| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 4 | East Lee County |
A | Georges | Rogers | Tobler | Brooks | DNS | 2024 South Fort Myers Wolfpack Invitational Sanctioned |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Estero |
A | Cavazos | Philipps | Hall | Small | 49.27 | 2024 South Fort Myers Wolfpack Invitational Sanctioned |
| 2 | Ida Baker |
A | Long | Bayley | Neely | Olmstead | 44.20 | Edison Relays 2024 (Approved Sanctioned Meet) |
| 3 | Mariner |
A | Davidson | Carter | Clark | Wilson | 43.22 | 2024 South Fort Myers Wolfpack Invitational Sanctioned |
| 4 | Dunbar |
A | Tyre | Green | Perez | Martinez | 41.20 | FHSAA 3A Region 3 |
| 5 | South Fort Myers |
A | bernadel | Raphael | Means | Michel | 42.49 | FHSAA 3A Region 3 |
| 6 | Gateway (Fort Myers) |
A | Fomby | Gourley | Starr-Howard | Louis | 44.55 | 44.55 59th ANNUAL EAGLE INVITATIONAL |
| 7 | Cypress Lake |
A | Johnson | Mckay | Johnson | Angarita | DNS | Guy Thomas Memorial Classic (FHSAA & NFHS Sanctioned) |
| 8 | Gulf Coast |
A | Akolo | Lormil | Ibrahim | Rivers | 45.00 |