Water quality monitoring is used to determine the impact of human activities on the environment. We evaluated water quality in the Água Boa stream, located within the municipality of Dourados, State of Mato Grosso do Sul, Brazil, by analyzing physico-chemical, chemical, and microbiological parameters, as well as chlorophyll concentrations. Five sets of water samples were collected between December 2012 and November 2013 from three locations within the stream. The results showed the presence of Escherichia coli and antibiotic-resistant Pseudomonas spp. strains and high concentrations of organic matter (total dissolved solids), inorganic species (Mg, Ca, and Fe), and agrochemical residues (thiamethoxam). The main stream water contaminants are derived from urban, industrial, and agricultural activities within the watershed. Given the presence of contaminants, it is important that such findings are disseminated in order to highlight the risks that contact with this water may pose to human health. To preserve the environment and improve site conditions, people would need to participate by demanding that normative national and international standards be respected and that the situation be supervised by the competent governmental agencies; this would make it possible to reverse or minimize contamination problems within the Água Boa stream.