Ph.D., Faculty of Science, University of the Wiwatersrand, 2011 / Biological production of hydrogen gas has received increasing interest from the international community during the last decade. Most studies on biological fermentative hydrogen production from carbohydrates using mixed cultures have been conducted in conventional continuous stirred tank reactors (CSTR) under mesophilic conditions. Investigations on hydrogen production in reactor systems with attached or self-immobilized microbial growth have also appeared recently in the literature. These investigations on attached or self-immobilised bacteria involve hydrogen production in the mesophilic and thermophilic temperature range. The present study investigated the design and operational features of anaerobic fluidized granular bed bioreactor (AFGB) system which would facilitate the simultaneous achievement of high productivities (HPs) and high hydrogen yields (HYs).Where high HPs is greater than 120 mmol H2 /(L.h) and HYs greater than 4 mol H2/mol glucose. Theoretical maximum yield for an exponentially growing non-granulated bacterial monoculture will always be less than the thermodynamic maximum of 4 mol H2 /mol glucose: C6H12O6 +4H2O → 2CH3COO- + 4H2 + 4H+ + 2HCO3. The design features included reducing the total non-working or dead volume of bioreactor system. The operational improvements included application of thermophilic temperatures and high rates of de-gassed effluent recycling through the fluidized granular bed. An example of an optimal ratio of effluent recycle rate (R) to bioreactor working volume (V) was (3.0 L/min)/(3.2 L/min) = 0.94 minutes. Under conditions where temperatures were maximised and V/R were minimized the HPs increased to 21.58 L H2 /h. Also under these conditions the HYs increased above 3.0 mol H2/mol glucose. Specific hydrogen productivity for the fluidized granular bed increased from 0.25 L H2 / (g BM.h) or 8.83 mmol H2 / (g BM.h) at 45 oC to 0.525 L H2 / (g BM.h) or 18.03 mmol H2 / ( g BM.h) at 70 oC. A 3.64 fold increase in hydrogen yield occurred with an increase in temperature from 45 oC to 70 oC. XX When expressed in terms of glucose, this represents an increase from 1.34 mol H2 /mol glucose to 4.65 mol H2 /mol glucose. Finally, an evaluation of the net energy production by the AFGB system revealed a positive energy balance, making thermophilic biohydrogen production energetically viable from a commercial perspective.