Separation of Y-chromosome bearing ram’s sperms using an albumin gradient technique and identification of embryos by PCR

Abstract EN

Several advantages have been suggested for producing sexed sperms including using fewer and genetically superior female animals for replacement.Four hundred active ovaries collected from the slaughter house of Al-shu'alah, the number and type of oocytes, ratios of maturation and fertilization shown that there was a significant difference in the numbers of oocytes (P<0.05) between right and left ovaries.

A high recovery rate was obtained of good oocyte (Grade A) 42.35% (432/1020), fair oocyte (Grade B) 37.54% (383/1020) followed by and poor oocyte (Grade C) 17.84% (182/1020).

There was a significant difference (P<0.05) between the 3 different grades.

grades A and B oocytes, (815/1020) 79.9% of recovered oocytes were cultured.

Maturation rate was 86.38% (704/815).Y- Bearing sperms separation applied by using procedure of the modified albumin technique; either one (8%) or two layers (8 and 16%) of BSA (M1, and M2) at 200, 300 or 400 xg, then used for in vitro fertilization.The in vitro fertilization rate observed was 21.8% (132/604) of matured oocytes by choosing universal primers from sequences that are highly conserved in the X and Y chromosomes, sex-specific sequences were successfully amplified in embryonic lysates.

Bovine serum albumin sexed sperms result in more percentage of male embryos by using one layer of BSA ( 8%) at 200 × g (M1a) and 300× g (M1b) which were 72.7% and 54.5% respectively, and shows a deviation (p<0.05) from the 50% expected percentage for male and female embryos.

While using two layer of BSA (16% and 8% BSA) at the 200 × g (M2a) and at 300× g (M2b) were 81.8% and 63.6% respectively.

When we compare the rate of male embryos produced from IVF by sperms isolated by two layers of BSA (M2a, and M2b), moderate results obtained with M2b (63.6%) while the best results were with M2a separation protocol (81.8%).