Effects of biopsy volume on the fidelity in sexing of bovine embryos by PCR and the freezability of biopsied embryos
1996
Kasai, K. (Osaka-fu. Agricultural and Forestry Research Center, Habikino (Japan)) | Fujitani, Y. | Otani, S.
For a successful sex determination of bovine embryos using Polymerase chain Reaction (PCR), the effects of the difference of biopsy volume on the accuracy in detection of the male-specific signal and the freezability of damaged embryos by biopsy were investigated. In experiment 1, 50 or 30% volume of bovine embryos at the expanded blastocyst stage were biopsied. PCR trials were performed using the biopsy (portion T) and the rest part of the embryo (portion I) as the template, respectively. The PCR results were compared between the two portions of each embryo to assess the reliability of the results from PCR using portion T. When 50% volume of the embryos was biopsied, the results from PCR using portion T were agreed with those from portion I in a high proportion of 93.3%. But when 30% volume of the embryos was biopsied, the results from PCR using portion T were agreed with those from portion I in a proportion of 75.0%. This indicates that our present PCR program requires more than 30% volume of an embryo at the expanded blastocyst stage as the template. In Experiment 2, viability and the quality of biopsied embryos after freeze-thawing were examined. In vitro survival rates 24 hr. after thawing was almost equal in 30%-biopsied embryos, 50%-biopsied embryos and intact embryos. However, the proportion of Grade A embryos in survived 50%-biopsied embryos was significantly (P0.01) lower than that in other two groups. These results suggest that the biopsy in which 30% of the entire volume is removed does not compromise the freezability of embryos at the expanded blastocyst stage and that the further study is needed to improve the sensitivity of our PCR program
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