| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Tradition Prep | A | Luongo | Luong | Ogilvie | Renteria | NT | |
| 2 | Centennial |
A | Garcia Guevera | Barber | Bryant Jr. | Kirkwood | 12:16.63 | Bill Wilson Invitational 2025 |
| 3 | Treasure Coast | A | Beausejour-Joseph | Borrows | Carroll | Lynch | NT | |
| 4 | Martin County |
A | Rey | White | Allison | Ziskinder | NT | |
| 5 | John Carroll |
A | Rohrborgh | Bradley | Sierra | Duffy | NT | |
| 6 | Fort Pierce Central |
A | Francois | Filsaine | Louissaint | Rivera | 10:00.00 | |
| 7 | Lincoln Park |
A | Guarnizo | Stefani | Geraghty | Lane | NT | |
| 8 | South Fork |
A | Quintero | Stoiber | Hartley | Robinson | NT | |
| 9 | Sebastian River |
A | Kross | Pankiewicz | Perez | Sanchez | 13:21.10 | Bill Wilson Invitational 2025 |
| 10 | Vero Beach |
A | Krough | Mitchell | Durham | Murphy Berek | 12:25.97 | Bill Wilson Invitational 2025 |