# The effects of supplementation of vitamin D to the egg‐yolk extender on cryopreservation of ram semen

## TL;DR

Adding vitamin D to a freezing solution improved the quality of frozen ram semen, with 50 ng/mL being the most effective concentration.

## Contribution

This study demonstrates that 50 ng/mL vitamin D in an extender improves ram semen cryopreservation outcomes.

## Key findings

- Sperm motility and functional integrity were highest at 50 ng/mL vitamin D.
- Vitamin D at 50 ng/mL improved mitochondrial membrane potential and sperm straightness.
- The 50 ng/mL dose provided better protection than other concentrations during freezing.

## Abstract

This study aimed to examine the effects of supplementation of vitamin D to the egg‐yolk extender on characteristics of frozen–thawed ram semen.

Semen samples obtained from adult rams were pooled and divided into five equal volumes. It was reconstituted with extenders containing different concentrations of vitamin D: 0 (control), 12.5 (VITD 12.5), 25 (VITD 25), 50 (VITD 50), and 100 ng/mL (VITD 100), and then they were frozen. Sperm motility parameters, plasma membrane functional integrity, acrosomal integrity, DNA fragmentation, and mitochondrial membrane potential of the groups were evaluated after sperm thawing.

Total motility and progressive motility were higher in VITD 50 than in all other groups (p < 0.05). Higher sperm straightness, linearity, and wooble were higher in VITD 50 than in the control group (p < 0.05). A similar pattern of VITD 50 was observed for plasma membrane integrity and mitochondrial membrane potential (p > 0.05).

In the study, it was observed that adding vitamin D to the extender had a beneficial effect on ram spermatological parameters. In addition, it was concluded that the use of the 50 ng/mL vitamin D in the extender provided more effective protection than the other doses.

This study aimed to examine the effects of supplementation of vitamin D to the egg‐yolk extender on characteristics of frozen–thawed ram semen. Sperm motility, plasma membrane functional integrity, acrosomal integrity, DNA fragmentation, and mitochondrial membrane potential of the groups were determined. In the study, it was observed that adding vitamin D to the extender had a beneficial effect on ram spermatological parameters. In addition, it was concluded that the use of the 50 ng/mL vitamin D in the extender provided more effective protection than the other doses.