The use of surfactants in washing crude oil contaminated soil is a new area of application and several approaches have been proposed for treating petroleum hydrocarbon contaminated sites. Surfactant applications in enhancing ex-situ soil-washing have been studied. These studies indicated that the removal of oil using surfactant solutions occurs at concentrations above and below the surfactant’s critical micelle concentration (CMC). The present work studied the removal of crude oil from sand using biosurfactants produced from agro-industrial wastes by Pseudomonas aeruginosa. The microorganism producer was the strain P. aeruginosa L2-1. This strain was able to produce rhamnolipids from cassava wastewater added with waste cooking oil showing excellent properties as low surface tension (30 mN m^-1) and low CMC (26.5 mg l^-1 ). The control (distilled water) washing showed an oil removal of 20%. The crude oil removal increased as the concentration of rhamnolipid solutions increased. The total of crude oil removal attained by rhamnolipid solution at CMC concentration was of 69%. Increasing the concentration of rhamnolipid, the crude oil removal reached 71% at 0.1% of rhamnolipid (w/v), 79% at 1% rhamnolipid (w/v) and 83% at 5% rhamnolipid (w/v). These results showed the potential use of the product in oils spill accidents contributing also to a new perspective of use of agro-industrial wastes.