Identification, molecular characterization, and antifungal susceptibility of Cyberlindnera fabianii strains isolated from urinary tract.

OBJECTIVES: Cyberlindnera fabianii is an opportunistic pathogen isolated from clinical specimens. It can be incorrectly identified as Candida utulis by phenotypic methods. This study aimed to accurately identify Cy.fabianii strains isolated from the urinary tract, and to determine their molecular characterization and antifungal susceptibilities as well. METHODS: Twenty-nine yeast strains isolated from urinary tract samples were studied. Strains were identified by phenotypically, sequence analysis and MALDI-TOF MS. Sequence analysis using different gene regions (ITS1-2,D1/D2,EF-1-alpha) in ribosomal DNA was performed for the molecular analysis. Phylogenetic analysis was done by the neighbor-joining method. Antifungal susceptibilities of strains were determined for nine antifungals by reference broth microdilution and the Sensititre YeastOne broth microdilution method (SensititreTMYeastOneTMAST Plate, Thermo Fisher Scientific™,USA) according to CLSI M60-Ed2 recommendations. RESULTS: All strains were identified as C.utulis phenotypically by conventional methods, however all strains were identified as Cy.fabianii by sequence analysis and MALDI-TOF MS. It was observed that the gene regions examined in terms of determining evolutionary relatedness did not show intraspecies nucleotide variations. In all strains, the MIC50/MIC90 values for fluconazole were higher than the other antifungals tested. CONCLUSION: Cy.fabianii should be considered in fluconazole-resistant urinary tract yeast infections. Although conventional phenotypical methods were insufficient to identify Cy.fabianii, it could be correctly identified with sequence analysis using different gene regions (ITS1-2,D1/D2,EF-1-alpha) in ribosomal DNA and MALDI-TOF MS.

Cryptococcal meningitis in a non-HIV patient with solid organ transplantation.

Cryptococcal meningitis (CM) is often associated with human immunodeficiency virus (HIV). Recently, this microorganism has been increasingly identified in HIV-negative patients. CM cases are encountered in HIV-negative individuals, especially secondary to liver disease, solid organ transplantation (SOT), tuberculosis, lymphoproliferative diseases with T-cell-mediated immunological disorders, long-term corticosteroid use, malignancies, diabetes mellitus, and sarcoidosis. Our patient is an HIV-negative, SOT case with CM. It should be considered that CM can also occur in HIV-negative patients. As in our case, patients receiving long-term immunosuppressive therapy should be evaluated for CM, and renal functions should be closely monitored during treatment. There is a need for more case reports on the subject, especially in CM detected HIV-negative patients, due to the different treatment protocols and challenging clinical conditions compared to HIV-positive cases.

Molecular epidemiology, virulence factors, and antifungal susceptibility of Candida inconspicua strains isolated from clinical samples in Turkey.

In this study, it was aimed to evaluate the molecular epidemiology, virulence factors, and antifungal susceptibility of clinical Candida inconspicua isolates. All isolates were identified by phenotypic methods and sequence analysis of ITS 1-2, D1/D2, EF-1 alpha. Proteinase, phospholipase, and esterase activities, biofilm formation, and antifungal susceptibilities were determined. All thirty isolates identified as Candida norvegensis by phenotypic methods were reidentified as C. inconspicua by sequence analysis, demonstrating the inadequacy of phenotypic methods to differentiate these 2 species. The gene regions examined in terms of determining evolutionary relatedness did not show intraspecies nucleotide variations. Therefore, different molecular approaches are needed to evaluate molecular epidemiology. Esterase, phospholipase, and biofilm formation were found to be positive in 100%, 100%, and 36.6% of the strains, respectively. The MIC50/MIC90 values for fluconazole and flucytosine were found to be higher than the other tested antifungals, which should be taken into account in the treatment.

Molecular epidemiology and antifungal susceptibilities of Aspergillus species isolated from patients with invasive aspergillosis.

OBJECTIVE: The aim of this study was to evaluate the demographic data, molecular epidemiology, and in vitro antifungal susceptibility results of patients with Aspergillus isolated from various clinical specimens. METHODS: A total of 44 Aspergillus strains were studied. The definition of invasive aspergillosis in patients was made according to European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. Strains were phenotypically and molecularly identified. Demographic characteristics of patients and genotypes of strains were evaluated. Phylogenetic analysis was done by the The Unweighted Pair-Group Method with Arithmetic Mean (UPGMA). Antifungal susceptibility of strains was determined according to The Clinical and Laboratory Standards Institute (CLSI)-M61-Ed2 and The European Committee on Antimicrobial Susceptibility Testing (EUCAST). RESULTS: A total of 11 patients were classified as proven and 33 as probable invasive aspergillosis. There was a statistically significant difference in age groups, subdisease, neutropenic, and receiving chemotherapy between groups. A total of 23 strains were identified as Aspergillus fumigatus, 12 as Aspergillus niger, 6 as Aspergillus flavus, and 3 as Aspergillus terreus. Phylogenetic analysis revealed five different genotypes. No statistical difference was found in the comparisons between patients groups and genotype groups. There was a statistically significant difference between genotype groups and voriconazole, posaconazole, and itraconazole Minimum Inhibition Concentration (MIC). CONCLUSION: Accurate identification of strains and antifungal susceptibility studies should be performed due to azole and amphotericin B resistance. Genotyping studies are important in infection control due to identifying sources of infection and transmission routes.

Molecular epidemiology and antifungal susceptibilities of Aspergillus species isolated from patients with invasive aspergillosis

SUMMARY OBJECTIVE: The aim of this study was to evaluate the demographic data, molecular epidemiology, and in vitro antifungal susceptibility results of patients with Aspergillus isolated from various clinical specimens. METHODS: A total of 44 Aspergillus strains were studied. The definition of invasive aspergillosis in patients was made according to European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. Strains were phenotypically and molecularly identified. Demographic characteristics of patients and genotypes of strains were evaluated. Phylogenetic analysis was done by the The Unweighted Pair-Group Method with Arithmetic Mean (UPGMA). Antifungal susceptibility of strains was determined according to The Clinical and Laboratory Standards Institute (CLSI)-M61-Ed2 and The European Committee on Antimicrobial Susceptibility Testing (EUCAST). RESULTS: A total of 11 patients were classified as proven and 33 as probable invasive aspergillosis. There was a statistically significant difference in age groups, subdisease, neutropenic, and receiving chemotherapy between groups. A total of 23 strains were identified as Aspergillus fumigatus, 12 as Aspergillus niger, 6 as Aspergillus flavus, and 3 as Aspergillus terreus. Phylogenetic analysis revealed five different genotypes. No statistical difference was found in the comparisons between patients groups and genotype groups. There was a statistically significant difference between genotype groups and voriconazole, posaconazole, and itraconazole Minimum Inhibition Concentration (MIC). CONCLUSION: Accurate identification of strains and antifungal susceptibility studies should be performed due to azole and amphotericin B resistance. Genotyping studies are important in infection control due to identifying sources of infection and transmission routes.

Stable and exacerbation period serum cytokine and periostin levels of the five distinct phenotypes of severe asthma.

BACKGROUND: The differences in molecular mechanisms during a stable period and the changes in the inflammatory responses during exacerbations between distinct severe asthma phenotypes remain unclear. In this study, we aimed to characterize stable and exacerbation period serum cytokine and periostin levels of 5 different predefined severe asthma phenotypes with real-life data. Changes in the viral infection-induced exacerbations were also analyzed. METHODS: Serum levels of 8 cytokines and periostin were measured from the sera obtained from the adult patients with five different severe asthma phenotypes based on the presence/absence of aeroallergen sensitivity, peripheral eosinophilia and chronic rhinosinusitis with nasal polyposis (CRSwNP) during stable and exacerbation periods, and from the matched controls. RESULTS: Serum IL-13, IL-25, TSLP, and periostin levels were similar between the patient and the control groups during stable and exacerbation periods. Serum IL-25 and TSLP levels, and peripheral eosinophil count and periostin level showed a strong correlation. Stable period periostin levels were significantly higher in eosinophilic patients, and eosinophilic patients without long-term systemic steroid therapy had higher IL-13 levels. Compared to stable period, exacerbation period serum periostin levels found significantly lower [5853 (2309-8427) pg/mL vs. 4479 (2766-6495) pg/mL; p = 0.05] and periostin levels were much lower in viral infection-induced exacerbations [2913 (893-4770) pg/mL vs. 7094 (4782-9596) pg/mL; p = 0.022]. DISCUSSION: Our study showed that serum periostin levels were decreased in viral infection-induced exacerbations and increased in the presence of eosinophilia independent from atopy and it can help to differentiate eosinophilia even if the patient is under long-term systemic steroid therapy. Also, serum IL-13 levels may reflect peripheral eosinophilia in patients without long-term systemic steroid use.

Studies on the Effectiveness of Ozone Therapy on the Treatment of Experimentally Induced Keratitis with Candida albicans in Rabbits.

PURPOSE: In the present study, antifungal activity of ozonated olive oil (OZO) and ozonated distilled water (ODW) in the treatment of experimentally induced keratitis with C. albicans in rabbits were investigated. METHODS: The Groups were composed of as 1, 2, 3, 4, 5, 7 (n = 5 rabbits, 10 eyes/in each group) and Group 6 (n = 10 rabbits, 20 eyes/in the group). Fourty-eight hours after C. albicans inoculation; Group 1 received fluconazle (FLU)+OZO drops, Group 2 received FLU drop, Group 3 received OZO drop, Group 4 received FLU+ODW drops, Group 5 received ODW drop, Group 6 (infected control group) and Group 7 received PBS drop (negative control group). Treatment continued in all groups for 22 days for every 8 hours. RESULTS: Cornea cultures made 24 days post inoculation revealed statistically significant differences (p < 0,05) with concern to C. albicans amounts between Group 6 and Group 1-5. Statistical comparison of corneal opacity and corneal ulcer and conjunctivitis values among the Group 6 and Group 1-5 were also different significantly (p < 0,05) on days 20 and 24 post inoculation. CONCLUSION: OZO and ODW were found to be effective in treating C. albicans keratitis in the present study. It has also been proven by this study that ODW contain 26 µg/ml was the most effective in the treatment of C. albicans keratitis.

Gamma-glutamyl transpeptidase-platelet ratio, systemic immune inflammation index, and system inflammation response index in invasive Aspergillosis.

OBJECTIVE: Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index are three systemic immune and inflammation indexes that were investigated for their diagnostic and prognostic proficiencies in cardiovascular diseases and cancers. However, their predictive values for invasive aspergillosis have not yet been studied. The aim of this study was to evaluate Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index levels and their diagnostic values in invasive aspergillosis. METHODS: A total of 23 patients with invasive aspergillosis and 23 sex- and age-matched healthy participants were included in this study. Complete blood count parameters and liver function tests were studied. Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index were calculated. RESULTS: Leukocyte, neutrophil, lymphocyte, and monocyte levels were statistically significantly higher in IA group (p=0.031, p=0.027, p=0.033, and p=0.001, respectively). In invasive aspergillosis group, platelets were numerically lower; Aspartate transaminase, alanine aminotransferase, and lactic dehydrogenase levels were numerically higher than those in control group but differences between levels were not statistically significant (p>0.05). The γ-glutamyl transpeptidase levels of patients were statistically significantly higher (p=0.007), and in addition, statistically significant differences were found between groups in terms of gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index (p<0.001, p=0.037, p=0.001, respectively). Receiver operating characteristic analysis was performed, and areas under the curves were evaluated. gamma-glutamyl transpeptidase-platelet ratio had the higher area under the curve than systemic immune inflammation index and system inflammation response index (AUC 0.849, 0.798, 0.693, respectively). The results from receiver operating characteristic analysis of the data suggested that the use of a cutoff value of 0.15 for gamma-glutamyl transpeptidase-platelet ratio would be optimum for clinical use to confirm independent predictors of patients with invasive aspergillosis. CONCLUSIONS: Gamma-glutamyl transpeptidase-platelet ratio is an independent, a useful predictor, and is superior to other evaluated markers in the diagnosis of inflammation in invasive aspergillosis. Gamma-glutamyl transpeptidase-platelet ratio may also be a helpful biomarker for clinicians to follow-up the inflammatory process of these patients.

Gamma-glutamyl transpeptidase-platelet ratio, systemic immune inflammation index, and system inflammation response index in invasive Aspergillosis

SUMMARY OBJECTIVE: Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index are three systemic immune and inflammation indexes that were investigated for their diagnostic and prognostic proficiencies in cardiovascular diseases and cancers. However, their predictive values for invasive aspergillosis have not yet been studied. The aim of this study was to evaluate Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index levels and their diagnostic values in invasive aspergillosis. METHODS: A total of 23 patients with invasive aspergillosis and 23 sex- and age-matched healthy participants were included in this study. Complete blood count parameters and liver function tests were studied. Gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index were calculated. RESULTS: Leukocyte, neutrophil, lymphocyte, and monocyte levels were statistically significantly higher in IA group (p=0.031, p=0.027, p=0.033, and p=0.001, respectively). In invasive aspergillosis group, platelets were numerically lower; Aspartate transaminase, alanine aminotransferase, and lactic dehydrogenase levels were numerically higher than those in control group but differences between levels were not statistically significant (p>0.05). The γ-glutamyl transpeptidase levels of patients were statistically significantly higher (p=0.007), and in addition, statistically significant differences were found between groups in terms of gamma-glutamyl transpeptidase-platelet ratio, system inflammation response index, and systemic immune inflammation index (p<0.001, p=0.037, p=0.001, respectively). Receiver operating characteristic analysis was performed, and areas under the curves were evaluated. gamma-glutamyl transpeptidase-platelet ratio had the higher area under the curve than systemic immune inflammation index and system inflammation response index (AUC 0.849, 0.798, 0.693, respectively). The results from receiver operating characteristic analysis of the data suggested that the use of a cutoff value of 0.15 for gamma-glutamyl transpeptidase-platelet ratio would be optimum for clinical use to confirm independent predictors of patients with invasive aspergillosis. CONCLUSIONS: Gamma-glutamyl transpeptidase-platelet ratio is an independent, a useful predictor, and is superior to other evaluated markers in the diagnosis of inflammation in invasive aspergillosis. Gamma-glutamyl transpeptidase-platelet ratio may also be a helpful biomarker for clinicians to follow-up the inflammatory process of these patients.

Stable and exacerbation period serum cytokine and periostin levels of the five distinct phenotypes of severe asthma.

BACKGROUND AND AIM: The differences in molecular mechanisms during stable period and the changes in the inflammatory responses during exacerbations between distinct severe asthma phenotypes remain unclear. In this study, we aimed to characterize stable and exacerbation period serum cytokine and periostin levels of 5 different pre-defined severe asthma phenotypes with real-life data. Changes in the viral infection-induced exacerbations were also analyzed. MATERIALS AND METHODS: Serum levels of 8 cytokines and periostin were measured from the sera obtained from the adult patients with five different severe asthma phenotypes based on the presence/absence of aeroallergen sensitivity, peripheral eosinophilia and chronic rhinosinusitis with nasal polyposis (CRSwNP) during stable and exacerbation periods, and from the matched controls. RESULTS: Serum IL-13, IL-25, TSLP and periostin levels were similar between the patient and the control groups during stable and exacerbation periods. Serum IL-25 and TSLP levels, and peripheral eosinophil count and periostin level showed a strong correlation. Stable period periostin levels were significantly higher in eosinophilic patients and eosinophilic patients without long-term systemic steroid therapy had higher IL-13 levels. Compared to stable period, exacerbation period serum periostin levels found significantly lower [5853 (2309-8427) pg/mL vs. 4479 (2766-6495) pg/mL; p=0.05] and periostin levels were much more lower in viral infection-induced exacerbations [2913 (893-4770) pg/mL vs. 7094 (4782-9596) pg/mL; p=0.022]. CONCLUSION: Our study showed that serum periostin levels were decreased in viral infection-induced exacerbations and increased in the presence of eosinophilia independent from atopy and it can help to differentiate eosinophilia even if the patient is under long-term systemic steroid therapy. Also, serum IL-13 levels may reflect peripheral eosinophilia in patients without long term systemic steroid use.

Recent Increase in the Prevalence of Fluconazole-Non-susceptible Candida tropicalis Blood Isolates in Turkey: Clinical Implication of Azole-Non-susceptible and Fluconazole Tolerant Phenotypes and Genotyping.

Candida tropicalis is the fourth leading cause of candidemia in Turkey. Although C. tropicalis isolates from 1997 to 2017 were characterized as fully susceptible to antifungals, the increasing global prevalence of azole-non-susceptible (ANS) C. tropicalis and the association between high fluconazole tolerance (HFT) and fluconazole therapeutic failure (FTF) prompted us to re-evaluate azole susceptibility of C. tropicalis in Turkey. In this study, 161 C. tropicalis blood isolates from seven clinical centers were identified by ITS rDNA sequencing, genotyped by multilocus microsatellite typing, and tested for susceptibility to five azoles, two echinocandins, and amphotericin B (AMB); antifungal resistance mechanisms were assessed by sequencing of ERG11 and FKS1 genes. The results indicated that C. tropicalis isolates, which belonged to 125 genotypes grouped into 11 clusters, were fully susceptible to echinocandins and AMB; however, 18.6% of them had the ANS phenotype but only two carried the ANS-conferring mutation (Y132F). HFT was recorded in 52 isolates, 10 of which were also ANS. Large proportions of patients infected with ANS and HFT isolates (89 and 40.7%, respectively) showed FTF. Patients infected with azole-susceptible or ANS isolates did not differ in mortality, which, however, was significantly lower for those infected with HFT isolates (P = 0.007). There were significant differences in mortality (P = 0.02), ANS (P = 0.012), and HFT (P = 0.007) among genotype clusters. The alarming increase in the prevalence of C. tropicalis blood isolates with ANS and HFT in Turkey and the notable FTF rate should be a matter of public health concern.

Evaluation of Candida colonization and use of the Candida Colonization Index in a paediatric Intensive Care Unit: a prospective observational study.

Invasive candidiasis is an important cause of morbidity and mortality, which primarily occurs in intensive care units. The Candida colonization index is an accepted score as an early warning tool for invasive candidiasis. This study was performed in a medical PICU with patients prone to contracting invasive candidiasis, to determine the usefulness of the Candida colonization index in forecasting invasive candidiasis in children. This prospective study including 87 patients (children 1 month to 16 years old with several illnesses and requiring ICU care) was conducted in a 22-bed medical PICU, Health Science University of Kayseri Training and Research Hospital, between January 2015 and September 2016. Those patients not on antifungal therapy, who were expected to stay more than seven days in PICU and had no history of a PICU stay within the previous two months were included in the study. In all patients, rectal, cervical, throat, axillary, perineal and nasal swab cultures, urine culture and blood culture tests were performed at admission and every week throughout their stay. Overall, 2639 swab and urine cultures (mean: 30.3) and 325 blood cultures (mean: 3.73) were obtained from 87 patients and a total of 576 grew Candida spp. In patients' swab and urine cultures C. albicans was detected in 64.5%, C. parapsilosis in 12.1%, C. glabrata in 7.5%, Saccharomyces spp in 3.0 %, C. tropicalis in 2.4%, C. krusei in 2.1% and C. kefyr in 1.2%. Three patients had C. albicans and one had C. parapsilosis growth in blood culture. Sensitivity, specificity, positive predictive value and negative predictive value for CI were found to be 33.73%, 100%, 6.7%, and 100%, respectively. Patients are at risk of fungal infection in paediatric intensive care units. Specificity and the negative predictive value of 100 % indicate that CI is a useful score to rule out the presence of invasive fungal disease. On the other hand, the low rate of sensitivity (33.3 %) and positive predictive value (6,7%) make this score less reliable in forecasting invasive candidiasis in children.

Cutaneous ulcerations caused by Paecilomyces variotii in a renal transplant recipient.

Skin infections caused by Paecilomyces species have been rarely described in patients with solid organ transplantation. Cutaneous manifestations are highly variable and include erythematous macules, nodules, pustules, and vesicular and necrotic lesions. The diagnosis of these infections is generally made by examination of a skin biopsy. Management of these fungal infections is difficult due to the immunocompromised state of the patients. Moreover, antifungal therapy and immunosuppressive drug interactions should be considered during treatment management. Herein, we reported a case of cellulitis caused by Paecilomyces variotii in a 56-year-old man who had undergone a kidney transplantation. Erythematous macular and nodular lesions on the left hand and left foot appeared first; within 2 months the skin lesions became ulcerated, hemorrhagic, and progressively painful and the patient was admitted to our hospital. The diagnosis was made by skin biopsy and tissue culture. The skin lesions resolved by the sixth week of the treatment with voriconazole.

[Fungemia and septic arthritis caused by Saprochaete capitata in a patient with fanconi aplastic anemia: a case report]. / Fanconi aplastik anemili bir hastada Saprochaete capitata'nin neden oldugu fungemi ve septik artrit: bir olgu sunumu.

Saprochaete capitata (formerly known as Blastoschizomyces capitatus, Trichosporon capitatum, Geotrichum capitatum) is a rare but emerging yeast-like fungus. It is commonly found in environmental sources and can be isolated from skin, gastrointestinal system and respiratory tract of healthy individuals as well. It mainly infects patients with hematological malignancies such as acute myeloid leukemia (AML), especially in the presence of neutropenia; and mortality rates are high in those patients. Although the data about the in vitro antifungal susceptibility are limited, it is being reported that amphotericin B and voriconazole are more effective on S.capitata isolates whereas caspofungin had no activity. Here, we report a case of fungemia and septic arthritis due to S.capitata in a patient with Fanconi aplastic anemia. A 22-year-old male patient with Fanconi aplastic anemia was hospitalized in our hematology department for bone marrow transplantation. Two days after the hospitalization, neutropenic fever developed and multiple nodules similar to candidiasis were detected in his liver with the whole abdomen magnetic resonance imaging (MRI). Caspofungin treatment (single 70 mg/kg loading dose, followed by 1 x 50 mg/kg/day) was started. The patient remained febrile, and his blood culture yielded S.capitata. The treatment regimen was changed to a combination of liposomal amphotericin B (3 mg/kg/day) and voriconazole (2 x 4 mg/kg/day). A few days later, pain and swelling came out on patient's left knee and he underwent a surgical process with the prediagnosis of septic arthritis. Culture of synovial fluid was also positive for S.capitata. On the 26th day of the hospitalization, the patient died due to sepsis and multiple organ failure. Patient's blood and synovial fluid samples were incubated in BacT/Alert automated blood culture system (bioMérieux, France). After receiving the growth signal, yeast cells were seen in Gram staining and cream-coloured, wrinkled, yeast-like colonies that were able to grow at 45oC and resistant to cycloheximide were detected on Sabouraud dextrose agar (SDA). Urease test was negative, and according to API 20C AUX (bioMérieux, France) system, none of the carbonhydrates were utilized except glucose. The isolates that were able to produce annelloconidia in corn meal-Tween 80 agar slide culture were identified as S.capitata. The identification was further confirmed by DNA sequence analysis. Minimal inhibitory concentrations (MICs) of amphotericin B, fluconazole, voriconazole, and caspofungin were found to be 0.5 µg/ml, 1.5 µg/ml, 0.032 µg/ml, and > 16 µg/ml respectively. Repetitive sequence based PCR (rep-PCR) (DiversiLab system, bioMérieux, France) was used to determine clonal relatedness of the isolates from blood and synovial fluid samples. The isolates were indistinguishable (similarity coefficient > 97%) according to rep-PCR. In conclusion, S.capitata infections should be taken into consideration in the presence of fungemia and septic arthritis in hematological patients who receive caspofungin therapy.

Utility of the Aspergillus galactomannan antigen testing for neutropenic paediatric patients.

Invasive aspergillosis (IA) is an increasingly important cause of morbidity and mortality particularly in paediatric patients. Early diagnosis and the initiation of efficacious antifungal treatments could affect the prognosis of these patients. The aim of this study was to determine the clinical contribution of Galactomannan (GM) screening in paediatric patients. We reviewed the records of all in-patients, and followed up, in the various units at the Medical Faculty Children's Hospital of Erciyes University (Kayseri, Turkey), those who had at least one GM assay result from August 2009 to April 2012. Paediatric patients were classified as proven, probable or possible, according to the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG). Twenty-five patients, with proven IA (n=3), probable IA (n=9) and possible IA (n=13) were included in the study. The GM antigen assay results were analysed in 158 blood samples from 47 patients. At the cut-off value of 0.5 ng/ml, the sensitivity was 68% [95% confidence interval (CI); 47-85]; specificity, 77% (95% CI; 55-92). To obtain more accurate results with GM testing, the diagnosis of IA should be confirmed by clinical investigation and the factors causing false positivity of the test should also be considered.

Genotypes and virulence factors of Candida species isolated from oralcavities of patients with type 2 diabetes mellitus.

BACKGROUND/AIM: This study compared the genotypes and virulence factors of Candida species isolated from oral cavities of healthy individuals and patients with diabetes mellitus (DM). MATERIALS AND METHODS: A total of 142 healthy individuals and 73 diabetic patients participated in this study. Study populations were classified into 4 groups as follows: Group I - Healthy, without caries; Group II - Healthy, with caries; Group III - DM, with caries; Group IV - DM, without caries. Diabetic patients' blood glucose and hemoglobin A1c concentrations were determined. Identification of Candida species was performed with conventional methods. Biofilm production, proteinase, phospholipase, and esterase were analyzed. The genetic diversity of Candida species was established using rep-PCR. RESULTS: The most isolated species was Candida albicans. There were statistical differences in terms of isolated Candida frequency between healthy subjects and diabetic patients. There was no statistical difference between the virulence factors of groups. Twelve genotypes were determined. While there were statistical differences in aerobe biofilm production, proteinase, and phospholipase activity between genotypes, there were no statistical differences in anaerobe biofilm production and esterase activity between genotypes. CONCLUSION: Diabetes has no effect on the activities of virulence factors of Candida species. Different genotypes of Candida albicans exhibited different virulence activities.

[Determination of Candida colonization and Candida score in patients in anesthesia intensive care unit]. / Anestezi yogun bakim ünitesinde yatan hastalarda Candida kolonizasyonu ve Candida skorunun belirlenmesi.

The colonization rate of Candida spp. reaches up to 80% in patients who reside in intensive care units (ICUs) more than a week, and the mean rate of development of invasive disease is 10% in colonized patients. Since invasive candidiasis (IC) in ICU patients presents with septic shock and high mortality rate, rapid diagnosis and treatment are crucial. The aim of this study was to assess the relationship between invasive infection and the determination of Candida colonization index (CI) and Candida score (CS) in patients admitted to ICU who are at high risk for IC and likely to benefit from early antifungal therapy. A total of 80 patients (34 female, 46 male; age range: 12-92 years, mean age: 69.57 ± 16.30) who were in ICU over seven days or longer of Anesthesia Department of Kayseri Education and Research Hospital between April, 2014 and July, 2015 were included in the study. None of the patients were neutropenic. After admission, throat, nose, skin (axillary region), urine, rectal swab and blood cultures have been collected weekly beginning from day zero. Isolation and identification of Candida strains were performed by using conventional mycological methods. CI was calculated as the ratio of the number of culture-positive distinct body sites (except blood culture) to the total number of body sites cultured. CI> 0.2 was considered as fungal colonization, while CI≥ 0.5 as intensive colonization. CS value was calculated according to the components including total parenteral nutrition (TPN) (plus 0.908 points), surgery (plus 0.907 points), colonization in multiple areas (plus 1.112) and severe sepsis (plus 2.038 points), and cut-off value for CS was accepted as >2.5. In our study, overall 1009 cultures (mean: 13 cultures per patient) were taken from 80 patients, and yeast growth was detected in 365 (36.2%) of them. Accordingly, among 68 (85%) of 80 patients included, in at least one sample, yeast growth was determined. No yeast growth was observed in the blood cultures. Of 365 yeast-positive cultures, C.albicans was isolated from 184 (50.4%), C.glabrata from 66 (18%), C.parapsilosis from 42 (11.5%), C.tropicalis from 12 (3.3%), C.kefyr from three (0.8%), and C.krusei from one (0.3%) samples, whereas six (1.6%) samples yielded other yeasts (3 Saprochaete capitata, 3 Trichosporon spp.) and 51 (13.9%) samples yielded multiple yeast growth. The highest colonization rates were detected in rectal swabs (27.4%), urine (23.3%) and throat (22.5%) samples. CI value was found as >0.2 in 65% (52/80), and ≥0.5 in 25% (20/80) of the patients, whereas CS value was >2.5 in only 2.5% (2/80) of the patients. In the statistical evaluation, significant correlations were found between fungal colonization (CI> 0.2) and gender (p=0.032) and length of stay in ICU (p=0.004), and between intensive colonization (CI≥ 0.5) and gender (p=0.008) and age (p=0.012). However, the correlation between Candida colonization and the presence of underlying diseases, APACHE II score, Glasgow coma scale, invasive procedures, use of extended-spectrum antibiotics, presence of bacterial infections, haemodialysis, transfusion and history of previous hospitalization was not statistically significant. Our results have also indicated a statistically significant relationship between fungal colonization and the positivity of C.albicans, C.glabrata, C.parapsilosis ang C.albicans/C.glabrata (p=0.001, p=0.002, p=0.008 and p=0.028, respectively), emphasizing the importance of species-level identification of Candida isolates. The reason of lacking of IC development in our patients may be explained by their low CI and CS values. In conclusion, monitoring of ICU patients who are at high risk for IC in terms of CI and CS would be beneficial. However it is clear that our data need to be supported by multi-center and high-scale studies.

Pulmonary infection caused by Talaromyces purpurogenus in a patient with multiple myeloma.

A 66-year-old female patient with multiple myeloma (MM) was admitted to the emergency service on 29.09.2014 with an inability to walk, and urinary and faecal incontinence. She had previously undergone autologous bone marrow transplantation (ABMT) twice. The patient was hospitalized at the Department of Haematology. Further investigations showed findings suggestive of a spinal mass at the T5-T6-T7 level, and a mass lesion in the iliac fossa. The mass lesion was resected and needle biopsy was performed during a colonoscopy. Examination of the specimens revealed plasmacytoma. The patient also had chronic obstructive pulmonary disease (COPD) and was suffering from respiratory distress. After consultation with an infectious diseases specialist the patient was placed on an intravenous antibiotherapy with piperacillin/tazobactam (4.5g x 3) on 17.10.2014. During piperacillin/tazobactam treatment, the patient suffered from drowsiness, her general condition deteriorated, and she had rales on auscultation of the lungs. The patient underwent thoracic computerized tomography (CT) which showed areas of focal consolidation in the lower lobes of the two lungs (more prominent on the left), and increased medullary density. The radiology report suggested that fungal infection could not be ruled out based on the CT images. The sputum sample was sent to the mycology laboratory and direct microscopic examination performed with Gram and Giemsa staining showed the presence of septate hyphae; therefore voriconazole was added to the treatment. Slow growing (at day 10), grey-greenish colonies and red pigment formation were observed in all culture media except cycloheximide-containing Sabouraud dextrose agar (SDA) medium. The isolate was initially considered to be Talaromyces marneffei. However, it was subsequently identified by DNA sequencing analysis as Talaromyces purpurogenus. The patient was discharged at her own wish, as she was willing to continue treatment in her hometown. Unfortunately, the patient died on December 8, 2014. In conclusion, apart from T. marneffei, less common strains such as T. purpurogenus should be considered when clinical samples obtained from patients with haematologic/oncologic disorders show fungal colonies that form red pigments on the culture media and when microscopic examination suggests a morphological appearance similar to Penicillium species.

[Evaluation of peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method in the identifi cation of Candida species isolated from blood cultures]. / Kan kültürlerinden izole edilen Candida türlerinin tanimlanmasinda peptid nükleik asit floresan in situ hibridizasyon (PNA FISH) yönteminin degerlendirilmesi.

In recent years, increased number of patients who are hospitalized in intensive care units, received immunosuppressive therapy and treated with broad-spectrum antibiotics that can lead an increase in the incidence of systemic candidiasis. In these patients, the most common clinical manifestation is candidemia. Since the identification of Candida species isolated from blood cultures is time consuming by conventional (morphological and biochemical) methods, rapid, reliable and accurate methods are needed. For this purpose novel systems have been developed to identify the agent directly. The aim of this study was to evaluate the peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method for the identification of Candida species by comparing with the conventional methods. A total of 50 patients who were admitted to Erciyes University Medical Faculty Hospital clinics and followed with prediagnosis of systemic fungal infections whose blood cultures were positive for the yeasts between July 2011 and July 2012 were included in the study. The conventional identification of Candida isolates was performed by considering macroscopic and microscopic morphology, germ tube test, cycloheximide sensitivity, urease activity and carbohydrate assimilation patterns with API 20C AUX (bioMerieux, France) test. PNA FISH method was conducted by the use of a commercial kit namely Yeast Traffic Light(®) PNA FISH (AdvanDx, USA). According to morphological and biochemical characteristics (conventional methods), 19 (38%) out of 50 Candida isolates were identified as C.albicans, 12 (24%) as C.glabrata, five (10%) as C.parapsilosis, five (10%) as C.kefyr, four (8%) as C.krusei, two (4%) as C.guilliermondii, two (4%) as C.tropicalis and one (2%) as C.lusitaniae. On the other hand, 24 (48%) of the isolates were identified as C.albicans/C.parapsilosis (with green fluorescence), 16 (32%) as C.glabrata/C.krusei (with red fluorescence) and one (%2) as C.tropicalis (with yellow fluorescence) properly, however one C.tropicalis strain was misidentified as C.albicans by PNA FISH method. Other eight (16%) strains which were not presented in the evaluation panel of PNA FISH kit (5 C.kefyr, 2 C.guillermondii and 1 C.lusitaniae), gave no fluorescence and determined as other Candida spp. According to this, when the species that could be detected with the kit (C.albicans, C.parapsilosis, C.glabrata, C.krusei and C.tropicalis) were considered, the concordance rate with the conventional methods was determined as 97.6% (41/42) and the total evaluation rate for all the species was 84% (41/50). In conclusion, the most frequent isolated species from blood cultures in our hospital was C.albicans, followed by C.glabrata and C.parapsilosis. Since PNA FISH testing is a practical, reliable and rapid (resulted in 90 minutes) method for the identification of Candida strains at species level isolated from blood cultures, it was thought to be useful in routine laboratories. However, further comparative studies are required with large number of strains with the consideration of cost-effectiveness.

Conventional Morphology Versus PCR Sequencing, rep-PCR, and MALDI-TOF-MS for Identification of Clinical Aspergillus Isolates Collected Over a 2-Year Period in a University Hospital at Kayseri, Turkey.

BACKGROUND: Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains. MATERIALS AND METHODS: A total of 24 consecutive clinical isolates of Aspergillus were collected during 2012-2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara. RESULTS: Samples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001). CONCLUSION: The good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required.