"[T]he dynamic duo wont bother with us. We're already in prison All theyll be doin is rounding up other criminals. More talent for the Gotham state pen gang .The harder they work, the more they help us, so I say long live Batman and Robin."
— The title character, "Ma Parker"
By order of appearance.
- The Riddler is a stylish thief and kidnapper who has few scruples about his targets. He loves leaving clues about his crimes for Batman but excels at misdirection and isn't above using those clues to lure the heroes into traps or tricking Batman into making a false arrest to sue him in court. He nearly finds the Batcave by having an accomplice impersonate Robin. When an eccentric tycoon hires Riddler to make a silent film of himself fighting Batman, he takes to the task with great enthusiasm, while also planning to rob his employer. Even when he's at his worst and plants bombs to extort money from the city, he is willing to disarm them once he is paid. Elegant, witty, and devilishly clever, this version of the Riddler is responsible for making the character a Breakout Villain.
- Catwoman is a conniving Classy Cat-Burglar, extortionist and, on occasion, would-be world conqueror. Her trademark cat o' nine tails whip and clawed gloves act as both costume accessories and useful tools in her robberies. She engages in multiple alliances with fellow super villains and is always an intelligent and assertive figure in those partnerships, such as when she arranges for some Jury and Witness Tampering after she and the Joker are captured. She prefers using her talents for planning and misdirection but will also fight back against the Caped Crusaders when she's cornered. Catwoman has a strong mutual attraction to Batman and is willing to spare Batman's life even when she gets the drop on him and gracefully accepts her loses to him. On multiple occasions, Catwoman manages to escape from Batman by faking her death. Whenever she is arrested, she puts on a convincing show of being reformed that fools even people who should know better, although the possibility she will genuinely reform one day is a very real one.
- "Zelda the Great" & "A Death Worse than Fate":
- Zelda the Great is a renowned escape artist who robs a bank every year on April Fools' Day to finance her high-profile Magic act. When Batman and Robin trick her into thinking that the proceeds of her latest robbery are worthless counterfeit bills, she sets out to gain her illicit funds through other means. She easily sees through and foils a trap they set for her and kidnaps Aunt Harriet for ransom, but lets her go unharmed as soon as Batman admits that the money from the bank was real. Zelda leaves a clue in Harriet's pocket for Batman to find so that she and her gadget maker can put him in the latest escape device and force him to test it. When Zelda learns that her partner plans to kill Batman and Robin, she saves their lives, gracefully turns herself in, and reforms while in prison.
- Eivol Ekdal is a shady inventor who makes the death traps Zelda becomes famous escaping from. He helps plan the robberies she uses to pay him. To test a complex trap, he seeks to exploit the Caped Crusaders' reputation for escaping from them. He collects $100,000 from two hitmen who want to kill Batman and has them hide nearby so that they can shoot Batman and Robin as soon as they escape his trap. Ekdal comes dangerously close to killing the heroes through fairly mundane means, and only fails due to Zelda's change of conscience.
- "True or False Face" & "Holy Rat Race":
- False Face is a master criminal known for his ability to assume convincing disguises in an instant. During a counterfeiting caper, he kidnaps and impersonates Chief O'Hara, regularly rescuing his thugs within seconds every time the police arrest them. False Face sets an unusually effective death trap for the Dynamic Duo that only fails due to the betrayal of his assistant, Blaze. When the heroes thwart his final robbery attempt, False Face repeatedly refuses to admit defeat and flees from them by motorcycle, then uses a rapid series of disguises and misdirection when that fails. Once his array of tricks runs out, False Face compliments Batman on his victory while threatening to return in the future.
- Blaze is False Face's aide, who is just as well-versed in disguise and sudden, audacious escapes as her employer. She has a talent for carrying around seemingly innocent items with hidden gadgets that she uses to further False Face's plans, such as using knockout gas in her hair clip against Robin. She lures Batman and Robin into a trap, but balks at False Face's plan to kill them and quickly contrives a way to save the Caped Crusaders' lives, even remaining defiant when taken captive by her former boss. After Batman and Robin rescue her, she helps thwart False Faces escape. The two-parter ends by showing a reformed Blaze meeting with her parole officer Bruce Wayne and favorably comparing him to Batman.
- "The Bookworm Turns" & "While Gotham City Burns": Lydia Limpet is a devoted lover and accomplice of the literary-themed super villain, the Bookworm. She assists Bookworm in a plan to fake Commissioner Gordon's assassination to lure the heroes to police headquarters and booby trap their car. When this fails, she poses as a hostage to send them on a wild goose chase. When they figure out what she's up to, she realizes that they know she's a Decoy Damsel when Robin calls her by name when she doesn't remember introducing herself. She tricks him into opening a booby-trapped book and warns the Bookworm to abandon his hideout and put Robin in a Death Trap. Later, she helps trap Batman and Robin inside a giant, magnetized oven that resembles a cookbook.
- "The Greatest Mother of Them All" & "Ma Parker": Ma Parker and her four adult children are an infamous outlaw family who have committed bold heists throughout the country without being caught. When they come to Gotham City, they have accomplices inflate the local prison as guards and inmates. They launch a crime wave designed to attract Batman's attention and cause him to arrest them, while acting like they're on the run and gamely resisting arrest. Upon being dropped off at prison, they have a trustee put a bomb in the Batmobile and take the warden prisoner while setting themselves up in a Luxury Prison Suite. They plan to use the prison as a hideout while committing robberies. When Batman and Robin survive the attempt on their lives, Ma decides to leave him alone so the more people he arrests, the more recruits their gang gets. When they capture Batman and Robin, they make sure to keep an eye on them so they have no chance to escape the trap and take their eventual defeat with remarkable grace, remaining a loving family even in jail.
- "The Devil's Fingers" & "The Dead Ringers":
- Chandell is a renowned pianist who is driven to pull a years-long string of robberies after his brother Harry blackmails him over an incident where Chandell used a self-play piano during a White House concert after injuring his hand. He arranges for robberies to occur during his concerts to make himself look like a victim and make the authorities divert massive numbers of officers as security guards while leaving other locations unguarded. Chandell plans to woo and marry Aunt Harriet to get his hands on money from the Wayne fortune to pay off Harry and end his existence as as blackmailed criminal. He fakes an attack on himself and then tells Batman about Harry and his hideout so that they'll either fall into his brother's trap and stop threatening his plans, or capture Harry and remove the threat he poses. Chandell is poised to win Harriet's heart when he is betrayed by Harry and their confederates, ending the episode after his arrest gracefully playing the piano for his fellow inmates in prison.
- Doe, Rae, and Mimi are Chandell's lovers and accomplices. They carry out the robberies that help Chandell pay his blackmailing brother out of love. They use bagpipes with sleeping gas to rob Chandell, Harriet, and Alfred and throw suspicion off Chandell. When Batman and Robin raid Harry's hideout, the three women throw themselves into the middle of the fight to take advantage of Batman and Robin's Wouldn't Hit a Girl tendencies, helping buy time for their associates to turn the tide of the battle and place the heroes on a Conveyor Belt of Doom. Upon hearing of Chandell's plans to marry Harriet for her money, the trio betray and incapacitate Chandell, suspecting that he plans to toss them aside. Doe then helps Harry kidnap and nearly kill Harriet, and the three attempt to use their bagpipes against the heroes during the climax before being gassed themselves, last seen listening to a jailed Chandell's music, implied to regret betraying him.
- "The Puzzles are Coming" & "The Duo is Slumming": The Puzzler is an aviation-obsessed criminal who loves quoting William Shakespeare and dares the heroes to catch him with clues derived from the bard's plays. He convinces a wealthy tycoon that he has a legitimate business proposition to rob the man, meaning to steal an experimental military plane and then ransom it back to its builder even though a foreign power would pay more, because "I may be an arch-villain, but I am a one hundred percent naturalized American arch-villain!" After being captured, he takes complete responsibility for his plan's failure and respectfully salutes Batman for the depth of his Shakespeare knowledge.
- "Ring around the Riddler" & "The Siren's Wail": Lorelei "The Siren" Circe is the show's only meta-human villain with a singing voice that can hypnotize men. After assisting Riddler in a gambit to hypnotize several boxers, she embarks on her own mission to turn Gotham into a city of chaos. The Siren attempts to expose Batman's identity as a pre-emptive strike, hypnotizing Commissioner Gordon to be her catspaw. She also hypnotizes Bruce Wayne into signing over his entire fortune to her as a war chest for her campaign, then orders him to commit suicide. The Siren is left dangling from a ledge after a fight and Robin refuses to save her unless she sings a higher note that will cure Bruce's hypnosis but leave her mute, keeping her promise even after being rescued.
- "The Great Escape" & "The Great Train Robbery": Calamity Jan is introduced using a tank to break her boyfriend Shame out of a prison that he thought couldnt be breached. Jan and her mother, Frontier Fanny, and Shame assembled a gang to rob millions of dollars in old bills from a mail train. She never directly engages in physical combat with the heroes but is no Neutral Female, throwing things at them, being ready to use her gun, and seemingly fleeing a fight only to return with a gas that incapacitates the heroes. Calamity Jan also urges her boyfriend to shoot Batgirl rather than use her as a hostage and tries to convince him not to fall into an obvious trap. Jan also shows a softer side by dreaming of using the train loot to retire from crime and live on a ranch with her mother and Shame, and insists on rescuing Fanny from custody. Jan stands out as one of the few lovers of a villain who is his equal rather than just an accessory.
- "Minerva, Millionaires, and Mayhem": Minerva is rare among villains in that she doesn't make her criminal nature known to the public. She picks the brains of her clients to find out how to steal their valuables but remains trusted and respected by Commissioner Gordon and most of the city's millionaires. When Batman and Robin investigate her, she has them trapped in one of her machines and tries to boil them alive. After they survive, she convinces Gordon that it was a freak accident and prepares for one last job before fleeing the city. When that job turns out to be a trap, Minerva escapes from Batman but not the arriving police. An unfazed Minerva declares that she will make a magnificent new spa at the prison and lets the police take her away.