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.

Portulaca oleracea contains a wide range of chemical constituents, including flavonoids, alkaloids, polysaccharides, fatty acids, terpenoids, sterols, proteins, vitamins, and minerals. These chemical constituents are linked to the plant’s diverse pharmacological properties, such as antimicrobial and antioxidant activities. Therefore, this study is aimed at evaluating the effect of adding two concentrations (50 or 100 μg/mL) of Portulaca oleracea leaf ethanolic extracts in a native form (POLE) or nanoformulation (POLENF) on the antioxidant status of sperm medium to improve goat sperm freezability. Semen was evaluated postequilibration (5°C for 2 h), postthawing, and 2 h postincubation (37°C and 5% CO₂). Phytochemical analysis showed the presence of 30 compounds in POLE, including phenolic acids and flavonoids and their glucosides. Total flavonoids were higher in POLE than in POLENF. Both extracts showed strong free radical scavenging capacity by the 2,2‐diphenyl‐1‐picryl‐hydrazyl‐hydrate assay, which was higher for POLENF than POLE, whereas the antioxidant activity by the ferric reducing antioxidant power assay had no differences. In comparison to free extender, the addition of POLE (100 μg) or POLENF (50 and 100 μg) significantly improved sperm motility and viability postequilibration and postincubation and antioxidant status of sperm medium postthawing. All POLE and POLENF concentrations significantly improved motility, viability, membrane integrity, and apoptosis of spermatozoa postthawing. Enzyme activities of AST, ALT, and LDH in sperm medium postthawing were not affected by treatments. In comparing different types and concentrations of POLE with free extender, the addition of POLENF (100 μg/mL) to goat semen extender gave the most valuable results as an effective free radical scavenger and antiapoptotic during cryopreservation.

The aim of the current study was to assess the impacts of aqueous extract of Moringa oleifera leaves (MOLE) on semen quality and sperm freezing ability of bucks. 5 sexually mature Damascus bucks (65-75 kg live body weight, 2-4 years of age) were used for collecting semen once per week for seven weeks. Only semen with a mass motility of ≥ 70% was pooled. Pooled semen had been divided into 4 aliquots, with the free extender (C=control, no MOLE) and the extenders supplemented with MOLE at levels of (10, 20, and 30 µl/mL) as an antioxidant. Tris-citric acid-extender was used to dilute semen at a rate of 1:20 using different MOLE levels, then chilled to 5 °C for two hours (equilibration period), placed into 0.25 mL straws and finally kept in liquid nitrogen at -196 oC for two weeks. After dilution, equilibration, and thawing, semen was assessed visually and via CASA. The results showed that MOLE, at a level of 30 µl/ml, had a beneficial effect on all sperm characteristics, all types of sperm motility, and normal forms assisted by CASA in post-diluted semen, after equilibration and thawing. The highest MOLE level (30 µl/ml) considerably improved the sperm kinetic indexes and all sperm velocity types (VCL, VSL, and VAP).In conclusion, Moringa oleifera leaves (30 µl) in semen extender of bucks is good tool for improving semen quality and sperm freezing ability of bucks

Green tea extract is plant–derived natural antioxidant. The purpose of this study was to investigate the effect of the supplementing semen extender (Tris-citric-soybean lecithin) with different levels of aqueous (AGTE) or methanolic (MGTE) green tea extract (0.5, 1.0 and 1.5%) on the motility, livability, normality, plasma membrane integrity, and kinetic parameters of ram spermatozoa in diluted and equilibrated semen. Semen was collected from 5 rams once a week for 7 weeks using an artificial vagina. Results showed positive effect of extender containing 1% MGTE on visual motility, livability, abnormality, membrane integrity, rapid motility, sperm velocity, and normal form in post-equilibrated semen. It is possible to infer that the inclusion of methanolic green tea extract at a level of 1% in semen extender had a substantial effect on sperm quality, normality, function, and velocity of ram spermatozoa in Rahmani semen after dilution and equilibration for 4 h at 5oC.

The aim of this study was to evaluate the impact of supplementing Tris-extender with different levels of nano-oleic acid (NOA) on sperm characteristics, and enzyme activity and antioxidant status of post-thawed sperm medium of bull semen. Semen was collected by artificial vagina from 5 Holstein bulls, then pooled, diluted by Tris-egg yolk extender supplemented with NOA at levels of (0, 1, 2, 4 and 6 µL/mL), equilibrated at 5°C for 4h, loaded in straws, and stored in liquid nitrogen, then thawed at 37oC for 30 s for evalua-tion. Supplementing Tris-extender with nano-oleic acid (5 µL) increased (P<0.05) the per-centages of progressive motility and membrane integrity after dilution, progressive motili-ty, livability, and membrane integrity after equilibration, as well as increasing progressive motility, livability, and membrane integrity, and decreasing abnormality of sperm cells after thawing. Also, recovery rate of progressive motility, livability, and membrane integ-rity after equilibration and thawing was the highest for semen supplemented with 1µL nano-oleic acid. Supplementing Tris-extender with nano-oleic acid at a level of 5 or 2µL decreased (P<0.05) AST and ALT activities, and increased (P<0.05) TAC level and GPx activity as compared to free-extender. Pregnancy rate of cows inseminated with 1µL nano-oleic acid showed higher (P<0.05) pregnancy rate (57.5%) than in free-extender (32.5%) or other nano-oleic acid levels (25-42.5%). In conclusion, addition of 1µL NOA in Tris-extender of bull semen can be used as a strategy for improving freezing ability and fertilizing capacity of cryopreserved bull sper-matozoa.