| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | East Ridge |
A | Walker | Moore | Mccleary | Lederer | 8:35.00 | |
| 2 | Dr. Phillips |
A | Jean Baptiste | Medley | Caceres | Mescharles | 8:38.24 | 8:38.24 Metro West Championships |
| 3 | Kathleen |
A | Chappell | Morales Torres | Smith Jr. | Williams | DNS | Kowboy Invitational |
| 4 | Frostproof |
A | Ramirez | Barajas | Valencia Calixtro | Rocendo | 9:13.13 | FHSAA 2A Region 3 |
| 5 | Davenport |
A | Diaz DeJesus | Locker | Valladares | Thomas | 9:41.59 | 9:41.59 Polk County Championship 2023 |
| 6 | Mulberry |
A | Cortez | Dilworth | Martinez | Jones | DNS | 9:43.81 Polk County Championship 2019 |
| 7 | Auburndale |
A | Bowman | Davis | Jordan | Mintz | NT |