Post-Thaw Evaluation of Cryopreserved Bull Semen Extended In Four Different Semen Extenders (original) (raw)
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2017
Commercially available OptiXcell® extender was compared with conventional extenders for freezability and in vivo fertility of bull semen. Semen was collected from three Friesian bulls for five weeks (replicate) and qualifying ejaculates (motility >60%, concentration >0.5 billion/mL, volume >1mL) were diluted (37°C; 50 × 106 spermatozoa/ml) with OptiXcell®, tris-citric egg yolk and egg yolk-citrate extenders. Diluted semen was cooled to 4°C in 2 hours, equilibrated for 4 hours and filled in 0.5 ml straws. The straws were kept over liquid nitrogen vapours for 10 minutes and plunged into liquid nitrogen. Percentages of post thaw sperm motility and plasma membrane integrity were recorded higher (P<0.05) in OptiXcell® compared to tris-citric egg yolk followed by egg yolk-citrate extender. Sperm viability (%) were recorded higher (P<0.05) in OptiXcell® compared to tris-citric egg yolk and egg yolk citrate extender. Percentages of normal apical ridge and DNA integrity were h...
Journal of Advanced Biotechnology and Experimental Therapeutics, 2024
Abstract: Cryopreservation has been used extensively for cattle in Bangladesh, however no study was conducted on the cryopreservation of buffalo sperm in Bangladesh. Thus, the current study aimed to evaluate the suitability of different semen extenders for improving the post-thaw quality of Bangladeshi buffalo sperm under a manual cryopreservation protocol. The manual cryopreservation protocol was compared and optimized with a commercial biofreezer protocol. Then, the efficacy of different diluters was evaluated using the optimized manual cryopreservation protocol. Meanwhile, the post-thaw sperm quality in terms of motility and morphology was evaluated by computer assisted sperm analyzer during the optimization process. During manual cryopreservation, the first cooling from 37 °C to 5 °C was done in an equilibration chamber and the second cooling from 5 °C to-120 °C in a Styrofoam box using liquid nitrogen vapor from different distances (0.5, 1.5, 1.6, 2 and 3 inches). Simultaneously, another batch of sperm was cryopreserved using a programmable freezer. The highest number of motile sperms (62.67±1.12; P<0.01) and progressive motility (38.97±1.10; P< 0.001) was observed at 1.6 inches above liquid nitrogen, which were similar to the results obtained from automated biofreezer protocol (65.94±4.65 and 45.54 ± 3.64, respectively). To evaluate the semen extenders' efficacy, one locally developed Tris-fructose-egg yolk-based diluter and two commercial diluters (Andromed, and Triladyl) were used in the freezing of buffalo sperm. The highest recovery and conception rates were observed in sperm diluted with tris-fructoseegg yolk-based (TFE) (82.4% and 80%, respectively). Therefore, it is suggested that this manual cryopreservation protocol and the TFE diluter could be a suitable and inexpensive alternative for Bangladeshi buffalo sperm considering post-thaw sperm quality and fertility.
Animal Reproduction Science, 2008
The process of cryopreservation impairs sperm cell function, potentially leading to a reduction in fertility. The objectives of the present study were to evaluate the effects that cryopreservation using two different extenders has on sperm motility and mitochondrial function, as well as on the integrity of plasma membranes, acrosomal membranes and chromatin, using practical and objective techniques. The focus of the present study was to identify correlations between alterations in sperm membranes and sperm motility in cryopreserved bovine spermatozoa. Seven ejaculates were collected from eight Simmental bulls (n = 56). After collection, semen volume and concentration were assessed for purposes of dilution. Sperm motility was evaluated subjectively and by computer-assisted semen analysis, morphological characteristics were evaluated by differential interference microscopy, the integrity of plasma and acrosomal membranes, as well as mitochondrial function, were determined using a combination of fluorescent probes containing fluorescein isothiocyanate–Pisum sativum agglutinin, propidium iodide or 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide. Chromatin integrity was evaluated using the acridine orange technique. The semen was subsequently divided into two aliquots and diluted with one of two extenders (Bioxcell® or Botu-Bov®), after which both were packaged in 0.5 mL straws and frozen using an automated system. Two straws of semen from each treatment were thawed, and the semen parameters were evaluated as described above. Cryopreservation of sperm reduced motility, damaging plasma and acrosomal membranes, as well as decreasing mitochondrial function. The Botu-Bov® extender was more effective in preserving sperm motility and membrane integrity than was the Bioxcell® extender.