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Maternal factors such as fear and stress influence offspring welfare in many species, but this area of research has been underexplored in dogs. As a large proportion of society’s dogs originate from commercial-breeding (CB) kennels, further investigation in this environment is crucial to ensure canine welfare. The current study aimed to refine a set of welfare metrics used to explore associations between maternal factors and litter welfare in CB kennels. Dams (n = 28) from five USDA-licensed CB kennels were tested at 6 and 1-week prepartum, and 4 and 8-weeks postpartum using a stranger approach test, physical health assessment, exposure to novel stimuli, social isolation test, hair cortisol concentration, fecal glucocorticoid metabolites (FGM) and fecal immunoglobulin A (IgA) concentrations. Puppies (n = 138) were tested at 4 (4W) and 8-weeks (8W) old using equivalent measures. Preliminary analyses of the metrics collected during puppy social isolation revealed that locomotion was positively correlated with exploration (4W: rs=0.46, P < 0.001; 8W: rs=0.39, P < 0.001), escape attempt duration (4W: rs=0.26, P = 0.002; 8W: rs=0.31, P < 0.001), frequency of stress behaviors (4W: rs=0.22, P = 0.01; 8W: rs=0.40, P < 0.001), vocalization (4W: rs=0.18, p=0.04; 8W: rs=0.20, P = 0.02), and fecal IgA (4W: rs=0.24, P = 0.05). Stationary behavior was negatively correlated with escape attempt duration (4W: rs=-0.37, P < 0.001; 8W: rs=-0.43, P < 0.001), frequency of stress behaviors (4W: rs=-0.22, P = 0.01; 8W: rs=-0.32, P < 0.001), and physiologic metrics (4W: fecal IgA rs=-0.25, P = 0.04; 8W: FGM rs=-0.23, P = 0.01). Results suggest that, in response to social isolation, 4W and 8W old puppies exhibit one of two coping styles, proactive or passive, and the latter is associated with lower activation of the immune and stress response. The outcomes of this study will inform future research exploring maternal risk factors, including behavioral and physiologic stressors, affecting puppy developmental welfare in CB kennels.