Abstract
The identification of superior sire is largely dependent on the semen quality evaluations, currently through computer-aided sperm class analyser (CASA). Immunoassays present a viable method for analysis of proteins associated with fertility, such as osteopontin. We have targeted osteopontin at an upstream epitope at position 249 - 267 amino acids and a downstream epitope at position 59 - 74 amino acids of full osteopontin protein in raw, BTS, Kobidil, Tris and Citrate liquid preserved Large white and Kolbroek boar semen. The Genetool analysis software was used to quantify the amount of osteopontin detected. Three boars per breed were used and experiments were repeated five times. Our results revealed that the downstream 15 amino acid epitope was more sensitive to osteopontin antibody. Between total motility and osteopontin, Kolbroek boar semen preserved in Kobidil, Citrate, BTS and Tris extenders revealed positive correlations of 0.564, 0.471, -0.960 and 0.330, respectively. Large white boars semen showed correlations of -0.043, 0.655, 0.778, and 0.202 Kobidil, Citrate, BTS and Tris, respectively. For rapid motility and osteopontin, Kolbroek boar semen revealed positive correlations of 0.842, 0.601, 0.164 and 0.202, respectively while large white boars semen showed correlations of -0.909, -0.144, -0.210, and 0.089, respectively. Only BTS and Kobidil showed high negative correlations between osteopontin and viability in Kolbroek (-0.975) and Large white (-0.746) boar semen, respectively. No significant correlations with osteopontin were observed when Tris extender was used in both breeds. In conclusion, Kolbroek boar semen requires osteopontin in Kobodil extender while large white boar semen requires osteopontin in BTS extender.
Highlights
The ultimate aim of any pig breeding programme is to increase the profitability of commercial pork production
Our results revealed that the downstream 15 amino acid epitope was more sensitive to osteopontin antibody
Between total motility and osteopontin, Kolbroek boar semen preserved in Kobidil, Citrate, BTS and Tris extenders revealed positive correlations of 0.564, 0.471, −0.960 and 0.330, respectively
Summary
The ultimate aim of any pig breeding programme is to increase the profitability of commercial pork production. The current semen evaluation techniques, such as CASA, provide a very conservative estimate of their relative fertility [4] [5]. These observations have led to interests in the investigations of fertility markers in boars [3] [6] [7] [8]. The seminal plasma proteins, mainly osteopontin, have been recognized as a potential fertility marker in boars [6] It has been found in the reproductive tract and ejaculated semen, where it modifies semen on route to ejaculation [6] [9]. The down-stream osteopontin epitope at region 59 - 74 with 15 amino acids sequence, and the up-stream epitope at position 249 - 267 with 18 amino acids sequence of full osteopontin protein, provide great targets for osteopontin immunoassays
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