This book constitutes the refereed proceedings of the Second International Symposium on Artificial Life and Intelligent Agents, ALIA 2016, held in Birmingham, UK, in June 2016. The 8 revised full papers and three revised short papers presented together with two demo papers were carefully reviewed and selected from 25 submissions. The papers are organized in topical sections on modelling; robotics; bio-inspired problem solving; human-like systems; applications and games.