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. 2017 Nov;24(7):1489-1496.
doi: 10.1016/j.sjbs.2016.01.030. Epub 2016 Jan 22.

In vivo fertilizing ability of stallion spermatozoa processed by single layer centrifugation with Androcoll-E™

Sandra Gamboa  1   2 , Andreia Quaresma  1 , Fátima Castro  1 , Pedro Bravo  1 , Maria Rosa Rebordão  1 , Maria do Mar Oom  3   4 , António Rocha  5
Affiliations

In vivo fertilizing ability of stallion spermatozoa processed by single layer centrifugation with Androcoll-E™

Sandra Gamboa et al. Saudi J Biol Sci. 2017 Nov.

Abstract

A colloid with a species specific silane-coated, silica-based formulation, optimized for stallion (Androcoll-E™), enables a better sub-population of spermatozoa to be selected from stallion ejaculates. However, such a practice has not been critically evaluated in stallions with fertility problems. In this study we evaluate whether single-layer centrifugation (SLC) through Androcoll-E™ could be used to enhance fertility rates in a subfertile stallion. Ejaculates were obtained from two different stallions, one Lusitano (fertile) and one Sorraia (subfertile), with distinct sperm characteristics and fertility. Motility, morphology, plasma membrane structural (eosin-nigrosin) and functional integrity (HOS test), mitochondrial functionality (Δψm; JC-1) and longevity (motility after 72 h cooling) after centrifugation in Androcoll-E™, as well as pregnancy rates obtained after artificial insemination (AI), with and without (control group) SLC-treated sperm were assessed. The effect of SLC on sperm characteristics, and fertility results were evaluated by ANOVA and Fisher procedures, respectively. Our results showed that SLC-selected sperm did not differ from the raw semen in terms of viability, morphology, response to hypo-osmotic conditions (HOS test) and mitochondrial membrane potential (↑ΔΨmit; JC-1). Sperm motility in cooled samples was not improved by SLC treatment. Our data show that SLC through Androcoll-E™ has no effect on pregnancy rates in the stallions used in this trial.

Keywords: Fertility; HOS test; Mitochondria; Single-layer centrifugation; Sperm; Stallion.

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Figures

Figure 1
Figure 1
Schematic representation of the processing of the ejaculates using the single-layer centrifugation (SLC) with Androcoll-E™, respectively for Lusitano (PSL; left panel) and Sorraia (right panel) sperm. The position of the different layers are shown (black arrows).
Figure 2
Figure 2
Sperm characteristics in raw semen and in the different layers after SLC-treatment. Proportion (mean ± SEM) of sperm (A) vitality, (B) morphology, (C) HOS+, abnormalities in (D) head, (E) midpiece (MP) and (F) principal piece (PP). Different superscript in mean values represents significant differences (P < .05) between layers (a, b) and between stallions (*).
Figure 3
Figure 3
Proportion (mean ± SEM) of sperm mitochondrial membrane potential (ΔΨmit) in raw semen and in the different layers after single-layer centrifugation (SLC) on the sperm from Lusitano (A) and Sorraia (B) stallions. ↑ΔΨmit, high mitochondrial membrane potential; ↓ΔΨmit, low mitochondrial membrane potential; w/ΔΨmit, without mitochondrial membrane potential. Values bearing + possible differ statistically (P < 0.1).
Figure 4
Figure 4
Proportion (mean) of motile spermatozoa after collection (PMSAC) and after dilution in IRA96, with (SLC-selected) and without (non SLC-selected) centrifugation on Androcoll-E™ , cooled (4 °C) and stored for 24 h (PMS24 h), 48 h (PMS48 h) and 72 h (PMS72 h). Values bearing * differ significantly between stallions (P < 0.05).
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