ARSci  Vol.3 No.3 , August 2015
Azteca Breed Horse Epididymal Sperm Evaluation: A Comparison of Head, Corpus and Cauda Sperm Quality
Abstract: The main objective of this study was to determine differences in sperm recovered from the three segments of the epididymis, i.e., head, corpus and cauda, using routine measurements of sperm quality and DNA fragmentation assessment. There was a larger sperm concentration (P < 0.05) in the head (3.34 × 106 sperm/mL) compared with the corpus and cauda. The percentage of normal morphology was higher in the cauda but had lower motility (P < 0.05). The DNA fragmentation index was higher for sperm obtained from the head and body of the epididymis (P < 0.01) compared with the cauda. The DNA fragmentation index values were 12% higher (P < 0.01) using Wright stain compared with DAPI stain in sperm from the head and corpus of the epididymis, but similar in the cauda. Spermatobioscopy parameters match the maturation changes that occur in the epididymal tract with higher quality sperm being from the cauda, but with a lower concentration compared to sperm found in the head.
Cite this paper: Parra-Forero, L. , Mendoza, G. , Góngora, A. , López Fernández, M. , Cruz, L. , Montiel, A. , Kjelland, M. and García-Contreras, A. (2015) Azteca Breed Horse Epididymal Sperm Evaluation: A Comparison of Head, Corpus and Cauda Sperm Quality. Advances in Reproductive Sciences, 3, 57-65. doi: 10.4236/arsci.2015.33007.

[1]   Braun, J., Sakai, M., Hochi, S. and Oguri, N. (1994) Preservation of Ejaculated and Epididymal Stallion Spermatozoa by Cooling and Freezing. Theriogenology, 41, 809-818.

[2]   Bruemmer, J.E. (2006) Collection and Freezing of Epididymal Stallion Sperm. Veterinary Clinics of North America: Equine Practice, 22, 677-682.

[3]   Guimarães, T., Lopes, G., Ferreira, P., Leal, I. and Rocha, A. (2012) Characteristics of Stallion Epididymal Spermatozoa at Collection and Effect of Two Refrigeration Protocols on the Quality of the Frozen/Thawed Sperm Cells. Animal Reproduction Science, 136, 85-89.

[4]   Samper, J.C. (2009) Equine Breeding Management and Artificial Insemination. Elsevier Health Sciences.

[5]   Ball, B.A. (2008) Diagnostic Methods for Evaluation of Stallion Subfertility: A Review. Journal of Equine Veterinary Science, 28, 650-665.

[6]   Betancur, G.R., Suárez, A.ú. and Rojano, B.A. (2014) Técnicas para el análisis de la fertilidad potencial del semen equino. (Techniques for Analyzing the Potential Fertility of Stallion Semen) (Métodos de avaliação do potencial de fertilidade do sêmen equino). Revista CES Medicina Veterinaria y Zootecnia, 8, 115-127.

[7]   Papa, F.O., Melo, C. and Fioratti, E. (2008) Freezing of Stallion Epididymal Sperm. Animal Reproduction Science, 107, 293-301.

[8]   Monteiro, G.A., Papa, F.O., Zahn, F.S., Dellaqua Jr., J.A., Melo, C.M., Maziero, R.R.D., Avanzi, B.R., Alvarenga, M.A. and Guasti, P.N. (2011) Cryopreservation and Fertility of Ejaculated and Epididymal Stallion Sperm. Animal Re- production Science, 127, 197-201.

[9]   Heise, A., Thompson, P.N. and Gerber, D. (2011) Influence of Seminal Plasma on Fresh and Post-Thaw Parameters of Stallion Epididymal Spermatozoa. Animal Reproduction Science, 123, 192-201.

[10]   Dias, G., Retamal, C., Tobella, L., Arnholdt, A. and Lopez, M. (2006) Nuclear Status of Immature and Mature Stallion Spermatozoa. Theriogenology, 66, 354-365.

[11]   Clemmons, A.J., Thompson, D. and Johnson, L. (1995) Local Initiation of Spermatogenesis in the Horse. Biology of Reproduction, 52, 1258-1267.

[12]   Baumann, C., Daly, C.M., McDonnell, S.M., Viveiros, M.M. and De La Fuente, R. (2011) Chromatin Configuration and Epigenetic Landscape at the Sex Chromosome Bivalent during Equine Spermatogenesis. Chromosoma, 120, 227- 244.

[13]   Varner, D.D., Blanchard, T.L., Brinsko, S.P., Love, C.C., Taylor, T.S. and Johnson, L. (2000) Techniques for Evaluating Selected Reproductive Disorders of Stallions. Animal Reproduction Science, 60-61, 493-509.

[14]   Morte, M.I., Rodrigues, A.M., Soares, D., Rodrigues, A.S., Gamboa, S. and Ramalho-Santos, J. (2008) The Quantification of Lipid and Protein Oxidation in Stallion Spermatozoa and Seminal Plasma: Seasonal Distinctions and Correlations with DNA Strand Breaks, Classical Seminal Parameters and Stallion Fertility. Animal Reproduction Science, 106, 36-47.

[15]   Toro, E., Fernández, S., Colomar, A., Casanovas, A., álvarez, J.G., López-Teijón, M. and Velilla, E. (2009) Processing of Semen Can Result in Increased Sperm DNA Fragmentation. Fertility and Sterility, 92, 2109-2112.

[16]   Garcia-Macias, V., Martinez-Pastor, F., Alvarez, M., Garde, J.J., Anel, E., Anel, L. and de Paz, P. (2006) Assessment of Chromatin Status (SCSA®) in Epididymal and Ejaculated Sperm in Iberian Red Deer, Ram and Domestic Dog. Theriogenology, 66, 1921-1930.

[17]   Muradas, P.R., Weiss, R.R., Kozicki, L.E., Granemann, L.C., Moreira, N., Souza, J., Santos, I.W., Abreu, R.A., Green, K. and Milleo, T.C. (2013) The Viability of Cryopreserved Spermatozoa Recovered from the Cauda Epididymis of Equines and the Time of Collection Post-Orchiectomy. Asian Journal of Agriculture and Food Science.

[18]   Committee, F.o.A.S.S.W. (2010) Federation of Animal Science Societies, Champaign-Urbana.

[19]   Bedford, S., Varner, D. and Meyers, S. (1999) Effects of Cryopreservation on the Acrosomal Status of Stallion Spermatozoa. Journal of Reproduction and Fertility. Supplement, No. 56, 133-140.

[20]   Meyers, S. (2001) Equine Sperm-Oocyte Interaction: The Role of Sperm Surface Hyaluronidase. Animal Reproduction Science, 68, 291-303.

[21]   Farlin, M., Jasko, D., Graham, J. and Squires, E. (1992) Assessment of Pisum sativum Agglutinin in Identifying Acrosomal Damage in Stallion Spermatozoa. Molecular Reproduction and Development, 32, 23-27.

[22]   López-Fernández, C., Crespo, F., Arroyo, F., Fernández, J.L., Arana, P., Johnston, S.D. and Gosálvez, J. (2007) Dynamics of Sperm DNA Fragmentation in Domestic Animals: II. The Stallion. Theriogenology, 68, 1240-1250.

[23]   Fernández, J., Gosálvez, J., Santiso, R., Goyanes, V. and López-Fernández, C. (2008) Adaptation of the Sperm Chromatin Dispersion (SCD) Test to Determine DNA Fragmentation from Bull Sperm. New Cell Apoptosis Research. Nova Science Publishers, Inc., New York.

[24]   González-Marín, C., Roy, R., López-Fernández, C., Diez, B., Carabaño, M., Fernández, J., Kjelland, M., Moreno, J. and Gosálvez, J. (2011) Bacteria in Bovine Semen Can Increase Sperm DNA Fragmentation Rates: A Kinetic Experimental Approach. Animal Reproduction Science, 123, 139-148.

[25]   Fernández, J.L., Muriel, L., Goyanes, V., Segrelles, E., Gosálvez, J., Enciso, M., LaFromboise, M. and De Jonge, C. (2005) Simple Determination of Human Sperm DNA Fragmentation with an Improved Sperm Chromatin Dispersion test. Fertility and Sterility, 84, 833-842.

[26]   Zee, Y.P., López-Fernández, C., Arroyo, F., Johnston, S.D., Holt, W.V. and Gosalvez, J. (2009) Evidence that Single-Stranded DNA Breaks Are a Normal Feature of Koala Sperm Chromatin, While Double-Stranded DNA Breaks Are Indicative of DNA Damage. Reproduction, 138, 267-278.

[27]   Dogan, S., Vargovic, P., Oliveira, R., Belser, L.E., Kaya, A., Moura, A., Sutovsky, P., Parrish, J., Topper, E. and Memili, E. (2015) Sperm Protamine-Status Correlates to the Fertility of Breeding Bulls. Biology of Reproduction, 92, 92.

[28]   Kjelland, M., González-Marín, C., Gosálvez, J., López-Fernández, C., Lenz, R., Evans, K. and Moreno, J. (2011) DNA Fragmentation Kinetics and Postthaw Motility of Flow Cytometric-Sorted White-Tailed Deer Sperm. Journal of Animal Science, 89, 3996-4006.

[29]   Green, S.B. and Salkind, N.J. (2010) Using SPSS for Windows and Macintosh: Analyzing and Understanding Data. Prentice Hall Press, Upper Saddle River.

[30]   Pallant, J. (2010) SPSS Survival Manual: A Step by Step Guide to Data Analysis Using SPSS. McGraw-Hill International, New York.

[31]   Vieira, L., Gadea, J., García-Vázquez, F., Avilés-López, K. and Matás, C. (2013) Equine Spermatozoa Stored in the Epididymis for up to 96 h at 4 °C Can Be Successfully Cryopreserved and Maintain Their Fertilization Capacity. Animal Reproduction Science, 136, 280-288.

[32]   Neto, C.R., Monteiro, G.A., Soares, R.F., Pedrazzi, C., Dell’aqua Jr., J.A., Papa, F.O. and Alvarenga, M.A. (2013) Effect of Removing Seminal Plasma Using a Sperm Filter on the Viability of Refrigerated Stallion Semen. Journal of Equine Veterinary Science, 33, 40-43.

[33]   Monteiro, G.A., Guasti, P.N., Rocha, A.S., Martin, I., Sancler-Silva, Y.F.R., Freitas Dell’Aqua, C.P., Dell’Aqua Jr., J.A. and Papa, F.O. (2013) Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm. Journal of Equine Veterinary Science, 33, 169-173.

[34]   Fouchécourt, S., Métayer, S., Locatelli, A., Dacheux, F. and Dacheux, J.-L. (2000) Stallion Epididymal Fluid Proteome: Qualitative and Quantitative Characterization; Secretion and Dynamic Changes of Major Proteins. Biology of Reproduction, 62, 1790-1803.

[35]   James, A., Green, H., Hoffman, S., Landry, A., Paccamonti, D. and Godke, R. (2002) Preservation of Equine Sperm Stored in the Epididymis at 4 °C for 24, 48, 72 and 96 Hours. Theriogenology, 58, 401-404.

[36]   Monteiro, G.A., Freitas-Dell’Aqua, C.P., Guasti, P.N., Dell’Aqua Jr., J.A., Alvarenga, M.A., Landim, F.C. and Papa, F.O. (2013) Comparison of Apoptotic Cells between Cryopreserved Ejaculated Sperm and Epididymal Sperm in Stallions. Journal of Equine Veterinary Science, 33, 552-556.

[37]   Leme, D.P., Papa, F.O. and Roser, J.F. (2012) Reproductive Characteristics of Stallions during the Breeding and Non-Breeding Season in a Tropical Region. Tropical Animal Health and Production, 44, 1703-1707.

[38]   Claes, A., Ball, B.A., Almeida, J., Corbin, C.J. and Conley, A.J. (2013) Serum Anti-Müllerian Hormone Concentrations in Stallions: Developmental Changes, Seasonal Variation, and Differences between Intact Stallions, Cryptorchid Stallions, and Geldings. Theriogenology, 79, 1229-1235.

[39]   Cary, J.A., Madill, S., Farnsworth, K., Hayna, J.T., Duoos, L. and Fahning, M.L. (2004) A Comparison of Electroejaculation and Epididymal Sperm Collection Techniques in Stallions. The Canadian Veterinary Journal, 45, 35.

[40]   Olaciregui, M., Gil, L., Monton, A., Luno, V., Jerez, R.A. and Marti, J.I. (2014) Cryopreservation of Epididymal Stallion Sperm. Cryobiology, 68, 91-95.

[41]   Johnson, L. (1995) Efficiency of Spermatogenesis. Microscopy Research and Technique, 32, 385-422.

[42]   Parks, J., Lee, D., Huang, S. and Kaproth, M. (2003) Prospects for Spermatogenesis in Vitro. Theriogenology, 59, 73- 86.

[43]   Hess, R.A. and de Franca, L.R. (2009) Spermatogenesis and Cycle of the Seminiferous Epithelium. In Molecular Mechanisms in Spermatogenesis (pp. 1-15). Springer, New York.

[44]   Dacheux, J.-L., Castella, S., Gatti, J.L. and Dacheux, F. (2005) Epididymal Cell Secretory Activities and the Role of Proteins in Boar Sperm Maturation. Theriogenology, 63, 319-341.

[45]   Gatti, J.-L., Castella, S., Dacheux, F., Ecroyd, H., Metayer, S., Thimon, V. and Dacheux, J.-L. (2004) Post-Testicular Sperm Environment and Fertility. Animal Reproduction Science, 82, 321-339.

[46]   Dacheux, J., Belghazi, M., Lanson, Y. and Dacheux, F. (2006) Human Epididymal Secretome and Proteome. Molecular and Cellular Endocrinology, 250, 36-42.

[47]   Sostaric, E., Aalberts, M., Gadella, B. and Stout, T.A. (2008) The Roles of the Epididymis and Prostasomes in the Attainment of Fertilizing Capacity by Stallion Sperm. Animal Reproduction Science, 107, 237-248.

[48]   Morris, L., Tiplady, C. and Allen, W. (2002) The in Vivo Fertility of Cauda Epididymal Spermatozoa in the Horse. Theriogenology, 58, 643-646.

[49]   Heise, A., Kähn, W., Volkmann, D.H., Thompson, P.N. and Gerber, D. (2010) Influence of Seminal Plasma on Fertility of Fresh and Frozen-Thawed Stallion Epididymal Spermatozoa. Animal Reproduction Science, 118, 48-53.

[50]   Honaramooz, A., Snedaker, A., Boiani, M., Schöler, H., Dobrinski, I. and Schlatt, S. (2002) Sperm from Neonatal Mammalian Testes Grafted in Mice. Nature, 418, 778-781.

[51]   Sato, T., Katagiri, K., Gohbara, A., Inoue, K., Ogonuki, N., Ogura, A., Kubota, Y. and Ogawa, T. (2011) In Vitro Production of Functional Sperm in Cultured Neonatal Mouse Testes. Nature, 471, 504-507.

[52]   Enciso, M., López-Fernández, C., Fernández, J.L., García, P., Gosálbez, A. and Gosálvez, J. (2006) A New Method to Analyze Boar Sperm DNA Fragmentation under Bright-Field or Fluorescence Microscopy. Theriogenology, 65, 308- 316.

[53]   Sun, J.-G., Jurisicova, A. and Casper, R.F. (1997) Detection of Deoxyribonucleic Acid Fragmentation in Human Sperm: Correlation with Fertilization in Vitro. Biology of Reproduction, 56, 602-607.

[54]   Chohan, K.R., Griffin, J.T., Lafromboise, M., Jonge, C.J. and Carrell, D.T. (2006) Comparison of Chromatin Assays for DNA Fragmentation Evaluation in Human Sperm. Journal of Andrology, 27, 53-59.