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Effects Of Cryopreservation And Constituents Of Semen Extenders On Mitochondrial Function Of Bull Spermatozoa, Abdulhakeem Hashim Eljarah
Effects Of Cryopreservation And Constituents Of Semen Extenders On Mitochondrial Function Of Bull Spermatozoa, Abdulhakeem Hashim Eljarah
LSU Doctoral Dissertations
This study investigated the effects of semen extender constituents and cryopreservation on bovine spermatozoal mitochondrial function. Three yearling Holstein bulls were used. Two ejaculates per bull were collected and pooled on a weekly basis for five weeks and extended in four treatments: 1) sodium citrate egg yolk extender with antibiotics (lincomycin, spectinomycin, gentamicin and tylosin); 2) ¡°1¡± with glycerol; 3) ¡°2¡± without antibiotics; and 4) ¡°1¡± without antibiotics. Each was divided into portions for analyses before freezing and after cryopreservation. The pre-freeze and thawed semen were transferred to a 37¡ãC water bath, the same assays were performed. In experiment 1, …
Gamete And Cell Technologies For Use In Biological Resource Banking, Liesl Nel-Themaat
Gamete And Cell Technologies For Use In Biological Resource Banking, Liesl Nel-Themaat
LSU Doctoral Dissertations
Biological resource banking is becoming important for endangered species conservation. A series of experiments were conducted to address issues concerning collection and utilization of biomaterials from Gulf Coast Native (GCN) sheep (model species) (Ovis aries) and Eland antelope (Taurotragus oryx). In the first experiment, two ejaculates were collected 10 minutes apart from each of five rams three times a week for three weeks to maximize output and minimize handling time. Semen volume, concentration and total number of spermatozoa were significantly greater in first ejaculates, whereas pre-cooled, cooled and post-thaw motility, as well as sperm survival, were greater in second ejaculates. …
Cryopreservation And Intracytoplasmic Sperm Injection With Bovine Epididymal Spermatozoa, Carlos Andres Guerrero
Cryopreservation And Intracytoplasmic Sperm Injection With Bovine Epididymal Spermatozoa, Carlos Andres Guerrero
LSU Doctoral Dissertations
Recently, interest in the preservation of epididymal sperm as a potential source of valuable genes for genome resource banks has escalated. The development of a successful protocol to recover and cryopreserve sperm harvested from the epididymides would salvage germplasm from genetically valuable males that are injured and can no longer mate or have unexpectedly died and can be used as a model for the preservation of male gametes from endangered species. In a series of experiments, epididymal sperm was successfully harvested, cryopreserved and used for intracytoplasmic sperm injection. In Experiment I, ethylene glycol was found to cause significantly (P<0.05) less osmotic damage to bovine sperm during a one step addition and/or removal at 4°C as compared with glycerol in all concentrations evaluated. Furthermore, prolonged exposure (5 days at 4°C) of ethylene glycol was found to be less toxic than glycerol to sperm. In Experiment II, it was demonstrated that glycerol was more effective than ethylene glycol in providing protection against freezing injury during the cryopreservation process in the concentrations evaluated. In Experiment III, it was demonstrated that epididymal sperm retrieval using seminal plasma is beneficial to enhance sperm overall and progressive motility characteristics and to protect it from morphological abnormalities derived from the freezing process. In Experiment IV, a one step dilution process for removal of glycerol from cryopreserved epididymal sperm was found to significantly affect plasma membrane integrity and mitochondrial function of sperm previously exposed to seminal plasma. However, seminal plasma exposure did not have any significant detrimental effect on acrosome integrity. Furthermore, it was demonstrated that the longevity and survivability in vitro during a 4-hour incubation period at 37°C of post-thaw epididymal sperm exposed to seminal plasma prior to cryopreservation was not compromised when compared with the control extended sperm. In Experiment V, we have demonstrated that fertilization, blastocyst and fetal development could be achieved with cryopreserved bovine epididymal sperm by intracytoplasmic sperm injection (ICSI). To our knowledge, this is the first report in the United States and second in the world to use bovine epididymal sperm for ICSI. We achieved far markedly improved blastocyst rates over those results recently reported in the first study originating in Japan.
Preservation Of Sperm Harvested From The Rat, Caprine, Equine, And Bovine Epididymis, Aida Nioma James
Preservation Of Sperm Harvested From The Rat, Caprine, Equine, And Bovine Epididymis, Aida Nioma James
LSU Doctoral Dissertations
The interest in preserving endangered species has increased the amount of attention lent to the recovery of functional sperm from the epididymides of deceased males (Foote, 2000). Postmortem specimens have a finite time period before decomposition affects functionality. Determination of this “window of opportunity” to harvest and preserve epididymal sperm would be beneficial for further research in sperm preservation and assisted reproductive technologies. The objective of these studies was to determine 1) the “window of opportunity” to collect viable rat, caprine, equine and bovine epididymal sperm, 2) if epididymal sperm collected could be cryopreserved, 3) to test two common cryoprotectants …