Chilling process has negative impact on sperm function and quality attributes resulting in some modifications at the biochemical and structural levels. Selenium nanoparticles (SeNPs) supplementation could stop or even reverse this impact and improve the quality of cooled rabbit. The present study was designed to evaluate the effects of adding five different concentrations of SeNPs (0.5, 1, 1.5, 2 and 2.5 µg/ml) to protect rabbit spermatozoa from chilling adverse effects. X-ray diffraction (XRD), high resolution transmission electron microscopy (HR-TEM), and scanning electron microscopy (SEM) were used to characterize SeNPs synthesized by co-precipitation method. Semen was collected from ten sexually mature and fertile New Zealand White male rabbits using artificial vagina and pooled semen samples were diluted with tris-egg yolk-fructose extender only (control) or extender supplemented with 0.5, 1, 1.5, 2 and 2.5 µg/ml of SeNPs. Sperm motility, membrane integrity and acrosome defects were measured. To determine antioxidative role of SeNPs, malondialdehyde (MDA) level as a biomarker of lipid peroxidation was measured in seminal plasma. Sperm motility and membrane integrity increased with SeNPs at concentrations 0.5 μg/ml. Likewise, no adverse effect was reported on percentages of a live sperm as well as sperm morphology with all concentrations. MDA levels decreased with concentration 0.5 μg/ml SeNPs. Notably, adding 0.5, 1 and 2 μg/ml SeNPs to extender produced best significant positive effects on semen parameters compared to SeNPs free group. In conclusion, the results confirm that low concentrations of SeNPs fabricated by co-precipitation method could effectively preserve the quality of chilled rabbit semen.