| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Bloomingdale |
A | Golden | Donovan | Owen | Williams | 10:31.84 | Newsome Wolves Senior Night 2025 |
| 2 | Riverview |
A | Nix | Fernandez | Todd | Vesga | 10:31.87 | FHSAA 4A District 11 |
| 3 | Lennard |
A | Vargas | Urbina | Smith | Torres | 10:38.47 | 2025 Vernon Korhn Hillsborough County Championships |
| 4 | Morgan High School | A | James | Jones | Hancock | San Martin | NT |