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Global research is moving forward in developing biological production of hydrogen (biohydrogen) as a renewable energy source to alleviate stresses due to carbon dioxide emissions and depleting fossil fuels resource. Biohydrogen has the potential to replace current hydrogen production technologies relying heavily on fossil fuels through electricity generation. While biohydrogen research is still immature, extensive work on laboratory- and pilot-scale systems with promising prospects has been reported. This work presents a review of advances in biohydrogen production focusing on production pathways, microbiology, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined.