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Öğe Aggregatibacter actinomycetemcomitans Kaynaklı Olası Enfektif Endokardit Olgusuna Mikrobiyolojik Yaklaşım(2016) Kuloğlu, Hüsnüye Figen; Karadenizli, Aynur; Ünlü, Selahattin; Kuşkucu, Mert Ahmet; Gürcan, ŞabanAggregatibacter (Actinobacillus) actinomycetemcomitans, küçük, gram-negatif boyanan kokobasil morfolojisinde, zor üreyen, müşkülpesent bir bakteri olup, sıklıkla ağız boşluğunda kolonize olmaktadır. İzolasyonunun güç olmasının da etkisiyle, çok sık karşılaşılmayan fakat diş ile ilgili enfeksiyonlar ve özellikle protez kapak öyküsü olan kişilerde enfektif endokardit (EE) etkeni olabilen bir bakteridir. Bu raporda, aort kapak replasmanı öyküsü olan bir hastada gelişen A.actinomycetemcomitans kaynaklı olası EE olgusu sunulmaktadır. Otuz altı yaşındaki erkek hasta, üşüme, titreme, aralıklı yüksek ateş, halsizlik, yaygın eklem ağrısı ve kilo kaybı (yaklaşık 20 kg) şikâyetleriyle Trakya Üniversitesi Sağlık Araştırma ve Uygulama Merkezine başvurmuştur. Hastanın izlemi sürecinde, üç hafta arayla alınan kan kültürlerinde aynı gram-negatif kokobasil morfolojisinde bakteri üremesi saptanmıştır. Olguya, bir majör (iki farklı kan kültüründe EE etkeni olabilecek tipik mikroorganizmaların üremesi) ve iki minör (protez kapak varlığı ve ateş yüksekliği) kriteri karşılaması nedeniyle olası EE tanısı konulmuştur. İzole edilen suş konvansiyonel yöntemlerle tanımlanamamış, VITEK 2 (bioMerieux, Fransa) cihazı ile yapılan tanımlamada Francisella tularensis olarak isimlendirilmiş, fakat gerçek zamanlı Taqman polimeraz zincir reaksiyonu ile bu sonuç doğrulanamadığından, bakteri MALDITOF kütle spektrofotometre yöntemi ile tanımlanmaya çalışılmıştır. Bu yöntem ile yapılan ilk çalışmada, bakteri Shigella dysenteriae olarak, ertesi gün yapılan test tekrarı sonucunda ise A.actinomycetemcomitans olarak isimlendirilmiştir. Birbirinden farklı çıkan sonuçları netleştirmek için 16S ve 23S ribozomal DNA dizi analizi uygulanmış ve sonuçta izolat A.actinomycetemcomitans olarak tanımlanmıştır. Olgunun ilk şüpheli etken isimlendirme sonucu (F.tularensis) dikkate alınarak başlanan doksisiklin (2 x 100 mg po, 20 gün) ve streptomisin (2 x 10 mg/kg im, 10 gün) tedavisinin beşinci gününde kan kültüründe üreme olmamış ve hastanın yakınmaları gerilemiştir. Tedavinin 21. gününden itibaren doksisikline rifampisin eklenerek tedaviye devam edilmiş ve hasta şifa ile taburcu edilmiştir. Sonuç olarak, A.actinomycetemcomitans ve bunun gibi nadir görülen hastalık etkenlerinin isimlendirmeleri, laboratuvarların imkânları dâhilinde zor ve zaman alıcı olabilir. Nadir izolatların isimlendirmesinde karşılaşılan sorunları aşmak için, özellikle de olgunun kliniği ile uyumun olmadığı durumlarda, klasik yöntemlerin yanında diğer tanı yöntemlerinin de kullanılması zorunlu hale gelebilir. Bu olgu, EE hastalarında A.actinomycetemcomitans'ın nadiren izole edilen bir etken olması ve otomatize sistemler ile mikroorganizmaların isimlendirmesinde karşılaşılabilecek çeşitli sorunlara örnek olabilecek bir olgu olması nedeniyle sunulmuşturÖğe Characteristics of the Turkish isolates of Francisella tularensis(Natl Inst Infectious Diseases, 2008) Gurcan, Saban; Karabay, Oguz; Karadenizli, Aynur; Karagol, Cigdem; Kantardjiev, Todor; Ivanov, Ivan N.In this study, cultures of patients with tularemia were evaluated, and antimicrobial susceptibilities of two Francisella tularensis strains were tested by disk diffusion and E-test methods. A high-resolution multiple-locus variable-number tandem repeat analysis (MLVA) comprising six variable-number tandem repeat loci was applied to elucidate the genetic relatedness among Turkish and Bulgarian isolates which were isolated in a recent outbreak. The patients were diagnosed in two outbreaks in two cities of Turkey in 2005 and 2006. A total of 16 samples from 12 patients were cultured, and PCR tests were carried out on 15 samples that were positive in five lymph node aspirates and two soft tissue aspirates. F. tularensis was isolated from the lymph nodes of two patients. Aminoglycosides, quinolones, chloramphenicole, tetracyclines, nitrofurantoin, and rifampicin inhibited growth of the isolates. The Turkish isolates appeared to share a common MLVA pattern with one of the four Bulgarian outbreak genotypes.Öğe Investigation of the Presence of Francisella tularensis by Culture, Serology and Molecular Methods in Mice of Thrace Region, Turkey(Ankara Microbiology Soc, 2014) Unal Yilmaz, Gulizar; Gurcan, Saban; Ozkan, Beytullah; Karadenizli, AynurTularemia is a disease that has been reported in Turkey since 1936. Although mice are considered to have a role in the transmission of Francisella tularensis to man, this has not been exactly confirmed yet. The aim of this study was to investigate the presence of F. tularensis in mice by using culture, serology and molecular methods. For this purpose, four villages (Edirne-Demirkoy, Kirklareli-Kaynarca, Tekirdag-Muzruplu, Tekirdag-Sinanli) were selected in Thrace Region of Turkey where tularemia cases had been reported previously. A total of 126 live-catch mouse traps were established in warehouses, barns, areas near wells, water tanks and creeks in the villages in December 2012. Traps were kept overnight and the next day the animals collected were identified at species-level. The live-captured mice were anesthetized and their heart blood samples were obtained. Subsequently, liver and spleen tissues were removed from every mouse under aseptic conditions in the class-2 safety cabinet. These tissues were cultivated in Francis medium containing 5% sheep blood, 0.1% cystein, 1% glucose and incubated for seven days in both normal atmosphere and 5% carbondioxide incubator at 37 degrees C. Tularemia microagglutination test was performed by using the sera which were obtained from live-captured mice. Finally, DNAs were isolated from both liver and spleen tissues of mice, and real-time polymerase chain reaction (Tularemia RT-PCR; Public Health Agency of Turkey, Ankara) were performed. In our study, a total of 19 mice were captured and of these 11 were alive. Ten mice were identified as Apodemus flavicollis, seven were Mus macedonicus and two were Mus musculus. There were no Francisella tularensis isolation in the cultures of mice liver and spleen tissues. Serological tests yielded negative results for 10 mice whose serum samples could be obtained. In RT-PCR, positivity were detected in spleen tissues of two mice which were captured from Kaynarca where first tularemia cases in Turkey in 1936 were reported but has no report from then on. One of them was a live female Mus macedonicus, and the other was a dead male Apodemus flavicollis. In quantitative evaluation, number of microorganism per organ were calculated as 4 x 10(3) cfu/spleen in Mus macedonicus and 4 x 10(4) cfu/spleen in Apodemus flavicollis. This is the first study in Turkey indicating that the mice in natural environment harbored Etularensis. In conclusion, the results of this study indicated that the agent of tularemia has been retained since 1936 in Kaynarca region and this persistence might present a potential risk for tularemia epidemics.Öğe Investigation of Tularemia Incidence and Presence of Francisella tularensis in Streams/Mains Water in a Risky Region of Thrace(Aves, 2019) Ugur, Mediha; Gurcan, Saban; Eskiocak, Muzaffer; Karadenizli, AynurObjective: Tularemia was first detected in Thrace region in our country and the outbreaks continued in the region over the following years. The fact that the agent has been identified in mice around Kaynarca in 2012 suggests the disease poses a risk for our region. Aim of this study was to investigate tularemia incidence and presence of Francisella tularensis in streams/mains water in a risky region of Thrace. Methods: In this study, seropositivity for tularemia was investigated in 13 villages, and 1 town in risky areas of the Thrace region. In January 2016, blood was drawn from 746 people and tularemia microagglutination tests were applied. Seropositivity was not detected. In December, 464 of 746 people were reached. Seroconversion was not observed. In addition, culture and polymerase chain reaction (PCR) procedures were applied to specimens collected from mains water and streams in 13 villages and 1 town. Results: The causative agent wasn't isolated from the cultures but F. tularensis DNAs were detected by PCR method in 2 stream, and 3 mains water samples. One of the streams passed through the village of Celaliye, which was very close to Kaynarca, where tularemia cases were seen in the past. The other was farther, passing through the Kavakli town in which no cases has been reported. The mains water which were positive were from Hamzabey, Ceylankoy, and Tatarkoy villages located around Kaynarca. Molecular examination after chlorination was repeated in the water sources in which positivity was detected, and it was seen that the agent was eliminated. Conclusions: In this study, incidence was calculated as zero, although the causative agent was found in the water. Although no seropositivity was detected, the detection of the agent by PCR in 5 water samples showed that the agents in the nature could reach the water resources. It has been observed that surveillance studies in risky areas could be effective in preventing possible outbreaks.Öğe Microbiological Approach to a Possible Infective Endocarditis Case Caused by Aggregatibacter actinomycetemcomitans(Ankara Microbiology Soc, 2016) Gurcan, Saban; Unlu, Salahattin; Kuloglu, Figen; Karadenizli, Aynur; Kuskucu, Mert AhmetAggregatibacter (Actinobacillus) actinomycetemcomitans, a small, gram-negative coccobacillus that grows slow and fastidious, is generally colonized in the oral cavity. It is a rarely seen bacterium because of the difficulty of isolation but it can be a causative agent for dental infections and infective endocarditis (IE) particularly in the persons having prosthetic heart valves. In this report, a possible IE case caused by A.actinomycetemcomitans in a patient with aortic valve replacement has been presented. A 36-year-old man has admitted to Trakya University Hospital, Health Center for Medical Research and Practice, with the complaints of chills, malaise, intermittent fever, severe arthralgia and weight loss (20 kg). During his follow-up period, the blood cultures that were obtained three week intervals yielded the identical gram negative coccobacilli morphology. The patient was then diagnosed as possible IE on the basis of having one major (growth of the typical microorganisms that may cause IE in two different blood cultures) and two minor (presence of prosthetic valve and high fever) criterias. The isolate could not be identified with conventional methods, while it was identified as Francisella tularensis with VITEK 2 (bioMerieux, France) system. Hence this identification was not confirmed by real-time Taqman polymerase chain reaction, so MALDI-TOF mass spectrometry was used to identify this bacteria. In the first run of the study, the isolate was named as Shigella dysenteriae initially, however when it was retested the next day it was identified as A.actinomycetemcomitans. In order to enlighten these conflicting results, 16S and 23S ribosomal DNA sequence analysis was performed, and consequently the bacterium was identified as A.actinomycetemcomitans. Doxycycline (2 x 100 mg po, 20 days) and streptomycin (2 x 10 mg/kg im, 10 days) therapy were initiated, considering the initial suspicious identification (F.tularensis), and on the fifth day of therapy the blood culture was negative with the regression of patient's complaints. Therapy continued with the addition of rifampicin to doxycycline from the 21(st) day and the patient discharged with cure. As a result, the identification of an exceptional bacterium like A.actinomycetemcomitans may be difficult and time-consuming in certain laboratory facilities. So, the use of different identification methods in addition to classical methods are needed to overcome such a problem, especially for uncommon isolates and clinically discordant cases. This case was presented because A.actinomycetemcomitans is a rare etiological agent for IE patients and it could be a good example to draw attention to the problem when identifying the organism using automatized identification systems.Öğe Trakya Bölgesi’nde farelerde kültür, seroloji ve moleküler yöntemlerle francisella tularensis varlığının aranması(2014) Gürcan, Şaban; Karadenizli, Aynur; Ünal, Gülizar Yılmaz; Özkan, BeytullahTularemi Türkiye’de 1936 yılından beri bildirilen bir hastalıktır ve etkenin insanlara bulaşmasında farelerin rolü üzerinde durulmaktadır. Ancak bugüne kadar etkenin bulaşmasında farelerin rolü kesin olarak gösterilememiştir. Bu çalışmanın amacı, farelerde Francisella tularensis varlığının kültür, seroloji ve moleküler yöntemlerle araştırılmasıdır. Çalışma için Trakya Bölgesi’nden, daha önce tularemi olgularının bildirildiği dört köy (Edirne-Demirköy, Kırklareli-Kaynarca, Tekirdağ-Muzruplu, Tekirdağ-Sinanlı) seçilmiş ve Aralık 2012 tarihinde bu köylere gidilerek uygun depolar, ahırlar, ambarlar, dere ve kuyu kenarları, yemlikler ve su depoları gibi bölgelere toplam 126 adet canlı fare yakalama kapanları kurulmuştur. Kapanlar bir gece bekletildikten sonra toplanmış ve yakalanan farelerin türleri belirlenmiştir. Canlı olarak yakalanan farelerden anestezi altında kalp kanları alınmış; daha sonra tüm farelerin karaciğer ve dalak dokuları sınıf-2 güvenlik kabini içinde aseptik şartlarda çıkartılmıştır. Bu dokular %5 koyun kanı, %0.1 sistein ve %1 glukoz içeren Francis besiyerlerine ekilmiş; besiyerleri hem normal atmosferde hem de %5 karbondioksit içeren etüvde yedi gün boyunca inkübe edilmiştir. Canlı olarak yakalanıp kanları alınabilen farelerin serumlarında F.tularensis’e özgül antikorlar mikroaglütinasyon testi ile araştırılmıştır. Çalışmada ayrıca, farelerin karaciğer ve dalak dokularından DNA izolasyonu yapılmış ve F.tularensis ISFtu2 genine özgül primerler ve prob kullanılarak gerçek zamanlı polimeraz zincir reaksiyonu (Tularemi RT-PCR; Türkiye Halk Sağlığı Kurumu, Ankara) uygulanmıştır. Çalışmamızda 11’i canlı olmak üzere toplam 19 adet tarla faresi yakalanmış; bunların 10’unun Apodemus flavicollis, yedisinin Mus macedonicus, ikisinin ise Mus musculus türlerine ait olduğu belirlenmiştir. Farelerin karaciğer ve dalak dokularından yapılan kültürlerin hiçbirisinden F.tularensis izole edilememiş; serolojik yöntemin uygulanabildiği 10 fare serumu ise negatif sonuç vermiştir. Buna karşın RT-PCR testi ile, 1936 yılında Türkiye’de ilk tularemi olgularının görüldüğü ve daha sonra başka olgunun bildirilmediği Kaynarca beldesinde yakalanan iki farenin dalak dokusunda pozitiflik tespit edilmiştir. Bunlardan biri canlı olarak yakalanan dişi bir Mus macedonicus, diğeri ise ölü olarak yakalanan erkek bir Apodemus flavicollis’tir. Yapılan kantitatif değerlendirmede, organ başına bakteri sayısı Mus macedonicus için 4 x 103 cfu/dalak, Apodemus flavicollis için 4 x 104 cfu/dalak olarak hesaplanmıştır. Sonuç olarak, bu çalışma ile doğal ortamdaki farelerin tularemi etkenini taşıdıkları Türkiye’de ilk kez gösterilmiş ve 1936 yılından bu yana F.tularensis’in Kaynarca’da varlığını sürdürerek, salgınlar için potansiyel risk oluşturmaya devam ettiği düşünülmüştür.Öğe Tularemia as a result of outdoor activities for children in the countryside(Tubitak Scientific & Technological Research Council Turkey, 2012) Gurcan, Saban; Saracoglu, Gamze Varol; Karadenizli, Aynur; Ozkayin, Emine Nese; Ozturk, Semsi Zafer; Cicek, Cemal; Vatansever, BinayAim: To investigate the features of a new tularemia outbreak that occurred in the Thrace region. Materials and methods: The research team visited the village after the identification of the index case. Serum and throat samples were taken from 41 villagers who were examined, and environmental samples were taken in order to identify the source of the outbreak. Culture, serology, and molecular methods were used to search for Francisella tularensis in these samples. Results: A total of 8 children were diagnosed with tularemia. The adults and all of the other children were seronegative for tularemia. All of the patients had a history of swimming in a pool filled with water from a local stream, and contact with stream water was calculated to increase the risk of developing the disease 9.3-fold. Polymerase chain reaction analysis was positive in a lymph node aspirate of the index case and in the home tap water of 3 patients as well as in the spring water and stream water in the village. Francisella tularensis could not be isolated from any culture of samples. Interestingly, the waterborne tularemia outbreak affected only children. Conclusion: Although tularemia has been not reported from Tekirdag Province for 74 years, the disease reemerged in the region due to the removal of hygienic measures. These clues may signify that the agent had maintained its presence in the region for many years.Öğe Tularemia re-emerging in European part of Turkey after 60 years(Natl Inst Infectious Diseases, 2006) Gurcan, Saban; Eskiocak, Muzaffer; Varol, Gamze; Uzun, Cem; Tatman-Otkun, Muserref; Sakru, Nermin; Karadenizli, AynurThe aim of this study was to investigate a tularemia outbreak in the Thrace region of Turkey. The outbreak occurred in Demirkoy village of Edirne, in 2005. Of 400 villagers, 266 were examined and their sera were taken. Throat swabs and lymph node aspirates were cultured. Specific antibodies in patients and domestic animals were screened by a microagglutination test. PCR assays and cultures of the samples of patients, animal tissues, and water sources were performed,. along with active surveillance to identify risk factors. Seven out of 10 cases were diagnosed as oropharyngeal form; the remaining three patients were asymptomatic. The cultures for tularemia were negative; however, PCR assays were positive in one lymph node aspirate and in water from one spring. Some animals had the specific antibody at low levels. Increased rodent population in the vicinity, exposure to wild rabbits, and drinking from one of the springs were identified as risk factors with the risk ratios (and 95% confidence interval) of 10.5 (10.3-10.7), 6.5 (5.43-7.57), and 2.1 (1.1-2.5), respectively. Therapeutic and preventive measures were taken. When tularemia cases have been detected in a region even a few decades earlier, tularemia should be considered in the differential diagnosis of patients.
