| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Lake Placid |
A | Melgar | Leon | Zepeda | Abuelouf | 3:55.68 | Avon Park Red Devil Invitational |
| 3 | DeSoto County |
A | Hernandez | Tanner | Wilkinson | Shine | 3:40.63 | Avon Park Chet Brojek Track Invitational |
| 4 | Clewiston |
A | Exilhomme | mullings | Graham | Bryant | 3:55.42 | Benjamin Meet #2 2026 (Approved Qualifier) |
| 5 | Moore Haven |
A | Zefra | Gunsby | Fonseca | Adderley | 4:16.75 | 62nd Eagle Invitational |