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Introduction. Polymicrobial infections including yeasts and bacteria are not rare and patients with polymicrobial bloodstream infection have higher early and overall case fatality rates. The diagnosis of invasive fungal and bacterial infections is mainly based on blood culture. Aim. The aim was to reveal the effect of concomitant bacteraemia on the detection of fungi from blood cultures in the presence of polymicrobial bloodstream infections involving Candida and non-Candida fungi and to show the superiority of blood culture bottles including selective fungal media in such situations. Methodology. Twenty-four polymicrobial bloodstream infection models – involving one fungus and one bacterium – were constituted by using clinical blood culture isolates ( Escherichia coli , Staphylococcus aureus , Pseudomonas aeruginosa , Candida albicans, Candida glabrata, Fusarium solani and Trichosporon asahii). The Plus Aerobic/F (PAF) and Mycosis IC/F (MICF) culture bottles were used with the BACTEC 9240 device. After a bottle signalled positive, direct microscopic examination and subcultures on agar plates were performed. Results. All of fungi that were inoculated alone and in combination were detected by both direct microscopic examination and subcultures on agar plates from MICF bottles, whereas direct microscopic examination only revealed the bacterial agents from PAF bottles including combinations. Furthermore, fungal growth was hidden by bacterial growth on blood agar subcultures from PAF bottles including combinations of F. solani, C. glabrata or T. asahii with bacteria. Conclusion. Blood culture bottles including selective fungal media that can allow selective growth of fungi and earlier detection of some species should be preferred in addition to non-selective blood culture bottles, especially in specific patient populations. Further, the use of selective agar plates such as inhibitory mould agar may contribute to the solution of this problem in clinical laboratories.

Introduction. The simultaneous use of antifungals with immunosuppressive agents has become a necessity for patients taking immunosuppressive therapy. However, antifungal drugs are problematic because of their limited target. Hypothesis. Scientists have been searching for new antifungals and some compounds with at least additive effects on antifungals. Calcineurin inhibitors used as immunosuppressive agents also attract attention due to their antifungal property. Aim. To evaluate the activity of two calcineurin inhibitors alone and in combination with amphotericin B (AMB), caspofungin (CAS), itraconazole (ITR), voriconazole (VOR) and fluconazole (FLU). Methodology. MICs of AMB, CAS, ITR, VOR, FLU and cyclosporine A (CsA) and tacrolimus (TAC) as calcineurin inhibitors were evaluated by the broth microdilution method against Candida albicans (n=13), C. krusei (n=7) and C. glabrata (n=10). Checkerboard and time-kill methods were performed to investigate the activity of combining calcineurin inhibitors with antifungal drugs. Results. The lowest MIC values were detected with VOR for all Candida isolates tested. Although we did not detect any inhibition for CsA or TAC alone at concentrations tested in this study, the combinations of CAS with CsA showed the highest synergistic activity (36.7%) by the checkerboard method, and CAS with CsA and ITR with TAC combinations exhibited apparent synergistic interaction by the time-kill method. However, the combinations of both CsA and TAC with AMB resulted in antagonistic interactions, especially against C. krusei isolate in time-kill testing. Conclusion. Synergistic interactions in the combinations of TAC or CsA with antifungal drugs, except for AMB, in many concentrations was found to be promising in terms of the treatment of patients with fungal infections.

Farnesol is an extracellular quorum-sensing molecule produced by Candida albicans. Farnesol is also a sesquiterpene alcohol existing in many herbal products and has various activity against fungal cells. We aimed to investigate the efficacy of farnesol alone and the contribution of farnesol on the activity of voriconazole and amphotericin B against Aspergillus clinical isolates in vitro. A total of 45 Aspergillus clinical isolates were used in this study. The MIC values of voriconazole, amphotericin B, and farnesol were determined using reference broth microdilution method. The interactions of farnesol with voriconazole and amphotericin B were investigated by the checkerboard method and evaluated based on the fractional inhibitor concentration index (FICI). The MIC ranges of farnesol, voriconazole, and amphotericin B were 1,500-6,000 μM, 0.125-1 μg/mL, and 0.125-0.5 μg/mL against Aspergillus fumigatus isolates, 3,000-12,000 μM, 0.125-0.5 μg/mL, and 0.25-2 μg/mL against Aspergillus flavus isolates, respectively. The most common interaction in combination tests was "no interaction," and synergistic interaction was not detected. The combinations of farnesol with voriconazole and amphotericin B had antagonistic activity against 38% and 27% of all isolates, respectively.We concluded that the responses of different fungal species against farnesol are variable, and different interactions may be observed when it is combined with different antifungals. Therefore, it should be noted that farnesol may have an adverse effect on some fungi or interact negatively with antifungals used in combination.

Purpose. Candida spp. are the most common causes of fungemia. Rapid and accurate diagnostic methods are very important for appropriate management of candidemia. At present, blood culture is the essential diagnostic test despite having a long detection time and low sensitivity rate. We aimed to investigate the ways to shorten the turnaround time from blood culture collection to final identification in candidemia. Methodology. Sixty clinical bloodstream isolates of Candida were included, and Plus Aerobic/F, Peds Plus/F and Mycosis IC/Fbottles were used with a BACTEC 9240 blood culture instrument. Germ tube production, carbohydrate assimilation (API 20C AUX) and peptide nucleic acid fluorescent in situ hybridization yeast traffic light tests were performed directly from positive-signalled bottles. Results. Time to positivity of blood cultures was affected by species of Candida, fungal load and bottle type. Candida tropicalis had the shortest and Candida glabrata had the longest time to positivity. Mycosis IC/F culture bottle had a significant superiority in the isolation of yeasts, especially for C. glabrata and if there was a low fungal load in the bottle. Direct germ tube test had 90 % sensitivity and 97.6 % specificity for Candida albicans in two hours after signalling. The compliance between direct and classical assimilation tests was 98.3 %. Sensitivity and specificity of peptide nucleic acid fluorescent in situ hybridization were 100 %. Conclusion. We think that it is possible to shorten the turnaround time for the identification of Candida in blood culture even with currently available methods.

WOS: 000485661700047, PubMed: 30980958, Objectives: This study investigated the antifungal resistance rates of isolates from candidaemia patients in 12 tertiary-care centres in Turkey. Methods: A total of 1991 Candida spp. isolates from 12 centres isolated from 1997-2017 were included in the study. Species/species complex (SC) identification was performed using conventional methods in all centres, occasionally accompanied by MALDI-TOF/MS. Antifungal susceptibility testing was performed for amphotericin B, fluconazole, itraconazole, posaconazole, voriconazole and micafungin (as echinocandin class representative) using the CLSI microdilution method. Resistance rates were determined according to CLSI clinical breakpoints (CBPs). For drugs and species with undetermined CBPs, epidemiological cut-off values were used for wild-type (WT)/non-WT categorisation. Results: No or low rates of resistance were detected in general for tested Candida spp. isolates. Specifically, overall resistance to fluconazole in isolates of Candida parapsilosis SC and Candida glabrata SC were 7.7% and 0.9%, respectively. Resistance rates for C. parapsilosis SC varied extensively from one center to other (0-47.1%). Importantly, no echinocandin resistance was detected. Rates of non-WT isolates were also generally low: fluconazole against Candida lusitaniae, 4.3%; posaconazole against C. parapsilosis SC, 3.5%; posaconazole against Candida krusei, 1.9%; and voriconazole against C. glabrata SC, 0.5%. Conclusion: This is the first multicentre report of antifungal resistance rates among candidaemia isolates in Turkey, suggesting low resistance rates in general. Due to varying rates of fluconazole resistance in C. parapsilosis SC isolates that was detected at remarkably high levels in some centres, further studies are warranted to explore the source, clonal relatedness and resistance mechanisms of the isolates. (C) 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved., PfizerPfizer [WS776070], This study was supported by an Investigator-Initiated Research grant from Pfizer [WS776070 to SA-A]. The sponsor had no role in the study design, data collection or analysis, or manuscript preparation and submission.

The sexual cycle of Candida glabrata is not known; however, genomic evidence is indicative of recombination among subpopulations and the genome harbours genes necessary for undergoing mating and meiosis, which may increase fitness. The relationship between specific mating type-like (MTL) loci and antifungal susceptibility is not well understood in C. glabrata. We investigated different combinations of clinical C. glabrata isolate mating types and their antifungal susceptibility profiles. Allele profiles of the mating genes of 103 clinical C. glabrata isolates were identified, and their antifungal susceptibility to azoles, echinocandins and amphotericin B were compared. The majority (88.3%) of screened isolates harboured the a allele in the locus. The MTL1, MTL2 and MTL3 loci harboured a (88.3%), a (95.1%), and ? (71.8%) alleles, respectively. The C. glabrata isolates were susceptible to echinocandins but displayed high minimal inhibitory concentrations (MICs) for azoles. The MIC ranges and MIC90 values of all isolates were 1.0 to ?64 and 8.0 µg mL-1 for fluconazole, 0.06 to ?16.0 and 0.5 µg mL-1 for voriconazole, 0.06 to ?16.0 and 1.0 µg mL-1 for posaconazole, ?0.015 to 0.06, and 0.03 µg mL-1 for caspofungin, ?0.015 to 0.06 and 0.015 µg mL-1 for anidulafungin and 0.5-2 and 2.0 µg mL-1 for amphotericin B, respectively. The mating gene alleles of the clinical C. glabrata isolates were not associated with differences in the MICs of the tested antifungals, except for the MTL3 ?-allele and echinocandins. The mating genotypes of the clinical C. glabrata isolates had no recognisable common effect on antifungal susceptibility. © 2019 Blackwell Verlag GmbH, This study was funded by the Research Fund of Mersin University

Rapid diagnosis and early treatment of invasive aspergillosis is crucial for the management of the patients with haematological malignancy. We evaluated 358 sera from 78 febrile neutropenic episodes in patient with invasive aspergillosis (IA) (one proven, 17 probable, and 60 possible) and 83 episodes in patients with no IA according to the EORTC/MSG criteria. Patient's specimens were tested by Mycassay Aspergillus PCR (first commercial real-time PCR test) and in house real-time PCR to investigate the presence of Aspergillus DNA, and by ELISA for detect the galactomannan (GM) antigen. We systematically investigated the medical background that can be effective on the test results. The hospitalisation period was longer in proven/probable episodes when compared with no IA (P = 0.001) and possible episodes. With regard to duration of neutropenia, the differences between both proven/probable with no IA (P = 0.023) and possible with no IA (P = 0.002) were highly significant. Similarly, the rates of T cell suppressant therapy in group proven/probable and possible episodes were significantly higher than in no IA (P = 0.005). There are significant differences in the performance of GM and PCR-based tests among studies, and standardisation is required. Therefore, it can be useful to determine the effective factors on these tests. The use of larger volume of sera improved the performance of real-time PCR for detection of Aspergillus DNA in high-risk adult patients in the present study. Some host factors such as duration of neutropenia and administration of T cell suppressants related to the development of IA.

PubMedID: 29725811 Candida parapsilosis sensu stricto is an emerging cause of hospital-acquired Candida infections, predominantly in southern Europe, South America, and Asia. We investigated the genetic diversity and antifungal susceptibility profile of 170 independent C. parapsilosis sensu stricto strains obtained from patients with candidemia who were treated at the Ege University Hospital in Izmir, Turkey, between 2006 and 2014. The identity of each strain was confirmed via PCR amplification and digestion of the secondary alcohol dehydrogenase-encoding gene. The 24-h geometric mean minimum inhibitory concentrations of the antifungal agents, in increasing order, were as follows: posaconazole, 0.10 µg/mL; voriconazole, 0.21 µg/mL; caspofungin, 0.38 µg/mL; amphotericin B, 0.61 µg/mL; anidulafungin, 0.68 µg/mL; and fluconazole, 2.95 µg/mL. Microsatellite genotyping of the isolates (using fluorescently labeled primers and a panel of four different short-nucleotide repeat fragments) identified 25, 17, 17, and 8 different allelic genotypes at the CP6, B5, CP4, and CP1 locus, respectively. Posaconazole, caspofungin, and amphotericin B showed the greatest in vitro activity of the tested systemic azole, echinocandin, and polyene agents, respectively, and the observed antifungal susceptibility of the isolates was shown to be independent of their isolation source. We obtained a combined discriminatory power of 0.99 with a total of 130 genotypes for 170 isolates tested. Finally, microsatellite profiling analysis confirmed the presence of identical genotype between separate isolates, supporting that effective surveillance and infection-prevention programs are essential to limit the impact of C. parapsilosis sensu stricto on hospitalized patients’ health. © 2018, Springer Science+Business Media B.V., part of Springer Nature. National Institutes of Health: BG 10 RM 11C106 National Institute of Allergy and Infectious Diseases S. Seyedmousavi (&) Molecular Microbiology Section, Laboratory of Clinical Immunology and Microbiology (LCIM), National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), BG 10 RM 11C106, 10 CENTER DR, 9000 Rockville Pike, Bethesda, MD 20892, USA e-mail: Seyedmousavi@nih.gov

Background Blood culture is the gold standard diagnostic method in candidemia in spite of long result time and low sensitivity rate. Early diagnosis is crucial for management of candidemia because a delay in treatment is related with increased mortality. We aimed to evaluate the direct applicability of antifungal susceptibility testing methods from positive blood culture bottles to save at least 24 hours. Methods Blood culture bottles were inoculated with 62 Candida isolates. Etest and broth microdilution (BMD) methods for six antifungals, disk diffusion (DD) method for two antifungals were performed, both directly from bottles and standardly. Results Essential agreements between direct and standard Etest methods were 87.1% for caspofungin and >90% for other antifungals, but the agreements of them with reference BMD were relatively low. Essential agreement between direct and standard BMD was >93%. Correlation between direct and standard DD methods was very high, negative correlations were observed between reference BMD and DD methods. Conclusion BMD is a reference method to evaluate the antifungal susceptibility, direct application of BMD might provide reliable results at least 24 hours earlier. Direct DD method may be a qualitative alternative. Direct susceptibility testing methods may be very useful to initiating the appropriate treatment on time.

Aspergillus fumigatus is an opportunistic pathogen responsible for life-threatening infections in immuncompromised patients. Data about the in vitro pharmacodynamics of antifungals against A. fumigatus are limited. In the present study, we investigated the fungicidal activities, at concentrations of 1, 4 and 16 times the minimum inhibitory concentration (MIC), of caspofungin (CAS), amphotericin B (AMB) and voriconazole (VORI) against eight A. fumigatus isolates through the use of time kill and 2,3-Bis [2-methoxy-4-nitro-5-(sulfenylamino) carbonyl-2H-tetrazolium-hydroxide] (XTT) reduction tests. By the conventional time kill test, AMB was fungicidal (≥99.9% reduction in colony forming units; CFU) for all isolates at 4-16 MICs after 48 h incubation. The fungicidal effect for VORI was determined at 4 × MIC for one isolate and at 16 × MIC for four isolates at 48 h of exposure. CAS was also fungicidal at 1 × MIC for one isolate and at 4-16 MICs for two isolates at 48 h. While the percentage of median killing of AMB was found by the time-kill method with XTT as 99% at 4 × MIC and 99.28% at 16 × MIC, that of VORI was 94.5% at 4 × MIC and 92.88% at 16 × MIC after 48 h of incubation. However, a significant increase was observed compared to initial inoculum size with CAS after 48 h. Since the XTT method measures all cellular viability in media, it may give more reliable results about pharmacodynamics of antifungal agents against Aspergillus spp. than the time kill test.

We retrospectively evaluated the distribution of clinical Candida spp. isolated over a 5-year period in our hospital relative to year, specimen types, hospital departments and their antifungal susceptibility patterns. Overall 3,756 Candida spp. were recovered from 10,857 specimens. In vitro antifungal susceptibility tests were conducted with 2,068 isolates against amphotericin B, fluconazole and itraconazole using the Etest method. C. albicans was isolated frequently from non-sterile body specimens while non-C. albicans Candida spp. were commonly recovered from sterile body specimens. Isolation rates of C. albicans were 83%, 61.2% and 49% in non-sterile body specimens, sterile body specimens and blood-sterile body fluids, respectively. C. krusei was an important isolate from specimens of patients in the Haematology and Bone Marrow Transplantation units and its rate of recovery increased in these departments. Amphotericin B resistance was detected in only seven C. krusei isolates, whereas 80% (n = 1,653), 76% (n = 1,572) and 99% (n = 2,061) of all isolates were susceptible to fluconazole, itraconazole and amphotericin B, respectively. In conclusion, the distribution of Candida species was variable among hospital departments and among body sites. These results may be useful in predicting potential fungal pathogens and the choice of antifungal treatment.

Candida glabrata is one of the most frequent organisms isolated from superficial and invasive fungal infections, after Candida albicans. This organism also exhibits intrinsically low susceptibility to azole antifungals and treatment often fails. The microdilution method is not very practical for use in routine susceptibility testing in the clinical laboratory, thus necessitating the use of other methods. In this study, we compared the in vitro activity of five antifungal agents in three different groups (echinocandin, polyene and azole) against 50 C. glabrata isolates by broth microdilution and disk diffusion methods recommended by Clinical Laboratory Standards Institute CLSI M27-A3 and CLSI M44-A, respectively. All the isolates were susceptible to amphotericin B (100%) and 98% of the isolates were susceptible to caspofungin by the broth microdilution method. Within the azole group drugs, voriconazole was the most active followed by fluconazole and itraconazole in vitro. The highest rate of resistance was obtained against itraconazole with a high number of isolates defined as susceptible-dose dependent or resistant. Although the disk diffusion method is easy to use in clinical laboratories, it shows very poor agreement with the reference method for fluconazole and itraconazole against C. glabrata (8% and 14%, respectively).

PubMedID: 26612621 Aspergillus species can cause ocular morbidity and blindness, and thus, appropriate antifungal therapy is needed. We investigated the in vitro activity of itraconazole, voriconazole, posaconazole, caspofungin, anidulafungin, and amphotericin B against 14 Aspergillus isolates obtained from patients with ocular mycoses, using the CLSI reference broth microdilution methodology. In addition, time-kill assays were performed, exposing each isolate separately to 1-, 4-, and 16-fold concentrations above the minimum inhibitory concentration (MIC) of each antifungal agent. A sigmoid maximum-effect (Emax) model was used to fit the time-kill curve data. The drug effect was further evaluated by measuring an increase/decrease in the killing rate of the tested isolates. The MICs of amphotericin B, itraconazole, voriconazole, and posaconazole were 0.5–1.0, 1.0, 0.5–1.0, and 0.25 µg/ml for A. brasiliensis, A. niger, and A. tubingensis isolates, respectively, and 2.0–4.0, 0.5, 1.0 for A. flavus, and 0.12–0.25 µg/ml for A. nomius isolates, respectively. A. calidoustus had the highest MIC range for the azoles (4.0–16.0 µg/ml) among all isolates tested. The minimum effective concentrations of caspofungin and anidulafungin were ?0.03–0.5 µg/ml and ?0.03 µg/ml for all isolates, respectively. Posaconazole demonstrated maximal killing rates (Emax = 0.63 h-1, r2 = 0.71) against 14 ocular Aspergillus isolates, followed by amphotericin B (Emax = 0.39 h-1, r2 = 0.87), voriconazole (Emax = 0.35 h-1, r2 = 0.098), and itraconazole (Emax = 0.01 h-1, r2 = 0.98). Overall, the antifungal susceptibility of the non-fumigatusAspergillus isolates tested was species and antifungal agent dependent. Analysis of the kinetic growth assays, along with consideration of the killing rates, revealed that posaconazole was the most effective antifungal against all of the isolates. © 2015, The Author(s). Astellas Pharma Gilead Sciences Seyedmojtaba Seyedmousavi has received research and travel grants from Astellas Pharma B.V. and travel grant from Gilead Sciences. All of the other authors have no competing interests. The authors alone are responsible for the content and composition of the manuscript.

WOS: 000379266000003, PubMed ID: 27249521, Introduction: Early detection of methicillin-resistant Staphylococcus aureus (MRSA) in colonized patients is very important for infection control procedures to prevent MRSA spread. We aimed to monitor MRSA carriage in intensive care unit (ICU) patients and to evaluate the speed and efficiency of conventional culture, immunological, chromogenic, and molecular methods together with genotyping. Methodology: Nasal and axillar swab specimens were obtained from patients in the ICUs of the general surgery and neurosurgery wards in a tertiary hospital once a week over four weeks between December 2009 and July 2010. Oxacillin and cefoxitin disk diffusion tests, oxacillin agar screening test, latex agglutination test, chromogenic agar, and real-time polymerase chain reaction (PCR) tests were used for isolation and identification of MRSA. MRSA isolates were typed using ribotyping and pulsed-field gel electrophoresis (PFGE) typing. Results: MRSA colonization was detected in 48 of 306 patients by real-time PCR. The MRSA colonization rate was 6.2%, 15.5%, and 38.5% at admission and in the first and second weeks, respectively. Sensitivity, specificity, positive and negative predictive values for all phenotypic tests were 98%, 99.6%, 98%, and 99.6%, respectively. The shortest handle time was observed in PCR. A total of three banding patterns were obtained from MRSA isolates by ribotyping, and PFGE analyses revealed 17 different pulsotypes varying from 11 to 18 distinct bands, showing high genetic diversity among the samples. Conclusion: Phenotypic MRSA screening tests in our study exhibited similar performances. The superiority of real-time PCR is its short turnaround time., Eskisehir Osmangazi University [200811030], This work was supported by a grant from Eskisehir Osmangazi University. Project no: 200811030.

Background Diabetes patients are particularly susceptible to fungal infections because their vascular and immunological systems are compromised. Objectives The present study aimed to determine prevalences of tinea pedis and onychomycosis, factors predisposing to their development, and antifungal susceptibilities of causative fungal species against fluconazole, itraconazole, and terbinafine in patients with type 2 diabetes mellitus (DM). Methods Study groups were defined according to hemoglobin A1C rates of ≥6.5% for the diabetes group and ≤5.7% for control subjects. A total of 600 diabetes subjects and 152 control subjects were evaluated. Rates of onychomycosis and tinea pedis in diabetes patients, and associations with age, gender, blood glucose level, duration of diabetes and serum lipid profile were investigated, as were the distribution and antifungal susceptibility of agents isolated. Results Patients with onychomycosis and/or tinea pedis numbered 85 in the diabetes group and nine in the control group (P = 0.006). The development of onychomycosis or tinea pedis was significantly related to increasing age and male gender. Although the most common agents were dermatophytes, non-dermatophyte fungal isolates were not uncommon. Terbinafine was the most effective drug against dermatophytes but was invalid for non-dermatophyte isolates by in vitro antifungal susceptibility testing. Conclusions The development of onychomycosis or tinea pedis was significantly related to type 2 DM, increasing age, and male gender. The most common isolate was Trichophyton rubrum. The isolation and identification of the fungus is important to the effective management of tinea pedis and onychomycosis in diabetes patients because non-dermatophyte fungi can cause these infections.

Summary Although conventional identification of pathogenic fungi is based on the combination of tests evaluating their morphological and biochemical characteristics, they can fail to identify the less common species or the differentiation of closely related species. In addition these tests are time consuming, labour-intensive and require experienced personnel. We evaluated the feasibility and sufficiency of DNA extraction by Whatman FTA filter matrix technology and DNA sequencing of D1–D2 region of the large ribosomal subunit gene for identification of clinical isolates of 21 yeast and 160 moulds in our clinical mycology laboratory. While the yeast isolates were identified at species level with 100% homology, 102 (63.75%) clinically important mould isolates were identified at species level, 56 (35%) isolates at genus level against fungal sequences existing in DNA databases and two (1.25%) isolates could not be identified. Consequently, Whatman FTA filter matrix technology was a useful method for extraction of fungal DNA; extremely rapid, practical and successful. Sequence analysis strategy of D1–D2 region of the large ribosomal subunit gene was found considerably sufficient in identification to genus level for the most clinical fungi. However, the identification to species level and especially discrimination of closely related species may require additional analysis.

Summary The incidence of invasive aspergillosis (IA) has increased over the last years, especially in immuncompromised patients with high mortality rates. Because of difficulties about the diagnosis; serological methods [galactomannan (GM) antigen test] and polymerase chain reaction (PCR) developed in recent years. MycAssay Aspergillus PCR performance in the diagnosis of IA was evaluated and compared with the GM and in-house PCR. This study was conducted with 358 serum samples obtained from 99 patient with febrile neutropenic episodes who were followed in haematology and bone marrow transplantation units. Patients were classified by the European Organization for the Research and Treatment of Cancer/Mycoses Study Group criteria, 18 of them is proven and probable IA. GM antigen test and two different real-time PCR; one of them is fist commercial PCR for IA; Mycassay Aspergillus and the other one is in-house real-time PCR performed. Sensitivity values were Mycassay Aspergillus PCR, in-house PCR, and GM 65.38%, 11.53% and 23.07%, respectively. The high sensitivity obtained from Mycassay Aspergillus PCR and sensitivity is increased by using a combination of diagnostic methods. GM antigen test and real-time PCR could be beneficial for early diagnosis and treatment of IA. For routine usage of PCR as diagnostic assay more studies needed in future.

Thermoascus crustaceus is a thermophilic fungus and the teleomorph form of Paecilomyces crustaceus. Thermoascus spp. have been rarely isolated from human mycoses as etiological fungal agents. We believe that our patient is the first case of catheter colonization with Thermoascus crustaceus. In a 50-year-old male patient undergoing chronic peritoneal dialysis, the mold was isolated from three separate, consecutive dialysate fluid specimens and peritoneal catheter tip. The patient had slight clinical findings and he was treated by early catheter removal without antifungal treatment. Therefore this case was considered as the colonization of the peritoneal catheter rather than peritonitis. Consequently, we think that the human pathogen fungal spectrum will continue to enlarge.

Since C. dubliniensis is similar to C. albicans phenotypically, it can be misidentified as C. albicans. We aimed to investigate the prevalence of C. dubliniensis among isolates previously identified as C. albicans in our stocks and to compare the phenotypic methods and DNA sequencing of D1/D2 region on the ribosomal large subunit (rLSU) gene. A total of 850 isolates included in this study. Phenotypic identification was performed based on germ tube formation, chlamydospore production, colony colors on chromogenic agar, inability of growth at 45 A degrees C and growth on hypertonic Sabouraud dextrose agar. Eighty isolates compatible with C. dubliniensis by at least one phenotypic test were included in the sequence analysis. Nested PCR amplification of D1/D2 region of the rLSU gene was performed after the fungal DNA extraction by Whatman FTA filter paper technology. The sequencing analysis of PCR products carried out by an automated capillary gel electrophoresis device. The rate of C. dubliniensis was 2.35 % (n = 20) among isolates previously described as C. albicans. Consequently, none of the phenotypic tests provided satisfactory performance alone in our study, and molecular methods required special equipment and high cost. Thus, at least two phenotypic methods can be used for identification of C. dubliniensis, and molecular methods can be used for confirmation.

OBJECTIVE The aim is to emphasise the importance for intestinal parasites' diagnosis by direct microscopic examination and the discrepancies among results from different microscopists. METHODS Stool specimens were obtained from 225 children with diarrhoea after the macroscopic examination, prepared by formalineether sedimentation methods and included in microscopically examination by three different independent investigators (parasitologist, microbiologist, research assistant). Furthermore, specimens were stained with the modified Ehrlich Ziehl Neelsen method and evaluated for Cryptosporidium and Cyclospora. RESULTS A total 161 specimens were evaluated as negative by all investigators. The number of specimens containing parasites detected by at least one investigator was 64; Cryptosporidium parvum 30, Blastocytis hominis 16, Endolimax nana 5, Giardia intestinalis 4, Dientamoeba fragilis 3, Ascaris lumbricoides 3, Entamoeba histolytica/dispar 2, Cyclospora cayetanensis 1. The concordance among investigators was observed for 21 (33%) specimens; when specimens were evaluated for the presence of leukocytes and/or parasites, concordance was detected for 58 (91%) of the 64 specimens. In particular, significant differences were observed for the species level identification. CONCLUSION Different results can be obtained by microscopic examination according to the experience and educational level of microscopists. Therefore, we think that these tests should be performed by persons who have sufficient education and experience, if possible, combined with at least two different methods.

We evaluated the postantifungal effects (PAFEs) of caspofungin (CAS), voriconazole (VOR), amphotericin B (AmB), and the combinations of CAS + VOR and CAS + AmB against 30 clinical Candida krusei isolates at 0.25, 1 and 4 times the MIC of each individually and in the indicated combinations. Antifungals were removed after 1 hour and colony counts were performed at 0, 2, 6, 24, and 48 h. VOR did not display any measurable PAFE regardless of antifungal concentrations, while AmB and CAS exhibited dose-dependent PAFE. The most effective agent producing a prolonged PAFE in this study was CAS. Although the combination of CAS with VOR generated longer PAFEs at 0.25 and 1 times their respective MICs in comparison with CAS alone, this combination was indifferent rather than synergistic. However, the combination of CAS with AmB at 4 times their MICs exhibited the best performance, reducing the colony counts during the 48 h after removal of drugs and resulted in synergic interaction in respect to 20 (67%) isolates. Consequently, CAS has a prolonged PAFE in vitro against C. krusei isolates, and the combination of AmB + CAS may increase significantly the efficacy of CAS. Our data may be useful in optimizing dosing regimens for these agents and their combinations, although further studies are needed to explore the clinical usefulness of our results.

Treatment of invasive Candida krusei infections can be difficult due to its intrinsic fluconazole resistance and its reduced susceptibility to amphotericin B and flucytosine. Caspofungin (CAS) acts on a different cellular target, and its combination with voriconazole (VOR) or amphotericin B (AmB) appears promising. We evaluated the activity of CAS, VOR and AmB alone and in combination at 1/4, 1, 4xMIC concentrations by time-kill method against 30 C. krusei isolates. All isolates were susceptible to CAS and VOR; AmB MICs were 2 mu g/ml for 50% of isolates by broth microdilution. CAS showed a fast killing activity at all concentrations; it was fungistatic at 1/4xMICs and fungicidal at 1-4xMICs in general. VOR displayed a concentration-independent fungistatic activity against all isolates. AmB exhibited a concentration-dependent activity; it was fungistatic at 1/4-1xMIC and fungicidal at 4xMIC. The most common interaction was indifference for both combinations. Frequency of synergic interaction for the VOR + CAS combination was 66.7% at 1/4xMIC after 48 h. The best results for CAS + AmB combination were obtained at 4xMIC in the first 4-8 h; synergic interaction was detected for 20 isolates (66.7%) at 4xMIC after 4 h. Consequently, VOR and CAS alone have been found effective, and high AmB MICs are remarkable against clinical C. krusei isolates in vitro. The combinations of CAS with VOR or AmB have exhibited promising results.

The aim of this study was to determine the frequency of tinea pedis and manuum (dermatophyte infections of the hands and feet) in adults in rural areas of Turkey, the risk factors and self-administered treatment options. A total of 2,574 people living in a rural area were enrolled in the study. Participants were asked demographic data, hygienic habits in a questionnaire. KOH preparations and culture were performed from suspicious lesions. Medical and alternative therapy methods and former dermatophytosis diagnosis history were taken from the respondents with suspicious lesions. Microbiological samples were taken from 285 (11.1%) participants. Culture was positive in 109 (4.2%) of those. The most common agent was Trichophyton rubrum. The predisposing factors were found as age older than 40, male gender and obesity. Forty-nine (44.9%) of patients had taken a medical therapy, 56 (51.4%) had performed non-medical methods (cologne, Lawsonia inermis-Henna and softener creams). Patient's education about the treatment compliance is important.

The aim of this study was to investigate the in vitro interaction of amphotericin B in combination with fluconazole or voriconazole against Candida albicans clinical isolates by using a broth microdilution checkerboard assay and E-test. A total of 30 C. albicans strains isolated from blood, urine, sputum and pus samples were included to the study and the minimum inhibitory concentrations (MICs) of amphotericin B (AMB), fluconazole (FLU) and voriconazole (VOR) were determined by broth microdilution method and E-test. All strains tested for susceptibility were interpreted as susceptible by both methods (FLU MICs8 microg/ml, VOR and AMB MICs1 microg/ml). The rates of MIC agreement between two methods were as follows: AMB, 83%; FLU, 97%; VOR, 97%. AMB+ FLU and AMB+VOR combinations were tested by checkerboard broth microdilution and E-test methods. The combination test results were determined by using the fractional inhibitory concentration (FIC) index as synergistic, indifferent or antagonistic. AMB+FLU combination tested by checkerboard broth microdilution revealed synergy in one strain (3.3%) and antagonism in none, while the same combination tested by E-test revealed synergy in two (6.6%) and antagonism in four (13.3%) strains. The strains which exhibited synergy were different from eachother in two assays. This combination led to indifferent results in 23 (76.6%) of the strains. On the other hand AMB+VOR combination yielded synergistic results in two (6.6%) strains by both of the methods, however, these two strains were again different from eachother. No antagonism was detected by AMB+VOR combination while the combination was indifferent in 26 (86.6%) of the strains. Agreement between the checkerboard and E-test results was 87%. Although significant synergy was not detected in AMB+azole combinations, it was yet hopeful to obtain no antagonism. However, multi-center, large-scale, well standardized in-vitro and clinical studies about AMB and azole interaction which is a matter of debate, are necessary.

The relationship between the airborne contaminants obtained from operating theatres and intensive care units and the colonizing and infecting microorganisms isolated from patients were investigated. Air samples were obtained with the biocollector air IDEAL (BioMerieux, France). During the study period (19 weeks), a total of 77 air samples and 870 clinical specimens (swabs from throat, nose, conjunctiva and skin) from 174 patients were collected weekly. Microorganisms were identified by using Vitek system (BioMerieux, France) and conventional methods. According to the criteria of Federal Standard 209E (FD 209E) on cleanrooms, the conventionally ventilated operating- and general surgery rooms, and the anesthesia intensive care unit have been ranked as less than class 3.5 and 3, respectively. The frequency of nosocomial infection related to air-colonization was higher in patients of anestesia intensive care unit (16.4%), than in those of general surgery intensive care unit (4.9%). In general surgery rooms and anesthesia intensive care unit, the most frequent air-colonization related nosocomial infections were surgical wound infections and bacteremia, respectively. The most frequently isolated microorganisms were methicillin resistant Staphylococcus aureus (MRSA) and Acinetobacter baumannii. It can be concluded that, total number of airborne viable particles in the critical areas such as operating theatres and intensive care units, seems to be a significant risk factor for the development of nosocomial infections in immunocompromised patients.