There are five dominant civilizations represented on the fictional TV show Babylon 5: humans, the Narn, the Centauri, the Minbari and the Vorlons; and several dozen less powerful ones. A number of the less powerful races make up the League of Non-Aligned Worlds, which assembled as a result of the Dilgar War, which occurred 30 years before the start of the series.

Languages There are three primary languages used on the Babylon 5 station: English and the fictional Centauri and Interlac. English is mentioned explicitly as the "human language of commerce,"Objects at Rest and is the baseline language of the station; written signs appear in all three languages. Other human and alien languages do exist in the Babylon 5 universe, though with the exception of Minbari, hearing them spoken is uncommon; when aliens of the same species are speaking to one another, the words heard are English, though it is presumed they are speaking their native tongue. Only in the presence of humans can the alien language be heard, to stress that the humans cannot understand what is being said.

The Gaim, pak'ma'ra, and Vorlons do not speak directly in English; in the case of the pak'ma'ra, either because they refuse to learn any language other than their own,"Learning Curve" or because they are incapable of making human sounds. Members of these races instead make use of real-time translation devices.

The principal human characters speak with an American accent, with the exception of Marcus Cole, who speaks with Jason Carter's natural English accent. Susan Ivanova, born in Russia, speaks with an American accent, as her character was raised and schooled outside Russia. Her father and brother have a distinct Russian accent. Various other minor human characters speak English with recognizable regional accents. Ambassadors Delenn and Londo Mollari, both alien characters, have distinct accents similar to Slavic. Delenn has actress Mira Furlan's normal Croatian accent; most other Minbari have native-speaker accents for English (for example, American for Lennier, British for Neroon). Londo's accent was developed independently by actor Peter Jurasik, and was imitated by William Forward, who played Lord Refa. Straczynski has described Londo's accent as being that of the "old school" of the Centauri Imperial Court.

Abbai The Abbai are a race from the fictional Babylon 5 universe. Their homeworld is Abbai 4 called SsumsshaA, which orbits the star Beta Pictoris in Sector 16. A peaceful, matriarchal society, the Abbai were the central force behind the formation of the League of Non-Aligned Worlds.Graw, B. and Glass, R.: Babylon 5 Wars Second Edition Rules Compendium, pages 211–216. Agents of Gaming, 2000 The Abbai have had interstellar flight since the Earth year 1461.

Brakiri The Brakiri feature heavily in the episode "Day of the Dead", which deals with a mysterious Brakiri religious celebration. The Brakiri have had interstellar flight since the Earth year 2111. Brakiri are primarily night dwellers, and have a reputation among the other races as being pack rats and scavengers. The Brakiri were among the first that Shadows attacked openly in Earth year 2260. They were also among those that were under attack by the Centauri in Earth year 2262.

Centauri Republic The Centauri Republic, once a mighty empire, has grown decadent and is in a slow decline. Their homeworld is Centauri Prime, a small Earth-like planet consisting of two large continents and several smaller islands divided by large oceans of water. The planet has a population of about 3.4 billion.

The Centauri, a humanoid race similar in appearance to humans, are a proud and aristocratic people governed by an emperor and a nobility-driven senate called the Centaurum. The Centauri were the first alien race to openly contact humanity. The Centauri initially tried to convince the humans that they were a long lost colony of the Republic. This ruse failed once human scientists got access to Centauri DNA, but Centauri-Human trade and contact led to tremendous technological advancement and humanity's eventual rise as a major power. For instance, it was the Centauri who sold jumpgate technology to the Earth Alliance, allowing humanity to start colonizing interstellar space.

Centauri males of high social status typically wear their hair in Peacock-tail shaped fans, the length and style of which are determined by relative social class. Low-class Centauri males have not been seen without helmets. Centauri females mostly or entirely shave their heads. In this episode Centauri emperor Turhan specifically states that Centauri females shave their heads, and his wives are shown to be completely bald.The Centauri Republic Factbook (op. cit., p. 8), which is non-canonical, incorrectly reports that only Centauri males grow hair at all. The social significance (if any) of complete versus almost-complete baldness on a female is not specified outright; however, in the televised episodes, older females are shown completely bald, and younger (up to the apparent human age range of the 30s, and often but not always unmarried) with ponytails. In this episode, the unmarried Centauri female Adira Tyree is shown to have a ponytail. In this episode, the youngest of Mollari's three Centauri wives has a ponytail, while the elder two are completely bald. In this episode, Vir Cotto's fiancée Lyndisty is shown to have a ponytail.

The cardio-vascular system of Centauri includes two hearts instead of the single heart that humans have. The right heart is a solid mass of muscle which provides most of the force behind the body's blood circulation. The left heart is much more complex—this heart is made up of an intricate system of thousands of veins that help cleanse the blood in a manner similar to the human kidneys. The sexual organs of the Centauri are in a different location than the human sex organs. Male Centauri have six tentacle-like genital organs that extend out from the sides of the body and "fold" in over the solar plexus when not in use. The males can stretch the tentacles out to four feet.

A popular Centauri meal is a dish called spoo, which mostly serves as a running gag in the show. Spoo comes from an animal of the same name which is "regarded as [one of] the ugliest animals in the known galaxy by just about every sentient species capable of starflight" according to character Emerson Briggs-Wallacewith.

The government of the Centauri Republic is, despite it being called a republic, ruled by an Emperor and an assembly of Ministers and heads of various Houses that form the Centaurum. The Centauri Republic is reflective of many imperial cultures on Earth, although J. Michael Straczynski makes a specific comparison: "the British Empire once upon a time... It was a great military power. But slowly, as can happen, they grew content, and lazy, and gradually their own empire began to slip between their fingers".

Dilgar The Dilgar were a race depicted in the show as an aggressive, warlike society who initiated a sudden and unexplained campaign of conquest against all neighboring worlds between 2229 and 2232. Regarding all alien species as little more than animals, the Dilgar slaughtered entire populations and ruthlessly enslaved the few survivors.

A coalition between The League of Non-Aligned Worlds and the Earth Alliance reversed the Dilgar's advances and ultimately blockaded them on their homeworld. This victory established Earth, previously considered a newcomer, as an interstellar power. The cause of the Dilgar's abrupt attacks was revealed when their sun went supernova, wiping out almost the entire species. Only a single civilian colony on the planet Planthos survived the war and cataclysm.

In the episode Deathwalker, a Dilgar survivor of the war appears on the Babylon 5 station in the year 2258 and is revealed to be Jha'Dur, known to many races as "Deathwalker" because of the many extravagant war crimes she committed. Following the war, Jha'Dur secretly entered the service of the "Wind Swords", a particularly militant Clan of the Minbari Warrior Caste. She came to Babylon 5 to establish her people's legacy via an immortality serum, claiming that it was so that all races could be indebted to the Dilgar for immortality. However, it is revealed that the immortality serum required the death of a sentient being to work: Deathwalker intended to increase war, bloodshed and other atrocities among the other races. She nearly accomplished her plan: an alliance of races attempted to employ her to refine a version of the serum that did not require murder; however, her Minbari flyer was destroyed by a Vorlon warship as she was leaving Babylon 5.

The Dilgar are depicted as humanoid with pointed ears similar to Vulcans. They are designed to have somewhat longer eyes than the eyes of species such as Humans, Centauri, Minbari, Narn etc. Their pupils are vertically elongated similar to those of cats. The bridge of their nose is widened and goes up the forehead, making a v-shape rise in the forehead. The tips of their eyebrows are pointed, similar to Centauri eyebrows.

Drakh Servants of the Shadows until the older race left Z'ha'dum, the Drakh became major antagonists late in the Babylon 5 series and in its sequel, Crusade. The Drakh were users of biotechnology and were responsible for releasing a nanovirus plague on Earth in A Call to Arms. The Drakh also grow parasitic extensions of their being, known as Keepers, on their bodies. The Keepers function as remote control systems for the Drakh's conscripted servants; they attach a Keeper to a host organism as a means of controlling and monitoring the person through telepathic relay.

Drazi The Drazi race in the show was depicted as reptilian in appearance, with blue skin, thick skull bones, and no nostrils, although they are otherwise humanoid. They were a member of the League of Non-Aligned Worlds and are members of the Interstellar Alliance.

Drazi have a pouch in one of their armpits; it is actually used as a reproductive area. In Crusade, the Drazi are described as hermaphrodites.

The ancient Drazi used to live outdoors, only sleeping inside at night. As a result, Drazi houses are constructed with large balconies and a much smaller interior. The streets on the Drazi homeworld are very narrow, reflecting their long history of warfare: armoured Drazi vehicles found it impossible to invade an enemy city without first demolishing its buildings.

Every five Earth years, the Drazi have a "civil war" between two rival factions, green Drazi and purple Drazi. A Drazi's allegiance is decided arbitrarily by choosing a colored sash, either green or purple, which is then worn around their torso. There is no inherent motive or reason for the civil war at all, other than tradition. This "civil war" takes the form of a series of (usually) non-lethal contests. A group of Green Drazi on their homeworld intended to escalate to lethal violence, and was quickly followed by Green Drazi on Babylon 5. Commander Susan Ivanova ended the contest through the use of a loophole in the rules: taking the sashes of the leaders and ordering both groups to stop fighting. In the fifth season episode The Ragged Edge, a Drazi is shown wearing a sash made up of both colors on the Drazi homeworld.

The Drazi participated heavily during the Shadow war in 2260, and its later escalation with the full of entrance of the Vorlons by early 2261.

The only known major warship seen on-screen operated by the Drazi is the "Sunhawk", a small, highly maneuverable cruiser possibly designed primarily for patrols and quick hit-and-run attacks on larger targets. A Sunhawk has also been shown to have the ability to carry and launch a small one-man craft; during the battle of Coriana 6 one is seen flying vertically while firing a pulse weapon. This small vessel is designated the "Sky Serpent." In the B5 wars board game the Sky Serpent is classed as a superheavy fighter.