Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4°C

An extender has been developed with low-density lipoproteins (LDLs) that eliminates the microbial risks associated with the use of whole egg yolk. The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 degrees C, as well as on spermatozoa motility, plasma membrane and acrosome integrity, at two different concentrations (80 x 10(6) and 240 x 10(6) sperm per ml) for 8 days and to evaluate glycerol toxicity in both extenders. A total of 12 ejaculates were collected from three bulls. Spermatozoa motility was examined using computer-assisted semen analysis. Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test. The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol (TEG), Tris-egg yolk without glycerol (TE), LDL with glycerol (LDL+) and LDL without glycerol (LDL-), at 80 x 10(6) and 240 x 10(6) sperm per ml. This study showed that the LDL 1 and LDL 2 extenders were more effective at preserving spermatozoa motility, plasma membrane integrity and acrosome integrity than TEG and TE (P < 0.05) during 8 days of incubation. After 3 days of incubation, a toxicity of glycerol was observed in TEG, whereas no significant difference was observed between LDL+ and LDL-. We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 degrees C instead of TEG and TE at 80 x 10(6) and 240 x 10(6) sperm per ml for elite bulls. This finding can be used to define a policy for the storage of high-quality bull semen. Asian Journal of Andrology (2011) 13, 281-286; doi:10.1038/aja.2010.84; published online 29 November 2010