The Motility Ratio method as a novel approach to qualify semen assessment

Many scientific studies often assumed that the most reliable methods for assessing sperm motility are those that give the highest values, and this leads to misinterpretation of the results. This study aims to propose an objective method to validate sperm motility reliability. Bovine and porcine semen samples were split into two equal fractions. Fraction A was kept alive with a motile population considered at maximum proportion, while fraction B was killed with 0% motile population. A range of motile/non motile sperm was performed by mixing both fractions. The Motility Ratio method, comparing measured and theoretical motility, was validated using LEJA slide and IVOS II and applied to other slides. All slides showed strong Concordance Correlation Coefficient between measured and theoretical motility. However, with IVOS II, LEJA slide showed the lowest bias (< 1) while MAKLER or coverslip showed higher bias (> 2 and > 7 respectively) between measured and theoretical motility. This study shows that the best sperm motility is not always the true motility and highlights the importance of implementing a gold standard methodology for motility reliability such as The Motility Ratio method.