| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Okeechobee |
A | Campbell | Summerford | Santos | Stewart | 53.39 | Lake Placid Invitational 2026 |
| 4 | Hardee |
A | Ohlinger | Gomez | Cherry | Watson | 46.58 | Lake Placid Invitational 2026 |
| 5 | Lake Placid |
A | Falls | Corby | Melgar | Abuelouf | 48.49 | Avon Park Red Devil Invitational |
| 6 | Four Corners |
A | Flax | Ubervil | Robinson | Ortiz | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Vanguard (LW) |
A | Cartright | Seymour | Khan | Lacossade | 45.72 | Avon Park Invitational |
| 2 | Moore Haven |
A | Fonseca | Adderley | Williams | Gunsby | 44.54 | Avon Park Red Devil Invitational |
| 3 | Avon Park |
A | Owens | Jones | Jones | Owens | 43.88 | Avon Park Red Devil Invitational |
| 4 | Sebring |
A | Brown | Austin | Volvaire | Hamilton | 42.61 | Lake Placid Invitational 2026 |
| 5 | Lake Gibson |
A | Hill | Jean-Bart | McBride Jr. | Dash | 42.67 | Lakeland City Meet |
| 6 | DeSoto County |
A | Hamm | Johnson | Grant | Wilkinson | 44.07 | Avon Park Invitational |
| 7 | LaBelle |
A | Hull | Thomas | Davis | Bradshaw | 44.89 | Lake Placid Invitational 2026 |
| 8 | Mulberry |
A | Nichols | OSteen | Cobbett | Townsend | 46.02 | Lake Region Sprint Meet 2026 |