Effect of different extenders on recovery and storage of epididymal ram spermatozoa

Document Type : Research Articles

Authors

1 Unversity of Guilan

2 University of Guilan

Abstract

This experiment was conducted to determine effect of different extender on recovery and storage of epididymal ram spermatozoa. Testes were obtained from eight mature rams which were slaughtered at four different occasions, a pair in each session. Caudal epididymis was isolated and divided to four equal parts. Spermatozoa recovery from each part of the caudal epididymis was performed by milk (M), Tris (T), Tris plus caproic acid (T-C) and sodium citrate (S) extenders. Spermatozoa in each extender were divided into four parts and then the amount of 0 (E0), 5 (E5), 10 (E10) and 15% (E15) egg yolk were added. Samples were chilled to 4 °C and stored for 7 days. The evaluation of sperm motility, viability, membrane integrity and acrosome integrity were performed daily. The results showed that there was interaction between egg yolk and storage time and also egg yolk and extender on sperm motility (p < 0.05). Sperm motility was highest at 15 % egg
yolk (25.6 %) on 7 d (p < 0.05). Sperm motility was higher in ME15 (57.8 %) and T-CE15 (62.3 %) than TE15 (53.4 %) and SE15 (41.9 %; p < 0.05). There was interaction among egg yolk, extender and storage time on acrosome integrity, membrane integrity and sperm viability (p < 0.05). On 7 d, there were no differences among ME15, T-CE15 and TE15 on sperm viability and membrane integrity. Acrosome integrity was higher in T-CE15 (75.25 %) than ME15 (62.5 %) on 7 d (p < 0.05). Therefore, Tris caproic extender was superior to other extender for recovery and storage of ram epididymal spermatozoa.

Keywords


Batellier, F; Magistrini, M; Fauquant, J and Palmer, E (1997) Effect of milk fractions on survival of equine spermatozoa. Theriogenology 48, 391–410.
Cervato, G; Cazzola, R and Cestaro, B (1999) Studies on the antioxidant activity of milk caseins. International Journal of Food Sciences and Nutrition 50, 291–296.
Choong, CH and Wales, RG (1962) The effect of cold shock on spermatozoa. Australian Journal of Biological Sciences 15, 543-551.
Coronel, C; Burgos, C; Gerez, DC; Burgos, NM; Rovai, LE and Blanco, A (1983) Catalytic properties of the sperm-specific lactate dehydrogenase (LDH-X or C4) from different species. Journal of Experimental Zoology 225, 379–385.
De Leeuw, FE; Colenbrander, B and Verkleij, AJ (1991) The role membrane damage plays in cold shock and freezing injury. Reproduction in Domestic Animals 1, 95-104.
Dong, Q; Rodenburg, SE; Huang, C and Vandevoort, CA (2008) Cryopreservation of rhesus monkey (Macaca mulatta) epididymal spermatozoa before and after refrigerated storage. Journal of Andrology 29, 283-292.
Evans, G and Maxwell, WMC (1987). Handling and examination of semen & Salamon’s Artificial Insemination of Sheep and Goats, Evans, G and Maxwell, WMC (eds.), Butterworths, Boston, pp. 97–98.
Gordo, AC; Rodrigues, P; Kurokawa, M; Jellerette, T; Exley, GE; Warner, C and Fissore, R (2002) Intracellular calcium oscillations signal apoptosis rather than activation in vitro aged mouse eggs. Biology of Reproduction 66, 1828–1837.
Guasti, PN; Monteiro, GA; Maziero, RR; Martin, I; Avanzi, BR; Dellaqua, Jr JA and Papa, FO (2013) Effects of pentoxifylline on equine epididymal sperm. Journal of Equine Veterinary Science 33, 1153–1156.
Jeyendran, RS; Van-der-Ven, HH and Zaneveld LJ (1992) The hypoosmotic swelling test: an update. Archives of Andrology 29,105–116.
Kaabia, M; Pazb, P; Alvareza, M; Anela, E; Boixoa, JC; Rouissic, H; Herraezb, P and Anel, L (2003) Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology 60, 1249–1259.
Kitts, DD (2005). Antioxidant properties of casein-phosphopeptides. Trends in Food Science & Technology 16, 549–554.
Kumar, D and Naqvi, SMK (2014) Effect of time and depth of insemination on fertility of Bharat Merino sheep inseminated trans-cervical with frozen-thawed semen. Journal of Animal Science and Technology 56, 1-8.
Leboeuf, B; Guillouet, P; Batellier, F; Bernelas, D; Bonne, JL; Forgerit, Y; Renaud, G and Magistrini, M (2003) Effect of native phosphocaseinate on the in vitro preservation of fresh semen. Theriogenology 60, 867-877.
Lone, FA; Islam, R; Khan, MZ and Sofi, KA (2011) Effect of transportation temperature on the quality of cauda epididymal spermatozoa of ram. Animal Reproduction Science 123, 54–59.
Lopez-Saez, A1; Ortiz, N; Gallego, L and Garde, JJ (2000)Liquids storage (5 degrees C) of ram semen in different diluents. Archives of Andrology 44, 155-164.
Martin, G; Sabido, O; Durand, P and Levy, R (2005) Phosphatidylserine externalization in human sperm induced by calcium ionophore A23187: relationship with apoptosis, membrane scrambling and the acrosome reaction. Human Reproduction 20, 3459–3468.
Morris, L; Tiplady, C and Allen, WR (2002) The in vivo fertility of caudal epididymal spermatozoa in horse. Theriogenology 58, 643-646.
O’Hara, L; Hanrahan, JP; Richardson, L; Donovan, A; Fair, S; Evans, ACO and Lonergan, P (2010) Effect of storage duration, storage temperature, and diluent on the viability and fertility of fresh ram sperm. Theriogenology 73,541–549.
O’Shea, T and Wales, RG (1966) Effect of casein, lecithin, glycerol, and storage at 5°C on diluted ram and bull semen. Australian Journal of Biological Sciences 19, 871-882.
Paulenz, H; Söderquist, L; Perez-Pe, R and Berg, KA (2002) Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology 57, 823–836.
Rakha, BA; Hussain, I; Akhter, S; Ullah, N; Andrabi, SM and Ansari, MS (2013) Evaluation of Tris-citric
acid, skim milk and sodium citrate extenders for liquid storage of Punjab Urial (Ovis vignei punjabiensis) spermatozoa. Reproductive Biology 13, 238-42.
Roostaei-Ali Mehr, M; Chambary, B and Ghavi Hossein-Zadeh, N (2013) Effect of different diluents and storage time on field fertility of cooled ram semen after vaginal insemination. Small Ruminant Research 115, 82– 85.
Roostaei-Ali Mehr, M; Mousavi, M; and Ghadamyari, M (2015) Effect of seminal plasma proteins on membrane cholesterol efflux of ram epididymal spermatozoa. Small Ruminants Research 129, 88–91.
Santiago-Moreno, J; Toledano-Diaz, A; Pulido- Pastor, A; Dorado, J; Gomez-Brunet, A and Lopez-Sebastian, A (2006) Effect of egg yolk concentration on cryopreserving Spanish ibex (Capra pyrenaica) epididymal spermatozoa. Theriogenology 66, 1219-1226.
SAS (2002) SAS Users Guide: Statistics, Version 9.1 Edition, SAS; Inc., Cary, NC, USA.
Simpson, AM; Swan, MA and White, IG (1987) Calcium uptake, respiration, and ultrastructure of sperm exposed to ionophore A23187. Archives of Andrology 19, 5–18.
Varisli, O; Uguz, C; Agca, C and Agca,
Y (2009) Effect of chilling on the motility and acrosomal integrity of rat sperm in the presence of various extenders. Journal of the American Association for Laboratory Animal Science 48, 499-505.
Varisli, O; Uguz, C; Agca, C and Agca, Y (2009) Motility and acrosomal integrity comparisons between electro-ejaculated and epididymal ram sperm after exposure to a range of anisosmotic solutions, cryoprotective agents and low temperatures. Animal Reproduction Sciences 110, 256-68.
Vijayaraghavan, S; Bhattacharyya, A and Hoskins, DD (1989) Calcium uptake by bovine epididymal spermatozoa is regulated by the redox state of the mitochondrial pyridine nucleotides. Biology of Reproduction 40, 744–751.
Watson, PF and Martin, IC (1975) Influence of some fractions of egg yolk on the survival of ram spermatozoa at 5ºC. Australian Journal of Biological Sciences 28,145-152.
Wierzbowski, S and Kareta, W (1993) An assessment of sperm motility estimation for evaluation in rams. Theriogenology 40, 205–209.
Witte, TS and Schäfer-Somi S (2007) Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa. Animal Reproduction Science 102, 181–193.