The effect of the zirconia structure in Ni/ZrO2 catalysts on the glycerol steam reforming (GSR) reaction was studied. A tetragonal zirconia support was synthesized via a hydrolysis technique and loaded with 5 wt% Ni via a wet-impregnation method. Similarly, a commercial monoclinic zirconia support was also impregnated with 5 wt% Ni. Following calcination at 600 °C, physico-chemical properties of the prepared catalysts were investigated by X-Ray Diffraction (XRD), H2-Temperature Programmed Reduction (H2-TPR) and CO2-Temperature Programmed Desorption (CO2-TPD) techniques. The catalysts were then tested in the GSR reaction in the 400–700 °C range with a steam to glycerol molar ratio of 9:1 and a flow rate of 0.025 mL/min. The monoclinic catalyst exhibited a better performance giving higher hydrogen yields and glycerol conversions. This was attributed to an improved reducibility of Ni in this catalyst. Stability tests at 600 °C revealed the deactivation of the tetragonal catalyst during 6 h as a result of the formation of encapsulating coke which blocked active Ni metal sites. The monoclinic catalyst, exhibiting the formation of only filamentous coke, remained relatively stable for 24 h.