Influence of alpha linolenic acid on the motility, viability, antioxidant activity and fertility of frozen-thawed New Zealand white rabbit buck semen

[EN] Freezing and thawing processes result in production and accumulation of high concentrations of reactive oxygen species that are detrimental to spermatozoal motility and fertility. Therefore, supplementation of exogenous source of antioxidants to freezing diluent is crucial. The aim of the present study was to investigate for the first time whether supplementation of semen diluent with alpha linolenic acid (ALA) can improve motility, viability, membrane integrity, antioxidant status and fertility of post-thaw rabbit spermatozoa. Semen was collected and pooled from fifteen New Zealand white rabbit bucks. Semen samples were diluted with a tris-citrate-glucose (TCG) extender supplemented with ALA (0, 50, 75 and 100 μmol). Then, extended rabbit semen was cooled at 5°C and cryopreserved in liquid nitrogen. After thawing, spermatozoal quality parameters (individual motility %, viability %, osmotic resistance %, and acrosome integrity %), antioxidant activity (SOD, CAT, and GSH activities), lipid peroxidation (malondialdehyde) and fertility (conception and kindling rates) were evaluated. Results revealed that supplementation of rabbit semen extender with 50 μmol ALA significantly (P<0.05) increased spermatozoal characteristics including motility (56.54%), viability (60.01%), acrosome status (72.66%) and membrane integrity (59.13%). The activity of semen antioxidant enzymes (SOD, CAT, and GSH) showed a significant improvement with a marked decrease in lipid peroxidation. Moreover, the conception (73.30%) and kindling (70.00%) rates were significantly (P<0.05) higher in does inseminated with thawed semen treated with 50 μmol ALA in comparison with other concentrations (0, 75 and 100 μmol). In summary, supplementation of rabbit semen extender with 50 μmol ALA improved motility, viability, membrane integrity, acrosome integrity, antioxidant enzymes activity and fertility of post-thaw rabbit spermatozoa. Our findings suggested that higher concentrations of ALA are detrimental to post-thaw characteristics of New Zealand white rabbit buck spermatozoa. To achieve better results, the semen freezing extender should be supplemented with ALA at lower concentrations, especially 50 μmol.El-Shahat, KH.; Fadl, AM.; Abdelnaby, EA. (2022). Influence of alpha linolenic acid on the motility, viability, antioxidant activity and fertility of frozen-thawed New Zealand white rabbit buck semen. World Rabbit Science. 30(3):219-226. https://doi.org/10.4995/wrs.2022.1704221922630

Quality assessment of cryopreserved New Zealand white rabbit spermatozoa in INRA-82 extender containing different cryoprotectants

[EN] The present study aimed to evaluate the effect of supplementation of INRA-82 semen extender with different cryoprotectants (dimethyl sulphoxide; DMSO vs. dimethyl formamide; DMF) on the quality of white New Zealand rabbit buck spermatozoa. We also investigated the possible association between the synergistic action of DMSO and DMF and their relation with INRA-82 extender composition. Semen was collected and pooled from 8 adult rabbit bucks. Pooled semen samples were diluted 1:1 with INRA-82 extender supplemented with DMSO 8%, DMF 8% or a combination of DMSO 4% and DMF 4%. The diluted semen samples were cryopreserved in 0.25 plastic straws. After thawing, progressive motility, sperm viability, sperm abnormalities, membrane integrity, acrosome status, viability index and DNA integrity were evaluated. The results showed that dilution of rabbit buck semen in INRA-82 supplemented with DMSO and DMF (4% each) before freezing significantly (P<0.05) improved sperm motility (42.00%), percentage of live spermatozoa (45.30%), proportions of spermatozoa with intact acrosome (59.75%) and percentage of spermatozoa with non-fragmented DNA (86.04%), compared to those diluted in INRA-82 supplemented either with DMSO 8% (+9, +10, +5 and +7 percentage points, respectively) or with DMF 8% alone (+18 +18, +12 and +9 percentage points, respectively). In conclusion, dilution of rabbit buck semen before freezing with INRA-82 extender supplemented with a combination of DMSO 4% and DMF 4% improved quality of frozen-thawed New Zealand White rabbit spermatozoa. Furthermore, our results also suggest that supplementation of INRA-82 with DMSO or with DMF alone at higher concentrations deteriorates the sperm quality.Fadl, AM.; Ghallab, AM.; Abou-Ahmed, MM. (2019). 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Eco-friendly Synthesized Zinc Oxide Nanoparticles Improved Frozen-thawed Semen Quality and Antioxidant Capacity of Rams

Zinc oxide nanoparticles (ZnO-NPs) have antimicrobial, antioxidants, and anticancer properties. This study aimed to improve the post-thawing semen characteristics of rams using three concentrations of ZnO-NPs. ZnO-NPs in 0.0, 0.5, 1.0, 1.5 mg/ml tris-based diluents were used to extend semen collected from five Awassi rams twice weekly for two months. Post-thaw semen evaluation and measurement of Malondialdehyde (MDA), nitric oxide (NO), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), total cholesterol (CHO), low density lipoproteins (LDL), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), zinc, and copper (Cu). With increasing ZnO-NPs concentrations, the post-thaw sperm total motility (STM %), progressive motility (SPM) %, viability (SV%), functional membrane integrity (FMI %), acrosome integrity (AI %), NO, LDH, ALP, Cu, and zinc ascended linearly (P≤0.0001) but the abnormal sperm (AS%) and SOD decreased (P≤0.0001). The addition of 1.0 mg/ml ZnO-NPs increased (P≤0.0001) GPx but decreased (P≤0.0001) CHO and LDL. All concentrations of ZnO-NPs preserved (P≤0.0001) high AA compared to 0.0mg/ml and did not influence MDA. The addition of 0.5 mg/ml ZnO-NPs reduced (P≤0.059) Cu concentrations. All semen characteristics had strong positive correlations with zinc, LDH, and ALP and negative correlations with SOD. AS % had negative correlations with zinc, ALP, and LDH and positive correlation with SOD. All the three added concentrations of ZnO-NPs improved the post-thaw semen motility, viability, fertilizing capacity, and antioxidant capacity but 1.5mg/ml semen diluents proved the best concentration for semen preservation

Methanolic pomegranate dried peel extract improves cryopreserved semen quality and antioxidant capacity of rams

Objective: To select the appropriate concentrations of methanolic pomegranate extract supplemented in rams' semen extender for obtaining the best-cryopreserved semen quality. Methods: Tris-based semen extender was supplemented with 0.0, 0.40, 0.48, and 0.56 mg/mL pomegranate peel methanolic extract to extend semen collected from five native rams twice weekly for two months (n=80). Pooled (n=16) post-thaw semen characteristics were determined. Thawed seminal plasma of all supplemented and control groups were used to measure malondialdehyde (MDA), nitric oxide (NO), glutathione peroxidase (GPx), superoxide dismutase (SOD), ascorbic acid, zinc, copper, total cholesterol, low-density lipoproteins (LDL), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP). Results: The supplementation of Tris-based semen extender with 0.48 mg/mL semen extender resulted in the highest post-thaw sperm total motility (P0.05), and LDH (P>0.05). Conclusions: Pomegranate peel methanolic extract 0.48 mg/mL supplemented to Tris-based semen extender of rams is the best enrichment in preserving the sperm post-thaw characteristics via improving biochemical profiles and antioxidant capacity