Bimbo's Initiation has an underground cult send the title character through a series of dangerous traps, many of which are strange enough to be a little creepy. Of course, the creepiest aspect of the whole picture is the cult itself, as they are literally called the Do-It-Or-Die.
Minnie the Moocher stands out among a whole collection of creepy and surreal Fleischer cartoons. The main number is sung by an eerily rotoscoped ghost walrus, and we are shown scenes of black-eyed ghost cats drinking their mothers milk, eventually sucking her completely dry and possibly killing her, skeletons drinking beer and collapsing, and ghosts getting the electric chair. If all that wasnt enough to give you the chills, one gigantic witch flies in and provides us with one heck of a Jump Scare.
The titular villain of The Old Man of the Mountain is implied to be a rapist, since Betty ends up encountering a crying woman who reveals that her babies look identical to the Old Man. The fact that he later pursues Betty with obviously malicious intentions does not help his case at all.
Red Hot Mamma is about Betty going to Hell and meeting Satan.
The evil spider in Cobweb Hotel is one of the more threatening villains from these shorts. Probably the most notable moment of his is during his Villain Song when he shows off his screaming guests after leering menacingly at the camera.
Satisfying as it may be, Greedy Humpty Dumptys demise in the short of the same name is quite intense, especially for young children.
Play Safe takes its message very seriously, and while the kids bizarre dream of the trains coming to life is pretty darn creepy, the scenes where he is in danger of being crushed by the actual trains are scary for a different reason.
Small Fry has an effective Disney Acid Sequence that is a lot of fun, but also quite unsettling thanks to the dark setting and the very large and very carnivorous denizens of the deep menacing our protagonist.