| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | South Fork |
A | DNS | Dr Garcia Invite #1 2025 |
| 3 | Martin County |
A | DNF | Jensen Beach Invite 2025 |
| 4 | Fort Pierce Westwood |
A | 49.45 | Bill Wilson Invitational 2025 |
| 5 | Lincoln Park |
A | 50.95 | Bill Wilson Invitational 2025 |
| 6 | Fort Pierce Central |
A | 51.80 | Cobra Relays 2025 |
| 7 | St. Edward's |
A | 55.43 | Viera High School Invitational |