Sperm survival in frozen semen appears to be determined largely by the amount of intracellular enzymes during freezing and thawing. Data is presented to show the effects of different diluents, temperature and glycerol as well as egg yolk concentrations, on glutamic-oxaloacetic-transaminase release. Summary in Chinese and in English

Determination of an extender protocol which will enhance the viability of frozenthawed bovine spermatozoa will allow producers to obtain higher conception rates due to the increased survival rate of the spermatozoa. Ejaculates of six Brangus bulls (age=18 months) were evaluated for spermatozoal motility, acrosomal integrity, and morphological characteristics (collectively called spermatozoal viability) in two experiments to test our hypotheses that (1) the treatment combination of a 4 hr cooling duration and a 2 hr equilibration with glycerol will result in optimum spermatozoal characteristics after freezing and thawing and (2) rank of three selected extenders relative to their effects on spermatozoal viability after freezing and thawing will be egg yolk-citrate (EC), egg yolk-tris (IMV), and skim milk (milk). In experiment 1, an ejaculate from each bull was partially extended and cooled to 4 ºC for either 2 or 4 hr and then allowed to equilibrate with the glycerolated extender for 2, 4, or 6 hr. Spermatozoal viability was assessed at 0, 3, 6, and 9 hr after thawing. In experiment 1, 4 hr of cooling resulted in a higher percentage of motile spermatozoa than did 2 hr of cooling. The 2 hr equilibration with glycerol yielded lower percentages of motile spermatozoa, acrosomal integrity, and morphologically normal spermatozoa than 4 and 6 hr equilibration durations with glycerol. In experiment 2, we observed a decrease in spermatozoal viability for all three extenders upon freezing and thawing. Viability of frozen-thawed spermatozoa extended in the milk was reduced for all incubation durations, and the IMV extender had a higher percentage of motile spermatozoa than the EC extender at 6 hr of incubation. A higher percentage of intact acrosomes was observed with the IMV extender; however, the EC extender had a higher percentage of morphologically normal spermatozoa than the IMV extender. Our results indicate that at cooling duration of 4 hr and a 4 hr equilibration with glycerol provide the highest level of spermatozoal viability post-thaw of the treatments evaluated and that the IMV extender enhances the percentage of spermatozoa with an intact acrosome for frozenthawed spermatozoa over the EC and skim milk extenders.

Cryopreservation of epididymal spermatozoa may be the only opportunity to preserve valuable genetics of males in cases of unforeseen injury or death. Stallion epididymal spermatozoa have been cryopreserved successfully and it has been demonstrated that stallion epididymal spermatozoa are fertile, and pregnancies as well as live foals have been produced. As spermatozoal quality parameters like motility, morphology and viability have a major influence on fertility and pregnancy rates, it is of great interest to describe these and investigate the influence of seminal plasma on these parameters. Fresh and post-thaw parameters (motility, morphology and viability) of stallion epididymal spermatozoa that have been and that have not been exposed to seminal plasma were evaluated, and directly compared to fresh and post-thaw parameters of ejaculated spermatozoa from the same stallions. Six sperm categories of each stallion (n= 4) were evaluated for motility, morphology and viability. These categories were fresh ejaculated spermatozoa (Fr-E), fresh epididymal spermatozoa that had been exposed to seminal plasma (Fr-SP+), fresh epididymal spermatozoa that had never been exposed to seminal plasma (Fr-SP-), frozen-thawed ejaculated spermatozoa (Cr-E), frozen-thawed epididymal spermatozoa that had been exposed to seminal plasma prior to freezing (Cr-SP+) and frozen-thawed epididymal spermatozoa that had never been exposed to seminal plasma (Cr-SP-). Results show that seminal plasma stimulates initial motility of fresh epididymal stallion spermatozoa while this difference in progressive motility is no longer present post-thaw; and that progressive motility of fresh or frozen-thawed ejaculated stallion spermatozoa is not always a good indicator for post-thaw progressive motility of epididymal spermatozoa. This study shows that seminal plasma has a positive influence on the incidence of overall sperm defects, midpiece reflexes and distal cytoplasmic droplets in frozen-thawed stallion epididymal spermatozoa while the occurance of midpiece reflexes is likely to be linked to distal cytoplasmic droplets. Furthermore, we could show that seminal plasma does not have an influence on viability of fresh and frozen-thawed morphologically normal epididymal spermatozoa. In conclusion, we recommend the retrograde flushing technique using seminal plasma as flushing medium to harvest and freeze stallion epididymal spermatozoa. AFRIKAANS : Bevriesing van epididimale spermatozoa mag die enigste geleentheid wees om waardevolle genetika van manlike diere te bewaar in die geval van onverwagse dood of besering. Epididimale spermatozoa van hingste is al suksesvol bevries, is bewys vrugbaar te wees en dragtighede sowel as lewendige vullens is deur die gebruik daarvan gelewer. Aangesien maatstawwe van spermgehalte soos beweeglikheid, morfologie en lewensvatbaarheid ‘n beduidende invloed uitoefen op vrugbaarheid en dragtigheidsyfers, is dit van groot belang om hierdie maatstawwe te omskryf en die invloed van seminale plasma daarop te ondersoek. Vars en ontdooide maatstawwe (beweeglikheid, morfologie en lewensvatbaarheid) van hings epididimale spermatozoa wat of aan seminal plasma blootgestel is of nie, is vergelyk met vars en ontdooide maatstawwe van geejakuleerde spermatozoa. Ses spermkategorieë van elke hings (n=4) is geevalueer vir beweeglikheid, morfologie en lewensvatbaarheid. Die kategorieë was vars geejakuleerde spermatozoa (Fr-E), vars epididimale spermatozoa wat blootgestel is aan seminale plasma (Fr-SP+), vars epididimale spermatozoa sonder blootstelling aan seminale plasma (Fr-SP-), ontdooide geejakuleerde spermatozoa (Cr-E), ontdooide epididimale spermatozoa blootgestel aan seminale plasma voor bevriesing (Cr-SP+) en ontdooide epididimale spermatozoa wat nooit blootgestel is aan seminale plasma nie (Cr-SP-). Uitslae toon dat seminale plasma die aanvanklike beweeglikheid van vars epididimale hings spermatozoa stimuleer, terwyl die verskil in lynbeweeglikheid nie meer teenwoordig is na ontdooiing nie; dat lynbeweeglikheid van vars epididimale spermatozoa of ontdooide geejakuleerde hings spermatozoa nie altyd ‘n goeie aanduiding is vir lynbeweeglikheid na ontdooiing nie. Hierdie studie toon dat seminale plasma ‘n positiewe invloed het op die voorkoms van algehele spermdefekte, midstukreflekse en distale sitoplasmiese druppeltjies in ontdooide hings epididimale spermatozoa terwyl die voorkoms van midstuk reflekse waarskynlik verband hou met distale sitoplasmiese druppletjies. Verder kon ons ook aantoon dat seminale plasma geen invloed het op die lewensvatbaarheid van vars en ontdooide morfologies normale epididimale spermatozoa nie. Ten slotte beveel ons aan dat retrograadse spoeling met seminale plasma as spoelmedium gebruik word wanneer hings epididimale spermatozoa versamel en bevries word. / Dissertation (MSc)--University of Pretoria, 2010. / Production Animal Studies / unrestricted

published_or_final_version / Biochemistry / Doctoral / Doctor of Philosophy

published_or_final_version / Obstetrics and Gynaecology / Master / Master of Philosophy

No description available.

No description available.

Study objectives were to; 1) determine if supplementing of n-3 fatty acids improves membrane integrity, and hence viability and motility of stallion spermatozoa following cold storage, and following cryopreservation, and 2) determine if n-3 supplementation alters the fatty acid composition of stallion spermatozoa. Data indicate that following 90 d of n-3 supplementation daily sperm output and the percentage of morphologically normal sperm in neat semen are increased. Omega-3 supplementation for 90 d did not improve spermatozoal motility or viability following short-term preservation (0, 24 h, 48 h), or following cryopreservation. Although motility was unchanged in this study, individual stallion responses did indicate that n-3 supplementation in stallions with marginal to poor semen quality may benefit from n-3 supplementation. Finally, n-3 fatty acid supplementation does alter plasmalemma fatty acid composition. Spermatozoa from supplemented stallions had increased docosahexaenoic acid (DHA) concentrations as compared to non-supplemented stallions. It is postulated that an increase in long chain n-3 fatty acids, specifically DHA in spermatozoa membrane improves membrane integrity, and thus enhances spermatozoa recovery following the stresses of cold storage and cryopreservation. This phenomenon appears to be beneficial to stallions with marginal to poor quality ejaculates.

No description available.

Ejaculated, density purified, human spermatozoa were exposed to 900 MHz GSM mobile phone radiation at two specific absorption rate levels (SAR 2.0 and 5.7 W/kg) and examined at various time points post exposure. Change in sperm mitochondrial membrane potential was analyzed using flow cytometry. Sperm motility was determined by computer assisted sperm analysis (CASA). There was no effect of 900MHz GSM radiation on mitochondrial membrane potential. This was also the case for all kinematic parameters assessed at SAR of 2.0 W/kg. However, two kinematic parameters (VSL and BCF) were statistically significantly altered after the exposure at SAR 5.7 W/kg. Effects seen cannot be ascribed to heating, as the temperature did not increase by more than 0.3ºC. A thorough investigation at lower SAR levels is required to determine the extent of the influence of RF-EMF on human sperm motility.