Biological Properties of Vitex agnus-castus Essential Oil (Phytochemical Component, Antioxidant and Antifungal Activity)

Background: Vitex agnus-castus is a deciduous shrub that is native to the Mediterranean region. It has traditionally been used in Iranian medicine. In the current study, Vitex agnus-castus Essential Oil (EO) leaves were analyzed for their chemical component as well as antioxidant and antifungal activity. Objectives: The aim of this study was to determine the biological properties (phytochemical component, and antioxidant and antifungal activity) of Vitex agnus-castus EO from an Iranian origin. Materials and Methods: Chemical composition of the EO was determined by using Gas Chromatography (GC) and Gas Chromatography/Mass Spectrometry (GC-MS). Antioxidant activity of the Vitex agnus-castus EO was examined by the 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) assay, while the total phenolic content was also determined. Antifungal activity (against Candida albicans, Candida tropicalis, Candida parapsilosis, Candida krusei, Candida dubliniensis, Aspergillus flavus, Aspergillus niger, Penicillium species and Alternaria species) was performed by a broth microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) protocols M27-A and M38-A for yeasts and filamentous species. Results: Thirty-two components were identified in Vitex agnus-castus EO. The main compound was alpha-Pinene (19.48%). The total phenolic content of EO was determined as 82.26 ± 5.94 mg Gallic Acid Equivalent (GAE)/g EO. The EO exhibited significant radical scavenging activity with IC50 value of 27.16 μg/mL. The obtained EO showed significant antifungal activity. Aspergillus niger was more susceptible than other fungi (MIC: 0.78 μL/mL). Conclusions: Potent antifungal activity, make this plant an effective replacement treatment for fungal infections or fungal strains that are resistance to synthetic antifungals. Keywords: Diphenyl-2-picrylhydrazyl; Essential oil; Phenolic Component; Vitex agnus-castus; Gas Chromatography-Mass Spectrometr

Emergence of Terbinafine Resistant Trichophyton mentagrophytes in Iran, Harboring Mutations in the Squalene Epoxidase (SQLE) Gene

Introduction: Trichophyton mentagrophytes and T. interdigitale are important causative agents of superficial mycoses, demonstrating emergent antifungal drug resistance. We studied the antifungal susceptibility profiles in Iranian isolates of these two species. Methods: A total of 96 T. interdigitale and 45 T. mentagrophytes isolates were subjected to molecular typing by ribosomal ITS region. Antifungal susceptibility profiles for terbinafine, griseofulvin, clotrimazole, efinaconazole, luliconazole, amorolfine and ciclopirox were obtained by CLSI broth microdilution method. The squalene epoxidase (SQLE) gene was subjected to sequencing for mutations, if any, in isolates exhibiting elevated MICs for terbinafine. Results: Luliconazole and efinaconazole showed the lowest MIC values against T. mentagrophytes and T. interdigitale isolates. There were five isolates with terbinafine MICs >= 32 mu g/mL in our sample. They belonged to T. mentagrophytes type VIII and harbored two alternative SQLE gene sequence variants, leading to Phe397Leu and Ala448Thr or Leu393Ser and Ala448Thr substitutions in the enzyme. All terbinafine resistant strains could be inhibited by luliconazole and efinaconazole. Conclusion: This study documented a step in the global spread of resistance mechanisms in T. mentagrophytes. However, treatment alternatives for resistant isolates were available. Keywords:Trichophyton mentagrophytes; SQLE; terbinafine; antifungal drug resistance; Ira

Inhibitory Activity of Artemisia spicigera Essential Oil Against Fungal Species Isolated From Minced Meat

Abstract Background: Meat is an important source of several nutrients. The capability top of fresh meat to rot, causing the group of studies food science, biological and chemical stability meat consideration. Objectives: This study was conducted to examine the inhibitory effect of Artemisias spicigera essential oil against fungal species isolated from minced meat. Materials and Methods: Two types of media dichloran 18% glycerol (DG18) agar and dichloran rosebengal chloramphenicol (DRBC) agar were selected for the mycological analysis of the minced meat samples. To evaluate the antifungal activity of essential oils, the microdilution broth method based on the CLSI (M27A) guideline was used. Results: Artemisias spicigera essential oil has an inhibitory effect on the growth of fungi found in samples of minced meat. Aspergillus, Penicillium and Cladosporium were the most common genera on both medium types. Average Minimum Inhibitory Concentration 50 = 1.88 μL/mL and MIC90 = 2 μL/mL were reported. The genus of Mucor with MIC = 1.0 μL/mL was the most sensitive and Aspergilus versicolor was the most resistant species to the essential oil with MIC = 4 μL/mL. Conclusions: The results of the present study show a favorable inhibitory effect of Artemisias spicigera essential oil on fungal growth, especially Aspergillus species. According to the results, antifungal components of Artemisias spicigera in different forms are used to prevent fungal pollution. Keywords: Antifungal Agent, Food Contamination, Meat, Artemisi

Evaluation of Antioxidant, Antibacterial, and Antifungal Properties of Satureja hortensis Essential Oil

Background: Satureja is a genus belonging to the aromatic plants of Lamiaceae family. The genus Satureja L. (Lamiaceae) comprises more than 30 species of aromatic herbs and shrubs, widely distributed over the Mediterranean region. The genus is represented by 14 species in Iran of which, eight are endemic. Many species of the genus Satureja are reported to have aromatic and medicinal properties. The leaves, flowers, and stems of this plant are used as herbal tea and in treatment of various ailments in traditional medicine. Objectives: The aim of this study was to investigate antimicrobial and antioxidant activity of essential oil (EO) of Satureja hortensis (Lamiaceae) that grows in Sabalan Mountain (Ardebil province, Iran). Materials and Methods: This EO was tested in vitro against two bacterial species by disk and agar well diffusion methods and against four bacterial species and three Candida strains by broth microdilution method. Total phenol, flavonoid, and free radical scavenging activity of EO were evaluated. Results: Total phenolic content was determined by using Folin-Ciocalteu reagent (32.65 mg/g), which was presented as gallic acid equivalent in 1 g of sample. IC50 of EO and ascorbic acid in DPPH method were respectively 277.9 and 19.34 μg. Minimum inhibitory concentration of the EO of S. hortensis against Salmonella typhimurium, Listeria monocytogenes, Escherichia coli, and Bacillus cereus was respectively 2.5%, 2.5%, 5%, and 2.5%. The inhibition zone of EO in disk and agar well diffusion method showed that inhibitory zone on B. cereus was higher than that on S. typhimurium in both methods and B. cereus was more sensitive to EO. MIC to minimal fungicidal concentration (MFC) ratio of S. hortensis EO against Candida albicans, Candida parapsilosis, and Candida krusei in broth microdilution method were respectively 0.048%:0.048%, 0.024%:0.024%, and 0.012%:0.012%. Conclusions: The data of the study clearly indicated that the EO of S. hortensis has a strong antioxidant, antibacterial, and antifungal activity. Keywords:Essential Oils; Antioxidants; Traditional Medicin

Emergence of Azoles Resistance Candida species in Iranian AIDS defined patients with oropharyngeal candidiasis

Background and Purpose: Oropharyngeal candidiasis (OPC) and antifungal drug resistance are major health concerns in patients with human immunodeficiency virus (HIV). The increased reports of antifungal resistance and expanding drug therapy options prompted the determination of antifungal susceptibility profile. The present study was performed to determine the antifungal susceptibility of Candida species isolated from AIDS patients with OPC in Iran. Materials and Methods: In total, 100 Candida isolates from the oral cavity of patients with OPC (TCD4 < 200 cells/&muL) were obtained and cultured on CHROMagar and Sabouraud’s dextrose agar. All isolates were identified according to the assimilation profile, colony color and other conventional methods. Broth microdilution of antifungal drugs was carried out, according to the methods described in M27-S4 and M44-A guidelines by the Clinical and Laboratory Standards Institute (CLSI). Results: Among 60 Candida albicans (C. albicans) strains, 56.7% were resistant to fluconazole, while 38.3% were resistant to ketoconazole and clotrimazole. The resistance of C. albicans isolates against polyene antifungals including amphotericin B was scarce (1.7%). Based on the results, 52.2% of C. glabrata strains were resistant to fluconazole, while 47.8% and 30.4% of these isolates were resistant to ketoconazole and clotrimazole, respectively. All Candida isolates were susceptible to nystatin and caspofungin. Conclusion: Based on the findings, it can be concluded that screening of resistant Candida isolates by disk diffusion or broth dilution method is essential for the surveillance and prevention of antifungal resistance in patient management. Although nystatin is widely used in clinical practice for HIV patients in Iran, no evidence of enhanced resistance against this agent was found on the other hand, resistance to azole antifungals, particularly fluconazole, increased. Considering the lack of resistance to caspofungin, administration of this agent is suggested for the treatment of OPC in AIDS patients

Study on inhibitory activity of Artemisia Spergicia essential oil against fungal species isolated from minced meat

Objective: meat is an important source of several nutrients. The ability high corruption of fresh meat has caused some studies food science, biological and chemical stability meat consideration. This research was conducted with the aim of Study on inhibitory activity of Artemisia Spergicia essential oil against fungal species isolated from minced meat In Qazvin city. Methods: Two types of media Dichloran 18% Glycerol agar (DG18) and Dichloran Rosebengal Chloramphenicol agar (DRBC) were chosen for the mycological analysis of the minced meat samples as used. To evaluate the antifungal activity of essential oils, microdilution broth method based on CLSI(M27A) guideline was used. Results: Essential oil (Artemisia Spergicia) inhibitory effect on the growth of fungi was found in samples of minced meat. Aspergillus , Penicillium and Cladosporium were the most common genera on both medium types. Average MIC50=1.88 µl/ml and MIC90=2 µl/ml reported.The genus Mucor with MIC=1.0 µl/ml was the most sensitive and Aspergilus vercikaler most resistant to the essential oil with MIC=4 µl/ml. Conclusion: The results show a favorable inhibitory effect of Artemisia Spergicia essential oil on fungi growth, especially Aspergillus species. According to this result, antifungal components of Artemisia Spergicia in different forms be used to prevent fungi Pollution. Key Words: minced meat, fungal infection, Essential oil of Artemisia, Antifungal activit

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

Genotyping of Candida albicans isolated from animals using 25S ribosomal DNA and ALT repeats polymorphism in repetitive sequence

Background and Purpose: Candida albicans is the most prevalent Candida species isolated from animals. Candidiasis can be systemic in animals or may affect a single organ, such as the mouth, urinary tract, and skin. The aim of the present study was to determine the genetic diversity of C. albicans isolated from different animals and investigate the presence of a relationship between host specificity and genetic typing of C. albicans. Materials and Methods: For the purpose of the study, DNA extraction was performed on 27 clinical isolates of C. albicans obtained from animals. Subsequently, they were subjected to 25S ribosomal DNA amplification and ALT repeats in repetitive sequences (RPSs). The minimum inhibitory concentrations of fluconazole, ketoconazole, clotrimazole, nystatin, amphotericin B, and caspofungin were determined using the microdilution method based on the Clinical and Laboratory Standards Institute M27-S4 standard. Results: Out of 27 C. albicans strains, 11, 6, 5, and 5 cases were recognized as genotypes A (40.8%), E (22.2%), B (18.5%), and C (18.5%), respectively, through amplification using AS-I, which revealed 17 different types of C. albicans. By combining the two typing methods, 27 C. albicans strains were finally divided into 22 genotypes. Conclusion: Different genotypes showed genetic diversity among the C. albicans strains isolated from animal sources. The results revealed no special genotype relationship according to the host, anatomical source of isolation, and antifungal susceptibility

Multilocus sequence typing of Candida albicans isolates from Burn Intensive Care Unit (BICU) in Iran

Burn intensive care unit patients are specifically exposed to deep-seated nosocomial infections caused by Candida albicans. Superficial carriage of C. albicans is a potential source of infection and dissemination, and typing methods could be useful to trace the different isolates. Multilocus sequence typing is a powerful genotyping method for pathogenic microorganisms, including Candida albicans. Thirty clinical isolates of C. albicans obtained from 22 patients that admitted to the Burn Intensive Care Unit (BICU) from a burn hospital at Sari, Mazandaran state, Iran were epidemiologically studied by multilocus sequence typing (MLST). Seventy five variable nucleotide sites were found. Sixty two alleles were identified among the seven loci of the C. albicans isolates and one new allele was obtained. Eighteen Diploid Sequence Types (DSTs) were identified and among those 10 were new. These isolates belonged to nine clonal clusters (CCs) while two isolates occurred as singletons. Eleven (36.7%) isolates belonged to CC 124 after eBURST analysis and 13 isolates (43.3%) were assigned to clade 4. Approximately 17% of the 30 isolates belonged to clade 1 (CC 69 and CC766). Isolates from several patients with burns were found to be genetically related. Some patients yielded multiple isolates with identical DSTs suggest colonization or infection caused by cross contamination between patients. Therefore, isolates that show identical or similar allelic profiles are presumed to be identical or closely related and may use to evaluate the genetic relationships between isolates from a specific environment such as BICU

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

Backgrounds and aims: Candida albicans is more prevalent between Candida species which can cause candidal infection. Various studies have shown drug resistance among the C. albicans isolates, so it is now necessary to discover replacement treatments for the Candida infection. The aim of this study is comparison the effects of different Essential oils (EOs) against azoles-resistant and azoles-susceptible isolates. &nbsp;Materials and methods: Twenty fluconazole resistance and twenty susceptible C. albicans, obtained from oral, vaginal and cutaneous tissues of patients with Candidiasis were evaluated. The efficacy and MICs of Zataria multiflora, Geranium herbarum, Lavendula officinalis, Cuminum, cyminum, Allium heamanthoides and Artemisia sieberi EOs against C. albicans, were determined on the basis of a reference method for broth microdilution susceptibility testing of yeasts, suggested by CLSI-M27S4. After the period of inoculation, incubation and subculturation, the MICs compared to the control tubes, were determined. Results: The obtained MICs for Zataria multiflora, Geranium herbarum, Artemisia sieberi and Lavendula officinalis EOs were between 0.1-0.25 &micro;l/ml (mean: 0.18&micro;l/ml), 0.4-1.3&micro;l/ml (mean: 0.83&micro;l/ml) and 0.7-2.25 &micro;l/ml (mean: 1.4&micro;l/ml), 0.6-1.7 &micro;l/ml (mean: 1.2&micro;l/ml), Cuminum cyminum EO respectively. The result showed that Zataria multiflora EO is more efficient than other EOs against Candida species. There are no significant differences between different Candida trains to the EOs. Also, there were no significant differences in the MICs of these EOs against the azoles- resistant and azoles- susceptible isolates. Conclusion: In this study, the anti-Candida effects of the six EO against both azoles- resistance and azoles-susceptible isolates were similar. Given the documented resistance of different Candida species to synthetic and chemical antifungals, these EOs are effective replacement treatments for cutaneous and mucosal Candida infections especially for resistant or recurrent cases