| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Lake Placid |
A | Washington | Walton | Charite | Anderson | DNS | |
| 4 | Avon Park |
A | Timmons | Payne | Watkins | Burgess Jr. | 3:56.65 | Frostproof - The Ridge Open 2025 |
| 5 | Clewiston |
A | Mullings | mullings | Moody | Mason | 3:57.40 | 65th Annual Edison Relays |