , Hot Temperature , Mycelium/growth & development , Soil Microbiology , Spores, Bacterial
14.
Med Mycol ; 43(4): 319-25, 2005 Jun.
Article in English | MEDLINE | ID: mdl-16110777

ABSTRACT

The introduction of systemic antifungal drugs which act upon different targets is the main issue of the in vivo antifungal resistance control. Different factors, such as growth curve phase, quality of the specimen, quantity of the inoculum, temperature, pH, culture medium composition, incubation duration and solvent, are believed important factors affecting minimum inhibitory concentration (MIC) value to most of the antifungal agents. We assayed an in vitro susceptibility test with 40 isolates of dermatophytes: Microsporum canis, Trichophyton rubrum, Trichophyton mentagrophytes and Epidermophyton floccosum against griseofulvin, fluconazole, itraconazole and terbinafine, using the guidelines of the M38-P document approved by the NCCLS. We determined the growth curves, to estimate the specific growth rate (mu max) and the generation time (G) of each dermatophyte, using dry weight and spectrophotometry methods. We demonstrate that, at 192 h, all fungi tested had a constant growth curve and we considered this as the optimal time for MIC determination. Terbinafine, griseofulvin and itraconazole possessed the highest antifungal activity against the four groups of dermatophytes studied. Fluconazole demonstrated no efficacy. Our MIC results differ from other authors and this difference is due to the timing of the MIC determination based on the growth curve of each fungi tested.


Subject(s)
Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Arthrodermataceae/growth & development , Drug Resistance, Fungal , Epidermophyton/drug effects , Epidermophyton/growth & development , Microbial Sensitivity Tests/methods , Microbial Sensitivity Tests/standards , Microsporum/drug effects , Microsporum/growth & development , Trichophyton/classification , Trichophyton/drug effects , Trichophyton/growth & development
15.
Mycopathologia ; 156(4): 295-301, 2003.
Article in English | MEDLINE | ID: mdl-14682454

ABSTRACT

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.


Subject(s)
Antifungal Agents/pharmacology , Epidermophyton/drug effects , Tinea/drug therapy , Antifungal Agents/therapeutic use , Child , Epidermophyton/growth & development , Epidermophyton/metabolism , Female , Griseofulvin/pharmacology , Griseofulvin/therapeutic use , Humans , Ketoconazole/pharmacology , Ketoconazole/therapeutic use , Male , Microbial Sensitivity Tests , Naphthalenes/pharmacology , Naphthalenes/therapeutic use , Nigeria/epidemiology , Prevalence , Terbinafine , Tinea/epidemiology , Tinea/microbiology
16.
Mycoses ; 46(3-4): 120-5, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12870200

ABSTRACT

The aim of this study was to investigate a possible effect of optical brighteners on the growth of dermatophytes. Typical strains of Trichophyton rubrum, T. mentagrophytes, Microsporum canis and Epidermophyton floccosum were grown on agar plates containing two different brighteners of stilbenedisulfonic acid type in concentrations between 5 x 10(-5) and 1 x 10(-2) mol l-1 and their thallus diameters were compared with controls. In addition, hyphae grown with brighteners were compared with controls by fluorescence microscopy and by transmission electron microscopy. Both brighteners had a significant dose-dependent growth-suppressive effect on all dermatophytes tested, that was complete at a concentration of 10(-2) and 10(-3) mol l-1, respectively. Fluorescence microscopy of hyphae showed a pronounced fluorescence of the septal areas and a less-intense staining of the outer cell walls. Electron microscopy revealed a marked thickening and blurred contours of the cell walls grown with brighteners. These new observations relate very well to an interference of optical brighteners with the formation of normal chitin fibrils as described previously. Optical brighteners of stilbenedisulfonic acid type may be rewarding objects for the development of new antifungal agents.


Subject(s)
Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Arthrodermataceae/growth & development , Stilbenes/pharmacology , Chitin/metabolism , Dose-Response Relationship, Drug , Epidermophyton/drug effects , Epidermophyton/growth & development , Fluorescence , Humans , Microbial Sensitivity Tests , Microscopy, Electron , Microscopy, Fluorescence , Microsporum/drug effects , Microsporum/growth & development , Stilbenes/chemistry , Trichophyton/drug effects , Trichophyton/growth & development , Ultraviolet Rays
17.
Fungal Genet Biol ; 36(1): 47-57, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12051894

ABSTRACT

The antifungal activity of 3-methyl-5-aminoisoxazole-4-thiocyanate, a new azole derivative, was studied on the dermatophyte Epidermophyton floccosum. The compound strongly inhibited the in vitro growth of two different strains of the fungus and even induced profound morphogenetic anomalies. Optical and electron microscopy showed that such treatment targets the endomembrane system, particularly the plasmalemma, causing abnormal extrusion of the wall mannans. This results in improper arrangement of the different parietal materials; the walls are thus weak and subject to subapical rupture which terminates cell growth and elongation of the hypha. The morphological results and the preliminary biochemical data on fungal sterols suggest that this compound employs an action mechanism similar to that of other azoles used in therapy.


Subject(s)
Antifungal Agents/pharmacology , Epidermophyton/drug effects , Epidermophyton/metabolism , Isoxazoles/pharmacology , Mannans/metabolism , Thiocyanates/pharmacology , Cell Wall/drug effects , Cell Wall/metabolism , Cell Wall/ultrastructure , Epidermophyton/growth & development , Epidermophyton/ultrastructure , Microscopy, Electron , Microscopy, Electron, Scanning
18.
Mycoses ; 45(3-4): 91-6, 2002 Apr.
Article in English | MEDLINE | ID: mdl-12000508

ABSTRACT

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.


Subject(s)
Antifungal Agents/administration & dosage , Antifungal Agents/therapeutic use , Clotrimazole/administration & dosage , Clotrimazole/therapeutic use , Ketoconazole/administration & dosage , Ketoconazole/therapeutic use , Tinea Pedis/drug therapy , Adolescent , Adult , Double-Blind Method , Drug Administration Schedule , Drug Therapy, Combination , Epidermophyton/drug effects , Epidermophyton/growth & development , Humans , Middle Aged , Ointments/administration & dosage , Ointments/therapeutic use , Tinea Pedis/microbiology
20.
Microbiol Immunol ; 41(5): 377-86, 1997.
Article in English | MEDLINE | ID: mdl-9194036

ABSTRACT

An ATP bioluminescence assay as a rapid reference method for fluconazole (FLCZ) susceptibility testing of dermatophytes, as well as yeasts, was developed and evaluated by comparing it with viability, turbidity and fungal protein content-based conventional methods. FLCZ susceptibility results obtained with strains of Candida albicans and dermatophytes by the bioluminescence method in high-resolution medium were well correlated with those obtained by conventional methods currently used in clinical microbiology laboratories or reported previously, including a broth dilution method by the National Committee for Clinical Laboratory Standards (NCCLS). Thus, ATP bioluminescence assay can be used to monitor fungal growth in liquid culture media. The procedure has considerable potential for the rapid testing of FLCZ susceptibility of dermatophytes and other fungi.


Subject(s)
Adenosine Triphosphate/metabolism , Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Firefly Luciferin , Fluconazole/pharmacology , Microbial Sensitivity Tests/methods , Arthrodermataceae/growth & development , Candida albicans/drug effects , Candida albicans/growth & development , Epidermophyton/drug effects , Epidermophyton/growth & development , Luciferases , Luminescent Measurements , Microsporum/drug effects , Microsporum/growth & development , Time Factors , Trichophyton/drug effects , Trichophyton/growth & development
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