MOLECULAR AND BIOCHEMICAL MARKERS FOR EARLY PREGNANCY DETECTION IN SHEEP

Abstract

The present studY was conducted as in the experimental sheep farm of the !\iuckar Research Center. Egyptian Atomic EnergY Authorit' in association with the Institute nf Graduate Studies and Research. University of .-\lexandria. This study aims to accurateh and economically detect early pregnane usmg techniques of Biotechnnlog'. Consequently. the suggested method may replace techniques used nn"•ada's for pregnane' detection since these techniques have disad,•antages (e.g. long time hel('fe accurateh• determining pregnancy in rectal abdominal palpation and high cost and hazards of using serum progesterone radio immunoassay ( Rl.-\ I 1. The farmer may henetlt from such detection economically by increasing his farm hreeding cycle efticicnc,•. reducing the breeding expenses. and isolating fertile ewes as earl as possible. Thcrel(mreducing abortion incidences. Stillbirths or production of \\Cak lambs.,rhis Slud\\aS rerfonneJ in t\\•o separate expenments.

The first experiment

Twenty four mixed hreed sheep from the experimental sheep of the :\uclear Research Center. Egyptian Atomic Energy Authorit,. were used in this stud'. Blond samples were taken at days 5. 7. 10. 16. 21. and 25 da,•s after mating and a 1\;ati\e-1'.-\GF were applied on the serum of these samples in correlation "•ith serum Pn1gcsteronc Rl.-\ test. In addition. white blood cells "•ere isolated from blood samples tn nhtain purilied total RNA for RT-PCR. Gel images were anal,7.ed using tntallah \.1.11 1!\innlincar Dynamic. Newcastle l 'pontync. t_;K 1 computer snti"arc. The obtained results of this experiment can he summari7.ed in thti1ll,,"ing: I. Serum progesterone RIA ga,•e I 00% accurac' for pregnane' detection. Thi,; result is in agreement with the prcYious recorded studies on this issue. ' The computer software gel image analysis reYealed 7 main fractions in all gels at all days. Protein fractions. expressed in fraction °1o. of pregnant and non-pregnant e\\•es "ere compared. Data anal,•sis sho"ed that se\eral fractions ga,•e a distinguishable difference in fraction°o ,-alue at diff rent da,•s under :<tud'. Fraction 7 re,•ealed ditTerences in aYerage fraction % het\\een pregnant and non­ pregnant ewes at days 7 (36.63 ± 3.33 and -15.82 ::: 2.-13. r specti,•elyl. da' Ill (50.34 ± 1.89 and 56.62 z 2.67. respectiwlyJ and Day 21 (53.22 ± -1.36 and 63.8 =

3.2. respecti,•ely). Fraction -1 revealed ditlerences in aYerage fraction °/o het\\een pregnant and non-pregnant ewes at da' 10123.19 ± 1.81 and 15.85 = 2.23. respectiwlyl. day 16 (7.92 = 0.72 and 5.82:::0.77. respecti,-eJ,•). Day 21 1 16.7R = 2.79 and I 0.2 = 2.06. respectivelY) and l)a, 25 I 8.97 ::: 0.82 and 6.56 ::: 11.83. respectiwlyt. 3. Depending on these results. a douhk blind test "•as applied on 1\\ent' lour e"e' using natiw protein electrophoresis comhined with computer solh••are image analysis at the same days after mating. The obtained results sho\\ed that the accuracy of pregnancy status detection lor subjected ewes using traction % of fraction 7 at day 7 as a marker was 79.19% f(,r pregnancy while the accuracy lor non-pregnancy detection was 87.5%. L:sing traction% ,-alues of tractions 4&7 at day 7& I 0 gave 83.33% accuracy for pregnancy detection and 79.I 7% lor non­ pregnancy detection. Fraction 4 at day I 0 had accuracy of 83.33 % lor pregnancy detection and 91.67% lor non-pregnancy detection.