Gastrointestinal and Liver Manifestations in Children with COVID-19 and Their Relationship to Clinical Course.

OBJECTIVE: Coronavirus disease 2019 is a major health problem in all age groups. Although most clinical symptoms are respiratory, gastrointestinal symptoms are often reported. This is a major concern for children and has limited research coverage. In this study, we evaluated the frequencies of gastrointestinal symptoms and liver biochemical findings in children with coronavirus disease 2019 and their relationship with clinical course and length of hospital stay. MATERIALS AND METHODS: Demographic data, clinical, and laboratory findings of children with Coronavirus disease 2019 who were followed up by the Department of Pediatric Infectious Diseases between March 2020 and August 2020 were recorded. They were classified accord- ing to age groups as <5 years, 5-10 years, and >10 years. Laboratory findings were analyzed according to age groups. Demographic, clinical, and laboratory findings were compared in both situations, the presence of gastrointestinal symptoms and the presence of elevated liver enzymes. It was considered statistically significant if it was <.05. RESULTS: A total of 294 patients (median age 10 years [14 days to 18 years]) were enrolled in this study. Although fever is the most common symptom of coronavirus disease 2019, 15.6% of patients presented with acute gastroenteritis. Most patients with liver involvement (n = 130, 44.2%) were under 5 years of age (n = 74, 56.9%, P <.001). The patterns of abnormal liver test results were cholestatic (71.5%), hepatocellular (18.4%), and mixed (10%) types. Severe or mas- sive elevation of aminotransferase or liver failure was not observed. No statistically significant difference was noted in outcomes, including length of stay, for patients with gastrointestinal symptoms (P = .178) or liver involvement (P = .146). CONCLUSION: The presence of gastrointestinal symptoms or elevated liver enzymes does not affect the course of the disease in children with coronavirus disease 2019.

Reduced Monocyte Subsets, Their HLA-DR Expressions, and Relations to Acute Phase Reactants in Severe COVID-19 Cases.

Monocytes are one of the principal immune defense cells that encounter infectious agents. However, an essential role of monocytes has been shown in the spread of viruses throughout the human body. Considering this dilemma, this study aimed to evaluate monocyte subsets and Human Leukocyte Antigen-DR isotype (HLA-DR) expressions in clinical coronavirus disease 2019 (COVID-19) cases. This prospective, multicenter, case-control study was conducted with COVID-19 patients and healthy controls. The patient group was divided into two subgroups according to disease severity (severe and non-severe). Three monocyte subsets (classical, CL; intermediate, INT; non-classical, NC) were analyzed with flow cytometry upon the patients' hospital admission. A total of 42 patients with COVID-19 and 30 controls participated in this study. The patients' conditions were either severe (n = 23) or non-severe (n = 19). All patients' monocyte and HLA-DR expressions were decreased compared with the controls (p < 0.05). Per disease severity, all monocyte subsets were not significant with disease severity; however, the HLA-DR expressions of CL monocytes (p = 0.002) and INT monocytes (p = 0.025) were more decreased in the severe patient group. In patients with various clinical features, NC monocytes were more affected. Based on these results, NC monocytes were more decreased in acute COVID-19 cases, though related various clinics decreased all monocyte subsets in these patients. Decreased monocyte HLA expressions may be a sign of immune suppression in severe patients, even when the percentage of monocyte levels has not decreased yet.

Low level of antifungal resistance of Candida glabrata blood isolates in Turkey: Fluconazole minimum inhibitory concentration and FKS mutations can predict therapeutic failure.

BACKGROUND: Candida glabrata is the third leading cause of candidaemia in Turkey; however, the data regarding antifungal resistance mechanisms and genotypic diversity in association with their clinical implication are limited. OBJECTIVES: To assess genotypic diversity, antifungal susceptibility and mechanisms of drug resistance of C glabrata blood isolates and their association with patients' outcome in a retrospective multicentre study. PATIENTS/METHODS: Isolates from 107 patients were identified by ITS sequencing and analysed by multilocus microsatellite typing, antifungal susceptibility testing, and sequencing of PDR1 and FKS1/2 hotspots (HSs). RESULTS: Candida glabrata prevalence in Ege University Hospital was twofold higher in 2014-2019 than in 2005-2014. Six of the analysed isolates had fluconazole MICs ≥ 32 µg/mL; of them, five harboured unique PDR1 mutations. Although echinocandin resistance was not detected, three isolates had mutations in HS1-Fks1 (S629T, n = 1) and HS1-Fks2 (S663P, n = 2); one of the latter was also fluconazole-resistant. All patients infected with isolates carrying HS-FKS mutations and/or demonstrating fluconazole MIC ≥ 32 µg/mL (except one without clinical data) showed therapeutic failure (TF) with echinocandin and fluconazole; seven such isolates were collected in Ege (n = 4) and Gulhane (n = 3) hospitals and six detected recently. Among 34 identified genotypes, none were associated with mortality or enriched for fluconazole-resistant isolates. CONCLUSION: Antifungal susceptibility testing should be supplemented with HS-FKS sequencing to predict TF for echinocandins, whereas fluconazole MIC ≥ 32 µg/mL may predict TF. Recent emergence of C glabrata isolates associated with antifungal TF warrants future comprehensive prospective studies in Turkey.

[Investigation of Antibiotic Resistance and Virulence Factors of Enterococcus faecium and Enterococcus faecalis Strains Isolated from Clinical Samples]. / Klinik Örneklerden Izole Edilen Enterococcus faecium ve Enterococcus faecalis Izolatlarinin Antibiyotik Direnci ve Virülans Faktörlerinin Arastirilmasi.

Enterococci, which are commonly found in the environment, cause serious infections despite the absence of well-defined virulence factors and toxins. Knowing the virulence properties of enterococci is important to understand the complex pathogenic structures. In this study, we aimed to investigate the virulence factors (asa1, hyl, cylA, efa, ebp, ace, esp, gelE, sprE, fsrA, fsrB, fsrC genes, gelatinase activity, hemolysin, hydrogen peroxide and biofilm production) and antibiotic resistance of Enterococcus faecium and Enterococcus faecalis strains isolated from clinical specimens. A total of 110 enterococcus isolates which were accepted as infectious agents were included in the study. The polymerase chain reaction method was used to identify the isolates and to detect virulence genes. Characteristics of hemolysis, biofilm formation, hydrogen peroxide production and gelatinase activity were investigated by phenotypic methods. The antibiotic susceptibility test was performed with VITEK 2 automated system. E.faecalis ATCC 29212 standard strain was used as a quality control in all tests. Of the 110 enterococci isolates included in the study, 61 were identified as E.faecium and 49 as E.faecalis. The efa gene was the most frequently detected virulence gene (92.7%), followed by ace (83.6%), esp (66.4%), ebp (60.0%), cylA (50.9%), hyl (46.4%), asa1 (45.5%), gelE, sprE, fsrC (33.6%), fsrA (12.7%) and fsrB (11.8%). All genes except hyl were higher in E.faecalis isolates and the difference was statistically significant (p<0.05). Twenty-five (51%) E.faecalis and 1 (1.6%) E.faecium isolates had beta-hemolysis and the difference was statistically significant (p= 0.000). Seven (11.5%) E.faecium and 4 (8.2%) E.faecalis isolates formed biofilm, but the difference was not statistically significant (p> 0.05). Two (3.3%) E.faecium and 14 (28.6%) E.faecalis isolates exhibited gelatinase activity and the difference between the two species was statistically significant (p= 0.000). Hydrogen peroxide production was not detected in any of the isolates. The highest resistance rate was determined against ciprofloxacin (70.9%). The resistance to ampicillin was 69.1%, high level streptomycin 65.1%, high level gentamicin 39.4%, vancomycin and teicoplanin 4.5%, and linezolid 1.8%. In conclusion, our data indicated that virulence factors except hyl gene and biofilm production were higher in E.faecalis isolates but E.faecium isolates were more resistant to antibiotics. In order to prevent infection of such virulent or resistant isolates in the hospital setting, infection control measures must be followed. In vivo studies are needed for the better understanding of the virulence of enterococci.

Reference ranges for serum immunoglobulin (IgG, IgA, and IgM) and IgG subclass levels in healthy children

Background/aim: The serum immunoglobulin levels are used routinely in clinical practice because they provide key information on the humoral immune status. This study aimed to determine the age-related reference values of serum immunoglobulin levels in healthy children. Materials and methods: A total of 330 healthy children, aged between 0 and 18 years, were included in this study. The serum immunoglobulin levels were measured using a nephelometric method in a total of 11 groups, each group consisting of 30 individuals, and IgG subclasses in 6 groups of children aged more than 2 years. Results: The serum IgG levels were high during the newborn period, decreased until the sixth month, and again increased to a maximum level at the age of 18 years. The level of IgA was found to be extremely low in the newborn period and then increased with age. While the lowest value was in the newborn period for serum IgM level, the highest value was in the 16- to 18-year-old period. The IgG subclasses varied depending on the age groups. Conclusion: The updated reference intervals of immunoglobulin levels in children may be used for the accurate diagnosis of immune deficiencies.

[Investigation of Streptococcus pyogenes virulence factors and typing by multiple locus variable number tandem repeat fingerprinting (MLVF) method]. / Streptococcus pyogenes izolatlarinin virülans faktörlerinin arastirilmasi ve "multiple locus variable number tandem repeat fingerprinting (MLVF)" yöntemi ile tiplendirilmesi.

Streptococcus pyogenes is an important bacterial pathogen that colonizes the throat and skin of human beings and causes a wide variety of diseases ranging from mild infections like pharyngitis, tonsillitis and impetigo to severe invasive infections such streptococcal toxic shock syndrome, septicemia, and necrotizing fasciitis, and produces a wide variety of virulence factors. The aim of this study was to investigate the antibiotic resistance, virulence genes; [pyrogenic exotoxin genes (speA, C, G, H, I, J, K, L, M, smeZ and ssa), deoxyribonuclease genes (sdaB, spd3, sdc ve sdaD), protease genes (speB, spyCEP ve scpA) and inhibitor genes (mac and sic)] of S.pyogenes strains isolated from throat cultures of patients with symptomatic tonsillo-pharyngitis and typing by multiple locus variable number tandem repeat fingerprinting (MLVF) method. One hundred and fifty S.pyogenes isolates were identified by conventional methods and streptococcus group A latex kit (Biomerieux, France). Antibiotic susceptibility tests were performed by Kirby-Bauer disk diffusion method as recommended by Clinical and Laboratory Standards Institute. DNA isolation was performed by using a commercial DNA isolation kit (Qiagen, Germany) in accordance with manufacturer's recommendations. The virulence genes were determined by multiplex PCR. MLVF method was performed with multiplex PCR using specific primers for repeated sequences within bacterial genome. All of the S.pyogenes isolates were susceptible to penicillin G, cefotaxime, ceftriaxone, chloramphenicol, clindamycin, erythromycin, levofloxacin, vancomycin and linezolid. Among streptococcal pyrogenic exotoxin genes the most frequent gene was smeZ (90.0%) followed by speG (88.0%), speC (58.7%), ssa (42.7%), speA (33.3%), speJ (24.0%), speK (18.7%), speH (14.0%), speI (13.3%), speL and speM (9.3%). Of the DNase genes, sdaB was detected in all strains (100%), spd3, sdc, sdaD genes were determined as 64.7%, 36.0%, 24.7% respectively. Protease genes (speB, spyCEP, scpA) and mac gene from the inhibitor genes were positive in all strains, and sic gene was positive in only 3 (2.0%) of the isolates. Thirty-two different patterns that contained two or more isolates were determined by MLVF analysis. Ninety one isolates were included in any of the 32 different patterns, while 59 isolates were defined as sporadic isolates. In conclusion, S.pyogenes isolates collected from throat cultures of patients with symptomatic tonsillo-pharyngitis in Konya/Turkey were susceptible to all antibiotics studied and have carried a very high rate of virulence factors. However the isolates were mostly clonally unrelated and sporadic. This study is the first report in Turkey, in which S.pyogenes isolates were typed by the MLVF method and a large number of virulence factors were investigated.

[Investigation of various virulence factors of Klebsiella pneumoniae strains isolated from nosocomial infections]. / Hastane enfeksiyonu etkeni Klebsiella pneumoniae izolatlarinda çesitli virülans faktörlerinin arastirilmasi.

Klebsiella pneumoniae is an opportunistic pathogen that commonly affects immunosuppressed patients and causes nosocomial infections. K.pneumoniae has a variety of virulence factors, especially capsule polysaccharide, hypermucoviscosity (HV), fimbriae, toxins and determinants for iron acquisition. The aim of this study was to detect the virulence factors in K.pneumoniae strains isolated from nosocomial infections in two years. Fifty three K.pneumoniae strains isolated from the samples of patients with nosocomial infections in the Medical Microbiology Laboratory of Selcuk University Faculty of Medicine Hospital between 2011 and 2013 were included in the study. Identification and antimicrobial susceptibilities of the isolates were performed by VITEK 2 automatic system. Biofilm formation,α-hemolysin, capsule and HV were investigated by phenotypic methods. Polymerase chain reaction (PCR) was used to detect virulence genes encoding adhesins (fimH-1, mrkD, kpn, ycfM), siderophores (entB: enterobactin, iutA: aerobactin, irp-1, irp-2, ybtS, fyuA: yersiniabactin, iroN: catechols receptor), protectines or invasins (rmpA, magA, traT) and toxins (hlyA, cnf-1). Of the 53 K.pneumoniae isolates,12 (22.6%) were isolated from in patients of reanimation intensive care unit, 8 (15.1%) medical oncology, 7 (13.2%) newborn intensive care unit and 26 (49%) other clinics. The distribution of the isolates according to the samples was as follows: urine (n= 14), blood (n= 13), wound (n= 8), drainage fluid (n= 10), broncho-alveolar lavage (n= 7), and cerebrospinal fluid (n= 1). Isolates which were resistant to meropenem were 5.7% and production of extended spectrum beta-lactamase (ESBL) was 71.7%. The capsule, biofilm formation, and HV were observed in 100%, 79.2%, and 1.9% of the isolates, respectively. Production of α-hemolysin was not detected in any of the isolates. The genes; entB (96.2%), ycfM (86.8%), and mrkD (83.0%) showed high prevalence. The other genes were detected in different ratios: fimH-1 (64.2%), fyuA (54.7%), kpn (49.1%), ybtS (41.5%), irp-1(41.5%), irp-2 (37.7%), traT (11.3%) and iutA (5.7%). Virulence genes; iroN, rmpA, magA, hlyA and cnf-1 were not detected in any of the isolates. Enterobactin had the highest rate among siderophores, and ycfM and mrkD in adhesins. The capsule and biofilm formation were commonly found in the isolates. Hypermucoviscosity was only found in one isolate but associated genes were not detected. Alfa hemolysin production and hlyA gene were not determined. As a result, it seems that the basis of the pathogenicity of K.pneumoniae strains isolated from nosocomial infections are capsule, adhesins, enterobactin and ability of biofilm formation. There is a need for new studies for the continuous monitoring of toxin and invasion ability as well as antibiotic resistance in the control of hospital infection caused by K.pneumoniae.

Tigecycline Therapy in an Infant for Ventriculoperitoneal Shunt Meningitis.

Shunt infections are seen in 3% to 20% of patients who have cerebrospinal fluid (CSF) shunts. Although the staphylococcal species are the most common cause of shunt-related infections, Gram-negative bacteria are increasingly reported with higher mortality rates. Tigecycline, a glycylcycline, is not approved for children. But in the era of nosocomial infections due to multidrug-resistant pathogens, it can be the life-saving option. We report an infant with ventriculoperitoneal shunt-related meningitis treated with a tigecycline combination regimen. A 5-month-old boy who had a ventriculoperitoneal shunt was admitted with meningitis. Extended spectrum ß-lactamase-producing Klebsiella pneumoniae grew in the CSF. At the end of the fourth week of intravenous meropenem plus gentamicin therapy, carbapenem-resistant K pneumoniae grew in the CSF (mean inhibitory concentration value for meropenem >4 µg/mL, by E-test). The infected shunt was removed, and an external ventricular drainage catheter was inserted. With permission, intravenous tigecycline (1.2 mg/kg per dose twice a day) and intrathecal amikacin were added to the meropenem. Intrathecal amikacin could be given for only 7 days. On the sixth day of tigecycline treatment, the CSF was sterilized. Antibiotic therapy was given and consisted of a total of 60 days of meropenem and 20 days of tigecycline therapy. Because no available efficacy and safety data from randomized-controlled studies exist, tigecycline must be used only as salvage therapy, in combination with other drugs, for critically ill children who have no alternative treatment options.

[Investigation of OXA type beta-lactamases and PFGE patterns in Acinetobacter baumannii strains resistant to carbapenems]. / Karbapenemlere dirençli Acinetobacter baumannii suslarinda OXA tipi beta-laktamazlarin arastirilmasi ve PFGE ile genotiplendirilmesi.

Acinetobacter baumannii is an important opportunistic and multidrug-resistant pathogen leading to nosocomial infections. Over the last 10 years, a significant and threatening increase in resistance to carbapenems, mainly due to the dissemination of class D beta-lactamases, has been reported in A.baumannii worldwide. The most common types of beta-lactamases causing carbapenem resistance in A.baumannii are the OXA-23, OXA-24, OXA-40, OXA-58 and OXA-143 type serine beta-lactamases. The aim of this study was to investigate the presence of OXA type beta-lactamases in carbapenem-resistant A.baumannii strains and the clonal relationship between the strains. A total of 105 non-duplicate carbapenem-resistant A.baumannii strains isolated from various clinical samples (68 blood, 18 bronchoalveolar lavage, 13 drainage, 3 urine, 2 cerebrospinal fluid and 1 catheter samples) in the Microbiology Laboratories of Selcuk University, Meram (2009-2012) and Selcuklu (2007-2008) Medical School Hospitals, were included in the study. The isolates were identified by conventional methods and Phoenix 100 BD (BD Diagnostic, USA) and Vitek II (bioMerieux, France) automated systems. Carbapenem susceptibility test was performed by Kirby-Bauer disk diffusion method according to the CLSI standards. bla(OXA 23-like), bla(OXA 24-like), bla(OXA 58-like) and bla(OXA 51-like) genes were amplified by multiplex PCR assay and clonal relatedness was investigated by pulsed-field gel electrophoresis (PFGE) using ApaI enzyme. The bla(OXA 51-like) gene was determined in all carbapenem-resistant A.baumannii isolates, while the bla(OXA 23-like) and bla(OXA 58-like) genes were detected in 46.6% and 53.3% of isolates, respectively. However bla(OXA 24-like) gene was not demonstrated in any isolates. bla(OXA 23-like) gene was determined in both Meram and Selcuklu Medical School hospitals, but bla(OXA 58-like) gene was detected only in Meram Medical School hospital. PFGE analysis of the isolates revealed 32 different groups in bla(OXA 23-like) producing A.baumannii strains and 23 different groups determined in bla(OXA 58-like) producing strains. No common epidemic isolates were detected in the two hospitals, however it was noted that some clones produced small outbreaks in Meram MS hospital. In this study it was shown that bla(OXA 23-like) and bla(OXA 58-like) genes together with bla(OXA 51-like) gene had significant roles in the carbapenem-resistance of A.baumannii strains. Carbapenem-resistant A.baumannii strains producing bla(OXA 23-like) and bla(OXA 58-like) enzymes showed the epidemic potential of this nosocomial pathogen and the requirement of molecular typing methods to identify the epidemiologic relationship of the isolates.

[In vitro synergistic activity of sulbactam in combination with imipenem, meropenem and cefoperazone against carbapenem-resistant Acinetobacter baumannii isolates]. / Karbapeneme dirençli Acinetobacter baumannii izolatlarina karsi sulbaktam ile imipenem, meropenem ve sefoperazon kombinasyonlarinin in vitro sinerjistik aktivitesi.

Acinetobacter baumannii which is an opportunistic pathogen leading to nosocomial epidemics, exhibit high rates of antimicrobial resistance. Treatment of Acinetobacter infections is a challenge since most of the isolates are multiple antibiotic resistant. The aim of this study was to investigate minimum inhibitory concentrations (MICs) of sulbactam, imipenem, meropenem, and cefoperazone and in vitro synergistic activity of sulbactam in combination with imipenem, meropenem and cefoperazone against A.baumannii isolates of hospitalized patients. Forty A.baumannii strains isolated from various clinical specimens and found to be resistant to carbapenems by disc diffusion method, were included in the study. The isolates were identified by conventional methods and VITEK 2 (bioMerieux, France) automated identification system. MICs of sulbactam, imipenem, meropenem, and cefoperazone were determined by the broth microdilution method according to the standards of CLSI and in vitro synergy test was performed using the checkerboard microdilution method. Synergistic, partial synergistic, additive, indifferent and antagonistic effects of drug combinations were evaluated with the fractional inhibitory concentration index (FICI). Interpretation of the FICI was as follows: ≤ 0.5 synergy; > 0.5 to < 1 partial synergy; 1 additive; > 1 to < 4 indifference; and ≥ 4 antagonism. Forty A.baumannii isolates were resistant to imipenem and cefoperazone, but two were susceptible, seven were moderately susceptible and 31 were resistant to meropenem with the microdilution method. MIC values of the isolates for sulbactam were found to be 4 µg/ml in two, 8 µg/ml in five, 16 µg/ml in three, 32 µg/ml in 13, 64 µg/ml in three, 128 µg/ml in six and > 128 µg/ml in eight isolates. According to the FICI; imipenem/sulbactam combination exhibited synergy in 18 (45%), partial synergy in 4 (10%) and indifferent effect in 2 (5%) isolates, the combination of meropenem and sulbactam showed synergy in 19 (48%), partial synergy in 3 (7.5%), and indifferent effect in 3 (7.5%) isolates, the combination of cefoperazone/sulbactam demonstrated synergy in 18 (45%), partial synergy in 2 (5%), and indifferent effect in 2 (5%) isolates. There was no antagonistic effect with the tested combinations. In conclusion, MIC values of sulbactam were generally high in carbapenem-resistant A.baumannii strains. However, synergistic effect was detected in approximately half of the strains with the sulbactam/carbapenem combinations. The data obtained in this study should be supported by further advanced in vitro and clinical studies to predict the accurate clinical efficacy of sulbactam containing combinations on A.baumannii infections.

Two outbreaks of ESBL-producing Klebsiella pneumoniae in a neonatal intensive care unit.

BACKGROUND: In the present study, two epidemic episodes of extended spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae in the neonatal intensive care unit (NICU) were evaluated. METHODS: Routine and surveillance culture samples were taken from seven neonates with signs of infection in the NICU of Selcuk University Faculty of Medicine between 10 March and 25 April 2011, and between 11 June and 30 September 2011. RESULTS: ESBL-producing K. pneumoniae strains were isolated in six different samples (one wound, one blood, and four cerebrospinal fluid cultures) of the three neonates in the first episode and in 11 different samples (seven blood and four cerebrospinal fluid cultures) of the four neonates in the second episode. ESBL-producing K. pneumoniae was isolated from inguinal, axillar region, and stool samples of the nine colonized neonates in the second episode. It was determined on pulse field gel electrophoresis that all strains originated from two clones. CONCLUSIONS: The deficiencies in the infection control measures in an NICU may transform into an epidemic rapidly. Therefore, periodic training, observation, and monitoring of compliance are important.

[MLST types of vancomycin-resistant Enterococcus faecium strains isolated from blood cultures]. / Kan kültürlerinden izole edilen vankomisine dirençli Enterococcus faecium suslarinin MLST tipleri.

Enterococci, particularly vancomycin-resistant enterococci (VRE), are important nosocomial pathogens with limited treatment options. Enterococci have low-level resistance to penicillins and aminoglycosides and are intrinsically resistant to cephalosporins. In addition, they can acquire high-level resistance to beta-lactam antibiotics, aminoglycosides and glycopeptides. The aim of this study was to determine glycopeptide resistance mechanisms and genetic relationships of vancomycin-resistant E.faecium strains isolated from blood cultures between 2003-2009 years by molecular epidemiologic methods. A total of 38 VRE strains isolated from blood cultures were included in this study. The isolates were identified by conventional methods and Phoenix 100 BD automated system (Becton Dickinson Diagnostic Systems, USA) and confirmed by sequence analysis of 16S rRNA amplicons. Antibiotic susceptibility tests were performed by the Kirby-Bauer disk diffusion method accor-ding to the CLSI standards. MIC values of vancomycin were determined in vancomycin resistant strains by E-test (AB Biodisk, Sweden) method. Vancomycin resistance genes included vanA, vanB, vanC, and vanD were investigated by polymerase chain reaction (PCR) method. Clonal relationship between strains was determined by pulsed-field gel electrophoresis (PFGE). Sequence analysis was performed for examples selected for multilocus sequence typing (MLST) of each pulsotype and subtype. Thirty eight strains of enterococci isolated from blood cultures were defined as E.faecium by phenotypic methods and confirmed by 16S rRNA sequence analysis. Vancomycin MIC values of strains were determined as > 256 µg/ml by E test. The vanA gene was detected in all isolates. Clonal relationship of 38 isolates E.faecium carrying the vanA gene was determined by PFGE and MLST methods. PFGE detected four pulsotypes (A-D) and one sporadic isolate. Twenty nine strains belonged to A pulsotype, three strains belonged to B pulsotype, two strains belonged to C pulsotype and three strains belonged to D pulsotype. Out of 29 isolates, eight strains were type A1, nine strains were type A2, six strains were type A3, two strains were type A4 and four strains were type A5. MLST identified four different sequence types (STs). Twenty nine A pulsotype and its subtypes belonged to ST117 (76.3%), three B pulsotype belonged to ST280 (7.9%), two C pulsotype belonged to ST18 (5.2%) and three D pulsotype belonged to ST17 (7.9%). In conclusion, bloodstream infections caused by VRE in our hospital arose from a dominant strain belonged to ST117. However, presence of different pulsotypes of this strain indicated that the strain had been present in the hospital for a long time and had accumulated genetic variations. In addition, infections caused by minor pulsotypes were also detected. Therefore for prevention and control of the spread of nosocomial infections caused by VRE, it is crucial to identify resistance patterns and clonal relationship of these organisms.

[Distribution of emm genotypes and antibiotic susceptibility of Streptococcus pyogenes strains: analogy with the vaccine in development]. / Streptococcus pyogenes suslarinin emm genotiplerinin dagilimi ve antibiyotik duyarliligi: Gelistirilmekte olan asi ile karsilastirma.

Streptococcus pyogenes is the most common bacterial pathogen causing pharyngotonsillitis, and also can lead to diseases such as otitis media, impetigo, necrotizing fasciitis, bacteremia, sepsis and toxic shock-like syndrome. M protein encoded by emm gene is an important virulence factor of S.pyogenes and it is used for genotyping in epidemiological studies. The aims of this study were to determine the M protein types of group A streptococci (GAS) by using emm gene sequence analysis method, to compare the M types in terms of analogy with the vaccine in development and to determine the antibiotic susceptibilities of the isolates. A total of 35 GAS strains isolated from various clinical specimens in our laboratory were included in the study. Strains growing in blood culture were considered as invasive, strains growing in throat and abscess cultures were considered as non-invasive. The isolates have been identified by conventional methods and 16S rRNA sequence analysis at species level. emm genotyping of strains identified as S.pyogenes, was performed by PCR method as proposed by the CDC. Amplicons were obtained and sequenced in 23 out of 35 isolates. The results were compared with CDC emm sequence database. Antibiotic susceptibility of the isolates was performed by agar dilution method and evaluated as recommended by CLSI. Twenty-three out of 35 isolates could be typed and 15 different emm genotypes were detected. The most common emm types were emm1 (22%), emm89 (13%), emm18 (9%) and emm19 (9%). The detection rate of other emm types (emm5, 12, 14, 17, 26, 29, 37, 74, 78, 92, 99) was 47%. Types emm1, 12, 19, 74, 89 and 99 were observed in strains isolated from blood cultures. It was detected that nine of the 15 (60%) emm types are within the contents of 26 valent vaccine (emm 1, 5, 12, 14, 18, 19, 29, 89, 92). It was also observed that 17 (74%) of the 23 cases were infected by vaccine types and the four emm types (emm1, 12, 19, 89) identified in blood samples were among the vaccine types. All of the strains were found susceptible to penicillin, ampicillin, erythromycin, lincomycin, gentamicin, chloramphenicol, vancomycin and linezolid, however six isolates were resistant to levofloxacin (MIC= 4 and 16 µg/ml) and one isolate was resistant to tetracycline (MIC= 16 µg/ml). In conclusion, this preliminary local study with limited number of invasive and non-invasive S.pyogenes isolates, emphasized the need for larger scale multi-center studies to determine the analogy and efficacy of the vaccine in development.

[Determination of hemolytic activity and in vitro antifungal susceptibility of Trichophyton rubrum clinical strains]. / Trichophyton rubrum Klinik Izolatlarinin Hemolitik Aktivitesi ve Antifungal Ilaçlara In Vitro Duyarliliginin Saptanmasi

Trichophyton spp. which are among the agents of dermatophytosis with high morbidity, produce many virulence factors including hemolysins that exhibit toxic activity on immune system cells. Since relapses and chronicity are common problems related to dermatophytosis, prompt and appropriate treatment is of crucial importance. However, treatment is getting difficult due to the choice of inappropriate antifungals and increasing rates of cross-resistance among antifungal agents. The aims of this study were to investigate the hemolytic activities of Trichophyton rubrum strains isolated from patients with dermatophytosis and to detect the in vitro susceptibilities of those strains to ketoconazole, itraconazole, sulconazole, econazole and terbinaphine. Hair, skin and nail samples of patients were examined with direct microscopy using potassium hydroxide and cultivated on mycobiotic agar and Sabouraud dextrose agar. To determine hemolytic activities of T.rubrum strains, they were subcultured in Columbia Agar with 5% sheep blood and incubated for 7-14 days at 25°C in aerobic conditions. Media which displayed hemolysis were further incubated for 1-5 days at 37°C to increase hemolytic activity. Antifungal susceptibility testing was done with broth microdilution method guided by Clinical and Laboratory Standards Institute (CLSI) M38-A document. A total of 79 T.rubrum strains which exhibited negative urease and hair perforation tests, yielded pigmentation in potato-dextrose agar, were evaluated in the study. Hemolytic activity was detected in 71 strains (89.9%). Fifty strains showed incomplete (alpha) hemolysis and 21 strains showed complete (beta) hemolysis, whereas hemolysis was absent in eight of the isolates. Larger colonies created a larger zone of hemolysis and the smaller ones created a smaller zone. However, alpha-hemolysis did not turn to beta-hemolysis following further enlargement of the colony. According to antifungal susceptibility testing, the minimum inhibitory concentration (MIC) ranges, MIC50 and MIC90 values were found 0.0125-4 µg/ml, 0.5 and 2 µg/ml for ketoconazole; 0.0625-2 µg/ml, 0.5 and 1 µg/ml for itraconazole; 0.0313-4 µg/ml, 0.25 and 1 µg/ml for sulconazole; 0.0313-0.125 µg/ml, 0.0313 and 0.0625 µg/ml for econazole; 0.0313-0.0313 µg/ml, 0.0313 and 0.0313 µg/ml for terbinaphine, respectively. When the MIC values of hemolytic and non-hemolytic T.rubrum strains were compared, it was detected that hemolytic activity had no effect on MIC values. Our data have indicated that terbinaphine was the most effective antifungal agent against T.rubrum, while MIC values for itraconazole which is in common clinical use, were higher than expected and MIC values for econazole were lower than expected.

[Investigation of parvovirus B19 seroprevalence in various age groups in Central Anatolia Region, Turkey]. / Konya bölgesinde çesitli yas gruplarinda parvovirus B19 seroprevalansinin arastirilmasi

Human parvovirus B19 is a small, non-enveloped, icosahedral symmetric, single-stranded DNA virus that can cause a number of diseases, notably erythema infectiosum in children and aplastic crisis in patients with chronic hemolytic disorders. There have been limited data on the epidemiological pattern of parvovirus B19 infection in Turkey. The objective of this study was to investigate the seroprevalence of parvovirus B19 in Konya province (Central Anatolia), Turkey. Parvovirus B19 IgG antibodies were investigated by a commercial ELISA kit (RIDASCREEN, R-Biopharm AG, Germany) in 631 adults (age range: 18-> 60 years) and 542 children (age range: 0-17 years). The overall prevalence of parvovirus B19 IgG antibodies was 28.9%. The rate of parvovirus B19 IgG positivity was 20.7% (112/542) in the 0-17 years age group and was 36% (227/631) in the adult population. No significant difference in seropositivity rates were detected in terms of sex in children and adult group (p>0.05 in both groups). The rates of parvovirus B19 IgG seropositivity were 15.8% in 0-4 years age group, 16% in 5-9 years, 24.2% in 10-14 years, 40.9% in 15-19 years, 34.7% in 20-29 years, 35.5% in 30-39 years, 32.2% in 40-49 years, 37.5% in 50-59 years and 53.8% in > 60 years age group. The seropositivity rates in 0-4 and 5-9 years age groups were lower than the other age groups and the difference was statistically significant (p< 0.05). To determine the prevalence of parvovirus B19 in different age groups in different geographical areas is necessary since this will provide important information about the epidemiology of such infections.