Abstract
Hydrogen is being increasingly recognized as a promising clean, renewable energy carrier. Among the available production pathways, biological processes, particularly dark fermentation of residual biomasses and agricultural by-products, represent an appealing approach aligned with circular economy principles. These feedstocks are abundant and low cost; however, their relatively low energy density constrains process efficiency. To mitigate this limitation, research efforts have concentrated on optimizing substrate composition and implementing pre-treatment strategies to enhance hydrogen yields. Numerous studies have explored the potential of agricultural and livestock residue, yet reported outcomes are often heterogeneous in terms of units, systems, and experimental conditions, complicating direct comparison. This review consolidates current knowledge and identifies effective strategies to optimize biohydrogen generation. Among the investigated substrates, corn stover emerges as the most promising, with hydrogen yields up to 200 [mL H2/gVS (Volatile Solids)]. Evidence further suggests that inoculum processing, including enrichment or pre-treatment, can substantially improve performance, often more effectively than substrate processing alone. When both inoculum and substrate are treated, hydrogen yields may increase up to fourfold relative to untreated systems. Overall, integrating suitable feedstocks with targeted processing strategies is crucial to advancing sustainable biohydrogen production.