Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Animal Collection and Breeding Care
2.3. Sperm Collection, Dilution, and Shipping
2.4. Sperm Concentration and Kinetics
2.5. Sperm Mitochondrial Membrane Potential (MMP) and Intracellular Hydrogen Peroxide (ROS) Content
2.6. Sperm Freezing
2.7. Sperm Packaging, Freezing, and Thawing
2.8. pH and Osmolarity Evaluation
2.9. Biochemical Analyses
2.10. Statistical Analysis
3. Results
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Shipping/Centrifugation Extenders (SCEs) | Osmolarity | pH | ABTS | FRAP | NOx |
---|---|---|---|---|---|
mOsm L−1 | µM | µM | µM | ||
Equiplus | 324 ± 3 A | 6.85 ± 0.05 | 61.1 ± 2.5 A | 163 ± 7 A | n.d. |
BotuSemen | 390 ± 5 B | 6.78 ± 0.03 a | 212.3 ± 4.9 BC | 187 ± 10 A | n.d. |
INRA96 | 317 ± 1 A | 6.91 ± 0.07 b | 32.0 ± 7.6 BD | 236 ± 7 B | n.d. |
Freezing Extenders | |||||
Spectrum Duo Red | 985 ± 5 A | 6.66 ± 0.11 A | n.d. | 1234 ± 10 A | 52.7 ± 2.1 A |
BotuCrio | 1321 ± 7 BC | 6.73 ± 0.13 B | 79.1 ± 4.0 | 1213 ± 15 A | 52.3 ± 3.2 A |
INRAFreeze | 734 ± 3 BDE | 6.85 ± 0.07 BC | n.d. | 379 ± 5 B | 65.3 ± 3.2 B |
HF-20 | 809 ± 3 BDF | 7.02 ± 0.05 BD | n.d. | 360 ± 5 B | n.d. |
ABTS | FRAP | NOx | |
---|---|---|---|
Stallion | µM | µM | µM |
No. 1 | 129 ± 4 | 393 ± 7 | n.d. |
No. 2 | 245 ± 6 | 550 ± 6 | n.d. |
No. 3 | 2202 ± 11 | 405 ± 6 | n.d. |
No. 4 | 28 ± 4 | 556 ± 15 | 64 ± 4 |
No. 5 | 910 ± 11 | 934 ± 18 | 64 ± 6 |
No. 6 | 780 ± 4 | 538 ± 10 | 106 ± 6 |
No. 7 | 305 ± 8 | 259 ± 6 | n.d. |
No. 8 | 673 ± 11 | 489 ± 8 | 350 ± 13 |
No. 9 | 1611 ± 29 | 972 ± 20 | 529 ± 11 |
No. 10 | 708 ± 10 | 301 ± 6 | 54 ± 5 |
Conditioned Shipping/ | ABTS | FRAP | NOx |
---|---|---|---|
Centrifugation Extenders (cSCEs) | µM | µM | µM |
Equiplus | 353 ± 301 | 356 ± 134 | 20 ± 40 |
BotuSemen | 432 ± 344 | 347 ± 167 | 9 ± 29 |
INRA 96 | 443 ± 428 | 415 ± 126 | 36 ± 91 |
Conditioned Freezing Extenders | |||
Spectrum Duo Red | 22 ± 46 A | 930 ± 152 A | 96 ± 154 A |
BotuCrio | 88 ± 167 A | 728 ± 215 BC | 112 ± 99 A |
INRA Freeze | 659 ± 365 B | 388 ± 98 BD | 4 ± 12 B |
HF-20 | n.d. | 704 ± 111 BC | n.d. |
Shipping/Centrifugation Extenders (SCEs) | Freezing Extenders | ||||||
---|---|---|---|---|---|---|---|
Equiplus | BotuSemen | INRA 96 | Spectrum Duo Red | BotuCrio | INRA Freeze | HF-20 | |
TotMot (%) | 78.2 ± 14.4 | 80.0 ± 16.3 | 84.3 ± 16.2 | 28.4 ± 14.8 | 25.7 ± 13.7 | 30.5 ± 13.1 | 25.6 ± 14.9 |
Prog (%) | 23.1 ± 7.0 | 19.6 ± 7.8 | 24.5 ± 5.5 | 11.7 ± 6.6 | 9.6 ± 6.2 | 8.6 ± 5.5 | 7.5 ± 5.1 |
VCL (µm/s) | 95.7 ± 23.3 | 93.1 ± 20.1 | 94.1 ± 18.3 | 65.5 ± 10.9 | 62.8 ± 10.6 | 58.5 ± 7.9 | 59.4 ± 9.5 |
VSL (µm/s) | 35.8 ± 10.2 | 34.9 ± 9.3 | 36.1 ± 8.1 | 34.4 ± 12.1 | 32.2 ± 11.9 | 26.4 ± 7.4 | 27.1 ± 7.7 |
VAP (µm/s) | 59.3 ± 14.8 | 60.3 ±13.7 | 59.8 ±12.3 | 44.7 ± 12.3 a | 41.0 ± 11.3 | 36.1 ± 7.3 b | 36.8 ± 8.1 |
MMP (F0B/F0A) | 34.9 ± 13.6 | 25.6 ± 20.2 | 24.8 ± 21.3 | 8.9 ± 6.8 | 10.1 ± 7.0 | 9.5 ± 6.4 | 9.2 ± 5.6 |
ROS (a.u.) | 24.3 ± 4.3 | 24.1 ± 4.3 | 23.3 ± 2.8 | 12.7 ± 5.6 | 13.3 ± 6.9 | 13.1 ± 6.6 | 13.6 ± 8.8 |
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Boni, R.; Ruggiero, R.; Di Palma, T.; Ferrara, M.A.; Preziosi, G.; Cecchini Gualandi, S. Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production. Animals 2024, 14, 2465. https://doi.org/10.3390/ani14172465
Boni R, Ruggiero R, Di Palma T, Ferrara MA, Preziosi G, Cecchini Gualandi S. Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production. Animals. 2024; 14(17):2465. https://doi.org/10.3390/ani14172465
Chicago/Turabian StyleBoni, Raffaele, Raffaella Ruggiero, Tommaso Di Palma, Maria Antonietta Ferrara, Graziano Preziosi, and Stefano Cecchini Gualandi. 2024. "Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production" Animals 14, no. 17: 2465. https://doi.org/10.3390/ani14172465