Effect of short-term conservation temperature, with or without centrifugation, on the survival and motility of Catalonian donkey spermatozoa
Abstract
Aim of study: To analyze the effect of three short-term storage temperatures with or without removing seminal plasma on the survival and motility of donkey sperm and the response to refrigeration and centrifugation of the different spermatozoa subpopulations.
Area of study: North-eastern Spain (Catalonia).
Material and methods: Semen from seven Catalonian jackasses was diluted with a skimmed milk-based (Kenney) extender and different treatments were obtained: FRESH semen, FRESH semen immediately centrifuged to remove the seminal plasma before resuspension in Kenney extender (FRESH+CENTRIFUGATION), FRESH semen stored at 5/15/20ºC for 2 h (STORAGE 5/15/20ºC), and STORAGE 5/15/20ºC semen then centrifuged (STORAGE 5/15/20ºC+CENTRIFUGATION). Survival was examined using eosin-nigrosin stained smears. Motion was assessed by means of a computer-assisted sperm analyzer (CASA).
Main results: The spermatozoa of the STORAGE 5ºC and 20ºC showed an overall motility similar to that seen in FRESH samples. However, the STORAGE 15ºC led to an important motility reduction. No differences were seen between the FRESH and STORAGE 5/15/20ºC with respect to progressive motility. However, STORAGE 5/15/20ºC+CENTRIFUGATION all reduced total motility, and STORAGE 15ºC+CENTRIFUGATION led to reduced survival. The sperm motile subpopulations structure of donkey semen was maintained after STORAGE 5/15/20ºC+CENTRIFUGATION, although STORAGE 15ºC+CENTRIFUGATION led to important changes. STORAGE 5/20ºC+CENTRIFUGATION, in contrast, only induced slight changes. STORAGE 20ºC+CENTRIFUGATION was associated with no change in the percentage of sperm cells belonging to each Subpopulation compared to FRESH sperm.
Research highlights 2 h of storage at 20ºC followed by centrifugation is suitable for the short-term storage of donkey semen.
Downloads
References
Abaigar T, Holt W, Harrison R, Del Barrio G, 1999. Sperm subpopulation in boar (Sus scrofa) and gazelle (Gazella dama mhorr) semen as revealed by pattern analysis of computer-assisted motility assessments. Biol Reprod 60: 32-41. https://doi.org/10.1095/biolreprod60.1.32
Amann RP, Pickett BW, 1987. Principle of cryopreservation and a review of cryopreservation of stallion spermatozoa. J Equine Vet Sci 7: 145-173. https://doi.org/10.1016/S0737-0806(87)80025-4
Aranguren-Méndez JA, Jordana J, Gomez M, 2001. Genetic diversity in Spanish donkey breeds using microsatellite DNA markers. Genet Sel Evol 33: 433-442. https://doi.org/10.1186/1297-9686-33-4-433
Aurich C, 2005. Factors affecting the plasma membrane function of cooled-stored stallion spermatozoa. Anim Repro Sci 89: 65-75. https://doi.org/10.1016/j.anireprosci.2005.06.025
Backman T, Bruemmer JE, Graham JK, Squires EL, 2004. Pregnancy rates of mares inseminated with semen cooled for 18 h and then frozen. J Anim Sci 82: 690-694. https://doi.org/10.2527/2004.823690x
Bamba K, 1988. Evaluation of acrosomal integrity of boar spermatozoa by bright field microscopy using an eosin-nigrosin stain. Theriogenology 29: 1245-1251. https://doi.org/10.1016/0093-691X(88)90004-0
Cochran JD, Amann RP, Froman DP, Pickett BW, 1984. Effects of centrifugation, glycerol level, cooling to 5°C, freezing rate and thawing rate on the post-thaw motility of equine sperm. Theriogenology 22: 25-35. https://doi.org/10.1016/0093-691X(84)90470-9
Cottorello ACP, Amancio RC, Henry M, Borges I, 2002. Effect of storage temperature and extenders on "in vitro" activity of donkey spermatozoa. Theriogenology 58: 325-328. https://doi.org/10.1016/S0093-691X(02)00753-7
Crockett EC, Graham JK, Bruemmer JE, Squires EL, 2001. Effect of cooling equine spermatozoa before freezing on post-thaw motility: preliminary results. Theriogenology 55: 793-803. https://doi.org/10.1016/S0093-691X(01)00444-7
Eddy EM, O'Brien DA, 1994. The spermatozoon. In: The physiology of reproduction; Knobil E, Neill JD (Eds.), Raven Press, NY. pp: 29-77.
Flores E, Taberner E, Rivera MM, Peña A, Rigau T, Miró J, Rodríguez-Gil JE, 2008. Effects of freezing/thawing on motile sperm subpopulations of boar and donkey ejaculates. Theriogenology 70: 936-945. https://doi.org/10.1016/j.theriogenology.2008.05.056
Francl AT, Amann RP, Squires EL, Pickett BW, 1987. Motility and fertility of equine spermatozoa in a milk extender after 12 or 24 hours at 20ºC. Theriogenology 27: 517-525. https://doi.org/10.1016/0093-691X(87)90239-1
Martinez-Pastor F, Garcia-Macias V, Alvarez M, Herraez P, Anel L, de Paz P, 2005. Sperm subpopulations in Iberian red deer epididymal sperm and their changes through the cryopreservation process. Biol Reprod 72: 316-327. https://doi.org/10.1095/biolreprod.104.032730
Mátas C, Decuadro G, Martínez-Miró S, Gadea J, 2007. Evaluation of a cushioned method for centrifugation and processing for freezing boar semen. Theriogenology 67: 1087-1091. https://doi.org/10.1016/j.theriogenology.2006.11.010
Mello SLV, Henry M, Souza MC, Oliveira SMP, 2000. Effect of split ejaculation and seminal extenders on longevity of donkey semen preserved at 5ºC. Arq Bras Med Vet Zootec 52: 372-378. https://doi.org/10.1590/S0102-09352000000400015
Miró J, Lobo V, Quintero-Moreno A, Medrano A, Peña A, Rigau T, 2005. Sperm motility patterns and metabolism in Catalonian donkey semen. Theriogenology 63: 1706-1716. https://doi.org/10.1016/j.theriogenology.2004.07.022
Miró J, Taberner E, Rivera M, Peña A, Medrano A, Rigau T, Peñalba A, 2009. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen. Theriogenology 72: 1017-1022. https://doi.org/10.1016/j.theriogenology.2009.06.012
Miró J, Papas M, 2018. Post-Artificial insemination endometrial inflammation and its control in donkeys. J Equin Vet Sci 65: 38-43. https://doi.org/10.1016/j.jevs.2017.11.007
Moran DM, Jasko DJ, Squires EL, Amann RP, 1992. Determination of temperature and cooling rate which induce cold shock in stallion spermatozoa. Theriogenology 38: 999-1012. https://doi.org/10.1016/0093-691X(92)90114-7
Papas M, Arroyo l, Bassols A, Catalán J, Bonilla S, Gacem S, Yeste M, Miró J, 2019. Activities of antioxidant seminal plasma enzymes (SOD, CAT, GPX and GSR) are higher in jackasses than in stallions and are correlated with sperm motility in jackasses. Theriogenology 140: 180-187. https://doi.org/10.1016/j.theriogenology.2019.08.032
Province CA, Squires EL, Pickett BW, Amann RP, 1985. Cooling rates, storage temperatures and fertility of extended equine spermatozoa. Theriogenology 23: 925-934. https://doi.org/10.1016/0093-691X(85)90010-X
Pruitt JA, Arns MJ, Pool KC, 1993. Seminal plasma influences recovery of equine spermatozoa following in vitro culture (37ºC) and cold-storage (5ºC). Theriogenology 39: 291. https://doi.org/10.1016/0093-691X(93)90146-V
Quintero-Moreno A, Miró J, Rigau T, Rodríguez-Gil JE, 2003. Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology 58: 1973-90. https://doi.org/10.1016/S0093-691X(02)01297-9
Rota A, Magelli C, Panzani D, Camillo F, 2008. Effect of extender, centrifugation and removal of seminal plasma on cooled-preserved Amiata donkey spermatozoa. Theriogenology 69: 176-185. https://doi.org/10.1016/j.theriogenology.2007.09.003
Rigau T, Farré M, Ballester J, Mogas T, Peña A, Rodríguez-Gil JE, 2001. Effects of glucose and fructose on motility patterns of dog spermatozoa from fresh ejaculates. Theriogenology 56: 801-815. https://doi.org/10.1016/S0093-691X(01)00609-4
SAS Inst., 2000. SAS/STAC Software. SAS Inst. Inc., Cary, NC, USA.
Serres C, Rodriguez A, Alvarez AL, Santiago I, Gabriel J, Gómez-Cuétara C, Mateos E, 2002. Effect of centrifugation and temperature on the motility and plasma membrane integrity of Zamorano-Leonés donkey semen. Theriogenology 58: 329-332. https://doi.org/10.1016/S0093-691X(02)00876-2
Squires E, Amann RP, McKinnon AO, Pickett BW, 1988. Fertility of equine spermatozoa cooled to 5 or 20ºC. Proc 11th Int Congr on Anim Reprod, Dublin, Ireland, Vol 3, pp: 297-299.
Varner DD, Blanchard TL, Love CL, Garcia MC, Kenney RM, 1987. Effects of semen fractionation and dilution on equine spermatozoal motility parameters. Theriogenology 28: 709-723. https://doi.org/10.1016/0093-691X(87)90288-3
Varner DD, Blanchard TL, Meyers PJ, Meyers SA, 1989. Fertilizing capacity of equine spermatozoa stored for 24 hours at 5 and 20ºC. Theriogenology 32: 515-525. https://doi.org/10.1016/0093-691X(89)90273-2
Vidament M, Ecot P, Noue P, Bourgeois C, Magistrini M, Palmer E, 2000. Centrifugation and addition of glycerol at 22ºC instead of 4ºC improve post-thaw motility and fertility of stallion spermatozoa. Theriogenology 54:907-919. https://doi.org/10.1016/S0093-691X(00)00401-5
© CSIC. Manuscripts published in both the printed and online versions of this Journal are the property of Consejo Superior de Investigaciones Científicas, and quoting this source is a requirement for any partial or full reproduction.
All contents of this electronic edition, except where otherwise noted, are distributed under a “Creative Commons Attribution 4.0 International” (CC BY 4.0) License. You may read here the basic information and the legal text of the license. The indication of the CC BY 4.0 License must be expressly stated in this way when necessary.
Self-archiving in repositories, personal webpages or similar, of any version other than the published by the Editor, is not allowed.
