| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Okeechobee |
A | 4:04.98 | Falcon Classic 2024 |
| 2 | DeSoto County |
A | 3:46.79 | Avon Park Meet #1 2025 |
| 3 | Sebring |
A | 3:33.28 | FHSAA 3A District 8 |
| 4 | Sebring |
B | 3:33.28 | FHSAA 3A District 8 |
| 5 | Avon Park |
A | 3:56.62 | Sebring Invitational 2024 |
| 6 | Okeechobee |
B | 4:04.98 | Falcon Classic 2024 |