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FROZEN-THAWED QUALITY OF SWAMP BUFFALO SPERMATOZOA CRYOPRESERVED WITH SUGAR PALM JUICE EXTENDER

FROZEN-THAWED QUALITY OF SWAMP BUFFALO SPERMATOZOA CRYOPRESERVED WITH SUGAR PALM JUICE EXTENDER

RIZAL, MUHAMMAD; RIYADHI, MUHAMMAD; YULNAWATI, YUSNIZAR
URI: https://repo-dosen.ulm.ac.id//handle/123456789/19037
Date: 2020-09-30

Abstract:

The objective of this research was to examine the effectivity of sugar palm juice as an alternative extender to cryopreserve buffalo. Moreover, we expected to obtain the best composition of sugar palm juice that can be used widely as an alternative extender for buffalo semen cryopreservation. Semen was collected from swamp buffalo bull using artificial vagina. Fresh, diluted, equilibrated, and frozen-thawed was evaluated for general parameters, i.e spermatozoa progressive motility (M), viability (V) and membrane integrity (MI). Fresh semen that had good quality was diluted up to 1.108 sperm/ml in four different compositions of extenders, i.e lactose extender containing 20% egg yolk as control (C), 74% sugar palm juice + 20% egg yolk + 6% glycerol (SPG6), 73% sugar palm juice + 20% egg yolk + 7% glycerol (SPG7), and 72% sugar palm juice + 20% egg yolk + 8% glycerol (SPG8), respectively. Diluted-semen was equilibrated at 5oC for 4 h. It followed by frozen and stored in liquid nitrogen (-196oC). The results showed that the percentages of M, V, and MI of frozen-thawed spermatozoa in control (C) were 45.83, 59.37, and 57.83%), respectively. These results were significantly (P<0.05) higher than those parameters in SPG7 (30.83, 38.83, and 48.83%), SPG6 (18.33, 38.83, and 40.33%), and SPG8 (18.33, 39.17, and 39.33%). In conclusion, sugar palm juice might not be as ideal as control group in maintaining frozen-thawed swamp buffalo semen quality. But this inform us that in a very remote area where chemical or commercial extenders are not easy to find, sugar palm juice can be an alternative due to the fact that it did not affect the pregnancy rate of AI program. Moreover, the cryoprotectant (glycerol) concentration play important role in order to maintain the spermatozoa post thawed quality.

Description:

Artificial insemination (AI) is most common reproductive technologies that is very useful to increase farm animals population. It is also useful for genetic improvement from the male side (Harshan et al., 2006). AI technology is an integrated process of semen processing (preservation or cryopreservation) and the artificial insemination itself. In most effective end eficient way, AI has been done using frozen-thawed semen. In the cryopreservation process, ice crystal formation can be harmful for sperm viability. Ice crystal formation occurs during the frozen stage, where the spermatozoa cells exposes to very low temperature (-196oC) of liquid nitrogen. Intracellular ice crystallization during cryopreservation is the main cause of damage to the cells. Due to ice crystal formation, normally about 50% of spermatozoa are damaged during cryopreservation (Watson, 2000). Therefore, extender composition and suitable cryoprotectants are important factors for successful semen cryopreservation (Hammerstedt et al., 1990; Curry et al., 1994). The cryoprotectant agent is essential for the viability of spermatozoa after thawing, as these agents minimize intracellular ice formation and restrict the solution effect (Medeiros et al., 2002). Addition of intracellular cryoprotectant compounds, cause increased extender osmotic pressure, and potentially damage spermatozoa cells. Water will leaves of the cell so that spermatozoa shrink, then swell when cryoprotectant and water reenter the cell to maintain chemical equilibrium. At the time of cryoprotectant removal, the cell initially will swell due to the influx of water and then slowly return to the isoosmotic state when cryoprotectant and water leave the cell. Repeated changes in osmotic pressure can result in significant loss of spermatozoa functional integrity, such as motility, or even cell death without loss of plasma membrane integrity (Gao et al., 1997). Since the discovery of glycerol (Polge et al., 1949), it has been used extensively for the cryopreservation of many types of cells, including mammalian spermatozoa (McGonagle et al., 2002). However, glycerol has osmotic and toxic effects on the plasma membrane and metabolism of cryopreserved cells (Hammerstedt et al., 1990). It is responsible for the disorganization of spermatozoa plasma membrane (Hammerstedt and Graham, 1992; Buhr et al., 2001) and reducing motility and fertilizing ability (Jeyendran et al., 1985). Higher concentrations of glycerol lead to cell death (Wündrich et al., 2006).

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