| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Tampa Bay HEAT |
A | Forsyth | Mueller | Bailey | Davis | NT | Tenoroc High School Invitational at SEU |
| 3 | South Sumter |
A | Howell | Lewis | Morgan | Smothers | 50.00 | |
| 4 | Circle Christian |
A | Sill | Lao | Mauck | Keller | 48.62 | Charlie Harris Relay |
| 5 | Seven Rivers Christian |
A | Bailey | Vann | Novack | Kingrey | 48.83 | Citrus Round Up Championship |
| 6 | First Academy (Leesburg) |
A | Williams | Edwards | Simmons | Ogden | 55.00 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Crystal River |
A | Johnson | Jenkins | Wellington | George | NT | 2026 Kiwanis Invitational |
| 3 | Dunnellon |
A | Devine | Mahns | Walker | Oldham | NT | Wildwood Open |
| 4 | Lake Minneola |
A | Verghese | Browne | Reams | Robinson | NT | FHSAA 4A Region 1 |
| 5 | Wildwood |
A | James | Fredericks | Benitez-Sanquiche | Smith | NT | |
| 6 | Citrus |
A | Brown | Macias | Cobb | Takett | NT | FHSAA 3A District 5 |
| 7 | Lecanto |
A | Brashear | Sheppard | Barbierri | Curylo | NT |