| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | South Fork |
A | DNS | Bulldog Invitational 2025 |
| 3 | Fort Pierce Central |
A | 3:36.21 | Cobra Relays 2025 |
| 4 | Jupiter |
A | 3:35.10 | Bulldog Invitational 2025 |
| 5 | Port St. Lucie |
A | 3:35.73 | SLC Championships 2025 |
| 6 | Martin County |
A | 3:41.09 | Bulldog Invitational 2025 |
| 7 | Fort Pierce Westwood |
A | DNS | |
| 8 | Lincoln Park |
A | NT |