Raw glycerol, byproduct from bio-diesel production process, is used as carbon substrate in several biotechnological applications. Using Clostridium butyricum F2b, 47.1 gL(-1) of 1,3-propanediol was produced in batch anaerobic cultures while substrate uptake rate (r(s), expressed in g L-1 h(-1)) increased with increase in glycerol concentration in the medium. In continuous cultures, microbial behaviour was studied in transitory states after addition of 1,3-propanediol in the chemostat vessel. Microbial growth was not affected by the high 1,3-propanediol (which was added in the chemostat vessel) concentration, while butyric and acetic acids concentrations were increased. In a two-stage continuous culture, 43.5gL(-1) of 1,3-propanediol was produced with a total volumetric productivity of 1.33gL(-1)h(-1). Yarrowia lipolytica ACA-DC 50109 was grown in nitrogen-limited aerobic cultures on raw glycerol and it exhibited remarkable biomass production even at high giycerol concentration media, while rs decreased with increase in glycerol concentration. Citric acid was produced after nitrogen depletion in the medium, with the highest quantity of 62.5 g L-1, and yield on glycerol consumed was 0.56 g g(-1). Fatty acid analysis of total cellular lipids showed that glycerol concentration increase in the growth medium somehow increased the cellular unsaturated fatty acids content of lipids. Mortierella isabellina ATHUM 2935 exhibited satisfactory growth in nitrogen-limited aerobic cultures with raw glycerol used as sole substrate. When high initial glycerol quantities were employed (e.g. 100gL(-1)), 4.4gL(-1) of lipid were accumulated corresponding to around 51% (wt/wt) of lipid in dry weight. rs constantly decreased with increase in glycerol concentration in the medium, and in all cases notable glycerol quantities remained unconsumed in the medium. (c) 2007 Elsevier Ltd. All rights reserved.