Background: Previous studies testing the association between the built environment and walking behavior have been largely cross-sectional and have yielded mixed results. This study reports on a natural experiment in which changes to the built environment were implemented at a university campus in Hong Kong. Longitudinal data on walking behaviors were collected using surveys, one before and one after changes to the built environment, to test the influence of changes to the built environment on walking behavior. Methods: Built environment data are from a university campus in Hong Kong, and include land use, campus bus services, pedestrian network, and population density data collected from campus maps, the university developmental office, and field surveys. Walking behavior data were collected at baseline in March 2012 (n = 198) and after changes to the built environment from the same cohort of subjects in December 2012 (n = 169) using a walking diary. Geographic information systems (GIS) was used to map walking routes and built environment variables, and compare each subject’s walking behaviors and built environment exposure before and after the changes to the built environment. Walking behavior outcomes were changes in: i) walking distance, ii) destination-oriented walking, and iii) walked altitude range. Multivariable linear regression models were used to test for associations between changes to the built environment and changes in walking behaviors. Results: Greater pedestrian network connectivity predicted longer walking distances and an increased likelihood of walking as a means of transportation. The increased use of recreational (vs. work) buildings, largely located at mid-range altitudes, as well as increased population density predicted greater walking distances.Having more bus services and a greater population density encouraged people to increase their walked altitude range. Conclusions: In this longitudinal study, changes to the built environment were associated with changes in walking behaviors. Use of GIS combined with walking diaries presents a practical method for mapping and measuring changes in the built environment and walking behaviors, respectively. Additional longitudinal studies can help clarify the relationships between the built environment and walking behaviors identified in this natural experiment