Application of Hormonal and Single Multiplex PCR Assays for Detection of Freemartinism in a Horned Goat

ABSTRACT The present study describes a rare case of freemartinism of the 60XX/60XY chimera type in an adult 7-year-old horned, mixed Lori breed phenotypically doe. Signs of masculinisation were evident in both the behavioral patterns and body conformation, but there was no structure similar to penis and hypospadias was observed in this goat. External genital organs were similar to that of a female one. In this study, a single multiplex PCR assay was used to determine the freemartinism status (XX-XY blood chimerism) based on semi-quantitatively co amplify a sex-based polymorphism in the amelogenin locus (AMX and AMY). In addition, serum concentrations of anti-Mullerian hormone (AMH), testosterone and progesterone hormones of this horned intersex goat were evaluated. Serum AMH levels in the present case (0.2 ng/ml) were similar to male (0.2 ng/ml) and lower than female (0.6 ng/ml) goat. The serum concentrations of testosterone (0.2 ng/ml) were remarkably high in comparison to normal control male and female ones (0.02 ng/ml). Progesterone level in the intersex goat (0.4 ng/ml) was close to control buck (0.7 ng/ml) and remarkably lower than control doe (10 ng/ml). The results of the present study showed that the use of hormonal pattern, especially AMH level, and single multiplex PCR technique provide an easy and alternative approaches for indication of XX/XY chimerism or mosaicism in intersex freemartin goats. Key Words: Freemartin, XX/XY chimerism, intersex, horned goat, single multiplex PCR, anti-mullerian hormone OZET BOYNUZLU BIR KECIDE FREEMARTINIZMIN HORMONAL VE TEKLI MULTIPLEKS PCR ANALIZ UYGULAMALARI ILE TESPITI Bu calismada, nadir bir olgu olan freemartinizmin disi fenotipine sahip Lori irki melezi 7 yasindaki boynuzlu ergin bir kecide 60XX/XY kimera tipi bildirilmektedir. Davranis bicimlerinde ve vucut yapisinda maskulinizasyon belirtileri bulunur iken, penis veya hypospadias gozlenmemistir. Dis genital organlar disi keci ile benzerlik gostermektedir. Bu calismada amelogonin lokusundaki (AMX ve AMY) cinsiyete bagli polimorfizmin semi-kantitatif olarak cogaltilmasi ile freemartinizm durumunun (XX-XY kan kimerizmi) belirlenmesinde tekli multipleks PCR yontemi kullanilmistir. Ayrica anti-Mullerian hormon (AMH), testesteron ve progesteron hormonlarinin serum konsantrasyonlari da degerlendirmeye alinmistir. Mevcut vakadaki serum AMH duzeyleri (0,2 ng/ml) tekeye benzer (0,2 ng/ml) olarak, disi keciden (0,6 ng/ml) ise dusuk olarak saptanmistir. Serum testesteron konsantrasyonu (0,2 ng/ml) kontrol tekeye ve disi keciye (0,02 ng/ml) gore onemli duzeyde yuksek bulunmustur. Interseks kecide progesteron duzeyi (0,4 ng/ml) kontrol tekedeki duzeye (0,7 ng/ml) yakin, kontrol disi kecideki duzeyden (10 ng/ml) ise belirgin olarak dusuktur. Mevcut calismanin sonuclari interseks freemartin kecilerde XX/XY kimerizmi veya mozaizmini belirlemede AMY duzeyi ve tekli multipleks PCR tekniginin kolay ve alternatif bir yontem oldugunu ortaya koymaktadir. Anahtar Kelimeler: Freemartin, XX/XY kimerizmi, interseks, boynuzlu keci, tekli mutlipleks PCR, anti-mullerian hormon