The centralized Rio Seco Industrial Park (RSIP) tannery collective in the Southern Peruvian city of Arequipa releases untreated tannery wastewater into a proximal creek that is a tributary of the Chili River. As industrial leather tanning wastewater contains high concentrations of metal(loid)s, salts, dyes, and organics, this complex mixture could exert a myriad of toxicological effects on the surrounding ecosystem. The RSIP effluent was analyzed to quantify the acute ecotoxicity and ecotoxicological status of this untreated industrial wastewater at multiple trophic levels with the following bioindicators: the floating macrophyte Lemna minor, invertebrates Daphnia magna and Physa venustula, and the amphibian Xenopus laevis. A physicochemical characterization of the RSIP effluent revealed a highly contaminated waste stream. In addition to chromium (10.4 ± 0.4 mg/L) and other toxic metals, the water harbored extremely high concentrations of total dissolved solids (67,770 ± 15,600 mg/L), biochemical oxygen demand (1530 ± 290 mg/L) and total nitrogen (490 ± 10 mg/L). The toxicological responses of certain bioindicator species tested were evaluated after exposure to 0, 1.5, 3.0, and 4.5% untreated tannery wastewater blended with dechlorinated tap water. L. minor experienced a significant decrease in the number of fronds, wet weight, and dry weight at the lowest blended wastewater of 1.5%. Bioassays with D. magna showed the effect on neonatal mortality with a calculated LC50 of 1.1% for 48 h. Bioassays with P. venustula embryos showed high sensitivity to diluted effluent with complete mortality at 3.0% wastewater and above. Finally, X. laevis showed a high sensitivity to the dilutions with an LC50 of 1.6 for embryos and 1.8% for tadpoles. Although RSIP wastewater contains many potentially toxic components, chromium and total dissolved solids, with a major contribution from sodium, are best correlated with acute toxicity variables. This suggests that conductivity or analogous measurements could provide a rapid and affordable forensic tool to query acute ecosystem pressures. Collectively, the results indicate that the release of untreated tannery wastewater from RSIP can exert pronounced acute impacts across trophic levels with the need for treatment or dilution to below 1% of total flow. As the assays addressed acute toxicity, the necessary treatment and/or dilution to mitigate chronic effects is likely much lower. In conclusion, untreated RSIP tannery wastewaters represent an ecological risk to downstream aquatic ecosystems; this needs to be addressed to prevent current and future environmental consequences.