| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note | 
|---|---|---|---|---|---|---|---|---|
| 3 |  Pine Ridge | 
		B | Quinones | Castro | Camacho | Rodriguez | NT | |
| 4 |  Mainland | 
		B | Parks | Rucker | Thompson | Ciferri | DQ | |
| 5 |  Deltona | 
		B | Ware | Lewis | Benjamin | Baldwin | NT | |
| 6 |  University (OC) | 
		B | Elder | Santos | Cruz | Carter | 3:55.00 | 
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note | 
|---|---|---|---|---|---|---|---|---|
| 1 |  Pine Ridge | 
		A | Alston | Genaro | Gavilanes | Hipster | 4:00.00 | |
| 2 |  University (OC) | 
		A | Andrews | Burns | Kwamina | Swint | 3:49.58 | 3:49.58 Deland HS Big D Relays 2024 | 
| 3 |  Deltona | 
		A | Castro | Pierre | Becerra | Haughton | 3:49.58 | 2024 George Janan Spruce Creek Invitational | 
| 4 |  Mainland | 
		A | Wilmore | Wood | Berry | Curry | 3:23.00 | |
| 5 |  Seabreeze | 
		A | Moore | Davis | Dayton | Lewis | 3:30.63 | FHSAA 3A District 6 | 
| 6 |  Trinity Christian (Deltona) | 
		A | Hudzik | Ponce-Arce | Thompson | Cruz | 4:08.82 | Geneva Developmental Opener 2025 |