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Noise exposure may cause auditory and extra-auditory effects. School teachers and students are exposed to high noise levels which have an impact on perceptual-cognitive and neurobehavioral aspects. The latter influence teaching conditions and student school performance. A Protocol was defined and parameters to be investigated were identified for acoustic characterization of unoccupied and occupied school environments, assessment of users by means of questionnaires completed by teachers and students, and vocal effort evaluation. Classrooms, laboratories, auditoriums, gymnasiums, common areas, canteens and outdoor areas were analysed in terms of acoustic features and identification of the origin of noise. The Protocol was tested in three kindergartens, three primary schools and three secondary schools placed in Rome, Florence and Perugia. Results of nine case studies are presented, including comparisons of objective and subjective investigations. Generally, the acoustic performances of the spaces under investigation do not meet the requirements of current Italian legislation. In particular, student activity determines high noise levels in laboratories, gymnasiums, and canteens. Students notice that noise mainly causes loss of concentration, fatigue, boredom, and headache. The outcomes of this research will be the starting point to define strategies and solutions for noise control and mitigation in schools and to draft guidelines for the acoustical school design.