Short communication. Stallion sperm quality after combined ejaculate fractionation and colloidal centrifugation
Abstract
This study investigated the possible additive benefit of ejaculate fractionation and colloidal centrifugation on stallion sperm quality. Using an open-end artificial vagina, the sperm-rich fraction (FRAC-1) was separated from the rest of the ejaculate (FRAC-2) and a third sperm sample representing the combined ejaculate was reconstituted post-ejaculation (RAW). Each semen sample was processed for colloidal centrifugation. The percentage of abnormal spermatozoa was 17.8 ± 7.0% in RAW and 14.6 ± 9.5% in FRAC-1 but decreased to 11.4 ± 4.7% and 9.6 ± 6.9% respectively, after colloidal centrifugation. A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation. The rate of increase in sperm DNA fragmentation during the first 6 h of incubation at 37 ºC was 1.8 ± 0.9% per hour in RAW semen and 2.0 ± 2.0% per hour in FRAC-1 but this significantly decreased to 1.3 ± 1.4% and 0.9 ± 0.8% respectively after colloidal centrifugation. While stallion seminal characteristics can be improved using colloidal centrifugation, further enhancement is possible if the ejaculate is initially fractionated.
Downloads
References
Ackay E, Reilas T, Andersson M, Katila T, 2006. Effect of seminal plasma fractions on sperm survival after cooled storage. J Vet Med A 53: 481-485. http://dx.doi.org/10.1111/j.1439-0442.2006.00882.x
Colleoni S, Lagutina I, Rodríguez-Martínez H, Lazzari G, Galli C, Morrell JM, 2011. New techniques for selecting spermatozoa for equine assisted reproduction. J Equine Vet Sci 31: 536-541. http://dx.doi.org/10.1016/j.jevs.2011.03.009
Cortés-Gutiérrez EI, Crespo F, Gosálbez A, Dávila-Rodríguez MI, López-Fernández C, Gosálvez J, 2008. DNA fragmentation in frozen sperm of Equus asinus: Zamorano-Leonés, a breed at risk of extinction. Theriogenology 69: 1022-1032. http://dx.doi.org/10.1016/j.theriogenology.2008.02.002
Crespo F, Gutiérrez-Cepeda L, Gosalvez J, Serres C, Johnston S, 2013. Colloidal centrifugation of stallion semen results in a reduced rate of sperm DNA fragmentation. Reprod Domest Anim 48: e23-e25. http://dx.doi.org/10.1111/j.1439-0531.2012.02140.x
Edmond AJ, Brinsko SP, Love CC, Blanchard TL, Teague SR, Varner DD, 2012. Effect of centrifugal fractionation protocols on quality and recovery rate of equine sperm. Theriogenology 77: 959-966. http://dx.doi.org/10.1016/j.theriogenology.2011.10.002
Gutiérrez-Cepeda L, Fernández A, Crespo F, Gósalvez J, Serres C, 2011. Simple and economic colloidal centrifugation protocols may be incorporated to the clinical equine sperm processing procedure. Anim Reprod Sci 124: 85-89. http://dx.doi.org/10.1016/j.anireprosci.2011.02.001
Gutiérrez-Cepeda L, Fernández A, Crespo F, Ramírez MA, Gosálvez J, Serres C, 2012. 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 54: 72. http://dx.doi.org/10.1186/1751-0147-54-72
Johannisson A, Morrell JM, Thorén J, Jonsson M, Dalin AM, Rodríguez-Martínez H, 2009. Colloidal centrifugation with Androcoll-ETM prolongs stallion sperm motility, viability and chromatin integrity. Anim Reprod Sci 116: 119-128. http://dx.doi.org/10.1016/j.anireprosci.2009.01.008
Kareskoski M, Katila T, 2008. Components of stallion seminal plasma and the effects of seminal plasma on sperm longevity. Anim Reprod Sci 107: 249-256. http://dx.doi.org/10.1016/j.anireprosci.2008.04.013
Kareskoski AM, Reilas T, Andersson M, Katila T, 2006. Motility and plasma membrane integrity of spermatozoa in fractionated stallion ejaculates after storage. Reprod Dom Anim 41: 33-38. http://dx.doi.org/10.1111/j.1439-0531.2006.00647.x
Kareskoski AM, Reilas T, Sankari S, Andersson M, Güvenc K, Katila T, 2010. Alkaline and acid phosphatase, β-glucuronidase and electrolyte levels in fractionated stallion ejaculates. Reprod Domest Anim 45: e369-e374. http://dx.doi.org/10.1111/j.1439-0531.2009.01579.x
Kareskoski AM, Rivera del Alamo MM, Güvenc K, Reilas T, Calvete JJ, Rodríguez-Martínez H, Andersson M, Katila T, 2011a. Protein composition of seminal plasma in fractionated stallion ejaculates. Reprod Domest Anim 46: e79-e84. http://dx.doi.org/10.1111/j.1439-0531.2010.01641.x
Kareskoski M, Sankari S, Johannisson A, Kindahl H, Andersson M, Katila T, 2011b. The Association of the presence of seminal plasma and its components with sperm longevity in fractionated stallion ejaculates. Reprod Domest Anim 46: 1073-1081. http://dx.doi.org/10.1111/j.1439-0531.2011.01789.x
Kumar D, Kalthur D, Mascarenhas D, Kumar P, Adiga SK, 2011. Ejaculate fraction of asthenozoospermic and teratozoospermic patients have differences in the sperm DNA integrity. Andrology 43: 416-421. http://dx.doi.org/10.1111/j.1439-0272.2010.01105.x
Lindahl J, Dalin AM, Stuhtmann G, Morrell JM, 2012. Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96 h at 6ºC prior to artificial insemination. Acta Vet Scand 54: 40-44. http://dx.doi.org/10.1186/1751-0147-54-40
Loomis PR, 2006. Advanced methods for handling and preparation of stallion semen. Vet Clin North Am Equine Pract 22: 663-676. http://dx.doi.org/10.1016/j.cveq.2006.07.002
López-Fernández C, Crespo F, Arroyo F, Fernández JL, Arana P, Johnston SD, Gosálvez J, 2007. Dynamics of sperm DNA fragmentation in domestic animals II. The stallion. Theriogenology 68: 1240-1250. http://dx.doi.org/10.1016/j.theriogenology.2007.08.029
Macías-García B, Morrell JM, Ortega-Ferrusola C, González-Fernández L, Tapia JA, Rodríguez-Martínez H, Peña FJ, 2009. Centrifugation on a single layer of colloid selects improved quality spermatozoa from frozen-thawed stallion semen. Anim Reprod Sci 114: 193-202. http://dx.doi.org/10.1016/j.anireprosci.2008.08.025
Magistrini M, Lindeberg H, Koskinen E, Beau P, Seguin F, 2000. Biophysical and 1H magnetic resonance spectroscopy characteristics of fractionated stallion ejaculates. J Reprod Fertil Suppl 56: 101-110.
Mari G, Castagnetti C, Rizzato G, Mislei B, Iacono E, Merlo B, 2011. Density gradient centrifugation of sperm from a subfertile stallion and effect of seminal plasma addition on fertility. Anim Reprod Sci 126: 96 -100. http://dx.doi.org/10.1016/j.anireprosci.2011.04.020
Morrell JM, 2011. Biomimetics in action: Practical applications of single layer centrifugation for equine breeding. J Veterinar Sci Technol 2: 107.
Morrell JM, 2012. Stallion sperm selection: Past, present and future trends. J Equine Vet Sci 32: 436-440. http://dx.doi.org/10.1016/j.jevs.2012.05.069
Morrell JM, Rodríguez-Martínez H, 2009. Biomimetic techniques for improving sperm quality in animal breeding: a review. Open Androl J 1: 1-9.
Morrell JM, Wallgren M, 2011. Removal of bacteria from boar ejaculates by single layer centrifugation can reduce the use of antibiotics in semen extenders. Anim Reprod Sci 123: 64-69. http://dx.doi.org/10.1016/j.anireprosci.2010.11.005
Morrell JM, Johannisson A, Dalin AM, Rodríguez-Martínez H, 2009a. Morphology and chromatin integrity of stallion spermatozoa prepared by density gradient and single layer centrifugation through silica colloids. Reprod Domest Anim 44: 512-517. http://dx.doi.org/10.1111/j.1439-0531.2008.01265.x
Morrell JM, Johannisson A, Dalin AM, Rodríguez-Martínez H, 2009b. Single-layer centrifugation with Androcoll-E can be scaled up to allow large volumes of stallion ejaculate to be processed easily. Theriogenology 72: 879-884. http://dx.doi.org/10.1016/j.theriogenology.2009.05.015
Morrell JM, Johannisson H, Strutz H, Dalin AM, Rodríguez-Martínez H, 2009c. Colloidal centrifugation of stallion semen: changes in sperm motility, velocity, and chromatin integrity during storage. J Equine Vet Sci 29: 24-32. http://dx.doi.org/10.1016/j.jevs.2008.11.006
Morrell JM, Mari G, Kútvölgyi G, Meurling S, Mislei B, Iacono E, Rodríguez-Martínez H, 2010a. Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single layer centrifugation with Androcoll-E. Reprod Dom Anim 46: 642-645. http://dx.doi.org/10.1111/j.1439-0531.2010.01721.x
Morrell JM, Rodríguez-Martínez H, Johannisson A, 2010b. Single layer centrifugation of stallion spermatozoa improves sperm quality compared with sperm washing. Reprod Biomed 21: 429-436. http://dx.doi.org/10.1016/j.rbmo.2010.03.027
Morrell JM, Klein C, Lundeheim N, Erol E, Troedsson MHT, 2014a. Removal of bacterial from stallion semen by colloid centrifugation. Anim Reprod Sci 145: 47-53. http://dx.doi.org/10.1016/j.anireprosci.2014.01.005
Morrell JM, Stuhtmann G, Meurling S, Lundgren A, Winblad C, Macías-García B, Johannisson A, 2014b. Sperm yield after single layer centrifugation with Androcoll-E is related to the potential fertility of the original ejaculate. Theriogenology 81: 1005-1011. http://dx.doi.org/10.1016/j.theriogenology.2014.01.013
Ortega-Ferrusola C, González-Fernández L, Muriel A, Macías-García B, Rodríguez-Martínez H, Tapia JA, Peña FJ, 2009. Does the microbial flora in the ejaculate affect the freezeability of stallion sperm? Reprod Domest Anim 44: 518-522. http://dx.doi.org/10.1111/j.1439-0531.2008.01267.x
Sieme H, Katila T, Klug E, 2004. Effect of semen collection practices on sperm characteristics before and after storage and on fertility of stallions. Theriogenology 61: 769-784. http://dx.doi.org/10.1016/S0093-691X(03)00251-6
Tischner M, Kosiniak K, Bielanski W, 1974. Analysis of pattern of ejaculation in stallions. J Reprod Fertil 41: 329-335. http://dx.doi.org/10.1530/jrf.0.0410329
Valsa J, Skandhan KP, Khan PS, Sumangala B, Gondalia M, 2012. Split ejaculation study: semen parameters and calcium and magnesium in seminal plasma. Cent European J Urol 65: 216-218. http://dx.doi.org/10.5173/ceju.2012.04.art7
Varner DD, 2008. Developments in stallion semen evaluation. Theriogenology 70: 448-462. http://dx.doi.org/10.1016/j.theriogenology.2008.04.023
© CSIC. Manuscripts published in both the print and online versions of this journal are the property of the 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) licence. You may read the basic information and the legal text of the licence. The indication of the CC BY 4.0 licence must be expressly stated in this way when necessary.
Self-archiving in repositories, personal webpages or similar, of any version other than the final version of the work produced by the publisher, is not allowed.