USBR 50 continues through Ohio, traversing 11 counties to the Market Street Bridge that connects Steubenville and Follansbee, West Virginia.

[13] In Xenia, USBR 50 shares a short segment of the Little Miami trail with State Bike Route 3.

The route begins at the Market Street Bridge at its crossing the Ohio River and connects to the Panhandle Trail and continues on into Pennsylvania.

USBR 50 mostly follows the Panhandle and Montour Trails and the Great Allegheny Passage as it crosses five counties to connect Colliers, West Virginia, with Frostburg, Maryland.

[14] From West Newton to Rockwood, it shares the Great Allegheny Passage with BicyclePA Route S.[15] From the Pennsylvania state line near Frostburg, USBR 50 continues along the Great Allegheny Passage and then follows the Chesapeake & Ohio Canal Towpath to the Georgetown neighborhood of Washington, D.C. U.S.

The first segment of USBR 50, incorporating the length of the C&O Canal Towpath in Maryland, was approved by the American Association of State Highway and Transportation Officials (AASHTO) on October 23, 2013.

[1] On May 29, 2014, AASHTO approved additional segments in Ohio and Washington, D.C., including the remaining 3.6 miles (5.8 km) of the C&O Towpath.

[9] AASHTO also approved a 2018 realignment of USBR 50 in Ohio, trimming 5 miles (8.0 km) from the route, passing through Hopedale and bypassing Alexandria.