Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2022-0056
Animal Reproduction (AR)
ORIGINAL ARTICLE

Action of swim-up and caffeine on equine frozen sperm

Natália de Castro Alves; Soraia de Araújo Diniz; Rodrigo Novaes Viegas; Ana Luiza Arigoni; Marina Morra Freitas; Ângela Quintão Lana; Monique de Albuquerque Lagares

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Abstract

Cryopreservation of equine semen is crucial to semen commercialization. However, it reduces sperm motility and longevity. Thus, sperm selection methods and addition of motility-activating substances to sperm, such as caffeine, may improve sperm quality of equine frozen semen. The objective of the current work was to evaluate the effects of caffeine on recovery and quality parameters of frozen-thawed sperm subjected to swim-up selection to be used in intracytoplasmic sperm injection (ICSI) in assisted reproductive techniques. Stallion semen were frozen and after thawing different caffeine concentrations were added to the samples performing four treatments control (no caffeine), 3, 5, and 7.5 mM caffeine. Sperm kinematic and motility were assessed by computer-assisted sperm analysis (CASA). Then, the four treated samples were submitted to the swim-up sperm selection, and the number of recovered sperm and morphology were evaluated at four times 20, 40, 60, and 80 min. The swim-up increased the recovery proportion of normal morphology sperm without (80.1±1%) or with caffeine addition (3mM: 81.2±1%, 5mM: 79.9±1% and 7.5 mM 78.9±1%) compared to the thawed semen (70±2%). However, the addition of 5 mM caffeine induced an increase in sperm motility (38.9±2.8 vs. 32.6±3.4%, P<0.05), and sperm recovery after swim-up (7.9x106 vs. 3.4x106 sperm/ml, P<0.05) compared to the control. The addition of 5 mM caffeine to frozen-thawed equine semen before swim-up selection improved sperm motility and increased the sperm recovery rate while not decreasing the percentage of morphologically normal sperm. Thus, caffeine addition to frozen-thawed equine semen before swim-up selection has potential clinical application in improving sperm quality for use in ICSI.

Keywords

spermatozoa, cryopreservation, freezing, sperm selection, stallion

References

Alves NC, Diniz SA, Viegas RN, Cortes SF, Costa ED, Freitas MM, Martins-Filho OA, Araújo MSS, Lana AMQ, Wenceslau RR, Lagares MA. Addition of caffeine to equine thawed sperm increases motility and decreases nitrite concentration. Andrologia. 2021;53(2):e13918. http://dx.doi.org/10.1111/and.13918. PMid:33368488.

Brandão AC, Arruda RP, Madureira EH, Martins JFP, Assumpção MEOA, Visintin JA. Influência do glicerol e etilenoglicol e da criopreservação sobre o complexo DNA-proteína de espermatozoides em garanhões. Braz J Vet Res Anim Sci. 2006;43(Suppl):68-73. http://dx.doi.org/10.11606/issn.1678-4456.bjvras.2006.26537.

Carrington JL, Carrington AC, Splan RK, Porr CA, Brooks RM. Effect of caffeine, pentoxifylline, and taurine on post-thaw parameters of equine frozen semen. J Equine Vet Sci. 2011;31(5-6):325-26. http://dx.doi.org/10.1016/j.jevs.2011.03.162.

Choi Y-H, Velez IC, Macías-García B, Riera FL, Ballard CS, Hinrichs K. Effect of clinically-related factors on in vitro blastocyst development after equine ICSI. Theriogenology. 2016;85(7):1289-96. http://dx.doi.org/10.1016/j.theriogenology.2015.12.015. PMid:26777560.

Colégio Brasileiro de Reprodução Animal – CBRA. Manual para exame andrológico e avaliação de sêmen animal. 3rd ed. Belo Horizonte: CBRA; 2013.

Colleoni S, Duchi R, Barbacini S, Necchi D, Mari G, Spinaci M, Lazzari G, Galli C. Practical applications of OPU, ICSI and IVC in equine reproduction. Reprod Domest Anim. 2009;44:62.

Colleoni S, Lagutina I, Lazzari G, Rodriguez-Martinez H, Galli C, Morrell JM. New methods for selecting stallion spermatozoa for assisted reproduction. J Equine Vet Sci. 2011;31(9):536-41. http://dx.doi.org/10.1016/j.jevs.2011.03.009.

Correa JR, Zavos PM. Preparation and recovery of frozen-thawed bovine spermatozoa via various sperm selection techniques employed in assisted reproductive technologies. Theriogenology. 1996;46(7):1225-32. http://dx.doi.org/10.1016/S0093-691X(96)00293-2. PMid:16727985.

Galli G, Duchi R, Crotti G, Turini P, Ponderato N, Colleoni S, Lagutina I, Lazzari G. Bovine embryo technologies. Theriogenology. 2003;59(2):599-616. http://dx.doi.org/10.1016/S0093-691X(02)01243-8. PMid:12499007.

García BM, Fernández LG, Morrell JM, Ferrusola CO, Tapia JA, Martínez HR, Peña FJ. Single-layer centrifugation through colloid positively modifies the sperm subpopulation structure of frozen-thawed stallion spermatozoa. Reprod Domest Anim. 2009;44(3):523-6. http://dx.doi.org/10.1111/j.1439-0531.2008.01276.x. PMid:18992085.

Henkel R. Sperm preparation: state-of-the-art--physiological aspects and application of advanced sperm preparation methods. Asian J Androl. 2012;14(2):260-9. http://dx.doi.org/10.1038/aja.2011.133. PMid:22138904.

Hoogewijs M, Piepers S, Govaere J, Schauwer C, Kruif A, Morrell JM. Sperm longevity following pre-freeze sperm selection. J Equine Vet Sci. 2012;32(8):489. http://dx.doi.org/10.1016/j.jevs.2012.06.048.

Huo L-J, Ma X-H, Yang Z-M. Assessment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage. Theriogenology. 2002;58(7):1349-60. http://dx.doi.org/10.1016/S0093-691X(02)00953-6. PMid:12387348.

Kenney RM, Bergman RV, Cooper WL, Morse FW. Minimal contamination techniques for breeding mares: techniques and preliminary findings. Proceedings of American Association of Equine Practitioners. 1975;21:327-36.

Kim EK, Kim EH, Kim EA, Lee KA, Shin JE, Kwon H. Comparison of the effect of different media on the clinical outcomes of the density-gradient centrifugation/swim-up and swim-up methods. Clin Exp Reprod Med. 2015;42(1):22-9. http://dx.doi.org/10.5653/cerm.2015.42.1.22. PMid:25874170.

Landim-Alvarenga F, Fernandes C, Devito L, Blanco ID, Alvarenga M. New assisted reproductive technologies applied to the horse industry: successes and limitations. Anim Reprod. 2008;5:67-82.

Larentis GR, Bastos HBA. Equine sperm selection. Clin Res Anim Sci. 2019;1(1):CRAS.000501.

Lazzari G, Crotti G, Turini P, Duchi R, Mari G, Zavaglia G, Barbacini S, Galli C. Equine embryos at the compacted morula and blastocyst stage can be obtained by intracytoplasmic sperm injection (ICSI) of in vitro matures oocytes with frozen-thawed spermatozoa from semen of different fertilities. Theriogenology. 2002;58(2-4):709-12. http://dx.doi.org/10.1016/S0093-691X(02)00777-X.

Maia MS, Bicudo SD. Radicais livres, antioxidantes e função espermática em mamíferos: uma revisão. Rev Bras Reprod Anim. 2009;33:183-93.

Mbizvo MT, Johnston RC, Baker GH. The effect of the motility stimulants, caffeine, pentoxifylline, and 2-deoxyadenosine on hyperactivation of cryopreserved human sperm. Fertil Steril. 1993;59(5):1112-7. http://dx.doi.org/10.1016/S0015-0282(16)55937-8. PMid:8486183.

Mies A Fo. Reprodução dos animais e inseminação artificial. 3rd ed. Porto Alegre: Sulina; 1975. Tecnologia do sêmen I – exame e classificação; p. 423-58.

Morrell JM, Dalin AM, Rodriguez-Martinez H. Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival. Equine Vet J. 2009;41(1):53-8. http://dx.doi.org/10.2746/042516408X322139. PMid:19301582.

Morrell JM. Stallion sperm selection: past, present and future trends. J Equine Vet Sci. 2012;32(8):436-40. http://dx.doi.org/10.1016/j.jevs.2012.05.069.

Muratori M, Tarozzi N, Carpentiero F, Danti S, Perrone FM, Cambi M, Casini A, Azzari C, Boni L, Maggi M, Borini A, Baldi E. Sperm selection with density gradient centrifugation and swim up: effect on DNA fragmentation in viable spermatozoa. Sci Rep. 2019;9(1):7492. http://dx.doi.org/10.1038/s41598-019-43981-2. PMid:31097741.

Navarro-Serna S, París-Oller E, Simonik O, Romar R, Gadea J. Replacement of albumin by preovulatory oviductal fluid in swim-up sperm preparation method modifies boar sperm parameters and improves in vitro penetration of oocytes. Animals. 2021;11(5):1202. http://dx.doi.org/10.3390/ani11051202. PMid:33922134.

Pariz JR, Hallak J. Effects of caffeine supplementation in post‐thaw human semen over different incubation periods. Andrologia. 2016;48(9):961-6. http://dx.doi.org/10.1111/and.12538. PMid:26781217.

Pérez-Cerezales S, Laguna-Barraza R, Castro AC, Sánchez-Calabuig MJ, Cano-Oliva E, Castro-Pita FJ, Montoro-Buils L, Pericuesta E, Fernández-González R, Gutiérrez-Adán A. Sperm selection by thermotaxis improves ICSI outcome in mice. Sci Rep. 2018;8(1):2902. http://dx.doi.org/10.1038/s41598-018-21335-8. PMid:29440764.

Pillet E, Batellier F, Duchamp G, Furstoss V, Le Vern Y, Kerboeuf D, Vidament M, Magistrini M. Freezing stallion semen in INRA96-based extender improves fertility rates in comparison with INRA82. Dairy Sci Technol. 2008;88(2):257-65. http://dx.doi.org/10.1051/dst:2008002.

Podico G, Ellerbrock RE, Curcio BR, Cheong SH, Lima FS, Canisso IF. Single-layer colloid centrifugation as a method to process urine-contaminated stallion semen after freezing-thawing. J Equine Vet Sci. 2020;87:102910. http://dx.doi.org/10.1016/j.jevs.2020.102910. PMid:32172909.

Rappa KL, Rodriguez HF, Hakkarainen GC, Anchan RM, Mutter GL, Asghar W. Sperm processing for advanced reproductive technologies: where are we today? Biotechnol Adv. 2016;34(5):578-87. http://dx.doi.org/10.1016/j.biotechadv.2016.01.007. PMid:26845061.

Roca J, Martinez-Alborcia MJ, Gil MA, Parrilla I, Martinez EA. Dead spermatozoa in raw semen samples impair in vitro fertilization outcomes of frozen-thawed spermatozoa. Fertil Steril. 2013;100(3):875-81. http://dx.doi.org/10.1016/j.fertnstert.2013.05.020. PMid:23768987.

Rodríguez-Martinez H, Larsson B, Pertoft H. Evaluation of sperm damage and techniques for sperm clean up. Reprod Fertil Dev. 1997;9(3):297-308. http://dx.doi.org/10.1071/R96081. PMid:9261878.

Squires EL, Pickett BW, Graham JK, Vanderwall DK, McCue PM, Bruemmer JE. Cooled and frozen stallion semen. Fort Collins: Colorado State University; 1999. Animal Reproduction and Biotechnology Laboratory Bull. No. 09.

Stephens TD, Brooks RM, Carrington JL, Cheng L, Carrington AC, Porr CA, Splan RK. Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. J Equine Vet Sci. 2013;33(8):615-21. http://dx.doi.org/10.1016/j.jevs.2012.10.004.

Tash JS, Mann FRS. Adenosine 3:5´-cyclic monophosphate in relation to motility and senescence of spermatozoa. Proc R Soc Lond B Biol Sci. 1973;184(1074):109-14. http://dx.doi.org/10.1098/rspb.1973.0036. PMid:4147947.

Volpes A, Sammartano F, Rizzari S, Gullo S, Marino A, Allegra A. The pellet swim-up is the best technique for sperm preparation during in vitro fertilization procedures. J Assist Reprod Genet. 2016;33(6):765-70. http://dx.doi.org/10.1007/s10815-016-0696-2. PMid:26984108.

Voss JL. Breeding efficiency. In: McKinnon AO, Voss JL, editors. Equine reproduction. Philadelphia: Lea & Febiger; 1993. p. 705-15.

Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci. 2000;60-61:481-92. http://dx.doi.org/10.1016/S0378-4320(00)00099-3. PMid:10844218.
 


Submitted date:
05/31/2022

Accepted date:
10/27/2022

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