Quality of Garut Ram’s Sperm in Tris Extender Supplemented with Ethylene Diamine Tetraacetic Acid Preserved at Low Temperature (3–5°C)

Abstract

Ethylenediaminetetraacetic acid (EDTA) is a chemical substance that can bind the heavy metal and other toxic compounds so it is suitable to use for protecting sperm during preservation at 3-5°C. The research aimed to evaluate the role of EDTA to maintain the quality of Garut ram sperm in low-temperature preservation. Semen was collected by an artificial vagina. Fresh semen was evaluated and divided into 3 tubes and diluted with tris extender contain 20% egg yolk (TEY) without EDTA (T1), TEY+0.01% EDTA (T2) and TEY+0.02% EDTA (T 3). Semen was evaluated both macroscopically and microscopically before preserved in a refrigerator at 3-5°C. During storage, semen was also evaluated on daily basis (four consecutive days) that included variables relate to sperm motility (SM), sperm viability (SV), sperm membrane integrity (SMI) and sperm acrosome intact (SAI). The research design used was Completely Randomized Design with three treatments and five replications. Differences between treatments were tested with the least significant difference test. The results showed that the average of each variable gained of fresh semen was 0.88mL for volume, cream for color, thick for consistency and 7 for pH. While the mass movement of sperm was +++ and sperm concentration was 3,962 million cells/mL. The average percentages of SM, SV, abnormality, SMI, and SAI were 75.00%, 86.60%, 4.80%, 84.40%, and 84.80%, respectively. On the day-4 of storage, percentages of SM, SV, SMI, and SAI in control (T1) decreased to 48.75%, 66.25%, 64.75%, and 63.5%, respectively, but still met the requirement of sperm quality for artificial insemination purposes. Moreover, there was no significant difference between T2 (45.00%, 65.00%, 63.25% and 65.25%) and T3 (40.00%, 65.67%, 60.75% and 62.50%) of those variables measured. Finally, it was concluded that the addition of EDTA in Tris extender could maintain the quality of Garut ram sperm.