Objective Analysis of any effect of retrobulbar block during ocular surgery on heart rate variability and oculocardiac reflex. Study design Prospective study. Animal studied Horses (n=16) undergoing eye enucleation due to chronic ophthalmologic diseases. Procedure Eye enucleation was performed under general anesthesia. The horses were randomly assigned to the first (inhalation anesthesia only, n=10) or second group (inhalation and local retrobulbar anesthesia, n=6). The retrobulbar block was performed using 12mL of mepivacaine hydrochloride 2%. ECG data were taken by a Telemetric ECG before, during, and after surgery. Heart rate variability was analyzed in the time domain as mean heart rate, mean beat-to-beat interval duration, and standard deviation of continuous beat-to-beat intervals. The frequency domain analysis included the low- and high-frequency components of heart rate variability and the sympathovagal balance (low/high frequency). The low frequency represents mainly sympathetic influences on the heart, whereas high frequency is mediated by the parasympathetic tone. Results All horses without a retrobulbar block showed a significant decrease in the heart rate during traction on the globe and pressure on the orbital fat pad for homoestasis (P=0.04). Simultaneously, high-frequency power, as an indicator of vagal stimulation, increased significantly. High-frequency and low-frequency power in the retrobulbar block group increased in five horses, and heart rate decreased in only one horse. Both were not significant within the group, but there was a significant difference between both groups relating to the incidence of heart rate decrease occurring at globe traction. Conclusions and clinical relevance Heart rate variability is a sensitive, non-invasive parameter to obtain sympathovagal stimulations during general anesthesia. The retrobulbar block can prevent heart rate decrease associated with initiation of the oculocardiac reflex.