<P><B>Abstract</B></P> <P>Hydrogen production from cassava starch processing wastewater by two-stage thermophilic dark fermentation and microbial electrolysis was investigated. Single-chamber membrane-free microbial electrolysis cell with applied voltage of 0.6 V was optimum for hydrogen production with hydrogen yield of 245 ml H<SUB>2</SUB> gCOD<SUP>−1</SUP>. Continuous microbial electrolysis reactor has a maximum hydrogen yield of 182 ml H<SUB>2</SUB> gCOD<SUP>−1</SUP> at HRT of 48 h with energy recovery efficiency of 217%. The continuous integrated two-stage dark fermentation and microbial electrolysis have hydrogen yield of 465 ml H<SUB>2</SUB> gCOD<SUP>−1</SUP> with 2 times hydrogen yield improving compared with a single stage and a maximum COD removal of 58% was achieved. Dominated bacteria at an anode of microbial electrolysis cell were exoelectrogens belong to <I>Brevibacillus</I> sp. <I>Caloranaerobacter</I> sp. and <I>Geobacillus</I> sp. Continuous two-stage thermophilic dark hydrogen fermentation and microbial electrolysis were efficient processes for hydrogen production from cassava starch processing wastewater.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Integrated two-stage dark fermentation and microbial electrolysis was investigated. </LI> <LI> Applied voltage of 0.6 was optimum for microbial electrolysis cell. </LI> <LI> Hydrogen yield of continuous single stage MEC was 182 ml gCOD<SUP>−1</SUP>. </LI> <LI> Hydrogen yield of continuous two-stage fermentation and MEC was 465 ml gCOD<SUP>−1</SUP>. </LI> <LI> <I>Brevibacillus</I> sp. <I>Caloranaerobacter</I> sp. and <I>Geobacillus</I> sp. were dominated in MEC. </LI> </UL> </P>