Here is the lineup used for the 74th Annual Macy's Thanksgiving Day Parade, which took place in 2000.
- Mickey Mouse (The Walt Disney Company) (Retired) - 1st and only time
- Blue (Nickelodeon) - 2nd time
- Big Bird (Sesame Workshop) (Retired) - 13th and last time
- Rugrats (Nickelodeon) - 4th time
- Jeeves (Ask.com) - 1st time
- Honey Nut Cheerios Bee (General Mills) - 2nd time
- Ronald McDonald (McDonald's Corporation) - 1st time
- Millennium Snoopy (United Media) - 2nd time
- Arthur (Random House Children's Media Group) - 4th time
- Rocky and Bullwinkle (Universal Studios) (Retired) - 3rd and last time
- Dexter (Cartoon Network) (Retired) - 3rd and last time
- Dragon Tales' Cassie (Sony Pictures Family Entertainment) - 1st time
- Barney the Dinosaur (Lyrick Studios) - 7th time
- Clifford the Big Red Dog (Scholastic) (First time since 1996) - 7th time
- Strawberry/Pistachio Ice Cream - 14th time
- White Macy's Stars (Red text) - 15th time
- Flying Fish - 4th time
- White Macy's Stars (Green text) - 15th time
- Americana Spheres - 2nd time
- Basketball and Football - 10th time
- Harold the Fireman - 5th time
- Cloe the Holiday Clown - 7th time
- Snowstars (Retired) - 13th time
- Tom Turkey
- Blue's Clues
- Friends for Life
- The Three Little Pigs
- The Old Lahaina Luau
- Flights of Fancy (Retired)
- Mother Earth
- Amazing Creatures of the Wild
- Soaring Spirits Canoe
- The World of Wiggle (Retired)
- M&M's Network
- Fifty State Quarter Stagecoach
- Wakin’ Up (Retired)
- Statue of Liberty
- The Ship of State
- Pep Rally
- Road to the Future
- Santaland Express
- Simple Simon's Fair
- Barney's Night Before Christmas
- Winter Holiday (Retired)
- Rudolph the Red-Nosed Reindeer
- Santa's Sleigh
- Dog Catcher
- Rolling Bath Tub
- Jamaica Folklore Rod Puppets
- Big Red Shoe Car
- New York City Police Department Mounted Patrol
- The Corrs; performed "Breathless"
- John Ritter
- Emeril Lagasse
- Steve Burns
- The Cast and Muppets of Sesame Street
- Ann Hampton Callaway; performed "One Brick at a Time"
- Old Lahaina Luau Hula Dancers
- Miss America 2001 Angela Perez Baraquio
- Joe Torre
- The New York Yankees
- Sara Evans
- Jennifer Day
- Kenny Chesney; performed "Me and You"
- Jeff Corwin
- Baha Men; performed "Who Let the Dogs Out"
- Jo Dee Messina; performed "Dare to Dream"
- Ronald McDonald and The McKids
- Innosense; performed "Say no More"
- Tito Puente Jr.; performed "Dance Dance"
- LFO; performed "West Side Story"
- Nikki McCray
- The Cast of Passions
- BBMak; performed "Still on your Side"
- Rascal Flatts; performed "Praying for Daylight"
- Aaron Carter; performed "I Want Candy"
- Barney, Baby Bop and B.J.
- Mikaila; performed "So in Love with Two"
- Andrea McArdle; performed "Rudolph the Red-Nosed Reindeer"
- Santa Claus and Mrs. Claus
- Lincoln High School Patriot Marching Band
- Ohio University Marching 110
- Blue Springs Golden Regiment Marching Band
- Walterboro High School Band of Blue
- Spirit of Waxahachie Indian Band
- "Mighty Sound of Maryland" Marching Band
- Clovis West High School Marching Band
- Elko High School Pride of Nevada Marching Band
- Homewood High School Patriot Band
- Science Hill High School Topper Band
- NYPD Marching Band
- Cheshire High School Marching Ram Band
- Bergenfield High School Marching Band
- Western Clowns
- Bowling Hall Clowns
- Circus Clowns
- Funny Firefighter Brigade
- America Sings!
- Mike Miller Dance Team
- Dallas Tap Dazzlers
- Starmakers Dance Company
- Texas University Strutters
- Universal Cheerleaders Association
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