Biohydrogen (bioH(2)) production via dark fermentation is an attractive approach to overcome the drawbacks of conventional hydrogen production methods and represents a preliminary alternative for the management of organic wastes. Fundamental studies are still required to enhance the performance of bioH(2) production systems, with emphasis on the development of novel reactor configurations. The anaerobic structured-bed reactor (ASTBR) is a recently developed configuration with great potential for bioH2 production, although operating strategies are still required to minimize biomass washout in such systems. In this context, calcium dosing has been investigated as a strategy to enhance both biomass retention and bioH(2) production rates in the ASTBR. The present study employed varying COD/calcium ratios (4423, 2079, 1357, 1012, 884, and 632) in continuous experiments under mesophilic conditions (25 degrees C). Calcium dosing effectively enhanced biomass retention within the ASTBR, directly increasing the availability of metabolic energy for different metabolic pathways rather than cell synthesis. An optimal COD/calcium ratio of 1360 was mathematically estimated for bioH(2) production, which is consistent with the results obtained experimentally. The specific organic loading rate (SOLR) was better controlled at this ratio, indicating the establishment of balanced conditions in terms of substrate availability and biomass concentration. Conversely, bioH(2) production was severely impaired at COD/calcium values below and above the optimal range, most likely due to enhancement of the homoacetogenic pathway as a result of unbalanced conditions in the SOLR. Furthermore, biomass accumulation did not strongly affect the mean residence time of the ASTBR, facilitating its robust and enhanced solid retention. (C) 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.