The 14,882-acre (60 km2) forest is located in North Central Florida, along the banks of the Withlacoochee and Suwannee rivers.

[1] According to the Florida Department of Agriculture's official website, Twin Rivers State Forest is composed of 15 noncontiguous tracts.

The former is across the river from Madison Blue Spring State Park, and the latter is also along SR 6's overlap with Hamilton CR 143.

The Hardee Springs Tract and Westwood East Tract are located on the opposite side of the Withlacoochee West Conservation Area, and the Westwood West Tract is located north of this conservation area on the opposite side of the river.

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