Measurement of heart rate (HR) has been used to determine the stress response and evaluate well-being in livestock species since the early 1970s. However, according to the latest physiological and behavioural studies, heart rate variability (HRV) has been shown to be more accurate for studying the activity of the autonomic nervous system. Alpacas are domestic camelids that are used to obtain fibre, in animal therapies and as a companion animal. Numerous herds are found in the central area of Chile and many specimens have been exported to Europe, North America, Australia and China, illustrating the global interest in this type of animal native to the southern cone of America. Although studies have been carried out that describe the stress response and its severity in domestic camelids to handling, studies in these species are scarcer than in other species. Indeed, heart rate variability (HRV) has been barely evaluated, which according to current knowledge is the best way to evaluate the activity of the autonomic nervous system, in addition to being a non-invasive measurement. In this study, the physiological and behavioural response of alpacas to routine handling will be described and we hypothesized that stronger handling produces a greater stress response, evidenced by an increase in heart rate, lower HRV and an increase in behavioural stress responses. The response of a group of alpacas will be evaluated to social stressors such as interaction with humans and social isolation, and to handling such as physical restraint for routine handling (e.g. claw trimming). The measurement will be performed with a Polar Vantage V® cardiac monitor. With this evaluation and better understanding the stress response to handling, practical recommendations can be made to reduce stress and promote better welfare for alpacas.