| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Okeechobee |
B | Bloom | Perdomo | Maitland | McQueen | 54.33 | Jensen Beach Invitational 2025 |
| 3 | LaBelle |
A | Sandoval | Sandoval | Jackson | Williams | 52.32 | Sebring Meet #2 2025 |
| 4 | Okeechobee |
A | Barrett | Aponte | Nesby | Rochelle | 54.33 | Jensen Beach Invitational 2025 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Clewiston |
B | Bradford | Rodriguez | Benjamin | Bredwood | DNS | Sebring Meet #2 2025 |
| 4 | Clewiston |
A | McCain | Stephens | Miscaide | Bradford | DNS | Sebring Meet #2 2025 |
| 5 | Moore Haven |
A | Herrera | Myers | Hughes | Faison | DNS | Sebring Meet #2 2025 |