The influence of caffeine supplementation and concerted utilization of enzymatic and mechanical semen liquefaction on freezability of dromedary camel spermatozoa

Abstract

Assisted reproductive technologies have been reported to improve reproductive efficiency and genetic potential in camelids. Two experiments were carried out to determine efficiency of centrifugation in the presence of a mucolytic agent for liquefaction of dromedary semen. In the first experiment, three groups, namely: I. Tris lactose (TL, control), II. Tris lactose supplemented with amylase (TL_A) and III. Tris lactose supplemented with amylase followed by seminal plasma removal via centrifugation (TL_A_Cent.) and re-suspension into enzyme free TL media. After equilibration, control group recorded 62.85 ± 4.28% motility and 1.3 ± 0.13 viscosity score, while TL_A group values were 72.88 ± 3.30% and 0.83 ± 0.07%, respectively. TL_A_Cent. group showed significant viscosity reduction (0.33 ± 0.05) and motility decline 47.85 ± 3.04% with increment in abnormalities and detached acrosome. The second experiment investigated the effect of caffeine addition to tolerate enzymatic and mechanical stress. Using 4 mM caffeine in amylase-treated semen (TL_AC) improved post-thaw motility 50.0 ± 1.29% and recovery rate 77.8 ± 3.83% compared to the control (40.17 ± 2.79% and 62.55 ± 8.39%), respectively. Caffeine supplemented centrifuged samples (TL_AC_Cent.) showed superiority (P < .05) in post-thaw motility and recovery rate (38.33 ± 6.41%, 62.76 ± 8.10%) compared to centrifuged samples TL_A_Cent. without caffeine addition (25.00 ± 2.88% and 40.47 ± 3.48%), respectively. Sperm kinetics showed that TL_A exhibited high (p < .05) values for mostly all sperm kinetics. Caffeine treatments showed superiority in velocity curved line (VCL, µm/s) 94.24 ± 8.44 for TL_AC and 104.25 ± 8.72 for the TL_AC_Cent. group compared to 86.8 ± 5.54 for TL_A, and 85.73 ± 5.99 for the TL_A_Cent. groups. In conclusion, preforming a combined enzymatic-mechanical protocol in the presence of an antioxidant may be crucial for refinement of camel semen cryopreservation.

Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives

“…This is in consonance with previous results in rams [23]. Contrariwise, improved sperm acrosome integrity has been reported after caffeine inclusion in sperm diluent of buffalos [24] and camels [25].…”

Perspectives of nanotechnology in male fertility and sperm function

“…Modern breeding systems require, nowadays, the diffusion of superior genotypes and thus the selection of high quality gametes and embryos. In male reproduction, many factors such as routine procedures of handling and storage, might affect negatively the quality of spermatozoa, and several strategies have been designed to overcome these limitations in order to achieve high in vivo and in vitro fertility rates [21], [22]. Among others, sperm purification and sorting represent a valid tool for the selection of sub-populations of the best spermatozoa (in terms of motility and morphology) within an ejaculate and the discard of defective cells, contaminations and debris.…”

Effect of different calcium concentration on sperm motility and fertilisation capacity of rainbow trout (Oncorhynchus mykiss)

“…It has been reported that Na + ions play an important role in regulation of brook trout spermatozoa activation when Ca 2+‐ free solutions were used (Bondarenko et al., 2014b). On the other hand, supplementing the sperm activator with caffeine has been shown to improve sperm motility in fresh and thawed semen of various species of mammals (Barkay et al., 1977; El‐Bahrawy, 2017; Rota et al., 2018). In rainbow trout, caffeine has increased the time of sperm motility in fresh semen to up to twice the normal value (Valdebenito, 2007).…”