Anahola Stream is a perennial stream in northeast Kauaʻi, Hawaiʻi, that supports agricultural, domestic, and cultural uses within its drainage basin. Beginning in the late 19th century, Anahola streamflow was diverted by Makee Sugar Company at altitudes of 840 feet (upper intake) and 280 feet (lower intake) for irrigating sugarcane in the Keālia area. When sugarcane cultivation in the Keālia area ceased in 1988, part of the Makee Sugar Company’s surface-water collection system (Makee diversion system) in the Anahola drainage basin was abandoned. In an effort to better manage available surface-water resources, the State of Hawaiʻi Department of Hawaiian Home Lands is considering using the existing ditches in the Anahola Stream drainage basin to provide irrigation water for Native Hawaiian farmers in the area. To provide information needed for successful management of the surface-water resources, the U.S. Geological Survey investigated the availability and distribution of natural low flow in Anahola Stream and also collected low-flow data in Goldfish Stream, a stream that discharges into Kaneha Reservoir, which served as a major collection point for the Makee diversion system. Biological surveys of Anahola Stream were conducted as part of a study to determine the distribution of native and nonnative aquatic stream fauna. Results of the biological surveys indicated the presence of the following native aquatic species in Anahola Stream: ʻoʻopu ʻakupa (Sandwich Island sleeper) and ʻoʻopu naniha (Tear-drop goby) in the lower stream reaches surveyed; and ʻoʻopu nākea (Pacific river goby), ʻoʻopu nōpili (Stimpson’s goby), and ʻōpae kalaʻole (Mountain shrimp) in the middle and upper stream reaches surveyed. Nonnative aquatic species were found in all of the surveyed stream reaches along Anahola Stream. The availability and distribution of natural low flow were determined using a combination of discharge measurements made from February 2011 to May 2012 at low-flow partial-record and seepage-run stations established at locations of interest along study-area streams. Upstream of the upper intake, the estimated natural (undiverted) median flow in Anahola Stream is 2.7 million gallons per day, and the flow is expected to be greater than or equal to 0.97 million gallons per day 95 percent of the time. About 0.7 mile upstream of the lower intake and downstream from the confluence with Keaʻoʻopu Stream, the estimated natural (undiverted) median flow in Anahola Stream is 6.3 million gallons per day, and the flow is expected to be greater than or equal to 2.7 million gallons per day 95 percent of the time. In Goldfish Stream, about 0.4 mile upstream from the point of discharge into Kaneha Reservoir, the estimated natural median flow is 0.54 million gallons per day, and the flow is expected to be greater than or equal to 0.23 million gallons per day 95 percent of the time. The discharge estimates are representative of low-flow conditions in the study-area streams, and they are applicable to the base period (water years 1961–2011) over which they have been computed. The distribution of natural low flow in Anahola Stream was characterized through data collected during wet- and dry-season seepage runs. Seepage-run results show that Anahola Stream was generally a gaining stream under natural low-flow conditions. During the wet-season seepage run, Anahola Stream at the station located upstream of tributary Kaʻalula Stream had more than five times the flow that was measured upstream from the upper intake. The estimated total gain (including tributary inflow) in the 6.1-mile seepage-run reach was 6.97 million gallons per day; about 42 percent of that gain was groundwater discharge to the main channel of Anahola Stream. During the dry-season seepage run, about 34 percent of the estimated total gain of 3.93 million gallons per day in the same seepage-run reach was groundwater discharge to the main channel of Anahola Stream. A 15-percent seepage loss was estimated in a 0.3-mile reach downstream from the confluence of Anahola and Keaʻoʻopu Streams. The report summarizes scenarios that describe (1) surface-water availability under regulated conditions of Anahola Stream if the upper and lower intakes are restored in the future; and (2) amount of flow available for agricultural use at the upper intake under a variety of potential instream-flow standards that may be established by the State of Hawaiʻi for the protection of instream uses.