For the period 2003 to 2009 in Brazilian municipalities of over 60,000 inhabitants, buses accounted for more than 25% of urban trips, which is a trend that is not expected to change in the medium term. Worldwide, buses rely on petroleum diesel oil as fuel. In Brazil, alternative fuels such as biodiesel, natural gas and ethanol are available, and the choice between them should depend on an evaluation of the entire life cycle of such fuels. The present study uses a Life Cycle Inventory (LCI), which is essential to the implementation of a Life Cycle Analysis (LCA), to analyze 6 energy alternatives: petroleum diesel oil (D100), pure biodiesel (B100), biodiesel mixed with petroleum diesel (B5), compressed natural gas (CNG), CNG in conjunction with B5 (B5CNG) and additive hydrous ethanol (E95). In saving total energy consumption, D100, B5 and B5CNG stand out, in that order. If the use of renewable energy and the reduction of net CO2 emissions are the goals, E95 and B100 should be given preference.