Photofermentation seems to be an attractive mode of generating biohydrogen from fermentation effluent. Use of succinate fermentation effluent, however, has not been reported. Rhodobacter sphaeroides KKU-PS1 and Rhodopseudomonas palustris were acclimatised in succinate. It was determined that the KKU-PS1 was superior with respect to hydrogen productivity and was selected for further experiments. Photofermentation in succinate by the KKU-PS1 validated, generating 1217 mL H-2/L of cumulative hydrogen at a maximum rate of 6.7 mL H-2/L/h. Photofermentation from each single carbon sources that are components of effluent was performed and it was determined that acetate and succinate promoted the fastest growth of KKU-PS1 and hydrogen evolution, respectively. Photofermentation by the strain using mixed substrates mimicking diluted bio-succinate effluent produced yielded 1005 mL H-2/L cumulative hydrogen at a maximum rate of 4.1 mL H-2/L/h. The study highlighted potential of utilizing bio-succinate fermentation effluent for biohydrogen production, with further optimization required. (C) 2019 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.