This book constitutes the refereed proceedings of the 16th Italian Workshop on Artificial Life and Evolutionary Computation, WIVACE 2022, held in Gaeta, Italy, during September 14–16, 2022.  The 21 full papers and 3 short papers included in this book were carefully reviewed and selected from 45 submissions. They were organized in topical sections as follows: answer set programming; networks and complex systems, metaheuristics, robotics, and machine learning Chapters 7, 8, and 9 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.