In vitro antifungal susceptibilities and molecular typing of sequentially isolated clinical Cryptococcus neoformans strains from Croatia.

A collection of 48 clinical Cryptococcus neoformans isolates from Croatia was investigated retrospectively using in vitro antifungal susceptibility testing and molecular biological techniques to determine mating type and serotype by PCR and amplified fragment length polymorphism (AFLP) genotyping. These isolates were obtained from 15 patients: ten were human immunodeficiency virus (HIV)-negative (66.7 %) and five were HIV-positive (33.3 %). From five patients, only one isolate was available, whilst from the other ten patients, two to 11 isolates were isolated sequentially. Antifungal susceptibility was tested by a broth microdilution method. Serotype A (genotype AFLP1) and serotype D (genotype AFLP2) were both found in six patients (40 % each), and serotype AD (genotype AFLP3) in three (20.0 %) patients. Mating type α (n = 12; 80.0 %) predominated and α/a hybrids were identified in 20.0 % of patients diagnosed with cryptococcosis. Two AFLP genotypes of C. neoformans were isolated during a single episode from one patient. The in vitro antifungal MIC(90) and susceptibility ranges for C. neoformans isolates were 0.5 µg ml(-1) (range 0.031-0.5 µg ml(-1)) for amphotericin B, 4 µg ml(-1) (range 1-4 µg ml(-1)) for flucytosine and fluconazole, 0.25 µg ml(-1) (range 0.031-0.5 µg ml(-1)) for itraconazole and 0.062 µg ml(-1) (range 0.031-0.25 µg ml(-1)) for voriconazole.

Candida utilis candidaemia in neonatal patients.

In recent years, an evident rise in the frequency of candidaemia caused by non-albicans Candida species has been reported. In this paper we present three cases of clinically manifested candidaemia caused by Candida utilis in neonatal patients hospitalized in the same neonatal intensive care unit within a 6 month period. To the authors' knowledge, only two cases of C. utilis candidaemia have been reported in the literature to date, but neither of these involved newborns. Clinical resolution and elimination of C. utilis from the blood were achieved using liposomal amphotericin B or caspofungin in all patients.

[Proteinase excretion ability of Candida albicans species isolates in clinical specimens]. / Ispitivanje sposobnosti lucenja proteinaze izolata vrste Candida albicans izdvojenih iz klinickih uzoraka.

AIM: The aim of the study was to establish whether the Candida (C.) albicans species isolates from samples of the patient digestive, respiratory and genitourinary systems differed in the ability of proteinase excretion. METHODS: A total of 1009 isolates of the C. albicans species obtained from 1009 clinical specimens of the digestive, respiratory and genitourinary systems of 666 patients were examined. All samples were inoculated onto Sabourauds glucose agar and incubated aerobically at 37 degrees C for 3-7 days. Identification of C. albicans was done by standard and commercial tests. To test the proteinase excretion ability, we employed the Odds and Abbott method. The isolates were inoculated on a culture medium containing agar, glucose, vitamins and beef albumin fraction V (pH = 3.2), and stored for 7 days at 30 degrees C. Each isolate was tested twice, with the results read by the same person. Development of thready colonies in a milky-white field was considered as positive finding. Study results are shown in easy-to-consult tables. Distribution differences were assessed by chi2-test. RESULTS: Of 1009 C. albicans species isolates obtained from clinical specimens, 695 (68.9%) had the proteinase excreting ability. The presence of this enzyme was demonstrated in 72.7% of the species isolates obtained from samples of the digestive system, 65.8% of isolates from respiratory system and 59.6% of isolates from genitourinary system. Analysis of chi2-test results showed no statistically significant difference in the ability of C. albicans species isolates from specimens obtained from digestive, respiratory and genitourinary systems to excrete proteinase. DISCUSSION: The results of the present study are in agreement with the results of most other researchers reporting on the proteinase excreting ability to be demonstrated in 40%-80% of C. albicans isolates. In the present study, the ability to excrete proteinase was demonstrated in 68.9% of C. albicans species isolates obtained from clinical samples. CONCLUSION: The ability to excrete proteinase was demonstrated in 59.6%-72.7% of C. albicans species isolates obtained from the patient digestive, respiratory and genitourinary systems. Analysis of the results yielded no statistically significant difference in the proteinase excreting ability among the isolates obtained from digestive, respiratory and genitourinary systems.

Biotypes of Candida albicans isolated from cardiovascular system and skin surveillance cultures of hospitalized patients.

The aim of the study was to biotype 59 isolates of Candida (C.) albicans from cardiovascular system samples (blood and intravenous catheter) and 123 isolates of the same species from skin surveillance cultures (swabs of the armpit, groins and intravenous catheter insertion sites) of hospitalized patients using the Odds and Abbott biotyping method. Biotyping of 59 isolates of C. albicans taken from the cardiovascular system samples revealed the presence of 16 biotypes. Biotype 355 was the most common biotype, accounting for 35.6% of all biotype isolates from this system. Biotyping of 123 C. albicans isolates from skin surveillance cultures detected 21 biotypes. Biotype 355 was most common, accounting for 17.9% of all biotype isolates from these samples. The two systems had 10 biotypes in common: 355, 155, 257, 305, 105, 315, 300, 015, 157, and 345. These biotypes accounted for 88.3% and 81.4% of all C. albicans biotypes isolated from the cardiovascular system and skin surveillance cultures, respectively. Biotypes 355, 155, and 257 were the biotypes most frequently shared in isolates from the two systems. These biotypes accounted for 57.7% and 43.1% of all C. albicans biotypes isolated from the cardiovascular system and skin surveillance cultures, respectively.

Species distribution and frequency of isolation of yeasts and dermatophytes from toe webs of diabetic patients.

The paper identifies fungal species, looking at the incidence of fungal isolation and risk factors influencing the development of fungal infection and colonization of interdigital spaces of the feet in 509 diabetic outpatients. Using standard mycologic diagnostic methods, fungi were detected in toe webs of 122 (24%) diabetic patients. The finding of fungi was twice as common in interdigital spaces of one (85/16.7% of the patients) than both feet (37/7.3% of the patients). Yeasts were the most common isolates (95/18.7% of the patients), followed by dermatophytic moulds (24/4.7% of the patients), whereas coexistence of yeasts and dermatophytes was the most infrequent finding (3/0.6% of the patients). From toe webs, 24 fungal species, 21 yeast species belonging to nine genera (Candida, Rhodotorula, Cryptococcus, Trichosporon, Saccharomyces, Blastoschizomyces, Geotrichum, Debaryomyces, and Ustilago) as well as three species of dermatophytes of the genera Trichophyton and Epidermophyton were isolated. The most frequently isolated fungi were Candida parapsilosis (59/11.6% of the patients) and Trichophyton mentagrophytes (16/3.1% of the patients). Although there was no correlation between the incidence of toe web space colonization with yeasts and dermatophytosis with the criteria of patient sex and age, and duration of diabetes, the difference in the incidence according to type of diabetes was statistically significant. In non insulin dependent diabetes mellitus patients, the incidence of fungal isolation from toe webs was statistically significantly higher (30.1%) than in insulin dependent diabetes mellitus patients (19.8%).