Influences of Monosaccharides and Disaccharides Supplementations in Tris Media on the Motility Patterns of Fresh and Chilled Small Ruminant Spermatozoa

Document Type : Research Article


Division of Animal Production, Department of Agriculture, Atomic Energy Commission, Damascus, Syria.


In this study, the effects of monosaccharides, including glucose and fructose, and disaccharides, namely sucrose and trehalose, in eight Tris media on the motility patterns of small ruminants spermatozoa were investigated. Fresh and chilled semen samples from five Awassi rams and five Shami bucks were diluted in TBM and TEY containing 50 mM of the four different sugar types. The characteristics of spermatozoa motility were analyzed using a computer-assisted sperm analyzer (CASA). Fresh ram spermatozoa incubated in a TBM-fructose medium had the highest CASA values with no differences between the motility values generated from the fructose- and glucose-supplemented media. Trehalose reduced the values of velocity parameters, including VAP, VCL, and VSL for fresh ram sperm. Sucrose was the most influential sugar in raising the values of motility parameters MOT%, PMOT%, VAP, VCL, and VSL for fresh bucks spermatozoa, while trehalose generally had an important positive effect on chilled buck sperms. No significant differences (p > 0.05) were recorded for sperm trajectory parameters where the values of STR% and LIN% for the two ruminant species and the two spermatozoa types did not significantly differ between the eight media. It was concluded that during the first hours of in vitro incubation and based on the incubation temperature, the velocity parameters of small ruminant spermatozoa were the most affected CASA characteristics by monosaccharides and disaccharides supplementations in Tris semen media.


Main Subjects

1.    Bustani GS, Baiee FH. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Vete World 2021; 14(5): 1220-1233. Doi: 10.14202/vetworld.2021.1220-1233
2.    Rigau T, Rivera M, Palomo MJ, Fernandez-Novell, JM, Mogas T, Ballester J.  Differential effects of glucose and fructose on hexose metabolism in dog spermatozoa. Reproduction 2002; 123: 579–91. Doi: 10.1530/rep.0.1230579
3.    Rodríguez- Gil JE. Mammalian sperm energy resources management and survival during conservation in refrigeration. Reprod  Domest Anim 2006; 41(2): 11-20.  Doi:10.1111/j.1439-0531.2006.00765
4.    Rigau T, Farre M, Ballester J, Mogas T, Peiia A, Rodriguez-Gil JE. Effects of glucose and fructose on motility paterns of dog spermatozoa from fresh ejaculates. Theriogenology 2001: 56: 601-615. Doi: 10.1016/s0093-691x(01)00609-4  
5.    Mann T. Biochemistry of semen. In: Greep RO, Astwood EB (ed): Handbook of Physiology. Washington DC, American Physiology Society, Section, 1975; 67(5): 321- 347.
6.    Nagai T, Yamaguchi K, Moriwaki C. Studies on the effects of sugars on washed human sperm motility. J Pharmacobiodyn, 1982; 5: 564–7. Doi: 10.1248/bpb1978.5.564.
7.    Johnson LA, Welch GR, Rens W. The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI. J Anim Sci 1999; 2: 213-220. Doi: 10.2527/1999.77suppl_2213x
8.    Naing SW, Wahid H, Mohd Azam K, Rosnina Y, Zuki AB, Kazhal S, et al. Effect of sugars on characteristics of Boer goat semen after cryopreservation. Anim Reprod Sci 2010; 122: 23–28. Doi: 10.1016/j.anireprosci.2010.06.006
9.    Williams AC, Ford WC. The Role of Glucose in Supporting Motility and Capacitation in Human Spermatozoa. J Androl 2001; 22(4): 680-695. Doi:10.1002/j.1939-4640.2001.tb02229
10.    Aboagla EME, Terada T. Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol Reprod 2003; 69: 1245 – 1250. Doi:10.1095/biolreprod.103.017889
11.    Aboagla EME, Terada T. Effects of the supplementation of trehalose extender containing egg yolk with sodium dodecyl sulphate on the freezability of goat spermatozoa. Theriogenology 2004; 62: 809 – 818. Doi:10.1016/j.theriogenology.2003.12.003 
12.    Zhao J, Xiao G, Zhu W, Fang D, Li N, Han C, et al. Trehalose addition to a Tris-fructose egg yolk extender on quality of ram sperm preserved at 0 ̊C. Rev Bras Zootec 2020; 49: e20200061. Doi:10.37496/rbz4920200061.
13.    Athurupana R, Takahashi D, Ioki S, Funahashi H. Trehalose in glycerol-free freezing extender enhances post-thaw survival of boar spermatozoa. J Reprod Dev 2015; 61:205-210. Doi:10.1262/ jrd.2014-152
14.    Matos-Brito BG, Lima ICS, Pereira JF, Barboza FM, Linard MAB, Aguiar GV, et al. Effect of initial seminal plasma fructose concentration on goat semen storage at 5oC. Arch Zootec 2013; 62; Doi:10.4321/S0004-05922013000100015  
15.    Salamon S, Ritar AJ. Deep freezing of Angora goat semen: Effects of diluent composition and method and rate of dilution on survival of spermatozoa. Aust J  Biol Sci 1982; 35: 295–303.
16.    Purdy PH. A review on goat sperm cryopreservation. Small Rumin Res 2006; 63(3): 215–225. Doi:10.1016/j.smallrumres.2005.02.015
17.    Aisen EG, Medina VH, Venturino A. Cryopreservation and post-thawed fertility of ram semen frozen in different trehalose concentrations. Theriogenology 2002; 57: 1088–1801. Doi: 10.1016/s0093-691x(02)00653-2 
18.    Garde JJ, del Olmo A, Soler AJ, Espeso G, Gomendio M, Roldan ERS. Effect of egg yolk, cryoprotectant, and various sugars on semen cryopreservation in endangered Cuvier’s gazelle (Gazella cuvieri). Anim Reprod Sci 2008;  108(3–4): 384–401.Doi:10.1016/j.anireprosci.2007.09.010
19.    Lapwood KR, Martin ICA. The use of monosaccharides, disaccharides and trisaccharides in synthetic diluents for the storage of ram spermatozoa at 37 ◦C and 5 ◦C. Aust J Biol Sci, 1966; 19: 655–671.
20.    Abdelhakeam AA, Graham EF, Vazquez IA, Chaloner KM. Studies on the absence of glycerol in unfrozen and frozen ram semen:development of an extender for freezing: effects of osmotic pressure, egg yolk levels, type of sugars, and the method of dilution. Cryobiology 1991; 28(1): 43–49. Doi:10.1016/0011-2240(91)90006-A
21.    Alomar M, Zarkawi M, Alzoabi M.A. Analysis of Awassi sperm motility in two media at different levels of temperature, pH and osmolality. Iran J Appl Anim Sci 2018; 8(3): 167-172. 
22.    Ponglowhapan S,  Esseen-Gustavsson B, Forsberg C.L. Influence of glucose and fructose in the extender during long-term storage of chilled canine semen. Theriogenology 2004; 62: 1498–1517.
23.    Curry MR, Kleinhans FW, Watson PF. Measurement of the water permeability of the membranes of boar, ram, and rabbit spermatozoa using concentration-dependent self-quenching of an entrapped fluorophore. Cryobiology 2000; 41: 167–73. Doi:10.1006/cryo.2000.2277
24.    Bedford JM, Hoskins, DD. The mammalian spermatozoon: morphology, biochemistry and physiology. In Marshall’s Physiology of Reproduction 1990; 379–568 Ed. GE Lamming. Churchill Livingstone, Edinburgh.
25.    Jones AR, Connor DE. Fructose metabolism by mature boar spermatozoa. Reprod Fertil Dev 2000; 12: 355–359. Doi:10.1071/RD00116
26.    Rogers BJ, Perreault SD. Importance of glycolysable substrates for in vitro capacitation of human spermatozoa. Biol Reprod 1990; 43: 1064–1069. Doi:10.1095/biolreprod43.6.1064
27.    Garcia MA, Graham EF. Development of a buffer system for dialysis of bovine spermatozoa before freezing. II. Effects of sugars and sugar alcohols on post thaw motility. Theriogenology 1989; 31: 1029–37. Doi:10.1016/0093-691X(89)90486
28.    Molinia FC, Evans G, Maxwell WM. In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa. Reprod Nutr Dev 1994; 34: 491–500. Doi: 10.1051/rnd:19940510 
29.    Pellicer-Rubio MT, Magallon T, Combarnous Y. Deterioration of goat sperm viability in milk extenders is due to a bulbourethral 60-kilodalton glycoprotein with triglyceride lipase activity. Biol Reprod, 1997; 57(5): 1023-1031. Doi: 10.1095/biolreprod57.5.1023
30.    Malo C, Gil L, Gonzalez N, Cano R, de Blas I, Espinosa, E. Comparing sugar type supplementation for cryopreservation of boar semen in egg yolk based extender. Cryobiology 2010; 61(1): 17–21. Doi: 10.1016/j.cryobiol.2010.03.008.
31.    Molinia FC, Evans G, Quintana Casares PI, Maxwell W.M.C. Effect of monosaccharides and disaccharides in Tris-based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Anim Reprod Sci 1994; 36(1–2): 113–122. Doi:10.1016/0378-4320(94)90058-2
32.    Suksai M, Dhanaworavibul K. Effects of Trehalose and Sucrose on Human Sperm Motility, Vitality and Morphology after Cryopreservation. J Health Sci Med Res, 2019; 37(2): 101-107. Doi:10.31584/jhsmr.201945
33.    Aisen EG, Quintana M, Medina V, Morello H, Venturino A. Ultramicroscopic and biochemical changes in ram spermatozoa cryopreserved with trehalose based hypertonic extenders. Cryobiology 2005; 50(3): 239-249. Doi: 10.1016/j.cryobiol.2005.02.002 
34.    Chhillar S, Singh VK, Kumar R, Atreja SK. Effects of Taurine or Trehalose supplementation on functional competence of cryopreserved Karan Fries semen. Anim Reprod Sci, 2012; 135(1): 1-7. Doi: 10.1016/j.anireprosci.2012.08.029 
35.    Badr MR, Mary G, Hassan M.H. Effect of trehalose on cryopreservation, oxidative stress and DNA integrity of buffalo spermatozoa. J Reprod Infertil 2010;  1: 50-57.
36.    Stanishevskaya O, Silyukova Y, Pleshanov N, Kurochkin A. Role of Mono- and Disaccharide Combination in Cryoprotective Medium for Rooster Semen to Ensure Cryoresistance of Spermatozoa. Molecules. 2021; 29;26(19): 5920. Doi: 10.3390/molecules26195920. 
37.    Malo C, Gil L, Gonzalez N, Cano R, de Blas I, Espinosa, E. Comparing sugar type supplementation for cryopreservation of boar semen in egg yolk based extender. Cryobiology 2010; 61(1): 17–21. Doi: 10.1016/j.cryobiol.2010.03.008.
Volume 15, Issue 3 - Serial Number 32
This issue XML file is being prepared.
September 2023
Pages 7-15
  • Receive Date: 22 May 2023
  • Revise Date: 11 July 2023
  • Accept Date: 30 July 2023
  • First Publish Date: 01 September 2023