Antimicrobial and anti-biofilm properties of oleuropein against Escherichia coli and fluconazole-resistant isolates of Candida albicans and Candida glabrata.

BACKGROUND: Side effects associated with antimicrobial drugs, as well as their high cost, have prompted a search for low-cost herbal medicinal substances with fewer side effects. These substances can be used as supplements to medicine or to strengthen their effects. The current study investigated the effect of oleuropein on the inhibition of fungal and bacterial biofilm in-vitro and at the molecular level. MATERIALS AND METHODS: In this experimental study, antimicrobial properties were evaluated using microbroth dilution method. The effect of oleuropein on the formation and eradication of biofilm was assessed on 96-well flat bottom microtiter plates and their effects were observed through scanning electron microscopy (SEM). Its effect on key genes (Hwp1, Als3, Epa1, Epa6, LuxS, Pfs) involved in biofilm formation was investigated using the quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) method. RESULTS: The minimum inhibitory concentration (MIC) and minimum fungicidal/bactericidal concentration (MFC/MBC) for oleuropein were found to be 65 mg/ml and 130 mg/ml, respectively. Oleuropein significantly inhibited biofilm formation at MIC/2 (32.5 mg/ml), MIC/4 (16.25 mg/ml), MIC/8 (8.125 mg/ml) and MIC/16 (4.062 mg/ml) (p < 0.0001). The anti-biofilm effect of oleuropein was confirmed by SEM. RT-qPCR indicated significant down regulation of expression genes involved in biofilm formation in Candida albicans (Hwp1, Als3) and Candida glabrata (Epa1, Epa6) as well as Escherichia coli (LuxS, Pfs) genes after culture with a MIC/2 of oleuropein (p < 0.0001). CONCLUSIONS: The results indicate that oleuropein has antifungal and antibacterial properties that enable it to inhibit or destroy the formation of fungal and bacterial biofilm.

Antifungal susceptibility profile of yeasts isolated from the oral cavity of cats.

BACKGROUND: Microorganisms living in the oral cavity play an important role in health and disease of the host. Cats are susceptible to oral infections, and it is documented that fungi in the oral cavity could impact these infections. Antifungal resistance has been increasing in recent years. OBJECTIVES: This study was designed to identify yeast isolates from the oral cavity of healthy cats and to evaluate their antifungal susceptibility pattern. METHODS: Oral specimens were collected from 60 cats and cultured at 37°C for 10 days. Yeasts were isolated and identified. Their antifungal susceptibility pattern was determined according to CLSI M44-A. RESULTS: Three yeast genera were isolated, including Candida spp (55.5%), Rhodotorula spp (33.3%) and Hanseniaspora spp (11.1%). Antifungal susceptibility profiling showed that, apart from a dose-dependent effect of itraconazole, Hanseniaspora spp was susceptible to all seven drugs studied. The Candida species were susceptible to all drugs except ketoconazole (sensitivity 80%) and caspofungin (sensitivity 40%). In R. glutinis and R. minuta, 100% sensitivity was observed for amphotericin B, posaconazole, ketoconazole and voriconazole. CONCLUSIONS: The results suggest that, in comparison with humans and other animals, cats have a different oral mycoflora in terms of species, number and diversity. However, these isolates have similar susceptibility patterns to those seen in isolates from other animals and humans. More studies should be done to further characterize the oral mycobiota of cats and its role in oral infections.

Targeted chitosan nanoparticles embedded into graphene oxide functionalized with caffeic acid as a potential drug delivery system: New insight into cancer therapy.

As a novel drug delivery technology, chitosan (CHI) nanoparticles are encapsulated in graphene oxide (GO) with caffeic acid (CA). The nanocarrier technique combines targeted drug delivery with molecular imaging to provide new cancer insights. Attachment of CA, an anticancer agent for controlled drug release, to functionalized graphene oxide (GON) utilizing 3-aminopropyltriethoxysilane (APTES) was followed by encapsulation of GO with folic acid (FA) attached CHI to produce this novel system. FT-IR was used to characterize and confirm the chemical production process. Brunau-Emmet-Teller (BET) analysis was used to validate multi-holes and nanometric dimensions (1-100 nm) and assess their drug administration use. Release and loading tests showed a pH dependence and implied CA hydrogen-bonding in GON. CA encapsulation and loading percentages are 86 % and 67 %, respectively. The acidic environment (pH 5.3) of tumor cells may produce a larger release of CA, and the release rate of CA maintains a constant trend, indicating the drug is released for more than a week (because the release rate has not reached zero). The proposed method provides a potential candidate for a novel drug delivery system in cancer therapy. The resulting nanohybrid system is a new way to combine biodegradable materials, that can be used in biomedical applications.

Comparative trachea transcriptome analysis in SPF broiler chickens infected with avian infectious bronchitis and avian influenza viruses.

Infectious bronchitis virus (IBV) and avian influenza virus (AIV) are two major respiratory infections in chickens. The coinfection of these viruses can cause significant financial losses and severe complications in the poultry industry across the world. To examine transcriptome profile changes during the early stages of infection, differential transcriptional profiles in tracheal tissue of three infected groups (i.e., IBV, AIV, and coinfected) were compared with the control group. Specific-pathogen-free chickens were challenged with Iranian variant-2-like IBV (IS/1494), UT-Barin isolates of H9N2 (A/chicken/Mashhad/UT-Barin/2017), and IBV-AIV coinfection; then, RNA was extracted from tracheal tissue. The Illumina RNA-sequencing (RNA-seq) technique was employed to investigate changes in the Transcriptome. Up- and downregulated differentially expressed genes (DEGs) were detected in the trachea transcriptome of all groups. The Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology databases were examined to identify possible relationships between DEGs. In the experimental groups, upregulated genes were higher compared to downregulated genes. A more severe immune response was observed in the coinfected group; further, cytokine-cytokine receptor interaction, RIG-I-like receptor signaling, Toll-like receptor signaling, NOD-like receptor signaling, Janus kinase/signal transducer, and activator of transcription, and apoptotic pathways were important upregulated genes in this group. The findings of this paper may give a better understanding of transcriptome changes in the trachea during the early stages of infection with these viruses.

Effects of Bunium persicum essential oil on the reduction of spore germination, growth, and expression of FUM1 and FUM14 genes in Fusarium verticillioides isolates.

BACKGROUND AND PURPOSE: Black Cumin of Kerman (Bunium persicum) is an Iranian plant that is commonly used as an antispasmodic, carminative, and antimicrobial substance. The present study aimed to assess different components of the essence of B. persicum and its effect on antifungal activity, spore germination inhibition, and expressions of FUM1 and FUM14 genes in Fusarium verticillioides strains. MATERIALS AND METHODS: The essence was extracted by hydrodistillation and analyzed through gas chromatography-mass spectroscopy. A broth microdilution method was used for the determination of the minimum inhibitory concentration (MIC). In addition, the expression of FUM1 and FUM14 genes of toxigenic F. verticillioides was assessed by using the real-time polymerase chain reaction (RT-PCR) technique. RESULTS: Based on the findings, most of the essence consisted of γ-terpinene (15.56%), propanal, and 2-methyl-3-phenyl (14.18%). The oil showed a good antifungal activity (mean MIC value: 2556.8 µg/ml) as well as the inhibition of spore germination and mycelial growth (P<0.05). The RT-PCR demonstrated that the expression levels of FUM1 and FUM14 of B. persicum-treated F. verticillioides were 0.43 and 0.53 folds lower than the control samples, respectively. CONCLUSION: These findings revealed that the essential oil of B. persicum has different components responsible for the inhibition of mycelial growth and spore germination of F. verticillioides as well as reduction of expressions of FUM1 and FUM14 genes involving fumonisin production.

Effect of microencapsulation on Saccharomyces cerevisiae var. boulardii viability in the gastrointestinal tract and level of some blood biochemical factors in wistar rats.

BACKGROUND AND OBJECTIVES: Probiotics are live microorganisms that, when administered in an adequate amount, confer a health benefit on the host through the gut. Saccharomyces cerevisiae is a widespread yeast found in nature. This microorganism has been used as a probiotic agent in recent years. In this study, the effect of microencapsulation on survival rate of S. cerevisiae var. boulardii in the simulated gastrointestinal tract medium and the impact of microencapsulated S. cerevisiae var. boulardii on some serum biochemical factors in a rat model was evaluated. MATERIALS AND METHODS: 30 male wistar rats were divided into three groups (control, rats receiving microencapsulated S. cerevisiae var. boulardii, and rats receiving S. cerevisiae var. boulardii alone). The probiotic was gavaged at a dosage of 2 gr/kg BW for 8 weeks. Blood was collected from rats at the end of the treatment period and biochemical factors were measured using Mancompany kits. RESULTS: The results showed a significant increase in viability of microencapsulated S. cerevisiae var. boulardii in comparison with free S. cerevisiae var. boulardii (p<0.05). Weight of rats in probiotic treated groups was significantly higher in comparison with the control group (p<0.05). Moreover, probiotic treatment reduced mean levels of triglycerides, cholesterol, free blood sugar and liver enzymes in rats. CONCLUSION: Microencapsulation could increase the survival rate of yeast probiotics in the gastrointestinal tract; however, more studies are needed for better understanding of the exact effect of microencapsulation on probiotics' function.

Synergistic anticandidal activity of menthol in combination with itraconazole and nystatin against clinical Candida glabrata and Candida krusei isolates.

BACKGROUND: Candida glabrata (C. glabrata) and C. krusei are now emerging as serious hospital acquired infections in immunocompromised patients. Menthol, a terpenic compound, has been reported to have antifungal activity. OBJECTIVES: The aim of this study was to investigate the effect of menthol in combination with itraconazole or nystatin against C. glabrata and C. krusei isolates. METHODS: The effects of menthol along with itraconazole and nystatin, were evaluated by the Clinical Laboratory Standards Institute (CLSI) M44-A and CLSI M27-A3 methods. The fractional inhibitory concentration index (FICI) was determined for menthol plus itraconazole and nystatin combinations using the checkerboard method. RESULTS: The mean of minimum inhibitory concentration (MIC) values of menthol, nystatin and itraconazole were 53.2, 2.30 and 1.50 µg/ml for C. glabrata isolates and 121, 1.08 and 0.38 µg/ml for C. krusei isolates, respectively. Menthol in combination with itraconazole or nystatin exhibited the synergistic effects against all species of Candida tested. FICI values for menthol plus itraconazole and nystatin combinations ranged from 0.250 to 0.561 and 0.139 to 0.623 for C. glabrata isolates, and 0.182 to 0.750 and 0.188 to 0.760 for C. krusei, respectively. CONCLUSIONS: These results support the potential use of menthol as an anticandidal agent, and it can be used complementarily with other conventional antifungal agents.

In vitro lymphoproliferative response and cytokine production in mice with experimental disseminated candidiasis.

OBJECTIVES: Systemic candidiasis is an infection of Candida albicans (C. albicans) causing disseminated disease and sepsis, invariably when host defenses are compromised. We investigated the histopathological changes as well as the lymphoproliferative responses and cytokine production of splenic cells after stimulation with Concanavalin A (Con A) and Pokeweed mitogen (PWM) in mice with disseminated candidiasis. MATERIALS AND METHODS: Lymphoproliferative responses were stimulated in vitro with Con A (1 µg/ml) and PWM (1 µg/ml) mitogens in Roswell Park Memorial Institute (RPMI) 1640 media, and the production of interferon (IFN)-γ and interleukin-4 (IL-4) in the supernatants was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The results revealed that C. albicans organisms multiplied to a greater extent in the kidneys than in the liver and spleen of infected mice. The most predominant forms of C. albicans in different parts of the kidneys were yeast mixed with hyphal forms. Infected mice had a significantly increased proliferative response when splenocytes were stimulated with PWM (2.0±0.16) and Con A (1.9±0.19) (P<0.05). PWM and Con A-stimulated production of IFN-γ significantly tended to be higher in infected mice (PWM: 68.4±14.0 pg/ml; Con A: 53.7±17.3 pg/ml) when compared to controls (P<0.05). Stimulation with PWM and Con A showed no differences in IL-4 production between infected mice and controls. CONCLUSION: These findings demonstrated a significant increase in both cell proliferation and IFN-γ secretion in supernatants of PWM and Con A- stimulated splenocyte cultures obtained from mice with disseminated candidiasis.

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.

Validation of Simplified Tools for Assessment of Sodium Intake in Iranian Population: Rationale, Design and Initial Findings.

BACKGROUND: The 24-hour urine sodium excretion is considered the gold standard method to estimate salt intake. However, since this method is not easy to perform, this study developed two instruments, including a semi-quantitative food frequency questionnaire (FFQ) and one spot urine sodium excretion, to assess sodium intake. These two methods were then compared with 24-h urine sodium excretion and twelve 24-h recalls during a year. METHODS: This study was performed on 219 healthy subjects aged ≥ 6 years in 2014-2015. The FFQ was completed twice, at baseline and one year thereafter, to examine the reproducibility of the FFQ. The validity of three spot urine sodium excretions in the morning, afternoon, and evening and FFQ for the assessment of sodium intake were compared against the 24-h urine sodium excretion method. Moreover, the validity of FFQ was examined against 24-h dietary recalls for the assessment of total sodium consumption and contribution of food groups to sodium intake. The content validity of the FFQ was estimated by an expert panel including 10 nutritionists. RESULTS: Based on their nutrients, the final food items were categorized into 11 groups including: 1) dairy products, 2) fruits, 3) vegetables, 4) meat and egg, 5) grains and legumes, 6) mixed dishes, prepared foods, and restaurant foods, 8) nuts and seeds, 8) oils and fats, 9) sauces and desserts, 10) drinks, and 11) others. CONCLUSIONS: Spot urine and a specific FFQ comprising 136 items were used to develop a method for the assessment of sodium intake and contribution of foods to its intake among the Iranian population. This method can be used in large-scale population studies at the national level.

Chitin-Induced Airway Epithelial Cell Innate Immune Responses Are Inhibited by Carvacrol/Thymol.

Chitin is produced in large amounts by fungi, insects, and other organisms and has been implicated in the pathogenesis of asthma. Airway epithelial cells are in direct contact with environmental particles and serve as the first line of defense against inhaled allergens and pathogens. The potential contributions of airway epithelial cells to chitin-induced asthma remain poorly understood. We hypothesized that chitin directly stimulates airway epithelial cells to release cytokines that promote type 2 immune responses and to induce expression of molecules which are important in innate immune responses. We found that chitin exposure rapidly induced the expression of three key type 2-promoting cytokines, IL-25, IL-33 and TSLP, in BEAS-2B transformed human bronchial epithelial cells and in A549 and H292 lung carcinoma cells. Chitin also induced the expression of the key pattern recognition receptors TLR2 and TLR4. Chitin induced the expression of miR-155, miR-146a and miR-21, each of which is known to up-regulate the expression of pro-inflammatory cytokines. Also the expression of SOCS1 and SHIP1 which are known targets of miR-155 was repressed by chitin treatment. The monoterpene phenol carvacrol (Car) and its isomer thymol (Thy) are found in herbal essential oils and have been shown to inhibit allergic inflammation in asthma models. We found that Car/Thy inhibited the effects of chitin on type 2-promoting cytokine release and on the expression of TLRs, SOCS1, SHIP1, and miRNAs. Car/Thy could also efficiently reduce the protein levels of TLR4, inhibit the increase in TLR2 protein levels in chitin plus Car/Thy-treated cells and increase the protein levels of SHIP1 and SOCS1, which are negative regulators of TLR-mediated inflammatory responses. We conclude that direct effects of chitin on airway epithelial cells are likely to contribute to allergic airway diseases like asthma, and that Car/Thy directly inhibits epithelial cell pro-inflammatory responses to chitin.

Differential Gene Expression of Heat Shock Protein 90 (Hsp90) of Candida albicans obtained from Malaysian and Iranian Patients.

BACKGROUND: Candida albicans (C. albicans) has several virulence factors, in particular heat shock protein 90 (Hsp90), which is expressed by Hsp90 gene. The purposes of this study were to assess the expression of Hsp90 gene in clinical and control isolates of C. albicans obtained from different geographical regions (Malaysia and Iran), different temperatures (25°C, 37°C and 42°C) and mice with candidiasis. METHODS: C. albicans isolates were cultured onto sabouraud dextrose agar (SDA). The assessment of the expression of Hsp90 gene was performed using real time-polymerase chain reaction (RT-PCR). RESULTS: The results showed a significant increase in the expression of C. albicans Hsp90 gene under high thermal shock (42°C) when compared to other temperatures tested (P-value = 0.001). The mean differences in the expression of Hsp90 gene at 37°C were 0.20 (95% confidence interval (CI) 0.13-0.29) between Malaysian and Iranian controls (P-value = 0.040) and 0.47 (95% CI 0.27-0.60) between Malaysian and Iranian patients (P-value = 0.040). CONCLUSION: The results demonstrated that the expression of C. albicans Hsp90 gene varied between Malaysian and Iranian subjects, representing the efficacy of geographical and thermal conditions on virulence gene expression.

In vitro evaluation of antifungal susceptibility and keratinase, elastase, lipase and DNase activities of different dermatophyte species isolated from clinical specimens in Iran.

The aims of this study were to evaluate the enzymatic activity of various dermatophyte species and their antifungal susceptibility profiles. A total of 60 dermatophyte isolates, including Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis and Microsporum gypseum, were examined. Fungal isolates were analysed for the production of keratinase, lipase, elastase and deoxyribonuclease (DNase). A broth microdilution method was performed on the basis of M38-A2 Clinical and Laboratory Standards Institute (CLSI) guidelines. T. mentagrophytes, M. canis and M. gypseum isolates were capable of producing keratinase, lipase, elastase and DNase, while T. rubrum isolates were elastase negative. The highest mean diameter of the clear zone around the colonies (PZ) was associated with keratinase (PZ: 4.56 ± 1.29 mm), followed by lipase (PZ: 1.53 ± 0.90 mm), DNase (PZ: 0.65 ± 0.54 mm) and elastase (PZ: 0.22 ± 0.27 mm) (P < 0.05). The mean minimum inhibitory concentration 90 (MIC90 ) of all strains were as follows: itraconazole (MIC90 : 0.28 ± 0.31 µg ml-1 ), ketoconazole (MIC90 : 0.48 ± 0.51 µg ml-1 ), griseofulvin (MIC90 : 0.86 ± 1.00 µg ml-1 ) and fluconazole (MIC90 : 18.57 ± 20.10 µg ml-1 ). Dermatophyte isolates had higher keratinolytic activity than other enzymes. Itraconazole was the most effective antifungal drug and fluconazole had the poorest activity.

A comparative study of the timecourse of the expression of the thermo­inducible HSP70 gene in clinical and environmental isolates of Aspergillus fumigatus.

The internal environment within animals or humans provides different conditions to invading saprophytic fungal pathogens, requiring the differential regulation of genes in comparison to environmental conditions. Understanding the mechanisms by which pathogens regulate genes within the host may be key in determining pathogen behavior within the host and may additionally facilitate further investigation into novel therapeutic agents. The heat shock protein (HSP)70 gene and its associated proteins have been frequently reported to be among the most highly expressed and dominant proteins present within various locations at physiological temperatures. The present study examined relative gene expression levels of the HSP70 gene in Aspergillus fumigatus isolates from both clinical and environmental origins, at a range of temperature points (20, 30, 37 and 42˚C) over five days, using reverse transcription­quantitative polymerase chain reaction, comparing with a standard A. fumigatus strain incubated at 25˚C. The results indicated a differential gene expression pattern for the environmental and clinical isolates. During the five days, the HSP70 expression levels in the clinical samples were higher than in the environmental samples. However, the difference in the expression levels between the two groups at 42˚C was reduced. The mean HSP70 expression level over the five incubation days demonstrated a gradual and continual increasing trend by temperature elevation in both groups at 30, 37 and 42˚C, however, at 20˚C both groups demonstrated reduced expression. The temperature shift from 20 to 42˚C resulted in HSP70 induction and up to a 10­ and 8.6­fold change in HSP70 expression levels on the fifth day of incubation in the clinical and environmental groups, respectively. In conclusion, incubation at 37 and 42˚C resulted in the highest expression levels in both experimental groups, with these temperature points important for the induction of HSP70 expression in A. fumigatus.

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.

Study on mycoflora of poultry feed ingredients and finished feed in Iran.

BACKGROUND AND OBJECTIVES: Unhygienic poultry feedstuffs can lead to nutrient losses and detrimental effect on poultry production and public health. In the present study, mycobiota and colony-forming units per gram in ingredients and finish poultry feed was evaluated with special reference to potentially mycotoxigenic fungi. MATERIALS AND METHODS: Eighty five samples of corn, soybean meal and poultry finished feed were collected from nine poultry feed factories located in three provinces i.e. Tehran, Alborz and Qom in Iran from October 2014 to January 2015. Samples were cultured on Sabouraud dextrose agar (SDA), Aspergillus flavus and parasiticus agar (AFPA) and dichloran rosebengal chloramphenicol agar (DRBC) and incubated at 28 °C for 7-10 days. Purified fungal colonies were identified by a combination of macro- and microscopic morphological criteria. For determining the rate of fungal contamination, samples were cultured on SDA and colony forming units (CFUs) were calculated. RESULTS: A total of 384 fungal isolates belonging to 7 genera of filamentous fungi and yeasts were obtained from corn (124 isolates), soybean meal (92 isolates), and feed before (72 isolates), and after pelleting (96 isolates). The most prominent fungal isolate in corn, soybean meal and feed before pelleting (feed as mash form) was Fusarium but in feed after pelleting was Aspergillus. Among 5 Aspergillus species isolated, potentially aflatoxigenic A. flavus isolates was predominant in corn (46.6%), soybean meal (72.7%) and poultry finished feed (75%). CFUs results indicated that 9/22 corn samples (40.9%), none of 22 soybean meal samples, 19/41 finished feed (46.3%) were contaminated higher than the standard limit. CONCLUSIONS: Our results indicated that corn, soybean meal and finished feed of poultry feed mill are contaminated with various fungal genera by different levels sometimes higher that the standard limits. Contamination with potentially mycotoxigenic fungi especially Aspergillus species may be considered as a human public health hazard.

Factors in Etiology and Predisposition of Adult Tinea Capitis and Review of Published Literature.

BACKGROUND: Tinea capitis is a common fungal infection in children but is less frequently encountered in adults, especially in immunocompromised individuals. OBJECTIVES: To determine the incidence of tinea capitis in adults, the predisposing factors and causative species. METHODS: A retrospective study was conducted over a period of 5 years, from 2010 to 2015, on cases of tinea capitis diagnosed in the Department of Dermatology and Mycology Research Center in Tehran, Iran. The information was collected from the patients including age, gender, location of the lesions, results of direct examination and culture, cause of immunosuppression and the prescribed treatment. RESULTS: Twenty-five (20.6 %) patients (10 men and 15 women) with a mean age of 45.28 years were affected by tinea capitis among a total number of 121 positive cases. Most of these adults (80 %) had a grade of immunodeficiency due to the underlying syndromes or diseases, and the rest were immunocompetent. Trichophyton species were isolated from 84 % of these adult patients, indicating Trichophyton violaceum (T. violaceum) as the most common fungal agent. Treatment with oral terbinafine or itraconazole was successful in all these cases. CONCLUSIONS: The results showed that most cases affecting the adult population were caused by species of the genus Trichophyton. T. violaceum was the most common dermatophyte of adult patients. Thus, it is important to consider tinea capitis as a differential diagnosis in immunocompromised adults, even though it is considered to be rare in adults.

Efficacy of Cuminum cyminum essential oil on FUM1 gene expression of fumonisin-producing Fusarium verticillioides strains.

OBJECTIVES: The purpose of this study was to evaluate the effect of Cuminum cyminum (C. cyminum) essential oil on the growth and FUM1 gene expression of fumonisin-producing Fusarium verticillioides (F. verticillioides) strains isolated from maize. MATERIALS AND METHODS: All fungal strains were cultured on potato dextrose agar (PDA) slopes at 30°C for 7 days. The antifungal activity was evaluated by broth microdilution assay. One set of primers was F. verticillioides species specific, which selectively amplified the intergenic space region of rDNA. The other set of primers was specific to FUM1 gene region of fumonisin-producing F. verticillioides. FUM1 transcript levels were quantified using a reverse transcription-polymerase chain reaction (RT-PCR) protocol. RESULTS: Minimum inhibitory concentration (MIC) values of C. cyminum oil against F. verticillioides strains varied from 0.195 to 0.781 µl.ml(-1) (mean MIC value: 0.461 µl.ml(-1)) indicating 54.5% of the fungal strains inhibited at 0.390 µl.ml(-1). PCR analysis of FUM1 gene expression revealed that DNA fragment of 183 bp was amplified in all the isolates of F. verticillioides before treatment with C. cyminum essential oil. Based on RT-PCR analyses, reduction in the expression of fumonisin biosynthetic genes was significant only for FUM1 gene (p<0.05), while no effect was observed on ITS gene. CONCLUSIONS: This study showed that all F. verticillioides isolates were susceptible to C. cyminum essential oil, indicating a significant reduction in the growth of fungal isolates. In addition, this oil completely inhibited the expression of FUM1 gene in concentrations dose-dependently.

Microencapsulation of Saccharomyces cerevisiae and its evaluation to protect in simulated gastric conditions.

BACKGROUND AND OBJECTIVES: Probiotic yeasts are used in production of functional foods and pharmaceutical products. They play an important role in promoting and maintaining human health. Until now, little work has been published on improving the survival of Saccharomyces in stimulated gastrointestinal condition. MATERIAL AND METHODS: In this study the exposure of the yeast in the capsulate and free forms to artificial gastrointestinal conditions was assessed and the number of viable Saccharomyces cerevisiae cells during 0 to 120 mines in these conditions was evaluated by a pour plate method using sabouraud dextrose agar. RESULTS: Results showed the shape of the beads was generally spherical, sometimes elliptical with a mean diameter of about 50-90 µm. Also count of viable probiotic cells obtained for all the microcapsules were above the recommended levels for a probiotic food. Also decrease of approximately 4 logs was noted in the number of free cells after 2 h of incubation at pH 2 and 8, when compared to decreases of about 2 logs in the all microencapsulated S. cerevisiae under similar conditions. CONCLUSION: It is concluded that microencapsulation process was significantly able to increase the survival rate of Saccharomyces in a simulated gastrointestinal condition (p<0.05)..

Sequences type analysis of Candida albicans isolates from Iranian human immunodeficiency virus infected patients with oral candidiasis.

The growing number of immunocompromised individuals has increased the incidence of infections caused by Candida species during the recent decades. Typing of C. albicans on the basis of DNA sequences at multiple loci has greatly advanced our knowledge about the epidemiology and phylogeny of candidiasis. The aim of this study was to evaluate the diversity, and genetic relationships among C. albicans isolates obtained from HIV patients in Iran. using multilocus sequence typing (MLST) method. We analyzed 25 C. albicans isolates obtained from HIV positive patients referred to Iranian Research Center for HIV/AIDS. After diagnostic test and DNA extraction C. albicans isolates were typed using the original MLST scheme explained previously include of six loci: ACC1, VPS13, GLN4, ADP1, RPN2, and SYA1. Fifty one (2.17%) nucleotide sites were found to be polymorphic; all were found to be heterozygous in at least one isolate. For the 25 clinical isolates, 22 diploid sequence types were defined by the genotypes identified from the six loci. The MLST data suggest a relatively high level of divergence in the population structure of C. albicans isolated from HIV infected patients. These findings indicate that in these patients there is a favorable context for the growth of potential pathogenic C. albicans. We found no association between fluconazole resistance, highly active antiretroviral therapy (HAART) receiving and either sequence type or group.