Frequency of azole resistance in clinical and environmental strains of Aspergillus fumigatus in Turkey: a multicentre study.

OBJECTIVES: Aspergillus fumigatus causes several diseases in humans and azole resistance in A. fumigatus strains is an important issue. The aim of this multicentre epidemiological study was to investigate the prevalence of azole resistance in clinical and environmental A. fumigatus isolates in Turkey. METHODS: Twenty-one centres participated in this study from 1 May 2018 to 1 October 2019. One participant from each centre was asked to collect environmental and clinical A. fumigatus isolates. Azole resistance was screened for using EUCAST agar screening methodology (EUCAST E.DEF 10.1) and was confirmed by the EUCAST E.DEF 9.3 reference microdilution method. Isolates with a phenotypic resistance pattern were sequenced for the cyp51A gene and microsatellite genotyping was used to determine the genetic relationships between the resistant strains. RESULTS: In total, resistance was found in 1.3% of the strains that were isolated from environmental samples and 3.3% of the strains that were isolated from clinical samples. Mutations in the cyp51A gene were detected in 9 (47.4%) of the 19 azole-resistant isolates, all of which were found to be TR34/L98H mutations. Microsatellite genotyping clearly differentiated the strains with the TR34/L98H mutation in the cyp51A gene from the strains with no mutation in this gene. CONCLUSIONS: The rate of observed azole resistance of A. fumigatus isolates was low in this study, but the fact that more than half of the examined strains had the wild-type cyp51A gene supports the idea that other mechanisms of resistance are gradually increasing.

[Antifungal susceptibilities of Aspergillus spp. strains isolated from invasive aspergillosis cases]. / Invazif aspergillozlu olgulardan izole edilen Aspergillus spp. suslarinin antifungal duyarliliklari.

Aspergillus species found abundantly in the outer environment and hospital setting may lead to serious morbidity and mortality particularly in patients with suppressed immunity. This retrospective study was aimed to investigate the antifungal susceptibilities of Aspergillus spp. isolated from aspergillosis cases being hospitalized. Aspergillus spp. isolated from samples of the patients with suspected fungal infections between January of 2002 and October of 2007, were investigated. A total of 678 samples (420 lower respiratory tract, 202 sterile body fluids, and 56 biopsy/tissue specimens) from 569 patients were included in the study. The samples were incubated in 25 degrees C and 35 degrees C on brain-heart-infusion agar supplemented with blood and on Sabouraud dextrose agar. Gram and Giemsa stained samples were also examined by microscopy. Mold type of fungi were identified by conventional techniques. "Invasive aspergillosis" was described according to criteria of Invasive Fungal Infections Cooperative Group of the European Organization for Research and Treatment of Cancer. A. fumigatus (n = 8), A. flavus (n = 2) and A. niger (n = 2) were isolated from 12 patients' samples (2.1%), 9 of them were lower respiratory tract and one of each was ascid, brain biopsy and pleural fluid specimens. All of those patients have had an underlying diseases such as malignancy. The susceptibility of the isolates to caspofungin, voriconazole, itraconazole and amphotericin B was tested by broth microdilution susceptibility testing and to posaconazole by E-test (AB Biodisk, Sweden). The lowest minimum inhibitory concentration (MIC) (< or = 0.125 microg/ml) values were detected for caspofungin and posaconazole for Aspergillus spp., however, the highest MIC values were detected for amphotericin B (> 1 microg/ml). MIC values of the all strains except one, were detected as < or = 0.5 microg/ml for voriconazole and itraconazole. In one A. niger strain itraconazole MIC value was 2 microg/ml. Since the number of other species was low, MIC50 value was determined only for A. fumigatus strains and it was found that the highest MIC50 value was for amphotericin B (2 microg/ml) and the lowest MIC50 values were for posaconazole (0.064 microg/ml), caspofungin (0.064 microg/ml), itraconazol (0.25 microg/ml) and voriconazol (0.25 microg/ml). Since caspofungin and posaconazole revealed the lowest MIC values, they should be taken into consideration in choice of therapy of aspergillosis cases in our hospital.

[Investigation of the agents and risk factors of dermatophytosis: a hospital-based study]. / Dermatofitozlarda etkenlerin ve risk faktörlerinin arastirilmasi: hastane bazli bir Calisma.

The aims of this study were the detection of distribution of dermatophyte species isolated from the clinical samples of patients with dermatophytosis and the evaluation of risk factors for the development of dermatophytosis. A total of 441 skin, nail and scalp/hair specimens obtained from 301 patients (151 were male; age range 2 months-80 years, median 42 years) and 884 foot and hand skin and nail specimens obtained from 221 control subjects (110 were male; age range 5-75 years, median 36 years) were included to the study between the period of January to December 2005. All the samples have been evaluated by direct microscopic (DM) examination and by culture. A total of 121 (40.2%) patients yielded positivity for dermatophytes, of them 63 were positive by both DM and culture methods, seven were only culture positive, and 51 were only DM positive. Nine (9.8%) of 92 culture positive samples from 70 patients were found negative in DM, while 85 (50.6%) of 168 DM positive samples from 114 patients were negative in culture. 23.5% (12/51) of DM positive but culture negative patients were given antifungal therapy previously. The most prominent species isolated from the cultures were Trichophyton rubrum with a rate of 68.4% (63/92), followed by T. mentagrophytes (18.4%); T. violaceum (3.3%); T. verrucosum, T. tonsurans and Epidermophyton floccosum (2.2% for each); T. schoenleini, Microsporum canis and Trichophyton sp. (1.1% for each). Of the patient samples whose cultures were positive, 45% were from the foot skin. The presence rate of dermatophytes in controls was found as 3.2% (7/221); T. rubrum was isolated from the foot skin of five and T. mentagrophytes was isolated in toenail of two control subjects. About 42% of the samples belonged to the patients who admitted to hospital between December to February period. The evaluation of the risk factors revealed that presence of trauma, pet contact, ritual cleansing and diabetes mellitus had no effect on the development of dermatophytoses, however the presence of fungal infection in the family, male gender, some professions (being farmer, worker and retired), and the use of immunosupressive drugs have been found to increase the risk of dermatophytosis. The number of cases with dermatophytoses started to increase beginning from the age of 20 and peaked in the ages between 40-59 years old. As a result T. rubrum was determined as the most frequently isolated dermatophyte and tinea pedis was the most frequently observed clinical form in our hospital, emphasizing the importance of early diagnosis and effective treatment in superficial fungal infections which have high morbidity.

[Aspergillus spp. isolations from respiratory tract samples in Trakya University Hospital]. / Trakya Universitesi Hastanesi'nde solunum yolu örneklerinde Aspergillus spp. izolasyonlari.

The characteristics of cases diagnosed as aspergillosis and Aspergillus spp. strains isolated from the respiratory tract samples in Mycology Laboratory of Trakya University Hospital between January 2002 and May 2006 were investigated. In this period, 137 bronchoalveolar lavages, 95 sputum, nine tracheal aspirates, three lung biopsies and one bronchial biopsy of 85 patients were processed. The samples were incubated in 25 degrees C and 35 degrees C media by culturing on brain heart infusion agar with blood and Sabouraud dextrose agar. Presence of leucocytes and fungal structures were searched in the smear stained by Gram and Giemsa. The patient was defined as probable aspergillosis case, if he/she patient had clinical findings, lung infiltration or fungus ball radiologically, at least one risk factor predisposing to aspergillosis and isolation of Aspergillus spp. in lower respiratory tract samples without finding of other nonmycotic infection. Of 22 patients isolated Aspergillus spp., 13, six, two, one were internalized in chest diseases, haemotology, neurosurgery and oncology clinics, respectively. Seven positive cultures were considered as findings of aspergillosis. Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger were isolated in three, two, and two patients, respectively. Fungal structures were detected in only one sample in the direct microscopical examination. Ages of seven patients, five were males and two were females, were between 15 and 60. Predisposing risk factors were acute leukemia in six patients and lung cancer in one patient. Five patients were neutropenic and one was neutrophylic. Fungus ball was detected in radiological imaging of one patient, had a pulmonary cavitary lesion. Conventional amphotericine B was used in their therapies. Antifungal agents were switched to caspofungin and itraconazole in two and one patients, respectively. Three patients died in four weeks after isolation of Aspergillus spp. Aspergillosis cases were not high in our hospital because of absence of transplantation center for bone marrow or solid organ.

A Case Report on Aspergillus lentulus Pneumonia.

BACKGROUND: Aspergillus lentulus was described as a new species in 2005 but it was isolated from Turkey for the first time. CASE REPORT: A. lentulus was isolated as the cause of pneumonia from a patient who had renal transplantation 4 months ago. The patient received immunosuppressive treatment after transplantation. A. lentulus was isolated from his sputum as an agent in pneumonia developed 4 months after the transplantation. Leukocytes, blastospores, and hyphae were seen in both Gram- and Giemsastained smears of the sputum. The isolate was identified by using the Maren A. Klich algorithm and molecular methods and confirmed by the reference laboratory of the CBS Fungal Biodiversity Centre (The Netherlands). In the susceptibility tests of the isolate, minimal inhibitory concentrations for amphotericin B, voriconazole, posaconazole, and caspofungin were found to be 0.5 µg/mL, 0.25 µg/mL, 0.125 µg/mL, and 0.25 µg/mL, respectively. The patient recovered with voriconazole treatment (2×200 mg/day). CONCLUSION: The use of the molecular tests is important for identification of A. lentulus strains because they are very easily confused with A.fumigatus strains according to phenotypic characteristics.