Waterloo – St. Jacobs Railway, Heritage railway in Waterloo Region, Canada.

The Waterloo–St. Jacobs Railway was a heritage railway west of Toronto that ran between Waterloo and St. Jacobs, Ontario, from 1997 to 1999. It used two 1950s diesel locomotives from Canadian National Railways, named Spirit of St. Jacobs and Pride of Waterloo, with passenger cars in the same paint scheme. Excursions had three stops, and riders could return on a later train.

The Waterloo–St. Jacobs Railway ran between Waterloo and St. Jacobs, Ontario, from 1997 to 1999, using two 1950s diesel locomotives from Canadian National Railways named Spirit of St. Jacobs and Pride of Waterloo. It ceased operation in 1999 because of maintenance costs. In May 2007 the Southern Ontario Locomotive Restoration Society won City of Waterloo approval for a successor tourist train, the Waterloo Central Railway.

The line ran between Waterloo and St. Jacobs with three stops, and a ticket let riders get off and return on a later train. Passengers travelled past farms of the Old Order Mennonites and could spend time in the village of St. Jacobs.

The train services operate on a seasonal schedule with service running on specific days of the week. It is helpful to check the operating calendar in advance to plan your visit and confirm which stops and destinations are available when you intend to travel.

The two 1950s diesel locomotives, named Spirit of St. Jacobs and Pride of Waterloo, carried a paint scheme devised for the line. After the railway closed in 1999, the West Coast Railway Association bought the diesels and three passenger cars in late 2006; by 2008 the locomotives and one car were still stored in Via Rail's Mimico yard.