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.

[Comparison of Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth microdilution methods for determining the susceptibilities of Candida isolates]. / Candida izolatlarinin antifungal duyarliliginin belirlenmesinde Klinik Laboratuvar Standartlari Enstitüsü (CLSI) ve Avrupa Antimikrobiyal Duyarlilik Testleri Komitesi (EUCAST) sivi mikrodilüsyon yöntemlerinin karsilastirilmasi.

Candida species are among the top 10 pathogens causing bloodstream infections associated with high morbidity, mortality. In spite of the development of new antifungal drugs, epidemiological studies have shown that resistance to antifungal drugs among Candida isolates is becoming a serious problem. The aim of this study was to compare the antifungal broth microdilution methods of the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) for amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole and anidulafungin susceptibility of Candida blood isolates. The study consisted of 74 Candida albicans, 67 Candida parapsilosis, 30 Candida glabrata, and 18 Candida tropicalis isolates. The minimum inhibitory concentrations were determined after 24 and 48 hour of incubation with CLSI method and only 24 hour of incubation with EUCAST method except anidulofungin. The MIC values obtained by both methods were considered to be compatible within ± 2 dilution limits. The categorical agreement between methods for each antifungal agent was assessed using clinical break points and epidemiological cut-off values. The agreement (± 2 dilution) between the methods was found to be species, drug, and incubation time dependent. After 24 hour incubation, good agreement category (> 90%) was detected between amphotericin B, itraconazole, posaconazole and anidulofungin, but was lower category (< 85%) was determined with fluconazole and voriconazole especially for relatively slow growing C.glabrata and C.parapsilosis isolates. Excellent categorical agreement (100%) was observed for amfoterisin B/C.parapsilosis, C.glabrata, C.tropicalis and anidulofungin/C.albicans, C.glabrata, C.tropicalis but least category was determined for posaconazole and C.albicans (71.6% at 24 hour; 73% at 48 hour). In vitro resistance of therapeutically used fluconazole and anidulafungin determined by both methods was rare among C.albicans (1.3%, 2.7% respectively), C.glabrata (0%, 3.3% respectively) and C.tropicalis (0%, 5.6% respectively) isolates but, an increase of non-susceptible isolates were observed among C.parapsilosis (11.9% at 24 hour of incubation; 17.9% at 48 hour of incubation) for fluconazole. There was also a cross resistance between fluconazole and voriconazole for three C.parapsilosis isolates and one multidrug resistant (fluconazole, itraconazole, posaconazole and anidulofungin) C.albicans isolate (fluconazole, itraconazole, posaconazole and anidulofungin). As a result in this study, it was determined thatboth methods were similar and can be used according to preference of laboratories. The CLSI antifungal susceptibility test results can be assessed at the end of 24 hour incubation, but sometimes it is important that the evaluation should be performed as a result of 48 hour incubation in slow growing species such as C.glabrata.

[Evaluation of Two Methods (Nativ-Lugol Preparation and Enzyme-Linked Immunosorbent Assay) for Detection of Entamoeba histolytica in Stool Samples]. / Entamoeba histolytica Tanisinda Iki Metodun (Enzyme Linked Immunosorbent Assay (ELISA) ve Nativ-Lugol) Degerlendirilmesi.

OBJECTIVE: This study aims to compare the performance of Native-Lugol examination and EIA Antigen Detection Test using stool samples obtained from patients diagnosed as clinical gastroenteritis and submitted to the Parasitology Laboratory in Uludag University between January 2010 and February 2011. METHODS: The stool samples taken from 116 patients and sent to the laboratory of parasitology from various clinics including outpatient services have been investigated using Native-Lugol examination and EIA Antigen Detection Kit (Wampole® E. histolytica II Techlab®, Inc., Blacksburg, Virginia) methods on all the samples. RESULTS: In one of 116 stool samples (%0,86), E. histolytica/E. dispar cysts and/or trophozoites were detected by using direct microscobic (nativ-lugol) method. E. histolytica specific antigen was detected in 34 (29.3%) out of the sample set, and the patients were given adequate treatment. The highest rate of E. histolytica specific antigen positivity were observed in 11-19 age group. CONCLUSION: On account of the fact that the sensitivity of direct microscopy is quite low, it is concluded that, from the viewpoint of preventing the amebiasis suspected patients from false diagnosis and hence from receiving inadequate treatment, the use of the ELISA method is more appropriate and advantageous, as it is cost effective and does not require highly qualified staff.

Investigation of methicillin resistant Staphylococcus aureus in neonatal intensive care unit.

Methicillin resistant Staphylococcus aureus (MRSA) strains lead to severe infections in immunosupressive patients, geriatric population and premature infants. 27 MRSA strains isolated in the Neonatal Intensive Care Unit was considered as an outbreak and it was aimed to investigate the genetic and epidemiologic relation of the MRSA outbreak. MecA gene was investigated in the S. aureus strains and pulsed field gel electrophoresis (PFGE) was used to investigate the genetic relation between outbreak strains. MecA gene was showed in all isolates. PFGE revealed that there were two different strains and most of the isolates (25/27) were owing to same clone. One of the samples were found closely related with the common strain and the other sample was found genetically unrelated. To terminate the outbreak; liquid baby food was gained to the baby food kitchen, no more new patient was imported to the neonatal unit and none of the patients were exported from neonatal unit to other clinics during outbreak, education about infection control precautions was given to all the staff and nursing bottle dishwasher was obtained. To manage and terminate the outbreak, besides the infection control precautions, tests to determine the genetic relation between outbreak strains which are done in the microbiology laboratory are needed. Molecular analysis of outbreak strains will contribute to prove the epidemiologic and evolution of outbreaks.