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Abstract
BackgroundVariability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics).ResultsPost-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05).ConclusionsThese results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics.
Highlights
Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method
The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols, developed at our laboratory [21] in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times
Significant differences were seen between stallion donors in terms of mean Sperm DNA fragmentation index (sDFI) (p
Summary
Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics). Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method [1,2]. Spermatozoa with damaged chromatin appear normal in terms of their membrane integrity, morphology and motility, but their use would lead to post-fertilization embryo failure [5]. The analysis of DNA fragmentation dynamics is important since DNA can become fragmented during this time, even within the female reproductive tract
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