| Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note | |
|---|---|---|---|---|---|---|---|---|
| 1 | South Lake |
A | Madden | Parker | Thompson | Cypre | 8:41.17 | FHSAA 4A District 4 |
| 2 | Wiregrass Ranch |
A | Stebleton | Willis | Ammerlaan | Gregory | 9:00.00 | |
| 3 | Fivay |
A | Oliver | Brown | Luna | Rodriguez | 9:26.21 | West SAC Conference Meet |
| 4 | Anclote |
A | Cruz-Torres | Miranda | Lightfoot | Sanchez | 9:53.04 | Anclote Sharks T&F Invitational 2024 |
| 5 | Angeline Academy | A | Lingala | Gawienczuk | Arun | Griffith | 10:25.28 | 2025 Newsome Wolves Invitational |