Stored Stallion Sperm Quality Depends on Sperm Preparation Method in INRA82 or INRA96

Removal of seminal plasma facilitates stallion sperm survival during storage, but washing may damage sperm chromatin. Therefore, sperm quality was compared in samples following single-layer centrifugation (SLC) or sperm washing and controls (extension only) in two extenders, INRA82 and INRA96. Ejaculates from six stallions were split among six treatments: SLC, sperm washing, and controls, in INRA82 and INRA96. Sperm motility and acrosome status were evaluated at 0, 24, 48, 72 and 96 hours; morphology at 0, 24, 48, 72 hours and chromatin integrity at 0 and 96 hours, with storage at 6°C. Sperm samples in INRA96 had better motility, acrosome status, and normal morphology than samples in INRA82. The SLC samples had higher motility and fewer reacted acrosomes than controls, and lower fragmented chromatin than washed samples. Fewer spermatozoa with tail defects were observed after SLC than after sperm washing; spermatozoa washed in INRA82 had fewer tail defects than those washed in INRA96. In conclusion, sperm quality (except for morphology) was better in INRA96 than in INRA82 and was better in SLC samples than in washed samples or controls. The SLC method is a useful adjunct to stallion sperm preparation, especially for storage before artificial insemination.