The aim of this experiment was to evaluate the effect of undenatured type II collagen supplementation on inflammation and pain using gait analysis and industry-accepted pain and mobility questionnaires during an exercise regimen in healthy dogs. Forty healthy Labrador Retrievers (20 male/20 female; Range: 5-12yrs) were sorted into two groups: undenatured type II collagen group receiving 40mg UC-II product (10mg total collagen and ≥3% undenatured type II collagen) and placebo group receiving 40mg maltodextrin daily by capsule. After 2wks loading, all dogs began an 11wk endurance exercise regimen consisting of two weekly runs, starting at 5km and increasingly incrementally to 8km, with one final 16km run. Gait analysis was performed at baseline; before, 24h and 48h after the first 5km run; and before, 24h and 48h after the final 16km run. Gait analysis was calculated to obtain a Four Rivers Kennel (FRK) Inflammation Index score. Dogs were scored according to the Liverpool Osteoarthritis in Dogs (LOAD) and Canine Brief Pain Inventory (CBPI) assessments at baseline, before and after the first 5km run, and before and after the final 16km run. On the LOAD questionnaire, undenatured type II collagen group had improved “how active is the dog” (P=0.03) and less “stiffness after a lie down” (P=0.041) compared with placebo at pre 5km. Undenatured type II collagen appeared to mitigate the development of pain after exercise compared with placebo, as related to the CPBI assessment. Undenatured type II collagen dogs had lower “pain at worst” pre 5km (P=0.021), “pain at least” post 5km (P=0.015), “pain at average” post 5km (P=0.046), and “pain as it is now” post 16km (P=0.006) compared with placebo dogs. Undenatured type II collagen was more effective than placebo at mitigating inflammation on gait analysis per the FRK Inflammation Index. Undenatured type II collagen dogs had a 6.42 lower FRK Inflammation Index score at 24h post 5km (P=0.032) and 6.3 lower score at 24h post 16km (P=0.029), indicating the mitigation of inflammation on gait analysis. When considering the change between timepoints, undenatured type II collagen had a lower increase in FRK Inflammation scores compared with placebo for baseline to pre 5km (p<0.001), pre 16km to 24h post 16km (P=0.028), and pre 16km to 48h post 16km (P=0.027). Undenatured type II collagen supplemented Labrador Retrievers improved pain assessment variables and improved FRK Inflammation Index on gait analysis.