| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Mount Gilead | A | Griffith | Rubio | Travis | Ammerman | 1:10.00 | |
| 3 | East Knox | A | Wengerd | Campos | Page | Heddix | 1:06.00 | |
| 4 | Danville | A | Stephens | Hackman | Keegan | Shaffer | 56.00 | |
| 5 | Fredericktown | A | Leist | Overholt | Overholt | Roddy | 58.49 | |
| 6 | Northmor | A | Ambrose | James | Keirns | Bosh | 1:02.60 | |
| 7 | Cardington-Lincoln | A | Greenawalt | Longsdorf | Moloney | Mooney | NT |