Effects of Double Layer Centrifugation on the Improvement of Sperm Quality in Dogs: A Comparative Note among Different Breeds

Abstract

The evaluation of sperm quality in the laboratory is essential to improve efficiency in assisted reproduction. As in other species, for the dog there are reports that prolonged contact of sperm with some components of seminal plasma is associated with decreased motility and sperm viability. Thus, the centrifugation is a technique widely used to concentrate the spermatozoa and eliminate the supernatant. The purpose of this study was to evaluate the effect of double layer centrifugation on the percentages of total sperm motility and progressive sperm motility of the dog’s semen submitted to the dilution, single layer centrifugation, cooling and storage at 5 °C for 24 and 48 hours. For this purpose, ejaculates of 30 healthy male dogs were evaluated, by taking into account the comparison among the conventional sperm parameters (ejaculate volume, sperm concentration, total sperm motility and sperm progressive motility). The semen samples were examined in standard baseline condition of fresh semen (FS), after dilution (AD), after dilution and single layer centrifugation (SLC), after double layer centrifugation (DLC). According to the different time points, the semen samples were evaluated in baseline conditions, immediately after their collection at (T0), at 24 h (T24) and at 48 h (T48), to evaluate the effect of different treatments on the semen’s quality. Results showed a significant effect of double layer centrifugation on the improvement of total sperm motility and progressive sperm motility percentages of dogs. The use of cooling fresh semen soon after the double layer centrifugation will improve the semen quality up to 48h, with a special emphasis for the percentages of total sperm motility and sperm progressive motility, adding an alternative technical approach to reproductive performance in male breeding dogs.