| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Avon Park |
A | 4:02.72 | Lake Placid Meet #1 |
| 2 | Lake Placid |
A | DNS | HARDEE HS INVITATIONAL |
| 3 | Sebring |
A | 3:37.94 | Clewiston Big Lake Challenge 2024 |
| 4 | Hardee |
A | 3:42.46 | HARDEE HS INVITATIONAL |
| 5 | DeSoto County |
A | 3:55.02 | Sebring Relays 2024 |
| 6 | Moore Haven |
A | NT |