The Federal Court on Thursday dismissed the appeal, with Justices Arthur Emmett, Jayne Jagot and John Nicholas concluding "there was an infringement in respect of the recording Down Under".
EMI filed the appeal in February last year after it was found the famous flute riff from the band's smash hit was unmistakably the same as children's tune Kookaburra Sits in the Old Gum Tree.
The decision left Kookaburra's copyright owners, Larrikin Music, poised to claim millions of dollars in unpaid royalties from EMI, and Down Under songwriters Colin Hay and Ron Strykert.
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EMI listed 14 grounds for appeal and sought orders that Hay and Strykert did not breach copyright with their work.
It argued the inclusion of two bars from Kookaburra was at most a form of tribute to the piece, penned by Toorak teacher Marion Sinclair more than 75 years ago for a girl guides competition.
The company also said the trial judge had placed insufficient weight on the fact that the similarities between the two songs had gone unnoticed for many years.
EMI was ordered to pay Larrikin's costs.
