European candidaemia is characterised by notable differential epidemiology and susceptibility pattern: Results from the ECMM Candida III study.

The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25-33%) while Italy and Turkey had the highest C. parapsilosis proportions (24-26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis, 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks-alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence.

Incidence and Risk Factors for Antibiotic-associated Diarrhea Among Hospitalized Children.

BACKGROUND: We aimed to evaluate the incidence, clinical findings, and risk factors of antibiotic-associated diarrhea (AAD) in hospitalized children without known comorbid diseases. METHODS: All hospitalized children during the 1-year period that fulfilled the inclusion criteria were included in this study (n = 358). AAD was defined as; ≥2 loose or watery stools per day for a minimum of 24 hours during antibiotic treatment caused by Clostridioides difficile or negative stool tests for identifiable infectious agents. RESULTS: During hospitalization, diarrhea developed in 32 (8.93%) of the 358 patients. C. difficile toxin B was positive for 1 case. No infectious agents were detected in 21 patients. Overall, AAD was observed in 22 patients (6.14%, 95% CI: 4.09-9.13). Male sex ( P = 0.027, OR: 3.36), age between 1 month and <3 years ( P = 0.01, OR: 4.23), ibuprofen use ( P = 0.044, OR: 2.63) and late administration of antibiotics ( P = 0.001, OR: 9.5) were associated with the development of AAD. CONCLUSIONS: The incidence of AAD is low among hospitalized children without comorbid diseases, and most diarrheal episodes are mild and self-limiting. The use of probiotics in this patient group may be limited to certain specific situations.

Neutralization of Wild-Type and Alpha SARS-CoV-2 Variant by CoronaVac® Vaccine and Natural Infection- Induced Antibodies.

One of the immune responses desired to be achieved by SARS-CoV-2 vaccination is to create neutralizing antibodies (nAbs), thus preventing the development and spread of infection. The aim of this study was to investigate the seropositivity rate, anti-spike antibody levels, and neutralizing capacity of these antibodies against wild type (WT) and alpha variants in serum samples of individuals who had been naturally infected or vaccinated with CoronaVac®. Total anti-spike antibody levels were determined in all samples. Neutralization assays were performed by the reduction of the cytopathic effect in Vero-E6 cells with infectious WT and alpha SARS-CoV-2 variants. Although both naturally infected and vaccinated individuals were all seropositive for antispike antibodies, 84.8% of the vaccinated group, and 89.3% of the naturally infected group had detectable nAbs. The nAbs titers were significantly higher in the naturally infected group for both WT and alfa variant of the virus as compared to the vaccinated individuals. In this study, it was observed that all individuals became seropositive six weeks after exposure to the vaccine or the virus. Moreover, naturally infected individuals had higher levels of nAbs than those vaccinated. The presence of nAbs against the alpha variant in both naturally infected and vaccinated individuals suggests that these antibodies may also be protective against infections, which may be caused by other variants, such as delta and omicron.

Serotype distribution of Streptococcus pneumoniae and pneumococcal vaccine coverage in adults in Turkey between 2015 and 2018.

OBJECTIVE: To evaluate the serotype distribution and antibiotic resistance in pneumococcal infections in adults and to provide a perspective regarding serotype coverage of both current and future pneumococcal vaccines. PATIENTS AND METHODS: This passive surveillance study was conducted with the Streptococcus pneumoniae strains isolated from the specimens of patients with pneumonia (materials isolated from bronchoalveolar lavage), bacteraemia, meningitis, pleuritis and peritonitis between 2015 and 2018. Serogrouping and serotyping were performed by latex particle agglutination and by conventional Quellung reaction using commercial type-specific antisera, respectively. The strains were analysed for penicillin, cefotaxime, erythromycin and moxifloxacin susceptibilities by E-test. RESULTS: In the whole study group (410 samples from adults aged ≥18 years), the most frequent serotypes were 3 (14.1%), 19 F (12%) and 1 (9.3%). The vaccine coverage for PCV13, PCV15, PCV20 and PPV23 was 63.9%, 66.6%, 74.1% and 75.9%, respectively, in all isolates. Penicillin non-susceptibility in invasive pneumococcal disease (IPD) was 70.8% and 57.1% in the patients aged <65 and ≥65 years, respectively. About 21.1% and 4.3% of the patients with and without IPD had cefotaxime resistance. Non-susceptibility to erythromycin and moxifloxacin was 38.2% and 1.2%, respectively. CONCLUSIONS: The results revealed that novel PCV vaccines may provide improved coverage as compared with the currently available vaccine, PCV13. The significant antibiotic resistance rates imply the need to extend the serotype coverage of the vaccines. Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution and incidence changes of IPD cases in the population and to inform policy makers to make necessary improvements in the national immunization programmes.Key messagesThis multicentre study demonstrated the most recent serotype distribution and antibiotic resistance in adult population in Turkey.Shifting from PCV13 to novel conjugated vaccines will significantly increase the coverage.Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution changes and the incidence of cases with invasive pneumococcal disease in the population.

Identification and antifungal susceptibility of Saprochaete clavata from invasive infections in Turkey.

Saprochaete clavata is an emerging opportunistic pathogen, that causes life-threatening infections, but there are limited evidence and information about the evaluation of in vitro antifungal susceptibility test results. The aim of this study was to determine S. clavata isolates from clinical specimens and to investigate their in vitro antifungal susceptibility. S. clavata was identified by API ID20C AUX (BioMérieux, Brussels, Belgium), MALDI TOF (Bruker Daltonik, Germany), and ITS gene region sequencing. In vitro susceptibility tests were performed using Sensititre YeastOne (TREK Diagnostic System, East Grinstead, UK). During the study period, 4,736 fungi were isolated from various clinical samples and, S. clavata was identified in eight patients with underlying diseases namely, pancreatic neoplasma, acute myeloid leukaemie, follicular lymphoma, cholelithiasis. Anidulafungin and micafungin minimum inhibitory concentration values were 1-2 and 1-4 mg/L, respectively, while those of the azole group antifungals were much lower. This is the first study in Turkey reporting isolation, identification and antifungal susceptibilities of S. clavata from clinical specimens. Higher MIC values seen in some isolates suggest that continuous monitoring of sensitivity rates and observation of regional differences will thus be useful guides in determining infection control and antifungal use policies.

[Investigation of Plasmid Mediated mcr Colistin Resistance Gene in Clinical Enterobacterales Isolates]. / Klinik Enterobacterales Izolatlarinda Plazmit Aracili mcr Kolistin Direnç Geninin Arastirilmasi.

Carbapenems are used in the treatment of infections caused by multidrug-resistant bacteria and colistin (polymyxin E) is used as the last choice of antimicrobial agent in those resistant to carbapenems. The worldwide and increased use of colistin, which causes cell death by disrupting the permeability of the cytoplasmic membrane of gram-negative bacteria, raised the problem of resistance. The transferable colistin resistance enzyme mcr, is a phosphoethanolamine transferase that adds phosphoethanolamine to lipid A and modifies lipopolysaccharides, leading to polymyxin resistance. The aim of this study was to investigate some of the most prevalent plasmid mediated colistin and carbapenemase resistance genes in colistin resistant Enterobacterales isolates. Enterobacterales isolates which were isolated in the samples of patients treated in the clinical units between October 2016 and September 2018 in the Karadeniz Technical University Faculty of Medicine Farabi Hospital Medical Microbiology Laboratory were included in the study. In addition to conventional methods, isolates were identified to the species level by MALDI-TOF MS (Bruker Daltonics, Germany). The antibiotic susceptibilities of Enterobacterales isolates were studied by an automated microbiology system (Phoenix, Becton Dickinson, USA) and evaluated according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. In isolates that are resistant to colistin, and the isolates that are found to be sensitive but should be included in the patient report of the colistin susceptibility test, colistin susceptibility tests were repeated with liquid microdilution method in accordance with EUCAST standards. The presence of extended spectrum beta-lactamase (ESBL), AmpC beta-lactamase and carbapenemase were determined by phenotypic methods according to EUCAST recommendations in colistin resistant Enterobacterales isolates. Furthermore, resistance genes of mcr-1-5, blaOXA-48, blaKPC, blaNDM, blaVIM, blaIMP were detected by polymerase chain reaction (PCR) method, followed by nucleotide sequence analysis of the amplified products. In our study, 14657 Enterobacterales isolates belonging to 7535 patients treated in different clinical units were examined retrospectively. Escherichia coli 61.2% (n= 8968), Klebsiella pneumoniae 22.7% (n= 3334) and Enterobacter cloacae 6.9% (n= 1005) were the most prevalent isolates. Carbapenem resistance was detected in 894 isolates, and 5.8% (n= 412) of 7135 isolates isolated between October 2016 and September 2017; 6.4% (n= 482) of 7522 isolates between October 2017 and September 2018 were found to be resistant. Considering all isolates, colistin resistant isolates were 65 (0.9%) between October 2016 and September 2017 and 97 (1.3%) between October 2017 and September 2018. By including only the first isolates in the study for the same agent growths in different samples of the same patient, 46 colistin resistant isolates were selected. Six isolates which could not be cultivated from stock cultures were excluded from the study material. Thirteen (32.5%) of the 40 colistin resistant Enterobacterales isolates were isolated in 2017 and 27 (67.5%) were isolated in 2018. ESBL was detected in 22, AmpC beta-lactamase was detected in 6, carbapenem resistance was detected in 15 of them by phenotypic methods. As a result of PCR analysis, mcr-1 gene detected in 2 isolates, blaOXA-48 in 2 isolates, blaVIM in 1 isolate, blaKPC and blaOXA-48 in 1 isolate, blaNDM and blaOXA-48 in 5 isolates. These results were confirmed by sequencing of the PCR products. The mcr-1 genes were found in E.coli isolates grown in urine culture samples of 2 women over 65 years of age treated in our hospital. Among the antibiotics tested, only ampicillin resistance was observed in 1 of the patients, whereas ampicillin, amoxicillin-clavulanate and ciprofloxacin resistance were detected in the other. In conclusion, as far as we can reach in the literature our publication is the first study showing the presence of mcr-1 gene in clinical samples in our country and confirmed by DNA sequence analysis. The detection of mcr gene in isolates without multidrug resistance showed once again the importance of colistin susceptibility testing in the laboratories. In addition, the presence of isolates containing more than one resistance genes in our study, suggests that the spread of carbapenem and colistin resistance may be faster than expected.

Serotype distribution of Streptococcus pneumonia in children with invasive disease in Turkey: 2015-2018.

Objectives: To determine the serotype distribution of pneumococcus causing invasive pneumococcal disease (meningitidis, bacteremia and empyema) in children in Turkey, and to observe potential changes in this distribution in time to guide effective vaccine strategies. Methods: We surveyed S. pneumoniae with conventional bacteriological techniques and with real-time polymerase chain reaction (RT-PCR) in samples of cerebrospinal fluid (CSF), blood and pleural fluid. S. pneumoniae strains were isolated from 33 different hospitals in Turkey, which are giving health services to approximately 60% of the Turkish population. Results: A total of 167 cases were diagnosed with invasive pneumococcal disease between 2015 and 2018. We diagnosed 52 (31.1%) patients with meningitis, 104 (62.2%) patients with bacteremia, and 11 (6.6%) patients with empyema. Thirty-three percent of them were less than 2 years old and 56% less than 5 years old. Overall PCV13 serotypes accounted for 56.2% (94/167). The most common serotypes were 19 F (11.9%), 1 (10.7%) and 3 (10.1%). Conclusions: Besides the increasing frequency of non-vaccine serotypes, vaccine serotypes continue to be a problem for Turkey despite routine and high-rate vaccination with PCV13 and significant reduction reported for the incidence of IPD in young children. Since new candidate pneumococcal conjugate vaccines with more serotype antigens are being developed, continuing IPD surveillance is a significant source of information for decision-making processes on pneumococcal vaccination.

[Comparison of mRNA Levels of Stimulator of Interferon Genes (STING) in Individuals with Natural Immunity to Hepatitis B Virus (HBV), and in those with Chronic Hepatitis B Infection and without HBV]. / Hepatit B Virüsüne (HBV) Karsi Dogal Bagisik, Kronik HBV Enfeksiyonu Olan ve HBV ile Karsilasmamis Bireylerdeki "Stimulator of Interferon Genes (STING)" mRNA Düzeylerinin Karsilastirilmasi.

It has been estimated that currently 350-400 million people have been chronically infected with the hepatitis B virus (HBV) worldwide and approximately one million people die each year due to HBV related diseases. It has been suggested that the viral and host factors, especially the host immune system, may play a role in the chronicity of the HBV infection. Stimulator of interferon genes (STING) is one of the members of the pattern recognition receptor (PRR) that detects the presence of DNA in a human cell, activate synthesis of various cytokines and this protein is thought to be an important member of the immune system against HBV infection. Based on the assumption that there may be a relationship between the differences of STING expression in individuals and HBV chronicity, the aim of this study was to investigate STING gene expression levels in individuals naturally immunized against HBV, in chronic hepatitis B infected patients and in normal individuals who have not been exposed to HBV. A total of 90 volunteers have been included in the study from the age range of 18 to 65, in which the first group consists of 30 individuals naturally immunized against HBV, the second group consists of 30 chronically hepatitis B infected patients while the third group consists of 30 healthy population members who have not been exposed to HBV. Whole blood samples were taken from each participant and peripheral blood mononuclear cells (PBMC) were isolated afterwards. Total RNA was isolated from PBMC. After the synthesis of cDNA from the total RNA, STING gene expression levels were determined by real time polymerase chain reaction (Rt-PCR) method. Normalization was performed by applying the 2-ΔΔCT formula after Rt-PCR procedure. STING expression level of the naturally immunized group was calculated as 0.084 ± 0.026 on average, average STING expression level of healthy population group was 0.082 ± 0.032 and STING expression level of chronically infected patients group was 0.075 ± 0.022 on average. There was no statistically significant difference between the groups (p> 0.05). To our knowledge, this is the first study investigating the role of STING expression in the chronicity of HBV. Although there was no statistically significant difference between the groups, the data that showed STING expression levels in naturally immunized individuals were approximately 10% higher than those in chronic hepatitis B patients and was considered as an important finding.

The Prevalence of Mixed Genotype Infections in Turkish Patients with Hepatitis C: a Multicentered Assessment.

BACKGROUND: HCV virus infections are one of the major health problems in the world that can cause cirrhosis and liver cancer at a higher rate than other hepatitis data. The aim of this study was to determine the prevalence of mixed infections with different HCV genotypes in Turkey and also to evaluate the current HCV genotype and sub-type distributions by a multicentered assessment. METHODS: The HCV genotype data of 17,578 hepatitis C patients collected from 23 centers from different geographic regions covering all Turkey were collected. The data included information about the HCV genotypes in the last 10 years (between 2007 and 2016), demographic properties of the patients and the methods/systems used to determine the genotypes. RESULTS: Two hundred twenty-eight of the patients (1.3%) had mixed genotype. The most common mixed genotype combination was 1b + 4 (0.83%) followed by 1a + 1b (0.26%). Genotype distribution varies according to geographical regions. However, genotype 1 (82.92%) was the most common genotype in all regions and all years. This was followed by genotype 3 (7.07%) and genotype 4 (5.43%). A variety of methods were used by the centers including sequencing, pyrosequencing, real-time PCR, in-house RFLP, reverse hybridization (LIPA), and hybridization. CONCLUSIONS: Infection with mixed HCV genotypes in Turkey is uncommon. Genotype distribution varies according to geographic regions; the most common genotype 1 is encountered all over the country, while genotypes 3 and 4 are only in some of the centers. Since there is limited information about mixed HCV infection, further investigations are needed to determine the clinical importance of mixed HCV infection.

Determination of Seroprevalence of Borrelia burgdorferi IgG in Adult Population Living in Trabzon.

BACKGROUND: Lyme borreliosis is a tick-borne, multi-systemic infectious disease that is thought to be wide spread in Turkey even though studies on its seroprevalence are limited. AIMS: To determine the seroprevalence of Lyme borreliosis in part of north-eastern Tur-key (in the city of Trabzon), and to identify possible relationships between seropositivity and various factors such as location, gender, age group, occupation, income, and educational level. STUDY DESIGN: Retrospective cross-sectional study. METHODS: A total of 884 blood samples collected from provincial and district health centers serving a population of about 800,000 were included in this study. ELISA was used to determine the anti-Borrelia IgG antibody levels in the samples. Samples that yielded positive results by ELISA were further subjected to western blot (WB). RESULTS: IgG antibodies were found in 128 samples (14.5%). Statistical analysis revealed significant differences between age groups and educational levels in terms of the incidence of seropositivity, whereas location, gender, occupational group and income level had no effect (p<0.001, p<0.001, p=0.948, p=0.645, p=0.131, p=0.080 respectively). CONCLUSIONS: The risk of contracting Lyme borreliosis in Trabzon is high, and necessary measures need to be taken to avoid the spread of disease.

[Investigation of plasmid-mediated quinolone resistance genes in quinolone-resistant Escherichia coli and Klebsiella spp. isolates from bloodstream infections]. / Kan akimi enfeksiyonu etkeni olan kinolona dirençli Escherichia coli ve Klebsiella spp. izolatlarinda plazmid aracili kinolon direnç genlerinin arastirilmasi.

One of the treatment options of Escherichia coli and Klebsiella spp. infections which are the most common opportunistic pathogens of gram-negative sepsis is quinolones. Resistance to quinolones which act by disrupting DNA synthesis has been increasing. Horizontal transfer of plasmid-mediated quinolone resistance (PMQR) genes play an important role in the spread of resistance. The data about the prevalence of PMQR genes in our country is quite limited. The aim of this study was to investigate the presence of known PMQR genes namely qnrA, qnrB, qnrC, qnrS, qnrD, aac(6')-Ib-cr, qepA and oqxAB amongst quinolone-resistant E. coli and Klebsiella spp. strains isolated from blood cultures. One hundred twenty seven E.coli and 66 Klebsiella isolates detected as nalidixic acid- and/or ciprofloxacin-resistant by phenotypical methods, from 193 blood samples of 187 patients admitted to Karadeniz Technical University, Faculty of Medicine, Department of Medical Microbiology, Bacteriology Unit of Patient Service Laboratory between January 2012 to August 2013 were included in the study. The presence of PMQR genes were investigated by polymerase chain reaction (PCR) and for the detection of aac(6')-Ib-cr variants PCR-restriction fragment length polymorphism (PCR-RFLP) method was used. The positive bands were sequenced using the same primers, and aligned with formerly defined resistance gene sequences, and confirmed. In the study, 56.7% (72/127) of E.coli and 19.7% (13/66) of Klebsiella spp. isolates, with a total of 44% (85/193) of all the isolates were found to be phenotypically resistant to quinolones. Of the 13 resistant Klebsiella isolates, 11 were K.pneumoniae, and two were K.oxytoca. Extended-spectrum beta-lactamase (ESBL)-producing isolates showed higher resistance (50/80, 62.5%) to quinolones than the negative ones (35/113, 30.9%). The prevalence of quinolone resistance genes among resistant E. coli and Klebsiella spp. isolates was determined as qnrA, 1.4% and 15.4%; qnrB, 4.2% and 61.5%; qnrS, 1.4% and 7.7%; qepA, 1.4% (only E.coli); aac(6')-1b-cr, 38.9% and 92.3%; and oqxAB, 1.4% and 84.6%, respectively. qnrC and qnrD genes were not detected. Carriage of multiple resistance genes were observed more frequently among resistant Klebsiella isolates than E.coli strains. As a result, in this study investigating the contribution of transferrable genes to quinolone resistance, prevalence of PMQR genes in quinolone-resistant and blood isolates of E.coli and Klebsiella in our university hospital serving the region were found to be higher than the current data reported from the other studies in our country. Furthermore, this study presented the initial data for the first time in our country on the prevalence of qnrD which was undetected, and the frequency of oqxAB gene in clinical samples. However, location of oqxAB gene needs to be confirmed by conjugation or hybridization methods.

[Comparison of the modified Hodge test and the Carba NP test for detection of carbapenemases in Enterobacteriaceae isolates]. / Enterobacteriaceae izolatlarinda karbapenemazlarin saptanmasinda modifiye Hodge testi ve Carba NP testlerinin karsilastirilmasi.

A rapid, practical, and accurate identification of carbapenemase-producing Enterobacteriaceae isolates is crucial for the implementation of appropriate infection control measures and proper treatment of the infections. For this purpose, a large number of phenotypic test methods have been developed, although none has 100% sensitivity and specificity. Variations in sensitivity and specificity of these tests based on the type of beta-lactamase enzymes carried by that isolates might result in differences between regions and countries. The aim of this study was to compare the performances of widely used modified Hodge test (MHT) and Carbapenemase Nordmann-Poirel (Carba NP) test in the detection of carbapenemases in Enterobacteriaceae family members. A total of 65 Enterobacteriaceae isolates (43 bla(OXA-48), 10 bla(VIM), 9 bla(IMP), 1 bla(NDM-1), 1 bla(KPC-2) and 1 bla(OXA-48)+bla(VIM) carrying strains) that showed decreased sensitivity to at least one carbapenem (ertapenem, imipenem or meropenem), and carriage of carbapenemase gene confirmed by polymerase chain reaction (PCR), were included in the study. Seventy-eight isolates showing decreased susceptibility to carbapenems but lacking carbapenemase genes were used as controls. All isolates were identified by using conventional methods as well as automated BD Phoenix System (Becton Dickinson, USA). The antimicrobial susceptibility testing was performed using the same automated system, and was confirmed by disk diffusion method. Results were evaluated according to the CLSI criteria. MHT was performed in accordance with the CLSI guideline, and Carba NP test was carried out by a modified protocol. Instead of imipenem monohydrate, which was used in the original protocol, 6 mg/ml imipenem/cilastatin was used in the modified protocol. In the study, MHT identified 90.8% (59/65) of carbapenemase-producing isolates, while 93.9% (61/65) of the isolates were identified by Carba NP test. With MHT, four Klebsiella pneumoniae producing OXA-48, one Escherichia coli producing IMP, and one K.pneumoniae producing NDM-1, and with Carba NP test, one E.coli and one K.pneumoniae producing OXA-48, one E.coli producing IMP, and one Enterobacter cloacae producing VIM could not de detected. Three OXA-48-producing isolates (two K.pneumoniae and one E.coli) yielded late and weak positive results with Carba NP test. MHT had false positivity for 31 isolates, while Carba NP test showed no false positivity. In comparison of the sensitivity and specificity of the two tests, sensitivities were found to be similar although the Carba NP has a slightly higher sensitivity than the MHT (93.9% versus 90.8%, respectively; p= 0.754), Carba NP was found more specific (100% versus 60.3%, respectively; p< 0.0001). With Carba NP test, 26% of the isolates (n= 16) were positive within 15 minutes, and 85% (n= 52) were positive within the first hour. It was concluded that, Carba NP test showed high sensitivity and specificity than the MHT and the results can be obtained more rapidly for the presence of carbapenemases in Enterobacteriaceae. The use of MHT alone is not recommended to confirm the presence of carbapenemases produced by Enterobacteriaceae. On the other hand Carba NP test can be used for this purpose, however molecular analysis should be considered for suspicious negative results.

[Investigation of carbapenemases in carbapenem-resistant Escherichia coli and Klebsiella pneumoniae strains isolated in 2014 in Turkey]. / Türkiye'de 2014 yili içinde izole edilen karbapeneme dirençli Escherichia coli ve Klebsiella pneumoniae izolatlarinda karbapenemaz varliginin arastirilmasi.

Carbapenems are the choice of treatment in infections caused by multidrug resistant Enterobacteriaceae. In recent years carbapenem-resistant Enterobacteriaceae isolates due to carbapenemases have been increasingly reported worldwide. Multicenter studies on carbapenemases are scarce in Turkey. The aim of this study was to determine the distribution of carbapenemases from different parts of Turkey as a part of the European Survey of Carbapenemase Producing Enterobacteriaceae (EuSCAPE) project. Beginning in November 2013, carbapenem-resistant isolates resistant to at least one of the agents, namely imipenem, meropenem, and ertapenem were sent to the coordinating center. Minimum inhibitory concentrations for these carbapenems were determined by microdilution tests following EUCAST guidelines. Production of carbapenemase was confirmed by combination disk synergy tests. Types of carbapenemases were investigated using specific primers for VIM, IMP; NDM, KPC and OXA-48 genes by multiplex polymerase chain reaction. In a six month period, 155 suspected carbapenemase-positive isolates were sent to the coordinating center of which 21 (13.5%) were E.coli and 134 (86.5%) were K.pneumoniae. Nineteen (90.5%) strains among E.coli and 124 (92.5%) strains among K.pneumoniae were shown to harbour at least one carbapenemase gene by molecular tests, with a total of 92.3% (143/155). Carbapenemases were determined as a single enzyme in 136 strains (OXA-48: 84.6%; NDM: 6.3%; VIM: 2.8%; IMP: 1.4%) and as a combination in seven isolates (OXA-48 + NDM: 2.1%; OXA-48 + VIM: 2.1%; VIM + NDM: 0.7%). KPC was not detected in any of the isolates. According to the microdilution test results, resistance to imipenem, meropenem and ertapenem in OXA-48 isolates were 59.5%, 52.9% and 100%, respectively. The combination disk synergy test was 100% compatible with the molecular test results. As most of the OXA-48 producing isolates were susceptible to meropenem but all were resistant to ertapenem, ertapenem seems to be the most sensitive agent in screening carbapenemases in areas where OXA-48 is prevalent and phenotypic combination tests can be useful in centers where molecular tests are not available.

Serotype distribution of Streptococcus pneumoniae in children with invasive diseases in Turkey: 2008-2014.

Successful vaccination policies for protection from invasive pneumococcal diseases (IPD) dependent on determination of the exact serotype distribution in each country. We aimed to identify serotypes of pneumococcal strains causing IPD in children in Turkey and emphasize the change in the serotypes before and after vaccination with 7-valent pneumococcal conjugate vaccine (PCV-7) was included and PCV-13 was newly changed in Turkish National Immunization Program. Streptococcus pneumoniae strains were isolated at 22 different hospitals of Turkey, which provide healthcare services to approximately 65% of the Turkish population. Of the 335 diagnosed cases with S. pneumoniae over the whole period of 2008-2014, the most common vaccine serotypes were 19F (15.8%), 6B (5.9%), 14 (5.9%), and 3 (5.9%). During the first 5 y of age, which is the target population for vaccination, the potential serotype coverage ranged from 57.5 % to 36.8%, from 65.0% to 44.7%, and from 77.4% to 60.5% for PCV-7, PCV-10, and PCV-13 in 2008-2014, respectively. The ratio of non-vaccine serotypes was 27.2% in 2008-2010 whereas was 37.6% in 2011-2014 (p=0.045). S. penumoniae serotypes was less non-susceptible to penicillin as compared to our previous results (33.7 vs 16.5 %, p=0.001). The reduction of those serotype coverage in years may be attributed to increasing vaccinated children in Turkey and the increasing non-vaccine serotype may be explained by serotype replacement. Our ongoing IPD surveillance is a significant source of information for the decision-making processes on pneumococcal vaccination.

Effectiveness of transcranial magnetic stimulation application in treatment of tinnitus.

OBJECTIVE: Pathophysiology of tinnitus known to accompany nearly all disorders in hearing system has not been fully understood, and therefore, there are some difficulties in evaluation and treatment thereof. The objective of the current study is to research the effectiveness of transcranial magnetic stimulation (TMS) application in treatment of tinnitus. MATERIALS AND METHODS: Sixty patients aged between 15 and 70 years who applied to a polyclinic of Ear-Nose-Throat Department, Cerrahpasa Medical Faculty, Istanbul University, because of the complaint of tinnitus between January 2009 and July 2009 were selected using simple random sampling method. The treatment group and the placebo group were separated randomly as to include 30 patients. The tinnitus loudness, tinnitus frequency, tinnitus subjective score, and tinnitus handicap inventory results were compared before application of TMS and after 1 month. FINDINGS: It was found that the difference of tinnitus handicap inventory score within the TMS group (before the treatment: mean, 52.76; SD, 15.8; after the treatment: mean, 44.4; SD, 13.57) was statistically significant (P < 0.0001) and the difference of tinnitus handicap inventory score within the placebo group (before the treatment: mean, 51.46; SD, 15.41; after the treatment: mean, 51.13; SD, 16.86) was significantly insignificant (P = 0.848). When tinnitus severities determined at the beginning and end of the treatment were evaluated within each group, it was found that the difference of loudness within the group subjected to TMS was statistically significant (P < 0.0001) but the difference of loudness within the placebo group was statistically insignificant (P = 0.490). When tinnitus subjective scores were evaluated within each group before and after the treatment, the difference of subjective score within the group subjected to transcranial magnetic stimulation was statistically significant (P < 0.0001), and the difference of subjective score within the placebo group was statistically significant (P = 0.168). CONCLUSIONS: The studies showed that low-frequency repeating TMS is useful in the treatment of chronic tinnitus. In the current study performed on the patients with chronic tinnitus, it was shown that low-frequency repeating TMS has a statistically significant success when compared with the placebo group.

[Molecular epidemiology, antimicrobial resistance and characterization of extended-spectrum beta-lactamases of Salmonella enterica serotype Paratyphi B clinical isolates]. / Salmonella enterica serotip Paratyphi B klinik izolatlarinin moleküler epidemiyolojisi, antimikrobiyal direnci ve genislemis spektrumlu beta-laktamazlarinin karakterizasyonu.

Although Salmonella enterica serotype Paratyphi B is the less frequently isolated serotype worldwide and in Turkey, it is the most common serotype in our hospital, with a marked increase in 2007. The purpose of this study was to investigate the antibiotic susceptibility and the extended spectrum beta-lactamase (ESBL) profile, and molecular epidemiology of S. Paratyphi B isolates detected in our hospital microbiology laboratory. Seventy isolates identified as S. Paratyphi B from 109 Salmonella isolates obtained from clinical specimens from different patients between October 2005 and December 2012, were included in the study. In addition to conventional methods, isolates were identified using the Phoenix automated microbiology system (Becton Dickinson, USA). Serotyping of the isolates was performed on the basis of slide agglutination and the Kauffmann-White scheme. The antibiotic susceptibility of the isolates was determined using the BD Phoenix' automated system and disk diffusion test. ESBL enzymes were investigated using the combined disk test, isoelectric focusing, polymerase chain reaction (PCR) and sequence analysis. The molecular epidemiology of the 51 isolates obtained between October 2005 and August 2008 was examined with pulsed-field gel electrophoresis (PFGE) using the XbaI enzyme. S. Paratyphi B isolates were obtained from 70 specimens (46 blood, 16 fecal, 4 bone marrow, 2 urine and 2 wound) each from different patients. Resistance to nalidixic acid was determined in 18.6%, resistance to ampicillin, cefotaxime and cefepime in 2.9% and to ceftazidime and co-trimoxazole in 1.4% of the isolates. ESBL production was detected only in two isolates; in one TEM-1 was accompanied by CTX-M-15 and in the other isolate CTX-M-3 was found. Forty-six of the 51 isolates (90%) were found to be genetically related by PFGE and were placed in cluster A. The distribution of the isolates in cluster A revealed six subtypes as A1 (n= 7), A2 (n= 11), A3 (n= 7), A4 (n= 18), A5 (n= 2) and A6 (n= 1). Three different patterns not related to the cluster A were determined in the remaining five isolates (two were B, one of each was C, D and E). In conclusion, although the rate of antibiotic resistance was low in the S. Paratyphi B isolates in our hospital, rare types of ESBLs such as CTX-M-3 and CTX-M-15 were detected in Salmonellae. As far as the current literature is considered, this is the first report in Turkey of blaCTX-M-15 in Salmonella spp. and blaCTX-M-3 genes in S. Paratyphi B. The results may indicate a possible future threat to the treatment of Salmonella infections. Since most of the isolates were genetically related, this might suggest an epidemic in our region.

[Investigation of integrons, sul1-2 and dfr genes in trimethoprim-sulfametoxazole-resistant Stenotrophomonas maltophilia strains isolated from clinical samples]. / Klinik örneklerden izole edilen trimetoprim-sülfametoksazole dirençli Stenotrophomonas maltophilia suslarinda integron, sul1-2 ve dfr genlerinin arastirilmasi

Stenotrophomonas maltophilia, which is a non-fermentative gram-negative bacillus, has an increasing importance in nosocomial and opportunistic infections. Since it exhibits resistance to numerous broad-spectrum antibiotics such as aminoglycosides, beta-lactams and tetracyclines, it may considerably limit empirical treatment options. Trimethoprim-sulfamethoxazole (SXT) is recommended as the first-line therapy in the treatment of S.maltophilia infections thanks to its high potency and usefulness in a range of patients. In recent years, however, studies in different geographical regions have started to report resistance to SXT. In this study, we aimed to investigate the genes sul1, sul2, dfrA9, dfrA10, dfrA20 and class I, class II integron gene cassettes which are known to play role in SXT resistance among SXT-resistant S.maltophilia strains. A total of 618 S.maltophilia strains isolated from various clinical samples of 339 patients between January 2006 and October 2011 at the laboratory of Medical Microbiology Department, Faculty of Medicine, Karadeniz Technical University, Trabzon, Turkey, were included in the study. The isolates were identified by both conventional methods and the Phoenix automated identification system (Becton Dickinson, USA). SXT resistance was determined in the isolates of 32 patients (32/339, 9.4%) by both the automated system and agar dilution method of them 29 (90.6%) were hospital-acquired, and 3 (9.4%) were community-acquired. The genes which are known as SXT resistance determining genes including sul1, sul2, dfr genes, and class I and class II integron gene cassettes were analyzed by using specific primers with polymerase chain reaction in the 32 SXT-resistant isolates. Subsequently, nucleotide sequence analysis of the amplified materials was performed. As a result of this assay, the presence of class I integron gene cassette and sul1 gene were detected in one isolate. Nucleotide sequence analysis of the gene cassette revealed oxacilinase (oxa2) type of beta-lactamase, an aminoglycoside 6'-N-acetyltransferase [aac(6')-IIc], leading to resistance of aminoglycosides, and a quaternary ammonium compounds resistance gene (qacF), respectively. In conclusion, to best of our knowledge the sequences of class I integron gene cassette including oxa2, aac(6')-IIc, qacF genes were identified in S.maltophilia for the first time. It should be kept in mind that the co-presence of a class I integron gene cassette and the sul1 gene in S.maltophilia may lead to the development of multi-drug resistance and may act as a potential source for the dissemination of resistance.

[Epidemiology, clinical and microbiological characteristics of invasive streptococcal infections in Turkey, 2010-2011]. / Türkiye'de invazif streptokok enfeksiyonlarinin epidemiyolojisi, klinik ve mikrobiyolojik özellikleri, 2010-2011.

A one-year active surveillance study was conducted to investigate the epidemiological and microbiological characteristics of invasive group A streptococci (GAS) infections in Turkey and to provide data for the establishment of national preventive strategies related to invasive GAS infections. A total of 46 clinical microbiology laboratories from 12 different regions of Turkey (Istanbul; Eastern and Western Marmara; Eastern and Western Blacksea; Aegean; Mediterranean; Western, Central, Northeastern, Middle-eastern and Southeastern Anatolia) participated in the study. Accordingly, GAS strains isolated from sterile body sites (blood, cerebrospinal, synovial, pleural, peritoneal, pericardial fluids) in the study centers between June 2010-June 2011, were sent to Maltepe University Hospital Clinical Microbiology Laboratory for microbiological confirmation and further analysis. The isolates were identified by conventional methods, and for serotyping, opacity factor (OF) and T protein types were investigated. For genotyping GAS lysate preparation, emm gene amplification and sequencing were performed by using the protocols recommended by Centers for Disease Control and Prevention. A total of 65 invasive GAS strains were isolated in 15 of the participant centers, during the study period. The rate of invasive GAS isolation exhibited regional variation, with the highest rates in the Eastern Blacksea (Trabzon, n= 19), followed by Istanbul (n= 17) and Western Anatolia (Ankara, Konya, n= 14). Of the patients with invasive GAS infection 33 were female, 32 were male, with the age range of 0-89 years. GAS strains were most commonly isolated from soft tissue specimens (n= 18), followed by abscess material (n= 10), sterile body fluids (n= 8) and blood (n= 7) samples. Serotyping revealed that 55% (36/65) of the strains were OF positive, and the majority of T protein was polygroup T (n= 20), followed by U (n= 14), B (n= 5), X (n= 3) and Y (n= 2). T protein was not detected in 22 isolates. The strains were found to have 17 different emm types;emm1 (n= 13), emm4 (n= 6), emm6 (n= 6), emm12 (n= 6), emm24 (n= 4), emm14 (n= 3) and emm28 (n= 3). Nine of the strains could not be typed by sequencing. The correlation between emm typing and serotyping was detected as 58%. It was observed that 26-valent vaccines included 70.5% of the invasive GAS strains included in this study. Our study provided initial data concerning the epidemiological properties of invasive GAS infections and characterization of GAS strains in Turkey. The incidence of invasive GAS infections is low in our country. Although immunization programme by 26-valent GAS vaccine is not currently an urgent public health issue for our country, the results of this study indicated that emm types 4 and 24 should better be included in such a vaccine to be used in Turkey. Additionally, since epidemiological features of GAS infections and the microbiological characteristics of the strains can vary by time, for the diagnosis of invasive streptococcal infections and to take the necessary preventive measures, epidemiological studies should be conducted repeatedly.

[Determination of hepatitis C virus genotypes among hepatitis C patients in Eastern Black Sea Region, Turkey]. / Dogu Karadeniz Bölgesi Hepatit C Hastalarinda Hepatit C Virusu Genotiplerinin Belirlenmesi.

Hepatitis C virus (HCV), the major cause of transfusion-associated hepatitis, is an important public health problem in the world as well as in Turkey. HCV is grouped as six distinct genotypes and a large number of closely-related subtypes. Genotyping of HCV is an important tool for providing epidemiological data, prediction of prognosis, and optimization of antiviral therapy. This study was carried out to determine the distribution of HCV genotypes in hepatitis C patients residing in different provinces of the Eastern Black Sea Region, Turkey. A total of 304 HCV-RNA positive cases (151 male, 153 female; age range: 11-93 years, mean age: 55.2 ± 13.3 years) who were admitted to the Molecular Microbiology Unit of Department of Medical Microbiology, Karadeniz Technical University Faculty of Medicine, between January 2009 to December 2012, were included in the study. HCV genotypes were detected in plasma samples of the patients by using commercial assays [INNO-LiPA HCV II (Innogenetics, Belgium) or Abbott RealTime HCV Genotype II (Abbott Molecular Inc, USA)]. Due to the ambiguous genotyping results in some samples with these methods, an in-house multiplex polymerase chain reaction (PCR) assay with genotype-specific primers was also used in the study. Similar to the previous reports from Turkey, our results showed that four HCV genotypes (1, 2, 3, and 4) prevailed in the Eastern Black Sea Region and the predominant genotype and subtype were genotype 1 (92.8%) and 1b (87.5%), respectively. Distribution of genotypes were observed to vary according to the province. Prevalences of subtype 1a, genotype 2, 3, and 4 were noted as 5.3%, 1.6%, 4.9%, and 0.7%, respectively. Furthermore, the samples from Giresun, Gumushane and Bayburt provinces, which are relatively less immigrated, had higher genotype 1, and the prevalence rates in the region was affected by the presence of non-citizen residents. This study is the first report on distribution of HCV genotypes in chronic hepatitis C patients living in the provinces of Eastern Black Sea Region. Moreover, genotype-specific multiplex PCR assay could be useful in resolving certain methodological problems such as "ghost bands" encountered in line probe assay (LiPA) and multiple genotypes (including genotype 4) observed in real-time PCR during the characterization of HCV genotypes seen in Turkey.

Laryngeal foreign body mimicking croup.

Foreign body aspiration can be a fatal problem in all groups of ages. However, it is a leading cause of accidental deaths in children. Especially in the pediatric age group, diagnosis can be delayed because of various challenges. Children younger than 3 years carry the highest risk, as they are inclined to explore objects with their mouths. In most cases of inhaled foreign body, a positive history of aspiration is obtained. We are reporting a case of laryngeal foreign body that is mistreated for 1 week as a croup syndrome.