Combined effects of fungal β-glucan and Zataria multiflora essential oil on phagocytosis in Balb/C mice

Introduction: Natural substances have been used since ancient times for treatment of a range of diseases and have represented stimulatory effects on the function of innate immunity. The purposes of this study were to prepare β-glucan from S. cerevisiae and to assess the efficacy of purified β-glucan, Zataria multiflora (Z. multiflora) essential oil and their complex on phagocytosis in Balb/c mice.Methods: β-glucan was purified during three stages including alkaline-acid treatment (S1), DEAE sephacel chromatography (S2) and con-A sepharose chromatography (S3). Z. multiflora essential oil was extracted by water-distillation using Clevenger-type apparatus. The chemical composition of Z. multiflora essential oil was analyzed by a GC/MS system. β-glucan (15 mg/kg), Z. multiflora essential oil (100 mg/kg) and their complex (the same doses) were injected into Balb/c mice intraperitoneally (IP). The blood samples were collected at days 4 and 7 after injection and phagocytic activity was assayed by chemiluminescence method.Results: The results showed that the ratios of mannan to β-glucan were 70.3 to 29.7 for S1, 71.9 to 28.1 for S2 and zero to 100 for S3 (purified β-glucan). The major components of Z. multiflora were carvacrol (61) and thymol (25). Phagocytosis index means exhibited significant increases at day 4 (246, 165 and 367) and day 7 (242, 235 and 309) in mice treated with purified β-glucan, Z. multiflora essence, and their complex when compared to control mice, respectively (P < 0.05).Conclusion: The results suggest that the complex of β-glucan and Z. multiflora oil might be used to immunize individuals as prophylactic and/or therapeutic adjuvant in immunocompromised subjects

Zataria multiflora Boiss.: A review study on chemical composition, anti-fungal and anti-mycotoxin activities, and ultrastructural changes

Introduction: Zataria multiflora is a valuable medicinal plant from Lamiaceae family with various pharmacological and therapeutic properties. In this article we reviewed the various aspects of Z. multiflora properties including botanical characteristics, chemical composition, anti-fungal and anti-mycotoxin activity and fungal ultrastructural changes.Methods: Google Scholar, PubMed, EBSCO, Directory of open access journals (DOAJ), EMBASE, and Web of Science were searched using the keywords Z. multiflora and pathogenic and toxigenic fungi.Results: The essential oil (EO) of Z. multiflora is frequently used in pharmaceutical industries. Thymol and carvacrol are the most important active components of Z. multiflora that are used for the treatment of a wide variety of diseases such as candidiasis and dermatophytosis. Aflatoxin production inhibitory effect of Z. multiflora EO was at the transcription level and this herb may cause reduction in aflatoxin biosynthesis. Ultrastructural changes showed that the main sites of action of EO were the plasma membrane and cell wall of fungi.Conclusion: Zataria multiflora has the potential to be considered as a new natural drug for the treatment of some fungal infections. Morphological and structural changes may be one of the mechanisms involved in growth inhibition of the fungi and reducing aflatoxin production by Z. multiflora EO

Inhibitory effect of plant essential oils on Malassezia strains from Iranian dermatitis patients

Introduction: The genus Malassezia is an important skin resident of human. The present study aimed to analyze in vitro activity of the essential oils of Lavandula stoechas, Cuminum cyminum and Artemisia sieberi against clinical strains of Malassezia species. Methods: A total of 47 Malassezia strains, including Malassezia furfur, Malassezia globosa and Malassezia obtuse, were used in this study. A disk diffusion technique was selected for testing the susceptibility of Malassezia strains to the essential oils. Results: All the essential oils showed in vitro activity against Malassezia strains, with M. furfur and M. obtusa being the highest and lowest susceptible of the strains, respectively. The highest antifungal activity was associated with the essential oil of C. cyminum (mean ± SD: 50.0 ± 0.0 mm), followed by L. stoechas (mean ± SD: 46.8 ± 3.1 mm) and A. sieberi (mean ± SD: 36.9 ± 5.7 mm). The inhibition zone ranges were 12.5 to 15.6 mm (mean ± SD: 14.4 ± 1.6 mm) for ketoconazole and 11.6 to 13.3 mm (mean ± SD: 12.4 ± 0.9 mm) for fluconazole. Although all the antifungal drugs were found to possess good antifungal activities against Malassezia strains, their effects were lower than the activities shown by the essential oils tested (P<0.05). Conclusion: These results indicated that the essential oils tested, especially the one from C. cyminum, inhibited the growth of clinical strains of Malassezia, implying its potential use in the treatment of Malassezia infections. This indicates that this plant may be useful in preparation of new drugs

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 82 kDa. The least average concentration ratios of Hsp90 were 0.350 and 0.240 mg/g for Malaysian and Iranian isolates at 25 °C, respectively, while the highest average concentration ratios of Hsp90 were 3.05 and 2.600 mg/g for Malaysian and Iranian isolates at 42 °C, respectively. There were differences in the ratio amount of Hsp90 between Malaysian isolates (1.01 ± 0.07 mg/g) and mice kidneys (1.23 ± 0.28 mg/g) as well as between Iranian isolates (0.70 ± 0.19 mg/g) and mice kidneys (1.00 ± 0.28 mg/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

In vitro antifungal effect of herbal mixture (Nigella sativa, Foeniculum vulgare and Camellia sinensis) against Candida species isolated from denture wearers

Introduction: Due to antimicrobial and dental plaque control activities, herbal mouthwashes lead to an improvement in oral health. Although chemical mouthwashes have demonstrated the greatest antimicrobial and anti-inflammatory effects, their usage has been limited because of their numerous side effects. The aim of this study was to assess the antifungal activity of herbal mixtures containing Nigella sativa, Foeniculum vulgare and Camellia sinensis against oral isolates of Candida from denture wearers.Methods: We selected 93 individuals wearing complete denture prosthesis. Samples were collected from oral mucosa and dentures and cultured onto Sabouraud dextrose agar (SDA). The antifungal activities of N. sativa, F. vulgare and C. sinensis and their mixtures (no. 1, 2 and 3) against oral isolates of Candida were determined using punch-hole test.Results: The oral cavities of all denture wearers were colonized with yeasts. Among the Candida species, Candida albicans was the most frequently recovered species (45; 48.4), followed by C. tropicalis (14; 15), C. krusei (9; 9.7), C. glabrata (6; 6.5), C. dubliniensis (4; 4.3) and Candida spp. (15; 16.1). Among the tested plants, N. sativa (mean value: 12.3 mm) and F. vulgare (mean value: 7.9 mm) showed positive results against all Candida isolates. The results exhibited that all herbal mixtures were active against various tested Candida isolates, ranging from 7.8 to 15 mm, 7.6 to 15.5 mm and 7 to 15 mm inhibition zones for herbal mixtures no. 1, 2 and 3, respectively.Conclusion: The results indicated that C. albicans was the most prevalent Candida species. N. sativa and F. vulgare were good antifungal agents against oral species of Candida isolated from individuals wearing complete dentures, hence, there is a possible usefulness as therapeutic agents

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

Fungicidal effect of Origanum vulgare essential oil against Candida glabrata and its cytotoxicity against macrophages

Introduction: Candida glabrata is a yeast fungus regularly isolated from patients with impaired immunity who receive a routine antifungal therapy. Drug-resistant strains of C. glabrata have been emerged in recent years. The aim of this study was to examine the therapeutic efficacy Origanum vulgare essential oil (OVEO) against drug-resistant strains of C. glabrata and its cytotoxic effect on macrophages.Methods: Specimens were collected from mucosal surfaces of the oral cavity of medically approved oropharyngeal candidiasis (OPC) in HIV-positive patients and volunteered healthy individuals using sterile swabs or mouthwashes. In vitro antifungal susceptibility testing was done using microdilution and disc diffusion methods. Chemical composition of OVEO was determined using gas chromatography mass spectrometry. The cytotoxic effect of essential oil on macrophages was examined using tetrazolium dye (MTT).Results: Minimum inhibitory concentration (MIC) range of OVEO in healthy individuals and OPC patients was 150-200 and 150-250 &mu;g/mL, respectively. OVEO efficiently inhibited growth of resistant isolates. In isolates obtained from HIV patients, both MIC50 and MIC90 of OVEO were 200 &mu;g/mL while in healthy individuals were 150 and 200 &mu;g/mL, respectively. Moreover, OVEO induced significant reduction in proliferation of murine RAW264.7 and peritoneal macrophages in concentrations higher than 100 and 300 &mu;g/mL, respectively. Main constituents of OVEO were thymol (27.3), &gamma;-terpinene (20.7) and carvacrol (16.1).Conclusion: OVEO could be used as a fungicidal agent against fungal infections caused by azole-resistant C. glabrata. A combination therapy along with standard antifungals is suggested to avoid its cytotoxic effects.</p

FREQUENCY OF YEASTS AND FILAMENTOUS FUNGI IN THE EXTERNAL EAR CANALS OF CATTLE IN IRAN

  Fungal microbiotas are saprophytic microorganisms that can act as opportunistic pathogens in animals. This study was carried out in order to isolate and identify the ear fungal biota from healthy cattle. The samples were taken using premoistened swabs from the right and/or left external ear canals of 32 healthy cattle and cultured onto Sabouraud glucose agar and modified Dixon's agar media. A total of eight different fungal genera were isolated from 29 (90.6%) of 32 healthy cattle. Both filamentous fungi and yeasts were isolated with the predominance of Aspergillus spp. (35.6%), Candida spp. (18.9%) and Malassezia spp. (16.8%). The most frequent Aspergillus spp. were A. fumigatus (16.8%), A. glaucus (14.9%) and A. flavus (4%). Among the fungal isolates, 46.5% and 17.8% colonies were associated with hyaline and dematiaceous fungi, respectively (p = 0.003). The recognized fungi, especially Aspergillus spp. and Candida spp., were colonizes as mycobiota in the external ear canals of healthy cattle, leading to otitis externa under predisposing factors

A review on the inhibitory potential of Nigella sativa against pathogenic and toxigenic fungi

Nigella sativa (N. sativa) grows in various parts of the world, particularly in Iran. It has been traditionally used as a folk remedy to treat a number of diseases. The seeds of this plant contain moisture, proteins, carbohydrates, crude fiber, alkaloids, saponins, ash, fixed oils and essential oil. The major components of the essential oil are thymoquinone, p-cymene, trans-anethole, 2-methyl-5(1-methyl ethyl)-Bicyclo[3.1.0]hex-2-en and γ-terpinene. So far, several pharmacological effects such as anti-oxidant, anti-inflammatory, anti-cancer and anti-microbial have been reported for N. sativa or its active compounds. Thymoquinone, thymohydroquinone and thymol are the most active constituents which have different beneficial properties. The oil, extracts and some of N. sativa active components possessed moderate in vitro and in vivo inhibitory activity against pathogenic yeasts, dermatophytes, non-dermatophytic filamentous fungi and aflatoxin-producing fungi. The main morphological changes of pathogenic and toxigenic fungi treated with N. sativa oil were observed in the cell wall, plasma membrane and membranous organelles, particularly in the nuclei and mitochondria. Although this review represents first step in the search for a new anti-fungal drug, the full potential of N. sativa as a fungitoxic agent has not been exploited and necessitates further investigations

Zataria multiflora Boiss.: A review study on chemical composition, anti-fungal and anti-mycotoxin activities, and ultrastructural changes

Introduction: Zataria multiflora is a valuable medicinal plant from Lamiaceae family with various pharmacological and therapeutic properties. In this article we reviewed the various aspects of Z. multiflora properties including botanical characteristics, chemical composition, anti-fungal and anti-mycotoxin activity and fungal ultrastructural changes. Methods: Google Scholar, PubMed, EBSCO, Directory of open access journals (DOAJ), EMBASE, and Web of Science were searched using the keywords Z. multiflora and pathogenic and toxigenic fungi. Results: The essential oil (EO) of Z. multiflora is frequently used in pharmaceutical industries. Thymol and carvacrol are the most important active components of Z. multiflora that are used for the treatment of a wide variety of diseases such as candidiasis and dermatophytosis. Aflatoxin production inhibitory effect of Z. multiflora EO was at the transcription level and this herb may cause reduction in aflatoxin biosynthesis. Ultrastructural changes showed that the main sites of action of EO were the plasma membrane and cell wall of fungi. Conclusion: Zataria multiflora has the potential to be considered as a new natural drug for the treatment of some fungal infections. Morphological and structural changes may be one of the mechanisms involved in growth inhibition of the fungi and reducing aflatoxin production by Z. multiflora EO