Investigation of the relationship between virulence factors and antibiotic resistance of Enterococci isolates.

The aim of this study was to determine the relationship between  aggregation factor (asa1), enterococcal surface protein (esp), cytolysin (cyl), gelatinase (gelE), hyaluronidase (hyl) virulence factors and antibiotic resistance in Enterococci. VITEK 2 ID system was used to identify the isolates and determine their antibiotic susceptibility. Virulence genes were investigated by polymerase chain reaction. Of the 93 isolates, 62 (66 %) were Enterococcus faecium, 31 (44 %) were Enterococcus faecalis (E. faecialis ). E. faecium isolates were more resistant to ampicillin, ciprofloxacin, linezolid, teicoplanin and vancomycin than E. faecalis. High-level gentamycin rate were higher in E. faecium than E. faecalis (p <0.05). The most prevalant virulence genes were esp (60.9 %) and asa1 (25 %) followed by gelE (22.8 %), cyl (16.3 %) and hyl (8.7 %). Asa1, cyl, gelE genes positivity were higer in E. faecalis than E. faecium. Hyl positivity was higher in E. faecalis than E. faecium isolates. Ampicillin resistance was higher in gelE positive E. faecalis than gelE negative E. faecalis (p <0.05). Ciprofloxacin resistance was higher in gelE negative E. faecalis than gelE positive E. faecalis (p <0.05). Asa, cyl, hyl, gelE positive E. faecium isolates were more susceptible to teicoplanin than the isolates that did not have these genes (p <0.05). Cyl, asa, gelE positive E. faecalis isolates were more susceptible to vancomycin than cyl, asa, gelE negative E. faecalis isoates (p <0.05). Hyl positive E. faecium isolates were more susceptible to vancomycin than hyl negative E. faecium isolates (p <0.05). E. faecalis isolates that have virulence genes were more susceptible to vancomycin (p <0.05). The resistance to antibiotics in E. faecalis should be a concern for the treatment of infectious disease.

Genotypic analysis of S segment of Crimean-Congo hemorrhagic fever virus in Turkey.

The Crimean-Congo hemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne virus, whose viral RNA consists of S, L, and M segments. The virus is migrating through the ticks with animals and migratory birds and the geographical distribution can be investigated based on genetic analysis. To better understand the connection between the seropositivity and the mortality rate, the key factor could be the temporal and spatial analysis of the different strains. In this study, serum samples (n = 26) were selected from CCHFV RNA-positive patients and subjected to sequence analysis of the gene regions encoding the S segments. According to the neighbor-joining analysis, the obtained partial sequences were linked to the European strain. The strains were closely related to Turkey-Kelkit06, Turkey 200310849 viruses, and viruses from Russia and Kosovo. The comparison with previously analyzed isolates from the GenBank showed 95%-99% sequence similarity. The isolates in phylogenetic branches were divided into two groups. AST, platelet, and APTT levels were found significantly higher in Group 2 compared to Group 1. Nucleotide differences can be prognostic factor in CCHF disease. Increasing CCHF cases not originating from local isolates were circulating strains imported from different neighboring countries of Turkey. The results show new evidence to the emerging threat of the CCHF disease.

Investigation of plasmid mediated AmpC beta-lactamases in Escherichia coli and Klebsiella pneumoniae isolates by phenotypic and genotypic.

OBJECTIVE: To investigate the susceptibility and specificity of the phenotypic methods to determine plasmidmediated AmpC. METHODS: The cross-sectional study was conducted at Duzce University Faculty of Medicine, Microbiology Laboratory from January 2015 to June 2016, and comprised Escherichia coli and Klebsiella pneumonia isolates intermediate susceptible or resistant to cefoxitine. Combined disk diffusion test, double disc synergy test, agar gradient test and polymerase chain reaction were used to detect plasmid-mediated AmpC. RESULTS: Of the 2024 E. coli samples, 44(2.17%), and of the 792 K. pneumoniae samples, 16(2%) were included. Combined disk diffusion test had susceptibility of 68% and specificity of 50%; double disc synergy test 24% and 82%; and agar gradient test 40% and 68%. Of the isolates positively detected by polymerase chain reaction method, more than one gene region positivity was detected in 15(25%) isolates. CONCLUSION: All three phenotypic methods were found to be insufficient to detect plasmid-mediated AmpC positivity.

Association of eNOS gene 4a/4b VNTR and T786C polymorphism with Crimean-Congo hemorrhagic fever.

The most common viral hemorrhagic fever is Crimean-Congo hemorrhagic fever (CCHF). Endothelial nitric oxide synthase (eNOS) gene polymorphisms have been linked to both hemorrhagic fevers and viral diseases. The study's goal is to evaluate if the eNOS gene 4a/4b and T786C polymorphisms are related to CCHF. The study included 54 CCHF RNA-positive patients and 60 control subjects. The Bosphore CCHF virus Quantification Kit v1 was used to obtain CCHF RNA, and the Magnesia 16 isolation device was used to isolate DNA (Anatolia Gene works, Turkey). Polymerase chain reaction and restriction fragment length polymorphism were used to genotype the samples. The frequency of the eNOS 4a/4a, 4a/4b, and 4 b/4b genotypes in patients and the control was 6.6% versus 1.7%, 37.0% versus 43.3%, and 57.4% versus 55%, respectively. 4a: 24.07% of patients and 23.33% of controls; and 4 b: 75.92% of patients and 76.66% of controls. The frequency of the eNOS-786 T/C, T/T, T/C, and C/C genotypes in patients and the control group was 35.2% versus 68.3%; 51.9% versus 26.73%; and 13.0% versus 5.0%, respectively. The allele and genotype frequencies of the eNOS T786C variant differ statistically between patients and the control (p < 0.05). The eNOS T786C variant could be a genetic determinant for susceptibility to CCHF. To our knowledge, this is the first study to figure out the association between eNOS gene T786C polymorphisms and CCHF disease.

Neonatal Thrombocytopenia and the Role of the Platelet Mass Index in Platelet Transfusion in the Neonatal Intensive Care Unit

Background: Neonatal thrombocytopenia is a common hematological abnormality that occurs in 20­35% of all newborns in the neonatal intensive care unit. Platelet transfusion is the only known treatment; however, it is the critical point to identify neonates who are really at risk of bleeding and benefit from platelet transfusion as it also has various potential harmful effects. Aims: To investigate the prevalence and risk factors of neonatal thrombocytopenia and its relationship to intraventricular hemorrhage in the neonatal intensive care unit and to determine whether the use of platelet mass index-based criteria could reduce the rate of platelet transfusion. Study Design: Retrospective cohort study. Methods: This study was conducted in the neonatal intensive care unit of a tertiary university hospital. The medical records of neonates in the neonatal intensive care unit with platelet counts <150×109/L between January 2013 and July 2016 were analyzed. Results: During the study period, 2,667 patients were admitted to the neonatal intensive care unit, and 395 (14%) had thrombocytopenia during hospitalization. The rate of intraventricular hemorrhage was 7.3%. Multiple logistic regression analysis showed that although lower platelet counts were associated with a higher intraventricular hemorrhage rate, the effects of respiratory distress syndrome, sepsis, and patent ductus arteriosus were more prominent than the degree of thrombocytopenia. Thirty patients (7%) received platelet transfusion, and these patients showed a significantly higher mortality rate than their non-platelet transfusion counterparts (p<0.001). In addition, it was found that the use of platelet mass index-based criteria for platelet transfusion in our patients would reduce the rate of platelet transfusion by 9.5% (2/21). Conclusion: Neonatal thrombocytopenia is usually mild and often resolves without treatment. As platelet transfusion is associated with an increased mortality rate, its risks and benefits should be weighed carefully. The use of platelet mass index-based criteria may reduce platelet transfusion rates in the neonatal intensive care unit, but additional data from prospective studies are required.

ß-lactamase genes in carbapenem resistance Acinetobacter baumannii isolates from a Turkish university hospital.

INTRODUCTION: The spread of Acinetobacter baumannii, resistant to most of the available antimicrobial agents, is a serious health problem. The high rate of carbapenem resistance among Acinetobacter baumannii isolates is considered as a threat to public health. In this study, we aimed to determine the antibiotic resistance and related genes in carbapenem-resistant Acinetobacter baumannii isolates. METHODOLOGY: Ninety six isolates of A. baumannii were included. Antimicrobial susceptibility was performed by Phoenix Automated System and disk diffusion method. Carbapenem resistane was characterized by scrneeing of resistance genes such as blaTEM, blaSHV, blaCTX-M1-2, blaPER, blaVEB, blaKPC, blaGES, blaNDM, blaVIM, blaIMP and blaOXA23-24-51-58 using multiplex polymerase chain reaction. RESULTS: Resistance for the levofloxacin, gentamicin, amikacin, and tigecycline were determined as 96.9%, 93.7%, 72.9% and 45.8% respectively. Colistin was the only susceptible antibiotic against all clinical isolates. All isolates were defined as multidrug resistance and of these, 31.2% were extensively drug-resistant (sensitive only to colistin). BlaOXA-51-  and blaOXA-23 genes were detected in 100% strains while blaTEM was found in only 2% strains. There was no amplification for the blaSHV, blaCTX-M1-2, blaPER, blaVEB, blaKPC, blaGES blaNDM, blaVIM, blaIMP and blaOXA24-58 genes. CONCLUSIONS: The high frequency of blaOXA-23 and low frequency of blaTEM gene was observed that indicate prevalence of a variety of A. baumannii strains. The rates of resistance genes vary from region to region. Studies are required for the prevention and control of A. baumannii infection and to formulate the strategies of antibiotic usage.