This study was aimed to analyze the metabolic activity and membrane integrity of boar spermatozoa following storage in long-term semen extenders. Boar semen was diluted with AndrohepR EnduraGuardTM (AeG), DILU-Cell (DC), SafeCell PlusTM (SCP) and Vitasem LD (VLD) extenders and stored for 10 days at 17oC. Parameters of the analyzed sperm metabolic activity included total motility (TMOT), progressive motility (PMOT), high mitochondrial membrane potential (MMP) and ATP content, whereas those of the membrane integrity included plasma membrane integrity (PMI) and normal apical ridge (NAR) acrosome. Extender type was a significant (P < 0.05) source of variation in all the analyzed sperm parameters, except for ATP content. Furthermore, the storage time had a significant effect (P < 0.05) on the sperm metabolic activity and membrane integrity during semen storage. In all extenders the metabolic activity and membrane integrity of the stored spermatozoa decreased continuously over time. Among the four analyzed extenders, AeG and SCP showed the best performance in terms of TMOT and PMI on Days 5, 7 and 10 of storage. Marked differences in the proportions of spermatozoa with high MMP were observed between the extenders, particularly on Day 10 of storage. There were not any marked differences in sperm ATP content between the extenders, regardless of the storage time. Furthermore, the percentage of spermatozoa with NAR acrosomes decreased during prolonged storage, being markedly lower in DC-diluted semen compared with semen diluted with either AeG or SCP extender. The results of this study indicated that components of the long-term extenders have different effects on the sperm functionality and prolonged semen longevity by delaying the processes associated with sperm ageing during liquid storage.
Anil SS, Larriestra A, Deen J, Morrison RB, Minion L (2004) A retrospective study on the preserving capacity of a commercial boar semen extender. Theriogenology 62: 425-436.
De Ambrogi M, Ballester J, Saravia F, Caballero I, Johannisson A, Wallgren M, Andersson M, Rodriquez-Martinez H (2006) Effect of storage in short- and long-term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa. Int J Androl 29: 543-552.
Dziekońska A, Fraser L, Strzeżek J (2009) Effects of different storage temperatures on the metabolic activity of spermatozoa following liquid storage of boar semen. J Anim Feed Sci 18: 638-649.
Dziekońska A, Strzeżek J (2011) Boar variability affects sperm metabolism activity in liquid stored semen at 5oC. Pol J Vet Sci 14: 21-27.
Fantinati P, Zannoni A, Bernardini C, Forni M, Tattini A, Seren E, Bacci ML (2009) Evaluation of swine fertilisation medium (SFM) efficiency in preserving spermatozoa quality during long-term storage in comparison to four commercial swine extenders. Animal 3: 269-274.
Flesch FM, Gadella BM (2000) Dynamics of the mammalian sperm plasma membrane in the process of fertilization. Biochim Biophys Acta 1469: 197-235.
Fraser L, Dziekońska A, Strzeżek R, Strzeżek J (2007) Dialysis of boar semen prior to freezing-thawing: its effects on post-thaw sperm characteristics. Theriogenology 67: 994-1003.
Fraser L, Gorszczaruk K, Lecewicz M, Strzeżek J (2003) Age-related changes and seasonal variation in boar sperm metabolism during liquid storage at 5o and 16oC. J Anim Feed Sci 12: 803-811.
Gadea J (2003) Review: Semen extenders used in the artificial insemination of swine. Spanish J Agric Res 1(2): 17-27.
Garner DL, Johnson LA (1995) Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod 53: 276-284.
Gączarzewicz D, Piasecka M, Udała J, Błaszczyk B, Stankiewicz T, Laszczyńska M (2010) Plasma membrane changes during the liquid storage of boar spermatozoa: a comparison of methods. Acta Vet Hung 58: 105-116.
Gerrits RJ, Lunney JK, Johnson LA, Pursel VG, Kraeling RR, Rohrer GA, Dobrinsky JR (2005) Perspectives for artificial insemination and genomics to improve global swine populations. Theriogenology 63: 283-299.
Gogol P, Szczęśniak-Fabiańczyk B, Wierzchoś-Hilczer A (2009) The photon emission, ATP level and motility of boar spermatozoa during liquid storage. Reprod Biol 9: 39-49.
Guthrie HD, Welch GR, Long JA (2008) Mitochondrial function and reactive oxygen species action in relation to boar motility. Theriogenology 70: 1209-1215.
Huo LJ, Ma XH, Yang ZM (2002) Assessment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage. Theriogenology 58: 1349-1360.
Johnson LA, Weitze KF, Fiser P, Maxwell WM (2000) Storage of boar semen. Anim Reprod Sci 62: 143-172.
Kaeoket K, Srisowanna T, Wichaidit U, Chanapiwat P, Manee-in S (2010) Comparative study on six different long term commercial extenders for fresh boar semen. Thai J Vet Med 40: 257-263.
Kommisrud E, Paulenz H, Sehested E, Grevle IS (2002) Influence of boar and semen parameters on motility and acrosome integrity in liquid boar semen stored for five days. Acta Vet Scand 43: 49-55.
Kumaresan A, Kadirvel G, Bujarbaruah KM, Bardoloi RK, Das A, Kumar S, Naskar S (2008) Preservation of boar semen at 18oC induces lipid peroxidation and apoptosis like changes in spermatozoa. Anim Reprod Sci 110: 162-171.
Kuster CE, Althouse GC (1999) The fecundity of porcine semen stored for 2 to 6 days in Androhep and X-cell extenders. Theriogenolgy 52: 365-376.
Medrano A, Pen˜a A, Rigau T, Rodrı´guez-Gil JE (2005) Variations in the proportion of glycolytic/non-glycolytic energy substrates modulate sperm membrane integrity and function in diluted boar samples stored at 15-17oC. Reprod Domest Anim 40: 448-453.
Rodr´ıguez-Mart´ınez H (2003) Laboratory semen assessment and prediction of fertility: still utopia? Reprod Domest Anim 38: 312-318.
Storey BT (2008) Mammalian sperm metabolism: oxygen and sugar, friend and foe. Int J Dev Biol 52: 427-437.
Thomas CA, Garner DL, De Jarnette JM, Marshall CE (1998) Effect of cryopreservation on bovine sperm organelle function and viability as determined by flow cytometry. Biol Reprod 58: 786-793.
Trzcińska M, Bryła M, Smorąg Z (2008) Effect of liquid storage on membrane integrity and mitochondrial activity: a new diagnostic method of evaluating boar sperm quality. J Anim Feed Sci 17: 372-380.
Vyt P, Maes D, Dejonckheere E, Castryck F, Van Soom A (2004) Comparative study on five different commercial extenders for boar semen. Reprod Domest Anim 39: 8-12.
Waberski D, Henning H, Petrunkina AM (2011) Assessment of storage effects in liquid preserved boar semen. Reprod Domest Anim 46: 45-48.
Waberski D, Weitze KF, Rath D, Sallman HP (1989) Effect of bovine serum albumin and zwitterionic buffers on stored liquid boar semen. Zuchthygiene 24: 128-133.
Watson PF (1975) Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. Vet Rec 97: 12-15.
World Health Organization (1999) WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction, 4th ed., Cambridge University Press, Cambridge, pp 90-93.