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Ozdemir K.,Yuzuncu Yil University | Acar S.,Public Health Institution of Turkey
PLoS ONE | Year: 2014

This study was conducted for typing Salmonella enterica subspecies enterica strains in Turkey using pulsed-field gel electrophoresis (PFGE) and plasmid DNA profile analysis. Fourty-two strains were isolated from clinical samples obtained from unrelated patients with acute diarrhea. The samples were collected from state hospitals and public health laboratories located at seven provinces in different regions of Turkey at different times between 2004 and 2010. The strains were determined to belong to 4 different serovars. The Salmonella enterica strains belonged to the serovars Salmonella Enteritidis (n = 23), Salmonella Infantis (n = 14), Salmonella Munchen (n = 2), and Salmonella Typhi (n = 3). Forty-two Salmonella enterica strains were typed with PFGE methods using XbaI restriction enzyme and plasmid analysis. At the end of typing, 11 different PFGE band profiles were obtained. Four different PFGE profiles (type 1, 4, 9, and 10) were found among serotype S. Enteritidis species, 3 different PFGE profiles (type 3, 5, 6) were found among S. Infantis species, 2 different PFGE profiles were found among S. Typhi species (type 2 and 11), and 2 different PFGE profiles were found among S. Munchen species (type 7, 8). The UPGMA dendrogram was built on the PFGE profiles. In this study, it was determined that 4 strains of 42 Salmonella enterica strains possess no plasmid, while the isolates have 1-3 plasmids ranging from 5.0 to 150 kb and making 12 different plasmid profiles (P1-P12). In this study, we have applied the analysis of the PFGE patterns and used bioinformatics methods to identify both inter and intra serotype relationships of 4 frequently encountered serotypes for the first time in Turkey. © 2014 Ozdemir, Acar.

Yeslyurt M.,Tekirdag State Hospital | Kilic S.,Public Health Institution of Turkey | Celeb B.,Public Health Institution of Turkey | Gul S.,Sorgun State Hospital
Scandinavian Journal of Infectious Diseases | Year: 2013

Tularemia during pregnancy is exceedingly rare and has been reported infrequently in Europe. A review of the literature identified only 3 documented cases. Herein we report 4 tularemia cases occurring early in the second and third trimesters, which were successfully managed without any adverse pregnancy outcomes. © 2013 Informa Healthcare.

Celebi B.,Public Health Institution of Turkey
Türkiye parazitolojii dergisi / Türkiye Parazitoloji Derneǧi = Acta parasitologica Turcica / Turkish Society for Parasitology | Year: 2014

Capillaria hepatica is a nematode with worldwide distribution, which can cause parasitic hepatitis both in animals and humans. A mouse (Apodemus flavicollis), trapped in Giresun Province was diagnosed as having capillariasis due to the characteristic eggs found in its liver. This is the first reported case of mouse capillariasis in this part of the country. Due to the fact that capillariasis is a zoonotic disease, humans might be also infested; therefore, further investigations are needed.

The Clinical and Laboratory Standards Institute (CLSI) Subcommittee on Antifungal Susceptibility Testing has newly introduced species-specific clinical breakpoints (CBPs) for fluconazole and voriconazole. When CBPs can not be determined, wild-type minimal inhibitory concentration (MIC) distributions are detected and epidemiological cutoff values (ECVs) provide valuable means for the detection of emerging resistance. The aim of this study is to determine triazole resistance patterns in Candida species by the recently revised CLSI CBPs. A total of 140 Candida strains isolated from blood cultures of patients with invasive candidiasis hospitalized in various intensive care units in Turkey and sent to our reference laboratory between 2011 - 2012, were included in the study. The isolates were identified by conventional methods, and susceptibility testing was performed against fluconazole, itraconazole and voriconazole, by the 24-h CLSI broth microdilution (BMD) method. Azole resistance rates for all Candida species were determined using the new species-specific CLSI CBPs and ECVs criteria, when appropriate. The species distribution of the isolates were as follows; Cparapsilosis (n= 31 ), C. tropica lis (n= 26 ), Cgtabrata (n= 21), C.albicans (n= 18), Clusitaniae (n= 16), C.kmsei (n= 16), C.kefyr (n= 9), C.guiWermondii (n= 2), and Cdubliniensis (n= 1). According to the newly determined CLSI CBPs for fluconazole and C.albicans, Cparapsilosis, C.tropicalis [susceptible (S), 2 pg/ml; dose-dependent susceptible (SDD), 4 pg/ml; resistant (R), 8 pg/ml], and Cglabrata (SDD, 32 pg/ml; Ite 64 pg/ml) and for voriconazole and C.albicans, Cparapsilosis, Ctropicalis (S, 0.12 pg/ml; SDD, 0.25-0.5 pg/ml; R, 1 (jg/ml), and Ckrusei (S, 0.5 Mg/ml; SDD, 1 pg/ml; R, 2 pg/ml), it was found that three of C.albicans, one of Cparapsilosis and one of Cglabrata isolates were resistant to fluconazole, while two of C.albicans and two of Ctropicalis were resistant to voriconazole. The ECVs of 0.5 pg/ml for voriconazole and C.glabrata were used to differentiate wild-type (MIC ECV) from non-wild-type (MIC > ECV) strains. Five of Cglabrata were non-WT for voriconazole. Due to the lack of CBPs for the less common species, the ECVs for fluconazole, itraconazole and voriconazole, respectively, were used for Clusitaniae (2 pg/ml, 0.5 pg/ml, 0.03 pg/ml), Cguilliermondii (8 pg/ml, 1 pg/ml, 0.25 pg/ml), Cdubliniensis (0.5 pg/ml, 0.25 pg/ml, 0.03 pg/ml), and C.kefyr (1 pg/ml, 0.015 pg/ml) to categorize isolates of these species as wild- And non-wild-type. When the ECVs were used for fluconazole, one each of Clusitaniae, Cdubliniensis and C.kefyr, for voriconazole, three of Clusitaniae and one of Ckefyr were detected as non-wild-type. Overall, a total of five Candida species were resistant to fluconazole and four to voriconazole and among these species one each of Cparapsilosis, Ctropicalis, C.glabrata, Clusitaniae, C.kefyr and three of C.albicans exhibited cross-resistance at least against two azoles. It was concluded that, the strains identified as resistant and non-wild-type in this in vitro study should be supported by molecular and in vivo studies for the determination of their clinical validity.

Kilic S.,Public Health Institution of Turkey | Celebi B.,Public Health Institution of Turkey | Acar B.,Public Health Institution of Turkey | Atas M.,Public Health Laboratory
Scandinavian Journal of Infectious Diseases | Year: 2013

Background: Tularemia is an infection caused by Francisella tularensis, which has a wide distribution in the northern hemisphere and diverse clinical manifestations. For decades, the drug of choice for the treatment of tularemia has been streptomycin, with tetracycline and chloramphenicol being used as alternatives. The purpose of the present study was to determine the in vitro antimicrobial susceptibility of a large panel of geographically diverse F. tularensis isolates from Turkey against traditional and newer antimicrobial agents. Methods: The antibiotic susceptibilities of 250 F. tularensis strains were examined using the Epsilometer test for 9 antimicrobial agents. Each isolate was identified by conventional and molecular techniques. Results: All the strains were confirmed biochemically and using a combination of species-and subspecies-specific polymerase chain reaction (PCR) assays to be F. tularensis subsp. holarctica. One isolate was assigned to F. tularensis subsp. holarctica biovar japonica based on erythromycin susceptibility, an ability to ferment glycerol, and the nucleotide sequence of the region of difference 1 (RD1). All strains were susceptible to aminoglycosides (streptomycin and gentamicin), tetracyclines (tetracycline and doxycycline), chloramphenicol, 2 fluoroquinolones (ciprofloxacin and levofloxacin), and rifampicin. In addition, all isolates except 1 had a minimal inhibitory concentration (MIC) for erythromycin of > 256 μg/ml. Conclusions: Since the fluoroquinolones showed the lowest MIC values and have advantages such as excellent bioavailability and activity, availability of oral formulations, and lower toxicities, they represent candidate therapeutic options in the first-line treatment of tularemia. To the best of our knowledge, this is the first report of the presence of F. tularensis subsp. holarctica biovar japonica outside Japan. © 2013 Informa Healthcare.

Coskun K.A.,Gaziosmanpaşa University | Ozgur A.,Gaziosmanpaşa University | Otaug B.,Cumhuriyet University | Mungan M.,Public Health Institution of Turkey | Tutar Y.,Cumhuriyet University
Protein and Peptide Letters | Year: 2013

Toxoplasma gondii is ubiquitous obligate intracellular parasite and is one of the most important pathogen for humans and animals. In humans, T. gondii has two life forms: tachyzoites and bradyzoites. Tachyzoites form of T. gondii can cause acute infection, and it is called toxoplasmosis. The development of latent bradyzoites from rapidly growing tachyzoites has been linked to cellular and environmental stresses which are associated with heat shock proteins (Hsps). Hsps play a protective role against stressors. Hsp40 is an important member of Hsp family and T. gondii has 36 predicted Hsp40 family members. Therefore, we studied the cloning and biochemical characterization of the T. gondii RH strain Hsp40 protein-Gok1. Hsp40 prevents protein aggregation and induce refolding. Consequently, Hsp40s may play essential roles in the mechanisms of bradyzoite development and survival in the host organism. Hsp40-Gok1 functional and structural properties may facilitate drug design and protein targeting against toxoplasmosis. © 2013 Bentham Science Publishers.

Ata N.,Konya Training and Research Hospital HacI Saban | Kilic S.,Public Health Institution of Turkey | Ovet G.,Konya Training and Research Hospital HacI Saban | Alatas N.,Konya Training and Research Hospital HacI Saban | Celebi B.,Public Health Institution of Turkey
Infection | Year: 2013

Tularemia is a zoonotic infection caused by Francisella tularensis with a worldwide distribution and diverse clinical manifestations. Although F. tularensis has been recognized as a human pathogen for a century, there are few reports regarding the occurrence of tularemia in pregnant women and its effect on the fetus; only seven cases have been reported in the literature. In view of the sparse literature, it is not clear whether tularemia increases the risk of adverse pregnancy outcomes. In this paper we review tularemia infection during pregnancy, its complications and management. In addition, we present a case of tularemia that occurred in the first trimester of pregnancy and resulted in third-trimester intrauterine fetal death, highlighting the consequences of tularemia in pregnancy and the importance of early detection and treatment. © 2013 Springer-Verlag Berlin Heidelberg.

Kilic S.,Public Health Institution of Turkey | Celebi B.,Public Health Institution of Turkey | Yesilyurt M.,Tekirdag State Hospital
Diagnostic Microbiology and Infectious Disease | Year: 2012

Tularemia is an infection caused by Francisella tularensis with a worldwide distribution in the northern hemisphere and diverse clinical manifestations. Serology plays an important role in the diagnosis of tularemia. A commercially available immunochromatographic assay (ICA) for the serologic diagnosis of tularemia (VIRapid Tularemia, Vircell, Granada, Spain) was evaluated, and the performance was compared with that of the current standard, the microagglutination test (MA). A panel of 221 sera from 109 cases of tularemia was tested as well as 236 sera from normal individuals or individuals with other infectious or autoimmune diseases. The ICA demonstrated 91.5% (κ = 0.91) agreement with the reference method (MA) and gave an overall sensitivity of 99.3% and a specificity of 94.6%. No cross-reactivity was observed in the ICA with serum samples from normal individuals and patients with autoimmune diseases and bacterial, viral, and parasitic infections, although 4 of 50 patients with brucellosis demonstrated positive results in the ICA. The performance of ICA was simple, and it requires no specialized equipment. The ease of use and significantly high sensitivity and specificity of ICA make it a good choice for diagnostic testing and a valuable field test to support a presumptive diagnosis of tularemia in remote areas. © 2012 Elsevier Inc.

Celebi B.,Public Health Institution of Turkey | Kilic S.,Public Health Institution of Turkey | Yeslyurt M.,Tekirdag State Hospital | Acar B.,Tekirdag State Hospital
Mikrobiyoloji Bulteni | Year: 2014

Tularemia is a rare zoonotic infection, however, considerations of tularemia as a biological weapon and several recent major epidemics have caused renewed interest in this disease. Laboratory diagnosis of tularemia is done in the presence of appropriate epidemiological data, by the demonstration of specific antibodies in the serum samples obtained with 1 -2 week intervals following the development of symptoms. It is an a posteriori analysis with limited use for prompt diagnosis of the patient during the early symptomatic phase and deliberate release of biological agents. Limitations in both culture and serology have led to substantial research in the development of new diagnostic techniques. Several PCR methods for tularemia have been developed, both for conventional and real-time polymerase chain reaction (rtPCR). However, PCR methods are hard to be deployed in remote endemic areas that lack sufficient infrastructure. Recently a "Toolbox" which includes all instruments, equipments and solutions [DNA4U® Bacteria Genomic DNA Isolation Kit, CubeCyder® (Personal Thermal Cycler), PCR4U® Bioterrorism Agents Detection Kit, electrophoresis tank, power supply, ready-agarose gel and electrophoresis buffer] necessary for conventional PCR, was developed for the identification of bioterrorism agents in the field. In this study we aimed to evaluate the efficacy of a ready-to-use commercial PCR kit (Nanobiz, Ankara, Turkey) targeting the tul4 gene, for the diagnosis of tularemia and to compare the results with an in-house conventional PCR and a rtPCR test. We applied the assay to a collection of four F.tularensis standard strains, 15 field isolates (from humans, animals, water), 13 non-Francisella strains which are phylogenetically related to F.tularensis and a total of 60 lymph node aspirates obtained from suspected tularemia cases. Compared to the in-house PCR method used in our laboratory, the sensitivity, specificity, positive and negative predictive values of Nanobiz PCR Toolbox assay were found to be 100%. The lowest detection limit of this method was determined as 100 genomic equivalent per PCR reaction mix. The new PCR kit is a rapid and accurate alternative to the conventional PCR methods since the toolbox includes all of the required chemicals, accessories and equipments. This ready-to-use PCR assay was appraised to be a valuable diagnostic tool for the detection of F.tularensis in the outbreak settings particularly in remote areas with limited resources.

Ulu Kilic A.,Erciyes University | Kilic S.,Public Health Institution of Turkey | Celebi B.,Public Health Institution of Turkey | Sencan I.,Diskapi Yildirim Beyazit Education and Research Hospital
Mikrobiyoloji Bulteni | Year: 2013

Francisella tularensis is the etiological agent of tularemia which is a zoonosis of the northern hemisphere. For decades, streptomycin was considered the drug of choice, despite possible side effects, and vestibular toxicity in particular. Alternatives are tetracylines and chloramphenicol which are bacteriostatic agents that are associated with a considerable risk of relapse. The aim of the present study was to assess the in vitro susceptibility of F.tularensis subsp. holarctica biovar II strains to tigecycline, a member of a new class of glycylcyclines. Fourteen F.tularensis strains isolated from patients in Central Anatolia region of Turkey were examined. Minimum inhibitory concentration (MIC) values of tigecycline, doxycycline, streptomycin, gentamicin, and ciprofloxacin were determined using the E-test method on glucosecysteine blood agar plates. Interpretation of results was made according to CLSI clinical breakpoints. All strains were susceptible to the antibiotics traditionally used to treat tularemia. Tigecycline showed good in vitro activity to all the isolates (MIC range: 0.094-0.38 mg/L). In this study, tigecycline was more active than doxycycline against F.tularensis subsp. holarctica strains, according to MIC50 (0.19 mg/L) and MIC90 (0.25 mg/L) values. Doxycycline (MIC90: 0.38 mg/L) showed good in vitro activity against all the isolates and MIC values interpreted according to the CLSI criteria for potential bioterrorism agents, have shown ranges below the breakpoint for sensitivity determination (S ≤ 4 mg/L). Ciprofloxacin had the lowest MIC50 and MIC90 values. In case the other antibiotics can not be used or intravenous the rapy is required, tigecycline may be an important therapeutic alternative agent. However, confinement of tigecycline in the treatment of multi-drug resistant bacterial infections, its parenteral way of administration and overall cost were considered as the major limitations of tigecycline in tularemia treatment.

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