Egg yolk plasma can replace egg yolk in stallion freezing extenders.
Abstract: Hen egg yolk is normally used as a cryoprotective agent in semen freezing extenders, but its use has sanitary and practical disadvantages. Moreover the protection afforded by egg yolk has not yet been completely elucidated. The objective of this study was to compare the egg yolk plasma fraction to whole egg yolk in stallion freezing extender. Plasma contains mainly Low Density Lipoproteins (LDL), which are widely presumed to be the cryoprotective agent in egg yolk. Plasma can be produced on an industrial scale, sterilised by gamma-irradiation and incorporated in a ready-to-use extender (our ultimate objective). Plasma samples were subjected to different doses of gamma-irradiation (3, 5, 10 kGy) without dramatic chemical changes that may affect their cryoprotective properties. Stallion semen was frozen with whole egg yolk as a control and with sterilised egg yolk plasma. A fertility trial was conducted on a total of 70 mares' cycles. Fertility per cycle was 60% after insemination of semen frozen in our control extender containing egg yolk (EY), compared to 69% for the extender containing sterilised egg yolk plasma (EYP) (P > 0.05). Post-thaw motility and membrane integrity of spermatozoa were also analysed. Motility parameters were not significantly different between extenders except for the variable VAP (for EY versus EYP, VAP: 63 μm.s(-1) versus 59 μm.s(-1), a, b: P 0.05). Membrane integrity was better preserved in EY than in EYP but the difference between extenders was small (P < 0.05). Our results demonstrated that sterilised egg yolk plasma has the potential to replace egg yolk in stallion freezing extender. This experiment led to the development of a ready-to-use extender called INRA-Freeze(®) (IMV-Technologies, France).
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication Date: 2010-09-15 PubMed ID: 20833417DOI: 10.1016/j.theriogenology.2010.07.015Google Scholar: Lookup
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- Evaluation Study
- Journal Article
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- Non-U.S. Gov't
Summary
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This study explored the use of egg yolk plasma as a replacement for whole egg yolk in the freeze preservation of stallion semen. Researchers found that egg yolk plasma can potentially provide the same cryoprotective properties as whole egg yolk, thereby leading to the development of a new, ready-to-use preservation solution.
Objective of the Study
- The objective of the study was to evaluate the effectiveness of egg yolk plasma in the freezing and preservation of stallion semen. Egg yolk has been traditionally used as a cryoprotective agent, yet it presents certain sanitary and practical challenges. Egg yolk also contains low density lipoproteins (LDL), which are believed to be the primary component providing cryoprotection. Thus, the researchers aimed to compare the impact of egg yolk plasma (which contains LDL) and whole egg yolk on the preservation of stallion semen.
Methodology
- Plasma samples were subjected to different levels of gamma-irradiation (3, 5, 10 kGy) to sterilize them without affecting their cryoprotective properties.
- Stallion semen was frozen using both traditional whole egg yolk as a control and the sterilized egg yolk plasma for comparison.
- A fertility trial was performed involving 70 mares’ cycles to evaluate the effectiveness of the extenders (a substance used to dilute and preserve semen).
Results
- The use of sterilized egg yolk plasma resulted in a slightly higher fertility per cycle (69%) compared to the control extender containing whole egg yolk (60%). However, this difference wasn’t significant statistically.
- Post-thaw motility and membrane integrity of spermatozoa were also analyzed. Motility parameters were not significantly different between the two groups, except for the VAP (velocity average path) which was slightly lower in the plasma group. However, the difference was very small.
- Membrane integrity was slightly better preserved in the whole egg yolk group than in the plasma group, but the difference was minor.
Conclusion
- The results from the experiment indicated that sterilized egg yolk plasma has potential as a replacement for whole egg yolk in stallion semen freezing extenders, as it provides similar cryoprotection and resulted in comparable fertility rates.
- Based on these findings, the researchers developed a new, ready-to-use extender called INRA-Freeze®, provided by IMV-Technologies, France.
Cite This Article
APA
Pillet E, Duchamp G, Batellier F, Beaumal V, Anton M, Desherces S, Schmitt E, Magistrini M.
(2010).
Egg yolk plasma can replace egg yolk in stallion freezing extenders.
Theriogenology, 75(1), 105-114.
https://doi.org/10.1016/j.theriogenology.2010.07.015 Publication
Researcher Affiliations
- INRA, UMR85 Physiologie de la Reproduction et des Comportements, Nouzilly, France.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents
- Egg Yolk
- Female
- Horses
- Lipoproteins, LDL / physiology
- Male
- Nanoparticles / chemistry
- Oxidative Stress
- Particle Size
- Pregnancy
- Pregnancy Rate
- Semen Analysis
- Semen Preservation / methods
- Semen Preservation / veterinary
- Spermatozoa / physiology
- Spermatozoa / ultrastructure
- Sterilization
Citations
This article has been cited 14 times.- Benko F, Mohammadi-Sangcheshmeh A, Ďuračka M, Lukáč N, Tvrdá E. In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences. PLoS One 2022;17(10):e0276683.
- Cheng Q, Li L, Jiang M, Liu B, Xian Y, Liu S, Liu X, Zhao W, Li F. Extend the Survival of Human Sperm In Vitro in Non-Freezing Conditions: Damage Mechanisms, Preservation Technologies, and Clinical Applications. Cells 2022 Sep 12;11(18).
- Akhtar MF, Ma Q, Li Y, Chai W, Zhang Z, Li L, Wang C. Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology. Animals (Basel) 2022 Sep 2;12(17).
- Mahiddine FY, Kim MJ. Overview on the Antioxidants, Egg Yolk Alternatives, and Mesenchymal Stem Cells and Derivatives Used in Canine Sperm Cryopreservation. Animals (Basel) 2021 Jun 28;11(7).
- Snoeck PPDN, Pessoa THO, Pereira MGS, Bastos ICL, de Melo MIV. Can we use LDL instead of egg yolk in BotuCrio® extender to cryopreserve sperm from the Mangalarga Marchador stallion?. Anim Reprod 2019 Oct 23;16(2):340-347.
- Charbonneau AM, Kinsella JM, Tran SD. 3D Cultures of Salivary Gland Cells in Native or Gelled Egg Yolk Plasma, Combined with Egg White and 3D-Printing of Gelled Egg Yolk Plasma. Materials (Basel) 2019 Oct 24;12(21).
- Rusco G, Di Iorio M, Gibertoni PP, Esposito S, Penserini M, Roncarati A, Cerolini S, Iaffaldano N. Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo. Animals (Basel) 2019 May 31;9(6).
- Hall SE, Negus C, Johinke D, Bathgate R. Adjusting cryodiluent composition for improved post-thaw quality of rabbit spermatozoa. PLoS One 2017;12(4):e0175965.
- Varma VP, Devi L, Venna NK, Murthy ChL, Idris MM, Goel S. Ocular Fluid As a Replacement for Serum in Cell Cryopreservation Media. PLoS One 2015;10(7):e0131291.
- Gutiérrez-Cepeda L, Fernández A, Crespo F, Ramírez MÁ, Gosálvez J, Serres C. The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA. Acta Vet Scand 2012 Dec 5;54(1):72.
- 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 (Basel) 2024 Aug 25;14(17).
- Ivanova A, Simonenko E, Yakovenko S, Spiridonov V. Problems of human spermatozoa cryopreservation: research methods, solutions. Biophys Rev 2023 Oct;15(5):1223-1232.
- Seyedasgari F, Asadi B, Kim E. Seminal plasma modulates post-thaw longevity and motility of frozen sperm in dromedary camel. Anim Biosci 2023 Dec;36(12):1821-1830.
- Nazeri E, Niasari-Naslaji A, Ghasemzadeh-Nava H, Panahi F. The effect of varying concentrations of chicken plasma egg yolk and glycerol on the viability of canine sperm following short and long-term preservation. Iran J Vet Res 2023;24(1):6-13.
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