Replacing egg yolk with soybean lecithin in the cryopreservation of stallion semen.
Abstract: The objective of this study was to determine whether replacing the egg yolk with soybean lecithin in the Botu-Crio® cryodiluent would maintain the fertility of cryopreserved stallion sperm. Two experiments were performed to evaluate cell freezability. In experiment 1, sperm from 15 stallions were frozen in Botu-Crio® (BC) or Botu-Crio® which contained 45g/L soybean lecithin (BCLS45) in place of the egg yolk. In experiment 2, we compared different concentrations of soybean lecithin: 0, 10.0, 12.5, 15.0, 17.5 and 20.0g/L (BC, BCLS10, BCLS12.5, BCLS17.5 and BCLS20, respectively). In experiment 1, sperm frozen in BC and BCLS45 exhibited similar (P>0.05) percentages of total motile sperm (61% and 61%, respectively); progressively motile sperm (27% and 27%, respectively) and sperm with intact plasma membranes (IMP; 53% and 57%, respectively). Similarly, sperm frozen in BC or BC containing any concentration of soybean lecithin maintained similar (P>0.05) percentages of total motile sperm (61-68%) and progressively motile sperm (27-31%). In the first fertility trial, we used cryopreserved semen from a single stallion was inseminated into mares. The semen from the sperm that were frozen in BC diluent resulted in a higher fertility rate (66%, 16/24) compared to the sperm that were frozen in BCLS45 diluent (17%, 5/29; P<0.01). Similarly, in a second fertility trial, the mares that were inseminated with the sperm that were frozen in BC diluent exhibited a higher fertility rate (66%, 16/24) compared to the mares that were inseminated with the sperm that were frozen in BCLS20 (40%, 10/25; P<0.05). Finally, in a third trial, the sperm that were frozen in BC resulted in a higher fertility rate in mares (75%, 18/24) compared to the sperm that were frozen in BCLS10 (41%, 10/24; P<0.05). Although replacing the egg yolk in the BC cryodiluent with soybean lecithin provided similar laboratory results for stallion sperm, after cryopreservation, the sperm that was frozen with soybean lecithin in the diluent correlated with lower fertility rates. Based on these results, we concluded that the use of BCLS can be used as an alternative diluent for cryopreserving stallion sperm. However, the resulting reduced fertility rate is a matter of concern. Further studies are necessary to clarify the reasons for this decrease in fertility and to determine the optimal lecithin concentration for diluents to freeze stallion sperm.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication Date: 2011-10-15 PubMed ID: 22112939DOI: 10.1016/j.anireprosci.2011.10.006Google Scholar: Lookup
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Summary
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The research study investigates the viability of replacing egg yolk with soybean lecithin in a specific cryopreservation solution known as Botu-Crio® when preserving stallion sperm. While lab results showed similar performance levels between the methods, actual fertility rates were lower when using soybean lecithin, necessitating further research.
Experiments and Methodology
- The research was divided into two main experiments. In the first experiment, sperm from 15 stallions were frozen in Botu-Crio® or Botu-Crio® that contained 45g/L soybean lecithin instead of egg yolk.
- In the second experiment, different concentrations of soybean lecithin were compared to find the optimal concentration for sperm preservation.
- Three fertility trials were carried out using the sperm frozen in different concentrations of the diluents, testing the effectiveness of the substitutes by inseminating mares. These fertility rates were then compared.
Results
- In the first experiment, similar percentages (61%) of total motile sperm were found in samples frozen in either diluent. Similar results were also seen for progressively motile sperm (27%) and sperm with intact plasma membranes (53-57%).
- The second experiment suggested that any concentration of soybean lecithin in the Botu-Crio® diluent yielded similar percentages of total motile sperm (61-68%) and progressively motile sperm (27-31%).
- However, the fertility trials found markedly lower fertility rates from sperm frozen with the soybean lecithin diluents. The highest fertility rate achieved with a lecithin diluent was 41%, when using a concentration of 10g/L, while the traditional egg yolk diluent achieved fertility rates of 66% and 75% in two separate trials.
Conclusions
- Although lab tests showed similar performance between the soybean lecithin and egg yolk diluents, the actual fertility rates were considerably lower when using the former. This difference suggests that, while lecithin can serve as an alternative for egg yolk in diluents, its impact on fertility rates is a concern and indicates the need for further investigation.
- Future research should aim to explore the reasons behind the reduced fertility rates and determine the optimal lecithin concentration for cryopreserving stallion sperm.
Cite This Article
APA
Papa FO, Felício GB, Melo-Oña CM, Alvarenga MA, De Vita B, Trinque C, Puoli-Filho JN, Dell'Aqua JA.
(2011).
Replacing egg yolk with soybean lecithin in the cryopreservation of stallion semen.
Anim Reprod Sci, 129(1-2), 73-77.
https://doi.org/10.1016/j.anireprosci.2011.10.006 Publication
Researcher Affiliations
- Department of Animal Reproduction and Veterinary Radiology, São Paulo State University, Botucatu, SP 18610-970, Brazil. papa@fmvz.unesp.br
MeSH Terms
- Animals
- Cell Membrane / physiology
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents
- Female
- Fertility
- Horses
- Insemination, Artificial / veterinary
- Lecithins
- Male
- Pregnancy
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa
Citations
This article has been cited 7 times.- 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).
- Hermansson U, Johannisson A, Axnér E. Cryopreservation of dog semen in a Tris extender with two different 1% soybean preparations compared with a Tris egg yolk extender. Vet Med Sci 2021 May;7(3):812-819.
- Zhao JQ, Xiao GL, Zhu WL, Fang D, Li N, Han CM, Gao QH. Ram semen preserved at 0°C with soybean lecithin Tris-based extender substituted for egg yolk. Anim Biosci 2021 Feb;34(2):192-197.
- 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).
- Burch FC, Nichi M, Mendes CM, Assumpção MEOD, Duarte JMB, Del Rio do Valle R. Cryopreservation of spermatozoa from black-and-gold howler monkeys (Alouatta caraya) using egg yolk-based or soy lecithin-based extenders. Primates 2024 Jul;65(4):341-353.
- Al-Kass Z, Morrell JM. Freezing Stallion Semen-What Do We Need to Focus on for the Future?. Vet Sci 2024 Feb 2;11(2).
- Shabani S, Mehri M, Shirmohammad F, Sharafi M. Enhancement of sperm quality and fertility-related parameters in Hubbard grandparent rooster fed diets supplemented with soybean lecithin and vitamin E. Poult Sci 2022 Mar;101(3):101635.
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