Development of a frozen bovine semen diluent: Comparison of particle size of commercial and home-made diluents

Abstract

A variety of diluents, both homemade and commercial, are used in Indonesian artificial insemination centers to freeze bovine semen. One of the homemade diluents used is Tris egg yolk. The egg yolk is used as a source of lecithin to protect the sperm membrane during freezing. The smaller the lecithin particles, the more effectively they can bind to the cell membrane. In addition, the particle size facilitates the analysis of sperm quality using Computer Assisted Sperm Analysis (CASA). This study aimed to perform particle analysis of lecithin in homemade and commercial diluents using a particle size analyzer (PSA). The diluents used in this study were Tris egg yolk, Andromed (Minitube, Germany), Bovifree (Minitube, Germany), OptiXcell (IMV, France), and BioXcell (IMV, France). The results showed that the Tris egg yolk diluent had the widest particle size range, from 584.5 ± 2.2 to 3255.7 ± 48.84 nm. The smallest commercial diluent was OptiXcell (64.6 ± 2 nm), followed by BioXcell (186.2 ± 2.2 nm), Andromed (245.7 ± 2.8 nm) and Bovifree (278.5 ± 2.5 nm). The sperm kinematics of frozen semen in Tris egg yolk diluent were challenging to analyze. In contrast, the kinematics of semen in commercial dilutions could be visualized and evaluated according to the established criteria. This studys results indicate that the commercial diluents particle size is smaller than that of the homemade diluent and does not affect the results of the CASA assessment. Consequently, developing homemade diluents with a particle size comparable to commercial diluents is essential.