| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Powell Middle School | A | 57.92 | Challenger K8 Host, Fox Chapel, |
| 3 | Explorer K-8 School | A | 57.57 | DS Parrott Host Explorer K8, Wes |
| 4 | DS Parrot Middle School | A | 57.63 | DS Parrott Host Explorer K8, Wes |
| 5 | Classical Preparatory School | A | 1:02.76 | Zephyrhills Bulldog Invitational |