Pt-Ru and Pt-Re and Pt-Ru-Re nanoparticles supported on functionalized multi-walled carbon nanotubes (f-MWCNT) were synthesized via modified polyol reduction method and tested thoroughly in a half cell and single direct ethanol fuel cell for ethanol electrooxidation in acidic medium. The MWCNTs were functionalized in a mixture of HNO3/H2SO4 solution for depositing a more active metal alloy nanoparticle on support material. The alloy formation of bi-metallic and tri-metallic electrocatalysts were examined by XRD analysis and more clearly explained by FE-SEM element mapping. The TEM analyses reveal that electrocatalysts nanoparticles are well dispersed on f-MWCNT, with spherical shapes and nano sizes range of 1.5-4 nm. The electrochemical analyses by cyclic voltammetry and chronoamperometry measurements reveal that tri-metallic electro-catalyst Pt-Ru-Re (1:1:0.5)/f-MWCNT exhibits the highest electrocatalytic activity and stability to- wards ethanol electrooxidation among all the synthesized electrocatalysts. The same electrocatalyst as anode in single DEFC results in excellent performance in comparison to all other synthesized electrocatalysts, with a maximum power density of 9.52 mW/cm(2) at a cell temperature of 30 degrees C. The bi-metallic Pt-Ru (1:1)/f-MWCNT and Pt-Re (1:1)/f-MWCNT produced power density of 7.48 mW/cm(2) and 4.74 mW/cm(2) at room temperature of 30 degrees C. The power density of DEFC enhanced 2.44 times, when cell operating temperature was increased from 30 degrees C to 80 degrees C using anode electrocatalyst Pt-Ru-Re (1:1:0.5)/f-MWCNT and keeping other parameters constant. The best result obtained in half cell and single DEFC using Pt-Ru-Re (1:1:0.5)/f-MWCNT electrocatalyst may be attributed to the synergistic effect of Pt, Ru and Re combined with bi-functional and ligand effects. (C) 2020 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.