Area ca. 56,000 acres. including about 25 miles of the Niobrara River, in the heart of the transition zone between western coniferous and eastern deciduous forest types. There is a Niobrara Valley Preserve bird checklist of 186 species, including a list of 75 definite and 30 more possible breeding species. There are also local checklists of other major animal and plant groups. The preserve has been identified as a Globally Important Bird Area by the American Bird Conservancy. Among the breeding birds of special interest are the eastern and western forms that hybridize here, such as the Baltimore and Bullock’s orioles, the lazuli and indigo buntings, and the rose-breasted and black-headed grosbeaks. The eastern and western wood-pewees may also hybridize here. Both Common Poorwill and Whip-poor-will can be heard here in the early evenings.
Two trails radiate out from the headquarters that pass through several forest types and the Sandhills prairie vegetation on the uplands. Each trail has a short loop and a long loop; the northern one is somewhat longer (three miles) and steeper. In the summer of 2012 a large fire burned through the preserve, especially on the north side of the Niobrara River. The preserve lies within the Niobrara National Scenic River district. It extends for 76 miles east from near Valentine downstream to join the Missouri River National Scenic River District, and is a popular canoeing destination.