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Nickel is an ideal non-noble metal anode catalyst for direct urea fuel cell (DUFC) due to its high activity. However, there exists a large over potential toward urea electro oxidation. Herein, NiCo/C bimetallic nanoparticles were prepared with various Co contents (0,10,20,30 and 40wt%) to improve the activity. The best Co ratio was 10% in the aspect of cell performance, with a maximum power density of 1.57 mWcm-2 when 0.33M urea was used as fuel, O2 as oxidant at 60oC. The effects of temperature and urea concentration on DUFC performance were investigated. Besides, direct urine fuel cell reaches a maximum power density of 0.19 mWcm-2 with an open circuit voltage of 0.38V at 60oC.