Effect of Fructose on Thawed Bull Semen’s Viability Obtained by Post-Mortem Collection
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2019, Vol 19, Issue 3
Abstract
During the cryopreservation process, bovine sperm cells become vulnerable, losing a large percentage of their motility and viability due to cryo-damages. Previous studies focused on the effect of fructose on other bovine species’ sperm viability indicated that the fructose application in the semen media enhanced the viability and motility of sperm cells. This survey aims to investigate the optimal concentration of fructose in the semen extender which is more effective for maintaining sperm viability after thawing. In the present paper, testicles of nine Hostein-Friesian bulls were recovered from a slaughterhouse, transported to the laboratory where sperm was collected from the epididymis’s tail. Immediately after collection, sperm was evaluated for pH, viability, motility and concentration. Then samples were submitted to cryopreservation process using different concentrations of fructose (5, 10 and 20mM – final concentration). After thawing, sperm was examined as afore described. Before freezing, no statistical differences were observed for the results obtained among bulls. On average, the pH and the motility were respectively 6.0±0.63 and 4.2±0.1. For the viability, in general, values ranged from 48.1% to 77.8% (X=65.0%±2.9). After thawing, besides no statistical differences were observed among bulls, they have been observed among treatments. When compared to the control, results of viability increased on the concentration of 10mM of fructose from 41.2%±3.7 to 47.0±3.8, respectively for the sperm belonging to the control and the 10mM group, while no significant differences were observed for the motility values between these treatments (2.8±0.2). At the concentration of 20mM, a decrease of sperm viability was observed (38.2%±3.8) when compared with the control (P>0.05). A slight decrease was also observed in the motility, in which values decreased to 2.7±0.1. Results of the present study indicated that adding low concentrations of fructose in the extender (10mM) could improve the viability of the sperm cells after thawing. So, fructose is considered to be an efficient energy source for sperm cells who go through the cryopreservation process.Semen cryopreservation is a well-established procedure used in veterinary and human assisted reproduction technology applications. Over the last 50 years, it was used for genetic improvement of beef and dairy cattle. Moreover, it helps the regulation of venereal diseases and facilitate the management of cattle fertility. Normally semen is recovered from alive animals, but in some situations e.g. when the male succumb the only possibility is recovering semen from the epididymis. This portion of the testicle is usually divided into four anatomical regions: the initial segment, head (caput), body (corpus), and tail (cauda). The epididymis is organized into lobules separated by connective tissue septa that serve not only as internal support for the organ but also as a functional separation which allows for selective expression of genes and proteins within each individual lobule [1,2]. The most important factors for successful semen cryopreservation according to various surveys are, the composition of the extender, the suitable cryoprotectants and the optimal freezing and thawing rates [3]. Spermatozoa are characterized by plasma membrane fluidity and low water content which make them more resistant to cryo-damage compared to other cell types such as oocytes [4]. However, cryo-damage can happen as a consequence of thermal stress due to the change in temperature during cooling, freezing and thawing as well as the osmotic stress caused by addition of high concentrations of cryoprotective agents and by crystallization [5]. This results in protein denaturation, shrinkage and irreversible membrane collapse [6]. Therefore, phospholipids and cryoprotective agents, optimal dilution, equilibration and cooling procedures are required to avoid cold shock so as to reduce crystallization and minimize sperm damage [7]. It is well known that a structurally and biochemically active plasma membrane is required to accomplish the processes of capacitation, acrosome reaction and the oocyte penetration. Semen extender is needed to maintain the sperm motility. Spermatozoa require the proper nutrition for their viability. Fructose is a type of sugar which can be easily metabolized into energy. Its addition, into the extender can be a main source of energy for spermatozoa in the seminal plasma for metabolic processes [8]. The use of sugar such as fructose, sucrose, glucose, trehalose, EDTA or rafinose in semen extender, could increase sperm motility and viability (Aisen; Suwarso, 1999 in Arifiantini). Sugar in semen extender could keep osmotic pressure and energy source on specific levels as a cryoprotectant.
Authors and Affiliations
Athanasia Zoi Pappa, Helena Moreira da Silva, Loide Valadão, Fernando Moreira da Silva
Keywords
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- EP ID EP621761
- DOI 10.26717/BJSTR.2019.19.003300
- Views 178
- Downloads 0
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