Lichens are one of the most responsive components of the ecosystem to reactive forms of nitrogen. In this work, we selected the lichen genera Cladonia and Usnea, composed of terricolous and epiphytic lichens respectively, and described as sensitive to nitrogen, to test the effects of different doses of nitrogen on lichen physiological parameters (photobiont and mycobiont vitality, chitin quantification, nitrogen content and stable isotopes analysis). The main objectives were to check if the activation of protective mechanisms could be stimulated in case of chronic stress (low nitrogen increase for prolonged time), and, if so, if a toxicity threshold could be identified above which these mechanisms fail. The two lichen genera were generally affected by prolonged exposure to increased nitrogen availability. However, Cladonia rangiformis was able to maintain physiological functioning at the lowest nitrogen doses used, whereas thalli of Usnea become overwhelmed. Moreover, the mycobiont appeared to be more sensitive than the photobiont responding to lower nitrogen doses. Although only studies of longer duration and testing more nitrogen doses will be able to determine an accurate toxicity threshold, these results give important clues on the use of lichens as biomonitors for the establishment of environmental policies. Keywords: Chitin, Cladonia, Ecophysiology, Isotopic signature, Toxicity threshold, Usnea.