| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | All Saints' |
A | Cook | Cook | Fischer | Gunderson | 10:25.33 | FHSAA 1A Region 3 |
| 2 | Haines City |
A | Cote | Font Ruiz | Gonzalez | Mattos-Farias | 11:14.00 | |
| 3 | Hardee |
A | Alamia | Herrera | Gonzalez | Roberts | 11:14.42 | FHSAA 2A Region 3 |
| 4 | Sebring |
A | Meier | Brito | Allen | Ramos Negron | 11:25.54 | Hardee Invitational |
| 5 | Davenport |
A | Neal | Pollard | Sanchez | Morales-Orta | 11:30.03 | Kowboy Invitational |
| 6 | Winter Haven |
A | Rhodes | Huie | McPherson | Sakatos | 11:34.22 | 11:34.22 Polk County Championship 2023 |
| 7 | Frostproof |
A | Barajas | Sanchez | Gutierrez | Liboy | 11:52.87 | Frostproof - The Ridge Open 2025 |
| 8 | Tenoroc |
A | Henao | Chasteen | Suarez | Adams | DNS | 2025 Titan Invitational |