The influence of physical activity on metabolic health in overweight dogs is unknown. This study was conducted to evaluate biomarkers of immunometabolic health in relation to changes in physical activity and adiposity. Client-owned overweight dogs participated in a 12-wk intervention based on caloric restriction combined with a training program (fitness and diet [FD] group, n = 8), or caloric restriction alone (diet-only [DO] group, n = 8). Physical activity was monitored by accelerometry. All dogs were fed the same diet and achieved similar weight loss. Fasting blood samples were collected before and after 6- and 12-wk intervention. Insulin resistance was evaluated from plasma insulin and C-peptide as well as homeostasis model assessment. Inflammation and dyslipidemia were evaluated from circulating leptin, adiponectin, C-reactive protein (CRP), monocyte chemoattractant factor-1 (MCP-1), interleukin-8 (IL-8), and cholesterol. Accelerometer counts in both groups were high compared with previous reports of physical activity in overweight dogs. No difference in blood parameters was evident between groups, evaluated by linear mixed-effects model (P > 0.05). Within the groups, the following changes were significant by t-test (P < 0.05): leptin decreased in both groups. Within the FD group, IL-8, MCP-1, and CRP decreased at 6 wk and IL-8 and cholesterol at 12 wk. Within the DO group, C-peptide and HOMA decreased at 6 wk and C-peptide at 12 wk. We conclude that, for both groups, weight loss resulted in minor indications of improved immunometabolic health, whereas this level of physical activity did not add further benefits.