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The aim of this study was to determine the Firmicutes/Bacteroidetes ratio in the gut microbiota and IL-1β, IL-6, IL-8, TLR2, TLR4 and TLR5 gene expression levels in the blood of adult type 2 diabetes (T2D) patients and compare it with that of adult nondiabetic healthy controls (HC).Between May 2016 and April 2017, 99 T2D patients and 99 HCs were enrolled in the study. Bacteroidetes and Firmicutes levels were assessed from stool sample DNA and IL-1β, IL-6, IL-8, TLR2, TLR4, and TLR5 gene expression levels assesed from blood sample RNA via qPCR from both T2D patients and healthy controls.The Firmicutes/Bacteroidetes ratio detected in the stool of type 2 diabetes patients was found to be higher with a statistically significant difference (p0.0001). Gene expression levels of IL-1β, IL-6, IL-8, TLR2, TLR4, and TLR5 were found to be upregulated.The highest upregulation was detected in IL-6 with 11 fold in T2D patients comparing with HCs. F/B ratio and gene expression levels were elevated in T2D patients. Firmicutes were positively correlated with studied gene expressions. A better understanding of the complex interaction between gut microbiota, environment, and diabetes will allow for more effective prevention and treatment strategies for T2D.

Akut gastroenterit (AGE), tüm dünyada yaygın olarak görülen enfeksiyon hastalıkları arasındadır. Campylobacter, Salmonella, Shigella türleri AGE’nin bakteriyel etkenleri arasında ilk sıralarda bulunmaktadır. Bu çalışmada, laboratuvarımıza gönderilen gaita örneklerinde kültür yöntemi ve moleküler yöntemle Campylobacter, Salmonella ve Shigella sıklığını saptamak ve her iki yöntemin kıyaslanması amaçlanmıştır. 2019-2020 yılları arasında Namık Kemal Üniversitesi Tıp Fakültesi Hastanesi’ne ishal yakınması ile başvuran, ayaktan erişkin ve çocuk hasta örnekleri dahil edilmiştir. Dışkı örnekleri Campylobacter, Salmonella ve Shigella türlerinin tespiti için konvansiyonel kültür yöntemleri ve polimeraz zincir reaksiyonu (PZR) yöntemi ile çalışılmıştır. Çalışmamızda 400 (%77 yetişkin, %23 çocuk) hastaya ait dışkı örneği değerlendirilmiştir. Örneklerin kültür ile değerlendirilmesinde 14 örnekte (%3.5) etken saptanmış; 10’u (%2.5) Campylobacter spp., 4’ü (%1) Salmonella spp. olarak tiplendirilmiş, Shigella spp. izole edilememiştir. Kültür yöntemi ile Campylobacter spp. ve Salmonella spp. sıklığı çocuklarda sırasıyla %6.5 ve %2.2, yetişkinlerde %1.3 ve %0.7 olarak saptanmıştır. Örneklerin PZR ile değerlendirilmesinde 38 örnekte (%9.6) etken saptanmış; 33’ünde (%8.3) Campylobacter spp., 5’inde (%1.3) Salmonella spp. varlığı tespit edilmiş, hiçbir örnekte Shigella spp. saptanamamıştır. Kültürde üreme saptanan 14 dışkı örneğinin tamamında PZR yönteminde de aynı etken mikroorganizma tespit edilmiştir. 24 örnekte ise kültürde üreme saptanmayıp sadece PZR’de Campylobacter spp. veya Salmonella spp. saptanmıştır. Gaita kültürü AGE tanısında altın standart yöntem olmasına rağmen araştırmamız sonucunda PZR’nin hem hızlı sonuç vermesi hem de saptama oranının yüksek olması nedeniyle kültüre avantajlı olduğu görülmüştür. Bu nedenle moleküler yöntemlerin rutin tanıda kullanılmasının faydalı olacağı kanaatindeyiz.

Objectives: “Nosocomial infections” or “healthcare-associated infections” are a significant public health problem around the world. This study aimed to assess the rate of laboratory-confirmed healthcare-associated infections, frequency of nosocomial pathogens, and the antimicrobial resistance patterns of bacterial isolates in a University Hospital. Methods: A retrospective evaluation of healthcare-associated infections in a University Hospital, between the years 2015 and 2019 in Tekirdag, Turkey. Results: During the 5 years, the incidence densities of healthcare-associated infections in intensive care units and clinics were 10.31 and 1.70/1000 patient-days, respectively. The rates of ventilator-associated pneumonia, central line–associated bloodstream infections, and catheter-associated urinary tract infections in intensive care units were 11.57, 4.02, and 1.99 per 1000 device-days, respectively. The most common healthcare-associated infections according to the primary sites were bloodstream infections (55.3%) and pneumonia (20.4%). 67.5% of the isolated microorganisms as nosocomial agents were Gram-negative bacteria, 24.9% of Gram-positive bacteria, and 7.6% of Candida. The most frequently isolated causative agents were Escherichia coli (16.7%) and Pseudomonas aeruginosa (15.7%). The rate of extended-spectrum beta-lactamase production among E. coli isolates was 51.1%. Carbapenem resistance was 29.8% among isolates of P. aeruginosa, 95.1% among isolates of Acinetobacter baumannii, and 18.2% among isolates of Klebsiella pneumoniae. Colistin resistance was 2.4% among isolates of A. baumannii. Vancomycin resistance was 5.3% among isolates of Enterococci. Conclusion: Our study results demonstrate that healthcare-associated infections are predominantly originated by intensive care units. The microorganisms isolated from intensive care units are highly resistant to many antimicrobial agents. The rising incidence of multidrug-resistant microorganisms indicates that more interventions are urgently needed to reduce healthcare-associated infections in our intensive care units.

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.

OBJECTIVE: ‘Nosocomial infections’ or ‘healthcare associated infections’ are a significant public health problem around the world. This study aimed to assess the rate of culture confirmed nosocomial infections (NIs), frequency of nosocomial pathogens and the antimicrobial resistance patterns of bacterial isolates in a University Hospital. METHODS: A retrospective evaluation of NIs in a tertiary hospital, between the years 2015 and 2019 in Tekirdag, Turkey. RESULTS : During the five years, the overall incidence rates (NI/100) and incidence densities (NI/1000 days of stay) of NIs were 2.04% (range 1.76-2.41/100) and 3.50/1000 patients-days (range 2.85-4.64/1000), respectively. 57.4 % of the infections were originated from the Intensive Care Units. The most common NIs according to the primary sites were bloodstream infections (55.3 %) and, pneumonia (20.4%). 67.5% of the isolated microorganisms as nosocomial agents were Gram negative bacteria, 24.9% of Gram positive bacteria and 7.6 % of candida. The most frequently isolated causative agents were Esherichia coli (16.7%) and Pseudomonas aeruginosa (15.7%). The rate of extended spectrum beta-lactamase production among E. coli isolates was 51.1%. Carbapenem resistance was 29.8% among isolates of Pseudomonas aeruginosa; 95.1% among isolates of Acinetobacter baumannii, 18.2% among isolates of Klebsiella pneumoniae. Colistin resistance was 2.4% among isolates of Acinetobacter baumannii. Vancomycin resistance was 5.3% among isolates of Enterococci.CONCLUSION: Our study results demonstrates the microorganisms of isolated from Intensive Care Units demonstrates high level resistance to many antimicrobial agents. The rising in incidence of multidrug-resistant microorganisms indicate that more interventions are urgently needed to reduce NIs in our ICUs.

Bu calismada hastanemizde izole edilen Candida turlerinin iki farkli yontemle antifungal duyarliliklarinin incelenmesi amaclanmistir. Hastanemizde cesitli departmanlardan gelen klinik orneklerden toplam 200 Candida turu izole edildi. Bu turler geleneksel yontem (germ tup olusturma, corn-meal tween 80 besiyerinde mikroskobik gorunum) ve ticari kit API 20C (Biomerieux, France) ile tanimlandi. Bu turlerin flukonazol (FLU), vorikonazol (VORI), amfoterisin B (AMB) antifungal minimum inhibisyon konsantrasyonu (MIK) degerleri Clinical and Laboratory Standards Institute sivi mikrodilusyon ve disk-difuzyon (DD) yontemine gore degerlendirildi. Izole edilen 200 Candida turu; 100 C. albicans, 25 C. glabrata, 7 C. krusei, 43 C. parapsilosis ve 25 C. tropicalis olarak tanimlandi. Butun Candida izolatlari VORI ‘ye karsi duyarli bulundu. Yuksek direnc (%22,5) orani referans mikrodilusyon yontemi ile AMB antifungaline karsi tespit edilidi. Iki yontem arasinda en iyi kategorik oran sirasi ile VORI (%99), FLU (%84,9) ve AMB (%72,5) olarak hesaplandi.Vorikonazol, flukonazole direncli izolatlarda bile in vitro olarak flukonazolden daha iyi aktivite gostermistir. Ek olarak disk difuzyon yontemi VORI icin alternatif antifungal duyarlilik yontemi olarak degerlendirilebilir.

IntroductionBurkholderia cepacia complex (Bcc) leads to severe nosocomial infections particularly in the patients who have intravascular catheters and cystic fibrosis. The present study aims at investigation of Bcc outbreaks in non-cystic fibrosis patients.Material and MethodsA total of 6 patients who were hospitalized at General Pediatrics Department were included in the study. Blood cultures which yielded positive signals were incubated at 5% blood sheep agar, chocolate agar and Eosin Methylene Blue agar. All fields which could be the source of the infection at the clinic were examined. Isolates confirmation with Pulsed-Field Gel Electrophoresis (PFGE) tests were performed.ResultsThe first patient aged 14.5 years was hospitalized due to left renal agenesis, urinary tract infection and renal failure. Bcc growing was detected in blood culture which was obtained due to high fever at the 3rd day of hospitalization. New patient hospitalizations were stopped due to Bcc growing in blood cultures which was obtained due to high fever in the remaining five patients. No growing was detected in samples obtained from the clinic and the patient rooms. PFGE patterns were similar in all clinical isolates of Bcc indicating that the outbreak had originated from the same origin.ConclusionsBcc infection should always be kept in mind in nosocomial outbreaks due to multi-drug resistance and the need for hospitalization at intensive care unit. Control measures should be taken for prevention of nosocomial infections and required investigations should be done for detection of the source of the infection.

Invasive fungal infections are a leading cause of morbidity and mortality in immunocompromised patients, especially in cases requiring a prolonged stay in the intensive care unit. A total of 99 yeast strains were isolated from 42 postmortem cases. In this study, virulence factors and antifungal susceptibility of these species were evaluated. The isolates were identified as Candida albicans (54), C. tropicalis (15), C. glabrata (12), C. parapsilosis (6), C. lipolytica (3), C. utilis (3), C. krusei (2), C. kefyr (1), and Cryptococcus neoformans (3). The most commonly isolated species was C. albicans, and no resistant species were determined. Despite the equal number of specimens, no secretion of significant virulence factors was associated with the postmortem specimen in the Candida species. Postmortem fungal investigations in forensic autopsies are useful in explaining cause of death in such cases, also may lead to protocols for the treatment of fungal infections and contribute to fungal pathogenesis and epidemiological data.

The aim of this study was to determine and compare the levels of both Bacteroidetes and Firmicutes in the gut microbiota and TLR2/TLR4 gene expression in the blood of patients with type 1 diabetes mellitus (T1DM) and healthy individuals. These results may serve as a preliminary assessment to guide future research.Between January and October 2014, stool and blood samples were collected from 53 adult T1DM patients and 53 age- and gender-matched healthy individuals. Bacteroidetes and Firmicutes levels were assessed from stool sample DNA and TLR2 and TLR4 expression levels were analyzed via qPCR using RNA from EDTA blood samples from both patients and healthy controls.The amounts of Bacteroidetes and Firmicutes were statistically significantly higher and lower, respectively, in the T1DM group than in the healthy control group (p 0.001 and p 0.001, respectively). In addition, the Firmicutes/Bacteroidetes ratios in patients with T1DM were significantly lower than in healthy controls. The TLR4 and TLR2 gene expression levels in T1DM patients were significantly upregulated and downregulated, respectively, compared to those in the control group.Our data are the first to show a relationship between T1DM and gut microbiota in our country. In addition, our results provide information about the connections between T1DM, gut microbiota, and TLR2 and TLR4 expression. We believe that Bacteroidetes and Firmicutes in the gut microbiota may play a role in the autoimmune process of T1DM and that these findings should be further investigated in the future.

Autism spectrum disorder (ASD) is a general term for a group of complex neurodevelopmental disorders of brain development that limits a person’s ability to function normally. Etiology has not been clearly defined up to date. However, gut microbiota and the bidirectional communication between the gastrointestinal tract and brain, the so-called microbiota–gut–brain axis, are hypothesized, which may be involved in the etiology of several mental disorders. Recent reports suggest that Candida, particularly Candida albicans, growth in intestines may cause lower absorption of carbohydrates and minerals and higher toxin levels which are thought to contribute autistic behaviors. The aim of this study was to identify the 3-year deposited yeasts isolated from stool samples of children with diagnosed or suspected ASD and to determine in vitro activity of nystatin and fluconazole against these isolates using Clinical Laboratory Standards Institute M27-A3 guidelines. A 17-year retrospective assessment was also done using our laboratory records. Among the species identified, intrinsically fluconazole-resistent Candida krusei (19.8 %) and Candida glabrata (14.8 %) with elevated MICs were remarkable. Overall, C. albicans (57.4 %) was the most commonly isolated species in 17 years. The species identification and/or antifungal susceptibility tests have to be performed using the strain isolated from stool sample, to select the appropriate antifungal agent, if antimycotic therapy is needed.

Bacillus species can be found in air, water, soil, feces, and particularly in the normal flora of patients with a prolonged length of hospitalization. Therefore, isolation of these microorganisms from clinical samples is a common condition and usually regarded as contaminants. Previously, Bacillus cereus has been rarely associated with infections, but in the last few years, it has been reported with increasing frequency as the bacterial pathogen in patients with intravenous drug abuse, trauma, immunodeficiency syndrome, immunosuppressed patients with a history of underlying malignancy and granulocytopenia. In this study, we present a 3-months-old baby girl who was presented to a private health center with a complaint of not sucking well enough. After clinical examination she was diagnosed with neuroblastoma stage 4 S, metastasis was detected in the liver and bone marrow and surrenalectomy and 14 cures of chemotherapy were applied to the patient. Patient was discharged from the hospital with a permanent tunneled catheter, but she returned to the pediatric emergency department with high fever. After laboratory tests she was diagnosed with febrile neutropenia and ceftazidime and amikacin treatment was started. Ceftazidime treatment was stopped after the isolation of B. cereus from peripheral and catheter blood cultures of the patient and teicoplanin treatment was started. Permanent tunneled catheter was removed under anesthesia, and this suggested the case as a catheter-originated bacteremia after reducing fever of the patient. As a result, we suggest that B. cereus isolation in blood cultures could not everytime mean contamination, and B. cereus may cause catheter-related infections in immunosuppressed patients. (JAREM 2015; 5: 75-7)

Background: C. krusei is an opportunistic fungal pathogen that known of its intrinsic fluconazole resistance and its frequency is increasing especially among hematology patients. The increase in the frequency of high mortality fungal infections have accelerated the efforts of new drug development with broad spectrum, low toxicity and the studies including their combination. However, there is no standardized method to evaluate the activity of drug combinations. Aims: To evaluate the activity of caspofungin (CAS) with voriconazole (VOR) and amphotericin B (AMB) alone and in combination and the utility of Etest and disk diffusion methods for antifungal combinations. Methodology: The minimum inhibitory concentrations of VOR, CAS and AMB against 30 clinical C. krusei isolates were determined by using Etest, disk diffusion and reference broth microdilution methods. Combinations of CAS with VOR and CAS with AMB were evaluated using disk diffusion (three different ways) and Etest (two different ways) methods. Original Research Article British Microbiology Research Journal, 4(5): 497-508, 2014 498 Results: All isolates tested were susceptible to VOR and CAS in vitro by all three methods. Categorical agreements of Etest and disk diffusion methods with reference microdilutiontest were 100% for CAS and VOR (for each method), 86.7% and 50% for AMB, respectively. In the all ways of both combination methods, we did not observe distinctly antagonistic or synergic interaction. Conclusion: Etest and disk diffusion could be easy, convenient, and nontime-consuming alternative methods to evaluate the antifungal combinations. The combinations of CAS with VOR and AMB exhibited promising results because of an apparent antagonistic interaction was not detected in this study.

Invasive aspergillosis usually affects the lungs, and osteomyelitis due to Aspergillus spp. is a rare manifestation of invasive aspergillosis. Generally, there are underlying immunosuppressive conditions such as prolonged granulocytopenia, immunosuppressive therapy, and chronic granulomatous disease. Although amphotericin B is one of the most preferred agents for invasive aspergillosis management, its poor bone penetration and renal toxicity limit its use for the treatment of osteomyelitis, especially in pediatric ages. Case Report / Olgu Sunumu

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).

Contains fulltext : 171933.pdf (Publisher’s version ) (Closed access) Investigations of both virulence factors and antifungal susceptibility profiles are crucial for understanding the pathogenesis and prognosis of ophthalmic mycoses. In this study, we investigated the in vitro antifungal susceptibility of amphotericin B (AMB), voriconazole (VRC), and natamycin (NAT) against a set of 50 fungal isolates obtained from patients with ocular mycoses using the Clinical and Laboratory Standards Institute broth microdilution method. In addition, putative virulence factor, such as secretory phospholipases and proteinases, and biofilm formation activity were analyzed. The geometric means (GMs) of the minimum inhibitory concentrations (MICs) of the antifungals across all isolates were the following (in increasing order): VRC (0.70 mug/mL), AMB (0.81 mug/mL), and NAT (1.05 mug/mL). The highest activity against 14 Aspergillus strains was exhibited by VRC (GM MIC: 0.10 mug/mL), followed by AMB and NAT (GM MICs: 0.21 and 0.27 mug/mL), respectively. However, for 12 Fusarium spp., the GM MIC of VRC (2.66) was higher than those of NAT and AMB (GM MICs 1.3 and 0.8 mug/mL, respectively). Proteinase and phospholipase activity were observed in 30 % and 42 % of the isolates, respectively, whereas only 8 % of the isolates were able to produce biofilms. Phospholipase activity was observed in all Fusarium isolates, but not in any of the Aspergillus isolates. In contrast, biofilm-forming capability was detected in 25 % of the Fusarium isolates, but none of the Aspergillus isolates. The differences in the MICs of AMB, VRC, and NAT, biofilm-forming ability and proteinase and phospholipase activities among the isolates were not significant (p > 0.05). Overall, our study suggests no significant correlation between the antifungal susceptibility profiles and virulence attributes of ocular fungal isolates.

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.

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.

Anaerobic bacteria play an important role in eye infections; however, there is limited epidemiologic data based on the the role of these bacteria in the etiology of keratitis and endophthalmitis. The aim of this re- search is to determine the prevalence of anaerobic bacteria in perforated corneal ulcers of patients with keratitis and endophthalmitis and to evaluate their antimicrobial susceptibilities.Corneal scrapings were taken by the ophthalmologist using sterile needles. For the isolation of anaerobic bacteria, samples were inoculated on specific media and were incubated under anaerobic conditions obtained with Anaero-Gen (OxoidMitsubishi Gas Company) in anaerobic jars (Oxoid USA, Inc. Columbia, MD, USA). The molecular identification of anaerobic bacteria was performed by multiplex PCR and the susceptibilities of an- aerobic bacteria to penicillin, chloramphenicol, and clindamycin were determined with the E test (bioMerieux).51 strains of anaerobic bacteria belonging to four different genuses were detected by multiplex PCR and only 46 strains were isolated by culture. All of them were found susceptible to chloramphenicol whereas penicillin resistance was found in 13.3% of P.anaerobius strains, clindamycin resistance was found in 34.8% of P.acnes and 13.3% of P. anaerobius strains. Additionnaly, one strain of P. granulosum was found resistant to clindamycin, one strain of B. fragilis and one strain of P.melaninogenica were found resistant to penicillin and clindamycin.Routine analyses of anaerobes in perforated corneal ulcers is inevitable and usage of appropriate molecular methods, for the detection of bacteria responsible from severe infections which might not be deter- mined by cultivation, may serve for the early decision of the appropriate treatment. Taking into account the in- creasing antimicrobial resistance of anaerobic bacteria, alternative eye specific antibiotics effective against anaer- obes are needed to achieve a successful treatment.

A 5-year-old boy with acute lymphoblastic leukemia and probable pulmonary aspergillosis developed a hard, protuberant, white-yellow and aggressive elongated lesion on the left side of his tongue during a febrile agranulocytopenic episode. Despite the use of broad-spectrum antibiotics and other supportive therapies, the lesion increased to about 2x4 cm in size within two days and became grey-black with an erythemateous, irregular line. Partial excision of the tongue was performed and isolates recovered from the tongue biopsy specimen were identified as Aspergillus flavus. An increase in the systemic dose (7 mg/kg/day) and local intraoral delivery of liposomal amphotericin B was successful in treating the patient and resulted in improved clinical and laboratory findings. Herein, we document the observation of tongue aspergillosis in a leukemic child with probable pulmonary aspergillosis receiving liposomal amphotericin B therapy and the successful treatment of tongue aspergillosis with an increased dose (7mg/kg) of liposomal amphotericin B. To the best of our knowledge, this is the youngest patient with documented intraoral aspergillosis and only the second case of tongue aspergillosis caused by Aspergillus flavus.

The aim of this study was to investigate the susceptibility of the different phenotypes of Candida albicans strains isolated from clinical specimens to three antifungal agents, fluconazole, itraconazole and voriconazole. Totally 215 specimens were collected from oropharyngeal, gastrointestinal and urogenital tracts of non-neutropenic patients who had received no previous prophylactic treatment. Each of the 215 C. albicans strains recovered was found to express one of the six phenotypes: smooth 73%, fuzzy 10.7%, irregular 2.3%, star 2.8%, ring 6% or stipple 5.1%. The mean MICs for the six phenotypes of C. albicans strains ranged between 0.25 microg/ml and 64 microg/ml for fluconazole, 0.03 microg/ml and 1 microg/ml for itraconazole and 0.03 microg/ml and 0.5 microg/ml for voriconazole. The mean minimum inhibitory concentration (MIC) of fluconazole was consistently higher for C. albicans strains expressing the stipple phenotype. The antifungal susceptibility of the phenotypic switching requires attention, especially in patients who are clinically unresponsive to fluconazole chemotherapy or in cases of serious C. albicans infections of immunocompromised hosts.

Summary The aim of our study was to determine the prevalence of tinea capitis in Middle Anatolia. Between February 2000 and April 2000, a school survey of 4760 primary school children was carried out and 34 cases of tinea capitis were clinically diagnosed. Sixteen (0.3%) of them were diagnosed both clinically and using culture. Eighteen cases (52.9%) were diagnosed microscopically whereas the remaining 16 cases (47.0%) had a clinical diagnose. We also isolated and identified the following species of fungi causing the infection: Trichophyton verrucosum (five cases), T. rubrum (three), T. mentagrophytes var. mentagrophytes (three), Microsporum canis (three) and T. tonsurans (two). We detected a higher rate of infection than that of the country's average. Zusammenfassung Ziel unserer Studie war die Bestimmung der Pravalenz von Tinea capitis in Mittelanatolien. In einer Schule mit 4760 Grundschulschulern wurden 34 Falle von Tinea capitis klinisch diagnostiziert. 16 (0.3%) Tinea capitis-Infektionen wurden klinisch und mikrobiologisch diagnostiziert. Mikroskopisch wurden 18 positive Falle (52.9%) festgestellt, 16 Falle wurden klinisch diagnostiziert. Trichopyton verrucosum (5 Falle), T. rubrum (3), T. mentagrophytes var. mentagrophytes (3), Microsporum canis (3) und T. tonsurans (2). Die von uns bestimmte Pravalenz lag hoher als die des Landesdurchschnitts.

Background Porphyromonas gingivalis, a major periodontal pathogen, is gaining increasing attention for its possible association with atherosclerosis. Its fimbriae are classified into six genotypes (Types I-V, Ib) based on the diversity of the fim A genes encoding the fimbrial subunits. In this study, fim A genotype's distribution of P. gingivalis was analyzed in atherosclerotic plaque specimens. Methods A total of 50 atherosclerotic plaque specimens and 50 non-atherosclerotic, post stenotic aneurysm specimens were collected from patients undergoing cardiovascular surgery. Bacterial DNA was also extracted from each specimen, as real-time PCR was carried out with P. gingivalis-specific primer sets. The positive specimens of P. gingivalis were further analyzed to discriminate the fim A genotype using real-time and nested PCR methods. Results P. gingivalis was detected only in one atherosclerotic plaque; however, the genotype was nontypable in this specimen. Conclusions We state that it is not easy to show a significant relationship between P. gingivalis, its fim A genotype, and atherosclerosis. We suggest that new extended studies based especially upon the quantitave determination of P. gingivalis and its genotype distribution on atherosclerotic specimens are needed to show an evident relationship between atherosclerosis and P. gingivalis.

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.

We report a case of fungaemia resulting from Candida norvegensis in a patient with acute non-lymphoblastic leukaemia-M4 from Turkey. Candida norvegensis was isolated from two different peripheral blood samples that were taken at 2-day intervals. Despite treatment with liposomal amphotericin B, the patient died of multi-organ system failure.

In this study, some 1-[2-(substituted pyrrol-1-yl)ethyl]-2-methyl-5- nitroimidazole derivatives were synthesized by reacting 2-(2-methyl-5-nitro- 1H-imidazolyl)ethylamine dihydrochloride, which was prepared using metronidazole, with some 1,4-dicarbonyl compounds. The structure elucidation of the compounds was performed by IR, 1H-NMR and mass spectroscopic data and elemental analysis results. Antimicrobial activities of the compounds were examined and notable activity was obtained.

To evaluate the efficacy of caspofungin in an experimental rabbit model of Fusarium keratitis and to compare it with amphotericin B.Eighteen New Zealand white rabbits were randomly divided into 2 treatment groups and 1 control group. One cornea of each rabbit was inoculated with Fusarium solani spores. The first group received topical amphotericin B 0.15%, the second group received topical caspofungin 1%, and the control group received topical balanced salt solution hourly for 2 days and then 4 times daily for 3 additional days. Treatment effects were evaluated by clinical assessment at days 3 and 5 and by fungal culture after 5 days of treatment.In the treatment groups, progression of keratitis was inhibited, and cultures were sterile at the end of the study. In the control group, keratitis progressed, and cultures were positive for F. solani.Topical caspofungin is effective in Fusarium keratitis, and clinical efficacy studies seem justified.

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.

Background Anaerobic bacteria play an important role in eye infections; however, there is limited epidemiologic data based on the the role of these bacteria in the etiology of keratitis and endophthalmitis. The aim of this re- search is to determine the prevalence of anaerobic bacteria in perforated corneal ulcers of patients with keratitis and endophthalmitis and to evaluate their antimicrobial susceptibilities. Methods Corneal scrapings were taken by the ophthalmologist using sterile needles. For the isolation of anaerobic bacteria, samples were inoculated on specific media and were incubated under anaerobic conditions obtained with Anaero-Gen (Oxoid & Mitsubishi Gas Company) in anaerobic jars (Oxoid USA, Inc. Columbia, MD, USA). The molecular identification of anaerobic bacteria was performed by multiplex PCR and the susceptibilities of an- aerobic bacteria to penicillin, chloramphenicol, and clindamycin were determined with the E test (bioMerieux). Results 51 strains of anaerobic bacteria belonging to four different genuses were detected by multiplex PCR and only 46 strains were isolated by culture. All of them were found susceptible to chloramphenicol whereas penicillin resistance was found in 13.3% of P.anaerobius strains, clindamycin resistance was found in 34.8% of P.acnes and 13.3% of P. anaerobius strains. Additionnaly, one strain of P. granulosum was found resistant to clindamycin, one strain of B. fragilis and one strain of P.melaninogenica were found resistant to penicillin and clindamycin. Conclusions Routine analyses of anaerobes in perforated corneal ulcers is inevitable and usage of appropriate molecular methods, for the detection of bacteria responsible from severe infections which might not be deter- mined by cultivation, may serve for the early decision of the appropriate treatment. Taking into account the in- creasing antimicrobial resistance of anaerobic bacteria, alternative eye specific antibiotics effective against anaer- obes are needed to achieve a successful treatment.

Cladophialophora bantiana is a phaeoid fungus that only rarely has been isolated from sources other than the human brain. It has a particular tropism for the central nervous system (CNS). We have integrated and updated large-scale data related to several aspects of C. Bantiana and reviewed all the available reports on its cerebral infections, focusing on their geographical distribution, infection routes, immune status of infected individuals, type and location of infections, clinical manifestations and treatment and outcome, briefly looking over the spectrum of other disease entities associated with C. bantiana, that is, extra-cerebral and animal infections and on the environmental sources of this fungus. Among the agents of phaeohyphomycosis, a term used to describe an infection caused by a dark pigmented fungus, C. bantiana has some significant specific features. A total of 120 case reports were identified with a significantly higher percentage of healthy subjects than immune-debilitated patients (58.3% vs. 41.7%). Infections due to C. bantiana occur worldwide. The main clinical manifestations are brain abscess (97.5%), coinfection of brain tissue and meninges (14.2%) and meningitis alone (2.5%). Among immunocompetent patients, cerebral infection occurred in the absence of pulmonary lesions. The mortality rate is 65.0% regardless of the patient's immune status. The therapeutic options used include surgery or antifungals alone, and the combination of both, in most cases the fatal outcome being rapid after admission. Since the fungus is a true pathogen, laboratory workers should be made aware that BioSafety Level-3 precautions might be necessary.

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.