Ange Capuozzo went over for two tries from full-back to help lead Italy to the historic victory, though a much-changed Australia still had the opportunity to get themselves out of trouble in the final seconds. However, debutant Ben Donaldson sent his conversion just wide of the post with the last kick of the match.
It is Italy’s maiden triumph in 19 meetings with Australia and leaves them with two wins from two so far in the Autumn Nations Series following their success over Samoa.
However, the loss is a concern for the touring Wallabies who have now suffered back-to-back defeats after also being edged out by one point against France in Paris last weekend.
Australia trailed 17-3 at the Stadio Artemio Franchi after Pierre Bruno and Capuozzo both touched down for the Azzurri, but Tom Wright pulled one back to reduce Italy’s advantage to 17-8 at the break.
Fraser McReight scored four minutes after the break to help cut the gap further, but Capuozzo soon went over for his second try of the game after bursting through the Australian defence and touching down in the right corner.
Tom Robertson soon added a third try for the Wallabies to give them a lifeline and Cadeyrn Neville crossed at the very end of the match, but Donaldson was unable to convert as Italy held on for a famous win.
Additional reporting by PA Sport.