Mehmet NİZAMLIOĞLU, Ercan KURAR, Yusuf ÖZŞENSOY, Müge Doğan BÜLBÜL

DNA teknolojisi ile rendering ürünlerinde tür tespiti: Bir vaka raporu

Deli Dana Hastalığı (BSE) sonrası özellikle ruminant kökenli rendering ürünlerinin hayvan rasyonlarında kullanılması kontrol altına alınmıştır. Gümrük Beyan Belgesinde “hidrolize tüy unu” olarak bildirilen yem materyalinden alınan örnek, T.C. Gümrük Müsteşarlığı Mersin Gümrük Müdürlüğü tarafından gönderilerek hayvansal kökeninin belirlenmesi istenmiştir. İlgili örneğin mikroskobik incelenmesinde kanat ve tüy parçalarının varlığı saptanmıştır. Alternatif olarak, örneklerden standart fenol/kloroform ve dodecyltrimethylammonium bromide (DTAB) yöntemleri ile DNA izolasyonu yapılmıştır. Elde edilen DNA örnekleri kanatlı, ruminant, domuz, at ve karnivor spesifik primerler kullanılarak Polimeraz Zincir Reaksiyonu (PZR) ile yükseltgenmiştir. PZR reaksiyonlarında tavuk, keçi, at, kedi DNA’ları pozitif kontrol, her bir lokus sisteminde DNA negatif PZR reaksiyonları negatif kontrol olarak kullanılmıştır. Tüm PZR ürünleri %2 agaroz jel elektroforezinde ayrıştırılmış ve görüntülenmiştir. İki farklı kanatlı spesifik markör sistemi kullanılarak yapılan PZR analizleri sonucunda, yem örneğinde kanatlı DNA’sının varlığı tespit edilmiştir. Ancak, ruminant, domuz, at ve karnivor DNA varlığı tespit edilememiştir. Bu çalışma, moleküler biyoloji yöntemlerinin yüksek ısılarda muamele edilerek hazırlanan rendering ürünlerinin kökeninin saptanmasında başarıyla kullanılabileceğini göstermektedir.

Identification of animal species in rendering products by DNA technology: A case report.

After bovine spongiform encephalopathy (BSE) outbreak, the use of especially ruminant byproducts in animal rations is under strict control. An animal feedstuff sample was sent from the Turkish Republic, Mersin Customs Directorate in order to identify the animal source. The feedstuff was declared as “hydrolyzed feather meal” in the custom declaration and health certificates. In microscopic inspections, pieces of feathers and hollow shafts were observed. As an alternative method, DNA was extracted by using standard phenol/chloroform and dodecyltrimethylammonium bromide (DTAB) methods. The DNA samples were amplified by Polymerase Chain Reactions (PCR) using poultry, ruminant, swine, horse and carnivore specific oligos. In PCR reactions, chicken, goat, horse and cat DNAs were used as positive controls. DNA free PCR reactions were also amplified as negative control in each system. The resulting PCR products were separated and visualized by electrophoresis on a 2% agarose gel. The findings suggested presence of poultry DNA in the feedstuff sample since two different poultry specific PCR primer pairs resulted in positive PCR products. However, no PCR products were observed in ruminant, swine, horse and carnivore PCR reactions. The results of this study thereby indicated that molecular biology techniques could successfully be used to identify source of heat proceeded animal byproducts.

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