Hafize Sav

Investigation of the relationship between virulence factors and genotype of Candida spp. isolated from blood cultures.

INTRODUCTION The aim of study was to investigate the virulence factors of phospholipase, proteinase, esterase production and biofilm formation in Candida species isolated from patients with candidemia, and to assess their relationship with Candida genotypes derived after repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting. METHODOLOGY Fifty-two strains were identified to species level according to conventional methods and sequencing. The DiversiLab system was used for the genotyping. Enzyme activities and biofilm formation were evaluated using microbiological methods. RESULTS The 52 strains were identified as follows: 29 C. parapsilosis, 19 C. albicans, 2 C. glabrata, and 2 C. tropicalis. Phospholipase and proteinase activities were observed to have statistically significant differences between C. albicans and non-albicans Candida (NAC) strains (p < 0.05), with C. albicans strains showing higher virulence. Rep-PCR revealed eight major genotypes (A-H).The 19 C. albicans and the 33 non-albicans Candida isolates yielded seven (A-G) and four (A, B, C, H) genotypes, respectively. C. albicans strains were not shown to have a predominant genotype and showed higher phospholipase and proteinase activitiy than did NAC, regardless of genotype. Genotype H (52%) was the predominant genotype for the NAC including 27 C. parapsilosis strains, but the majority of strains showed low virulence. CONCLUSIONS NAC species were the most common causative agent for candidemia. Genotyping showed low transmission of C. albicans strains, but transmission of C. parapsilosis was high. In candidemia, several Candida virulence factors may be responsible at the same time. However, different genotypes of Candida strains showed different virulence activity.

Investigation of possible virulence factors in Candida strains isolated from blood cultures

BACKGROUND The Candida species, which are one of the most common causes of nosocomial bloodstream infections, present with high mortality and morbidity rates. This study aims to investigate the production of esterase, phospholipase, proteinase, and biofilm formation ability of the Candida strains isolated from the blood cultures. MATERIALS AND METHODS Between June 2011 and July 2012, the Candida strains, which were isolated from blood cultures of a total of 50 patients, were studied. The esterase activity was analyzed in the Tween-80 agar, while phospholipase activity was studied in the egg yolk agar. The proteinase activity and biofilm formation were identified by using the petri dish method and microplate method, respectively. RESULTS Of 50 specimens obtained from individual patients, 17 (34%) were identified as C. albicans, 14 (28%) as C. glabrata, 9 (18%) as C. parapsilosis, 5 (10%) as C. krusei, 4 (8%) as C. kefyr, and 1 (2%) as C. tropicalis. The rate of proteinase, phospholipase, and esterase positivity was higher in the C. albicans isolates. Biofilm formation was the highest in the C. parapsilosis strains. CONCLUSIONS Higher rate of virulence factors in the most commonly isolated Candida species than other species indicates that these virulence factors play a crucial role in the pathogenesis.

Antifungal Activity of Propolis Against Yeasts Isolated From Blood Culture: In Vitro Evaluation

Background: Due to the failure of available antifungal agents in the treatment of candidemia and the toxic activities of these drugs, a lot of researches are being conducted to develop new nontoxic and effective antifungal agents for optimal control of fungal pathogens. The aim of this study is to evaluate the in vitro antifungal activity of propolis against yeasts isolated from the blood cultures of intensive care unit patients.

Conventional Morphology Versus PCR Sequencing, rep-PCR, and MALDI-TOF-MS for Identification of Clinical Aspergillus Isolates Collected Over a 2-Year Period in a University Hospital at Kayseri, Turkey.

Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep‐PCR, and matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) for the identification of Aspergillus strains.

Biofilm Formation and Resistance to Fungicides in Clinically Relevant Members of the Fungal Genus Fusarium

Clinically relevant members of the fungal genus, Fusarium, exhibit an extraordinary genetic diversity and cause a wide spectrum of infections in both healthy individuals and immunocompromised patients. Generally, Fusarium species are intrinsically resistant to all systemic antifungals. We investigated whether the presence or absence of the ability to produce biofilms across and within Fusarium species complexes is linked to higher resistance against antifungals. A collection of 41 Fusarium strains, obtained from 38 patients with superficial and systemic infections, and three infected crops, were tested, including 25 species within the Fusarium fujikuroi species complex, 14 from the Fusarium solani species complex (FSSC), one Fusarium dimerum species complex, and one Fusarium oxysporum species complex isolate. Of all isolates tested, only seven strains from two species of FSSC, five F. petroliphilum and two F. keratoplasticum strains, recovered from blood, nail scrapings, and nasal biopsy samples, could produce biofilms under the tested conditions. In the liquid culture tested, sessile biofilm-forming Fusarium strains exhibited elevated minimum inhibitory concentrations (MICs) for amphotericin B, voriconazole, and posaconazole, compared to their planktonic counterparts, indicating that the ability to form biofilm may significantly increase resistance. Collectively, this suggests that once a surface adherent biofilm has been established, therapies designed to kill planktonic cells of Fusarium are ineffective.

The frequency, antifungal susceptibility and enzymatic profiles of Candida species in cases of onychomycosis infection

Although the frequency of candidal onychomycosis is increasing daily, there is little information in literature about the epidemiology, pathogenesis, and antifungal susceptibility of this dermatological disease. This study aimed to provide information about the epidemiology, pathogenesis, and azole susceptibility of Candida species isolated from patients living in a region with continental climate. After identification of the isolated strains using conventional methods, proteinase and phospholipase activities were determined by a plate method and biofilm-forming ability was determined using the microplate method. Susceptibility of the same species to fluconazole (FLU), voriconazole (VRC), miconazole (MNZ), itraconazole (ITZ), and ketoconazole (KTZ) were determined by microdilution method. The 50 Candida isolates included 23 C. parapsilosis (46%), 13 C. albicans (26%), 4 C. guilliermondii(8%), 4 C.tropicalis (8%), 2 C.krusei(2%), 1 C.lusitaniae (2%), 1 C. sake (2%), and 1 C. kefyr (2%) isolates. The geometric mean (GM) of the minimum inhibitory concentration (MIC) for FLU, KTZ, VRC, MNZ, and ITZ was 0.4 μg/mL, 0.08 μg/mL, 0.08 μg/mL, 0.2 μg/mL, and 0.6 μg/mL, respectively. Proteinase, phospholipase, and biofilm-forming ability were detected in 18%(9/50), 20%(10/50), and 6%(3/50) of the Candida isolates, respectively. We found that the most frequently isolated species is C.parapsilosis. On the basis of the GM values, the most effective azoles are ketoconazole and voriconazole. The isolated Candida species exhibited low phospholipase, proteinase, and biofilm formation activities.

Identification and molecular epidemiology of dermatophyte isolates by repetitive‐sequence‐PCR‐based DNA fingerprinting using the DiversiLab system in Turkey

Dermatophyte species, isolation and identification in clinical samples are still difficult and take a long time. The identification and molecular epidemiology of dermatophytes commonly isolated in a clinical laboratory in Turkey by repetitive sequence‐based PCR (rep‐PCR) were assessed by comparing the results with those of reference identification. A total of 44 dermatophytes isolated from various clinical specimens of 20 patients with superficial mycoses in Kayseri and 24 patients in Hatay were studied. The identification of dermatophyte isolates was based on the reference identification and rep‐PCR using the DiversiLab System (BioMerieux). The genotyping of dermatophyte isolates from different patients was determined by rep‐PCR. In the identification of dermatophyte isolates, agreement between rep‐PCR and conventional methods was 87.8 % ( 36 of 41). The dermatophyte strains belonged to four clones (A ‐D) which were determined by the use of rep‐PCR. The dermatophyte strains in Clone B, D showed identical patterns with respect to the region. In conclusion, rep‐PCR appears to be useful for evaluation of the identification and clonal relationships between Trichophyton rubrum species complex and Trichophyton mentagrophytes species complex isolates. The similarity and diversity of these isolates may be assessed according to different regions by rep‐PCR.