BİLAL AKYÜZ, DAVUT BAYRAM, Kutlay GÜRBULAK

Polimeraz zincir reaksiyonu (PZR) ile sığırlardaki freemartinismus sendromunun tanısı

Freemartinismus sendromu, farklı cinsiyette yavruların bulunduğu ikiz gebeliklerde erkek yavruya ait kan kök hücrelerinin ve anti-Müllerian faktörü hormonunun dişi ikizin dolaşımına karışması sonucu dişi yavruların steril olması durumudur. Erkek ikizi olan dişi buzağılarda %82.5 ve %92 oranında freemartinusmus gelişebilir. Freemartin bireylerin belirlenmesinde çeşitli yöntemler kullanılmaktadır. Ancak freemartin yavruların erken dönemde belirlenmesi, yavrunun damızlık olarak kullanılıp kullanılmaması için önemlidir. Erken dönemde yavrunun freemartin olup, olmadığının belirlenmesi sitogenetik veya Y spesifik primerlerin kullanıldığı moleküler yöntemlerle yapılabilir. Ancak hassasiyeti, uygulama kolaylığı ve tanı süresinin kısalığı yönünden moleküler yöntemler tercih edilir. Bu çalışmada, erkek ikizi olan üç dişi buzağının freemartinismus durumları polimeraz zincir reaksiyonu (PZR) ile araştırılmıştır. PZR işlemi Y kromozomuna özel iki primer seti kullanılarak yapılmıştır. Ayrıca bu yavruların vajina uzunlukları ve internal üreme kanalı transrektal ultrason ile incelenmiştir. PZR işlemi sonunda erkek ikiz kardeşi olan dişi yavruların ikisinin Y kromozomuna sahip olduğu, bir buzağının ise Y kromozomu taşımadığı belirlenmiştir. Aynı zamanda bu yavrunun vajina uzunluğunun incelenen diğer buzağılardan daha uzun olduğu (9 cm) ve ultrasonografik incelemeler sonunda serviksin bulunduğu görülmüştür. Bu çalışmadan elde edilen bulgulara göre erkek ikizi olan dişi yavruların ve tekrarlayan tohumlamalarda gebe kalmayan damızlık dişi sığırların freemartinismus yönünden araştırılmasında PZR yönteminin kullanılabileceği sonucuna varılmıştır.

Anahtar Kelimeler:

teşhis, y kromozom, buzağı, sığır, tanı teknikleri, freemartinizm, polimeraz zincir reaksiyonu

Diagnosis of freemartinism syndrome by polymerase chain reaction (PCR) in cattle

Freemartinism syndrome is a case of sterility in female twin of spring when twin pregnancy exists with the dif erent sexes, and caused by involvement of male twin’s blood stem cell factor and anti-Mullerian hormone into female twin. Freemartinism may develop in female twin at a rate of 82.5% and 92% in twins with dif erent sexes. Dif erent methods are used to determine freemartinism syndrome. However, early identification of freemartinism is very important to determine the potential of calf for breeding purposes. Early identification of freemartinism could be accomplished by cytogenetic and molecular methods via Y-specific primers. However, molecular methods are preferred due to their accuracy, ease of application and shorter duration of process. In this study, freemartinism status of three female calves born with twin male calves was investigated by polymerase chain reaction (PCR). PCR process was carried out by using two primer sets, specific to Y-chromosome. Moreover, the length of the vagina and reproductive tract was examined by transrectal ultrasonography. PCR process revealed that the two female calves had the Y chromosome, whereas the other calf did not have the Y chromosome. Furthermore, length of vagina is longer (9 cm) in a calf not having Y chromosome than the other calves, and ultrasound examination indicated the presence of cervix in a calf without Y chromosome. According to current results, it was concluded that PCR technique could be used to investigate freemartinism cases in female twin calves with male twins and non-pregnant females following recurrent breeding.

Keywords:

diagnosis, Y chromosome, calves, cattle, diagnostic techniques, freemartinism, polymerase chain reaction,

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