Animal X-ing

• Brewster: A male pigeon who owns "The Roost", a cafè in the basement of your town's museum. Once the player buys coffee on a regular basis, Brewster warms up and becomes chattier.
• Celeste: Female owl with pink bow who works in the observatory. She is Blathers' sister, and allows the player to look through the telescope and design constellations. For a comical effect, the player can tell Celeste "You're cute!" If you say this, she'll be shocked and blush.
• Dr. Shrunk: Male axolotl who shows up in town occasionally. Will teach emotions for use in multiplayer chat. A wide variety of emotions exist, but the player may only know four at a time.
• Harriet: Female pink poodle who works in a salon called Shampoodle. Will style the player's hair once a day for 3,000 Bells. She is only available once Tom Nook upgrades his shop to Nookington's. If the player gets all 8 haircuts, she will offer to style the player's hair into cuts that would normally only be available to the opposite gender. The hairstyles are generated using the answers the player gives to her questions.
• Katie: A lost kitten who is seperated from her mother Kaitlyn. She appears in someone's town, while her mother appears in a different town. If players reunite her and her mother, they will receive a gift in the mail. One of these gifts is a picture of her and her mother.
• Lyle: Male weasel who sells insurance on Saturdays. Accident Insurance, which can be bought for 3,000 bells, will return 100 bells to the player for each play session the player is stung by bees or trips (caused by bad luck from either Katrina's fortune telling or wearing the King Tut Wig). Damage Insurance, for 6,000 bells, will cover fake paintings from Crazy Redd. This, as well, pays 100 bells per offense. Lyle can also book a date for Redd to visit town.
• Pascal: Male beaver who turns up on the beach occasionally. He gives out pirate-themed furniture. He will also give players either a Golden Axe or Pascal's Pic if given a scallop (which players will not find on the beach, though a different type of scallop can be found). The scallop is obtainable through a trading quest involving most NPCs.
• New villagers: In addition, there are many more villagers available now that weren't available in the previous game.

Lyle, Harriet, Ressetti, Kapp'n, Rover, Pelly, Phyllis and others can sometimes pay visits to The Roost (Harriet only appears once Nookingtons is opened).

• Blanca: A faceless cat who will visit a player's town once they have connected to the Nintendo wi-fi connection. Being faceless, Blanca is using a face someone else designed for her. She will ask the player to draw her a new face, and then will leave to go to another town.
• Blathers: Curator of the museum, Blathers accepts donations and can identify fossils and forged paintings. He is quite scared of bugs. As an owl, he tends to sleep during the day, but is awake and alert promptly at 6:00p.m.
• Booker: A guard dog who works at the gate, known as the Checkpoint. Booker handles more remedial tasks like changing the town flag and holding lost items. Though he is lazy and apathetic, Booker keeps a watch over the gate and will tell the player if anyone has recently entered the town. Both Booker and Copper now wear guardsman uniforms and wield spears (they were police officers in the GCN version).
• Copper: Another guard dog who works at the Checkpoint, Copper will let the player exit through the gate or open it for friends to visit, as well as issue the player's Friend Code.
• Crazy Redd: A sly fox who shows up in front of the Town Hall occasionally. He sells furniture for more than Nook does, but many are very rare and cannot be found at Nook's at all. He is also the sole source of paintings. Be careful when shopping here however, because he has been known to sell fakes. Redd requires a password for entry into his tent, which he will send the player in the mail every time he visits if he or she pays the initial 3,000 Bell membership fee. Non-members would have to find out the password from a neighbour and cannot buy items from him until the membership fee is paid.
• Gracie: A giraffe who drives into town occasionally. She is a famous fashion expert who gives the player rare clothing. First, her quiz must be taken, which comprises a few questions about the player's sense of fashion. she will then give the "Fashionista Badge," which is carried in the mail inventory, and an outfit which can be bought at Able Sisters. However, if she is spoken to persistently, she will offer some more clothing for a price. If she is given 5000 bells or more, she will give the player some of her designer clothing.
• Gulliver: No longer washing up on the beach as in the first game, Gulliver the seagull now flies his flying saucer, The Porpoise 5000 (a reference to the Dolphin from Pikmin, which in itself is a reference to the code name of the Gamecube while it was still in development), on the top screen. If the player shoots him down with a slingshot and finds Gulliver's five missing ship parts spread around the town, he will thank the player with an exclusive item. This is a rare occurrence, though. Unlike presents, which are seen in the sky at clocked intervals, Gulliver's ship appears at unexpected and random times. Additionally, the ship moves fairly quickly and is much harder to shoot down. A reference to Gulliver's Travels by Jonathan Swift.
• Kapp'n: A romantic kappa who drives the player to town in his cab at the beginning of the game, or when he or she moves to a different town (i.e., game card) using the "moving" feature at the civic centre.
• Katrina: A fortune-telling cat who sets up her tent outside Town Hall occasionally. She will give the player a fortune or tell his or her compatibility with a person for 100 Bells, or cleanse his or her soul (i.e., undo any fortunes given) for 10,000 Bells.
• K.K. Slider: A dog who plays guitar at The Roost on Saturday nights from 7:30 p.m. to 12:00 midnight. However, if the player talks to him before 8:00 p.m., he will tell the player to wait until 8:00 p.m., after his sound check. After his performance, he will give a bootleg of the song he just played. K.K.'s real name is actually "Totakeke", a reference to Nintendo employee Kazumi Totaka.
• Mabel: The younger of two sisters who works at the Able Sisters' tailor shop. She assists the player in making purchases or designing patterns, which can be used for many things, including clothing, caps, and accessories. Mabel's look could be a possible reference to the Sega character Sonic the Hedgehog (they are both blue hedgehogs).
• Mr. Resetti: A mole who will pop out of the ground and scream a lengthy lecture at the player if he or she does not save before quitting. When the player turns the power off without saving, then begins to play again, he or she is not allowed to save or go back inside the house without first taking a couple steps forward and having him yell at them. After a couple times of this, he will begin to ask the player to answer his questions by either typing it in the keyboard, choosing between one of the two options, or selecting a degree of regret on a slider. He is very annoying, and often makes the player regret his or her mistake. His final remark is often about the player's hygiene.
• Pelly: The cheery pelican who works at the Town Hall during the day, handling both Civic Centre and Post Office duties. Pelly has feelings for Pete, but he doesn't love her.
• Pete: A hardworking pelican who works for the Post Office, delivering mail at 9:00a.m. and 5:00p.m. He can be shot out of the sky using a slingshot. Pete is dating Phyllis, who works in the Town Hall. It is apparently a rocky relationship.
• Phyllis: Pelly's sister, the grumpy purple pelican who works at the Town Hall during the night, handling both Civic Centre and Post Office duties. She is dating Pete, the mail-carrier. She is apparently somewhat abrasive to him.
• Rover: A blue cat who runs Tag Mode. He appeared in the introductory sequence of the original Animal Crossing. He shows up in the Roost occasionally, and mentions his old life of hopping the train, travelling, and his encounters with Blanca.
• Sable: The older sister at the Able Sisters' tailor shop. Her job seems to consist entirely of sewing. Though she is shy at first, she will warm up to the player if they spend enough time speaking with her. Occasionally she hints at a lifetime, unrequited crush on Tom Nook.
• Saharah: A camel who delivers rare carpets and wallpapers. If the player helps with her deliveries, she will reward the player with either a carpet or a wallpaper. The player is allowed to choose either the carpet or the wallpaper; however, they do not know what design it is until they receive it.
• Snowman: Appears when the player builds him in the winter. If he is constructed well, he will send the player a piece of rare furniture in the mail. If he is created out of proportion, he complains about his life and pities his own existence.
• Sow Joan: An old boar who has been selling turnips for over 60 years. She will sell the player white turnips to play the "stalk market," or red turnips to plant and grow to sell for an enormous profit.
• Timmy & Tommy Nook: Tom Nook's nephews who manage the top floor of Nookington's. They attempt to speak in unison, but Tommy always lags behind.
• Tom Nook: A male tanuki and owner of the town's main general store. He sells the player a house when they start the game, and offers to expand it whenever the player pays off their mortgage entirely. This can happen a maximum of six times, when the house has reached its maximum size. Nook will occasionally get emotional, and if asked will discuss his failed dreams of a shop in the big city. In the American/PAL release, Nook is Americanized and referred to as a raccoon.
• Tortimer: Tortimer is the mayor of the town, and a tortoise (hence the name). Tortimer appears in front of Town Hall during holidays and events and during the time when the player works at Nook's Cranny. The rest of the time, he's sleeping in the back of Town Hall.
• Wendell: A blue walrus, described as a "starving artist." He now gives out patterns instead of wallpaper, and which one the player gets is dependent on what the player feeds him. He considers red turnips to be a great delicacy.

Characters who do not return from the N64 and GameCube versions are Porter, Wisp, Jack, Jingle, Chip, Farley, Franklin, Don Resetti (Mr. Resetti's brother), and several villagers from the last game.