In standing ophthalmic surgery in horses, a retrobulbar nerve block (RNB) is often placed blindly for anesthesia and akinesia. The ultrasound (US)-guided RNB may have fewer complications, but the two techniques have only been compared once in equine cadavers. This study compares the techniques for success and complication rates and analyzes the effect of training on US guidance. Twenty-two equine cadavers were divided into three groups: blind RNBs were performed bilaterally in eight cadavers, US-guided RNBs were performed bilaterally in seven cadavers, and after US-guided training, blind RNBs were performed bilaterally in seven cadavers. All RNBs were performed by the same two inexperienced operators, and a combination of contrast medium (CM; 1.25 mL) and methylene blue dye (1.25 mL) were injected (2.5 mL total volume). Needle positioning in the periorbita and the distance of the CM to the optic foramen were assessed using computerized tomography (CT). Dye spreading was evaluated by dissection. In group 1, 37.5% of the injections were in the optimal central position in the periorbita; in group 2, 75% and in group 3, 71.4%. There was no significant difference between the groups regarding needle position (groups 1 and 2 p = 0.056; groups 1 and 3 p = 0.069, groups 2 and 3 p = 0.8). The mean CM distribution distance was not significantly different between all groups. Group 1 had 18.75% intraocular injections versus 0% in group 2 and 7.1% in group 3 (not significant). US guidance showed no significant increases in accuracy nor decreases in complications. However, the effects on accuracy showed a trend towards significant improvement, and larger scale follow-up studies might show significant training effects on US guidance.