Disbudding of calves is a common, painful intervention. Due to cytotoxic and anesthetic properties, the injection of clove oil or its component isoeugenol may be less detrimental to animal welfare. We investigated mechanical nociceptive threshold (MNT), possible tissue alterations and horn growth for up to 12 weeks after injection of 1.5 mL clove oil (CLOV), isoeugenol (ISO) or saline (CON) or after hot-iron disbudding (BURN; with local anesthesia and sedation, n = 10/treatment). MNT was measured using von Frey filaments and a pressure algometer at four locations around the horn bud. There was a treatment*time point interaction (linear mixed model, p< 0.05). MNT decreased most strongly and for the longest time for BURN in most calves at least for 3 weeks. For ISO, the decrease was less distinct and most calves' values returned to baseline after 1-2 weeks. MNT in CLOV was intermediate, with decreased values up to 3 weeks in some animals. 12 weeks after the treatment, horn growth was prevented in about 50% of the horns in CLOV and ISO. Tissue alterations such as swellings of the eyelids often occurred in CLOV, but less so in ISO. Our results suggest that injection of isoeugenol causes less pain and thus seems to be beneficial compared to hot-iron disbudding, while clove oil was not advantageous. Regarding the effectiveness of isoeugenol to prevent horn growth, more studies are needed.