| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Sarasota Military Academy |
A | 46.48 | Lakewood Ranch Invitational 2025 |
| 4 | Cardinal Mooney |
A | 42.12 | Guy Thomas Memorial -Sponsored by Anderson Electric of SWFL |
| 5 | Winthrop College Prep | A | 44.88 | Raven Invitational 2025 (Sanctioned) |
| 6 | Out-of-Door |
A | DQ | North Port Invitational 2025 |