The objective of this 3-year study was to characterize benthic communities and physical habitat in an agricultural stream in the mid-west area of the United States (Big Bureau Creek, Illinois). Concurrent basic water quality parameters and seven nutrients were measured in the water column. Sediment measurements from depositional areas were conducted for bifenthrin, Total Organic Carbon, grain size, polychlorinated biphenyls (PCBs) and eight metals. All parameters were measured at 12 sites annually during the late summer for a 3-year period (2014, 2015 and 2016). Univariate regressions, stepwise multiple regressions and canonical correlation statistical analyses were used to determine the relationship between various benthic metrics (i.e., taxa richness and abundance) and all the measured parameters for the 3-year database. Benthic communities comprising 108-110 taxa were collected annually, and were generally dominated by sensitive caddisflies and mayflies. These communities were rated as good to exceptional using the Ohio Invertebrate Community Index. Physical habitat for the various sites was rated as good using the Ohio Qualitative Habitat Evaluation Index, thus suggesting that habitat is not a significant stressor that would likely impact resident benthic communities. Based on a comparison of measured in-stream total nitrogen and total phosphorus concentrations and criterion value exceedances, it appears that the in-stream nutrient concentrations could be potentially stressful to resident benthic biota. Metal concentrations were below established NOAA Threshold Effects Levels at all sites. Measured PCB concentrations were below levels of detection at all sites. Toxic units' (TUs) calculations based on using sensitive laboratory strains of Hyalella were less than 0.1 for bifenthrin, thus suggesting that bifenthrin sediment toxicity was unlikely. Thirty significant relationships reported between benthic metrics and the various environmental variables based on the 3-year database were as follows: 12 relationships with habitat metrics, 8 relationships with metals, 7 relationships with nutrients, 2 relationships with bifenthrin and 1 relationship with sediment characteristics. Relative habitat quality, habitat preferences, and ecological competition appear to be factors that shape the characteristics of the benthic communities of Big Bureau Creek, rather than stress imposed by toxicants (bifenthrin and metals), nutrients, or sediment characteristics. Complex relationships between benthic metrics and environmental variables are discussed.