Effect of egg yolk-based extender and seminal plasma removal on sperm viability of cooled donkey semen

Authors

  • Carolina Natalia Alonso Universidad Nacional de Río Cuarto, Facultad de Agronomía y Veterinaria, Laboratorio de Producción Equina, Río Cuarto, Córdoba, Argentina https://orcid.org/0000-0001-9794-0821
  • Catalina Castañeira Universidad Nacional de Río Cuarto, Facultad de Agronomía y Veterinaria, Laboratorio de Producción Equina, Río Cuarto, Córdoba, Argentina
  • Ana Flores Bragulat Universidad Nacional de Río Cuarto, Facultad de Agronomía y Veterinaria, Laboratorio de Producción Equina, Río Cuarto, Córdoba, Argentina
  • Luis Losinno Universidad Nacional de Río Cuarto, Facultad de Agronomía y Veterinaria, Laboratorio de Producción Equina, Río Cuarto, Córdoba, Argentina

DOI:

https://doi.org/10.11606/issn.1678-4456.bjvras.2021.174301

Keywords:

Donkey, Cooled semen, Semen preservation, Seminal plasma, Egg yolk extender

Abstract

Developing effective cooled semen protocols is essential to increase pregnancy rates and reproductive efficiency in donkeys. This study aimed to evaluate the effect on sperm kinetic parameters and membrane integrity in cooled donkey semen diluted with defined milk proteins extender with 1% or 2% of egg yolk and the removal of seminal plasma. Twenty-four ejaculates from six jackasses were collected. Each ejaculate was divided into four aliquots that were diluted in extender with 1% (EY1) or 2% (EY2) egg yolk. One sample from each group was centrifuged, seminal plasma was removed (CEY1, CEY2 groups, respectively), and the samples were then refrigerated at 5 °C for 24 h. Fresh and cooled semen samples were assessed for sperm motility, morphology, and plasma membrane integrity. Total motility, progressive motility, sperm kinetic parameters, or live sperm cells were not statistically different when semen was cooled with an extender supplemented with 1% or 2% of egg yolk. Seminal plasma removal does not affect total motility or sperm kinetic parameters. However, progressive motility decreased (P<0.05) when semen was extended with 2% of egg yolk and seminal plasma was removed. Membrane integrity was affected (P<0.05) in centrifuged samples. In conclusion, the obtained results suggest that there is no difference in sperm kinetics and membrane integrity when 1% or 2% of egg yolk was added to the Equiplus(R) extender. Also, the removal of seminal plasma by centrifugation did not have any beneficial effect on cooled donkey semen. Further studies are needed to relate these results with in vivo fertility tests with cooled donkey semen.

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References

Ali SS, Marcondes MCG, Bajova H, Dugan LL, Conti B. Metabolic depression and increased reactive oxygen species production by isolated mitochondria at moderately lower temperatures. Journal of Biological Chemistry. 2010;285(42):32522-8. https://doi.org/10.1074/jbc.M110.155432. PMid:20716522.

Alonso C, Castañeira C, Castex CB, Ferrante A, Arraztoa C, Flores A, Gambarotta M, Miragaya M, Losinno L. Effects of extenders and centrifugation on viability of donkey cooled semen. Preliminary Results. Proceedings of the 1st International Symposium on Donkey Science. 2017. Aug 14-17; Shangai China: Dong’e E Jiao Co, Shandong. 196–200.

Alvarez AL, Serres C, Crespo F, Santiago I, Mateos EG-CC. Natural service and artificial insemination with cooled semen of Zamorano-Leones donkey. Proceedings of the 15th International Congress Animal Reproduction; 2004 Aug 8-12; Porto Seguro, 2004.

Aurich C. Factors affecting the plasma membrane function of cooled-stored stallion spermatozoa. Anim. Reprod. Sci. 2005;89(1-4 Spec Issue):65-75.

Batellier F, Duchamp G, Vidament M, Arnaud G, Palmer E, Magistrini M. Delayed insemination is successful with a new extender for storing fresh equine semen at 15 °C under aerobic conditions. Theriogenology. 1998;50(2):229-36. https://doi.org/10.1016/S0093-691X(98)00130-7. PMid:10734490.

Batellier F, Magistrini M, Fauquant J, Palmer E. Effect of milk fractions on survival of equine spermatozoa. Theriogenology. 1997;48(3):391-410. https://doi.org/10.1016/S0093-691X(97)00250-1. PMid:16728137.

Batellier F, Vidament M, Fauquant J, Duchamp G, Arnaud G, Yvon JM, Magistrini M. Advances in cooled semen technology. Anim Reprod Sci. 2001;68(3–4):181-90. https://doi.org/10.1016/S0378-4320(01)00155-5. PMid:11744263.

Bergeron A, Crête M-H, Brindle Y, Manjunath P. Low-density lipoprotein fraction from Hen’s egg yolk decreases the binding of the major proteins of bovine seminal plasma to sperm and prevents lipid efflux from the Sperm Membrane 1. Biol Reprod. 2004;70(3):708-17. https://doi.org/10.1095/biolreprod.103.022996. PMid:14613896.

Bergeron A, Manjunath P. New insights towards understanding the mechanisms of sperm protection by egg yolk and milk. Mol Reprod Dev. 2006;73(10):1338-44. https://doi.org/10.1002/mrd.20565. PMid:16868925.

Bergeron A, Villemure M, Lazure C, Manjunath P. Isolation and characterization of the major proteins of ram seminal plasma. Mol Reprod Dev. 2005;71(4):461-70. https://doi.org/10.1002/mrd.20310. PMid:15892046.

Calvete JJ, Nessau S, Mann K, Sanz L, Sieme H, Klug E, Töpfer-Petersen E. Isolation and biochemical characterization of stallion seminal-plasma proteins. Reprod Domest Anim. 1994;29(5):411-26. https://doi.org/10.1111/j.1439-0531.1994.tb00588.x.

Camillo F, Rota A, Biagini L, Tesi M, Fanelli D, Panzani D. The current situation and trend of donkey industry in Europe. J Equine Vet Sci. 2018;65:44-9. https://doi.org/10.1016/j.jevs.2017.11.008.

Carvalho LE, Silva Filho JM, Palhares MS, Sales ALR, Gonczarowska AT, Oliveira HN, Resende MC, Neves MG, Madison RJ. Physical characteristics and fertility of fractionated donkey semen cooled at 5 °C. Arq Bras Med Vet Zootec. 2017;69(1):29-38. https://doi.org/10.1590/1678-4162-7976.

Choong C, Wales R. The Effect of Cold Shock on Spermatozoa. Aust J Biol Sci. 1962;15(3):543-51. https://doi.org/10.1071/BI9620543.

Cochran JD, Amann RP, Froman DP, Pickett BW. Effects of centrifugation, glycerol level, cooling to 5 degrees C, freezing rate and thawing rate on the post-thaw motility of equine sperm. Theriogenology. 1984;22(1):25-38. https://doi.org/10.1016/0093-691X(84)90470-9 PMid:16725933.

Cottorello ACP, Amancio RC, Henry M, Borges I. Effect of storage temperature and extenders on “in vitro” activity of donkey spermatozoa. Theriogenology. 2002;88:325-8.

De Leeuw FE, De Leeuw AM, Den Daas JHG, Colenbrander B, Verkleij AJ. Effects of various cryoprotective agents and membrane-stabilizing compounds on bull sperm membrane integrity after cooling and freezing. Cryobiology. 1993;30(1):32-44. https://doi.org/10.1006/cryo.1993.1005. PMid:8440128.

Di Palma T, Cecchini S, MacChia G, Pasolini MP, Cocchia N, Boni R. Kinematic, bioenergetic and oxidative evaluations of donkey sperm preserved at +4°C. Zygote. 2020;28(4):300-7. https://doi.org/10.1017/S096719942000012X. PMid:32285764.

Dini P, Troch L, Lemahieu I, Deblende P, Daels P. Validation of a portable device (iSperm) for the assessment of stallion sperm motility and concentration. Reprod Domest Anim. 2019;54(8):1113-20. https://doi.org/10.1111/rda.13487. PMid:31177582.

Jasko DJ, Hathaway JA, Schaltenbrand VL, Simper WD, Squires EL. Effect of seminal plasma and egg yolk on motion characteristics of cooled stallion spermatozoa. Theriogenology. 1992;37(6):1241-52. https://doi.org/10.1016/0093-691X(92)90180-Y.

Leboeuf B, Guillouet P, Batellier F, Bernelas D, Bonné JL, Forgerit Y, Renaud G, Magistrini M. Effect of native phosphocaseinate on the in vitro preservation of fresh semen. Theriogenology. 2003;60(5):867-77. https://doi.org/10.1016/S0093-691X(03)00095-5. PMid:12935864.

Martin ICA. Diluents for the preservation of ram spermatozoa 1. Aust J Biol Sci. 1966;19(4):645-53. PMid:5970061.

Mello SLV, Henry M, Souza MC, Oliveira SMP. Effect of split ejaculation and seminal extenders on longevity of donkey semen preserved at 5°C. Arq Bras Med Vet Zootec. 2000;52(4):372-8. https://doi.org/10.1590/S0102-09352000000400015.

Miró J, Taberner E, Rivera M, Peña A, Medrano A, Rigau T, Peñalba A. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen. Theriogenology. 2009;72(8):1017-22. https://doi.org/10.1016/j.theriogenology.2009.06.012. PMid:19747718.

Newcombe J, Cuervo-Arango J. The effect of time of insemination with fresh cooled transported semen and natural mating relative to ovulation on pregnancy and embryo loss rates in the mare. Reprod Domest Anim. 2011;46(4):678-81. https://doi.org/10.1111/j.1439-0531.2010.01728.x. PMid:21114795.

Pagl R, Aurich JE, Müller-Schlösser F, Kankofer M, Aurich C. Comparison of an extender containing defined milk protein fractions with a skim milk-based extender for storage of equine semen at 5°C. Theriogenology. 2006;66(5):1115-22. https://doi.org/10.1016/j.theriogenology.2006.03.006. PMid:16620943.

Phillips PH, Lardy HA. A yolk-buffer pabulum for the preservation of bull semen. J Dairy Sci. 1940;23(5):399-404. https://doi.org/10.3168/jds.S0022-0302(40)95541-2.

R Core Team. (2018) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna. https://www.R-project.org.

Rota A, Bastianacci V, Magelli C, Panzani D, Camillo F. Evaluation of plasma membrane integrity of donkey spermatozoa. Reprod Domest Anim. 2010;45(2):228-32. https://doi.org/10.1111/j.1439-0531.2008.01241.x. PMid:19144042.

Rota A, Furzi C, Panzani D, Camillo F. Studies on motility and fertility of cooled stallion spermatozoa. Reprod Domest Anim. 2004;39(2):103-9. https://doi.org/10.1111/j.1439-0531.2004.00488.x. PMid:15065992.

Rota A, Magelli C, Panzani D, Camillo F. Effect of extender, centrifugation and removal of seminal plasma on cooled-preserved Amiata donkey spermatozoa. Theriogenology. 2008;69(2):176-85. https://doi.org/10.1016/j.theriogenology.2007.09.003. PMid:17945340.

Rota A, Perulli G, Sabatini C, Panzani D, Camillo F. Cooled semen artificial insemination in donkeys: field results. Reprod Domest Anim. 2017;52:127.

Serres C, Rodriguez A, Alvarez AL, Santiago I, Gabriel J, Gómez-Cuétara C, Mateos E. Effect of centrifugation and temperature on the motility and plasma membrane integrity of Zamorano-Leonés donkey semen. Theriogenology. 2002;58(2–4):329-32.

Talluri TR, Mal G, Ravi SK. Biochemical components of seminal plasma and their correlation to the fresh seminal characteristics in Marwari stallions and Poitou jacks. Vet World. 2017;10(2):214-20. https://doi.org/10.14202/vetworld.2017.214-220. PMid:28344405.

Varner DD, Blanchard TL, Meyers PJ, Meyers SA. Fertilizing capacity of equine spermatozoa stored for 24 hours at 5 or 20°C. Theriogenology. 1989;32(4):515-25. https://doi.org/10.1016/0093-691X(89)90273-2. PMid:16726699.

Villemure M, Lazure C, Manjunath P. Isolation and characterization of gelatin-binding proteins from goat seminal plasma. Reprod Biol Endocrinol. 2003;1(1):1-10. https://doi.org/10.1186/1477-7827-1-39. PMid:12737634.

Vyvial M, Horáčková E, Sedlinská M, Jánová E, Krisová Š, Mráčková M. Determination of selected components in seminal plasma of donkey stallions and their correlation to semen quality parameters Seminal plasma is a complex combination of carbohydrates, proteins, minerals, vitamins and enzymes. It is a fluid that is mixe. Acta Vet Brno. 2019;88(4):377-84. https://doi.org/10.2754/avb201988040377.

Wu S, Shao Y, Zhang H, Wang S, Ji C, Zeng S, Ye H, Yu J. The effects of egg yolk concentration and particle size on donkey semen preservation. J Equine Vet Sci. 2018;65:19-24. https://doi.org/10.1016/j.jevs.2018.03.002.

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Published

2021-04-23

How to Cite

Effect of egg yolk-based extender and seminal plasma removal on sperm viability of cooled donkey semen. (2021). Brazilian Journal of Veterinary Research and Animal Science, 58, e174301. https://doi.org/10.11606/issn.1678-4456.bjvras.2021.174301