Objective-To investigate the effects of ketamine hydrochloride on the analgesic effects of tramadol hydrochloride in horses with signs of pain associated with naturally occurring chronic laminitis. Animals-15 client-owned adult horses with chronic laminitis. Procedures-Each horse received tramadol alone or tramadol and ketamine in a randomized, crossover study (≥ 2 months between treatments). Tramadol (5 mg/kg) was administered orally every 12 hours for 1 week. When appropriate, ketamine (0.6 mg/kg/h) was administered IV for 6 hours on each of the first 3 days of tramadol administration. Noninvasive systemic blood pressure values, heart and respiratory rates, intestinal sounds, forelimb load and off-loading frequency (determined via force plate system), and plasma tumor necrosis factor-α and thromboxane B2 concentrations were assessed before (baseline) during (7 days) and after (3 days) each treatment. Results-Compared with baseline data, arterial blood pressure decreased significantly both during and after tramadol-ketamine treatment but not with tramadol alone. Forelimb off-loading frequency significantly decreased during the first 3 days of treatment with tramadol only, returning to baseline frequency thereafter. The addition of ketamine to tramadol treatment reduced off-loading frequency both during and after treatment. Forelimb load did not change with tramadol alone but increased with tramadol-ketamine treatment. Plasma concentrations of tumor necrosis factor-α and thromboxane B2 were significantly reduced with tramadol-ketamine treatment but not with tramadol alone. Conclusions and Clinical Relevance-In horses with chronic laminitis, tramadol administration induced limited analgesia, but this effect was significantly enhanced by administration of subanesthetic doses of ketamine.