In many cell types Mg2+ can antagonise Ca2+ -stimulated signalling pathways, but information regarding the effects of these ions on IVF and subsequent embryonic development is limited. Our objectives were to evaluate the effects of Mg2+ in the IVF medium on embryonic development in mice and then determine if similar effects occurred in domestic cats and humans. Oocytes from hybrid and outbred mice, domestic cats and humans were fertilised (IVF, mice and cats; intracytoplasmic sperm injection (ICSI), humans) in the presence of 0.2 or 1.2 (mouse and human) or 1.0 (cat) mM Mg2+ and the resulting embryos were cultured to the blastocyst stage. Decreased concentrations of Mg2+ during IVF increased (P<0.05) cleavage of oocytes from outbred mice (77.9 vs. 51.0%), development of embryos from hybrid mice (74.5 vs. 51.0% hatching blastocyst per cleaved embryo) and both cleavage (68.4 vs. 46.8%) and blastocyst development (53.0 vs. 26.2% per cleaved embryo) in cats. Development to the blastocyst stage (52.1 vs. 40.2%) was also improved (P<0.05) when ICSI was performed on human oocytes in the presence of 0.2 mM Mg2+, compared with a commercial culture medium. Sensitivity to increased (1.0 to 1.2 mM) concentrations of Mg2+ in the medium during the oocyte-to-embryo transition appears to be conserved in three different species.