Electrofusion between Blastula Cells and Unfertilized Egg in Loach (Paramisgurnus dabryanus)

Electrofusion between blastula cells and unfertilized eggs in loach were investigated usingdielectrophoretic field where, under alternating sinusoidal electric field, blastula cells formed beads-like chain in close contact with the unfertilized egg and cell fusion occurred between eggs and thecells in tight contact with them. The nuclei ofblastula cells were brought into the cytoplasm of therecipient eggs, where they promoted the development of the fused eggs just like the zygote nuclei.But the development of the fused eggs was different from that of zygotes. Several nuclei might enterone and the same egg simultaneously and all of them could undergo division, resulting in severalblastomere after the first cleavage of the recipient egg. Before blastula stage, the embryo developingfrom the fused egg showcd irregular shape, but it was soon regulated and developed to a normalblastula which often continued its development into a normal individual. Cell/egg electrofusion cameto its highest fosion rate (80%) 8nd hatching rate (20%), with cell density at 1×10~3 cells/ml, Ca^(++)concentration at 10 mM, mannitol at 0.2 M and when the blastula cells were digested with 100μg/ml pronase E for 6-10 min at 20℃. The mechanism underlying development of electrofused eggsis discussed. As the result indicates, electrofusion might prove to be a promising biotechnology justas nuclear transplantation.

Differentiating between fertilized and unfertilized eggs prior to incubation based on oxygen flux measurement

One unresolved challenge in the egg industry is how to efficiently and non-invasively detect unfertilized eggs prior to incubation.This detection ability would not only significantly improve hatching rates and reduce costs but also conserve incubator space and prevent poor-quality embryos from contributing to the spread of infections.This study demonstrates a procedure for distinguishing between fertilized and unfertilized eggs prior to incubation by studying the respiratory differences between fertilized and unfertilized eggs using the Non-invasive Micro-test Technique(NMT).A customized micro-testing examination platform,NMT Egg Testing System(NMT-ETS)was constructed for the real-time monitoring of the intensity and rate of oxygen exchange between the egg and its external environment.The results from this study revealed that at room temperature,there is a significant difference in gas exchange rates between fertilized and unfertilized eggs.The results indicate that the oxygen flux of fertilized eggs exceed 20 pmol/(cm^(2)·s),whereas unfertilized eggs show a much lower oxygen flux.Based on the results,the NMT method can be used to effectively distinguish between fertilized and unfertilized chicken eggs.