| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Northmor JH | A | Short | Young | Frank | Hines | 55.29 | Northmor MS Meet |
| 4 | Loudonville JH | A | Caley | Caley | England | Ross-Patrick | 50.64 | Triway JH Invite |
| 5 | Centerburg MS | A | Jones | Nelson | Rhodebeck | Rice | 51.05 | CSG MS April 28 2025 |
| 6 | Danville | A | Sampson | Sampson | Byers | Hackman | 53.54 |