Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Animals
2.2. Semen Collection
2.3. Experimental Design
2.4. Fresh Ejaculate Evaluation
2.5. Semen Centrifugation
2.6. After-Centrifugation Sperm Evaluation
2.7. Freeze-Test
- -
- Lac-EDTA: straws were frozen directly without a cooling phase [28].
- -
- Inra-Freeze®: straws were frozen after being slowly cooled to 4 °C (−0.3 °C/min) over an hour, as recommended by the manufacturer.
- -
- BotuCrio®: straws were frozen after being cooled at 4 to 6 °C for 20 min, as recommended by the manufacturer.
2.8. Post-Thaw Sperm Evaluation
2.9. Statistical Analysis
3. Results
3.1. Prior Cryopreservation Evaluation
3.2. Post-Thaw Evaluation
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Stallion | Age (Years) | Ejaculate | Group | Fresh | After Centrifugation Protocols | ||
---|---|---|---|---|---|---|---|
S | Protocol 1 CC | Protocol 2 CC | |||||
1 | SC | 76.9 | 67.0 | - | - | ||
A | 9 | 2 | SC | 70.8 | 59.2 | - | - |
3 | SC | 68.4 | 67.9 | - | - | ||
1 | SC | 69 | 69.4 | - | - | ||
B | 17 | 2 | NP | 44.6 | - | 33.1 | 42.6 |
3 | NP | 42.1 | - | 49.2 | 50.7 | ||
1 | SC | 67.2 | 59.3 | - | - | ||
C | 15 | 2 | NP | 38.1 | - | 12.4 | 24.7 |
3 | AACC1-2 | 56.4 | - | 75.7 | 58.4 | ||
D | 9 | 1 | AACC1 | 41.9 | - | 65.4 | 53.6 |
2 | AACC1 | 41.9 | - | 65.4 | 53.6 | ||
1 | AACC2 | 56.5 | - | 54.3 | 66.8 | ||
E | 19 | 2 | NP | 49.6 | - | 49.0 | 48.5 |
3 | NP | 49.7 | - | 41.8 | 32.0 | ||
1 | SC | 71.2 | 52.3 | - | - | ||
F | 7 | 2 | AACC1 | 46.3 | - | 61.8 | 49.8 |
3 | AACC1-2 | 52.1 | - | 60.8 | 64.3 |
GROUP | SC | AACC1 | AACC2 |
---|---|---|---|
MOT5 | 59.10 a ± 20.88 | 51.55 a ± 14.88 | 44.22 a ± 15.11 |
PMOT5 | 45.59 a ± 17.31 | 40.29 a ± 17.09 | 38.51 a ± 12.82 |
VCL5 | 84.08 a ± 14.61 | 76.54 a ± 12.94 | 81.01 a ± 11.69 |
VSL5 | 59.71 a ± 13.67 | 62.43 a ± 10.54 | 68.88 a ± 7.45 |
VAP5 | 69.02 a ± 13.39 | 68.93 a ± 11.45 | 73.39 a ± 8.50 |
LIN5 | 71.99 b ± 8.68 | 81.87 a ± 6.55 | 84.54 a ± 6.82 |
STR5 | 88.89 a ± 5.54 | 90.75 a ± 5.56 | 93.66 a ± 3.10 |
ALH5 | 2.58 a ± 0.54 | 1.84 b ± 0.28 | 2.07 b ± 0.30 |
BCF5 | 10.53 a ± 1.69 | 8.51 a ± 1.29 | 9.21 a ± 0.96 |
MOT30 | 50.28 a ± 15.09 | 41.57 a ± 16.01 | 37.80 a ± 14.87 |
PMOT30 | 36.91 a ± 11.90 | 31.01 a ± 16.46 | 27.42 a ± 14.28 |
VCL30 | 78.24 a ± 12.57 | 74.79 a ± 14.30 | 69.44 a ± 14.42 |
VSL30 | 59.79 a ± 9.19 | 60.16 a ± 11.18 | 58.00 a ± 13.43 |
VAP30 | 66.33 a ± 10.66 | 65.76 a ± 13.09 | 62.23 a ± 14.10 |
LIN30 | 76.82 a ± 7.29 | 80.75 a ± 7.38 | 83.54 a ± 7.71 |
STR30 | 90.37 a ± 4.59 | 91.67 a ± 3.90 | 93.21 a ± 4.16 |
ALH30 | 2.33 a ± 0.59 | 1.95 a ± 0.60 | 1.83 a ± 0.41 |
BCF30 | 9.76 a ± 1.45 | 8.71 a ± 1.34 | 8.48 a ± 1.67 |
EXTENDER | Botucrio® | Inra-Freeze® | Lac-Edta |
---|---|---|---|
MOT5 | 73.48 a ± 5.71 | 57.33 a ± 18.71 | 46.45 a ± 25.63 |
PMOT5 | 61.70 a ± 6.49 | 42.17 b ± 11.01 | 32.90 b ± 17.72 |
VCL5 | 93.24 a ± 13.10 | 71.38 b ± 7.07 | 87.61 a ± 13.99 |
VSL5 | 66.40 a ± 6.86 | 53.46 b ± 4.30 | 59.26 a,b ± 4.75 |
VAP5 | 72.49 a ± 6.05 | 63.88 a ± 10.02 | 70.70 a ± 7.70 |
LIN5 | 72.45 a ± 13.12 | 75.07 a ± 3.71 | 68.47 a ± 6.61 |
STR5 | 91.60 a ± 4.01 | 90.89 a ± 3.73 | 84.20 b± 5.86 |
ALH5 | 2.76 a ± 0.81 | 2.41 a ± 0.32 | 2.55 a ± 0.38 |
BCF5 | 11.72 a ± 2.26 | 9.99 b ± 0.95 | 9.85 b ± 1.05 |
MOT30 | 59.27 a ± 12.90 | 50.66 a ± 11.84 | 40.92 a ± 15.36 |
PMOT30 | 46.48 a ± 12.29 | 33.65 b ± 6.61 | 30.60 b ± 10.78 |
VCL30 | 85.67 a ± 9.34 | 65.51 b ± 14.02 | 80.55 a ± 8.20 |
VSL30 | 64.54 a ± 6.16 | 51.61 b ± 9.31 | 63.23 a ± 6.37 |
VAP30 | 69.88 a ± 5.78 | 57.73 b ± 12.38 | 71.38 a ± 8.07 |
LIN30 | 75.92 a ± 9.71 | 75.92 a ± 7.03 | 78.63 a ± 5.56 |
STR30 | 92.33 a ± 3.15 | 90.02 a ± 6.04 | 88.75 a ± 4.15 |
ALH30 | 2.59 a ± 0.67 | 2.16 a± 0.59 | 2.23 a ± 0.49 |
BCF30 | 10.96 a ± 1.16 | 9.20 b ± 1.57 | 9.13 b ± 0.86 |
EXTENDER | Botucrio® | Inra-Freeze® | Lac-Edta |
---|---|---|---|
MOT5 | 64.14 a ± 6.79 | 43.59 b ± 11.30 | 38.30 b ± 13.46 |
PMOT5 | 55.69 a ± 11.76 | 34.43 b ± 10.04 | 27.11 b ± 8.16 |
VCL5 | 79.62 a ± 8.86 | 71.90 a ± 16.64 | 82.54 a ± 9.29 |
VSL5 | 66.31 a ± 11.29 | 62.39 a ± 15.22 | 64.27 a ± 9.67 |
VAP5 | 70.94 a ± 10.90 | 66.11 a ± 15.30 | 73.53 a ± 9.39 |
LIN5 | 83.00 a ± 7.83 | 86.61 a ± 2.36 | 77.56 b ± 6.13 |
STR5 | 93.34 a ± 3.90 | 94.16 a ± 2.01 | 87.43 b ± 6.12 |
ALH5 | 1.96 a ± 0.27 | 1.90 a ± 0.39 | 1.94 a ± 0.28 |
BCF5 | 9.26 a ± 1.08 | 8.54 a ± 1.58 | 8.63 a ± 0.83 |
MOT30 | 54.03 a ± 5.96 | 35.86 b ± 17.09 | 29.80 b ± 10.85 |
PMOT30 | 42.03 a ± 11.30 | 23.49 b ± 17.22 | 22.19 b ± 10.01 |
VCL30 | 72.93 a ± 10.70 | 65.76 a ± 10.49 | 79.95 a ± 19.75 |
VSL30 | 61.63 a ± 11.59 | 54.01 a ± 10.44 | 61.79 a ± 13.85 |
VAP30 | 65.95 a ± 11.76 | 57.18 ± 10.09 | 69.81 a ± 16.67 |
LIN30 | 84.35 a ± 8.82 | 82.03 a ± 6.57 | 78.01 a ± 6.63 |
STR30 | 93.43 a ± 4.09 | 94.23 a ± 2.42 | 88.70 a ± 3.55 |
ALH30 | 1.80 a ± 0.40 | 1.85 a ± 0.60 | 2.11 a ± 0.64 |
BCF30 | 8.46 a ± 1.03 | 9.00 a ± 0.79 | 8.44 a ± 2.12 |
Stallion | Number of Frozen Ejaculates | Botucrio® | Inra-Freeze® | Lac EDTA |
---|---|---|---|---|
Stallion A | 3 | 62.67 | 42.83 | 17.77 |
Stallion B | 1 | 65.8 | 46.4 | 39.7 |
Stallion C | 2 | 64.77 | 32.97 | 34.17 |
Stallion D | 2 | 55.6 | 33.3 | 25.6 |
Stallion E | 1 | 39.6 | 36.2 | 30.3 |
Stallion F | 3 | 54.2 | 38.75 | 36.8 |
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Gutiérrez-Cepeda, L.; Crespo, F.; Blazquez, J.C.; Serres, C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals 2023, 13, 382. https://doi.org/10.3390/ani13030382
Gutiérrez-Cepeda L, Crespo F, Blazquez JC, Serres C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals. 2023; 13(3):382. https://doi.org/10.3390/ani13030382
Chicago/Turabian StyleGutiérrez-Cepeda, Luna, Francisco Crespo, Juan Carlos Blazquez, and Consuelo Serres. 2023. "Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation" Animals 13, no. 3: 382. https://doi.org/10.3390/ani13030382
APA StyleGutiérrez-Cepeda, L., Crespo, F., Blazquez, J. C., & Serres, C. (2023). Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals, 13(3), 382. https://doi.org/10.3390/ani13030382