This book constitutes the refereed proceedings of the second International Conference on Biomimetic and Biohybrid Systems, Living Machines 2013, held in London, UK, in July/August 2013. The 65 revised full papers presented were carefully reviewed and selected from various submissions. The papers are targeted at the intersection of research on novel live-like technologies inspired by scientific investigation of biological systems, biomimetics, and research that seeks to interface biological and artificial systems to create biohybrid systems.