[A comparison of the costs, reliability and time of result periods of widely used methods, new molecular methods and MALDI TOF-MS in the routine diagnosis of Candida strains]. / Candida tür tayininde rutinde yaygin olarak kullanilan yöntemlerle yeni kullanilmaya baslayan MALDI TOF-MS ve moleküler yöntemlerin sonuç verme süreleri, maliyetleri ve güvenilirliklerinin karsilastirilmasi.

In recent years, the fast and accurate identification of the Candida species is of great importance as the response to antifungal treatment differs among species. Following the treatment of several immunosuppressive diseases, fungal infections can emerge. The aim of this study was to compare the accuracy, costs and time of result periods of the methods used in the identification of the most common human fungal infectious agent, Candida strains. From various clinical samples sent to the Microbiology Laboratory of Karabuk University Training and Research Hospital between July 2016-December 2017, a total of 91 yeast isolates cultivated in blood agar (Becton Dickinson, USA) and/or Sabouraud dextrose agar (SDA-Oxoid, UK), confirmed with colony morphology and microscopic appearance, identified as Candida species with a fully automated identification system (Phoenix™ Yeast ID Panel, Becton Dickinson Diagnostics, USA) were included in the study. All the samples were examined with sequence analysis using ITS1 forward 5'-TCC GTA GGT GAA CCT GCG G-3' and ITS4 reverse 5'-TCC TCC GCT TAT TGA TAT GC-3' primers (Iontek, Turkey) and the matrix-assisted laser desorption-ionisation time of flight mass spectrometry (MALDI TOF-MS) systems. Molecular sequence analysis was accepted as the gold standard method and the results were compared with those of the other methods MALDI TOF-MS and Phoenix™ Yeast ID Panel in respect of the accuracy of the identification of Candida strains. According to the results of the DNA sequence analysis of the 91 Candida isolates included in the study, 24 were identified as Candida albicans, 20 Candida tropicalis, 16 Candida parapsilosis, 13 Candida glabrata, seven Candida kefyr, six Candida krusei, two of each Candida dubliniensis, Candida guilliermondi and one Candida lusitaniae. Compared to the results of the DNA sequence analysis, the accurate identification of the fully automated Phoenix™ system and the MALDI TOF-MS system was found as 92.3% and 97.8%, respectively. In addition to accuracy, costs and time of result periods of the three methods were also compared. Disregarding the cost of the device in the 3 methods, when the comparison was made of the cost per test and the time to results after pure production in SDA agar, the MALDI TOF-MS system was determined to have the lowest costs and provided results in the shortest time. As some of the Candida strains have antifungal resistance, identification of the strains must be a priority in respect of starting early treatment. The MALDI TOF-MS system has high performance in accurate identification, low costs and the system provides the results within minutes, thereby allowing immediate decision to be made for the antifungal treatment to be started. Thus, the morbidity, mortality and cost rates will be reduced. In conclusion, as the MALDI TOF-MS is a rapid, reliable and low cost per test system, it can be considered suitable for routine use in laboratories.

[Agents of otomycosis in Manisa region, Turkey, 1995-2011]. / Manisa Bölgesinde Otomikoz Etkenleri, 1995-2011.

Otomycosis, which is otitis externa caused by fungi, is common throughout the world especially in tropical and subtropical countries. However, the epidemiologic data about the etiologic agents of otomycosis in Turkey is limited. The aim of this retrospective study was to evaluate the agents of otomycosis in patients living at Manisa region (located at western Anatolia of Turkey). A total of 2279 cases [1465 male, 813 female; age range 1-87 (mean: 41.7) years] who were clinically prediagnosed as otomycosis at Celal Bayar University Hospital, between February 1995 and July 2011, were included in the study. External ear swab samples from patients with suspicion of otomycosis have been evaluated by routine mycological methods. Identification of mold-like fungi was based on colony morphology and microscopic examination of fungal structure, whereas germ tube test, growth characteristics on cornmeal-Tween 80 agar and API 20C AUX (bio-Mérieux, France) system were used for the identification of yeast-like fungi. Of the samples, 28% (638/2279) were found positive by direct microscopy and 24% (544/2279) by culture methods. Among culture-positive cases the isolation rates of mold-like and yeast-like fungi were 66% (359/544) and 34% (185/544), respectively. The number of distribution of the molds were as follows; Aspergillus niger (180), Aspergillus fumigatus (95), Aspergillus terreus (32), Aspergillus flavus (23), Aspergillus spp. (14), Penicillium spp. (13), Trichophyton spp. (T.rubrum 1, T.mentagrophytes 1); while this distribution was as follows for the yeasts; Candida tropicalis (97), Candida albicans (39), Candida parapsilosis (21), Candida glabrata (19), Candida kefyr (4), C.guilliermondii (2), Candida krusei (1), Geotrichum candidum (1) and Trichosporon capitatum (1). It was notable that 96% (344/359) of mold-like fungi were Aspergillus spp., and 99% (183/185) of yeast-like fungi were Candida spp. The results of this study indicated that the most frequent agents of otomycosis were non-dermatophyte species such as Aspergillus, followed by Candida. Dermatophytes were isolated in a small number of otomycosis cases. These data will provide support to the establishment of antifungal therapy guidelines for otomycosis.

Comparison of the efficacy of combination and monotherapy with caspofungin and liposomal amphotericin B against invasive candidiasis.

OBJECTIVE: To investigate if combination therapy with liposomal amphotericin B (LAmB), and caspofungin (CAS) is superior to monotherapies in an experimental model with azole-resistant Candida albicans. METHODS: This study was carried out between October 2006 and August 2007 in Celal Bayar University, Manisa, Turkey. A total of 144 mice were included in the study, and divided into 4 groups as: control (n=36), CAS treatment group n=36, LAmB treatment group (n=36), and combination therapy group (n=36). Treatment efficacy was assessed by determining survival, as well as, the decrease in tissue fungal densities. RESULTS: The fungal densities in tissues were significantly reduced, and the survival rates were prolonged with either CAS only, or LAmB only, or with combination therapy compared to those of controls (p<0.05). There was no significant difference between monotherapy groups. Decrease in tissue fungal densities were significant in CAS and LAmB (1mg/kg) combination group, compared to CAS (1mg/kg) and LAmB (1mg/kg) groups (p=0.004 for CAS, p=0.009 for LAmB). Survival rates were similar in both treatment groups. CONCLUSION: The combination treatment was superior with 1mg/kg of doses of LAmB and CAS in terms of reducing the tissue fungal burden. Although with combination therapy the survival rates prolonged in all subgroups, no significant difference between the combination and monotherapies could be shown. Additional studies with a large number of cases are warranted to investigate the superiority of combination therapy.

Use of DNA hybridization test for diagnosing bacterial vaginosis in women with symptoms suggestive of infection.

The purpose of this study was to evaluate a DNA hybridization test (Affirm VPIII) as an alternative to Gram stain for the rapid diagnosis of bacterial vaginosis in women with clinical signs of vaginal infection. Vaginal specimens were collected from 321 symptomatic women, and analyzed for bacterial vaginosis by both Gram stain using Nugent criteria and DNA hybridization test. Sensitivity, specificity, positive predictive value, and negative predictive value of the DNA hybridization test were determined using the Gram staining as the standard for diagnosis of bacterial vaginosis. Of the 321 patients, 115 (35.8%) were Gram positive for bacterial vaginosis and 126 (39.2%) were negative. 80 patients (25.0%) demonstrated intermediate Gram staining that was also considered negative. The Affirm system detected G. vaginalis in 107 (93.0%) of 115 vaginal specimens positive for bacterial vaginosis diagnosed by Gram stain. Compared to the Gram stain, DNA hybridization test had a sensitivity of 87.7% and a specificity of 96.0%. Positive and negative predictive values of the DNA hybridization test were 93.0% and 92.7%, respectively. In conclusion, Affirm VPIII hybridization test correlated well with Gram stain and may be used as a rapid diagnostic tool to exclude bacterial vaginosis in women with genital complaints.

Influence of oral intake of Saccharomyces boulardii on Escherichia coli in enteric flora.

Enteric flora constitutes 95% of the cells in the human body. It has been shown that the bacterial content of this flora is affected by diet and changes in nutrition. Considering that urinary tract infections (UTI) are mostly due to ascending infections from the gut flora, the importance of the elements of this flora and their characteristics becomes more evident. The aim of this study was to evaluate the influence of oral Saccharomyces boulardii (S. boulardii) intake on the number of Escherichia coli (E. coli) colonies in the colon. This study was carried out with 14 boys and 10 girls (total of 24 children) aged between 36 and 192 months (mean: 104.3+/-45.1 months). A commercial capsule or powder containing 5 billion colony-forming units (cfu) of S. boulardii was administered once a day for 5 days. The number of E. coli and yeast colonies was measured in the stool samples of the study group before and after the use of this drug. Before treatment, the mean number of E. coli colonies in g/ml stool was 384,625+/-445,744. This number decreased significantly to 6,283+/-20,283 after treatment (p=0.00). S. boulardii was not detected in stool before treatment and the number of colonies increased to 11,047+/-26,754 in g/ml stool. S. boulardii may be effective in reducing the number of E. coli colonies in stool. The influence of this finding on clinical practice such as prevention of UTI needs to be clarified by further studies.

The necessity of culture for the diagnosis of tinea pedis.

BACKGROUND: This study examined the consistency between the clinical diagnosis of tinea pedis and the results of direct fungal examination, prepared with 10% potassium hydroxide, and culture. METHODS: 2,427 patients clinically diagnosed with tinea pedis who presented to the mycology laboratory were reviewed retrospectively for the outcomes of direct fungal examination and culture. RESULTS: Direct examination was positive in 54.3% and culture was positive in 36.6% of the cases. The sensitivity and specificity of direct microscopy were 95.7% and 69.6%, respectively CONCLUSIONS: The clinical diagnosis of tinea pedis can be misleading, since it features lesions that can also be present in some other skin diseases and direct microscopy may be insufficient to confirm the diagnosis. Therefore, we suggest using culture for a definitive diagnosis.

Is skin disinfection with 10% povidone iodine sufficient to prevent epidural needle and catheter contamination?

BACKGROUND AND OBJECTIVES: Epidural space infection is a potential complication of epidural catheter placement. In this study, we investigated the incidence of epidural needle and catheter contamination after skin surface disinfection with 10% povidone-iodine (PI). METHODS: Sixty seven patients having surgery under epidural anesthesia were enrolled in this prospective study. After preparation with 10% PI, skin swab cultures were taken from the site of catheter insertion. Epidural needles were cultured immediately after epidural catheters were placed. Catheters were removed at 48 hours and 2 to 3 cm of the distal tips were cultured as well. RESULTS: Fifty-six skin swabs, 52 epidural needles, and 48 catheters were cultured. Although only 3.5% (2) colonization was observed on skin surface cultures, 34.6% (18) of the epidural needles and 45.8% (22) of the catheters were colonized. No systemic or local infection was observed. CONCLUSIONS: Our results suggest that despite skin surface disinfection with PI, there is still significant risk for contamination of needles and catheters during epidural catheterization.

Efficacy of azithromycin in a murine toxoplasmosis model, employing a Toxoplasma gondii strain from Turkey.

A murine toxoplasmosis model with Balb/C mice was used to investigate the therapeutic and prophylactic efficacy of azithromycin in a native strain of Toxoplasma gondii. Initially, seven groups--four studies and three controls--were established and 10(3) tachyzoites of this native strain of T. gondii were injected intraperitoneally to the mice in groups 1, 2, 3, 4 and 7. Azithromycin was given to groups 1-4 at different times of infection orally between 100 and 300 mg/kg/day for 10 days. Azithromycin was found to be effective at 200 mg/kg/day and above in the prophylaxis, at 250 mg/kg/day and above in the treatment of toxoplasmosis. These results suggest that azithromycin is effective in the prophylaxis and early infection of a highly virulent strain of T. gondii, and it doubled the survival time in the late infection. Azithromycin could be an alternative treatment regimen for human toxoplasmosis, if supported by further clinical investigations.

Giardiasis treatment in Turkish children with a single dose of ornidazole.

This study was designed to compare the treatment efficacy of single dose of ornidazole with 5 d treatments of ornidazole and metronidazole in children with giardiasis. 175 children, between 2 and 15 y old, whose stool samples were found to be positive for Giardia lamblia cysts and/or trophozoites by either saline-Lugol, formalin-ethyl acetate or trichrome staining, were enrolled in the study. Of these children, 105 were treated with a single dose of ornidazole: 35 with 30 mg/kg, 35 with 25 mg/kg and 35 with 20 mg/kg; 35 were treated with 25 mg/kg per day of ornidazole for 5 d in 2 doses and 35 children were treated with 20 mg/kg per day metronidazole for 7 d in 3 doses. All cases were examined on the 7th, 10th and 14th days after treatment by the same methods; clinical symptoms were also evaluated. Giardia lamblia was eradicated in 34 of 35 (97%), 34 of 35 (97%) and 33 of 35 (94%) patients treated with 30, 25 and 20 mg/kg single doses of ornidazole, respectively. Eradication was achieved in all 35 patients treated with 25 mg/kg per day ornidazole for 5 d and in 31 of 35 (89%) patients treated with metronidazole. There was no statistically significant difference among doses of ornidazole (p > 0.05); however, all ornidazole treatment regimens were significantly more effective than metronidazole treatment (p < 0.05). No important side-effects were detected in any patients and clinical symptoms disappeared in all. Single-dose ornidazole treatment could be considered as a proper and effective alternative method for the treatment of giardiasis in children.