Alternative Opportunities to Collect Semen and Sperm Cells for Ex Situ In Vitro Gene Conservation in Sheep
Abstract
:1. Introduction
Historical Background
2. Alternative Semen Collection Methods
2.1. Semen Collection by Electroejaculation
2.1.1. Distress and Animal Welfare Issues Regarding Electroejaculation
2.1.2. The Quality of Electroejaculated Ram Spermatozoa
2.2. Sperm Collection after Castration or Postmortem
The Quality of Ram Epididymal Spermatozoa
3. Techniques for Enhancing Post-Thaw Sperm Quality Prior to Freezing
3.1. Treating Sperm Cells with Certain Biological Compounds or Chemicals
3.2. Treating Sperm Cells with Certain Ions
4. Freezing of Electroejaculated and Epididymal Ram Spermatozoa
4.1. Freezing Electroejaculated Spermatozoa
4.2. Freezing Epididymal Spermatozoa
- Straws (fine paillette, 0.25 mL, Tiefenbach, Germany) were cooled from room temperature (20 °C) to 4 °C, at a rate of −0.26 °C/min.
- The equilibration period was shortened to 1 h instead of 2 h.
- Freezing of samples at 4 cm above the liquid nitrogen (LN2) vapor was extended to 30 min as opposed to the normal period of 8–10 min.
- The thawing period was shortened to 17 s at 37 °C instead of 30 s at 37 °C.
5. Conclusions
- Semen concentration (highest reported in the 21st century; 5.2 × 109/mL, compared to that recorded in the 20th century: 2.4 × 109/mL).
- Animal welfare: probe type and voltage used (most of the 20th-century studies used bipolar rectal electrode probes that deliver 30 volts or probe types with ring electrodes, compared to the 21st-century studies, which mainly used three-longitudinal electrode probes, delivering a maximum of 10–15 volts).
- Stimulation duration (used up to 10 min in the 20th-century studies and a maximum of 5 min in the 21st-century studies).
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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S/N | Century | Probe Description | Voltage (V) | Duration (s) | Rest Time (s) | Concentration (106/mL) | References |
---|---|---|---|---|---|---|---|
1 | 20th | Bipolar electrode | 30 | 5 | 5 | 1400 | [61] |
2 | Bipolar electrode | 30 | 5 | 5 | 229 | [22] | |
3 | Bipolar electrode | 30 | 8,5, and 3 | NA | 2500 | [61] | |
4 | Bipolar electrode | 8 | 40 c/s | NA | 1100 | [54] | |
5 | 2 Brass RE 22 cm L × 2.5 cm D | 5.1 | 3 | 7 | 1900 | [62] | |
6 | Ruakura probe | 11 | 10 | 0.02 | 2400 | [63] | |
7 | 21st | 3-LE 22 cm L × 2.5 cm D | NA | 2 | 2 | 5200 | [37] |
8 | 3-LE 23 cm L × 2.5 cm D | 8 | 3 | NA | NA | [55] | |
9 | 3-LE 25 cm L × 3 cm D | 4 | NA | NA | NA | [50] | |
10 | RE 16.5 cm L × 1.7 cm D | 9 | 2–5 | 5 | NA | [64] | |
11 | 3-LE 22 cm L × 2.5 cm D | NA | 2 | 2 | NA | [65] | |
12 | 3-LE 22 cm L × 2.5 cm D | NA | 5 | 5 | NA | [11] | |
13 | 3-LE 35 cm L × 3.2 cm D | 5 | 5 | 5 | 4900 | [38] | |
14 | 3-LE 25 cm L × 2.5 cm D | 9 | 5 | 5 | NA | [66] |
(a) | |||||||||||||||||||||||||||||||||||
Fresh | |||||||||||||||||||||||||||||||||||
Collection Methods | Volume | Concentration (/mL) | Functional Mitochondria | Intact Plasma Membrane | Abnormal Sperm (%) | Total Motility (%) | Progressive Motility (%) | References | |||||||||||||||||||||||||||
EEJ | 1.46 | 1.4 × 109 a | NA | NA | NA | NA | NA | [61] | |||||||||||||||||||||||||||
AV | 1.18 | 1.9 × 109 b | NA | NA | NA | NA | NA | ||||||||||||||||||||||||||||
EEJ | 1.12 | 1.1 × 109 a | NA | NA | 15.3 | 76 a | NA | [22] | |||||||||||||||||||||||||||
AV | 0.71 | 2.7 × 109 b | NA | NA | 16.4 | 84 b | NA | ||||||||||||||||||||||||||||
EEJ | 1.4 a | 2.5 × 109 a | NA | NA | NA | NA | NA | [75] | |||||||||||||||||||||||||||
AV | 1.0 b | 4.3 x109 b | NA | NA | NA | NA | NA | ||||||||||||||||||||||||||||
EEJ | 1.3 | 1.1 × 106 a | NA | NA | 4.4 | NA | NA | [67] | |||||||||||||||||||||||||||
AV | 1.1 | 1.7 × 106 b | NA | NA | 5.7 | NA | NA | ||||||||||||||||||||||||||||
EEJ | 1.1 | 2.34 × 109 a | NA | NA | 4.0 | 77.18 a | NA | [76] | |||||||||||||||||||||||||||
AV | 1.2 | 3.10 × 109 b | NA | NA | 3.9 | 86.7 b | NA | ||||||||||||||||||||||||||||
EEJ | 1.1 | 5.2 × 109 a | NA | NA | 8.1 | 71.9 | NA | [37] | |||||||||||||||||||||||||||
AV | 1.2 | 6.2 × 109 b | NA | NA | 9.1 | 69.8 | NA | ||||||||||||||||||||||||||||
EEJ | NA | NA | NA | NA | NA | 74.3 | 46.6 b | [50] | |||||||||||||||||||||||||||
AV | NA | NA | NA | NA | NA | 77.5 | 59.0 a | ||||||||||||||||||||||||||||
EEJ | 3.99 a | NA | 47.7 a | 71.3 a | NA | 81.7 | 77.5 | [65] | |||||||||||||||||||||||||||
AV | 1.74 b | NA | 16.7 b | 49.5 b | NA | 71.7 | 78.3 | ||||||||||||||||||||||||||||
EEJ | NA | NA | 33.2 | 68.0 a | NA | 67.5 a | 76.0 a | [11] | |||||||||||||||||||||||||||
AV | NA | NA | 27.4 | 47.7 b | NA | 46.7 b | 52.0 b | ||||||||||||||||||||||||||||
EEJ | 1.25 a | 1.7 × 109 a | NA | 55.3 a | NA | 76.04 | 67.52 a | [77] | |||||||||||||||||||||||||||
AV | 0.79 b | 4.9 × 109 b | NA | 70.5 b | NA | 74.79 | 53.67 b | ||||||||||||||||||||||||||||
(b) | |||||||||||||||||||||||||||||||||||
Post-Thaw | |||||||||||||||||||||||||||||||||||
Collection Methods | Viability (%) | TM (%) | PM (%) | LN (%) | LR (%) | Linearity (%) | IPM (%) | IA (%) | DA (%) | FM (%) | References | ||||||||||||||||||||||||
EEJ | 30.4 | 39.6 | NA | 24.2 a | 1.4 a | 60.3 | NA | 26.0 a | 75.4 | NA | [37] | ||||||||||||||||||||||||
AV | 28.5 | 40.3 | NA | 20.7 b | 3.4 b | 54.1 | NA | 21.2 b | 72.6 | NA | |||||||||||||||||||||||||
EEJ | 24.95 | 28.5 | 7.67 | NA | NA | 39.37 | NA | 21.65 | 31.1 a | NA | [77] | ||||||||||||||||||||||||
AV | 25.39 | 29.92 | 8.21 | NA | NA | 40.80 | NA | 23.31 | 43.5 b | NA | |||||||||||||||||||||||||
EEJ | 25.5 | 48.7 b | 27.6 b | NA | NA | 65.3 | NA | NA | 34.2 | NA | [69] | ||||||||||||||||||||||||
AV | 29.3 | 50.1 a | 38.7 a | NA | NA | 68.4 | NA | NA | 33.5 | NA | |||||||||||||||||||||||||
EEJ | NA | 34.8 | 36.2 | NA | NA | NA | 43.3 a | NA | NA | 20.6 a | [11] | ||||||||||||||||||||||||
AV | NA | 36.4 | 35.6 | NA | NA | NA | 41.1 b | NA | NA | 29.1 b |
Pre-Freeze (%) | Post-Thaw (%) | Recovery Rate (%) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
Parameters | AV | EEJ | ESR | AV | EEJ | ESR | AV | EEJ | ESR | References |
TM | 74.2 | 71.9 | 80.3 | 52.5 | 40.3 | 69.8 | 70.8 | 56,1 | 87.0 | [37,40,123] |
77.5 | 76.0 | 79.9 | 59.1 | 28.5 | 62.0 | 76.3 | 37.5 | 77.6 | [50,77] | |
82.5 | 74.3 | 82.8 | 47.5 | 48.7 | 67.2 | 57.6 | 65.5 | 81.2 | [50,111,115] | |
87.6 | 67.5 | 77.5 | 63.2 | 34.8 | 53.3 | 72.1 | 51.6 | 68.8 | [11,82,124] | |
PM | ||||||||||
70.8 | 67.52 | 68.4 | 45.4 | 7.7 | 55.8 | 64.1 | 11.4 | 82.0 | [40,77,124] | |
78.0 | 46.6 | 74.0 | 54.0 | 27.6 | 48.0 | 69.2 | 59.2 | 64.9 | [71,84] | |
59.0 | 76.0 | 57.6 | 38.7 | 36.2 | 41.5 | 65.6 | 47.6 | 72.0 | [11,50] | |
52.0 | NA | 66.4 | 35.6 | NA | 54.3 | 68.5 | NA | 81.8 | [11,50,115] | |
NA | NA | 75.8 | NA | NA | 45.0 | NA | NA | 59.4 | [82,115] | |
IA | ||||||||||
90.4 | NA | 84.3 | 73.1 | NA | 62.7 | 80.9 | NA | 74.4 | [40,111] | |
91.6 | NA | 91.8 | 80.6 | NA | 79.4 | 88.0 | NA | 86.5 | [82,123] | |
NA | NA | 92.4 | NA | NA | 74.6 | NA | NA | 80.7 | [111] | |
IPM | ||||||||||
84.6 | 55.3 | 84.2 | 60.9 | NA | 57.5 | 71.9 | NA | 68.3 | [40,77,111] | |
84.1 | 68.0 | 88.1 | 65.5 | 43.3 | 75.4 | 77.9 | 63.7 | 85.6 | [11,84] | |
66.9 | NA | 85.3 | 51.7 | NA | 61.1 | 77.3 | NA | 71.6 | [82,122,125] | |
47.7 | NA | 88.7 | 41.1 | NA | 53.6 | 86.2 | NA | 60.4 | [11,115,124] | |
NA | NA | 86.4 | NA | NA | 57.2 | NA | NA | 66.2 | [125] |
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Mujitaba, M.A.; Egerszegi, I.; Kútvölgyi, G.; Nagy, S.; Vass, N.; Bodó, S. Alternative Opportunities to Collect Semen and Sperm Cells for Ex Situ In Vitro Gene Conservation in Sheep. Agriculture 2022, 12, 2001. https://doi.org/10.3390/agriculture12122001
Mujitaba MA, Egerszegi I, Kútvölgyi G, Nagy S, Vass N, Bodó S. Alternative Opportunities to Collect Semen and Sperm Cells for Ex Situ In Vitro Gene Conservation in Sheep. Agriculture. 2022; 12(12):2001. https://doi.org/10.3390/agriculture12122001
Chicago/Turabian StyleMujitaba, Malam Abulbashar, István Egerszegi, Gabriella Kútvölgyi, Szabolcs Nagy, Nóra Vass, and Szilárd Bodó. 2022. "Alternative Opportunities to Collect Semen and Sperm Cells for Ex Situ In Vitro Gene Conservation in Sheep" Agriculture 12, no. 12: 2001. https://doi.org/10.3390/agriculture12122001