Anthropogenic noise is a growing ubiquitous and pervasive pollutant as well as a recognised stressor that spreads throughout natural ecosystems. However, there is still an urgent need for the assessment of noise impact on natural ecosystems. This article presents a multidisciplinary study which made it possible to isolate noise due to road traffic to evaluate it as a major driver of detrimental effects on wildlife populations. A new indicator has been defined: AcED (the acoustic escape distance) and faecal cortisol metabolites (FCM) were extracted from roe deer faecal samples as a validated indicator of physiological stress in animals moving around in two low-traffic roads that cross a National Park in Spain. Two key findings turned out to be relevant in this study: (i) road identity (i.e. road type defined by traffic volume and average speed) and AcED were the variables that best explained the FCM values observed in roe deer, and (ii) FCM concentration was positively related to increasing traffic volume (road type) and AcED values. Our results suggest that FCM analysis and noise mapping have shown themselves to be useful tools in multidisciplinary approaches and environmental monitoring. Furthermore, our findings aroused the suspicion that low-traffic roads (< 1000 vehicles per day) could be capable of causing higher habitat degradation than has been deemed until now. Palabras clave: Disturbance, Noise, Wild boar