Published in Scientific Works. C Series. Veterinary Medicine, Vol. LVIII ISSUE 4
Written by Cenariu M., Groza I., Pall Emoke, Parlapan Laura, Ilea Cristina
Embryo biopsy performed in order to obtain a small amount of blastomeres needed for embryo sexing is an invasive method that damages the zona pellucida and therefore decreases the survival capacity of the embryo that is subsequently submitted to cryopreservation. The aim of the present study was to evaluate three biopsy techniques applied in bovine embryos according to their capacity to maintain embryo viability after cryopreservation. Three embryo biopsy techniques (needle, blade and aspiration) were applied on 120 bovine embryos divided into 3 batches (n=40) in order to harvest the blastomeres needed for polymerase chain reaction (PCR) embryo sexing. After the biopsy, the embryos were frozen/thawed using the one step method with ethylene glycol and then transferred into synchronized recipients. DNA was extracted from blastomeres and amplified using bovine Y-chromosome specific primers, in order to determine the sex of the embryo. The pregnancy diagnosis and the assessment of pregnancy rate were performed 30 days after transfer using an ultrasound scanner. There was a significant difference in pregnancy rates according to the biopsy method used: 55% for the needle biopsy, 45% for the aspiration method and 30% for the microblade technique. The accuracy of the PCR sexing method was comparable in all batches, and therefore was not influenced by the biopsy method. The needle method of embryo biopsy proved to be the most suitable as it yielded the highest pregnancy rates and can be successfully applied when harvesting blastomeres for embryo sexing.
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biopsy
