Down-regulatory effect of Thymus vulgaris L. on growth and Tri4 gene expression in Fusarium oxysporum strains.

The aims of this study were to evaluate the efficacy of Thymus vulgaris (T. vulgaris) essential oil on the fungal growth and Tri4 gene expression in Fusarium oxysporum (F. oxysporum) strains. The oil was obtained by water-distillation using a Clevenger-type system. The chemical composition of the essential oil was obtained by gas chromatography- mass spectroscopy (GC-MS) and by retention indices. The antifungal activity was evaluated by broth microdilution assay. A quantitative real time RT-PCR (qRT-PCR) assay was also developed specific for F. oxysporum on the basis of trichothecene biosynthetic gene, Tri4, which allowed discrimination from F. oxysporum. Results showed thymol (32.67%) and p-cymene (16.68%) as the main components of T. vulgaris. Minimum inhibitory concentration (MIC) values varied from 5 to 20 µg/ml with T. vulgaris (mean: 10.50 µg/ml), while minimum fungicidal concentration (MFC) values ranged from 8 to 30 µg/ml with mean value of 16.20 µg/ml qRT-PCR results revealed a downregulation from 4.04 to 6.27 fold of Tri4 gene expression of the fungi exposed to T. vulgaris essential oil. The results suggest that T. vulgaris oil can be considered potential alternative natural fungicide to the synthetic chemicals that are currently used to prevent and control seed-borne diseases.

Chemical composition and antifungal activity of Satureja hortensis L. essentiall oil against planktonic and biofilm growth of Candida albicans isolates from buccal lesions of HIV(+) individuals.

ETHNOPHARMACOLOGICAL RELEVANCE: Oral candidiasis is an opportunistic infection of the oral cavity which usually occurs in the immunocompromised individuals. Candida albicans (C. albicans) is the most common species of yeast responsible for oral candidiasis. This study investigated the effects of Satureja hortensis L. essentiall oil (EO) on the planktonic, biofilm formation and mature biofilms of C. albicans isolates from buccal lesions of HIV(+) individuals. MATERIALS AND METHODS: MTT reduction assay, broth micro-dilution method and scanning electron microscopy (SEM) were employed to determine the effect of mentioned EO on the C. albicans planktonic and biofilm forms. GC-GC/MS was used to detect the major active compounds of EO. RESULTS: Thymol (45.9%), gamma-terpinen (16.71%), carvacrol (12.81%) and p-cymene (9.61%) were found as the most abundant constituents. MIC values ranged from 250 to 400 µg/ml and MFC values ranged from 350 to 500 µg/ml. All C. albicans isolates formed biofilm on polystyrene plats but the quantity of biofilm mass (optical density) was different for the isolates ranging from 0.850 to 0.559 nm. The mean of biofilm formation by C. albicans isolates was reduced by 87.1 ± 3.7%, 73.6 ± 5.1%, 69.4 ± 5.3% and 67 ± 4.2% at 4800, 3200, 2400 and 1600 µg/ml, respectively. In sub-MIC concentration, SEM analysis revealed loosening of cells, deformity of three dimensional structures of biofilms and shrinkage in cell membranes of sessile cells. CONCLUSIONS: In conclusion, the substantial anti-fungal activity showed by S. hortensis L. EO suggests exploitation of this oil as potential natural anti-biofilm product to deal with the problem of buccal cavity lesion associated with C. albicans.

Yeasts as important agents of onychomycosis: in vitro activity of propolis against yeasts isolated from patients with nail infection.

OBJECTIVE: The purposes of this study were to determine the frequency of the yeast species obtained from patients with clinical features of onychomycosis and the in vitro antifungal susceptibility of the yeast species to propolis. METHODS: A prospective study was carried out at the Mycology Research Center in Iran from 2010 to 2011. Clinical diagnosis was performed by direct microscopic examination and culture. Different yeast species were identified by morphological and biochemical tests. An antifungal susceptibility test to fluconazole (FLU) and propolis by the broth microdilution method was performed on each isolate. RESULTS: One hundred and twenty-eight fungal isolates were obtained. The most prevalent fungi were yeasts (81, 63.2%), dermatophytes (36, 28.1%), and nondermatophyte fungi (11, 8.6%). Fingernails were more affected than toenails (65.4% vs. 19.8%, respectively). The most frequently found species was Candida albicans (38.5%), followed by Candida spp. (23.1%), C. tropicalis (10.8%), C. kefyr (6.2%), C. krusei (3.1%), Malassezia globosa (4.6%), M. slooffiae (4.6%), and M. pachydermatis (1.5%). Of all yeast isolates (65), seven showed resistance to FLU. The average MIC of propolis for FLU-susceptible isolates was 5.8 µg/mL, whereas this value was 12.25 µg/mL for FLU-resistant isolates. CONCLUSION: Our results proved that the propolis inhibits the growth of pathogenic yeasts and confirmed the efficiency of propolis as an anti-Candida and anti-Malassezia agent.

Evaluation of the antifungal activity of Zataria multiflora, Geranium herbarium, and Eucalyptus camaldolensis essential oils on Saprolegnia parasitica-infected rainbow trout (Oncorhynchus mykiss) eggs.

The purpose of the present study was to evaluate and assess the capability of Zataria multiflora, Geranium herbarium, and Eucalyptus camaldolensis essential oils in treating Saprolegnia parasitica-infected rainbow (Oncorhynchus mykiss) trout eggs. A total of 150 infected eggs were collected and plated on glucose-pepton agar at 24°C for 2 weeks. The antifungal assay of essential oils against S. parasitica was determined by a macrodilution broth technique. The eggs were treated with essential oils at concentrations of 1, 5, 10, 25, 50, and 100 ppm daily with three repetitions until the eyed eggs stage. Of 150 eggs examined, S. parasitica (54.3%), Saprolegnia spp. (45%), and Fusarium solani (0.7%) were isolated. The minimum inhibitory concentrations of Z. multiflora, E. camaldolensis, and G. herbarium essential oils against S. parasitica were 0.9, 2.3, and 4.8 ppm, respectively. Zataria multiflora and E. camaldolensis at concentrations of 25, 50, and 100 ppm, and G. herbarium at concentration of 100 ppm had significant differences in comparison with negative control (p<0.05). The results revealed that malachite green, followed by Z. multiflora, E. camaldolensis, and G. herbarium treated eggs had remained the most number of final eyed eggs after treatment. The highest final larvae rates belonged to malachite green, E. camaldolensis, Z. multiflora, and G. herbarium, respectively. The most hatching rates were recorded with malachite green (22%), and then Z. multiflora (11%), E. camaldolensis (7%), G. herbarium (3%), and negative control (1%). Zataria multiflora and E. camaldolensis were more effective than G. herbarium for the treatment of S. parasitica-infected rainbow trout eggs in aquaculture environment.

Inhibition of aflatoxin production and growth of Aspergillus parasiticus by Cuminum cyminum, Ziziphora clinopodioides, and Nigella sativa essential oils.

Aflatoxins are highly toxic and carcinogenic metabolites produced by Aspergillus parasiticus on food and agricultural commodities. Natural products may control the production of aflatoxins. The aims of this study were to evaluate the effects of the essential oils (EOs) of Cuminum cyminum, Ziziphora clinopodioides, and Nigella sativa on growth and aflatoxins production by A. parasiticus. Minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of the EOs were determined and compared with each other. Determination of aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)) was performed by immunoaffinity column extraction using reverse phase-high performance liquid chromatography. The major oil components were α-pinene (30%) in C. cyminum, pulegone (37%) in Z. clinopodioides, and trans-anthol (38.9%) in N. sativa oils. In broth microdilution method, C. cyminum oil exhibited the strongest activity (MIC(90): 1.6; MFC: 3.5 mg/mL), followed by Z. clinopodioides (MIC(90): 2.1; MFC: 5.5 mg/mL) and N. sativa (MIC(90): 2.75; MFC: 6.25 mg/mL) oils against A. parasiticus (p<0.05). Aflatoxin production was inhibited at 0.25 mg/mL of C. cyminum and Z. clinopodioides oils, of which that of C. cyminum was a stronger inhibitor. C. cyminum EO caused significant reductions in values of 94.2% for AFB(1), 100% for AFB(2), 98.9% for AFG(1), 100% for AFG(2), and 97.5% for total aflatoxin. It is concluded that the EOs of C. cyminum, Z. clinopodioides, and N. sativa could be used as natural inhibitors in foods at low concentrations to protect from fungal and toxin contaminations by A. parasiticus.