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Simple Summary: Puppies from commercial breeding kennels (CBKs) are transported by ground from their facilities of origin to a distributor at approximately 8 weeks of age, which is a sensitive period in dogs' early development. Experiencing high levels of fear and stress during this transition potentially jeopardizes their short- and long-term welfare. However, no research has explored the effect of transportation on puppy welfare. Therefore, we tested 383 puppies at 12 CBKs and again approximately 48 h after transportation to a distributor. We measured puppies' behaviors in isolation and during a stranger-approach test, and conducted a physical health assessment. Feces were also collected from each litter and from one focal puppy per litter for insight into their stress response, immune function, and presence of intestinal parasites. Overall, puppies were physically healthy before and after transportation. However, behavioral and physiological findings suggest puppies experienced an increase in distress after transportation. Results also suggest that puppies may have solicited more contact from an unfamiliar person as a coping strategy following an acute stressor after transportation. Future studies should investigate risk factors associated with transportation protocols and identify interventions to help optimize puppy welfare during this transition. Many puppies from commercial breeding kennels (CBKs) are transported by ground from their kennels of origin to a distributor. This experience may elicit fear and stress during a sensitive developmental period, which may in turn negatively impact the puppies' short- and long-term welfare. This study aimed to measure short-term effects of transportation on puppy welfare metrics. Eight-week-old puppies (n = 383) from 12 CBKs were tested at their kennels (pre-trans) and ~48 h after arriving at a distributor (post-trans). At each location, puppies underwent an isolation test, a stranger-approach test, and a physical health assessment. Behavioral responses to testing were scored from videos. Fecal glucocorticoid metabolites (FGM), fecal secretory immunoglobulin A (sIgA), and presence of intestinal parasites were also analyzed. Linear mixed-effects models identified decreased exploration (p < 0.001), and increased locomotion (p < 0.001) and escape attempts (p = 0.001) during the post-trans isolation test. Increased affiliative behavior (p < 0.001), FGM (p < 0.001) and sIgA (p = 0.014) were also observed post-trans. Findings support good physical health both pre- and post-trans, while behavioral and physiological changes suggest increased puppy distress post-trans. Higher post-transport affiliative behavior may indicate that puppies sought social support as a coping strategy after experiencing transport-related distress. Future studies should explore the efficacy of transportation-related interventions to mitigate puppy distress. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]