The combined pressures of anthropogenic activities require rapid assessments of environmental impacts on ecosystems to develop strategies for sustainable management and biodiversity conservation. This study investigates the consequences of the anthropogenic effects in the upper Val d'Agri (Basilicata, Italy) by employing a multi-taxonomic analysis to investigate spatial dynamics and species responses to these pressures. By cataloguing 151 diverse species of small mammals, reptiles, birds, ground beetles, and lichens, we established a valuable baseline for analysing the local biodiversity. We assessed the relationship between anthropic impacts (industrial areas, roads, agriculture, forestry, etc.), vegetation cover, and species diversity. Our findings revealed a negative relationship between the number of species and proximity to industrial areas, emphasising the impact of these activities. Moreover, we observed a predominance of generalist biological traits across taxa in all the study sites rather than an increase in species specialisation with increased distance. This may suggest a homogenising effect caused by various anthropogenic activities that cause habitat fragmentation and species mortality, underlining the lasting influence of these activities on the biodiversity of the upper Agri Valley. Additionally, our study identifies numerous protected species, highlighting their vulnerability and emphasising the necessity for targeted conservation efforts to safeguard their existence.