Hostname: page-component-cd9895bd7-gvvz8 Total loading time: 0 Render date: 2024-12-28T03:50:16.308Z Has data issue: false hasContentIssue false

Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination

Published online by Cambridge University Press:  11 June 2014

A. Johannisson*
Affiliation:
Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Box 7011, Uppsala, Sweden
A. Lundgren
Affiliation:
Department of Clinical Sciences, Division of Reproduction, Swedish University of Agricultural Sciences, Box 7054, Uppsala, Sweden
P. Humblot
Affiliation:
Department of Clinical Sciences, Division of Reproduction, Swedish University of Agricultural Sciences, Box 7054, Uppsala, Sweden
J. M. Morrell
Affiliation:
Department of Clinical Sciences, Division of Reproduction, Swedish University of Agricultural Sciences, Box 7054, Uppsala, Sweden
Get access

Abstract

The decrease in foaling rates after artificial insemination with cooled semen warrants the search for new predictors of fertility. The objectives were to investigate levels of naturally occurring reactive oxygen species (ROS) in cooled, stored stallion semen doses for artificial insemination (AI), and their relationship with parameters of semen quality and with pregnancy rate. Semen was collected from warmblood stallions (n=15) and used to prepare commercial semen doses for AI. Sperm quality was evaluated after cooled transport to the laboratory overnight. The results were correlated with observed foaling and pregnancy rates. Hydroethidine and dichlorodihydrofluorescein diacetate were used as indicators for the ROS superoxide and hydrogen peroxide, respectively. Sperm morphology, motility, plasma membrane integrity and chromatin integrity were also evaluated. These variables were correlated with each other and with pregnancy rates. We found a high inter-individual variation in the ROS levels between stallions. The proportion of live, hydrogen peroxide-negative spermatozoa was correlated with progressive motility, whereas live hydrogen peroxide-negative spermatozoa and chromatin damage were negatively correlated, indicating that low levels of hydrogen peroxide were correlated with good chromatin integrity. The percentage of dead hydrogen peroxide-positive sperm was negatively related to the foaling rate. The negative relationships were stronger when combining results from both assays for ROS. These results for stored semen samples indicate that high individual variation exists for superoxide and hydrogen peroxide measurements, and that ROS status can influence sperm quality. Thus, ROS may be some of the factors influencing fertility. Moreover, combinations of ROS variables improved the correlation with fertility, indicating the usefulness of including these variables in a future model for prediction of the fertility of a semen sample.

Type
Research Article
Copyright
© The Animal Consortium 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aitken, RJ 1995. Free radicals, lipid peroxidation and sperm function. Reproduction, Fertility and Development 7, 659668.CrossRefGoogle ScholarPubMed
Alvarez, JG, Touchstone, JC, Blasco, L and Storey, BT 1987. Spontaneous lipid peroxidation and production of hydrogen peroxide in human spermatozoa. Superoxide dismutase as major enzyme protectant against oxygen toxicity. Journal of Andrology 8, 338348.CrossRefGoogle ScholarPubMed
Amann, RP 2005. Weaknesses in reports of ‘fertility’ for horses and other species. Theriogenology 63, 698715.Google Scholar
Aurich, C 2008. Recent advances in cooled-semen technology. Animal Reproduction Science 107, 268275.Google Scholar
Aurich, JE, Schonherr, U, Hoppe, H and Aurich, C 1997. Effects of antioxidants on motility and membrane integrity of chilled-stored stallion semen. Theriogenology 48, 185192.Google Scholar
Batellier, F, Duchamp, G, Vidament, M, Arnaud, G, Palmer, E and Magistrini, M 1998. Delayed insemination is successful with a new extender for storing fresh equine semen at 15C under anaerobic conditions. Theriogenology 50, 229236.Google Scholar
Baumber, J, Ball, BA, Gravance, CG, Medina, V and 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, 895902.CrossRefGoogle ScholarPubMed
Brinkerhoff, JM, Love, CC, Thompson, JA, Blodgett, G, Teague, SR and Varner, DD 2010. Influence of mare age, pre-breeding mare status, breeding method, and stallion on first-cycle pregnancy rates on a large commercial breeding farm. Animal Reproduction Science 121S, 159160.Google Scholar
Colenbrander, B, Gadella, BM and Stout, TAE 2003. The predictive value of semen analysis in the evaluation of stallion fertility. Reproduction in Domestic Animals 38, 305311.Google Scholar
Ermilov, A, Diamond, MP, Sacco, AG and Dozortsev, DD 1999. Culture media and their components differ in their ability to scavenge reactive oxygen species in the plasmid relaxation assay. Fertility and Sterility 72, 154157.Google Scholar
Evenson, DP, Larsson, KL and Jost, LK 2002. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. Journal of Andrology 23, 2543.Google Scholar
Floreani, M, Napoli, E and Palatini, P 2002. Role of antioxidant defences in the species-specific response of isolated atria to menadione. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 132, 143151.Google Scholar
Griveau, JF, Dumont, E, Renard, P, Callegari, JP and LeLannou, D 1995. Reactive oxygen species, lipid peroxidation and enzymatic defence systems in human spermatozoa. Journal of Reproduction and Fertility 103, 1726.Google Scholar
Guthrie, HD and Welch, GR 2006. Determination of intracellular reactive species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry. Journal of Animal Science 84, 20892100.Google Scholar
Johannisson, A, Morrell, JM, Thorén, J, Jönsson, M, Dalin, A-M and Rodriguez-Martinez, H 2009. Colloidal centrifugation with Androcoll-E™ prolongs stallion sperm motility, viability and chromatin integrity. Animal Reproduction Science 116, 119128.Google Scholar
Katila, T, Reilas, T, Nivola, K, Peltonen, T and Virtala, AM 2010. A 15-year survey of reproductive efficiency of standardbred and finnhorse trotters in Finland – descriptive results. Acta Veterinaria Scandinavica 52, 40.CrossRefGoogle ScholarPubMed
Leclerc, P, de Lamirande, E and Gagnon, C 1997. Interaction between Ca2+, cyclic 3`, 5`-adenosine monophosphate, the superoxide anion, and tyrosine phosphorylation pathways in the regulation of human spermatozoa. Journal of Andrology 19, 434443.CrossRefGoogle Scholar
Morrell, JM, Johannisson, A, Dalin, A-M, Hammar, L, Sandebert, T and Rodriguez-Martinez, H 2008. Sperm morphology and chromatin integrity in Swedish warmblood stallions and their relationship to pregnancy rate. Acta Veterinaria Scandinavica 50, 2.Google Scholar
Morrell, JM, Winblad, C, Georgakas, A, Stuhtmann, G, Humblot, P and Johannisson, A 2013. Reactive oxygen species in stallion semen can be affected by season and colloid centrifugation. Animal Reproduction Science 140, 6269.Google Scholar
Ortega Ferrusola, C, Gonzáles Fernández, L, Morrell, JM, Salazar Sandoval, C, Macías García, B, Rodríguez-Martinez, H, Tapia, JA and Peña, FJ 2009. Lipid peroxidation, assessed with BODIPY-C11, increases after cryopreservation of stallion spermatozoa, is stallion dependent and is related to apoptotic-like changes. Reproduction 138, 5563.CrossRefGoogle ScholarPubMed
Rota, A, Furzi, C, Panzini, D and Camillo, F 2004. Studies on motility and fertility of cooled stallion spermatozoa. Reproduction in Domestic Animals 39, 103109.Google Scholar
van Wienen, M, Johannisson, A, Wallgren, M, Parlevliet, J and Morrell, JM 2011. Single layer centrifugation with Androcoll™-P can be scaled-up to process larger volumes of boar semen. ISRN Veterinary Science 2011, 18. Article ID: 548385.Google Scholar
Zielonka, J, Hardy, M and Kalyanaranman, B 2009. HPLC study of oxidation products of hydroethidine in chemical and biological systems: ramifications in superoxide measurements. Free Radical Biology and Medicine 46, 329338.CrossRefGoogle ScholarPubMed