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It has been recognized that oils derived from microorganism and wastewater sludge are comparable replacements of traditional biodiesel production feedstock, which is energy intensive and costly. Energy balance and greenhouse gas (GHG) emissions are essential factors to assess the feasibility of the production. This study evaluated the energy balance and GHG emissions of biodiesel production from microbial and wastewater sludge oil. The results show that energy balance and GHG emissions of biodiesel produced from microbial oil are significantly impacted by the cultivation methods and carbon source. For phototrophic microorganism (microalgae), open pond system gives 3.6 GJ higher energy gain than photo bioreactor system in per tonne biodiesel produced. For heterotrophic microorganisms, the energy balance depends on the type of carbon source. Three carbon sources including starch, cellulose, and starch industry wastewater (SIW) used in this study showed that utilization of SW as carbon source provided the most favorable energy balance. When oil extracted from municipal sludge is used for biodiesel production, the energy gain is up to 29.7 GJ per tonne biodiesel produced, which is higher than the energy gain per tonne of biodiesel produced from SIW cultivated microbes. GHG emissions study shows that biodiesel production from microbes or sludge oil is a net carbon dioxide capture process except when starch is used as raw material for microbial oil production, and the highest capture is around 40 tonnes carbon dioxide per tonne of biodiesel produced. (C) 2013 Elsevier Ltd. All rights reserved.