Streams in the Ozark Plateaus Physiographic Province occupy narrow, steep-sided valleys bordered in places by sheer limestone bluffs. Stream channels typically have clear, well-defined riffles and pools, and substrate consisting of gravel, cobble, pebble, boulder, sand and bedrock. This physiographic province is noted for its numerous springs, the largest of which emerge along the major streams and provide an abundant and reliable source of cool water for a considerable distance.

In Missouri, the Ozark shiner is reported to inhabit sections of clear streams with high gradients and permanent strong flow which occurs most abundantly near riffles in a slight to moderate current, over a firm, silt-free bottom. The species occasionally occurs in sections of large creeks but primarily lives in rivers. In the Black River in Missouri, biologists report the Ozark shiner from riffles with slower flowing water in association with wedgespot and carmine shiners. Ozark shiners are also reported from the headwater and tailwater zones, transitional between pools and riffles in association with whitetail, popeye, wedgespot and carmine shiners.

In Arkansas, biologists have found the Ozark shiner in sections of large creeks and small-to-medium rivers where the water is clear and cool to cold. During a pre-impoundment study in the Beaver Lake drainage, Ozark shiners occurred in pools of upstream sections with 0.95 to 3.79 meters per kilometer (5 to 20 feet per mile) gradient, whereas it was rare in downstream sections where gradients were less than 0.95 meters per kilometer (5 feet per mile). However, Ozark shiners were not found it in smaller tributaries of the White River. Other studies reported the Ozark shiner in deep pools, with some occurring over sand and gravel bottoms. Biologists conducting surveys during the 1970s in Jane’s and Piney creeks, in Arkansas, reported collecting the Ozark shiner only from pool areas. Others have reported this shiner inhabits sections of medium-to-large, clean streams with high gradient and permanent, strong flow; most common near riffles in slight to moderate current over firm, silt-free bottom. The Ozark shiner as a midwater schooling fish species, which prefers high-gradient stream sections of clear, upland medium streams to large rivers, usually occur just below riffles in slight-to-moderate current over gravel, cobble or sand bottoms. It can be found in deep, flowing pools or runs in the mainstem of the Spring River, in Arkansas, but also near the confluences of Myatt and Janes creeks. Larger river segments, like the Spring River mainstem, appear to support the greatest abundance of the Ozark shiner.

The Ozark shiner is a small, slender fish. Length of adults commonly 45.7 to 63.5 millimeters (1.8 to 2.5 inches) to a maximum of 71.1 millimeters (2.8 inches).  

Coloration of the dorsum is pale yellow, sides are silver with a dusky stripe that fades out toward head, and belly is silvery-white. The dorsal fin has a small dusky blotch near the base on the first two to three rays; all other fin membranes are often dusted with fine dark speckles in adults.

Limited life history information exists for many species belonging to the family Leuciscidae in North America due to difficulty in identification caused by their small size and little economic or cultural value. However, it is known that all members of this family lay eggs, and most do not guard them. 

There are no known estimates of survival nor growth for the Ozark shiner. However, its sister species, the mirror shiner, has a maximum life span of four years and was estimated to grow 1.05 millimeters a month (0.04 inches) in North Carolina streams. 

In a sequence analysis of the mitochondrial cytochrome b gene, the Ozark shiner was discovered as sister to a clade composed of the ghost shiner (Notropis buchanani) and mimic shiner (Notropis volucellus).

The channel shiner (Notropis wickliffi), sawfin shiners (Notropis species), mirror shiner (Notropis spectrunculus), tennessee shiner (Notropis leuciodus) and silverband shiner (Notropis shumardi) also have been included within this species complex, of which the closest related species to the Ozark shiner is the mirror shiner.