Adam Seymour "Queer Duck" Duckstein
- Voiced by: Jim J. Bullock
"I'm gay as a goose!"
The show's main character, an out and proud duck who thoroughly enjoys flirting with and sleeping with every man he can.
- All Gays are Promiscuous: He's not really content with having sex with just one man. "Wedding Bell Blues" even has him marry Openly Gator while giving a long list of circumstances where he might cheat on Openly Gator with another man.
- Cure Your Gays: He is made straight by Reverend Vandergelding in the movie just before his wedding to Lola Buzzard. This doesn't last, as shortly after Lola's demise, her manservant Peccary and Barbara Streisand help make Queer Duck become gay again.
- Camp Gay: He's openly gay, has a lot of feminine mannerisms and wears a rainbow shirt.
- The Gadfly: He lives for making a fool out of homophobes.
- Half-Dressed Cartoon Animal: Much like a lot of cartoon ducks, he doesn't wear pants.
- Happily Married: He marries his boyfriend Openly Gator by the end of the episode "Wedding Bell Blues" and Queer Duck: The Movie.
- Interspecies Romance: He's a duck whose main significant other is an alligator and in fact rarely shows attraction to any men who are also waterfowls.
- Transparent Closet: The very first episode has him decide to come out at work. This is met with awkward silence and one of the people in the waiting room remarking "Well, duh". His nephew Little Lucky makes a similar remark in "A Gay Outing" after his friends point out his uncle's sexuality.
Steven Arlo "Openly" Gator
- Voiced by: Kevin Michael Richardson
Queer Duck's boyfriend and later husband.
- Accidental Misnaming: Queer Duck's father misnames him as Openly Crocodile in "Homo for the Holidays".
- Half-Dressed Cartoon Animal: He doesn't wear pants.
- Happily Married: He and Queer Duck get married in "Wedding Bell Blues" and the movie.
- Interspecies Romance: He's an alligator whose life partner is a duck.
- Non-Mammalian Hair: He has a jewfro in spite of being a reptile.
- Species Surname: A gator with the surname Gator.
Bi-Polar Bear
- Voiced by: Billy West
A polar bear with a habit of telling bad jokes.
- Anal Probing: When abducted by aliens in "Tales of the City Morgue", he begs the aliens to probe him, to which they reluctantly oblige.
- Half-Dressed Cartoon Animal: He wears a hat and a vest, but no pants.
- Laughing at Your Own Jokes: He's the only one who ever laughs at his jokes.
- No Celebrities Were Harmed: His voice is an impression of Paul Lynde.
- Non-Indicative Name: The "Bi-Polar" in his name is supposedly a play on "bisexual", but he's clearly only attracted to men and is even outright offended at an alien taking on the form of a curvaceous woman in "Tales of the City Morgue".
Oscar Wildcat
- Voiced by: Maurice LaMarche
A cultured gay cat.
- Deadpan Snarker: He is very clearly the most sardonic of the main characters.
- Everyone Has Standards: In "Tales of the City Morgue", he is horrified by his Frankenstein's Monster being a flagrant gay stereotype.Oscar Wildcat's monster: (singing) Hello, Frisco, hello!
Oscar Wildcat: Dear God! Is it possible to be too gay?! - Half-Dressed Cartoon Animal: He doesn't wear pants.
- Interspecies Romance: The movie has him hook up with a male dog named Rex, who he initially knew as a woman named Regina due to Rex being an exceptionally skilled drag artist.
- Matricide: A minor recurring gag has him scheming to kill his own mother.
- No Celebrities Were Harmed: His name is an obvious play on 19th century author Oscar Wilde.
Dr. and Estelle Duckstein
- Voiced by: Maurice LaMarche (Dr. Duckstein), Estelle Harris (Estelle Duckstein)
Queer Duck's parents, who aren't that accepting of their son's sexuality.
- Abusive Parents: They're not physically abusive, but they do subject him to a lot of verbal abuse, plus their refusal to accept his homosexuality doesn't help.
- I Want Grandkids: Dr. Duckstein states in the first episode that he wants Queer Duck to give him grandchildren, disregarding that his son isn't attracted to women. It's not known what he thinks of his grandson Little Lucky from his other son Lucky.
- Racist Grandma: "Wedding Bell Blues" has Estelle make it clear that she's not happy with her son not just marrying another man, but a man who is a reptile and Gentile. This is played more for laughs in the movie, where they're both shocked to learn that Michael Jackson is a black man and Estelle later tells Jackson not to cry at Lola Buzzard's funeral because he'll become black again.
Lucky Duckstein
- Voiced by: Ru Paul
Queer Duck's straight older brother.
- Big Brother Bully: It's implied he frequently picked on Queer Duck when they were younger.
- Captain Oblivious: He is apparently unaware that his younger brother is gay.
- Chuck Cunningham Syndrome: He isn't acknowledged anymore after his debut episode, to the point that "Quack Doctor" has a flashback to the day Queer Duck was hatched that indicates that Queer Duck is the Duckstein's only son, plus a remark Openly Gator makes upon meeting Queer Duck's sister Melissa in "Homo for the Holidays" suggests that Melissa is Queer Duck's only sibling. He doesn't even appear in the movie's flashbacks to Queer Duck's youth.
Little Lucky
- Voiced by: Tress MacNeille
Queer Duck's nephew and Lucky's son.
- Chuck Cunningham Syndrome: He only appears twice and is never brought up again. Oddly enough, his second appearance in "A Gay Outing" premiered not long after "Quack Doctor", the episode that contradicted the existence of his father.
- Missing Mom: We never see his mother.
- Innocent Inaccurate: His debut episode has him mistake his uncle's porn tapes of Poke-A-Man and Rock Hard and Bull Sprinkle for tapes of Pokémon: The Series and Rocky and Bullwinkle.
- Ping Pong Naïveté: In spite of the aforementioned mistaking videotapes of his uncle's gay porn for copies of the kid-friendly media they're spoofing, he responds to his friends pointing out that his uncle is gay in "A Gay Outing" by indicating that he already knew about his uncle's sexual orientation.
Melissa Duckstein
- Voiced by: Tress MacNeille
Queer Duck's lesbian sister.
- Butch Lesbian: She is attracted to women and has a masculine appearance, to the point that Openly Gator initially mistakes her for being Queer Duck's brother.
- Interspecies Romance: Her girlfriend Yvette is literally a beaver.
- Visual Pun: Her girlfriend's a beaver, which is an overt pun on the fact that "beaver" can be slang for the female genitalia.
Dr. Laura Schlessinger
- Voiced by: Tress MacNeille
A fictional representation of real-life homophobe Dr. Laura Schlessinger, who is the web series' most frequent antagonist.
- Demoted to Extra: Her only appearance in the movie is a cameo where her photograph is one of the ingredients used in Reverend Vandergelding's potion he creates to cure Queer Duck of being gay.
- Eye Scream: In "Queer Doc", she makes the mistake of peeking through a glory hole, and you can probably guess what poked her in the eye.
- Heteronormative Crusader: She's a fictional representation of a real-life homophobe, so of course she's an activist against gay rights.
- Karmic Butt-Monkey: She suffers a lot of humiliating injuries, but given that she is a raging homophobe, she really has it coming.
- Off with His Head!: In "Mardi Foie Gras", Queer Duck arranges for a voodoo curse that causes her head to fall off mid-sentence.
- Politically Incorrect Villain: A given, since she's constantly attacking Queer Duck for being gay.
- Take That!: Not surprisingly, the web series shows her in a decidedly negative light, with Queer Duck accusing her doctorate of being phony and subjecting her to all sorts of over-the-top punishment in retribution for her homophobia.
Fairyland Mascot
- Voiced by: Kevin Michael Richardson
The mascot of the gay-friendly theme park Fairyland. Appears in the movie as the homophobic mascot of Happyland.
- Adaptational Jerkass: While his debut in the web series episode "The Gayest Place on Earth" already depicted him as rather mean due to yelling at Queer Duck for hitting on his lover and later admonishing said lover for not finishing a blowjob, the movie reinterprets him as an out-and-out homophobe.
- Adaptational Sexuality: He is in a relationship with a male turkey named Gobble the Salty Seaman in the web series episode "The Gayest Place on Earth", but in the movie is supposedly straight, as he isn't shown to be interested in men and acts homophobic towards Queer Duck and the other gay men he's brought to Happyland, plus the end of the film has Queer Duck respond to his earlier "Beat it, you homos" statement by saying "Beat it, hetero".
- Crazy Jealous Guy: He doesn't take kindly to Queer Duck flirting with his "first mate" Gobble the Salty Seaman.
- Laser-Guided Karma: His homophobia in the movie is punished when Queer Duck buys out Happyland, turns it into Fairyland and has the homophobic mascot kicked out.
- Mocky Mouse: He is blatantly a pastiche of Mickey Mouse, right down to the round, black ears and high-pitched voice.
- No Name Given: He isn't given a specific name.
- Politically Incorrect Villain: The movie depicts him as a homophobe who refuses admittance to gay men and yells "Beat it, you homos!"
Lola Buzzard
- Voiced by: Jackie Hoffman
An aged former star who befriends Queer Duck in the movie.
- The Beard: Her bonding with Queer Duck leads to her wanting to marry him. When Queer Duck points out that he doesn't think the marriage will work because he's gay, Lola replies that she doesn't mind because a lot of her previous husbands were gay.
- False Teeth Tomfoolery: She is revealed to be wearing dentures when she and Queer Duck attempt a Spaghetti Kiss.
- Interspecies Romance: She is a buzzard and is mentioned to have been married to Peccary, a pig. This isn't including how she marries Queer Duck and has sex with him due to him being converted to straight by Reverend Vandergelding.
- Non-Mammal Mammaries: Being a buzzard doesn't prevent her from having huge bazooms.
- Out with a Bang: She falls over dead the morning after having sex with the made-straight Queer Duck.
- Pretentious Pronunciation: The second part of her surname is pronounced like "hard".
- Serial Spouse: She is stated to have married a lot of times.
- Species Surname: A buzzard with the last name Buzzard.
Peccary
- Voiced by: Tim Curry
Lola Buzzard's manservant and chauffeur as well as her second and fifth husband. He is an anthropomorphic pig.
- Ambiguously Bi: He is said to be an ex-husband of Lola Buzzard and states that he loved her after she dies, but a later scene has him flirt with a male hot dog vendor.
- I Need a Freaking Drink: When annoyed by Lola changing her mind on whether or not she wants any wine, he takes a big swig from the bottle after going back to Lola's end of the table.
- Interspecies Romance: He's a pig and was Lola Buzzard's second and fifth husband.
Reverend Vandergelding
- Voiced by: Jeff Bennett
An anthropomorphic horse and religious fundamentalist who attempts to make Queer Duck straight. He is the main antagonist of the movie.
- Big Bad: He is responsible for a lot of the conflict in the movie, as his efforts to make Queer Duck straight strains Queer Duck's relationship with Openly Gator and cause Queer Duck to have issues accepting his sexuality.
- Evil Plan: His ultimate goal is to make Queer Duck straight. When that gets undone, he switches to using his straight serum to make all the gay men at the Fairyland theme park straight.
- Heteronormative Crusader: Like any other religious fundamentalist, he views homosexuality as an illness and wants to make every gay person straight.
- Karmic Rape: His final fate is getting advances from a bull at a gay rodeo, which is treated as his comeuppance for attempting to make gay men straight.
- No Song for the Wicked: The movie is an Animated Musical and he is one of the only characters in it who doesn't get to sing (aside from some parts of Queer Duck's song about how he came out to his parents).
- Politically Incorrect Villain: Like just about every Queer Duck villain, he's a homophobe.
- Reused Character Design: He is identical to the unnamed homophobic preacher from the web series episode "Oh Christ!", the leather-clad man in the gay bar's restroom in "Queer Doc" and the flight attendant from the episode "Wedding Bell Blues".
