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Assassin's Creed II takes place in an open world with nonlinear gameplay, allowing the player to roam freely within several regions throughout late fifteenth-century Italy such as Venice, Florence, Forlì, San Gimignano, and the Tuscan countryside. The Animus 2.0, a new version of the machine of the same name present in Assassin's Creed, provides in-game context for changes and additions to several game elements. A database is also available, providing extra historical information about key landmarks, characters and services that the player encounters. The health system has been made more dynamic, with synchronization to the Animus and causing the character to recover only from minor injuries. More grievous injuries require visiting a street-side doctor or use of medicine (which can be purchased from doctors or found on bodies).
The player may now swim in water, and Eagle Vision—the ability to identify specific people and landmarks—can now be used in third-person view and while moving. A young Leonardo da Vinci is present in the game, aiding the player by creating new weapons from translated "Codex pages" that Altaïr, the original game's main character, left behind for future Assassins' analysis and insight. Within the game, the player will be able to use Leonardo's flying machine (based on real-life plans by Leonardo) during one mission. The player also has the ability to control a carriage in one level, and can row gondolas, as well as ride horses at any point in the game where they are readily available between towns and cities. The setting of the various places the player may go to have been made more detailed and in-depth; civilians can carry objects and sometimes cough or sneeze. Additionally, the player can hire different groups of NPCs, such as mercenaries, courtesans, or thieves; these groups can be used to fight, distract, or lure guards, respectively. A day and night cycle has been added to the game, giving the game more of a sense of time, in addition to setting missions and events at certain times of the day.
There are many ways to interact with NPCs. Money thrown to the ground, or a corpse carried and then deposited on the ground, may also serve as a distraction for both guards and peasants. There are also several different types of enemies, some more agile or stronger than others, and some of which will actively search hiding places where Ezio was last seen.
The combat system is more complex than that of its predecessor, with the ability to disarm opponents using counter-attacks while unarmed. If the player steals an enemy's weapon, it is possible to follow up with an attack that instantly kills the enemy. Da Vinci provides the player with specialized weaponry, such as the dual hidden blades, poison blade and the miniature wheellock firearm, which are all based on schematics found in Altaïr's Codex pages. Generic swords, cutlasses, maces, axes and daggers can all be purchased from vendors in each city or otherwise looted from corpses (some items, such as spears and brooms, cannot be acquired from vendors). In addition, players are able to purchase artwork for their villa, obtain new armor as the game progresses, and even dye Ezio's clothing with a number of different colors. Other equipment includes larger pouches to carry more throwing knives and medicine. Six additional weapons can be unlocked by connecting a PSP with Assassin's Creed: Bloodlines to the PS3.
The Auditore family's countryside villa, located in Monteriggioni, acts as Ezio's headquarters: the surrounding property can be upgraded, drawing income for the player's use. There are several outlets for using currency, with vendors selling items such as medicine, poison, weapons, repairs, upgrades, paintings, and clothing dyes. When these shops are renovated, Ezio receives discounts at the shops on the goods they sell. Purchasing weaponry, armour sets and artwork also contributes to increasing the villa's overall worth, in turn generating more income for Monteriggioni.
There is now a broader array of methods for hiding or blending in the area. One can dive underwater to break guards' line of sight, and blending may be performed with any group of people, rather than only a specific type (as in the first Assassin's Creed). The game features a notoriety system, with guards more alert to Ezio's presence depending on his behavior, location, and current mission. This infamy can be reduced through bribery, removing wanted posters, or assassinating corrupt officials.