Despite the growing popularity of dog-assisted interventions (DAI), limited empirical evidence exists on their effect on the physiological stress response. The current systematic review examines the existing literature on the effect of a single, brief DAI on psychobiological indicators of stress in different demographics and settings, with a focus on identifying methodological strengths, weaknesses, and gaps in the field. A brief intervention is considered as lasting up to 45 minutes. Twenty-six relevant studies, each involving a group of participants receiving DAI and at least one control condition, were identified and analyzed for methodological quality and findings. The review aimed to identify gaps in knowledge and contributes to a deeper understanding of DAI, offering insights for future research. Evidence to date partially supports the notion that a brief therapeutic intervention with the assistance of a dog may influence the autonomic stress response. However, this review highlights a need to standardize methodologies when collecting psychobiological indicators of stress to clarify the relationship between DAI and physiological stress responses.