Abstract
This study was designed to develop a method of liquid storage of boar sperm at 4°C by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N- acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at 4°C for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at 4°C. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to 20×10 5 sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of 2.5×10 5 sperm/ml was lower than in concentrations of 5, 10 and 20×10 5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at 2.5×10 5 /ml sperm concentration was significantly lower than at 5, 10 and 20×10 5 sperm/ml concentrations. In conclusion, GPL4 diluent can be stored at 4°C for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes. (Key Words : Fertilization, Liquid Semen, Oocyte, Porcine)
Highlights
The storage technologies of boar semen can be divided into two major types: the liquid storage and the cryopreservation
The concentrated sperm was resuspended with 10 ml of lactose-egg yolk and Nacetyl-D-glucosamine (LEN), BF5 or the modified BF5 with bovine serum albumin (BSA) and N-acetyl-Dglucosamine (GPL4) diluents using for frozen boar semen processing to provide 1.0×109 sperm/ml at room temperature
Our results clearly demonstrated that the modified BF5 diluent with N-acetyl-D-glucosamine and BSA (GPL4) is efficient for storage of boar sperm at 4°C for 5 days
Summary
The storage technologies of boar semen can be divided into two major types: the liquid storage and the cryopreservation. According to the range of temperature, the liquid storage can be divided into the normal temperature storage (15-25°C, the short term storage) and the low temperature storage (0-5°C, the long term storage). Because of their peculiar plasma membrane composition, boar sperm are extremely sensitive to cooling, freezing and thawing (White, 1993). The primary action of BSA in an extended sperm suspension is surely a stimulation of the sperm motility, which appears to be reversible. Various methods have been employed for the evaluation of motility, acrosome integrity, sperm viability and sperm ATP concentration. Sellés et al (2003) reported
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