| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Lincoln Park |
A | Chagani | Eshleman | Kapsis | Holland | NT | |
| 4 | Martin County |
A | Dixon | Fucile | Parks | Ineus | 47.47 | John I Leonard Invitational |
| 5 | Fort Pierce Westwood |
A | Boone | Ford | Gardner | Lynch | NT | |
| 6 | Treasure Coast | A | Harper | Isaac | Jones | Reed | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Sebastian River |
A | Cummings | Hogue | Derice | Mewborn | 44.58 | Bill Wilson Invitational 2025 |
| 3 | Tradition Prep | A | Richardson | Atwell | Haye | Drummond | DNF | Bulldog Invitational 2025 |
| 4 | Fort Pierce Central |
A | Ransfer | Hills | Reynolds | Saint-Jean | 40.00 | |
| 5 | Centennial |
A | Johns | Dorelus Jr. | Polynice | Jean-Pierre | 40.96 | Bill Wilson Invitational 2025 |
| 6 | Vero Beach |
A | Neal | Jacobs | Jansky | Stinson | 44.35 | Bill Wilson Invitational 2025 |
| 7 | John Carroll |
A | Monaco | Coleman | Smith | Bradley | 45.29 | Benjamin Invite #1 2025 (Sanctioned) |