Limestone Creek terminates in the Tennessee River at Arrowhead Landing, which is the southeasternmost point of Limestone County's Wheeler National Wildlife Refuge.

The land within this watershed is predominantly agricultural, but has experienced significant recent residential growth from the city of Huntsville.

Limestone Creek is a host to several species of fish, birds, and mammals.

Fish species include Micropterus coosae (redeye bass), Ambloplites rupestris (rock bass), several species of Lepomis (sunfish), Ictalurus punctatus (channel catfish), and Perca flavescens (yellow perch).

Multiple sightings of the American alligator (Alligator mississippiensis) have been reported, especially near the mouth of the river in Limestone Bay, which is the confluence of Limestone Creek with the Tennessee River.