| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Centennial |
A | Dimarco | Kirkwood | Bryant Jr. | Mathew | 8:28.10 | Cobra Relays 2025 |
| 2 | Martin County |
A | Orellana | Slate | Lopes | Saracina | 8:36.80 | Cobra Relays 2025 |
| 3 | Fort Pierce Central |
A | Randolph | O'Dwyer | Pascoe | Rivera | 8:58.90 | Cobra Relays 2025 |
| 4 | Lincoln Park |
A | Geraghty | Lane | Wahrmann | Guarnizo | 9:01.34 | SLC Championships 2025 |
| 5 | St. Edward's |
A | Bryant | Perrin | Perrin | Taft | 9:25.31 | Benjamin Invite #1 2025 (Sanctioned) |
| 6 | Port St. Lucie |
A | Ajuccm | Wright | Nelson | Davis | 9:40.94 | Bulldog Invitational 2025 |