Director Carlos Saldanha became interested in casting singer Bruno Mars in his first voice-over role, after seeing him on Saturday Night Live (1975). After working with Mars, he re-wrote his character to make him less "macho", and, after a recording session, had a segment reanimated to fit his singing.

There are roughly 150 Spix's Macaws that make up the giant "2" of the U.S. teaser poster. The number of Spix's Macaws within that formation seems to loosely follow the real-life population of the species left in existence (most of which are kept in captivity around other parts of the world).

During the "animal auditions" scene, one talent involves a monkey swinging on a rock, tied to a vine. A reference to the notable "Wrecking Ball" music video of singer Miley Cyrus.

The boat the characters take up the Amazon River is called "Céu Azul", a translation of Blue Sky, name of the production company.

Although Eva is portrayed to be a terrible off-key singer, Bebel Gilberto, who voices the character, is actually a very good singer and accomplished musician.

This is Don Rhymer's last film as a writer, before his tragic passing.

As of 2014, Ледниковый период (2002) and Рио (2011) are the first Blue Sky Studios films to have spawned sequels.

The anteater Charlie was modeled and named after Charles Chaplin, notably for his mute mannerisms and signature bowler hat.

In an earlier written storyboard concept, Fernando, the helpful street urchin from the first Рио (2011), was shown to have been adopted as the son of Linda and Tulio, and would accompany them on their jungle journey, in the process showing his newly specialized skills in field work. Due to pacing reasons, his role was cut down to a cameo at the beginning, where he has clearly aged since the first movie.

Janelle Monáe's participation in the film, owes to a meeting between the singer and Carlos Saldanha at her Rock in Rio (1985) performance in September 2011.

In Рио (2011), after Blu and Jewel escape from the smugglers, Blu can be heard saying "Have you ever heard 'It's a jungle out here'?" In Rio 2, one of the movie's themes is called "It's a Jungle out Here."

When Carla and Bia are channel surfing, there are clips from previous Blue Sky Studio films shown briefly prior to Linda and Tulio's news report: Ледниковый период 4: Континентальный дрейф (2012) and Эпик (2013).

At the talent competition, the mosquitoes perform a small extract from Wolfgang Amadeus Mozart's Rondo Alla Turca.

The mustachioed monkey that the chief tree thief carries around, is called an emperor tamarin after German monarch Kaiser Wilhelm II, who bore an amazing resemblance to the creatures.

In Serbia, Felipe is dubbed by a famous rapper Ajs Nigrutin, and the evil boss is voiced by football player Savo Milosevic.

During the confrontation with the lumberjacks in the forest, one of them asks Linda (Leslie Mann) and Tulio (Rodrigo Santoro) if they are lost, to which Tulio replies "Lost? Lost? No, we are on our honeymoon." Santoro played a supporting character in the series Остаться в живых (2004), where he was also stranded with his girlfriend on a forest-covered island.

In Rio, when Blu and Jewel meet for the first time, she starts speaking rapidly in Portuguese. But in Rio 2, when she meets her father, they start speaking English right away, even though her father doesn't know that she speaks English to her family.

The little capibara at the audition scene who gets swallowed by the big black jaguar, is singing the song "Memory", which, appropriately, comes from the musical "Cats".

The Brazilian newspaper that the Big Boss throws down on the table at the logging camps reads: Rare Birds in the Amazon - Latest news from Dr. Tulio Monteiro, Ornithologist.

Blue Sky Studios' second film to be rated G. The first being Dr. Seuss' Horton Hears a Who! (2008), (not counting Rio (2011) which is rated PG).

The fourth Blue Sky Studios film to be produced in 2.35:1, after Рио (2011), Ледниковый период 4: Континентальный дрейф (2012) and Эпик (2013).

This is the last Blue Sky Studios film to be composed by John Powell until Ferdinand (2017) released 3 years later.

Josh Brolin were considered for the role of Eduardo.

Isla Fisher were considered for the role of Gabi.

Jason Bateman were considered for the role of Roberto.

Ian McShane were considered for the role of Eduardo.

Riz Ahmed were considered for the role of Roberto.

Taron Egerton were considered for the role of Roberto.

Nigel is first seen reciting passages from William Shakespeare's Hamlet. His "death" at the end involves an attempt to kill a dueling participant via a poison which is ultimately administered to the wrong person, as also happens at the end of Hamlet.

George Lopez and Andy Garcia previously starred together in Крошка из Беверли-Хиллз (2008), respectively.