[Heterogeneous macrolide resistance in methicillin-resistant Staphylococcus aureus isolates: investigation of resistance mechanisms and clonality]. / Metisiline dirençli Staphylococcus aureus izolatlarinda heterojen makrolid direnci: Direnç mekanizmasinin ve klonalitenin arastirilmasi

Staphylococcus aureus is one of the most important pathogens leading to hospital infections and has ability to gain multiple resistance to most of the antibiotics used as well as ability to spread clonally. In this study we aimed to investigate the mechanism of heterogenous macrolide resistance and clonality of 41 methicillin-resistant S.aureus (MRSA) strains isolated from clinical samples of inpatients between 2006 and 2008 in a tertiary care training hospital. Heterogenous macrolide resistance is defined as the isolates with colonies in the inhibiton zone of erythromycin, azithromycin, claritromycin discs, and named as hMLSB (heterogeneous macrolides, lincosamides, streptogramin-B) phenotype. A total of 63 macrolide-resistant MRSA strains isolated during the same period with non-hMLSB phenotype were included in the study as a control group. The susceptibilties of isolates to methicillin, erythromycin, azithromycin, claritromycin, clindamycin, quinupristin-dalfopristin, penicillin, vancomycin, teicoplanin, linezolide, gentamicin, amikacin, ciprofloxacin, trimethoprim-sulphamethoxazole, chloramphenicol, rifampin, tetracycline and telithromycin were determined by disc diffusion method according to the CLSI criteria. hMLSB isolates were also tested for erythromycin, clindamycin and quinupristin-dalfopristin susceptibility by Vitek-2 automated system (bioMerieux, France). Presence of erm(A), erm(B), erm(C), msr(A/B) resistance genes were investigated by PCR. The regulatory region of the erm(A) gene, which was found to be the only molecular mechanism of hMLSB, was sequenced. Sequence analysis of regulatory regions showed deletion of "guanine" base at position 149 in 34 strains and two point mutations namely "C368G" and "T377C" in 16 strains. Pulsed-field gel electrophoresis (PFGE) analysis indicated presence of a common clone and its variants; however this clone was also common among control group strains. Interestingly all heterogeneous macrolide resistant isolates were reported as susceptible to erythromycin, clindamycin, quinupristin-dalfopristin by the automated system. As a result a clone specific to heterogenous macrolide resistant strains was not detected. Although all hMLSB strains had erm(A) gene, a specific and common genetic modification could not be found in regulatory region of the isolates. The most important finding of this study is the detection of inability of the automated system to properly reflect the hMLSB phenotype. In addition it was suggested that longer incubation (at least 24 hours) of the antibiotic susceptibility test plates could help identification of hMLSB phenotype in disc diffusion tests.

[Investigation of extensive drug resistance in multidrug resistance tuberculosis isolates]. / Çok Ilaca Dirençli Tüberküloz Izolatlarinda Yaygin Ilaç Direncinin Arastirilmasi

Increasing number of drug resistant tuberculosis (TB) cases, observed in recent years, is an important public health problem. Extensively drug resistant TB (XDR-TB) is the development of resistance against any fluoroquinolones and at least one of the injectable second line anti-TB drugs in addition to resistance against isoniazide and rifampicin which are the first line anti-TB drugs [definition of multidrug resistant TB (MDR-TB)]. Anti-TB therapy failed with first-line anti-TB drugs due to MDR-TB cases is being planned according to second-line anti-TB drug susceptibility test results if available and if not, standart treatment protocols are used. Although it is recommended that individual anti-TB therapy should be designed according to the isolate's susceptibility test results, standart therapeutic protocols are always needed since second-line anti-TB drug susceptibility testing generally could not be performed in developing countries like Turkey. For this reason, nationwide and regional surveillance studies to determine the resistance patterns are always needed to make decisions about the standard therapy algorithms. In this study, it was aimed to investigate the presence of extensive drug resistance among 81 MDR-TB isolates obtained from various health care facilities from Istanbul, Izmir and Manisa and to determine the XDR-TB incidence in Marmara and Aegean regions. Furthermore, we aimed to provide epidemiological data to clinicians to support their choice of second-line anti-TB drugs for MDR-TB infections. Susceptibility testing of isolates for the first and the second-line anti-TB drugs were performed by using modified Middlebrook 7H9 broth in fluorometric BACTEC MGIT 960 system (Becton Dickinson, USA). Eighty-one MDR-TB isolates included in this study were isolated from 43 (53.1%) patients residing in Istanbul, 26 (32.1%) in Izmir and 12 (14.8%) in Manisa provinces. We could not find any isolate consistent with XDR-TB definition in this study. Second-line drug resistance rates of MDR-TB isolates to amikacin and kanamycin were 1.2%, ofloxacin and levofloxacin were 2.5%, capreomycin was 14.8%, ethionamide was 37% whereas linezolid resistance was not detected. Statistically significant correlation was detected between resistance rates of these antibiotic pairs; levofloxacin-ofloxacin (p< 0.01), amikacin-kanamycin (p= 0.01) and streptomycin-ethionamide (p= 0.04). In our study, extensive drug resistance was not encountered in any MDR-TB isolates while high resistance rates was observed against ethionamide and capreomycin. It can be concluded that parenteral aminoglycosides amikasin and kanamycin, fluoroquinolones and linezolid seemed to be reliable anti-TB agents in MDR-TB treatment, however, further larger scale studies are needed.

[Investigation of a nosocomial outbreak caused by ESBL positive Klebsiella pneumoniae in neonatal intensive care unit by AP-PCR]. / Yenidogan Yogun Bakim Ünitesinde Ortaya Çikan GSBL Pozitif Klebsiella pneumoniae Nedenli Nozokomiyal Bir Salginin AP-PCR ile Arastirilmasi

In the present study, we describe an outbreak caused by extended-spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae in the neonatal intensive care unit (NICU) of a tertiary-care hospital. Nosocomial blood-stream infections were detected in three patients hospitalized in NICU. Two of the cases were transferred to NICU due to premature birth and the other due to the presence of cleft palate and retrognathia. K.pneumoniae was isolated on the 5th day of hospitalization of the first patient from umbilical swab and blood culture; on the 15th day of hospitalization of the second patient from blood culture and on the 7th day of hospitalization of the third patient from blood culture. The isolates were identified by automated API Rapid ID 32 Staph (BioMerieux, France) system in addition to conventional laboratory methods. Antibiotic susceptibilities of the isolates were determined by using Kirby-Bauer disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) criteria. The same antibiotic susceptibility patterns were detected in all isolates. Active surveillance cultures included environmental sampling from surfaces of NICU, laryngoscopes, ventilators and connections of ventilators, stethoscopes, nebulizers, aspiration tubings, disinfectant solutions, couveuse and couveuse distilled water. Two ESBL producing K.pneumoniae strains, presenting the same antibiotic susceptibility pattern with the clinical strains, were isolated from one couveuse distilled water sample and one aspiration tubing. All of the K.pneumoniae isolates were resistant to amoxycillin-clavulonic acid, cefazolin, cefepime, ceftriaxone, ceftazidime, cefuroxime, aztreonam and trimetoprim-sulphametoxazole and susceptible to cefoxitin, imipenem, meropenem, gentamicin, tobramycin, amikacin, netilmisin, tetracycline, ciprofloxacin and chloramphenicol. Arbitrarily primed polymerase chain reaction (AP-PCR) analysis done with M13 primer revealed the same genotype for the patient and environmental K.pneumoniae isolates. It was concluded that AP-PCR which is a simple, rapid and cheap method for the determination of genetic relatedness between isolates, can be applied for the detailed evaluation of nosocomial outbreaks to detect the source of infection and control the dissemination of the outbreak.

Co-expression of plasmid-mediated quinolone resistance-qnrA1 and blaVEB-1 gene in a Providencia stuartii strain.

An extended-spectrum B-lactamase (ESBL)-producing Providencia stuartii isolate was studied. A qnrA1 gene co-expressing blaVEB-1 gene was detected. Both genes were transferred to the recipient strain. The ciprofloxacin MIC of recipient strain increased tenfold. The blaVEB-1 gene persisted in microorganisms in Turkey but it also spread with PMQR genes to other species. The combination of PMQR with multidrug resistant isolates producing ESBLs may compromise the use of valuable antibiotics. Serious efforts are necessary to detect PMQR determinants not only with common B-lactamases in widespread pathogens but also with uncommon forms that are encountered infrequently.

[Phenotypic and genotypic identification of Candida strains isolated as nosocomial pathogens]. / Hastane enfeksiyonu etkeni olarak izole edilen Candida suslarinin genotipik ve fenotipik olarak tanimlanmasi.

Over the last decade, there have been important changes in the epidemiology of Candida infections and antifungal agents used to treat these infections. In recent years, Candida species have emerged as important causes of invasive infections among patients in intensive care units. One of the main goals of this study was to evaluate the molecular epidemiology of infectious Candida species isolated in our hospital and accordingly supply data for hospital infection (HI) control. The other aim of this study was to evaluate effectiveness and practical applicability of traditional and molecular methods used to identify Candida isolates to the species level. A total of 77 Candida strains that were isolated from various clinical specimens of 60 hospitalized patients (29 male, 24 female; 7 were children) were included in the study. Fifty-seven (74%) of those isolates were defined as HI agents according to Centers for Disease Control and Prevention (CDC) criteria. The most common Candida species identified as agents of HI were C.albicans (22; 38.6%), followed by C.tropicalis (14; 24.6%), C.parapsilosis (13; 22.8%), C.glabrata (7; 12.3%) and Candida spp. (1; 1.75%). It was determined that bloodstream (26; 45.6%) and urinary tract infections (24; 42.1%) were the most frequently encountered nosocomial infections caused by Candida species. In addition it was detected that the most frequent causative agent of bloodstream infections was C.parapsilosis (10; 38.5%) and of urinary tract infections was C.albicans (12; 50%). The evaluation of advantages and disadvantages of traditional phenotypic methods [germ tube formation, chlamydospore formation in corn meal agar, growth at 45°C, colony characteristics on CHROMagar Candida medium, carbohydrate assimilation properties detected by API ID 32C (BioMerieux, France) system] and some molecular techniques [polymerase chain reaction (PCR) by using ITS-1, ITS-3 and ITS 4 primers, PCR-Restriction fragment length polymorphism (RFLP), PCRRFLP in which ITS1-ITS4 products cut by Msp I ve Bln I restriction enzymes] for the identification of Candida species revealed that CHROMagar Candida medium combined with API ID 32C kit yielded the same results (100% compatible) as molecular techniques for the species identification of Candida isolates. Since these phenotypic methods were simple and cost effective when compared to molecular techniques, they should be considered in the identification of Candida species.

[The relationship between antibiotic resistance and virulence factors in urinary Enterococcus isolates]. / Üriner Enterokok Izolatlarinin Antibiyotik Direnci ile Virülans Faktörleri Arasindaki Iliski.

Increasing multidrug resistance in nosocomial Enterococcus strains from all over the world recently enhances the need for further investigation of enterococci, especially their virulence factors. There are still many lacking parts about virulence factors of clinical enterococcus isolates. In this study, it was aimed to investigate the antibiotic resistance and the presence of potential virulence factors of 91 Enterococcus strains (59 E.faecalis, 31 E.faecium and 1 E.gallinarum) isolated from urine cultures of inpatients between January 2008-June 2010 in our hospital and also to evaluate whether a correlation existed between antibiotic resistance and potential virulence factors. The genes which encoded virulence factors of enterococci; aggregation substance (AS), enterococcal surface protein (ESP) and hyaluronidase (HYL) (asa1, esp, hyl respectively) were studied by molecular methods and haemolysin production and gelatinase activity were studied by phenotypic methods. Vancomycin resistant strains were checked for the presence of vanA and vanB genes. Eight (25.8%) E.faecium isolates were found glycopeptide resistant. In seven of these isolates resistance type was vanA and in one it was neither vanA nor vanB. High-level gentamicin and high-level streptomycin resistance rates were 74.2% and 61.3% in E.faecium strains and were 22% ve 27.1% in E.faecalis strains, respectively. Beta-lactamase production and linezolid resistance were not detected in any of the strains. E.faecium isolates were more resistant (p< 0.001-0.013) than E.faecalis isolates to all tested antibiotics except tetracycline, minocycline, doxycycline and streptogramin (p< 0.001). hyl gene positivity (p< 0.001) was found higher in E.faecium isolates whereas esp (p= 0.003) and asa1 (p< 0.001) gene positivity, haemolysin production (p=0.014) and gelatinase activity (p= 0.029) were higher in E.faecalis isolates. AS and ESP were the most frequent virulence factors, with the rates of 26.7% and 25.6%, respectively. There were 32 (35.6%) strains without any of the investigated virulence factors. We have also detected that asa1 gene positive E.faecalis isolates were more resistant to ciprofloxacin (p= 0.001), norfloxacin (p= 0.006) and levofloxacin (p= 0.001) than asa1 gene negative isolates; esp gene positive E.faecalis isolates were more resistant to doxycycline (p= 0.043) than esp gene negative isolates and hyl gene positive E.faecium isolates were more resistant to nitrofurantoine (p= 0.011) than hyl gene negative isolates. This was the first clinical sample originated study, investigating the corelation between antibiotic resistance and virulence factors in urinary Enterococcus isolates in Turkey.

[Meningococcemia and meningitis due to Neisseria meningitidis W135 developed in two cases vaccinated with bivalent (A/C) meningococcal vaccine]. / Bivalan (A/C) meningokok asisi ile asilanmis iki olguda Neisseria meningitidis W135'e bagli gelisen meningokoksemi ve menenjit.

Meningococcal infections may develop as episodic or endemic cases particularly among children attending day-care centers, boarding schools or among military personnel. Bivalent (A/C) meningococcal vaccine is applied to all new military stuff since 1993 in Turkey. In this report two cases of meningococcemia and meningitis, developed in two soldiers vaccinated with meningococcal vaccine, were presented. The first case was a 21 years old male patient who was admitted to the emergency service with the complaints of high fever, headache, fatigue and vomiting. He was conscious, cooperative and oriented with normal neurological findings. Maculopapular exanthems were detected at the lower extremities. The patient was hospitalized with the initial diagnosis of sepsis or meningococcemia and empirical treatment was initiated with ceftriaxone and dexamethasone. Cerebrospinal fluid (CSF) examination yielded 10 cells/mm3 (lymphocytes) with normal CSF biochemical parameters. A few hours later skin rashes spread over the body rapidly, the symptoms got worse, confusion, disorientation and disorientation developed, and the patient died due to cardiac and respiratory arrest at the seventh hour of his admission. The second case was also a 21 years old male patient who was admitted to the hospital with the complaints of fever, headache, painful urination, confusion and agitation. He was initially diagnosed as acute bacterial meningitis due to clinical (stiff neck, positive Kernig and Brudzinsky signs) and CSF (8000 cells/mm3; 80% polymorphonuclear leukocytes, increased protein and decreased glucose levels) findings. Empirical antibiotic therapy with ceftriaxone was initiated and continued for 14 days. The patient was discharged with complete cure and no complication was detected in his follow-up visit after two months. The first case had an history of vaccination with bivalent (A/C) meningococcal vaccine three months ago and the second case had been vaccinated one month ago. The bacteria isolated from the blood culture of the first case and the CFS culture of the second case, were identified as Neisseria meningitidis by conventional and API NH system (BioMerieux, France). The isolates were serogrouped as W135 by slide agglutination method (Difco, USA), and both were found to be susceptible to penicillin and ceftriaxone. As far as the last decade's literature and these two cases were considered, it might be concluded that N.meningitidis W135 strains which were not included in the current bivalent meningococcal vaccine, gained endemic potential in Turkey. Since N.meningitidis W135 strains may lead to serious diseases, vaccination of the risk population with the conjugate tetravalent meningococcal vaccine (A/C/Y/W135) should be taken into consideration in Turkey.

Real-time PCR detection of Helicobacter pylori and virulence-associated cagA in nasal polyps and laryngeal disorders.

OBJECTIVE: To identify Helicobacter pylori and major virulence factor, cagA, in patients with laryngeal diseases and nasal polyps. STUDY DESIGN: Cross-sectional study with planned data collection. SETTING: The study was performed on fresh tissue samples from patients with 32 nasal polyps, 29 normal nasal mucosa, and 27 laryngeal diseases presenting to the Otolaryngology-Head and Neck Surgery department of a major military hospital in Istanbul, Turkey. SUBJECTS AND METHODS: Tissue specimens were evaluated by in-house polymerase chain reaction (PCR) and real-time PCR for bacterial DNA and by real-time PCR for cagA. The impact of commercial and in-house DNA extraction methods was also evaluated. RESULTS: H pylori DNA was detected only by real-time PCR in 59.4 percent of nasal polyps, 70.4 percent of nasal mucosa samples, and 58.6 percent of larynx samples. cagA was identified in 78.9, 89.5, and 82.4 percent of positive polyp, nasal mucosa, and larynx samples, respectively. No statistically significant differences were observed between groups. DNA purification methods were equally effective. CONCLUSION: H pylori DNA is present in nasal polyp and larynx tissues as well as normal nasal mucosa, as detected by a sensitive real-time PCR assay. cagA-positive strains dominate in all groups.

[In vitro activity of fosfomycin trometamol in the treatment of Escherichia coli related uncomplicated urinary tract infections]. / Escherichia coli nedenli komplike olmamis üriner sistem enfeksiyonlarinda fosfomisin trometamolün in vitro etkinligi.

Although in certain countries in Europe fosfomycin trometamol (FT) has been used for many years, in Turkey FT has become available in recent years. FT has a broad-spectrum activity against most of gram-positive and gram-negative bacteria. In this study, we aimed to evaluate the effect of FT, a new alternative antimicrobial agent in the treatment of patients with Escherichia coli related uncomplicated lower urinary tract infection (UTI). For this purpose, between May 2007-July 2008, FT susceptibility of 771 nonduplicate E. coli strains, isolated from urine samples of patients with uncomplicated lower UTI (bacteria > or = 10(5) cfu/mL), was determined by disk diffusion method according to Clinical and Laboratory Standarts Institute (CLSI) criteria. Simultaneously, extended-spectrum beta-lactamase (ESBL) detection was performed by double disk synergy test in all isolates. Among all E. coli isolates, FT resistance rate was 0.4% (3/771) and ESBL positivity was 19.5% (150/771). The rates of ESBL producing strains isolated from inpatients and outpatients were 34.1% (70/205) and 14.1% (80/566), respectively, and the difference was found statistically significant (p = 0.0001). Although resistance to FT was not detected in non-ESBL producing E. coli isolates (n = 621), FT resistance rate was 2% (3/150) in ESBL producers. As far as the current literature was concerned this was the largest scale study investigating the activity of FT in Turkey. Resistance to antimicrobials that had been used frequently as therapeutic options for the treatment of E. coli related UTIs, has been increasing. In the present study high susceptibility rates to FT was determined for urinary E. coli isolates. In conclusion, these data suggest that FT may be a good alternative for the treatment of uncomplicated UTIs as a first line antimicrobial agent.

Molecular epidemiology of Blastocystis infections in Turkey.

Blastocystis is a very common unicellular intestinal parasite of ubiquitous occurrence. In order to describe the molecular epidemiology of Blastocystis infections in Turkey, 87 isolates from 69 symptomatic and 18 asymptomatic individuals were sequenced. Sequence data were phylogenetically analyzed and statistically tested against unmodifiable risk factors such as gender and age. Blastocystis-positive males were complaining mainly of gastroenteritis, whereas dyspepsia was the chief complaint among Blastocystis-positive females. Blastocystis sp. subtypes detected in the study included subtypes 1, 2, 3 and 4, subtype 3 being the most predominant (75.9%). No association was detected between Blastocystis sp. subtype and symptoms (p>0.365), or between infection intensity and symptoms (p>0.441). There was a tendency of subtype 2 isolates being more common among older study individuals, and subtype 2 isolates were significantly associated with higher parasite abundance (p=0.017). Compared to data from similar studies, the distribution of Blastocystis sp. isolates in Turkey was found to more or less reflect the one seen in other countries, and it was deduced that subtype 3 is generally by far the most common subtype infecting humans, followed by subtypes 1, 2 and 4.

[Epidemiological features of Candida infections detected in intensive care units and risk factors affecting mortality]. / Yogun bakim ünitelerinde saptanan Candida enfeksiyonlarinin epidemiyolojik özellikleri ve mortaliteye etki eden risk faktörleri.

The aim of this study was to describe the epidemiological features of nosocomial Candida infections in intensive care units and to detect the risk factors which increase the mortality rate. A total of 940 patients hospitalized in ICUs of Gulhane Military Medical Academy Haydarpasa Training Hospital, Istanbul, between January 01 and December 31, 2006 were prospectively enrolled into this study. Candida spp. were isolated from various clinical specimens (blood, urine, respiratory tract, wound) in 48 patients. Of these patients, 50% were male and 50% were female, and the mean age was 63.66 +/- 22.72 (age range: 8-92) years. The mean duration of hospital stay was 36.25 +/- 44.51 (min: 1, max: 90) days. Thirty five Candida infection attacks were observed in 29 of 48 patients. C. albicans was isolated in 18 infections and non-albicans Candida spp. in 17 infections. Nosocomial infection rate due to Candida spp. was 3.22 per 1000 patient-days. The most common Candida infections were bloodstream (42.9%) and urinary tract infections (37.1%). At the time of diagnosis, 89.6% of patients were being hospitalized for more than ten days and 69% of the patients were using three or more wide spectrum antibiotics. Diabetes mellitus and cardiovascular diseases were the most frequently detected co-morbid diseases. The overall mortality rate was 55.2%. Predictors of adverse outcome were diabetes mellitus (p= 0.016), need for mechanical ventilation (p= 0.010) and infection with non-albicans Candida spp. (p= 0.002). In conclusion, Candida infections in the intensive care patients are associated with high mortality. Mortality due to Candida infections could be reduced by defining the risk factors and starting preemptive antifungal treatment to patients who are under risk.

[Antimicrobial resistance profiles of community-acquired uropathogenic Escherichia coli isolates during 2004-2006 in a training hospital in Istanbul]. / Istanbul'da bir arastirma hastanesinde 2004-2006 yillari arasinda izole edilen toplumdan kazanilmis üropatojen Escherichia coli izolatlarinin antibiyotik direnç profilleri.

In order to reveal the antimicrobial resistance profiles against first-line antimicrobial agents in community-acquired acute uncomplicated urinary tract infections (UTIs), resistance patterns were determined for 1664 Escherichia coli strains collected between 2004 and 2006 in GATA Haydarpasa Training Hospital, Istanbul, Turkey. Of the isolates 38.2% were found to be susceptible to all of the tested antimicrobial agents, while the resistance rate to single antibiotic was 13.5%. Highest prevalence of antimicrobial resistance was observed for ampicillin (AMP) (49%), followed by amoxycillin-clavulanic acid (AMC) (34%), sulphamethoxazole/trimethoprim (SXT) (34%) and ciprofloxacin (CIP) (18%). The rate of multidrug resistant isolates was 33.5% and 48.4% of them were co-resistant. Resistance against two antimicrobials was identified in 244, against three antimicrobials in 205, against four antimicrobials in 160, against five antimicrobials in 63 and against six antimicrobials in 23 strains. Most frequent phenotypes indicating resistance against two, three and four antimicrobial agents were AMP/AMC (5.7%), AMP/AMC/SXT (5.4%) and AMP/AMC/cephazolin/SXT (2.6%), respectively. Extended spectrum beta-lactamase (ESBL) activity was detected in 40 (2.4%) of the isolates. Most prominent increases in resistance prevalence during the study period were observed for AMP (from 52% to 63%), AMC (from 33% to 45%) and CIP (from 15% to 22%). These results show that resistance to AMP, AMC and SXT are frequent in community-acquired E. coli strains and empirical initial treatment with these agents will most probably be inappropriate in our region.

Investigation of aminoglycoside modifying enzyme genes in methicillin-resistant staphylococci.

Methicillin-resistant staphylococci may also be resistant to some other antibiotics as well as beta-lactams. In this study, co-existence of resistance to methicillin and aminoglycosides was genetically investigated in staphylococci. A total of 50 staphylococci from in-patients, 17 Staphylococcus aureus and 33 coagulase negative staphylococci (CNS) that contained mecA (gene encoding PBP 2a, an altered penicillin-binding protein) determined by polymerase chain reaction (PCR) were included in the study. Aminoglycoside modifying enzyme (AME) genes were investigated using multiplex-PCR. Aminocyclitol-6'-acetyltransferase-aminocyclitol-2''-phosphotransferase [aac(6')/aph(2'')] gene (encoding bifunctional acetyltransferases/phosphotransferases) was determined in 66% of the isolates, aminocyclitol-4'-adenylytransferase (ant(4')-Ia) gene (encoding phosphotransferases) in 24%, and aminocyclitol-3'-phosphotransferase (aph(3')-IIIa) gene (encoding nucleotidyltransferases) in 8%. Two isolates contained all these three genes. Thirty-six (72%) isolates had at least one of these genes. Three CNS and one S. aureus isolates sensitive to oxacillin had the mecA gene. In conclusion, a high rate of aminoglycoside resistance was determined in methicillin-resistant staphylococci. The aac(6')/aph(2'') was the most frequently detected.

Investigation of erythromycin and tetracycline resistance genes in methicillin-resistant staphylococci.

In this study, erythromycin [erm(A) and erm(C)] and tetracycline [tet(K) and tet(M)] resistance genes were investigated by multiplex polymerase chain reaction (PCR) in a total of 56 methicillin-resistant (mecA+) staphylococcal hospital isolates, 28 of which were determined to be Staphylococcus aureus (MRSA) and the other 28 were coagulase-negative staphylococci (MRCNS). Internal control primers amplifying a specific fragment of 16S rDNA of staphylococci were included in the multiplex PCR protocol to ensure the efficacy of amplification and to determine any PCR inhibition. No resistance genes were detected in 5 of 56 (8.9%) isolates in the study. In the study, tet(K) genes were detected widely (42.9%) in MRCNS, whilst tet(M) genes were detected in MRSA (50.0%). Regarding the erythromycin resistance genes, whilst erm(A) genes were detected in most (71.4%) MRSA isolates, detection rates of erm(C) genes were the same (64.3%) both in MRCNS and MRSA. The resistance rates for tetracycline and erythromycin were 57.1% and 78.6%, respectively, in MRSA isolates. In conclusion, in this study, the multiplex PCR technique including an internal control is shown to be a fast, sensitive, reliable, practical, reproducible and economic technique for the detection of erythromycin and tetracycline resistance in staphylococcal isolates.

[Investigation of methicillin resistance and aminoglycoside modifying enzyme genes in hospital staphylococci by multiplex-polymerase chain reaction]. / Hastane izolati stafilokoklarda metisiliN diRenç ve aminoglikozid modifiye edici enzim genlerinin multipleks-polimeraz zincir reaksiyonu ile aratirilmasi.

This study was conducted to investigate the presence of methicillin and aminoglycoside resistance encoding genes by multiplex-polymerase chain reaction (PCR) and by phenotypic methods in staphylococci isolated from inpatients' clinical specimens. The presence of aac(6')1aph(2"), aph(3')-IIIa and ant(4)-Ia genes encoding aminoglycoside modifying enzymes (AME) and mecA gene encoding methicillin resistance were genotypically investigated. A total of 19 S. aureus and 30 coagulase negative staphylococci (CNS) were tested. Thirty four (69.4%) of the isolates were found to be resistant to oxacillin with disk diffusion test, 33 (97%) of which were found to harbour mecA gene. The correspondance between oxacillin resistance and presence of mecA gene was found to be 100% in S. aureus isolates, while it was 95.7% in CNS. Twenty two (44.9%), 7 (14.3%) and 2 (4.1%) isolates were found to harbour aac(6')/aph(2"), aph(3')-IIIa and ant(4)-/a AME genes, respectively. At least one or more AME genes were detected in 72.7% of mecA positive isolates.

Surgical site infection rates in 16 cities in Turkey: findings of the International Nosocomial Infection Control Consortium (INICC).

BACKGROUND: Surgical site infections (SSIs) are a threat to patient safety; however, there were no available data on SSI rates stratified by surgical procedure (SP) in Turkey. METHODS: Between January 2005 and December 2011, a cohort prospective surveillance study on SSIs was conducted by the International Nosocomial Infection Control Consortium (INICC) in 20 hospitals in 16 Turkish cities. Data from hospitalized patients were registered using the Centers for Disease Control and Prevention (CDC) National Healthcare Safety Network (NHSN) methods and definitions for SSIs. Surgical procedures (SPs) were classified into 22 types according to International Classification of Diseases, Ninth Revision criteria. RESULTS: We recorded 1879 SSIs, associated with 41,563 SPs (4.3%; 95% confidence interval, 4.3-4.7). Among the results, the SSI rate per type of SP compared with rates reported by the INICC and CDC NHSN were 11.9% for ventricular shunt (vs 12.9% vs 5.6%); 5.3% for craniotomy (vs 4.4% vs 2.6%); 4.9% for coronary bypass with chest and donor incision (vs 4.5 vs 2.9); 3.5% for hip prosthesis (vs 2.6% vs 1.3%), and 3.0% for cesarean section (vs 0.7% vs 1.8%). CONCLUSIONS: In most of the 22 types of SP analyzed, our SSI rates were higher than the CDC NHSN rates and similar to the INICC rates. This study advances the knowledge of SSI epidemiology in Turkey, allowing the implementation of targeted interventions.

[Comparison of BACTEC and BacT/ALERT systems in evaluation of blood cultures]. / Kan kültürlerinin degerlendirilmesinde BACTEC ve BacT/ALERT sistemlerinin karsilastirilmasi.

Automatized blood culture systems are used widely due to the characteristics such as lower contamination risks, shorter incubation periods and higher isolation rates. In this study, BACTEC 9050 (Becton Dickinson, Ireland) and BacT/ALERT 120 (Organon Teknika Corp, USA) systems were compared for the evaluation of blood cultures obtained from 222 hospitalized patients in intensive care units between September and December 2003. For this purpose, two blood samples obtained from each arms of patients, were inoculated into BACTEC plus aerobic/F (Becton Dickinson, Ireland) and BacT/ALERT FA/aerobic (BioMeriéux, USA) media, respectively, and incubated. The first bacterial growth time has been recorded for each system and the bacteria were identified by conventional methods. As a result, both of the systems detected bacterial growth in 42 (91.3%) of a total of 46 (20.7%) positive blood samples obtained from 222 patients. The distribution of microorganisms grown in BACTEC and BacT/ALERT were found as follows, respectively; 24 and 20 coagulase negative staphylococci, 8 and 8 Acinetobacter sp., 6 and 10 Escherichia coli, 1 and 1 Staphylococcus aureus, 1 and 1 Pseudomonas sp., 1 and 1 Klebsiella sp., 1 and 1 Candida sp. False positivity and false negativity were not observed in either of the systems. Positive results were obtained in an average time of 20.6 (7.5-35.5) hours and 22.49 (1.5-35.5) hours with BACTEC and BacT/ALERT, respectively. Since there was no statistically significant difference between the systems concerning bacterial isolation rates and periods (p=1.00), it may be concluded that both of these systems would be used in clinical microbiology laboratories for the evaluation of blood cultures.

[Evaluation of intestinal parasites in a period of four years in the coprology laboratory of a training hospital]. / Bir egitim hastanesi koproloji laboratuvarinda geçen dört yillik dönemde saptanan bagirsak parazitlerinin degerlendirilmesi.

Parasitological examination of stool samples of 9867 individuals, submitted from both inpatient and outpatient departments to the Microbiology Department of an 800-bed Training Hospital between 1.1.2003 and 31.12.2006, revealed that 582 individuals (5.9%) were infected with intestinal parasites. Different diagnostic methods were employed in two different periods, and Blastocystis hominis, Giardia intestinalis and Entamoeba histolytica/dispar were found to be the most common parasites, respectively. Patients were predominantly male (67% (n=336) male vs. 33% (n=165) female) and aged between 15-25 years (36%). In addition, 14% (81/582) patients had multiple parasitoses. Despite being labor-intensive, parasitological examination of stool samples with necessary staining methods by experienced staff will surely help determine both the diagnosis and exact prevalence of intestinal parasitic infections in Turkey.

Effects of sevoflurane and/or nitrous oxide on bacterial growth in in vitro culture conditions.

The aim of this study was to determine the role of sevoflurane and/or nitrous oxide on bacterial growth under conditions in vitro similar to those of clinical practice. We assessed these effects on Pseudomonas aeruginosa, Acinetobacter lwoffii, and Staphylococcus aureus growth. Bacterial inoculums were prepared from reference strains in nutritive broth. Airtight chambers were filled with bacterial suspensions. Each strain was studied with and without exposure to sevoflurane and/or nitrous oxide at baseline, after 1 and 3 h. Serial dilutions and agar plates were made and the colonies were counted. P. aeruginosa were grown after exposure to the nitrous oxide alone (2.8 x 10(3) colony-forming units/ml; CFU ml(-1)) after 3 h according to the control (P < 0.05). A. lwoffii were grown after exposure to the nitrous oxide and sevoflurane with nitrous oxide (8.7 x 10(3) and 8.0 x 10(3) CFU ml(-1)) (P < 0.05), respectively. There were no changes in S. aureus growth in controls and anesthesia groups. We conclude that the effects of anesthetic agents on bacterial growth may change owing to the type of anesthetic and microorganism.