Isolation and molecular characterization of Cryptococcus species isolated from pigeon nests and Eucalyptus trees.

BACKGROUND AND PURPOSE: Cryptococcus species are pathogenic and non-pathogenic basidiomycete yeasts that are found widely in the environment. Based on phenotypic methods, this genus has many species; however, its taxonomy is presently being re-evaluated by modern techniques. The Cryptococcus species complex includes two sibling taxa of Cryptococcus neoformans and Cryptococcus gattii. We aimed to investigate the possible distribution of Cryptococcus species in pigeon nests and Eucalyptus trees in Ilam, Iran, using molecular techniques. MATERIALS AND METHODS: Two hundred and seventy-four specimens were collected from pigeon nests and Eucalyptus trees during 2016-2017. All the specimens were sub-cultured on Sabouraud Glucose Agar with chloramphenicol and bird seed agar. For molecular identification, the ITS15.8SITS2 rDNA region was amplified using the first and fourth internal transcribed spacer (ITS1 and ITS4, respectively) primers. The purified products were applied for cycle sequencing reactions in forward direction with ITS1 primer. The obtained results were analyzed with Chromas 2.3. RESULTS: Thirty-three out of 186 cultures (17.7%) and 11 out of 88 cultures (12.5%) were positive among pigeon nest and Eucalyptus tree specimens, respectively. Cryptococcus albidus (17.2%), C. albidus var. kuetzingii (3.4%), C. adeliensis (3.4%), C. uzbekistanensis (3.4%), and C. neoformans var. grubii (3.4%) were isolated from pigeon nests, and Cryptococcusadeliensis (25%) was the only Cryptococcus species isolated from Eucalyptus trees. CONCLUSION: The presence of pigeons and Eucalyptus trees in the vicinity of some particular places such as rest homes and hospitals should be considered as a risk factor for the immunocompromised population.

Phospholipase and proteinase activities of Candida spp. isolates from vulvovaginitis in Iran.

OBJECTIVE OF THE STUDY: This study aims to characterize phospholipase and proteinase activities of Candida isolates from 82 vulvovaginal candidiasis (VVC) and to study the relationship of these activities with vulvovaginitis. METHODS: Totally 82 Candida isolates from vagina samples of VVC patients were randomly collected over the period between September and December 2014 from hospitalized patients at the general hospitals of Lorestan province, Iran. Isolates were previously identified by conventional mycological methods. The phospholipase and proteinase activities were evaluated by Egg yolk agar, Tween 80 opacity medium and agar plate methods. RESULTS: The most common Candida species was identified Candida albicans (n=34, 41.5%), followed by Candida famata (n=13, 15.8%), Candida tropicalis (n=11, 13.4%), and Candida parapsilosis (n=9, 11%). The most phospholipase activity was observed in Candida colliculosa (40%), followed by C. famata (38.5%), and Candida krusei (33.3%). The findings revealed that the correlation between phospholipase production by Candida spp. and the presence of VVC was not found to be statistically significant (P=0.91). All Candida spp. exhibited considerable proteinase activity; so that 100% of C. colliculosa, C. parapsilosis, Candida kefyr, and Candida intermedia isolates produced high proteinase activity with Pz 4+ scores. There was a significant correlation between proteinase production by Candida spp. and the presence of VVC (P=0.009). CONCLUSION: The obtained findings revealed that Candida spp. isolates may produce both virulence factors, phospholipase and proteinase. Although the phospholipase production was only observed in <40% of the isolates; however there was a significant association between proteinase production by Candida spp. and VVC.

Evaluation of the antifungal activities of various extracts from Pistacia atlantica Desf.

BACKGROUND AND PURPOSE: Despite the availability of various treatments for fungal diseases, there are some limitations in the management of these conditions due to multiple treatment-related side-effects. The present study was designed to investigate the antifungal properties of different extracts from Pistacia atlantica Desf. MATERIALS AND METHODS: Different parts of P. atlantica (i.e., dried fruit, fresh fruit and dried leaf) were separately extracted via percolation method with 80% methanol and water. Gas chromatography/mass spectrometry (GC/MS) analysis was performed to determine the main constituents of leaf and fruit extracts from P. atlantica. In vitro anti-Candida activities of the extracts against Candida albicans, Candida glabrata and Saccharomyces cerevisiae were studied. For this purpose, the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) were determined, using broth microdilution method, according to the modified M27-A3 protocol on yeasts, proposed by the Clinical and Laboratory Standards Institute (CLSI). RESULTS: Based on GC/MS analysis, the main constituents of P. atlantica fruit extracts were ß-myrcene (41.4%), α-pinene (32.48%) and limonene (4.66%), respectively, whereas the major constituents of P. atlantica leaf extracts were trans-caryophyllene (15.18%), α-amorphene (8.1%) and neo-allo-ocimene (6.21%), respectively. As the findings indicated, all the constituents exhibited both fungistatic and fungicidal activities, with MICs ranging from 6.66 to 26.66 mg/mL and MFCs ranging from 13.3 to 37.3 mg/mL, respectively. Among the evaluated extracts, the methanolic fresh fruit extract of P. atlantica was significantly more effective than other extracts (P<0.05). CONCLUSION: Based on the findings of the present study, novel antifungal agents need to be developed, and use of P. atlantica should be promoted in the traditional treatment of Candida infections.

Evaluation of antifungal activities of the essential oil and various extracts of Nigella sativa and its main component, thymoquinone against pathogenic dermatophyte strains.

OBJECTIVE: Plant extracts and plant-derived compounds are valuable sources as folk medicine for the treatment and prevention of a wide range of diseases including infectious diseases. In the present study, the antifungal activities of the essential oil and various extracts Nigella sativa and its active principle, thymoquinone against Trichophyton mentagrophytes, Microsporum canis and Microsporum gypseum as pathogenic dermatophyte strains have been evaluated. In addition, the cytotoxic effects of N. sativa against murine macrophage cells were determined. MATERIALS AND METHODS: In this study, the antifungal activity was studied by disk diffusion method and assessment of minimum inhibitory concentration (MIC) of extracts using broth macrodilution method. In addition, the cytotoxic activity of N. sativa was evaluated by colorimetric assay (MTT). The components of the N. sativa essential oil were also identified by gas chromatography/mass spectroscopy (GC/MS) analysis. RESULTS: The results showed that the essential oil and various extracts of N. sativa particularly thymoquinone have potent antifungal effects on T. mentagrophytes, M. canis and M. gypseum as pathogenic dermatophyte strains. In the assessment of the cytotoxicity activity, it could be observed that N. sativa had no significant cytotoxicity in the murine macrophages at low concentrations. While, thymoquinone in comparison with essential oil and various extracts of N. sativa showed higher cytotoxicity on murine macrophage cells. In the GC/MS analysis, thymoquinone (42.4%), p-cymene (14.1%), carvacrol (10.3%) and longifolene (6.1%) were found to be the major components of N. sativa essential oil. CONCLUSION: The findings of this study suggest a first step in the search of new antidermatophytic drugs and aid the use of N. sativa seeds in the traditional medicine for dermatophytic infections.

The comparison of the efficacy of various fixatives on diverse staining methods of Giardia lamblia cyst.

The definite and exact diagnosis of protozoa is possible using high magnification objective lenses, provided that suitable stained smears are prepared. Therefore, the appropriateness of both fixative and staining methods to the species of parasite, which is the main objective of this study, is important. In this study, five various fixatives including (Merthiolate iodine formalin) MIF, (Sodium acetate-acetic acid formalin) SAF, (Polyvinyl alcohol) PVA, formalin and schaudinn and four types of stains including Hematoxylin I, Hematoxylin II, Trichrome and Carbol-fuchsin were prepared using standard procedures. After the smears of stool samples containing Giadria lamblia cyst were prepared and kept for 24 h in various fixatives, the study was carried out using the four above-mentioned stains by changing the ingredients and time as well as by repeating the experiments. After fixing and staining all the smears in identical conditions along with the implementing interferences in the staining process, the following results were eventually obtained considering the morphologic indexes and negative and positive scores (from 1 to 20): formalin with 17 scores in hematoxylin I staining, formalin and SAF with 15 and 14 scores, respectively in Hematoxylin II staining, MIF with 13 scores in Trichrome staining and SAF, PVA, MIF with 11.5, 11.5 and 11 scores, respectively in carbol-fuchsin staining were found to be the best fixatives. Hematoxylin I staining using formalin fixative with 17 scores showed the best result while the maximum score for Carbol-fuchsin staining was 11.5 showing a necessity for more expenditure, time and expert cooperation to reach ideal results.