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Attempts to promote the use of cryopreserved bovine semen: Effect of prostaglandin F2-alpha, sucrose and short-term dry ice storage

dc.contributor.advisorGauly, Matthias Prof. Dr. Dr.
dc.contributor.authorAbdussamad, Abdussamad Muhammad
dc.date.accessioned2014-01-09T10:12:26Z
dc.date.available2014-01-09T10:12:26Z
dc.date.issued2014-01-09
dc.identifier.urihttp://hdl.handle.net/11858/00-1735-0000-0022-5DEE-1
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-4309
dc.language.isoengde
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subject.ddc630de
dc.titleAttempts to promote the use of cryopreserved bovine semen: Effect of prostaglandin F2-alpha, sucrose and short-term dry ice storagede
dc.typedoctoralThesisde
dc.contributor.refereeHoltz, Wolfgang Prof. Dr.
dc.date.examination2013-10-30
dc.description.abstractengThis dissertation argues that improvement of conception to artificial insemination (AI) through approaches that enhance post-thaw quality of bull semen might sustain enthusiasm for this technology and present a greater chance of capturing superior genetics from proven AI sires; thus, promoting the use of cryopreserved bovine semen. The general objective of this work is to determine the efficacy of some strategies aimed at improving post-thaw motility of bull spermatozoa. The general objective was, therefore, divided into three specific objectives: 1. Investigate the effect of PGF2-alpha addition after thawing on viability of bovine spermatozoa. In that context, extenders with or without egg yolk were compared to establish extender most suitable for PGF2-alpha supplementation. 2. Determine the effect of incorporation of sucrose alone or in combination with glycerol into customized (Tris-egg yolk) and two commercial (Steridyl® and AndroMed®) extenders on post-thaw motility of bovine spermatozoa. 3. Assess the effect of temporary storage of bovine semen originally stored in liquid nitrogen on dry ice and of refreezing of thawed semen on spermatozoon motility. The above-mentioned specific objectives were analysed in Chapters 2 to 4. Chapter 2 is entitled, "In vitro effect of type of extender and addition of prostaglandin F2-alpha post-thawing on the motility of bovine spermatozoa" and it addresses the first specific objective. Results of Experiment 1 show that percent total motility decreased significantly (p<0.05) as bull semen passed through different stages of cryopreservation from extension (65 ± 1%) via equilibration (60 ± 1%) to 15 min (49 ± 3%) and 7 days (46 ± 2%) of immersion in liquid nitrogen (LN2). The difference between storage times (15 min or 7 days) in LN2 was not significant. Post-thaw motility in AndroMed® extender was significantly (p<0.05) higher (59 ± 2%) than in both TriladylTM and Tris-egg yolk-glycerol extenders (both 53 ± 2%). No effect of individual bull was observed (p>0.05). In Experiment 2, results revealed that there was no significant (p>0.05) effect of PGF2-alpha concentration on spermatozoon motility. However, a slight but significant (p<0.05) effect of individual bull was recorded. In conclusion, frozen-thawed bull spermatozoa were capable of tolerating PGF2-alpha up to a concentration of 30% (v/v) in AndroMed® without adverse effect on total motility. Further studies should be attempted to test the effect of prostaglandin F2-alpha added to semen after thawing on success rates at insemination. Chapter 3 is concerned with the second specific objective and it is captioned "Sugar supplementation in customized and commercial extenders: The use of sucrose solely and in combination with glycerol". Results show that relative to Tris-egg yolk containing 6.8% glycerol without sucrose (control), semen in same extender containing 150 mM sucrose and 3.4% glycerol had a relative motility of 68 (SE 3) % which decreased to 52 (SE 3) % in extender with 300 mM sucrose and devoid of glycerol. In semen diluted in Steridyl® with 150 mM or 300 mM sucrose, relative motility was significantly decreased to 67 (SE 3) % in the former and 31 (SE 4) % in the latter. In AndroMed® extender with 150 mM sucrose, motility had significantly decreased to 70 (SE 5) % and with 300 mM sucrose to 42 (SE 6) %. Tris-egg yolk extender containing 150 mM sucrose with 3.4% glycerol proved to be better than the same extender containing 300 mM sucrose without glycerol. Motility in Steridyl® and AndroMed® extenders containing 150 mM sucrose was better than that of the same extenders with 300 mM sucrose. However, motility in Tris-egg yolk extender containing 300 mM sucrose without glycerol was better than that of commercial extenders containing glycerol in combination with 300 mM sucrose. Efforts were made to employ an extender devoid of glycerol but it was observed that a small amount of sucrose in combination with glycerol proved to be advantageous. Complete replacement of glycerol with sucrose in the customized Tris-egg yolk extender retained some motility though not as good as same extender with 150 mM sucrose and 3.4% glycerol. It, therefore, appears that sucrose could be used in semen extenders in order to reduce, partly or completely, the amount of glycerol added to such extenders; thus, ameliorating the toxicity of glycerol when used in high concentrations. Chapter 4 deals with the last specific objective and is entitled "Temporary storage on dry ice of bovine semen originally stored in liquid nitrogen and the effect of refreezing". Results show that neither a 1h nor a 6h sojourn on dry ice affected spermatozoon motility regardless whether semen was thawed immediately or after being returned to LN2 (p>0.05). Intensity of progressive motility was virtually unimpaired by the respective treatments. Post treatment motility rates were reduced by a factor of 10 as compared to semen not subjected to refreezing. No significant difference in percent post-thaw motility after refreezing was observed between semen samples that had been transiently stored on dry ice for 0, 1 or 6 hours (p>0.05). Semen that underwent refreezing on dry ice rather than in LN2 vapour before being returned to LN2 exhibited a significantly higher post-thaw motility rate (p<0.05). Although the proportion of motile spermatozoa was drastically reduced, the intensity of progressive forward motion was satisfactory. Temporary dry ice storage appears to have no adverse effect on percent motility of bovine spermatozoa; thus, its use as a viable option for transport of frozen semen should be explored provided in vivo fertility is proven.de
dc.contributor.coRefereeKnorr, Christoph Prof. Dr.
dc.subject.engSemen cryopreservation; spermatozoon motility; bovine semen; prostaglandin F2-alpha; semen extender; sucrose; glycerol; semen storage; dry ice; liquid nitrogen; semen refreezingde
dc.identifier.urnurn:nbn:de:gbv:7-11858/00-1735-0000-0022-5DEE-1-6
dc.affiliation.instituteFakultät für Agrarwissenschaftende
dc.subject.gokfullLand- und Forstwirtschaft (PPN621302791)de
dc.identifier.ppn775976881


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