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A High-Throughput Method For Zebrafish Sperm Cryopreservation and In Vitro Fertilization
Published on: July 6, 2009
Inseminating fresh or cryopreserved semen for maximum efficiency: implications for gene banks and industry
H D Blackburn1, F Silversides, P H Purdy
1National Animal Germplasm Program, National Center for Genetic Resources Preservation, Agricultural Research Service, USDA, 1111 S. Mason St., Fort Collins, CO 80521, USA. Harvey.blackburn@ars.usda.gov
Abstract:
Developing gene bank germplasm collections for animal genetic resources requires establishing germplasm collection goals, that consider capturing the genetic diversity of the population in question and the amount of germplasm required for its reconstitution or other purposes, or both. Computing collection goals for chickens is complicated, compared with mammalian species, due to the multiple chances a single insemination of semen has to fertilize an egg. To address this issue, fertility data were used in conjunction with econometric procedures for determining production efficiency and diminishing returns. Experimental treatments consisted of inseminating fresh semen intravaginally (FIV), frozen-thawed semen inseminated intramagnally (FTIM), and frozen-thawed semen inseminated intravaginally (FTIV). Analysis revealed that the maximum efficiency for a single insemination was at postinsemination d 6, 8, and 3 for FIV, FTIM, and FTIV, respectively. But, additional benefit from a single insemination can be garnered by continuing to collect and incubate eggs to d 11, 17, and 11 for FIV, FTIM, and FTIV, respectively. By extending the insemination interval, the number of fertile eggs can be increased by 62 (FIV), 62 (FTIM), and 48% (FTIV). The ramifications of these results are profound when placed in the context of germplasm collection for gene banks. By using the FTIM treatment, the number of germplasm samples needed to secure a chicken breed, at the 150% level, can be reduced from the FAO projection of 2,454 to 386 straws (0.5 mL). Such a change represents a substantial reduction in collection, processing, and storage costs for gene banks. For industry, the results suggest that extending the time interval between inseminations will yield more fertile eggs and create opportunities to increase the number of hens mated to a rooster.

