| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Jupiter Christian |
A | Gilliam | Brandon | FLORA | Evans | 3:59.30 | Pine Crest Panther Relays |
| 4 | Suncoast |
A | Mahan | Menardy | Monahan | DeStefano | 3:40.33 | Lincoln Knowles Invitational 2024 (FHSAA Sanctioned) |
| 5 | Martin County |
A | Allison | Treadwell | Ineus | St. Juste | 3:41.09 | Bulldog Invitational 2025 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Jupiter |
A | Brown | Peters | Saunders | Diaz | 3:31.97 | Clyde Harris Invitational at Boynton |
| 3 | Wellington |
A | Spence | Paul | Smith | Wroy | 3:27.76 | PBGHS Invitational 2025 |
| 4 | Benjamin School |
A | Sukhu | Arwood | Giles | McCorkle | 3:19.90 | FHSAA State Track & Field Championships (1A) |
| 5 | Centennial |
A | Johns | Enoch | Polynice | Jean-Pierre | 3:22.85 | Cobra Relays 2025 |
| 6 | Blanche Ely |
A | Jenkins | Desvallons | Goldsby | Hines | 3:28.97 | BCAA Track & Field Championship |
| 7 | Fort Pierce Central |
A | Ransfer | McMiller | Filsaine | Pascoe | 3:39.66 | 28th Annual Warrior Invitational 2024 |