| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Roosevelt_Academy | A | NT | |
| 3 | Frostproof |
A | 46.49 | Polk County Championship 2025 |
| 4 | Hardee |
A | 47.37 |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Okeechobee |
A | 45.29 | Sebring Relay Meet 2025 |
| 2 | LaBelle |
A | DNS | FHSAA 3A District 11 |
| 3 | Sebring |
A | 42.15 | FHSAA 3A Region 2 |
| 4 | Avon Park |
A | 43.78 | FHSAA 2A District 9 |
| 5 | DeSoto County |
A | 44.07 | Avon Park Invitational |
| 6 | Lake Placid |
A | 46.40 | Sebring Relay Meet 2025 |