Search

outcomes of stapled and hand-sewn Roux-en-Y intestinal anastomoses in childhood. Materials and Methods: At a university hospital, the records of the children who underwent roux-en-Y anastomosis between December 2007 and December 2014 were reviewed. The data were compared according to the roux-en-Y anastomosis technique used (stapled versus hand-sewn). Results: A total of 52 patients had undergone roux-en-Y anastomosis. All had biliary atresia or choledochal cyst. Thirty-one of the patients were diagnosed with biliary atresia and 21 with choledochal cysts. Staple anastomosis technique was used in 16 of the patients with biliary atresia and 9 of the patients with choledochal cyst. Both in biliary atresia and choledochal cyst cases; operations with stapled anastomosis were significantly shorter than the ones with hand-sewn anastomosis. Among the biliary atresia cases, post-operative oral feeding was initiated significantly earlier in the stapled group, with its lower risks of post-operative cholangitis and longer hospital stay than 7 days. Conclusion: This is the first study in children, confirming the time-saving advantage of stapled anastomosis over hand-sewn, during roux-en-Y anastomosis for biliary atresia and choledochal cyst; along with the safety of stapler use, including the neonates. Stapled anastomosis yields lower complication rates, faster function gain with earlier feeding and hospital discharge.

Aim Non-operative treatment approach is another method used in the treatment of uncomplicated appendicitis, in which the infection in the appendix is suppressed and treated with antibiotics. Our study aims to investigate the clinical outcomes and the risk factors for recurrence in our pediatric patients with suspected uncomplicated appendicitis, who underwent non-operative treatment. Methods The medical data of the patients who underwent non-operative treatment with the diagnosis of suspected uncomplicated appendicitis between January 2016 and January 2021 in a tertiary pediatric surgery center were analyzed. Demographic data, treatment process, and clinical results of the patients were recorded. Statistical evaluation was made by comparing the two groups with and without recurrence after non-operative treatment. Results The median age of 41 patients whose data were evaluated was 13 (6-17) years. Eight patients (19.5%) had appendicolith. The median duration of IV antibiotic treatment was 4 (3-7) days, and the patients' abdominal tenderness disappeared in a median of 2 (1-4) days. Recurrence developed in 8 (19.5%) patients after a median of 7 (1-14) months after non-operative treatment. It was found that the time to the disappearance of abdominal tenderness was statistically longer in the group that developed recurrence than that in the group that did not (p=0.01). Conclusion Our study revealed that appendicolith was not a risk factor for the development of recurrence. The time to the disappearance of abdominal tenderness may be useful for detecting patients at a higher risk of recurrence.

Çalışan kişiye uyacak şekilde iş, ekipman ve işyeri tasarlama bilimi "ergonomi" olarak adlandırılır. Ergonomi, endüstriyel şirketler tarafından işçilerin görevlerini ve çalışma alanlarını tasarlamak için yaygın olarak kullanılır. Ergonominin amacı, insan-makine etkileşimini geliştirerek sistemlerin performansını artırmaktır. Bu bağlamda kullanıcı arayüzü tasarlanarak, bunun yanısıra ilgili sistemler geliştirilerek görev ve kullanıcı ile daha uyumlu hale getirilebilir. Ergonomi; antropometri, biyomekanik, genel mühendislik, fizyoloji, psikoloji ve fizik gibi çeşitli disiplinleri içerir. Burada değinilmesi gereken önemli konulardan biri; başta boyun, sırt, bilek, kol, omuz ve bacaklar olmak üzere bazı vücut bölgelerinin riske maruz kalma düzeyini belirleyen risk değerlendirmesidir. Olumsuz etkilerini önlemek için risk düzeyini belirlemek önemlidir. Bu amaçla, risk seviyelerini belirlemek için Hızlı Tüm Vücut Değerlendirmesi (REBA), Hızlı Üst Ekstremite Değerlendirmesi (RULA) ve Hızlı Maruziyet Değerlendirme (HMD) gibi çeşitli yöntemler kullanılır. Ancak risk seviyesi sektörlere göre değişmektedir. İnşaat sektörü en riskli sektörlerden biridir ve önemine göre bu çalışmada ele alınmıştır. İnşaat endüstrisindeki bazı yaygın süreçler seçilmiştir ve üç ergonomi risk değerlendirme yöntemi; bir Türk inşaat firmasındaki riskleri ortaya çıkarmak için REBA, RULA ve HMD uygulanmıştır. İlgili sonuçlar karşılaştırmalı olarak sunulmuş ve yorumlanmıştır.

Bu çalışmada, yükseköğretim ek ders ücret yönetim süreçlerinde yaşanan sorunların çözümü için geliştirilen bir bilişim sisteminin kullanılabilirliğinin belirlenmesine odaklanılmıştır. Bu amaçla araştırmacılar tarafından bir bilişim sistemi geliştirilmiştir. Bilişim sistemi uygulamaya konulmadan önce bir öntest yapılarak kullanıcıların ek ders süreçleriyle ilgili karşılaştıkları sorunlar tespit edilmiş ve memnuniyet düzeyleri ölçülmüştür. Sistem devreye alındıktan iki yıl sonra bir sontest uygulanarak mevcut sorunların azalıp azalmadığı belirlenmiş ve bilişim sisteminin kullanılabilirlik düzeyi ölçülmüştür. Elde edilen bulgulardan bilişim sisteminin kullanılabilirlik skorunun kabul edilebilir bir düzeyde olduğu tespit edilmiştir. Sistemin kullanımıyla birlikte sorunların azaldığı ve memnuniyet düzeyinin arttığı görülmüştür. Kullanılabilirlik düzeyi yüksek ürün ve hizmetlerin özellikleri açısından geliştirilen bilişim sistemi değerlendirilmiş ve gelecek araştırmalar için önerilerde bulunulmuştur.

Q fever is a zoonosis caused by Coxiella burnetii. In this report, a case of chronic Q fever endocarditis with pancytopenia and hypergammaglobulinemia mimicking a lymphoproliferative disease was presented. A 39-years-old male living in Çatalca and whose family is engaged in animal husbandry admitted with the complaints of weakness and fatigue. The patient had aortic valve replacement 29 years ago and had aortic valve re-replacement, and ascending aorta grafting because of endocarditis three years ago. It was revealed that the second operation of the patient was due to possible infective endocarditis, but no definitive agent could be identified. He was evaluated for massive hepatosplenomegaly, pancytopenia, hypergammaglobulinemia, presence of M-spike and elevated β-2 microglobulin levels and was referred to our hematology clinic with a preliminary diagnosis of lymphoproliferative disease. Lymphoplasmacytic lymphoma was excluded with the result of bone marrow biopsy and he was referred to our clinic for the investigation of possible infectious etiologies. We detected hepatosplenomegaly and finger clubbing. His blood analyses were normal except for the following: leukocyte count 3800/μl, platelet count 148000/μl, gamma globulin 5.9 gr/dl, rheumatoid factor (RF) and antinuclear antibody (ANA) positivity. Chronic Q fever endocarditis was suspected and C.burnetii Phase I IgG test was found positive in 1/132071 titers. Although transesophageal echocardiography showed no lesion of endocarditis, positron emission tomography/computed tomography revealed increased fluorodeoxyglucose uptake around the prosthetic heart valve and graft. The patient was diagnosed as having Q fever endocarditis and graft infection. He refused hospitalization and was started on hydroxychloroquine and doxycycline treatment. The patient stopped taking these antibiotics by himself seven days after the diagnosis. He was admitted with a headache to another hospital and operated for an intracranial hemorrhage and died shortly after. Apart from unfamiliarity, wide range of clinical presentations of disease could also lead to delayed diagnosis. Among patients with chronic Q fever, continuous bacteremia and antigenic stimulus causes inflammatory syndrome with hepatosplenomegaly, hypergammaglobulinemia and, presence of autoantibodies which leads to misdiagnoses of rheumatologic, autoimmune or hematologic diseases Chronic Q fever should be investigated in patients with known valvulopathy and chronic hepatomegaly or splenomegaly, pancytopenia, hypergammaglobulinemia, and unexplained autoantibody positivity.

Shortly after the first detection of human immundeficiency virus (HIV) infection in USA in 1981, the number of cases have increased gradually from all around the world. Turkey’s high capacity for tourism and the unique geographic location extending between Europe and Asia, provides convenience for the passage of individuals across the countries and sexually transmitted infections including HIV, as well. According to the official data of the Ministry of Health; there are 25809 HIV positive and 1958 AIDS cases as of November 30, 2020, after the epidemic started in 1985 in Turkey. Despite the decrease in the number of newly detected HIV cases as a result of serious measures taken for the transmission of infection worldwide, the increase in the number of cases still continues in our country. Shortening the reporting period and starting treatment as soon as possible in the diagnosis of infection is critical for the control of the epidemic. For this purpose, Centers for Disease Control and Prevention (CDC) published a new test algorithm in 2010, which suggested the use of the Geenius™ HIV ½ supplemental assay test instead of western blot tests, which have been used for many years to verify HIV screening test positivity. In this study, we aimed to report the experience of the National HIV-Acquiner Immundeficiency Syndrome (AIDS) and Viral Hepatitis Reference Laboratories of Turkey in the first year of transition to the new HIV algorithm and to evaluate the diagnostic performance of Geenius™ HIV ½ and line immunassay (LIA) s. A total of 2090 anti-HIV positive patient sera sent to National HIV-AIDS and Viral Hepatitis Reference Laboratories of Turkey, Ankara for HIV confirmation were included in the study. All samples were retested with a fourth-generation enzyme linked immunosorbent assay (ELISA) test (VIDAS® HIV-1/2 Duo Ultra assay, BioMerieux, France) followed by the confirmatory tests; Geenius™ HIV 1/2 confirmatory assay (BioRad, Redmond, WA) and Line-immunoassay (INNO-LIA HIV ½ Score, Fujirebio, Belgium). Indeterminate/negative test results or discrepancies between the confirmatory tests were resolved with HIV-1 RNA reverse transcriptase polymerase chain reaction (RT-PCR) (artus HI Virus-1 RT-PCR, Qiagen, Germany) test and in-house HIV-2 RNA and proviral DNA PCR. The sensitivity, specificity, and the agreement of the each assay were compared. Cohen’s Kappa analysis was used for the evaluation of the agreement between the tests. According to the new algorithm which recommended Geenius™ test besides HIV-1 RNA test, 1707 (81.7%) HIV-1 positive samples were identified. Of these samples; 95.9% and 95.02% were identified as HIV-1 positive by GeeniusTM and INNO-LIA, respectively. However, 2.5% of the positive samples were negative with Geenius™ and 3.5% with INNO-LIA. One and a half percentage (1.5%) of these samples were detected with Geenius™ and 1.4% with INNO-LIA as indeterminant. When all the positive samples determined with ELISA were evaluated; it was detected that,1.3% were indeterminate by Geenius™ test and 2.4% by the INNO-LIA test. When the INNO-LIA test was regarded as the gold standard method; sensitivity, specificity, positive predictive and negative predictive values of the Geenius™ test were as follows; 99.7%, 96.1%, 98.9%, and 99.1%. The agreement between INNO-LIA and Geenius™ tests was found to be 98.95% (κ= 0.969; very good). When the Geenius™ and HIV-1 PCR tests were evaluated together for the confirmation; the sensitivities of Geenius™ and INNO-LIA tests were 99.8% and 98.3%, specificities were 89.8% and 85.3%, respectively. Slight positive bands were detected in the gp36 or gp140 bands, the HIV-2 specific envelope proteins, were detected in seven samples, However, the positivity disappeared after the dilution of the samples and it was accepted as false positivite reaction due to the absence of HIV-2 RNA and proviral DNA in these samples. In conclusion; we concluded that Geenius™ and INNO-LIA tests have a perfect agreement in HIV diagnosis and due to the rapid and reliable results provided for the HIV test protocol, Geenius™ test can be used safely as an alternative to the immunoblot tests. HIV-1 RNA testing must be performed in all HIV confirmation centers in order to detect acute HIV cases in the fast and early period which are the main reason for the updates in HIV diagnosis.

Objective: The aim of this study was to evaluate the results of examination for intestinal parasites in fecal samples sent to our laboratory by obtaining from patients applied to the hospital because of various complaints.

Amaç: İnfluenza rutin tanısında yüksek duyarlılık ve özgüllüğü nedeniyle real-time PCR testi tercih edilmektedir. Ancak PCR testlerinin yüksek maliyet ve deneyimli personel gereksinimi, laboratuvar ortamına ve cihazlarına ihtiyaç duyulması, numune transferi ve hekime sonucun ulaşmasında geçen süre gibi nedenlerle hasta başı kullanılan hızlı influenza tanı testlerinin kullanımı gündeme gelmiştir. Bu çalışmanın amacı, influenza enfeksiyonu tanısında kullanılan 3 farklı hızlı tanı testinin gerçek zamanlı RT-PCR yöntemi ile karşılaştırılması ve tanısal performansının belirlenmesidir.Yöntem: Çalışmamıza 2017-2018 influenza sezonunda laboratuvarımıza gönderilen toplam 209 solunum yolu örneği dahil edilmiştir. Örnekler Humasis Influenza antigen card plus (Kore), SD Biosensor Standard-Q Influenza A/B (Kore) ve SD Biosensor Standard-F Influenza A/B FIA (Kore) hızlı antijen tanı kitleri ile üretici firmaların önerileri doğrultusunda test edilmiştir.Bulgular: Çalışmamızda Humasis kiti ile İnfluenza A için duyarlılık %80,99, İnfluenza B için %68,66 hesaplanmıştır. SD Biosensor Standard Q Influenza A/B ve SD Biosensor Standard F Influenza A/B FIA kiti ile her iki virüs için de duyarlılık %70’in altında bulunmuştur. Humasis kitinin duyarlılığı diğer kitlere oranla daha yüksek saptanır- ken özgüllük tüm kitlerde %90’ın üzerinde belirlenmiştir.Sonuç: Bu çalışma ile hızlı antijen testlerinin, influenza virüs aktivitesinin yoğun olduğu influenza sezonu dönemi veya salgın durumlarında influenza virüs enfeksiyonu tanısında tarama testi veya destekleyici test olarak kullanılabileceği sonucuna varılmıştır., Aim: Due to its high sensitivity and specificity, real-time PCR testing is preferred for routine diagnosis. However, for PCR tests, the use of rapid influenza diagnostic tests at bedside due to reasons such as high cost, experienced personnel, laboratory conditions and equipment requirement, sample transfer and test time has come to the fore. Purpose of the study was to compare three different diagnostic tests used in the diagnosis of influenza infection with the real-time RT-PCR method and to determine the diagnostic performance.Methods: A total of 209 respiratory tract samples sent to our laboratory during the 2017-2018 influenza season were included in our study. The samples were tested with Humasis Influenza antigen card plus, SD Biosensor Standard-Q Influenza A/B, and SD Biosensor Standard-F Influenza A/B FIA rapid antigen diagnostic kits according to the recommendations of the manufacturers.Results: In our study, susceptibility was calculated 80.99% for influenza A and 68.66% for influenza B with Humasis kit. With the SD Biosensor Standard Q Influenza A/B and the SD Biosensor Standard F Influenza A/B FIA kit, the sensitivity for both viruses was below 70%. While the sensitivity of the humasis kit was higher than the other kits, the specificity was higher than 90% in all kits.Conclusion: n this study, it was concluded that rapid antigen tests can be used as a screening test or supportive test for the diagnosis of influenza virus infection during the influenza season period or epidemic conditions during which influenza virus activity is intense.

It has been reported that direct identification from blood culture bottles with positive signals and reporting the results to the clinics earlier has positive effects on mortality and morbidity. Extraction methods especially using detergents are used for the direct identification from the bottles which give positive signal. For this purpose, in-house methods developed based on the usage of saponin are widely available in the literature. In this study, it was aimed to develop a simple, easy-to-apply and reliable protocol for identifying the agent directly from the blood culture bottle that gives positive signal with the use of detergent Tween® 80, and to study the obtained protocol in clinical samples in a routine microbiology laboratory and to evaluate the results. The study was carried out in two stages, the experimental stage where the method was developed and the clinical stage where the method was applied. In the experimental stage, blood culture bottles were created with standard strains and isolates previously diagnosed with the 16S rRNA method. 10% solution of Tween® 80 was prepared with distilled water. 1 ml sample was transferred from the bottle that gave positive signal to the microcentrifuge tube, 100 µl of 10% solution of Tween® 80 was added, vortexed for 10 seconds and then incubated for 5 minutes at room temperature. The tubes were centrifuged for 5 min at 14.000 rpm, the supernatant was discarded and the pellet was washed with 1 ml of distilled water and centrifuged at 14.000 rpm for 5 minutes in three times. Samples taken from the pellets were rubbed on the slide and dried on air. Firstly, 1 µl of 70% formic acid, then 1 µl, of matrix solution was added and it was used after drying. In the second stage of the study, the method was applied to the 502 vials giving positive signal in the Microbiology Laboratory of Ankara University Faculty of Medicine Ibni Sina Hospital between 17 April 2018-31 August 2018 and the results were compared with the subculture results. The results obtained at the end of extraction in the experimental stage were compared with the subculture results and no statistical difference was found. In 383 (82.9%) bottles among 462 (92.1%) bottles with monomicrobial positive cultures, compatible results with the subculture results were obtained. Of the microorganisms correctly identified, 350 (91.3%) were bacteria and 33 (8.7%) were fungi. On the other hand, 216 (56.4%) of the bacteria were gram positive and 134 (34.9%) of them were gram negative bacteria. At least one microorganism was correctly identified in 19 (47.5%) of 40 (7.9%) bottles with polymicrobial blood cultures. Their distribution was gram negative (n= 10) and gram positive (n= 8) and yeast (n= 1). No microorganisms were identified in six bottles with polymicrobial cultures. According to the results, we believe that this in-house method developed using Tween® 80 will be a routinely applicable method for blood culture bottles that give positive signal in microbiology laboratories and it will contribute to the early diagnosis.

Francisella tularensis is a gram-negative, coccobasillus, facultative intracellular bacteria and causes a zoonotic disease, tularemia in humans. F.tularensis has four subspecies, which have different virulences for humans as F.tularensis subsp. tularensis, F.tularensis subsp. holarctica, F.tularensis subsp. mediasiatica and F.tularensis subsp. novicida. F.tularensis subsp. tularensis is the most virulent subspecies and mortality rate is high in human cases. F.tularensis subsp. holarctica, which has been reported in our country to date, has lower virulence than that of subsp. tularensis, and causes rare lethality among untreated patients. According to the erythromycin resistance and the properties of glucose-glycerol fermentation, F.tularensis subsp. holarctica has three biovar as biovar I, biovar II and biovar japonica. F.tularensis subsp. mediasiatica has been reported only in a few central asian countries and its virulence is similar to the F.tularensis subsp. holarctica F.tularensis subsp. novicida is avirulent for immunocompetent individuals but has been observed to cause infection in immunocompromised individuals. The aim of this study was to determine the F.tularensis subspecies in 259 F.tularensis strains isolated from clinical specimens, drinking water and a rodent sample and 517 F.tularensis PCR-positive DNA isolated from clinical specimens between years 2009 and 2014. Conventional PCR was performed using primers specific for the RD1 (Region Difference) region of F.tularensis. Subspecies were differentiated depending on the difference in PCR amplification product size. In our study, F.tularensis subsp. holarctica was detected in 764 samples yielding 922 base pair (bp) amplification product. The DNA samples obtained from one water and 11 lymph aspirates were determined as F.tularensis subsp. holarctica biovar japonica. The DNA sequence analysis of the amplification product of the RD1 region of the isolate from water sample was determined. The 1136 bp nucleotide sequence obtained from the DNA sequence analysis was 100% similar to F.tularensis subsp. holarctica biovar japonica (FCS075 strain-accesion number AF469618) when compared with GenBank data. The whole genome sequence of this isolate was also determined and recorded to GenBank with accesion number CP007148. None of the samples used in our study belonged to other sub-species. F.tularensis subsp. holarctica biovar japonica positive 11 lymph aspirate samples were sent to our center from Ankara (n= 1), Kayseri (n= 1) and Afyon (n= 9) provinces. The results of the current study revealed that F.tularensis subsp. holarctica biovar japonica caused a tularemia outbreak in a village in Afyon province at first time and it was observed sporadically in two other different provinces.

Rickettsia species are gram negative, small pleomorphic coccobacilli, obligate intracellular bacteria. The majority of these bacteria are transported by the ticks. Rickettsia slovaca and Rickettsia raoultii are among the Rickettsia species in the spotted fever group and carried by Dermacentor species ticks, that cause tick-borne lymphadenopathy (Tickborne Lymphadenopathy-TIBOLA or Dermacentor-borne necrotic erythema and lymphadenopathy- DEBONEL). Rickettsia species are obligate intracellular bacteria and they can be cultivated in cell cultures. The chance of isolation of Rickettsia species from clinical and tick specimens has been increased by the shell-vial centrifugation cell culture method. The aim of this study was to isolate R.slovaca from two Dermacentor marginatus species ticks which were detected in humans by the use of shell-vial centrifugation cell culture method using VERO cell. Before proceeding to the culture stage, an algorithm including description, disinfection, dissection of ticks and methods of hemolymph, homogenization, DNA extraction and real-time PCR was performed. Iodine-alcohol disinfection was performed following identification of the ticks with the use of a stereo microscope. Ticks hemolymph were obtained with extremity dissection of ticks and indirect fluorescence antibody (IFA) test was performed with Rickettsia positive sera. Ticks identified as containing Rickettsia like bacteria in hemolymph were homogenized and DNA extraction was performed from homogenate of ticks. By real-time PCR, using Rickettsia genus-specific PanR8 primers and probes, PCR positive Rickettsia spp. were determined among the ticks. Vero cells that have formed monolayer in vials were used for Rickettsia culture. Homogenized tick samples which were positive for Rickettsia in hemolymph and real-time PCR methods were inoculated to cell culture vials. Suspended bacteria in the inoculum were allowed to approach the cells by the shell-vial centrifugation method. Cells were scraped after five days of incubation in 5% CO2 at 36°C. An aliquot of the determined cell suspension was fixed to the slides and then IFA was performed using Rickettsia positive sera. In fluorescence microscopy examination, adhered and proliferated bacteria were observed on Vero cells. This cell suspension was again inoculated into the Vero cell cultures to increase bacterial replication and taken for 5-7 days of incubation. DNA was extracted from the suspension of the cultivated bacteria. Conventional PCR of citrate synthase (gltA) and outer membrane protein A (ompA) gene regions were used to identify Rickettsia species. DNA sequence analysis of PCR amplification products were determined. DNA sequence results were compared to Genbank data and found that the gltA sequence was 100%, similar to R.slovaca with accession number AY129301.1 and the ompA sequence was 100%, similar to R.slovaca with accession number KF791242.1. In addition, the two strains isolated in phylogenetic analysis were found to be R.slovaca. As a result, R.slovaca isolation from of D.marginatus type ticks has been reported for the first time in our country by the cell culture method.

Pertussis is an important cause of morbidity and mortality in infants under two months of age and these high risk babies are dependent on maternally derived antibodies until completion of their first immunization series. This study aimed to evaluate the vaccine response of late preterm and term newborns as well as their mothers who underwent combined tetanus-diphtheria toxoid and acellular pertussis (Tdap) vaccination during pregnancy.A total of 70 pregnant women were administered Tdap vaccine (Boostrix®, GSK) between 27 and 33 gestational weeks of pregnancy. The IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in maternal blood before vaccination and in both maternal and umbilical cord blood after vaccination were evaluated using the in-house ELISA method. The geometric mean concentrations (GMC) and placental transfer ratios of antibodies were measured.Participants' with a mean age of 29.59 ± 4.70 years received Tdap vaccine at an average 28.6 ± 1.31 gestational weeks. Average pre and post vaccination levels of anti-PT IgG GMCs and anti-FHA IgG GMCs were 8.01 IU/ml vs 39.48 IU/ml (p = 0.001) and 122.24 IU/ml vs 183.97 IU/ml (p 0.001), respectively. The anti-PT and anti-FHA IgG GMCs of cord blood after vaccination was 25.15 IU/ml and 118.77 IU/ml, respectively (p 0.001 and p = 0.064). Placental transfer ratios of anti-PT ve anti-FHA IgG antibodies were detected as 0.65 and 0.62, respectively.Immunization of pregnant women with Tdap at the third trimester results in high maternal and infant antibody levels. Maternal immunization during each pregnancy seems to be the best strategy in revealing the highest maternal and infant antibodies and in narrowing the gap between birth and immune system maturation in infants. Pregnant women in our country should also get the Tdap vaccine during pregnancy especially in the early third trimester.

Objective: Antibiotic resistance is one of the most significant problems of human-animal and environmental ecosystems. It is crucial to establish integrated surveillance systems and monitor resistance for the management of antibiotic resistance. Standardization of antibiotic resistance data obtained from various disciplines is the critical point in enhancing the data quality. To realize this objective, a common antibiotic susceptibility testing training program was prepared and performed for professionals from various disciplines to standardize the resistance data to be obtained from the human, animal, and environmental sectors in our country. Method: A total of 48 individuals participated in a five-day training program in three terms. In each period, four small groups, each consisting of four people from a group of different professions, were generated. Participants were trained on quality control, phenotypic tests, genotypic tests, and the use of resistance data in antibiotic susceptibility testing. Pre-test and post-tests were applied to the participants. Results: Individuals with a postgraduate degree who studied antibiotic susceptibility testing in the fields of medicine, veterinary medicine, pharmacy, food, and environment participated in the training. The average number of correct answers in the pre-test and post-test increased from 4.8 to 10.5 in April, from 4 to 9 in June and from 3.4 to 8.5 in September. They studied phenotypic and genotypic tests in the supplied isolates under the supervision of the educators. Conclusion: We presume that dissemination of the training at graduate and postgraduate levels will also enable the One-Health approach to become widespread. In addition, worldwide application of similar trainings will help standardization of resistance data, as well as one health approach.

Background Serological methods have great importance for the detection of Treponema pallidum antibodies in syphilis diagnosis. The goal of the present study is to evaluate various commercially available screening assays in comparison with the FTA-abs test. Methods A total of 363 serum samples were enrolled in the study. Following routine testing including RPR and TPHA tests, each sample was tested by treponemal immunoassays (Chorus Syphilis Screen Recombinant, Architect Syphilis TP, Syphilis Virclia Monotest, Siemens Advia Centaur Syphilis, Euroimmun Treponema pallidum Screen ELISA, Vircell Syphilis ELISA IgG + IgM, SD Bioline Syphilis). The result obtained from each test was compared with the confirmatory FTA-abs test. Kappa (κ) coefficients were used to compare the concordance of the tests. Results When the various tests were evaluated in comparison with the FTA-abs test, the sensitivity, specificity and percent agreement of each test were as follows: Architect Syphilis TP, 92.3%, 94.5%, 92.8%; Chorus Syphilis Screen Recombinant, 87.9%, 91.2%, 88.7%; Syphilis Virclia Monotest, 80.5%, 97.8%, 84.9%; Siemens Advia Centaur Syphilis, 87.5%, 89%, 87.9%; Euroimmun Treponema pallidum Screen ELISA, 87.5%, 85.7%, 87.1%; Vircell Syphilis ELISA IgG + IgM, 73.2%, 62.6%, 70.5%; TPHA, 89%, 63.7%, 82.6%; SD Bioline Syphilis, 58.1%, 94.5%, 67.2%; RPR test, 57.7%, 57.1%, 57.6%. Conclusion The results of the present study show that Treponema pallidum specific immunoassays with a performance similar or better than TPHA test generally performed well with the confirmatory FTA-abs test and may be an alternative for screening total antibodies in syphilis infection.

Background/aim: Limited data on syphilis coinfection in human immunodeficiency virus (HIV) positive cases exist in Turkey. Our aim is to investigate syphilis coinfection and to evaluate the compatibility of the screening Architect Syphilis Tp ELISA with the fluorescent treponemal antibody absorption (FTA-abs) confirmation test in HIV positive cases. Materials and methods: Totally 519 HIV positive patients were included in the study. Enzyme linked fluorescent assay (ELFA) was used as a screening test and positive samples were confirmed by line immunassay (LIA). In order to discriminate acute HIV infection and false ELISA positivity, HIV-1 RNA PCR was performed in ELFA positive and LIA negative samples. Architect Syphilis TP ELISA was used for the detection of total antibodies against Treponema pallidum in HIV positive patients. Positive results were confirmed by the FTA-abs test. Results: Out of 519 HIV-1 positive patients, IgG and IgM positivity, and only IgG positivity was detected as 1.9% and 11.4% in all the samples, respectively. A total of 79 (15.2%) sera were positive with Architect Syphilis Tp ELISA test and 69 (13.3%) were positive with FTA-abs test. Statistically significant, almost perfect agreement was found between Architect Syphilis Tp ELISA and FTA-abs tests (kappa = 0.921 and P < 0.001). Conclusion: Implementation of syphilis and HIV screening tests together among risk groups is considered to be appropriate.

Although a significant decrease has been reported in the incidence of diphteria in many regions of the world following the routine diphtheria immunization programs, the emergence of new cases indicated that toxigenic strains are still circulating in the community. Diphtheria vaccine does not provide protection against asymptomatic carriage and colonization of non-toxigenic Corynebacterium diphtheriae. It is a known fact that invasive infections may arise from non-toxigenic C.diphtheriae strains that the non-toxigenic strains can become toxigenic strains leading to diphteria. It is also known that there is a risk of diphteria outbreaks due to decreased antitoxin level and inadequate adult immunization programs. In our country, there is no routine surveillance of toxigenic and non-toxigenic C.diphtheriae. In the present study we aimed to investigate the presence of C.diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis in children presenting with the symptoms of upper respiratory tract infections that might be confused with those moderate diphteria, in order to highlight the requirement of microbiological surveillance and to create awareness about these microorganisms among public health experts, microbiologists and clinicians. Throat swab specimens were obtained from children who were admitted to the pediatric outpatient clinics, in Dr. Sami Ulus Obstectrics, Children Health and Diseases Educational and Research Hospital, with upper respiratory tract infections between 1 February 2016-22 March 2016. The specimens were inoculated in 5% sheep blood agar plates. The plates that were incubated in appropriate conditions, were evaluated for Group A beta hemolytic streptococcocci. Subsequently, culture plates were sent to the Public Health Institution of Turkey, National Respiratory Pathologens Reference Laboratories for the investigation of the presence of C.diphtheriae, C.ulcerans and C.pseudotuberculosis. The growth in each plate were collected with a sterile swab and inoculated in tryptic soy broth. Following 2 hours of incubation at 37oC, subcultures were inoculated in cystine-tellurite-blood agar (CTBA) and 5% sheep-blood agar plates; after an overnight incubation tellurite-reducing colonies were inoculated in Tinsdale agar plates. The suspected colonies with positive cystinase activity were identified by conventional methods and also with Coryne API (Biomerieux, France) systems. Toxicity tests (ELEK, PCR) were performed to investigate whether the C.diphtheriae strains were producing toxins. A total of 500 patients were involved in the study. Of these 260 (52%) were girls and 240 (48%) were boys with a mean age of 76 (range, 21-213) months. All patients except one were fully vaccinated with boosters. Most common presenting symptoms of the patients were fever (19.8%), sore throat (52.6%), cough (49.2%), tonsillar hyperemia (97.6%), presence of crypt (24.6%), and membrane over tonsils (1%). Group A beta-hemolytic streptococcocci were detected in the throat swab cultures of 66 (%13.2) patients. Genotypically toxin negative C.diphtheriae biovar gravis was identified in the throat swab cultures of 3 patients (2 girls and 1 boy). The tonsils were hyperemic and hypertrophic in all the patients with C.diphtheriae biovar gravis. C.ulcerans and C.pseudotuberculosis were detected in none of the patients. It is considered that similar regular cross-sectional studies or routine screening programs are expected to raise awareness about this forgotten microorganism both epidemiologically and microbiologically.

Purpose Toxoplasma gondii is an intracellular parasite that can affect all vertebrae and is the causative agent of toxoplasmosis. At present, the United States CDC (Centers for Disease Control and Prevention) recognizes this infection as a neglected disease. Toxoplasma gondii infection profiles exhibit differences because of the different regional and climatic responses to these parasites in Turkey, and these protozoan infections are notably common in this country. In this study, we attempted to obtain the whole-genome sequence of T. gondii using next-generation sequencing technology. Methods Toxoplasma gondii isolates were isolated from an infant with congenital toxoplasmosis by Ekmen et al. (1974) in Ankara, Turkey. Whole-genome sequencing (WGS) was performed using the Illumina HiSeq 2500 and HiSeq SBS Kit v2. A T. gondii library was created on this device in the initial stage. After the completion of the library phase, sequence analysis was begun with a next-generation sequencing device. The resulting fragments were combined using paired-end (PE) reading and converted into a single DNA fragment. Bioinformatic analysis was performed using the Geneious 2.1. program. Results In our study, WGS was successfully performed on T. gondii. The T. gondii whole-genome sequence has a coverage value of 50x, a size of 61,5763 Mb and a GC ratio of 52.6%. Data from this sequence were submitted to the National Center for Biotechnology Information (NCBI) GenBank ( www.ncbi.nlm.nih.gov ) database under the name Toxoplasma gondii TR01 (TG_TR01). The accession number of the genome obtained in this study is WOEV00000000.1. The biological sample access number is SAMN13338796. The genome of the T. gondii strain obtained in this study was compared with the reference genome, and 8312 CDSs (coding sequences), 183 tRNAs, 294 rRNAs and 8789 genes were identified. Among the 8312 CDSs, 4284 encoded hypothetical proteins (hypothetical protein CDSs/proteins of unknown function). The entire genome sequence of T gondii TR01 was compared with that of Toxoplasma gondii ME49. The results of this comparison demonstrate that the analyzed genome was 99,98% similar to the reference genome. The accession numbers of 14 chromosomes belonging to the genome sequences of T. gondii TR01 (TG_TR01) are CM019722.1, CM019723.1, CM019724.1, CM019725.1, CM019726.1, CM019727.1, CM019728.1, CM019729.1, CM019730.1, CM019731.1, CM019732.1, CM019733.1, CM019734.1, and CM019735.1. Conclusion In this study, a whole-genome sequences of T. gondii was conducted for the first time in Turkey. The analyzed strain was named T. gondii TR01. The data obtained from this study may contribute to a better understanding of T. gondii. T. gondii is an important pathogen with an unusual population structure. Although T. gondii is highly zoonotic and has a complicated life cycle, some strains of this parasite have exhibited high genetic sequence similarity, and our study supports this knowlegde. The characterization of this strain may be very useful for the scientific community of our country and may help to establish a foundation for further research investigating the genome of T. gondii.

Brucella melitensis and Brucella abortus account for almost all cases of brucellosis in Turkish population. We developed a fourplex quantitative real-time PCR (qPCR) assay for the electrophoresis-free, rapid and cost-effective differentiation of B. abortus and B. melitensis from the other Brucella spp. The 4-plex species differentiation assay was combined with a qPCR assay targeting 17 different single nucleotide polymorphism (SNP) loci in Brucella genomes. This combination resulted in a 21 Variable Genome Loci (21-VGL) qPCR assay for high resolution genotyping of B. abortus and B. melitensis. A total of 486 Brucella was analyzed using the qPCR assay to create a 21-VGL profile database. The database contained the profiles of 55 B. abortus, 352 B. melitensis, 3 B. ceti, 6 B. neotomae, 7 B. ovis, 6 B. pinnipedialis, 44 B. suis and 13 B. canis strains. The 21-VGL Brucella genotyping clearly distinguished B. abortus, B. melitensis, B. neotomae and B. ovis. The 21-VGL approach could not distinguish B. pinnipedialis from B. ceti and some B. suis genotypes from B. canis. The results revealed that more than 99% of the Brucella isolates in Turkey were B. melitensis and 21-VGL genotyping can be reduced to 8-VGL B. melitensis genotyping without any loss of genotyping resolution. To our knowledge, we introduced the fastest and the lowest-cost B. abortus and B. melitensis genotyping and species differentiation methodology in the literature.

In this study, we described the largest analysis to date conducted with VIDAS® HIV Duo Ultra assay. Additionally, we analyzed the diagnostic performance and cutoff values (TV) of HIV Duo Ultra assay and total cost analysis for HIV testing. Of 11,642 enzyme-linked immunosorbent assay (ELISA)-positive samples referred to our center for confirmation, 2000 were positive with HIV Duo Ultra, and of these, 87% were HIV-1 positive and 0.6% were HIV-1 indeterminate with the confirmatory test. Overall, the false-positivity rate was 1.75% for HIV Duo Ultra assay. The sensitivity and specificity were 100% and 99.1%, respectively, when the TV was set at the recommended cutoff value. Even increasing the cutoff value four times, sensitivity and specificity remained high, pointing out that a TV of 0.99 is highly indicative of HIV positivity. Retesting samples with HIV Duo Ultra assay decreased 80% of the confirmatory tests, revealing a significant decrease of 78% in the total costs and reporting time.

WOS: 000365461000023 PubMed ID: 26583383 Francisella tularensis DNA extractions and isolates from the environment and humans were genetically characterized to elucidate environmental sources that cause human tularemia in Turkey. Extensive genetic diversity consistent with genotypes from human outbreaks was identified in environmental samples and confirmed water as a source of human tularemia in Turkey. United States Department of Homeland Security (DHS) Cowden Endowment in Microbiology at Northern Arizona University

Meningococcal conjunctivitis is a rare but important infection since it can lead to severe complications and can threaten public health. It may emerge in two forms, either primary or secondary type which is developed after a systemic infection. Accurate diagnosis of primary meningococcal conjunctivitis is very important in addition to ocular complications which can result in loss of vision, the condition can also lead to severe complications like systemic meningococcal disease. However, the lack of specific symptoms which can distinguish meningococcal conjunctivitis from other forms of bacterial conjunctivitis, initiation of empiric antibiotic therapy without performing culture and nonaccurate differentiation of Neisseria gonorrhoeae and Neisseria meningitidis with commercial kits/systems used in laboratories cause problematic situations. This report describes a case of primary unilateral conjunctivitis in a 14-month-old girl caused by non-groupable N.meningitidis that was resolved without sequelae following treatment. A pre-healthy 14-month-old girl was brought to the pediatric emergency department with redness, crusts and discharge in the left eye that had begun two days earlier. Ocular examination revealed hyperemia and purulent discharge in the left conjunctiva. Purulent conjunctivitis was diagnosed. A conjunctival swab specimen was taken for culture, and the patient was started on topical netilmicin (4x1), topical fusidic acid (2x1) and artificial tears. Microscopic examination of the conjunctival swab revealed polymorphonuclear leukocytes and no visible bacteria. Catalase and oxidase positive, gram-negative diplococci grew purely in culture. The first Gram stain preparation was evaluated again after the growth and small numbers of gram-negative diplococci were observed. The cultivated bacteria were identified as N.meningitidis using MALDI-TOF MS (Bruker Daltonics, Germany), but as N.gonorrhoeae with BBL Crystal N/H (Neisseria/Haemophilus) (BD Diagnostic Systems, MD) identification system. The isolate was identified as N.meningitidis by polymerase chain reaction method. The isolate was sent to the Public Health Institution of Turkey for confirmation and serotyping. It was confirmed as non-groupable N.meningitidis. This is the first report of conjunctivitis caused by non-groupable N.meningitidis from Turkey. We wish to emphasize the importance of Gram staining and differentiation of the species by automatized systems in diagnosis, netilmicin may be one of the options for empiric treatment and in terms of public health the most appropriate approach may be evaluation of the severity of conjunctivitis and causative serogroup which depends on case-based approach.

Background: Brucellosis is a zoonotic disease caused by Brucella species. Although brucellosis is considered as an occupational disease in adults, recently it has become an infectious disease affecting all age groups, including children. Molecular epidemiological studies are crucial for control and treatment of disease in children. Objectives: This study aimed at identifying Brucella species, to detect antibiotic susceptibilities and define transmission dynamics between the Brucella isolates in children. Methods: A total of 77 Brucella isolates were identified by conventional and polymerase chain reaction methods. Anti - biotic susceptibilities were investigated by E - test strips. The isolates were genotyped by using multiple locus variable number tandem repeat analysis (MLVA) (MLVA - 16 Orsay), including 8 mini - satellite (panel 1) and 8 microsatellite (panel 2A and 2B) markers. Results: The mean age was 9.14 ± 3.4 years. All patients had been consuming unpasteurized milk. All isolates were Brucella melitensis biovar 3. Only 2 isolates were resistant to ceftriaxone, while the other isolates were susceptible to other antimicrobial agents. The MLVA - 16 typing revealed 42 MLVA profiles. Eighteen profiles included 2 or more isolates, indicating a clustering rate of 66.7%. Twenty - four isolates showed a unique profile. Single locus, double locus, and 3 locus variants were detected in 32, 26, and 15 isolates, respectively. Bruce 30, Bruce 16, Bruce 9, Bruce 7, and Bruce 4 were highly discriminatory loci, respectively. All strains were defined as genotype 122, according to MLVA - 11, and genotype 43 according to MLVA - 8, and were in the Eastern Mediterranean genotype. Conclusions: High clustering rate revealed that brucellosis among the children mainly resulted from common sources. Controlling animal movements and avoiding contaminated milk products have an importance to interrupt spread of brucellosis in children.

Orf is a zoonotic infectious disease caused by parapoxvirus. Orf lesions are typically seen on the hand, but they have rarely been reported on the nose. Herein, the authors report a rare patient of an orf lesion on the nose of a 52-year-old man after the Muslim celebration of the feast of the sacrifice. The lesion spontaneously recovered 8 weeks after the initial appearance and showed no evidence of recurrence after 1 year of follow-up. Orf virus infections may occur more often after the celebration of the feast of the sacrifice in Muslim countries.

OBJECTIVE This study was conducted to identify the seroprevalance of diseases which are zoonotic and responsible from abortion such as toxoplasmosis, listeriosis and brucellosis in Holstein crossbred cattle of different age and sex in Adana province, between 2008 April-September. METHODS For this purpose, blood serum samples were collected from 132 cattle and analyzed for Toxoplasma gondii, Listeria monocytogenes and Brucella abortus antibodies. T. gondii, L. monocytogenes and Brucella abortus antibodies were determined by the standard Sabin- Feldman Dye Test (SFDT), Osebold method and Microtube Agglutination Test (MAT) respectively, from the blood serum samples. RESULTS 132 serum tested 56.06% samples of T. gondii, 40.9% and 3.03% of L. monocytogenes and defined the B.abortus antibodies were found to be seropositive terms. There were no statistically significant difference between seropositive T. gondii, L. monocytogenes and B.abortus antibodies among age groups (p>0.05). CONCLUSION In this study, for the first time in cattle in the region of Adana serological methods revealed the presence L. monocytogenes, toxoplasmosis and listeriosis were higher than brucellosis seropositivity. Moreover, the prevalence of these diseases in the same animal at the highest rate was determined for T. gondii and L. monocytogenes.

Recently reports of cervical tuberculous lymphadenitis and oropharyngeal tularemia which are the most common infectious causes of granulomatous lymphadenitis, have been significantly increased in Turkey. The differentiation of cervical tuberculous lymphadenitis and oropharyngeal tularemia is usually confusing on the basis of clinical and histopathological findings. Thus, in tularemia endemic areas, the patients are more commonly evaluated in terms of tularemia lymphadenitis leaving tuberculosis out. The aim of this study was to investigate the presence of Mycobacterium tuberculosis in cervical lymph node aspirates, obtained from tularemia suspected cases. A total of 105 oropharyngeal tularemia-suspected cases which were found negative for Francisella tularensis by bacteriological (culture), molecular (PCR) and serological (microagglutination) methods, were included in the study. The samples had been previously studied at National Tularemia Reference Laboratory, Turkish Public Health Institution, between 2009-2011. The study samples were evaluated in terms of M.tuberculosis by culture and real-time PCR (rtPCR) methods in the National Tuberculosis Reference Laboratory. Both Lowenstein-Jensen (LJ) medium and liquid-based MGIT (BD, USA) automated culture system were used for mycobacterial culture. Samples that yielded mycobacterial growth were identified as M.tuberculosis by immunochromotographic test (BD, USA). The lymph node aspirates of 65 patients who were F.tularensis PCR negative but antibody positive, were used as the control group. As a result, M.tuberculosis was found to be positive in 9 (8.6%) of 105 tularemia-negative lymph node aspirates, sent to our laboratory from different geographic regions for the investigation of tularemia. Six of the M.tuberculosis positive cases were male and the age range of the patients was 26-85 years. The presence of M.tuberculosis was detected only by culture in two samples, only by rtPCR in five samples and both by culture and rtPCR in two samples. M.tuberculosis was not identified in the control group specimens. Three of the samples which revealed tuberculosis, were from the tularemia endemic areas. In conclusion, the data of this preliminary study indicated that tuberculous lymphadenitis should be kept in mind in suspected tularemia cases and those patients should also be investigated simultaneously for the presence of tuberculous lymphadenitis.

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, CubeCycler® (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.

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.

Anthrax, caused by the bacterium Bacillus anthracis, is one of the oldest documented infectious diseases in both livestock and humans. We aimed to evaluate clinical findings and risk factors of patients with cutaneous anthrax infection and report anti-lethal factor (LF) IgG and anti-protective antigen (PA) IgG titers in the serologic diagnosis of disease.In this study, serum samples of 18 cutaneous anthrax patients were collected and anti-LF IgG and anti-PA IgG titers were measured by enzyme-linked immunosorbent assay (ELISA).Twelve (67%) males and 6 (33%) females, with a mean age of 36.06 ± 16.58 years were included in the study. Risk factors identified in the patient population studied were slaughtering (28%), flaying (56%), chopping meat (67%), burying diseased animal corpses (17%) and milking (6%) livestock. Black eschar formation (94%), pruritus (78%) and painful lymphadenopathy (61%) were first three common clinical signs and symptoms, respectively. Fourteen (78%) patients produced a positive IgG response against PA, 11 (61%) patients produced against LF. Three (17%) patients had no response to either antigen.A detailed history of contact with sick animals or animal products along with clinical findings should be taken at the first step for the diagnosis of cutaneous anthrax infection. Serologic detection of anti-LF IgG and anti-PA IgG with ELISA may be useful auxillary method for establishing the diagnosis.

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.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.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 of256 μg/ml.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.

The aim of the present study was to assess the antimicrobial susceptibility of Brucella melitensis isolates to rifampicin (RIF) depending on time and regional differences. A total of 94 human Brucella isolates collected in an 8-year period from the beginning of 2002 to the end of 2009 throughout Turkey were investigated. The isolates were identified at species and biovar levels by conventional methods, and minimum inhibitory concentrations (MIC) of RIF was determined by using the E test method. All isolates were identified as B. melitensis (93 isolates, biovar 3; 1, biovar 1), and MIC(50) and MIC(90) values of RIF were 1 and 1.5 μg/ml, respectively (MIC range, 0.25-1.5 μg/ml). All isolates were sensitive to RIF except 2 isolates, which had intermediate susceptibility to RIF. These findings indicated that B. melitensis biovar 3 may be the most frequently agent responsible for human brucellosis in Turkey. None of the isolates in our region was resistant to RIF.

Objective We aimed to attract our colleagues’ attention to the oropharyngeal tularemia, which has been an infrequently diagnosed disease in otolaryngology clinics also becoming widespread in Turkey. Methods A retrospective review of database of 3 patients was performed to find patients with Tularemia. The medical files, treatment charts, serological and PCR findings and radiological (magnetic resonance imaging), histopathological records of these patients were reviewed. Results The study group consisted of 3 oropharyngeal tularemia patients. All of them presented with tonsillopharyngitis and cervical lymphadenitis and all were resistant to beta lactam or cephalosporin group antibiotics. Two patients had magnetic resonance imaging findings. All patients had positive serological tests. Strongly positive serological tests led to the diagnosis of tularemia. All patients had positive PCR and direct fluorescent antibody (DFA) test. All patients responded to systemic streptomycin treatment. Conclusion In the daily practice of an otolaryngologist, it is not usual to diagnose a patient with oropharyngeal tularemia. Tularemia should be considered in the differential diagnosis of massive adenotonsillar enlargement and extensive necrotic cervical lymphadenopathy not responding to beta lactam antibiotics as a result of the rising number of tularemia outbreaks outside the classic endemic areas.

Purpose: Brucellosis is a worldwide zoonotic disease and still constitutes a major public health problem. In the study we claimed to identify Brucella species from clinical samples of patients with active brucellosis from Van region of Eastern Anatolia and to determine in vitro antimicrobial susceptibilities of these strains to commonly used anti-Brucella agents and a possible new alternative tigecycline. Materials and Methods: A total of 56 Brucella isolates were enrolled the study and the identification of the isolates were based on conventional methods. In vitro activities of an-timicrobials were evaluated by the E test method. Results: All isolates were identified as B. melitensis. MIC90 values of doxycycline, strepto-mycin, rifampin, trimethoprim-sulfamethoxazole and tigecycline were 0.064 mg/L, 1 mg/L, 2 mg/L, 0.125 mg/L and 0.094 mg/L, respectively. Tigecycline had low MIC50 and MIC90 values against all B. melitensis strains; the highest MIC observed was 0.25 ?g/mL. Conclusion: Our data suggest that tigecycline can be a therapeutic alternative option for the treatment of brucellosis. © Ivyspring International Publisher.

Antimicrobial therapy of Brucella spp. infection is difficult because there are relatively few effective treatment regimens, and single-agent therapy has generally been considered inadequate due to unacceptably high relapse rates. tigecycline, the first in a new class of antimicrobials, the glycylcyclines, is a 9-t-butylglycylamido derivate of minocycline. in this study, the in vitro activity of tigecycline in combination with gentamicin, streptomycin, rifampin, co-trimoxazole, levofloxacin, and minocycline was investigated using the checkerboard method to evaluate 16 Brucella melitensis isolates. The time-kill method was used to determine the bactericidal activities of combinations of tigecycline with rifampin, gentamicin, and levofloxacin, which were found (via the checkerboard method) to have a synergistic effect in combinations with tigecycline. Using the checkerboard method, combinations of rifampin, gentamicin, and levofloxacin with tigecycline showed synergistic effects against 5 (31.2%), 3 (18.9%), and 8 (50%) of the isolates. No synergy was observed with tigecycline in combination with minocycline, streptomycin, or co-trimoxazole. tigecycline with gentamicin achieved the earliest complete killing at 4x miC (in 6 h), while complete killing with the other combinations was delayed up to 24 h. the time-kill method showed that the combination of tigecycline and levofloxacin had an antagonistic effect, while the checkerboard method detected synergy and no interaction effects. these data suggest that a combination regimen of tigecycline with gentamicin and rifampin may be a good choice for treating brucellosis.

Francisella tularensis is the cause of the zoonotic disease tularemia and is classified among highly pathogenic bacteria (HPB) due to its low infection dose and potential for airborne transmission. In the case of HBP, there is a pressing need for rapid, accurate and reliable identification. Phenotypic identification of Francisella species is inappropriate for clinical microbiology laboratories because it is time-consuming, hazardous and subject to variable interpretation. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was recently evaluated as a useful tool for the rapid identification of a variety of microorganisms. In this study, we evaluated the use of MALDI-TOF MS for the rapid identification of Francisella tularensis and differentiation of its subspecies. Using national collection of Francisella isolates from the National Tularemia Reference Laboratory (Public Health Institution of Turkey, Ankara), a total of 75 clinical isolates were investigated by species and subspecies-specific polymerase chain reaction (PCR) test and MALDI-TOF MS. All isolates were originally identified as F. tularensis subsp. holarctica according to region of difference 1 (RD1) subspecies-specific PCR results. For all isolates MALDI-TOF MS provided results in concordance with subspecies-specific PCR analysis. Although PCR-based methods are effective in identifying Francisella species, they are labor-intensive and take longer periods of time to obtain the results when compared with MALDI-TOF MS. MALDI-TOF MS appeared to be a rapid, reliable and cost-effective identification technique for Francisella spp. Shorter analysis time and low cost make this an appealing new option in microbiology laboratories.

Tularemia has become a re-emerging zoonotic disease in Turkey recently. The aims of this study were to determine the seroprevalence of tularemia in humans and their animals living in rural risky areas of our region and to investigate the risk factors. Between January and July 2012, people living in rural areas of Van province (located at eastern part of Turkey) and their domestic animals were included in the study. The sample size was determined by using cluster sampling method like in an event with known prevalence and planned as a cross-sectional epidemiological study. Proportional random sampling method was used to determine which individuals will be included in the study. Presence of tularemia antibodies in the sera of a total 495 voluntary persons (343 female, 152 male; age range: 18-79 years, mean age: 40.61) and their 171 animals (40 cattle, 124 sheep and 7 goats) were screened by microagglutination test using safranin O-stained F.tularensis antigen (Public Health Agency of Turkey). For the evaluation of cross-reactivity between Brucella spp., tularemia positive serum samples were also tested with brucella microagglutination test. Among human and animal samples, 11.9% (59/495) and 44% (76/171) yielded positive results with the titers of ≥ 1:20 in F.tularensis microagglutination test, respectively. However, 69.5% (41/59) of human sera and 78.9% (60/76) of animal sera demonstrated equal or higher titers in the brucella test, so those sera were considered as cross-reactive. After exclusion of these sera, the seroprevalence for F.tularensis were calculated as 3.6% (18/495) for humans and 9.4% (16/171) for animals. Among the 16 animals with positive results, 12 were sheep, three were cattle and one was goat. The difference between seropositivity rates among the domestic animal species was not statistically significant (p> 0.05). In addition, no statistically significant differences were found between risk factors including insect bite, tick bite, contact with rodents, eating the meat of hunted animals (rabbit), having pet (cat) in home (p> 0.05). In this study, the rate of tularemia seropositivity among humans was similar to the results of previous studies which were performed in our country; however the seropositivity rate of tularemia among domestic animals in our study was higher than the results of a few studies which were conducted on domestic animals. In conclusion, preventive procedures and precautions must be taken into consideration to control the transmission of the infection.

Summary Objectives We aimed to determine the seroprevalence of Crimean-Congo hemorrhagic fever (CCHF) virus, Brucella spp , and Coxiella burnetii among veterinarians in a highly endemic and a non-endemic region for these infections in Turkey. Methods The antibody levels against these three infections were surveyed. Eighty-three veterinarians were included from two distinct geographic regions. Results CCHF IgG positivity (3% vs. 0%) and Brucella agglutination titer of ≥1/160 (33% vs. 5%) were more common in the endemic region, whereas the rates of Coxiella burnetii antibodies were similar (7% and 8%). A history of tick bite was significantly more common in the endemic region (35% vs. 12%, p =0.011). A multivariate analysis was performed among the veterinarians living in the endemic area, and percutaneous injuries were found to be associated with Brucella infection (OR 1.8, CI 1.09–3, p =0.022). Conclusions Veterinarians should protect themselves against tick bites, and should use masks to prevent transmission by inhalation of zoonotic infections in endemic countries.

Limited knowledge is available regarding tularemia in pregnancy. A total of seven tularemia cases in pregnant women have been published in the literature. This report presents three new cases. Two of these cases improved without any treatment. The third case was treated with gentamicin. All three pregnancies reached full term without complication for either mother or child.

In many parts of the world, brucellosis has significantly decreased, but it is still a problem in some regions of Turkey. Totally, 11 Brucella strains isolated from the blood cultures of patients with presumptive diagnosis of brucellosis were identified to species and biovar level. For species identification, (i) CO2 requirements of isolates; (ii) detection of urease activation; (iii) detection of H2S production; (iv) dye sensitivities (thionine and basic fuchsin); (v) susceptibility to Brucella phage and (vi) agglutination with monospecific antisera (A and M) were performed. Ten out of 11 isolates were identified as B. melitensis biovar 3 and one out of 11 were identified as B. melitensis biovar 1. The sensitivity of isolates to antibiotics was determined by the E-test. All isolates were found to be sensitive to doxycycline, rifampin, ciprofloxacin and cephtriaxone. Only one out of 11 isolates was found to be semi-sensitive to trimethoprim/sulfamethoxazole (cotrimoxazole). (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Objective. To determine the epidemiological and the clinical characteristics of bacteremic brucellosis. Methods . A prospective study, performed in the First Infectious Diseases Clinic of Ankara Numune Education and Research Hospital. All the patients had positive culture result for Brucella spp. Results . Fifty-four acute bacteremic brucellosis cases were included. The majority of patients (76%) were from rural Anatolia. Brucella melitensis serotypes were more common than Brucella abortus (83% versus 17%). Fever and arthralgia were the most common symptoms. The number of patients with back pain and arthralgia was higher in B. abortus infected group ( p =0.014 and p =0.009). Conclusions . B. melitensis is the most common subtype of Brucella infection in Turkey. The infections with B. abortus spp. are not less severe than the infections with B. melitensis.

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