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The South Fork Little River (SFLR) and the North Fork Little River (NFLR) are two major headwater tributaries that flow into the Little River just south of Hopkinsville, Kentucky. Both tributaries are included in those water bodies in Kentucky and across the Nation that have been reported with declining water quality. Each tributary has been listed by the Kentucky Energy and Environment Cabinet—Kentucky Division of Water in the 303(d) List of Waters for Kentucky Report to Congress as impaired by nutrients, pathogens, and sediment for contact recreation from point and nonpoint sources since 2002. In 2009, the Kentucky Energy and Environment Cabinet—Kentucky Division of Water developed a pathogen total maximum daily load (TMDL) for the Little River Basin including the SFLR and NFLR Basins. Future nutrient and suspended-sediment TMDLs are planned once nutrient criteria and suspended-sediment protocols have been developed for Kentucky. In this study, different approaches were used to identify potential sources of fecal-indicator bacteria (FIB), nitrate, and suspended sediment; to inform the TMDL process; and to aid in the implementation of effective watershed-management activities. The main focus of source identification was in the SFLR Basin. To begin understanding the potential sources of fecal contamination, samples were collected at 19 sites for densities of FIB (E. coli) in water and fluvial sediment and at 11 sites for Bacteroidales genetic markers (General AllBac, human HF183, ruminant BoBac, canid BacCan, and waterfowl GFD) during the recreational season (May through October) in 2013 and 2014. Results indicated 34 percent of all E. coli water samples (n=227 samples) did not meet the U.S. Environmental Protection Agency 2012 recommended national criteria for primary recreational waters. No criterion currently exists for E. coli in fluvial sediment. By use of the Spearman’s rank correlation test, densities of FIB in fluvial sediments were observed to have a statistically significant positive correlation with drainage area. As drainage area increased, so did the densities of FIB in the fluvial sediments. There was no statistically significant correlation between drainage area and FIB in water. The human-associated marker (HF183) was found above the detection limit in 26 percent of the samples (n=120 samples); a higher proportion of positive samples was in the NFLR Basin. The ruminant-associated marker (BoBac) was above the detection limit in 65 percent of samples; a higher proportion of positive samples was in the headwaters of the SFLR Basin. Nutrient yields differed between the SFLR and NFLR Basins. Comparatively, the SFLR Basin produced the largest estimated mean yields of total nitrogen (16,000 pounds per year per square mile (lb/yr/mi2) and nitrite plus nitrate nitrogen (12,500 lb/yr/mi2), and the NFLR Basin produced the largest estimated mean yields of ammonia plus organic nitrogen (4,700 lb/yr/mi2), total phosphorus (1,100 lb/yr/mi2), and orthophosphorus (590 lb/yr/mi2). Nitrate sources in surface water were assessed in both basins using dual-nitrate isotope (nitrogen and oxygen) ratios. Data from the different land uses in the SFLR Basin showed differences in nitrate concentrations and overlapping, but moderately distinct, isotopic signatures. Predominantly forested sites consistently had low nitrate concentrations (median = 0.233 milligrams per liter) with minimal variability, and agricultural sites had the highest nitrate concentrations (median = 7.55 milligrams per liter) with the greatest variability. The median nitrate concentration for sites with mixed land use was 2.66 milligrams per liter. Dual-isotope data for forested sites plotted within ranges characteristic of soil-derived nitrate with possible but minimal influence from recycled atmospheric nitrate. Ranges of dual-isotope data for sites with agricultural and mixed land uses were characteristic of possible mixtures of chemical fertilizer, soil-derived nitrate, and manure and septic wastes. In the NFLR Basin, a positive linear relation was observed between nitrate concentrations and nitrogen isotope ratios (δNNO3) (R 2=0.56; p-value <0.001) that potentially suggests the NFLR Basin has a higher proportion of δNNO3-enriched sources, such as manure and sewage. However, mixing of other nitrate-derived sources cannot be excluded, because many values of δNNO3 and concentrations of nitrate showed minimal variation and plotted within dual-nitrate isotope ranges characteristic of fertilizer and soil-derived nitrate sources. 2 Multiple-Source Tracking: Pathogens, Nutrients, and Sediment, Upper Little River Basin, Kentucky, Water Years 2013–14 A sediment-fingerprinting approach was used to quantify the relative contribution of four upland sources in the SFLR Basin (agricultural, pasture, riparian/forest, and streambank) to understand how land management affects suspended-sediment concentration. Carbon isotope ratios (δ13C), together with calcium and carbon concentrations, were the best indicators of sediment source; the uncertainty was less than 11 percent. Fine-sediment samples collected at the SFLR Basin outlet indicated streambanks as the largest source of the fine sediment to the stream followed by cropland and riparian/forestsource areas, respectively; pasture was a minor contributing source. Streambanks and cropland were essentially equal contributors of fine sediment at the NFLR Basin outlet.

Multiple-source tracking: Investigating sources of pathogens, nutrients, and sediment in the Upper Little River Basin, Kentucky, water years 2013–14