Abstract
Simple SummaryThe excessive production of reactive oxygen species (ROS) in cryopreservation and post-thawing affects sperm quality and subsequent fertilizing ability. Antioxidants of natural origin, such as plant extracts, rich in flavonoid and phenolic compounds, are of special interest in scavenging ROS. The supplementation of goat semen extender with 375 µg/mL T. fischeri leaf extract improved the functional and ultrastructural characteristics of cryopreserved sperm by maintaining antioxidant capacity, thus preventing membrane injury and reducing apoptosis.This study evaluated the efficacy of Turraea fischeri leaf extract for maintaining the viability of cryopreserved goat sperm. Ejaculated semen was collected from 5 mature Baladi bucks (50–60 kg, 2–4 years of age) and those samples with mass motility ≥ 70% and sperm concentration ≥ 2.5 × 109/mL were selected, pooled, and divided into 4 aliquots. Each aliquot was diluted in Tris-citric-soybean lecithin extender containing a different concentration of T. fischeri leaf extract (0, 125, 250, or 375 µg/mL). Treated semen samples were cooled to 5 °C, transferred to 0.25-mL French straws, and stored in liquid nitrogen (LN2) at −196 °C. After thawing, membrane integrity was examined by transmission electron microscopy, apoptotic activity by Annexin/propidium iodide staining and flow cytometry, and both enzyme activities and antioxidant capacity by spectroscopic assays. The leaf extract at 375 µg/mL significantly improved semen quality as indicated by enhanced total antioxidant capacity, reduced H2O2 concentration, a greater proportion of structurally intact motile sperm, and concomitant reductions in apoptosis and necrosis. The extract also significantly increased the proportion of sperm with a contiguous plasma membrane and intact acrosome (p < 0.05). Furthermore, LC-MS revealed numerous secondary metabolites in the extract that may contribute to sperm cryopreservation.
Highlights
Artificial insemination (AI) is used widely in agriculture to optimize and spread commercially valuable genetic traits, including in goats
In healthy mammalian sperm cells, total antioxidant capacity (TAC) and reactive oxygen species (ROS) production remain in balance [2], but an increase in ROS production, a decrease in TAC, or both during cryopreservation can result in reduced motility and cell death [3]
The bark extract from the same plant was rich in 20 secondary metabolites belonging to cinchonains and phenylpropanoid-substituted catechin [14]
Summary
Artificial insemination (AI) is used widely in agriculture to optimize and spread commercially valuable genetic traits, including in goats. Cryopreservation can reduce goat sperm quality, motility, and viability, resulting in lower fertility rates [1]. The reduced semen quality results in part from cellular oxidative stress, which causes the peroxidation of unsaturated fatty acids in the biomembrane. In healthy mammalian sperm cells, total antioxidant capacity (TAC) and reactive oxygen species (ROS) production remain in balance [2], but an increase in ROS production, a decrease in TAC, or both during cryopreservation can result in reduced motility and cell death [3]. Exogenous antioxidants are frequently added to cryopreservative solutions to sustain semen quality prior to AI [6]. Several studies have confirmed ROS scavenging capacity of various medicinal plant extracts to improve sperm motility and increase fertility rates [7,8,9,10]
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