The achievement "Principessa in Another Castello" is a reference to Super Mario Bros., where Toad says "Thank you Mario! But our princess is in an another castle!"

The achievement "Grand Theft Dressage" is a reference to the Grand Theft Auto series.

The game's soundtrack was included as one of the pre-order bonuses for the game. However, the header information in the music files indicates that they were encoded by "arsa13", a user responsible for putting up illegal copies of the soundtrack on torrent sites.

This suggests that Ubisoft had downloaded a pirated version of their own soundtrack to publish as a pre-order incentive.

Although Ezio had his left ring finger branded in the second game, the brand cannot be seen in Brotherhood and never reappears.