Supplementation of Thymoquinone Nanoparticles to Semen Extender Boosts Cryotolerance and Fertilizing Ability of Buffalo Bull Spermatozoa

Abstract

Thymoquinone nanoparticles (TQNPs) are broadly utilized in numerous pharmaceutical applications. In the present study, we tested the effects of TQNP supplementation on sperm quality and kinematics, acrosome exocytosis, oxidative biomarkers, apoptosis-like and morphological changes of frozen–thawed buffalo sperm, as well as the fertilizing capacity. Semen was collected from buffalo bulls, diluted (1:10; semen/extender), and divided into five aliquots comprising various concentrations of TQNP 0 (CON), 12.5 (TQNP12.5), 25 (TQNP25), 37.5 (TQNP37.5), and 50 (TQNP50) µg/mL, and then cryopreserved and stored in liquid nitrogen (−196 °C). The results revealed that TQNPs (25 to 50 µg/mL) provided the most optimal results in terms of membrane integrity (p < 0.001) and progressive motility (p < 0.01). In contrast, TQNP50 resulted in a greater post-thawed sperm viability (p = 0.02) compared with other groups. The addition of TQNPs to the extender had no discernible effects on sperm morphology measures. Sperm kinematic motion was significantly improved in the TQNP50 group compared to the control group (p < 0.01). TQNPs effectively reduced the content of H2O2 and MDA levels and improved the total antioxidant capacity of post-thawed extended semen (p < 0.01). The addition of TQNP significantly increased the number of intact acrosomes (p < 0.0001) and decreased the number of exocytosed acrosomes (p < 0.0001). A significant reduction in apoptosis-like changes was observed in TQNP groups. The non-return rates of buffalo cows inseminated with TQNP50-treated spermatozoa were higher than those in the control group (p < 0.05; 88% vs. 72%). These findings suggested that the freezing extender supplemented with TQNPs could effectively enhance the cryotolerance and fertility of buffalo sperm.

Regulatory functions of lecithin nanoparticles on post-thaw quality of buffalo bull semen: a morphological analysis of spermatozoa

“…Chromatin damage was evaluated using an assay described by Agarwal and Said (2003). Flow cytometry was employed to identify apoptotic cells in post-thawed spermatozoa, as described by Khalil et al (2023).…”

“…Assessments of total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels in buffalo spermatozoa during the post-thawing stages of cryopreservation were conducted following the methodologies outlined by Garg et al (2009) and Khalil et al (2023). All analyses were carried out using a spectrophotometer (Spectro UV-Vis Auto, UV-2602; Culver City, CA, USA), and commercial kits were purchased from Biodiagnostic (Giza, Egypt) and used according to the manufacturers' instructions.…”

“…Sperm freezing is vital for the preservation of animal biodiversity, offering a practical approach to mitigate the risk of breed and population extinctions. The utilisation of frozen sperm not only contributes to a reduction in the likelihood of such ex-tinctions but also proves cost-effectiveness in terms of transportation expenses (Khalil et al, 2023). In the context of buffalo sperm, cryopreservation can help lower the costs of feeding large numbers of buffaloes and facilitate the dissemination and monitoring of genetic progress.…”

A Review of the Nutritional Aspects and Composition of the Meat, Liver and Fat of Buffaloes in the Amazon

“…The SFA and MUFA content increases more rapidly with the increase in fat content, as a result of a decrease in the proportion of PUFAs and, consequently, in the PUFA/SFA ratio. Ruminant fat is characterized by having high SFA content and a low PUFA/SFA ratio relative to monogastric animal fat [ 88 , 89 ].…”