| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Lake Gibson |
A | Alonso | Brown | McBurrows | Whitehurst | NT | |
| 4 | DeSoto County |
A | Gonzalez | Brown | Johnson | Johnson | 55.24 | Lake Placid Invitational 2026 |
| 5 | Okeechobee |
A | Bowman | Ridley | Pineda | Fonseca | 59.30 | Lake Placid Invitational 2026 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Moore Haven |
A | Herrera | Myers | Hughes | Faison | 53.54 | Jensen Beach Invitational 2026 |
| 3 | LaBelle |
A | Deas | Williams | Sandoval | Jackson | DNF | Lake Placid Invitational 2026 |
| 4 | Sebring |
A | Gray | Greene | Wellington | Madison | 51.93 | Lake Placid Invitational 2026 |
| 5 | Avon Park |
A | Davi | Johnson | Destino | Watt | 52.26 | Avon Park Red Devil Invitational |
| 6 | Hardee |
A | Pierre | Silva | Manriquez | Nunez | DNS | Bob English Invitational 2026 |
| 7 | Lake Placid |
A | Dillon | Carlini-Smith | Johnson | Jacobs | 54.69 | Avon Park Red Devil Invitational |