Camilo Hernández-Avilés, Jorge Zambrano-Varón, Claudia Jiménez-Escobar



Semen evaluation is an important component for the assessment of the stallion breeding potential, as well as for the diagnosis of subfertility/infertility cases. Furthermore, accurate estimation of the damage suffered by the sperm cell after cooling or freezing procedures is necessary for the development of newer procedures to maintain sperm integrity and function. Nevertheless, commonly used methods for sperm quality evaluation (sperm motility or sperm morphology) are not completely associated with the fertilizing potential of the spermatozoa under in vitro conditions, and in the best-case scenario are poor- to- moderately associated with in vivo fertility. In recent years, the introduction of advanced methods based on the use of fluorochromes for sperm evaluation has improved the clinician’s and researcher’s capacity to account for the differences on the fertility potential between sires, as well as critically evaluate the effect of several methods to preserve stallion sperm. The aim of this paper was to review some of the current fluorescence-based methods used for the evaluation of equine semen as an alternative for the selection of breeding stallions, the diagnosis of subfertility/infertility, and the estimation of optimal protocols for sperm preservation under laboratory conditions.



La evaluación del semen es un componente importante en  la evaluación del potencial reproductivo de sementales, así como para el diagnóstico de casos de subfertilidad / infertilidad. Además, es necesario realizar una estimación precisa del daño sufrido por la célula espermática después de los procedimientos de refrigeración o criopreservación para el desarrollo de nuevos procedimientos a fin de mantener la integridad y la función del espermatozoide. Sin embargo, los métodos comúnmente utilizados para la evaluación de la calidad espermática (motilidad o morfología espermática) no están completamente asociados con el potencial de fertilización del espermatozoide en condiciones in vitro, y en el mejor de los casos están poco a moderadamente asociados con la fertilidad in vivo . En los últimos años, la introducción de métodos avanzados basados en el uso de fluorocromos para la evaluación espermática ha mejorado la capacidad del clínico y del investigador para explicar las diferencias en el potencial de fertilidad entre los sementales, así como evaluar críticamente el efecto de varios métodos para preservar el espermatozoide equino. El objetivo de este trabajo fue revisar algunos de los métodos actuales basados en fluorescencia que están siendo  utilizados para la evaluación del semen equino como una alternativa para la selección de sementales reproductores, el diagnóstico de subfertilidad / infertilidad y la estimación de protocolos óptimos para la preservación del espermatozoide bajo condiciones de laboratorio.

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Aitken RJ, Gibb Z, Baker MA, Drevet J, Gharagozloo P. 2016. Causes and consequences of oxidative stress in spermatozoa. Reproduction Fertility Development 28(1-2):1-10.

Aitken RJ, Whiting S, De luliis GN, McClymont S, Mitchell LA, Baker MA. 2012. Electrophilic aldehydes generated by sperm metabolism activate mitochondrial reactive oxygen species and apoptosis by targeting succinate dehydrogenase. J Biol Chem 287:33048-33060.

Aitken RJ. 2017. Reactive oxygen species as mediators of sperm capacitation and pathological damage. Molecular Reproduction and Development 84(10): 1039-1052.

Alvarez JG, Storey BJ. 1984. Assessment of cell damage caused by spontaneous lipid peroxidation in rabbit spermatozoa. Biology of Reproduction 30 (2):833-841.

Amann RP, Hammerstedt RH. 1993. In vitro evaluation of sperm quality: An opinion. Journal of Andrology 14:397-406.

Amann RP, Waberski D. 2014. Computer-assisted sperm analysis (CASA): Capabilities and potential development. Theriogenology 81(1):5-17.

Balao da Silva C, Ortega-Ferrusola C, Morrell JM, Rodríguez-Martínez H, Peña FJ. 2016. Flow cytometric chromosomal sex sorting of stallion spermatozoa induces oxidative stress on mitochondria and genomic DNA. Reproduction in Domestic Animals 51(1):18-25.

Ball BA, Vo A. 2002. Detection of lipid peroxidation in equine spermatozoa based upon the lipophilic fluorescent dye C11-BODIPI 581/591. Journal of Andrology 23(2):259-269.

Ball L, Ott RS, Mortimer RG, Simons JC. 1983. Manual for breeding soundness examination of bulls. Journal of the Society for Theriogenology 12:1-65.

Barrier-Battut I, Kempfer A, Becker J, Lebailly L, Camugli S, Chevrier L. 2016. Development of a new fertility prediction model for stallion semen, including flow cytometry. Theriogenology 86:111-1131.

Barrier-Battut I, Kempfer A, Lemasson N, Chevrier L, Camugli S. 2017. Prediction of the fertility of stallion frozen-thawed semen using a combination of computer-assisted motility analysis, microscopical observation and flow cytometry. Theriogenology 97:186-200.

Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC. 2000. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. Journal of Andrology 21(6):895-902.

Baumber J, Ball BA, Linfor JJ, Meyers SA. 2003a. Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa. Journal of Andrology 24(4):621-628.

Baumber J, Sabeur K, Vo A, Ball BA. 2003b. Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. Theriogenology 60(7):1239-1247.

Bedford SJ, Varner DD, Meyers SA. 2000. Effects of cryopreservation on the acrosomal status of stallion spermatozoa. Journal of Reproduction and Fertility Suppl 56:133-140.

Bedford-Guaus SJ, McPartlin LA, Varner DD. 2012. Characterization of equine Phospholipase C Zeta: A review and preliminary results on expression defects in subfertile stallions. Journal of Equine Veterinary Science 32:445-450.

Boerke A, Tsai PS, Garcia-Gil N, Brewis IA, Gadella BM. 2008. Capacitation-dependent reorganization of microdomains in the apical sperm head plasma membrane: functional relationship with zona binding and the zona-induced acrosome reaction. Theriogenology 70:1188-1196.

Bosard T, Love CC, Brinsko SP, Blanchard TL, Thompson J, Varner DD. 2005. Evaluation and diagnosis of acrosome function/dysfunction in the stallion. Animal Reproduction Science 89(1-4):215-217.

Brinkerhoff JM, Love CC, Thompson JA, Blodgett G, Teague SR, Varner DD. 2010. Influence of mare age, pre-breeding mare status, breeding methods, and stallion on first cycle pregnancy rates on a large commercial breeding farm. Animal Reproduction Science 121:159.

Brinsko SP, Love CC, Bauer JE, Macpherson ML, Varner DD. 2007. Cholesterol-to-phospholipid ratio in whole sperm and seminal plasma from fertile stallions and stallions with unexplained subfertility. Animal Reproduction Science 99:65-71.

Brito LF, Greene LM, Kelleman A, Knobbe M, Turner R. 2011. Effect of method and clinician on stallion sperm morphology evaluation. Theriogenology 76: 745-750.

Bueno VLC, Bastos HBA, Centeno LAM, Kretzmann NA, Bertolini M, Mattos RC, Rechsteiner FS. 2018. The role of PLCζ and WPB2NL gene expression in semen quality and fertility of stallions. Journal of Equine Veterinary Science 66:33-34.

Burnaugh L, Ball BA, Sabeur K, Thomas AD, Meyers SA. 2010. Osmotic stress stimulates generation of superoxide anion by spermatozoa in horses. Animal Reproduction Science 117(3-4):249-260.

Burnaugh L, Sabeur K, Ball BA. 2007. Detection of superoxide anion by equine spermatozoa as detected by dihydroethidium. Theriogenology 67: 580-589.

Burruel V, Klooster KL, Chitwood J, Ross PJ, Meyers SA. 2013. Oxidative damage to rhesus macaque spermatozoa results in mitotic arrest and transcript abundance changes in early embryos. Biology of Reproduction 89(3):72.

Casey PJ, Hillman RB, Robertson KR, Yudin AI, Liu IK, Drobnis EZ. 1993. Validation of an acrosomal stain for equine sperm that differentiates between living and dead sperm. Journal of Andrology 14(4):289-297.

Cheng FP, Fazeli A, Voorhout WF, Marks A, Bevers MM, Colenbrander B. 1996. Use of peanut agglutinin to assess the acrosomal status and the zona pellucida-induced acrosome reaction in stallion spermatozoa. Journal of Andrology 17:674-682.

Choi YH, Varner DD, Love CC, Hartman DL, Hinrichs K. 2011. Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse. Reproduction 142(4):529-538.

Colenbrander B, Gadella BM, Stout TA. 2003. The predictive value of semen analysis in the evaluation of stallion fertility. Reproduction in Domestic Animals 38(4):305-311.

Darr CR, Cortopassi GA, Datta S, Varner DD, Meyers SA. 2016a. Mitochondrial oxygen consumption is a unique indicator of stallion spermatozoal health and varies with cryopreservation media. Theriogenology 86(5):1382-1392.

Darr CR, Varner DD, Teague S, Cortopassi GA, Datta S, Meyers SA. 2016b. Lactate and pyruvate are major sources of energy for stallion sperm with dose effects on mitochondrial function, motility and ROS production. Biology of Reproduction 95(2):34.

Drobnis EZ, Crowe LM, Berger T, Anchordoguy TJ, Overstreet JW, Crowe JH. 1993. Cold shock damage is due to lipid phase transitions in cell membranes: a demonstration using sperm as a model. Journal of Experimental Zoology 265(4):432-437.

Eddy EM, O´Brien DA. 1994. The Spermatozoon. In: Knobil E, Neill JD (Eds.), The Physiology of Reproduction. 2nd edition. Raven Press, NY. pp: 29-77.

Ertmer F, Oldenhof H, Schütze S, Rohn K, Wolkers WF, Sieme H. 2017. Induced sub-lethal oxidative damage affects osmotic tolerance and cryosurvival of spermatozoa. Reproduction Fertility Development 29(9):1739-1750.

Evenson DP, Darzynkiewicz Z, Melamed MR. 1980. Relation of mammalian sperm chromatin heterogeneity to fertility. Science 210:1131.

Evenson DP, Jost LK, Varner DD. 2000. Relationship between sperm nuclear protamine free-SH status and susceptibility to DNA denaturation. Journal of Reproduction and Fertility Suppl 56:401.

Evenson DP. 2016. The Sperm Chromatin Structure Assay (SCSA®) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility. Animal Reproduction Science 169:56-75.

Farlin ME, Jasko DJ, Graham JK, Squires EL. 1992. Assessment of Pisum sativum agglutinin in identifying acrosomal damage in stallion spermatozoa. Molecular Reproduction and Development 32:23-27.

Fatehi AN, Bevers MM, Schoevers E, Roelen BA, Colenbrander B, Gadella BM. 2006. DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after the first cleavages. Journal of Andrology 27(2):176-188.

Foster ML, Love CC, Varner DD, Brinsko SP, Hinrichs K, Teague S, Lacaze K, Blanchard TL. 2011. Comparison of methods for assessing integrity of equine sperm membranes. Theriogenology 76:334-341.

Gadella BM. 2008. Sperm membrane physiology and relevance for fertilization. Animal Reproduction Science 107:229-236.

Galantino-Homer HL, Visconti PE, Kopf GS. 1997. Regulation of protein tyrosine phosphorylation during bovine sperm capacitation by a cyclic adenosine 3’5’-monophosphate-dependent pathway. Biology of Reproduction 58(3):707-719.

Gallardo-Bolaños JM, Miró-Morán A, Balao da Silva CM, Morillo-Rodríguez A, Plaza-Dávila M, Aparicio IM, Tapia JA, Ortega-Ferrusola C, Peña FJ. 2014. Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration. PLoS One 7: e30688.

Garner DL, Johnson LA. 1995. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biology of Reproduction 53:276-284.

Gibb Z, Lambourne SR, Aitken RJ. 2014. The paradoxical relationship between stallion fertility and oxidative stress. Biology of Reproduction 91(3):77.

Gibb Z, Lambourne SR, Quadrelli J, Smith ND, Aitken RJ. 2015. L-carnitine and pyruvate are prosurvival factors during the storage of stallion spermatozoa at room temperature. Biology of Reproduction 93(4):104.

Gibb Z, Lambourne SR, Curry BJ, Hall SE, Aitken RJ. 2016. Aldehyde dehydrogenase plays a pivotal role in the maintenance of stallion sperm motility. Biology of Reproduction 94(6):133.

Gonzalez-Castro RA, Graham JK, Carnevale EM. 2018. Localization and quantitative expression of Phospholipase C Zeta in equine sperm using commercial antibodies. Reproduction Fertility Development 30(1):228.

González-Fernández L, Macías-García B, Loux SC, Varner DD, Hinrichs K. 2013. Focal adhesion kinases and calcium/calmodulin dependent protein kinases regulate protein tyrosine phosphorylation in stallion sperm. Biology of Reproduction 88(6):138.

Gravance CG, Garner DL, Baumber J, Ball BA. 2000. Assessment of equine sperm mitochondrial function using JC-1. Theriogenology 53(9):1691-1703.

Gravance CG, Liu IK, Davis RO, Hughes JP, Casey PJ. 1996. Quantification of normal head morphometry of stallion spermatozoa. Journal of Reproduction and Fertility 108(1):41-46.

Hernández-Avilés C, Gómez-Romero M, Buitrago-Horta R, Lozano-Márquez H, Jiménez-Escobar C, Zambrano-Varón J. 2018a. Evaluation of post-thaw sperm function and integrity parameters under different freezing regimens in Colombian Paso Fino stallions. Journal of Equine Veterinary Science 67:7-14.

Hernández-Avilés C, Love CC, Serafini R, Ghosh S, Teague SR, La Caze KA, Varner DD. 2018b. The effects of extender glucose concentration and storage temperature on stallion semen quality following cooled storage. Journal of Equine Veterinary Science 66:46.

Heytens E, Parrington J, Coward K, Young C, Lambrecht S, Yoon SY, Fissore RA, Hamer R, Deane CM, Ruas M, Grasa P, Soleimani R, Cuvelier CA, Gerris J, Dhont M, Deforce D, Leybaert L, De Sutter P. 2009. Reduced amounts and abnormal forms of phospholipase C zeta (PLCζ) in spermatozoa from infertile men. Human Reproduction 24:2417-2428.

Hinrichs K. 2018. Assisted reproductive techniques in mares. Reproduction in Domestic Animals 53(Suppl. 2):4-13.

Hoogewijs MK, de Vlieguer SP, Govaere JL, de Schauwer C, de Kruif A, van Soom A. 2012. Influence of counting chamber type on CASA outcomes of equine semen analysis. Equine Veterinary Journal 44(5):542-549.

Hopkins FM, Spitzer JC. 1997. The new society for theriogenology breeding soundness evaluation system. Veterinary Clinics North America: Food Animal Practice 13:283-293.

Hurtgen JP. 1992. Evaluation of the stallion for Breeding soundness. Veterinary Clinics North America: Equine Practice 8(1):149-165.

Janson-Whitesell M, McDonnell SM, Turner RM. 2014. Modernizing the stallion breeding soundness evaluation. Journal of Equine Veterinary Science 34:50-51.

Jasko DJ, Lein DH, Foote RH. 1990. Determination of the relationship between sperm morphologic classifications and fertility in stallions: 66 cases (1987-1998). Journal of the American Veterinary Medical Association 197(3):389-394.

Jasko DJ, Little TV, Lein DH, Foote RH. 1992. Comparison of spermatozoa movement and semen characteristics with fertility in stallions: 64 cases (1987-1988). Journal of the American Veterinary Medical Association 200:979-985.

Kenney RM, Evenson DP, Garcia MC, Love CC. 1995. Relationships between sperm chromatin structure, motility, and morphology of ejaculated sperm, and seasonal pregnancy rate. Biology of Reproduction 52 (Issue Monograph Series 1): 647-653.

Kenney RM, Hurtgen J, Pierson R, Witherspoon D, Simons J. 1983. Theriogenology and the equine, Part II: The stallion. Society for Theriogenology Manual for Clinical Fertility Evaluation of the Stallion. Hastings, NE.

Kiser AM, Brinsko SP, Love CC, Varner DD, Sudderth K, Blanchard TL. 2014. Relationship of sperm quality to fertility after 4 days of cooled storage of equine semen. Journal of Equine Veterinary Science 34: 602-605.

Kuisma P, Andersson M, Koskinen E, Katila T. 2006. Fertility of frozen-thawed stallion semen cannot be predicted by the currently used laboratory methods. Acta Veterinaria Scandinavica 17(48):14.

Leemans B, Gadella BM, Sostaric E, Nelis H, Stout TA, Hoogewijs M, Van Soom A. 2014. Oviduct binding and elevated environmental pH induce protein tyrosine phosphorylation in stallion spermatozoa. Biology of Reproduction 91(1):13.

Linfor JJ, Meyers SA. 2002. Detection of DNA damage in response to cooling injury in equine spermatozoa using single-cell gel electrophoresis. Journal of Andrology 23 (1):107-113.

Loomis PR, Graham JK. 2008. Commercial semen freezing: Individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Animal Reproduction Science 105(1-2):119-128.

Love CC, Noble JK, Standridge SA, Bearden CT, Blanchard TL, Varner DD, Cavinder CA. 2015. The relationship between sperm quality in cool-shipped semen and embryo recovery rate in horses. Theriogenology 84:1587-1593.

Love CC, Thompson JA, Brinsko SP, Rigby SL, Blanchard TL, Lowry VK, Varner DD. 2003. Relationship between stallion sperm motility and viability as detected by two fluorescence staining techniques using flow cytometry. Theriogenology 60(6):1127-1138.

Love CC, Thompson JA, Lowry VK, Varner DD. 2002. Effect of storage time and temperature on stallion sperm DNA and fertility. Theriogenology 57(3):1135-1142.

Love CC, Varner DD, Thompson JA. 2000. Intra- and inter-stallion variation in sperm morphology and their relationship with fertility. Journal of Reproduction and Fertility Suppl 56: 93-100.

Love CC. 2005. The sperm chromatin structure assay: a review of clinical applications. Animal Reproduction Science 89(1-4):39-45.

Love CC. 2011. Relationship between sperm motility, morphology and the fertility of stallions. Theriogenology 76(3):547-557.

Love CC. 2012. Measurement of concentration and viability in stallion sperm. Journal of Equine Veterinary Science 32(8):464-466.

Love CC. 2018. Sperm quality assays: How good are they? The horse perspective. Animal Reproduction Science 194:63-70.

Love CC. Brinsko SP, Rigby SL, Thompson JA, Blanchard TL, Varner DD. 2005. Relationship of seminal plasma level and extender type to sperm motility and DNA integrity. Theriogenology 63:1584-1591.

Macías-García B, González-Fernández L, Loux SC, Rocha AM, Guimaraes T, Peña FJ, Varner DD, Hinrichs K. 2015. Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm. Reproduction 149: 87-99.

Macías-García B, Miró-Morán A, González-Fernández L, Ortega-Ferrusola C, Morillo-Rodríguez A, Bolaños JM, da Silva CM, Martínez HR, Tapia JA, Peña FJ. 2012. The mitochondria of stallion spermatozoa are more sensitive than the plasmalemma to osmotic-induced stress: role of c-Jun N-terminal kinase (JNK) pathway. Journal of Andrology 33 (1):105-113.

Martínez-Pastor F, Mata-Campuzano M, Álvarez-Rodríguez M, Álvarez M, Anel L, de Paz P. 2010. Probes and techniques for sperm evaluation by flow cytometry. Reproduction in Domestic Animals 45(Suppl. 2):67-78.

Martín-Muñoz P, Anel-López L, Ortiz-Rodríguez JM, Álvarez M, de Paz P, Balcao da Silva C, Rodríguez-Martinez R, Gil MC, Anel L, Peña FJ, Ortega-Ferrusola C. 2018. Redox cycling induces spermptosis and necrosis in stallion spermatozoa while the hydroxyl radical (OH●) only induces spermptosis. Reproduction in Domestic Animals 53(1):54-67.

Martín-Muñoz P, Ortega-Ferrusola C, Vizuete G, Plaza-Dávila M, Rodríguez-Martínez, Peña FJ. 2015. Depletion of intracellular thiols and increased production of 4-hydroxynonenal that occur during cryopreservation of stallion spermatozoa lead to caspase activation, loss of motility and cell death. Biology of Reproduction 93:1-11.

McPartlin LA, Littell J, Mark E, Nelson JL, Travis AJ, Bedford-Guaus SJ. 2008. A defined medium supports changes consistent with capacitation in stallion sperm, as evidenced by increases in protein tyrosine phosphorylation and high rates of acrosomal exocytosis. Theriogenology 69(5):639-650.

Merkies K, Chenier T, Plante C, Buhr MM. 2000. Assessment of stallion spermatozoa viability by flow cytometry and light microscope analysis. Theriogenology 54:1215-1224.

Meyers S, Liu IKM, Overstreet JW, Drobnis EZ. 1996. Sperm-zona pellucida binding and zona-induced acrosome reaction in the horse: comparisons between fertile and subfertile males. Theriogenology 46:1277-1288.

Meyers SA, Overstreet JW, Liu IK, Drobnis EZ. 1995. Capacitation in-vitro of stallion spermatozoa: comparison of progesterone-induced acrosome reactions in fertile and subfertile males. Journal of Andrology 16:47-54.

Miró-Arias M, Gallardo-Bolaños JM, Morillo-Rodríguez A, Balao da Silva C, Aparicio IM, Tapia JA, Valecillo A, Vega-Pla JL, Peña FJ, Ortega-Ferrusola C. 2013. Sperm susceptibility to Poxidative stress in the Retuertas endangered horse. Journal of Equine Veterinary Science 33:962-968.

Murcia-Robayo RY, Jouanisson E, Beauchamp G, Diaw M. 2018. Effects of staining method and clinician experience on the evaluation of stallion sperm morphology. Animal Reproduction Science 188:165-169.

Neild DM, Brouwers JF, Colenbrander B, Agüero A, Gadella BM. 2005. Lipid peroxide formation in relation to membrane stability of fresh and frozen thawed stallion spermatozoa. Molecular Reproduction and Development 72(2):230-238.

Nomikos M, Elgmati K, Theodoridou M, Calver BL, Cumbes B, Nounesis G, Swann K, Lai FA. 2011. Male infertility-linked point mutation disrupts the Ca2+ oscillation-inducing and PIP2 hydrolysis activity of sperm PLC zeta. Biochemical Journal 434: 211-217.

Ortega-Ferrusola C, González-Fernández L, Morrell JM, Salazar-Sandoval C, Macías-García B, Rodríguez-Martinez H, Tapia JA, Peña FJ. 2009a. Lipid peroxidation, assessed with BODIPI-C11, increases after cryopreservation of stallion spermatozoa, is stallion-dependent and is related to apoptotic-like changes. Reproduction 138(1):55-63.

Ortega-Ferrusola C, Macías-García B, Gallardo-Bolaños JM, González-Fernández L, Rodríguez-Martínez H, Tapia JA, Peña FJ. 2009b. Apoptotic markers can be used to forecast the freezability of stallion spermatozoa. Animal Reproduction Science 114(4):393-403.

Panarace M, Pellegrini RO, Basualdo MO, Bele M, Ursino DA, Cisterna R, Desimone G, Rodríguez E, Medina MJ. 2014. First field results on the use of stallion sex-sorted semen in a large-scale embryo transfer program. Theriogenology 81(4):520-525.

Parks JE, Lynch DV. 1992. Lipid composition and thermotropic phase behavior of boar, bull, stallion and rooster sperm membranes. Cryobiology 29(2):255-266.

Peña FJ, Ball BA, Squires EL. 2018. A new method for evaluating stallion sperm viability and mitochondrial membrane potential in fixed semen samples. Cytometry Part B: Clinical Cytometry 94(2):302-311.

Peña FJ, Martín-Muñoz P, Ortega-Ferrusola C. 2016. Flow cytometry probes to evaluate stallion spermatozoa. Journal of Equine Veterinary Science 43:S23-S28.

Peña FJ, Plaza Davila M, Ball BA, Squires EL, Martin Muñoz P, Ortega Ferrusola C, Balao da Silva C.2015. The impact of reproductive technologies on stallion mitochondrial function. Reproduction in Domestic Animals 40:529-537.

Petrunkina AM, Waberski D, Günzel-Apel AR, Töpfer-Petersen E. 2007. Determinants of sperm quality and fertility in domestic species. Reproduction 134:3-17.

Plaza-Davila M, Muñoz PM, Bolaños JM, Stout TA, Gadella BM, Tapia JA, da Silva CB, Ortega-Ferrusola C, Peña FJ. 2016. Mitochondrial ATP is required for the maintenance of membrane integrity in stallion spermatozoa, whereas motility requires both glycolysis and oxidative phosphorylation. Reproduction 152 (6): 683–694.

Quintero-Moreno A, Miró J, Teresa-Rigau A, Rodríguez-Gil JE. 2003. Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology 59(9):1973-1990.

Rathi R, Colenbrander B, Bevers MM, Gadella BM. 2001. Evaluation of in vitro capacitation of stallion spermatozoa. Biology of Reproduction 65:462-470.

Rathi R, Colenbrander B, Stout TAE, Gadella BM. 2003. Progesterone induces acrosome reaction in stallion spermatozoa via a protein tyrosine kinase dependent pathway. Molecular Reproduction and Development 64(1):120-128.

Raudsepp T, McCue ME, Das PJ, Dobson L, Vishnoi M, Fritz KL, Schaefer R, Rendahl AK, Derr JN, Love CC, Varner DD, Chowdhary BP. 2012.

Genome-wide association study implicates testis-sperm specific FKBP6 as a susceptibility locus for impaired acrosome reaction in stallions. PLoS Genet 8(12):e1003139.

Sabeur K, Ball BA. 2006. Detection of superoxide anion generation by equine spermatozoa. American Journal of Veterinary Research 67(4):701-706.

Schrimpf R, Dierks C, Martinsson G, Sieme H, Distl O. 2014. Genome-wide association study identifies Phospholipase C Zeta 1 (PLCz1) as a stallion fertility locus in Hanoverian warmblood horses. PLoS One 9(10):e109675.

Serafini R, Varner DD, Bissett W Jr., Blanchard TL, Teague SR, Love CC. 2015. Sperm DNA quality evaluated by comet assay and sperm chromatin structure assay in stallions after unilateral orchiectomy. Theriogenology 84(5):833-837.

Serafini R, Varner DD, Blanchard TL, Teague SR, LaCaze K, Love CC. 2018. Effects of seminal plasma and flash-freezing on DNA structure of stallion epididymal sperm exposed to different potentiators of DNA damage. Theriogenology 117:34-39.

Stump KE, Teague SR, Love CC, Blanchard TL, Brinsko SP, Long CR, Varner DD. 2014. Assessment of stallion sperm membrane integrity using three different flow cytometric methods. Journal of Equine Veterinary Science 34(1):83.

Swann K, Saunders CM, Rogers NT, Lai FA. 2006. PLCζ (zeta): A sperm protein that triggers Ca2+ oscillations and egg activation in mammals. Seminars in Cell and Developmental Biology 17(2):264-73.

Teague SR, Ligon EM, Serafini R, Varner DD, Love CC. 2018. Validation of a fixable stain for assessing the viability of stallion sperm. Journal of Equine Veterinary Science 66:42.

Trentin JM, González-Castro RA, Rubin MIB, Graham JK, Carnevale EM. 2018. Fixable stain for flow cytometric evaluation of stallion sperm viability. Journal of Equine Veterinary Science 66: 70-71.

Uribe P, Villegas JV, Boguen R, Treulen F, Sánchez R, Mallmann P, Isachenko V, Rahimi G, Isachenko E. 2017. Use of the fluorescent dye tetra-methylrhodamine-methyl-ester-perchlorate for mitochondrial membrane potential assessment in human spermatozoa. Andrologia 49:e12753.

Varner DD, Bowen JA, Johnson L. 1993. Effect of heparin on capacitation/acrosome reaction of equine sperm. Archives of Andrology 31:199-207.

Varner DD, Brinsko SP, Blanchard TL, Love CC, Macpherson ML, Heck RS, Johnson L. 2001. Subfertility in stallions associated with spermatozoal acrosome dysfunction. Proceedings of the Annual Convention of the AAEP 47:227-228.

Varner DD, Johnson L. 2007. From a sperm’s eye view – revisiting our perception of this intriguing cell. Proceedings of the Annual Convention of the AAEP 53:104-177.

Varner DD, Thompson JA, Blanchard TL, Heck R, Love CC, Brinsko SP, Johnson L. 2002. Induction of the acrosome reaction in stallion spermatozoa: effects of incubation temperature, incubation time, and ionophore concentration. Theriogenology 58:303-306.

Varner DD. 2016. Approaches to the breeding soundness examination and interpretation or results. Journal of Equine Veterinary Science 43(Suppl.):S37-44.

Veeramachaneni DNR, Moeller CL, Pickett BW, Shiner KA, Sawyer HR. 1993. On processing and evaluation of equine seminal samples for cytopathology and fertility assessment: The utility of electron microscopy. Journal of Equine Veterinary Science 13:207-215.

Visconti PE, Bailey JL, Moore GD, Pan D, Olds-Clarke P, Kopf GS. 1995. Capacitation of mouse spermatozoa. I. Correlation between the capacitation state and protein tyrosine phosphorylation. Development 121(4):1129-1137.

Voss JL, Pickett BW, Squires EL. 1981. Stallion spermatozoal morphology and motility and their relationship with fertility. Journal of the American Veterinary Medical Association 178: 287.

Yesté M, Bonet S, Rodríguez-Gil JE, Rivera del Álamo MM. 2018. Evaluation of sperm motility with CASA-Mot: which factors may influence our measurements? Reproduction Fertility and Development 30(6):789-798.

Yesté M, Estrada E, Rocha LG, Marín H, Rodríguez-Gil JE, Miró J. 2015. Cryotolerance of stallion spermatozoa is related to ROS production and mitochondrial membrane potential rather than to the integrity of sperm nucleus. Andrology 3(2):395-407.

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Journal of Veterinary Andrology is a peer-reviewed, No-fee Open-access journal.  

Journal of Veterinary Andrology does not charge any publication fees to authors.

The journal allows the author(s) to hold the copyright without restrictions and will retain publishing rights without restrictions


Licencia Creative Commons Atribución 4.0 Internacional.

JOURNAL OF VETERINARY ANDROLOGY esta indexada en/ is indexed in:

Índice y Biblioteca Electrónica de Revistas Venezolanas de Ciencias y Tecnología (REVENCYT). Código RVJ002

International Scientific Indexing (ISI)

Scientific Indexing Services. Journal ID 3410

International Institute of Organized Research (I2OR

Directory of Open Access Scholarly Resources (ROAD)

BASE (Bielefeld Academic Search Engine)

Directory of Research Journals Indexing (DRJI)

Actualidad Iberoamericana

Directory of Open Access Journals (DOAJ)

International Veterinary Information Service (IVIS)

Matriz de Informacion para el Análisis de Revistas (MIAR)

Scientific Journal Impact Factor (SJIF)

CiteFactor Academic Scientific Journals


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 International Veterinary Information Service - IVIS 


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