Abstract
The objective of this study was to scale-up the procedure for Single Layer Centrifugation (SLC) through AndrocollTM-P, as a preliminary step towords processing the whole ejaculate. The first experiment compared Single Layer Centrifugation using 4.5 mL and 15 mL extended ejaculate (SLC-4.5 and SLC-15, resp.), assessing sperm quality by objective motility analysis, morphology, viability, and the production of reactive oxygen species (ROS). In the second experiment, SLC-4.5 was compared to Single Layer Centrifugation with 25 mL extended ejaculate (SLC-25) using motility analysis and morphology. In both experiments, normal morphology and linear motility were significantly higher in the SLC-selected samples than in the uncentrifuged controls (P < .001), whereas total motility and membrane integrity were unchanged. Although ROS production was higher in the SLC-selected samples than in the controls (P < .01), this might have been due to the presence of antioxidants in seminal plasma in the latter. In conclusion, there was no difference in sperm quality between SLC-4.5 and SLC-15 samples, or between SLC-4.5 and SLC-25 samples, indicating that the SLC method can be scaled-up successfully.
Highlights
A technique for improving sperm quality is considered desirable in many domestic species, for example, cattle and horses, where sperm quality is related to pregnancy rates after artificial insemination (AI)
The objective of this paper was to scale up the original Single Layer Centrifugation (SLC) method to allow larger volumes of ejaculate to be processed
[11] in the present study, less bovine serum albumin (BSA) (1.25 mg/mL) was added to the Beltsville Thawing Solution (BTS) in Experiment 1 compared to previous studies, which resulted in spermatozoa adhering to the microscope slide, not being included in the Computer-Assisted Sperm Motility Analysis (CASA) motility analysis, and might have contributed to decreased sperm survival
Summary
A technique for improving sperm quality is considered desirable in many domestic species, for example, cattle and horses, where sperm quality is related to pregnancy rates after artificial insemination (AI). A method that selects the most robust spermatozoa from an ejaculate could have many advantages for the boar AI industry, either by reducing the sperm number used for AI [5], potentially enhancing sperm survival during cryopreservation, or by removing pathogens in the ejaculate such as bacteria [6] and viruses [7]. Such a selection method would have a cost benefit for the pig industry, depending on the value of the boar, the actual price of semen doses, and the perceived need to increase biosecurity and reduce antibiotic usage
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