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This book constitutes the thoroughly refereed proceedings of the 5th International Workshop on Hybrid Systems Biology, HSB 2016, held in Grenoble, France, in October 2016.The 11 full papers presented in this book were carefully reviewed and selected from 26 submissions. They were organized and presented in 4 thematic sessions also reflected in this book: model simulation; model analysis; discrete and network modelling; stochastic modelling for biological systems.