| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Lake Placid |
A | 4:04.98 | Lake Placid Meet #2 |
| 3 | DeSoto County |
A | 3:55.02 | Sebring Relays 2024 |
| 4 | Sebring |
A | 3:37.94 | Clewiston Big Lake Challenge 2024 |
| 5 | LaBelle |
A | DNS | Edison Relays 2024 (Approved Sanctioned Meet) |
| 6 | Avon Park |
A | 4:02.72 | Lake Placid Meet #1 |
| 7 | Okeechobee |
A | 4:16.20 | Lake Placid Meet #2 |