Identification of Candida species by molecular and mycological methods in pregnant women with vulvovaginal candidiasis

Background: Vaginal candidiasis is common in during pregnancy. It may lead to complications like abortions, premature birth, low birth weight, chorioamnionitis and fungal systemic neonatal infection. The aim of present study was identification of Candida species by mycological and molecular methods in pregnant women with vaginal candidiasis. Methods: This cross-sectional study was performed on 80 pregnant women with or without clinical symptoms of vulvovaginal candidiasis referred to Shahid Noorani Talesh Hospital, Gilan University of Medical Sciences, Iran, from April to December 2015 (8 months). All specimens were examined by direct microscopy and culture on CHROMagar Candida medium for isolation and differentiation of major clinical-significant Candida species (spp.). Cultured media were incubated at 35 &deg;C for 48 hours and evaluated based on color and number of grown colonies. If no growth was observed, the media were incubated for several additional days. Subcultures were done on Sabouraud dextrose agar (Merck, Germany) and Corn meal agar with Tween 80 media (Micromedia, Hungary) for further study. Identification of Candida spp. carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: In this study, vulvovaginal candidiasis was observed in 20 (25%) patients. Twenty-two isolates were obtained from culture of specimens on CHROMagar Candida medium (Paris, France). The most common isolated species was Candida albicans 16 (72.8%) and followed by Candida glabrata 5 (22.7%), Candida tropicalis 3 (13.6%) and Candida krusei 1 (4.5%) cases. Two patients had mixed infection with 2 different Candida species (C. albicans and C. glabrata) While using PCR-RFLP method, the Candida species were identified as 13 (59.1%) Candida albicans, 5 (22.7%) Candida glabra, 3 (13.6%) Candida tropicalis and 1 (4.5%) Candida krusei cases, respectively. In direct examination were seen yeast budding cells and pseudohyphae in 8 culture positive specimens. In the present study, results of conventional mycological method in differentiation of Candida spp. were consistent with molecular results in 80% of cases. There was also significant correlation between vulvovaginal candidiasis with clinical symptoms (P<0.0001), including diabetes mellitus (P<0.014), and taking antibacterial drugs (P<0.003) in pregnant women. Conclusion: PCR-RFLP was able to identify correctly the Candida spp. as a complementary method

Diagnosis of fungal keratitis in patients with corneal lesions at Amiralmomenin hospital in Rasht, Iran

Background: Keratomycosis is a fungal infection of the cornea which could be sight-threatening and even causes eye loss. Considering the high humidity and the dominance of agriculture as important predisposing factors of keratomycosis in north of Iran, this study was carried out for diagnosis of fungal keratitis in patients with corneal lesions in Rasht, Gilan province, Iran.&nbsp; Methods: This descriptive cross-sectional study was conducted from July 2015 to November 2016 on 56 patients with corneal lesion suspected to keratomycosis and referred to eye emergency ward of Amiralmomenin hospital, Rasht, Iran. Corneal scraping was performed in all cases and specimens were subjected to direct examination and culture. Only colonies grown in sites of corneal scraping inoculation were considered significant. Fungal isolates were identified according to their macroscopic features of colonies and microscopic characteristics in slide cultures. Data were analyzed in SPSS software, version 21 (IBM SPSS, Armonk, NY, USA) and P<0.05 was considered significant. Results: The patients included 42 (75%) men and 14 (25%) women with the mean age of 49.5 years (9 to 90 years). Positive culture was observed in 9 cases but, only in one of these patients direct examination was positive and fungal elements were seen in 10% KOH preparation. Though, fungal keratitis was confirmed in 9 (16%) patients including seven (77.8%) men and two (22.2%) women. The majority of cases (88.9%) had a history of corneal trauma with plants and they were mainly farmer. According to statistical analysis, there was a significant association between corneal trauma and keratomycosis (P=0.007). The most common etiologic agents were Fusarium spp. (n: 4, 44.4%), followed by Aspergillus flavus (n: 2, 22.2%), Penicillium sp. (n: 1, 11.1%), Acremonium sp. (n: 1, 11.1%), and Cladosporium sp. (n: 1, 11.1%) respectively. Conclusion: In the presence of sufficient predisposing factors such as corneal injuries caused by plants, keratomycosis could be caused by a variety of fungi. Furthermore, low sensitivity of direct examination in this study, revealed the necessity of culture in diagnosis of keratomycosis

Drug susceptibility pattern of candida species isolated from skin of neonates to fluconazole, amphotericin B and caspofungin

Background: Candida species are opportunistic yeasts that are capable of causing different infections and diseases among immunocompromised patients. Since Candida infections are major causes and frequent of septicemia in Neonatal Intensive Care Units (NICU), and they are associated with high morbidity and mortality rates, so obtaining adequate treatment seems necessary. Low birth weight preterm infants are especially vulnerable to these devastating infections. The aim of this study was to evaluate the drug susceptibility of Candida species colonized on the skin and mucous membrane of neonates to fluconazole, amphotericin B and caspofungin. Methods: This study was carried out in the laboratory of medical mycology and serology, School of Public Health, Tehran University of Medical Sciences for the period of 7 months from June 2016 to December 2016. In this descriptive cross-sectional study, 23 isolates of Candida species including Candida parapsilosis, Candida albicans, Candida tropicalis, Candida guilliermondii and Candida krusei were studied. These under study isolates were previously isolated from skin and mucous membranes of neonates in NICU of Imam Khomeini Hospital and Children's Medical Center were identified by PCR-RFLP (Polymerase chain reaction-restriction fragment length polymorphism). Evaluation of antifungal drug susceptibility including fluconazole, amphotericin B and caspofungin was carried out. Antifungal susceptibility test was done according to the standard protocol Clinical and Laboratory Standards Institute (CLSI M27-A3) that is specific to the yeast fungi. Statistical analysis was done by using T-test in SPSS version 22 (IBM, Armonk, NY, USA) and P<0.05 was considered statistically significant. Results: In this study, C. parapsilosis, C. albicans and C. tropicalis had the most sensitivity to fluconazole. Clinical Isolates of C. guilliermondii were also sensitive to fluconazole, but in C. krusei sensitivity was dose-dependent. All isolated species were sensitive to amphotericin B and caspofungin. Conclusion: According to the results, all isolated Candida species were more sensitive to amphotericin B and caspofungin than other antifungal drugs. In final conclusion, Finally, it is emphasized that antifungal susceptibility testing is necessary to prevent treatment failure or recurrence of disease