Interaction of carvacroland voriconazole against drug - resistant Candida strains isolated from patients with candidiasis.

OBJECTIVE: Terpenoid phenols, especially 5-isopropyl-2-methylphenol (carvacrol), are components of plant essential oils that exhibit potent antifungal activity against a wide range of pathogens, including Candida species. The purpose of this study was to determine the interaction activity of carvacrol in combination with voriconazole against Candida albicans (C. albicans), C. glabrata and C. krusei isolates. MATERIALS AND METHODS: The minimum inhibitory and fungicidal concentrations (MICs and MFCs) of carvacrol and voriconazole were determined against various Candida species isolated from patients with candidiasis using the Clinical Laboratory Standards Institute (CLSI) M27-A2 broth microdilution method. The nature of the interaction was studied from fractional inhibitory concentration indices (FICIs) for carvacrol plus voriconazole combination calculated from checkerboard microdilution assay. RESULTS: Carvacrol presented an antifungal effect, with mean MICs of 66.87µg/mL for C. albicans, 75µg/mL for C. glabrata and 95µg/mL for C. krusei isolates. The mean MICs of voriconazole against C. albicans, C. glabrata and C. krusei isolates were 0.087, 1.25 and 0.35µg/mL, respectively. Carvacrol in combination with voriconazole exhibited the synergistic anti-Candida effects against all species of Candida tested. FICI values for carvacrol plus voriconazole combination ranged from 0.370 to 0.853 for C. albicans isolates, 0.412 to 0.625 for C. glabrata isolates, and 0.474 to 0.748 for C. krusei isolates. No antagonistic activity was seen in the strains tested. CONCLUSION: From these results we suggest that carvacrol has great potential as antifungal, and that voriconazole can be supplemented with carvacrol to inhibit clinical Candida isolates.

Potential effect of 2-isopropyl-5-methylphenol (thymol) alone and in combination with fluconazole against clinical isolates of Candida albicans, C. glabrata and C. krusei.

Limitations of antifungals used in the treatment of candidiasis, as the development of resistant strains, are known by the scientific community. In this context, the aim of this study was to investigate the activity of 2-isopropyl-5-methylphenol (thymol) in combination with fluconazole (FLZ) against clinical Candida strains. The antifungal activity of thymol along with FLZ was evaluated by the Clinical Laboratory Standards Institute (CLSI) M27-A2 broth microdilution method. In addition, synergism was observed for clinical strains of Candida spp. with combination of thymol-FLZ evaluated by the chequerboard microdilution method. The mean of minimum inhibitory concentration (MIC) values of thymol and FLZ were 49.37 and 0.475µg/ml for C. albicans, 51.25 and 18.80µg/ml for C. glabrata and 70 and 179.20µg/ml for C. krusei strains, respectively. Thymol in combination with FLZ exhibited the synergistic effects against all species of Candida tested. FICI values for thymol plus FLZ ranged from 0.366 to 0.607 for C. albicans strains, 0.367 to 0.482 for C. glabrata strains, and 0.375 to 0.563 for C. krusei strains. No antagonistic activity was seen in the strains tested. Thymol was found to have a fungicidal effect on Candida species and a synergistic effect when combined with FLZ.

Evaluation of murine lung epithelial cells (TC-1 JHU-1) line to develop Th2-promoting cytokines IL-25/IL-33/TSLP and genes Tlr2/Tlr4 in response to Aspergillus fumigatus.

OBJECTIVE: The aims of this study were to determine the role of live and heat-killed Aspergillus fumigatus conidia in releasing interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP) and to express Toll-like receptor (Tlr)2 and Tlr4 genes. MATERIALS AND METHODS: Murine lung epithelial cells were incubated with live and heat-killed A. fumigatus conidia at 37°C for 6, 24 and 48h. After treatments, ELISA was performed to measure the concentrations of IL-25, IL-33 and TSLP in the supernatants. Quantitative real-time PCR (qPCR) was performed to assess the expression levels of Tlr2 and Tlr4 genes. RESULTS: The concentrations of IL-25 and IL-33 significantly increased after exposure to live and heat-killed conidia for various times when compared with untreated control (P<0.05). The secretion of TSLP at different concentrations of heat-killed conidia was significantly higher than both live conidia and untreated control (P<0.05). qRT-PCR results indicated a up-regulation from 1.08 to 3.60-fold for Tlr2 gene expression and 1.20 to 1.80-fold for Tlr4 gene expression exposed to heat-killed conidia. CONCLUSION: A. fumigatus has a potential ability to stimulate murine lung epithelial cells to produce IL-25/IL-33/TSLP, as well as to express Tlr2/Tlr4 genes, indicating an important role of lung epithelial cells in innate immune responses to A. fumigatus interaction.

Monitoring of mycoflora in outdoor air of different localities of Ahvaz, Iran.

INTRODUCTION: Airborne fungi are considered important causes of respiratory allergy and diseases. The knowledge of these fungi in a city or region is important for the ecological diagnosis and specific treatment of respiratory manifestations induced by inhalation of fungal agents. Therefore, in recent years bioaerosols and their density in the air is highly regarded. Identifying fungi with common mycological methods as well as molecular about 5% of people are prone to allergic symptoms of the respiratory tract caused by molds during their lives. Fungi are being considered as one of the most common triggers of asthma, allergic rhinitis and other respiratory problems. AIMS: We aimed at the investigation of the airborne fungi of Ahvaz, Khuzestan province, Iran, during spring and autumn months. MATERIALS AND METHODS: A total of 224 Petri dishes with Sabouraud dextrose agar medium containing chloramphenicol were exposed at seven different regions in the city. Two hundred and twenty-four samples (112 samples per season) were taken from outdoor air seven areas of Ahvaz using microbial sampler (Quick Take-30, SKC, USA) with Debbi 14.3L/min during 5minutes directly on Sabouraud dextrose agar medium with Chloramphenicol and Dechlorane. The effects of environmental factors such as temperature, humidity and wind velocity on bioaerosol concentrations were studied. RESULTS: The dishes exposed yielded 1240 fungal colonies of 24 genera. The most predominant were: Cladosporium spp. (37.1%), Alternaria spp. (25.7%), Aspergillus spp. (13.5%), Penicillium spp. (5.9%), Drechslera spp. (3.46%) and Stemphylium spp. (3.46%). The maximum and minimum numbers of airborne fungi were isolated from humid (32.3%) and industrial (17%) environments, respectively. In addition, positive correlation was observed between the number of fungi and relative humidity (r=0.476, P=0.000) in spring and autumn seasons. CONCLUSION: This study indicated that the incidence of airborne fungi with clinical significance had a direct relationship with the variation of environmental conditions.

Fungicidal efficacy of various honeys against fluconazole-resistant Candida species isolated from HIV+ patients with candidiasis.

OBJECTIVE: Honey is well known to possess a broad spectrum of activity against medically important organisms. The purpose of this study was to assess the antifungal activity of different honeys against 40 fluconazole (FLU) resistant Candida species, including Candida albicans (C. albicans), Candida glabrata, Candida krusei and Candida tropicalis. MATERIALS AND METHODS: Three honey samples were collected from northern (Mazandaran, A), southern (Hormozgan, B) and central (Lorestan, C) regions of Iran. A microdilution technique based on the CLSI, M27-A2 protocol was employed to compare the susceptibility of honeys "A", "B" and "C" against different pathogenic Candida isolates. RESULTS: The results showed that different Candida isolates were resistant to FLU, ranging from 64µg/mL to 512µg/mL. All of the honeys tested had antifungal activities against FLU-resistant Candida species, ranging from 20% to 56.25% (v/v) and 25% to 56.25% (v/v) for minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs), respectively. Honey "A" (MIC: 31.59%, v/v) showed higher anti-Candida activity than honey "B" (MIC: 35.99%, v/v) and honey "C" (MIC: 39.2%, v/v). No statistically significant differences were observed among the mean MIC values of the honey samples (P>0.05). The order of overall susceptibility of Candida species to honey samples were; C. krusei>C. glabrata>C. tropicalis>C. albicans (P>0.05). In addition, the mean MICs of Candida strains isolated from the nail, vagina and oral cavity were 33.68%, 36.44% and 39.89%, respectively, and were not significantly different (P>0.05). CONCLUSION: Overall, varying susceptibilities to the anti-Candida properties of different honeys were observed with four FLU-resistant species of Candida. Further research is needed to assess the efficacy of honey as an inhibitor of candidal growth in clinical trials.

Presence and distribution of yeasts in the reproductive tract in healthy female horses.

BACKGROUND: Yeasts are commensal organisms found in the reproductive and gastrointestinal tracts, and on the skin and other mucosa in mammals. OBJECTIVES: The purpose of this study was to isolate and identify yeast flora in the caudal reproductive tract in healthy female horses. STUDY DESIGN: Longitudinal study. METHODS: A total of 453 samples were collected using double-guarded swabs from the vestibule, clitoral fossa and vagina in 151 horses. All samples were cultured on Sabouraud 4% dextrose agar and incubated at 35°C for 7-10 days. Isolates were identified according to their morphological characteristics and biochemical profiles. RESULTS: Yeast colonies were isolated from 60 (39.7%) of the 151 horses. The isolated yeasts belonged to nine genera, and included Candida spp. (53.2%), Cryptococcus spp. (12.2%), Saccharomyces spp. (10.5%), Geotrichum spp. (8.0%), Rhodotorula spp. (7.1%), Malassezia spp. (3.7%), Trichosporon spp. (2.6%), Kluyveromyces spp. (2.6%) and Sporothrix spp. (0.2%). Candida krusei (43.1%) was the most frequent Candida species isolated. There was a significant difference in prevalence between C. krusei and other Candida species (P<0.05). The vestibule contained more yeast isolates (48.0%) than the vagina (18.3%). The isolation of yeast colonies from multiparous females (76.8%) was significantly higher than from maiden mares (P<0.05). MAIN LIMITATIONS: The study was limited by the difficulty of distinguishing between normal flora and potential pathogens. CONCLUSIONS: Candida spp., in particular C. krusei, represent important flora resident in the caudal reproductive tract in healthy female horses. This is particularly important in contexts that require the initiation of empirical treatment prior to the completion of culture results.

Antifungal effects of Lavandula binaludensis and Cuminum cyminum essential oils against Candida albicans strains isolated from patients with recurrent vulvovaginal candidiasis.

OBJECTIVE: Recurrent vulvovaginal candidiasis (RVVC), which affects approximately 5% of women of reproductive age, is defined as 4 or more episodes of symptomatic Candida vaginitis within a year. The purposes of this study were to determine the chemical compositions and antifungal susceptibility of Cuminum cyminum (C. cyminum) and Lavandula binaludensis (L. binaludensis) essential oils and their combination against Candida albicans (C. albicans) strains isolated from patients with RVVC. MATERIALS AND METHODS: C. albicans isolates were identified via germ tube test, CHROMagar and RapID Yeast Plus System. The essential oils were obtained by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography-mass spectroscopy (GC-MS). The broth microdilution method was used as antifungal susceptibility test (CLSI-M27-A3). RESULTS: The GC-MS analysis allowed 13 components to be determined; the main components of C. cyminum and L. binaludensis essential oils were γ-terpinene (21.07%) and 1,8-cineole (71.56%), respectively. L. binaludensis and C. cyminum oils were effective in inhibiting C. albicans growth at mean concentrations of 7.91±1.61µg/mL and 8.00±1.89µg/mL, respectively. In addition, the combination of C. cyminum with L. binaludensis oils were more active causing inhibition in all C. albicans isolates, with concentrations varying from 3.90 to 11.71µg/mL (mean value: 7.22±1.69µg/mL). CONCLUSION: The results suggested the potential substitution of the antifungal chemicals by C. cyminum and L. binaludensis essential oils as natural inhibitors to control the growth of the most important pathogenic Candida species and alternative therapies for RVVC.

In vitro antifungal activity of essential oils extracted from plants against fluconazole-susceptible and -resistant Candida albicans.

BACKGROUND AND PURPOSE: Candidaalbicans is the most common cause of candidal infections. Various studies have shown drug resistance among C. albicans isolates; thus, it is necessary to discover replacement treatments for Candida infections. In this study, we aimed to compare the effects of different essential oils against azoles-resistant and azoles-susceptible isolates. MATERIALS AND METHODS: Twenty fluconazole-resistant and 20 susceptible C. albicans isolates obtained from oral, vaginal, and cutaneous tissues of patients with candidiasis were evaluated. The efficacy and minimum inhibitory concentrations (MICs) of Zataria multiflora, Geranium herbarum,Lavendula officinalis,Cuminum, cyminum,Allium heamanthoides, and Artemisia sieberi essential oils against C. albicans were determined on the basis of a reference method for broth microdilution susceptibility testing of yeasts as suggested by Clinical and Laboratory Standards Institute (CLSI, M27-S4). After inoculation, incubation, and subculturation, the MICs were determined through comparison with the control. RESULTS: The obtained MICs for Zataria multiflora, Geranium herbarum, Artemisia sieberi, Allium heamanthoides, Cumminum cyminum, and Lavendula officinalis were 0.1-0.25 µl/ml (mean: 0.155 µl/ml), 0.625-1.66 µl/ml (mean: 0.93 µl/ml) 0.833-2.0 µl/ml (mean: 1.21 µl/ml), 0.1-0.25 µl/ml (mean: 0.155 µl/ml), 2-4 µl/ml (mean: 3.1 µl/ml), and 1.5-3.0 µl/ml (mean: 2.4 µl/ml), respectively. The results showed that Zataria multiflora and Allium heamanthoides essential oils were more efficient than other essential oils against Candida species. There were no significant differences between various Candida strains in terms of susceptibility to the essential oils. In addition, there were no significant differences in the MICs of these essential oils against the azoles-resistant and azoles-susceptible isolates. CONCLUSION: In this study, the anti-Candida effects of six essential oils against both azoles-resistant and azoles-susceptible isolates were similar. Given the documented resistance of different Candida species to synthetic and chemical antifungals, these essential oils are effective replacement treatments for cutaneous and mucosal Candida infections, especially in resistant or recurrent cases.

An epidemiological study of animals dermatomycoses in Iran.

OBJECTIVE: To determine the fungal species isolated from skin lesions of different animals suspected of having dermatomycoses and their prevalence in different regions of Iran. MATERIALS AND METHODS: A total of 1011 animals (292 dogs, 229 cats, 168 horses, 100 camels, 98 cows, 60 squirrels, 37 birds, 15 sheep, 6 goats, 5 rabbits and 1 fox) suspected of having dermatomycoses were examined. The samples were obtained by plucking the hairs and feathers with forceps around the affected area and scraping the epidermal scales with a sterile scalpel blade. All collected samples were analyzed by direct microscopy and culture. Laboratory identification of the fungal isolates was based on their colonial, microscopic and biochemical characteristics. RESULTS: Fungal agents were recovered from 553 (54.7%) animals suspected of having dermatomycoses. Of 553 confirmed cases, 255 (49.7%) were positive for dermatophytosis, 251 (45.4%) for Malassezia dermatitis, 14 (2.5%) for candidiasis, 12 (2.2%) for aspergillosis and 1 (0.2%) for zygomycosis. Cats (36.3%) were the most prevalent infected animals, followed by camels (13.4%), dogs (12.8%), horses (12.5%), cows (12.3%), squirrels (5.4%), birds (3.6%), sheep (2%), goats (1.1%), rabbits (0.4%) and fox (0.2%). Microsporum canis (M. canis) was the most frequent fungus isolated from dogs and fox, Malassezia pachydermatis (M. pachydermatis) from cats, horses and squirrels, Trichophyton verrucosum (T. verrucosum) from cows and camels, T. mentagrophytes var. mentagrophytes from sheep, goats and rabbits, and Aspergillus fumigatus (A. fumigatus) from birds. CONCLUSION: The results suggested that periodic screening of animals suspected of having dermatomycoses and necessary treatments could help in the management of their public health problem.

Purification and comparison of heat shock protein 90 (Hsp90) in Candida albicans isolates from Malaysian and Iranian patients and infected mice.

OBJECTIVE: The purposes of this study were to purify and compare the concentration ratios of heat shock protein 90 (Hsp90) in clinical isolates of Candida albicans (C. albicans) obtained from Malaysian and Iranian patients and infected mice. MATERIALS AND METHODS: Hsp90 was extracted using glass beads and ultracentrifugation from yeast cells and purified by ion exchange chromatography (DEAE-cellulose) and followed by affinity chromatography (hydroxyapatite). Purity of Hsp90 was controlled by SDS-PAGE and its identification was realized by immunoblotting test. RESULTS: The graphs of ion exchange and affinity chromatography showed one peak in all C. albicans isolates obtained from both Malaysian and Iranian samples, infected mice and under high-thermal (42°C) and low-thermal (25°C) shock. In immunoblotting, the location of Hsp90 fragments was obtained around 47, 75 and 82kDa. The least average concentration ratios of Hsp90 were 0.350 and 0.240mg/g for Malaysian and Iranian isolates at 25°C, respectively, while the highest average concentration ratios of Hsp90 were 3.05 and 2.600mg/g for Malaysian and Iranian isolates at 42°C, respectively. There were differences in the ratio amount of Hsp90 between Malaysian isolates (1.01±0.07mg/g) and mice kidneys (1.23±0.28mg/g) as well as between Iranian isolates (0.70±0.19mg/g) and mice kidneys (1.00±0.28mg/g) (P<0.05). CONCLUSION: The results showed differences in all situations tested including Iranian and Malaysian isolates, samples treated with temperatures (25°C or 42°C) and before and after infecting the mice (37°C), indicating higher virulent nature of this yeast species in high temperature in human and animal models.

Efficacy of medicinal essential oils against pathogenic Malassezia sp. isolates.

OBJECTIVES: The purposes of this study were to evaluate the distribution pattern and population size of Malassezia species in dogs with atopic dermatitis (AD) and the inhibitory efficacy of Zataria multiflora, Thymus kotschyanus, Mentha spicata, Artemisia sieberi, Rosmarinus officinalis and Heracleum persicum essential oils against pathogenic Malassezia isolates. METHODS: The samples were collected from 5 different anatomical sites of 33 atopic dogs and cultured onto modified Dixon agar (MDA) and Sabouraud dextrose agar (SDA) media. The essential oil extraction was performed by steam distillation using Clevenger system. Anti-Malassezia efficacy of medicinal essential oils and standard drugs was evaluated using broth microdilution method. RESULTS: A total of 103 yeast colonies were isolated from dogs with AD. Eight different Malassezia species were identified as follows: Malassezia pachydermatis (81.4%), M. globosa (7.8%), M. restricta (3.9%), M. sloofiae (2.9%), M. furfur (1%), M. nana (1%), M. obtusa (1%) and M. sympodialis (1%). The most and least infected sites were: anal (21.2%) and ear (10.6%) respectively. M. pachydermatis was the most frequent Malassezia species isolated from both skin and mucosa of dogs with AD. Antifungal susceptibility test revealed the inhibitory efficacy of essential oils on pathogenic Malassezia isolates with minimum inhibitory concentration (MIC(90)) values ranging from 30 to 850 µg/mL. Among the tested oils, Z. multiflora and T. kotschyanus exhibited the highest inhibitory effects (P<0.05). CONCLUSION: The essential oils of Z. multiflora and T. kotschyanus showed strong antifungal activity against pathogenic Malassezia species tested.

Evaluation of antioxidant and antifungal properties of the traditional plants against foodborne fungal pathogens.

OBJECTIVE: To determine the antioxidant and antifungal activities of the essential oils from five aromatic herbs, including Thymus vulgaris, Chamaemelum nobile, Ziziphora clinopodioides, Zingiber officinale and Cuminum cyminum, against different Aspergillus and Penicillium species. METHODS: The oils were subjected to screening for their possible antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The susceptibility test for the oils was carried out in terms of minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) using microdilution method. RESULTS: The values of the essential oils in DPPH assay were as follows: T. vulgaris (450.11±5.23 µg/mL), Ch. nobile (602.73±4.8 µg/mL), Ziz. clinopodioides (1238.82±9.3 µg/mL), Cu. cyminum (1255.52±8.92 µg/mL) and Zin. officinale (5595.06±8.24 µg/mL). Our findings also indicated a strong activity against tested fungi for the oil of T. vulgaris (1250 µg/mL), followed by Cu. cyminum (1416 µg/mL), Zin. officinale (1833 µg/mL), Ziz. clinopodioides (2166 µg/mL) and Ch. nobile (3750 µg/mL). This study confirmed the excellent antifungal and antioxidant properties of the essential oils, especially T. vulgaris, against foodborne pathogenic fungi. CONCLUSION: Owing to their strong protective features, these oils could be used in ethno-medicine as preventers of lipid peroxidation and cellular damage, and in food industries as preservers of foodstuffs against spoilage fungi. Also, they could be the candidates to develop new antibiotics and disinfectants to control infective agents.

Lactic acid bacteria as functional probiotic isolates for inhibiting the growth of Aspergillus flavus, A. parasiticus, A. niger and Penicillium chrysogenum.

OBJECTIVE: The aim of this study was to assess the potential of lactic acid bacteria (LAB) such as Lactobacillus acidophilus, L. rhamnosus, L. casei, L. paracasei and Bifidobacterium bifidum to inhibit the outgrowth of some common food-spoiling fungi including Aspergillus niger, A. flavus, A. parasiticus and Penicillium chrysogenum. METHODS: Bacterial isolates were cultured on Mann Rogosa Sharpe (MRS) broth and liquid cultures and supernatants were prepared. The antifungal activity was tested using the agar well diffusion method. RESULTS: Both liquid culture and supernatant of L. casei isolate exhibited high antifungal activity, followed by L. acidophilus and L. paracasei isolates. The least activity was recorded for the isolates B. bifidum, while the isolate L. rhamnosus was moderately active against tested fungi. The antifungal activity of the supernatants obtained from all probiotic isolates against fungi was significantly less than that of liquid cultures (P<0.05). Antifungal activity evaluation showed that A. flavus was the most inhibited fungus by probiotic bacteria, followed by P. chrysogenum, A. niger and A. parasiticus. CONCLUSION: These results suggest that probiotic bacteria strains have the ability to prevent the growth of pathogenic and mycotoxigenic fungi as antifungal agents for various biomedical applications.

Antifungal effect of Trachyspermum ammi against susceptible and fluconazole-resistant strains of Candida albicans.

OBJECTIVE: Trachyspermum ammi (T. ammi) has been known as having many therapeutic properties and its antimicrobial activity has currently received a renewed interest. This study aimed to verify the effectiveness of T. ammi essential oil to inhibit the growth of Candida albicans (C. albicans) strains isolated from HIV(+) patients with oropharyngeal candidiasis (OPC). MATERIALS AND METHODS: The essential oil was obtained by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography. Susceptibility tests were expressed as inhibition zone by the disk diffusion method and minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) by the broth microdilution method. RESULTS: Thymol (63.4%), p-cymene (19%) and γ-terpinen (16.9%) were found as the most abundant constituents. The disk diffusion results revealed that 67% of oral C. albicans isolates were susceptible, 9% susceptible-dose dependent and 24% resistant to fluconazole. In the broth microdilution method, 68% of isolates were susceptible, 5% susceptible-dose dependent and 27% resistant to fluconazole. The increase in concentration led to a significant reduction in yeasts that were growing in exponential phase. In addition, with increasing in T. ammi oil concentration, the time of remaining cells in lag phase was significantly increased. CONCLUSION: This study showed that all clinical C. albicans isolates were susceptible to T. ammi essential oil, indicating a significant reduction in the yeast growth in exponential phase.

A study of onychomycosis in patients attending a dermatology center in Tehran, Iran.

OBJECTIVE: To determine the incidence of onychomycosis based on age and sex, morphological pattern of the disease, predisposing factors and identification of fungus by direct microscopy and culture methods. METHODS: A prospective study was conducted on 140 patients with nail disorders. A detailed history and thorough examination was done in all patients. The samples were taken from patients clinically suspected of fingernails and toenails infections attending a dermatology center in Tehran, Iran. The nails were subjected to potassium hydroxide (KOH) examination and fungal culture on Sabouraud's dextrose agar (SDA) medium. RESULTS: Specimens from 79 patients (56.4%) were positive for onychomycosis. The mycological observations showing positive fining with KOH were observed in 79 (56.4%) and culture positive in 35 (25%) cases. Females were more infected than males. The most common age group infected was 41-60 years (40.7%). Toenails were affected more frequently than fingernails and dystrophic onychomycosis was the most common clinical type seen in 39.2% patients. From the culture-positive samples, yeasts were the most common pathogens isolated from 25 (71.4%) patients, followed by non-dermatophytic moulds in 6 (17.1%) and dermatophytes in 4 (11.5%) patients. CONCLUSION: This study demonstrated that Candida species were the main agents causing onychomycosis in our region and accurate diagnosis of onychomycosis was based on direct microscopy and fungal culture.

Immunomodulatory efficacy of ethanol extract of propolis on tumor-bearing mice with disseminated candidiasis.

OBJECTIVE: This study was aimed at investigating the effect of propolis on immunosurveillance by measuring the levels of serum interleukin (IL)-4, IL-10, IL-17, tumor necrosis factor (TNF)-α and interferon (IFN)-γ in tumor-bearing mice with disseminated candidiasis. METHODS: The ethanol extract of propolis was selected for this study. Balb/C female mice were infected with Candida albicans (C. albicans) and inoculated with spontaneous mouse mammary tumor (SMMT). The serum levels of tissue inhibitor of metalloproteinase-1(TIMP-1) were assessed by enzyme- linked immunosorbent assay (ELISA). Mice were treated daily with propolis solution (100mg/kg, 0.1 mL, orally) for 3 days before IV challenge with C. albicans and SC challenge with SMMT and continued for 10 days. The rates of survival and tumor growth of understudy mice were investigated as well. The levels of TNF-α, IFN-γ, IL-4, IL-10 and IL-17 cytokines in culture supernatants were determined by ELISA. RESULTS: The mean tumor size was significantly increased in tumor-bearing mice infected with C. albicans (16.98 ± 0.49 mm(2)) as compared to other mice groups (P<0.05). The results showed a significant decline of IL-4 and IL-10 levels after propolis administration to tumor-bearing mice infected with C. albicans (53.41 pg/mL, 156.81 pg/mL and 63.45 pg/mL) (P < 0.05). The increment of TNF-α (433.85 pg/mL) and IFN-γ (120.43 pg/mL) levels were also observed. CONCLUSION: Data revealed that propolis has remarkable immunomodulatory effect, which provides a scientific validation for the popular use of this natural substance, and further investigation will help to understand propolis usefulness during immunosuppressive conditions.

Genotypic variation and antifungal susceptibly of Candida zeylanoides clinical isolates.

OBJECTIVE: To investigate the DNA fingerprinting of Candida zeylanoides (C. zeylanoides) strains and the correlation between genotyping and antifungal susceptibility of C. zeylanoides. METHODS: Paper discs containing nystatin, ketoconazole, fluconazole, Zataria multiflora (Z. multiflora) and Pulicaria gnaphalodes (P. gnaphalodes) essential oils were used in the disc diffusion method to evaluate the in vitro activity of the antifungal agents by measuring the mean diameter of inhibition around the discs. Yeast isolates were characterized by randomly amplified polymorphic DNA (RAPD) analysis using two different primers. RESULTS: The mean inhibition zones were calculated 36.1 ± 2.2mm for ketoconazole, 25.3 ± 2.0mm for nystatin and 14.9 ± 1.4mm for fluconazole. Z. multiflora essential oil revealed a 58.6 ± 2.6mm mean zone of inhibition while P. gnaphalodes showed a 36.7 ± 1.8mm zone of inhibition against all isolates tested. A total of 14 strains of C. zeylanoides were divided into three types, 1 was genotype A strain, 2 genotype B strains and 11 genotype C strains. Strain of genotype A was significantly more susceptible to ketoconazole and nystatin than fluconazole (P<0.05). Strains of genotype B were more susceptible to ketoconazole than other antifungal agents (P>0.05). Strains of genotype C were significantly more susceptible to ketoconazole than fluconazole and nystatin (P<0.05). There was a significant correlation between C. zeylanoides genotypes and antifungal susceptibility (P < 0.05). CONCLUSION: We concluded that all C. zeylanoides genotypes were susceptible to nystatin and Z. multiflora essential oil. The correlation between antifungal susceptibility and C. zeylanoides genotype may be of potential therapeutic significance and larger studies are needed to prove this finding.

Potential effects of Trachyspermum copticum essential oil and propolis alcoholic extract on Mep3 gene expression of Microsporum canis isolates.

OBJECTIVE: The purpose of this study was to evaluate the effects of Trachyspermum copticum (T. copticum) essential oil and propolis alcoholic extract on growth and transcription of Mep3 gene of Microsporum canis (M. canis) strains. METHODS: The antifungal activity was assayed by broth macrodilution method. Fungal isolates were grown in soy peptone liquid medium and treated with T. copticum oil and propolis extract. Total RNAs of M. canis were subjected to reverse transcription-polymerase chain reaction (RT-PCR). Specific primers of Actin and Mep3 genes were used. RESULTS: The results revealed that MIC values of T. copticum oil against M. canis strains were ranged from 0.2-30.5 µg/mL, with 42.3% of the strains inhibited at 0.9 µg/mL. In addition, MIC values of propolis extract against M. canis strains were ranged from 0.2-488.2 µg/mL, with 34.6% of the strains inhibited at 0.9 µg/mL. RT-PCR analysis of Mep3 and Actin expression showed DNA fragments of 661 and 690 bp amplified in all isolates before treatments with T. copticum essential oil and propolis extract. Both T. copticum and propolis completely inhibited the expression of Mep3 gene. CONCLUSION: We reported for the first time that T. copticum and propolis inhibits the expression of Mep3 gene in M. canis strains in relation to a remarkable inhibition in protease production by the fungus.

Antifungal efficacy of thymol, carvacrol, eugenol and menthol as alternative agents to control the growth of food-relevant fungi.

OBJECTIVE: This work is an attempt to examine the antifungal activity of thymol, carvacrol, eugenol and menthol against 11 food-decaying fungi. METHODS: The susceptibility test for the compounds was carried out in terms of minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) using microdilution method in 96 multi-well microtiter plates. RESULTS: Results indicated that all compounds were effective to varying extents against various fungal isolates, with the highest efficacy displayed by carvacrol (mean MIC value: 154.5 µg/mL) (P<0.05). The incorporation of increased concentrations of all compounds to the media led to progressive and significant reduction in growth for all fungi. The most potent inhibitory activity of thymol, carvacrol, eugenol and menthol was found for Cladosporium spp. (MIC: 100 µg/mL), Aspergillus spp. (MIC: 100 µg/mL), Cladosporium spp. (MIC: 350 µg/mL), and Aspergillus spp. and Cladosporium spp. (MIC: 125 µg/mL), respectively. CONCLUSION: Thus, the application of these herbal components could be considered as a good alternatives to inhibit fungal growth and to reduce the use of synthetic fungicides.

Altered immune responses in patients with chronic mucocutaneous candidiasis.

OBJECTIVE: The purpose of this study was to investigate the lymphocyte transformation responses and cytokine secretion of peripheral blood mononuclear cells (PBMC) from patients with chronic mucocutaneous candidiasis (CMC). METHODS: Phytohaemagglutinin (PHA) mitogen and Candida albicans (C. albicans) antigen proliferation assays were performed by culturing PBMCs in RPMI 1640. The levels of interleukin (IL)-2, IL-10, IL-17 and interferon (IFN)-γ present in the supernatant of cultures were determined using enzyme-linked immunosorbent assay (ELISA). RESULTS: The results showed that most patients (92.3%) had a low proliferative response to C. albicans antigens and PHA. PBMCs from CMC patients produced lower levels of T (h)-1 cytokines IL-2 (78.5±59.8 pg/mL) and IFN-γ (115.1±43.3 pg/mL) in response to Candida antigens when compared to controls (Il-2: 177±103.6 pg/mL; IFN-γ: 330.3±21.6 pg/mL) (P<0.05). Conversely, we observed a partial enhancement of IL-10 in the patients (213.7±86.1 pg/mL). Production of IL-17 indicated no significant differences between patients and controls when stimulated by Candida antigens (21.5±8.6 pg/mL versus 32.4±12.2 pg/mL) and PHA (27.7±11.5 pg/mL versus 36.2±9.1 pg/mL), respectively. CONCLUSION: These findings suggest that Candida antigens trigger a Th2 instead of Th1 cytokine response in patients with CMC. For better understanding, further studies require on a larger number of patients into the future.