Every Spider-Man villain ever or just the Sinister Six?
Alistair Smythe
Beetle
Big Man (Frederick Foswell)
Big Wheel
Black Cat
Boomerang
Calypso
Carnage
Carrion
Chameleon
Clash
Circus of Crime
Crime-Master
Doctor Doom
Doctor Octopus (Otto Octavius)
Doctor Octopus (Carolyn Trainer) aka Lady Octopus
Doppelganger
Electro
Enforcers
Finisher
Gibbon
Goblin Nation
Green Goblin (Norman Osborn)
Green Goblin (Harry Osborn)
Grizzly
Hammerhead
Hobgoblin
Human Fly
Hypno-Hustler
Hydro-Man
Jackal
Jack O'Lantern and Mad Jack
Juggernaut
Kangaroo
Killer Shrike
Kingpin
Kraven the Hunter
Lightmaster
Living Brain
Lizard
Looter
Man-Wolf
Mendel Stromm
Mister Negative
Molten Man
Morbius the Living Vampire
Morlun
Mysterio
Prowler
Puma
The Queen
Rhino
Ringer
Rose (Richard Fisk)
Rocket Racer
Sandman
Scorcher
Scorpion
Shocker
Shriek
Silvermane
Silver Sable
Sin-Eater
Sinister Six
Sinister Syndicate
Spencer Smythe
Speed Demon
Spider-Slayers
Spot
Squid
Stegron
Stilt Man
Swarm
Tarantula
The Burglar who killed Uncle Ben
Tinkerer
Titania
Tombstone
Trapster
Venom
Vermin
Vulture
White Rabbit
Will o' the Wisp
Wraith
The only two I can see presenting a serious challenge to Bats are Doom and the Juggernaut. That being said, even with a week in between battles, Batman is going to start taking damage at some point that he can't fully recover from in a week. I can't see him working his way too far down this list.