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Abstract

Enucleation of the Unfertilized Avian Ovum
An Hoang*, Mike Kjelland, Duane Kraemer
Reproductive Sciences Laboratory, Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX  77843, USAAn Hoang

Abstract

The visual opacity of the avian ovum creates a major obstacle in performing traditional nuclear transfer techniques, specifically, the enucleation of the ovum.  The objective of this study was to explore alternative methods of enucleation using radiation and chemical treatment with Demecolcine.  To determine an effective dose sufficient to inhibit embryonic growth and development, fertilized White Leghorn chicken egg yolks were exposed to varying dosages of gamma radiation (6.4 Gy, 9.2 Gy, and 12.8 Gy).  Similarly, fertilized yolks were separated from their albumin and treated with 0.5μg/ml of Demecolcine for 30 and 60 minute durations before being placed into windowed eggs.  The results showed that the total dosages, as delivered by a Cobalt 60 gamma radiation source, were not sufficient to entirely inhibit embryonic growth.  Also, there was no statistical significance (p>0.05) of the yolks treated with Demecolcine, as compared to the non-treated yolks, to successfully inhibit embryonic growth thus indicating the inability of the current treatment to consistently enucleate the avian ovum.  The present study suggests that much more work needs to be done to elucidate these methods before they can be successfully applied to avian cloning.