Human activities represent one of the most dramatic forms of environmental change, affecting global biodiversity. Anthropogenic noise has increased in the last century with detrimental effects for wildlife on both individual and population levels. Sound pollution can compromise animal heath by imposing chronic stress and damaging the auditory system, affect intra- and inter specific relationships by disrupting communication and masking the presence of predators. Shrews produce high-pitched laryngeal calls (“twittering calls”), with unclear function. It has been suggested as rudimentary echo-orientation mechanism, communication, or response to stressful stimuli. In this study, we analysed greater white-toothed shrews (Crocidura russula), trapped along a gradient of anthropogenic noise with the goal to link twittering calls to exposure to environmental stressors, through the analysis of faecal glucocorticoid levels. We hypothesize that animals inhabiting noisier locations would experience higher levels of stress, produce more twittering calls, and reduce exploratory behaviour to balance stress demands. We live trapped 25 shrews in 6 locations with different sound patterns and maintained them in captivity for 5 consecutive days, for repeated measurement of twittering calls, exploratory behaviour, and stress levels. Results show that variation in noise levels influences the shrews’ stress response, independently of the average noise levels. Neither average noise levels nor its variation induced a change of vocalisations. While time in captivity resulted in a reduction of exploratory behaviour and twittering calls, we did not find evidence supporting a link between stress levels and twittering calls.