Trakya bölgesinde tarla farelerinde kültür, seroloji ve moleküler yöntemlerle Francisella tularensisin aranması
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Dosyalar
Tarih
2013
Yazarlar
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Yayıncı
Trakya Üniversitesi Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Tularemi Türkiye'de 1936 yılından beri bildirilmektedir ve etkenin bulaşmasında farelerin rolü üzerinde durulmaktadır. Bugüne kadar etkenin bulaşmasında farelerin rolü kesin olarak gösterilememiştir. Çalışmanın amacı kültür, seroloji ve moleküler metodlar kullanarak tarla farelerinde Francisella tularensis'in varlığını araştırmaktır. Trakya Bölgesinden dört köy seçilerek toplam 126 adet fare kapanı depolara, ahırlara, kuyu kenarları, su depoları ve dere kenarlarına yakın bölgelere kuruldu. Canlı olan farelere karbondioksit ile anestezi uygulandı ve kalpten doğrudan kanları alındı. Daha sonra tüm farelerin karaciğer ve dalak dokuları sınıf-2 güvenlik kabini içinde aseptik şartlarda çıkarıldı. Bu dokular Francis besiyerlerine ekilerek hem normal atmosferde hem de %5 karbondioksit içeren etüvde 7 gün boyunca bekletildi. Farelerin serumlarına tularemi mikroaglütinasyon testi uygulandı. Son olarak farelerin hem karaciğer hem dalak dokularına eş zamanlı polimeraz zincir reaksiyonu testi (RT-PCR) yapıldı. Toplam 19 adet tarla faresi yakalandı. Bunların on tanesi Apodemus flavicollis, yedi tanesi Mus macedonicus, iki tanesi ise Mus musculus türlerine aitti. Bu 19 farenin 11'i canlı diğerleri ölüydü. Farelerin karaciğer ve dalak kültürlerinde F. tularensis izole edilemedi. Tüm farelerin serolojik testleri negatifti. 1936 yılında Türkiye?de ilk tularemi olgularının görüldüğü Kaynarca?dan yakalanan iki farenin dalak dokusunda RT-PCR testinde pozitiflik tespit edildi. Bunlardan biri dişi bir Mus macedonicus, diğeri ise erkek bir Apodemus flavicollis'ti. Bu çalışma ile doğal ortamdaki farelerin tularemi etkenini taşıdıkları Türkiye'de ilk kez kesin olarak gösterilmiştir. Etken 1936 yılından bu yana Kaynarca'da varlığını sürdürmekte olabilir ve tularemi için potansiyel tehlike oluşturmaktadır.
Abstract
Tularemia has been reporting in Turkey since 1936 and focusing on the role of mice in the transmission of the causative agent. Unfortunately, the role of mice in the transmission of the disease to date could not be exactly confirmed. The aim of this study is to investigate the presence of Francisella tularensis by using culture, serology and molecular methods in mice. Four villages in Thrace Region were visited and totally 126 mice traps were established in warehouses, barns, close areas near wells, water tanks and creeks. The live mice were anesthetized by carbondioxide and their bloods were taken directly from the heart. Then liver and spleen tissues were taken from all mice by aseptic methods in class-2 safety cabinet. These tissues were cultured in Francis media for 7 days in both of normal atmosphere and 5% carbondioxide incubator in 37°C. Tularemia microaglutination test was applied to mice sera. Finally, real time polymerase chain reaction test (RT-PCR) was performed to both of liver and spleen tissues of the mice. Totally 19 mice were captured. Ten mice were Apodemus flavicollis, seven were Mus macedonicus, two were Mus musculus. Of 19 mice, 11 were alive but the others were dead. There was no Francisella tularensis isolation in mice liver and spleen tissue cultures. Serological tests were negative in all mice. In RT-PCR, positivity were detected in spleen tissues of two mice which were captured from Kaynarca where first reported tularemia cases in Turkey in 1936. One of them was a female Mus macedonicus, the other was a male Apodemus flavicollis. In this study, it was firstly shown that mice in natural environment have causative agent of tularemia in Turkey. The agent may persist since 1936 in Kaynarca and have a potential danger for tularemia.
Abstract
Tularemia has been reporting in Turkey since 1936 and focusing on the role of mice in the transmission of the causative agent. Unfortunately, the role of mice in the transmission of the disease to date could not be exactly confirmed. The aim of this study is to investigate the presence of Francisella tularensis by using culture, serology and molecular methods in mice. Four villages in Thrace Region were visited and totally 126 mice traps were established in warehouses, barns, close areas near wells, water tanks and creeks. The live mice were anesthetized by carbondioxide and their bloods were taken directly from the heart. Then liver and spleen tissues were taken from all mice by aseptic methods in class-2 safety cabinet. These tissues were cultured in Francis media for 7 days in both of normal atmosphere and 5% carbondioxide incubator in 37°C. Tularemia microaglutination test was applied to mice sera. Finally, real time polymerase chain reaction test (RT-PCR) was performed to both of liver and spleen tissues of the mice. Totally 19 mice were captured. Ten mice were Apodemus flavicollis, seven were Mus macedonicus, two were Mus musculus. Of 19 mice, 11 were alive but the others were dead. There was no Francisella tularensis isolation in mice liver and spleen tissue cultures. Serological tests were negative in all mice. In RT-PCR, positivity were detected in spleen tissues of two mice which were captured from Kaynarca where first reported tularemia cases in Turkey in 1936. One of them was a female Mus macedonicus, the other was a male Apodemus flavicollis. In this study, it was firstly shown that mice in natural environment have causative agent of tularemia in Turkey. The agent may persist since 1936 in Kaynarca and have a potential danger for tularemia.
Açıklama
Tıpta Uzmanlık Tezi
Anahtar Kelimeler
Tularemi, Seroloji ve Moleküler Yöntemler, Kültür, Francisella Tularensis, Zoonoz
