BibTex RIS Kaynak Göster

Abort Olgularının Görüldüğü Süt Sığırı İşletmelerinde Bovine Viral Diyare Virusun Araştırılması

Yıl 2013, Cilt: 10 Sayı: 2, - , 01.06.2013

Öz

The objective of this study is to investigate whether sex distribution and the variance of this distribution areaffected by sex. Sex detection of in vivo produced bovine embryos at different stages was made using nonelectrophoretic PCR method. For sexing; in vivo derived day-seven 65 embryos were used. Embryos were obtainedfrom routine superovulation procedure. After seven days of artificial insemination; embryos were collected bynonsurgical uterine flushing technique. The sex of the embryos was determined; following evaluation of the quality ofembryos. 10-30% of the trophectoderm of the embryo was excised with a micro blade attached to the micromanipulator. Embryo biopsy procedure was carried out in D-PBS solution containing 20% FCS. The biopsy materialswere placed in PCR instrument for lysis and denaturation. After lysis period, DNA amplification process was carried outin 35 cycles. The tubes taken out of the PCR instrument were inspected under UV illumination of 302-312 nm wavelength. Pink fluorescence indicates the presence of a male sample. As a result of the sex determination out of 30embryos; 14 male (46.7%; 14/30), 16 female (53.3%; 16/30) were determined at compact morula stage, out of 35embryos; 21 male (60.0%; 21/35), 14 female (40.0%; 14/35) were determined at blastocyst stage. Even though the rateof males (60%) at blastocyst stage was higher than that at morula stage (46.7%), the difference was not foundstatistically significant (P>0.05).

Kaynakça

  • Avery B, Claus BJ, Madison V, Greve T. Morphological development and sex of bovine in vitro fertilized embryos. Mol Reprod Dev 1992; 32: 265-70.
  • Bredbacka P, Veimala R, Peippo J. Survival of biopsied and sexed bovine demi-embryos. Theriogenology 1994; 41: 1023-31.
  • Bredbacka P, Kankaanpaa A, Peippo, J. PCR sexing of bovine embryos: A simplified protocol. Theriogenology 1995; 44: 167-76. Garcia JF, Nogueira MFG, Pupim F, Pantano T, Visintin JA. Pregnancy rates of blastomere biopsied bovine embryos frozen in ethylene glycol. Theriogenology 1997; 47: 268.
  • Hasler JF, Cardey E, Stokes JE, Bredbacka P. Nonelectrophoretic PCR-sexing of bovine embryos in a commercial environment. Theriogenology 2002; 58: 1457-69. Hassun PA, Mello MRB, Porto LPC, Garcia JF. Bovine embryo sexing by primer extension preamplification polymerase chain reaction. Theriogenology 1999; 51: 398. Jafar SI, Flint APF. Sex selection in mammals: A review. Theriogenology 1996; 46: 191-200. King WA, Yadav BR, Xu KP, Picard L, Sirard MA, Supplizi AV, Betteridge KJ. The sex ratios of bovine embryos produced in vivo and in vitro. Theriogenology 1991; 36: 779-88. Kitiyanant Y, Saikhun J, Siriaroonrat B, Pavasuthipaisit K. Sex determination by polymerase chain reaction and karyotyping of bovine embryos at first cleavage in vitro. Scienceasia 2000; 26: 9-13.
  • Kobayashi J, Sekimoto A, Uchida H, Wada T, Sasaki K, Sasada H, Umezu M, Sato E. Rapid detection of male-spesific DNA sequence in bovine embryos using fluorescence in situ hybridization. Mol Reprod Dev 1998; 51: 390
  • Machaty Z, Paldi A, Csaki T, Varga Z, Kiss I, Barandi Z, Vajta G. Biopsy and sex determination by PCR of IVF bovine embryos. J Reprod Fertil 1993; 98: 467-70.
  • Park JH, Lee JH, Choi KM, Joung SY, Kim JY, Chung GM, Jin DI, Im KS. Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenology 2001; 55: 1843-53.
  • Rens W, Welch GR, Johnson LA. Improved flow cytometric sorting of X and Y chromosome bearing sperm: Substantiol increase in yield of sexed semen. Mol Reprod Dev 1999; 52: 50-6.
  • Shea BF. Determining the sex of bovine embryos using polymerase chain reaction results: A six-year retrospective study. Theriogenology 1999; 51: 841-54.
  • Şendağ S, Aydın İ, Çelik HA. İneklerde prenatal embriyonik/fötal cinsiyetin belirlenmesi. Erciyes Üniv Vet Fak Derg 2005; 2(1): 39Thibier M, Nibart M. The sexing of bovine embryos in the field. Theriogenology 1995; 43: 71-80.
  • Tominaga K. Cryopreservation and sexing of in vivo and in vitro produced bovine embryos for their practical use. J Reprod Develop 2004; 50: 29-38.
  • Utsumi K, Kawamoto T, Kim JH, Iritani A, Sakai A, Komano T. Sex determination of bovine embryos by the polymerase chain reaction using Y spesific primers. J Reprod Develop 1992; 38: 35-43.

Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur

Yıl 2013, Cilt: 10 Sayı: 2, - , 01.06.2013

Öz

Bu çalışmanın konusu, farklı dönemlerdeki in vivo sığır embriyolarının cinsiyetlerinin non elektroforetik PCR yöntemi ile tespit edilerek, cinsiyet dağılımının ve bu dağılımın arasındaki farkların cinsiyetten etkilenip etkilenmediğininbelirlenmesidir. Cinsiyet tayini amacıyla yedi günlük toplam 65 adet in vivo üretilmiş embriyo kullanıldı. Embriyo eldeetmek için rutin süperovulasyon yöntemleri kullanıldı. Tohumlamalardan yedi gün sonra cerrahi olmayan yöntemleuterus yıkaması yapılarak embriyolar toplandı. Elde edilen embriyolar kalite değerlendirmesinin ardından cinsiyetleritespit edildi. Embriyoların trophektoderm kısmından mikro manipülatöre bağlı mikro bıçak ile %10-30 oranında bir parça kesilerek biyopsi materyali alındı. Embriyodan biyopsi alma işlemi %20 FCS içeren D-PBS solüsyonu içerisindeyapıldı. Alınan biyopsi parçaları, lizis ve denaturasyon işlemleri amacıyla PCR cihazına yerleştirildi. Lizisi takiben DNAamplifikasyon işlemi 35 döngü olarak gerçekleştirildi. PCR cihazından çıkarılan tüpler, 302-312 nm dalga boyunda ışıkveren UV lamba altında incelendi. Pembe renk veren embriyo “erkek embriyo” olarak belirlendi. Cinsiyet tayini yapılanembriyolardan kompakt morula aşamasında olan 30 embriyodan 14’ünün erkek (%46.7; 14/30), 16’sının dişi (%53.3;16/30) cinsiyette; blastosist aşamasındaki 35 embriyodan 21’inin erkek (%60.0; 21/35), 14’ünün dişi (%40.0; 14/35)cinsiyete sahip olduğu tespit edildi. Blastosist aşamasındaki erkeklerin oranının (%60.0) morula aşamasına (%46.7)göre daha yüksek bulunmasına rağmen aralarındaki farklılık istatistik önemde bulunmamıştır (P>0.05).

Kaynakça

  • Avery B, Claus BJ, Madison V, Greve T. Morphological development and sex of bovine in vitro fertilized embryos. Mol Reprod Dev 1992; 32: 265-70.
  • Bredbacka P, Veimala R, Peippo J. Survival of biopsied and sexed bovine demi-embryos. Theriogenology 1994; 41: 1023-31.
  • Bredbacka P, Kankaanpaa A, Peippo, J. PCR sexing of bovine embryos: A simplified protocol. Theriogenology 1995; 44: 167-76. Garcia JF, Nogueira MFG, Pupim F, Pantano T, Visintin JA. Pregnancy rates of blastomere biopsied bovine embryos frozen in ethylene glycol. Theriogenology 1997; 47: 268.
  • Hasler JF, Cardey E, Stokes JE, Bredbacka P. Nonelectrophoretic PCR-sexing of bovine embryos in a commercial environment. Theriogenology 2002; 58: 1457-69. Hassun PA, Mello MRB, Porto LPC, Garcia JF. Bovine embryo sexing by primer extension preamplification polymerase chain reaction. Theriogenology 1999; 51: 398. Jafar SI, Flint APF. Sex selection in mammals: A review. Theriogenology 1996; 46: 191-200. King WA, Yadav BR, Xu KP, Picard L, Sirard MA, Supplizi AV, Betteridge KJ. The sex ratios of bovine embryos produced in vivo and in vitro. Theriogenology 1991; 36: 779-88. Kitiyanant Y, Saikhun J, Siriaroonrat B, Pavasuthipaisit K. Sex determination by polymerase chain reaction and karyotyping of bovine embryos at first cleavage in vitro. Scienceasia 2000; 26: 9-13.
  • Kobayashi J, Sekimoto A, Uchida H, Wada T, Sasaki K, Sasada H, Umezu M, Sato E. Rapid detection of male-spesific DNA sequence in bovine embryos using fluorescence in situ hybridization. Mol Reprod Dev 1998; 51: 390
  • Machaty Z, Paldi A, Csaki T, Varga Z, Kiss I, Barandi Z, Vajta G. Biopsy and sex determination by PCR of IVF bovine embryos. J Reprod Fertil 1993; 98: 467-70.
  • Park JH, Lee JH, Choi KM, Joung SY, Kim JY, Chung GM, Jin DI, Im KS. Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenology 2001; 55: 1843-53.
  • Rens W, Welch GR, Johnson LA. Improved flow cytometric sorting of X and Y chromosome bearing sperm: Substantiol increase in yield of sexed semen. Mol Reprod Dev 1999; 52: 50-6.
  • Shea BF. Determining the sex of bovine embryos using polymerase chain reaction results: A six-year retrospective study. Theriogenology 1999; 51: 841-54.
  • Şendağ S, Aydın İ, Çelik HA. İneklerde prenatal embriyonik/fötal cinsiyetin belirlenmesi. Erciyes Üniv Vet Fak Derg 2005; 2(1): 39Thibier M, Nibart M. The sexing of bovine embryos in the field. Theriogenology 1995; 43: 71-80.
  • Tominaga K. Cryopreservation and sexing of in vivo and in vitro produced bovine embryos for their practical use. J Reprod Develop 2004; 50: 29-38.
  • Utsumi K, Kawamoto T, Kim JH, Iritani A, Sakai A, Komano T. Sex determination of bovine embryos by the polymerase chain reaction using Y spesific primers. J Reprod Develop 1992; 38: 35-43.
Toplam 12 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Makaleler
Yazarlar

M. Gürçay Bu kişi benim

M. İssi Bu kişi benim

Y. Gül Bu kişi benim

Yayımlanma Tarihi 1 Haziran 2013
Gönderilme Tarihi 24 Haziran 2014
Yayımlandığı Sayı Yıl 2013 Cilt: 10 Sayı: 2

Kaynak Göster

APA Gürçay, M., İssi, M., & Gül, Y. (2013). Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, 10(2).
AMA Gürçay M, İssi M, Gül Y. Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur. Erciyes Üniv Vet Fak Derg. Haziran 2013;10(2).
Chicago Gürçay, M., M. İssi, ve Y. Gül. “Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 10, sy. 2 (Haziran 2013).
EndNote Gürçay M, İssi M, Gül Y (01 Haziran 2013) Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 10 2
IEEE M. Gürçay, M. İssi, ve Y. Gül, “Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur”, Erciyes Üniv Vet Fak Derg, c. 10, sy. 2, 2013.
ISNAD Gürçay, M. vd. “Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 10/2 (Haziran 2013).
JAMA Gürçay M, İssi M, Gül Y. Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur. Erciyes Üniv Vet Fak Derg. 2013;10.
MLA Gürçay, M. vd. “Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, c. 10, sy. 2, 2013.
Vancouver Gürçay M, İssi M, Gül Y. Investigation of Bovine Viral Diarrhoea Virus in Dairy Cattle Premises Where Aborts Occur. Erciyes Üniv Vet Fak Derg. 2013;10(2).