The aim of this study was to evaluate the viability of bull spermatozoa diluted with commercial semen extender and two culture media stored at controlled room temperature (24 °C) for 72 hours. Two Nguni bulls were used for semen collection with the aid of an electro-ejaculator. After macroscopic evaluation, semen was pooled and aliquoted randomly into Triladyl, modified Ham's F10, and TCM-199 culture media, and then stored at 24 °C. Sperm motility parameters, morphology, and viability were analysed with computer aided sperm analysis (CASA) after 0, 24, 48 and 72 hours. The study was replicated four times, and data were analysed using analysis of variance (ANOVA). Triladyl had significantly higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 hours than modified Ham's F10 (86.8%; 26.5%) and TCM-199 (76.7%; 25.0%) culture media. Ham's F10 had higher progressive motility rate (37.8%) than the other extenders TCM-199 (31.7%) and Triladyl (23.4%). There was no significant difference in viability rate between Ham's F10 (26.5%) and TCM-199 (25.0 %) after 72 hours' storage at 24 °C. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails, between the two Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham's F10 and TCM-199 culture media, stored at 24 °C, and stay viable for 72 hours.
Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.
The present study was undertaken to assess viability of frozen–thawed bull semen collected from the bull’s ejaculate and cauda epididymis. A total of 30 ejaculates were collected from three bulls twice per week for 5 weeks (Control). Caudal epididymis were collected from slaughtered beef cattle of unknown origin from the local abattoir. Caudal epididymal sperm was recovered immediately after slaughtering (EP-0 h) and after cooling at 5°C for 24 h (EP-24). The epididymal and ejaculated samples were each pooled together before being extended with Triladyl. Diluted samples per treatment were loaded into a 0.25-mL French straw and cooled to 5°C in 4 h. Cooled straws were placed 4 cm above liquid nitrogen to freeze for 10 min. Frozen straws were immersed into LN2 and kept for 7 days at −196°C. Samples were analysed immediately after dilution and post-thawing using the computer aided sperm analysis for sperm motility rate, viability and acrosome defects. The highest sperm motility rates were observed with EJ-0 h before and after cryopreservation. However, the difference in sperm motility parameters between EP-0 h and EP-24 h evaluated before and after freezing was not significant (P > 0.05). Furthermore, no significant difference in live cells mean values was observed between the three samples on freezing (P > 0.05). In relation to spermatozoa acrosome defects, there was no significant difference observed among the three samples before and on freezing (P > 0.05). In conclusion, the results from the present study revealed that cooling of epididymides at 5°C for 24 h before the recovery of sperm cells was efficient in preserving epididymal sperm viability. However, ejaculated bull spermatozoa had higher sperm motility and viability rate than epididymal sperm.
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