Abstract
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.
Highlights
Most equine artificial inseminations (AI) are performed with cooled semen rather than with fresh semen immediately after collection [1]
Each ejaculate was split among the following treatments: (1) single-layer centrifugation (SLC) in INRA82 (A1); (2) SLC in INRA96 (A2); (3) sperm washing in INRA82 (B1); (4) sperm washing in INRA96 (B2); (5) extension in INRA82 (C1), (6) extension in INRA96 (C2)
Sperm motility was higher if processed by SLC than by sperm washing or extension, except for sperm washing in INRA96 at 0 and 96 hours
Summary
Most equine artificial inseminations (AI) are performed with cooled semen rather than with fresh semen immediately after collection [1]. Removing most of the seminal plasma (SP) from stallion ejaculates could have a beneficial effect on stabilizing sperm membranes during cooled storage [2] and might reduce chromatin damage [3], presumably by removing sources of reactive oxygen species. It may reduce the inflammatory response in mares prone to post-breeding endometritisdan exaggerated form of the normal inflammatory response in the uterus [4,5]. Animal welfare/ethical statement: Animals were housed and handled according to national and institutional regulations for animal care and use
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