A comparison of equine spermatozoa viability using the TrueBreed+, TrueBreed and standard collection devices

Date

2022-08

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Abstract

ABSTRACT Previous research has demonstrated that the Device for Improved Semen Collection (DISC- marketed under TrueBreed™) helped maintain semen parameters for extended periods in a human and equine model compared to semen collected in a traditional collection vessel. By design, the DISC (1) slows the cooling rate, (2) limits exposed surface areas, (3) provides nutrients, and (4) lessens or eliminates osmotic shock during processing, theoretically protecting the cells from collection induced damage. The system was recently redesigned to include fixed scavengers to harvest reactive oxygen species (ROS). The objective of this study was to determine if this physiological redesign of the DISC protects sperm cell function in the equine model, using a series of biochemical assays to assess cellular function. Three semen samples were obtained from each of 10 stallions as part of a routine collection program. Each stallion was randomly collected one time in a standard container, the original equine TrueBreed, and the TrueBreed+ modified for antioxidant properties, using standard equine extension techniques. Once prepared, the samples were incubated at room temperature in the collection device, an evaluated holding temperature (20-23°C) to induce ROS. Sample aliquots were analyzed, and slides were prepared at 6, 9, 12, 24, 48, 72, and 96 hrs. Samples were assessed for standard motility parameters using a CASA instrument, and slides were prepared to assess morphology, acrosomes, mitochondrial function, and DNA fragmentation. The slides were prepared using standard commercial assays and a Biotek Cytation 5 with appropriate fluorescent filtration. Data were analyzed by ANOVA with repeated measures. While the study was designed to increase ROS generation, results indicate that stallion sperm cells collected in either the TrueBreed or TrueBreed+ maintained significantly better quality across all variables at all time points past six hours compared to the control (P < 0.05). This data suggests that more biochemically intact, physiologically active cells are found in samples collected and held in either the TrueBreed or TrueBreed+ devices. The results of this study continue to support previous studies supporting the role of the TrueBreed and TrueBreed+ systems in maintaining healthy sperm biochemistry and physiology. The present data demonstrate that the addition of ROS scavengers to the plastic matrix effectively protects sperm from DNA damage without adding anti-oxidant compounds to the media. Furthermore, the study suggests the TrueBreed+ system effectively protects stallion sperm cells from damage to the mitochondria and acrosome membranes at the time of collection. Future experimentation is necessary to determine the optimal scavenger to prevent cell damage. Lessening sperm damage in the sperm collection process provides the potential for improved sperm quality and potentially higher equine pregnancy rates.


Embargo status: Restricted until 09/2023. To request the author grant access, click on the PDF link to the left.

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Keywords

Spermatozoa, Equine

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