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• Fe, Mn, Ni exceeded limits by MINAM/WHO in Andes mountain range Ancash-Peru waters • Mitotic index decreased with increasing concentration of Río Negro sub-basins' waters • Genotoxicity with frequency of micronuclei was highest in the upper sub-basin waters • Allium cepa and Vicia faba tests monitored toxicity in Andes mountain range waters • MN indicated a possible genotoxic risk to environmental health from glacial waters Climate change and global warming cause deglaciation and leaching of the Andes Mountain range, generating mixtures of metals carried by the waters of the Negro River in Ancash–Peru, which can negatively affect human, animal, plant, and environmental health, causing genotoxicity and cytotoxicity that can be evidenced by biomarkers such as micronucleus (MN) frequency and mitotic index (MI). Constant environmental monitoring maintains good health, and water quality assessment is essential for homeostasis. The objective was to evaluate the cytotoxicity and genotoxicity of water from the sub–basins of the Negro River in Ancash, Peru, using biological models of Allium cepa and Vicia faba . In the methodology, the waters of the Negro River of glacial origin from the Andes Mountain range, according to the collection altitude, were called upper, middle and lower sub–basins (9°38′40 S and 77°23′12 W), serial dilutions of concentrations of 100%, 50%, 40%, 30%, 20%, 10% and 2% (v/v), a negative control H 2 O and a positive control As 2 O 3 10 mg/L; with three independent laboratory repetitions in each sub-basin, biological model and concentration. Then, in Allium cepa and Vicia faba , water quality was evaluated using cytotoxicity tests with MI and the inhibitory concentration 50 (IC50), and genotoxicity tests with MN frequency. The results showed a possible cytotoxic effect of the waters, in the upper, middle, and lower sub–basins, by inhibiting the root growth of Allium cepa with IC50 of 4.37%, 12.42%, 17.32%, respectively; and Vicia faba 16.61%, 17.14%, 63.34% (p ≤ 0.05). In the evaluation of the MI, the upper and middle sub–basins showed a cytotoxic effect at dilutions of 100% – 50% by inhibiting the MI in both models and showed a cytostatic effect at dilutions of 40% – 20% by decreasing the values of the MI in both models. In Allium cepa , a decrease in the MI of 5.75%, 5.06%, 4.79% (upper sub–basin) and 6.18%, 5.89%, 5.34% (middle sub–basin) was observed, compared to the negative control (p ≤ 0.05). In the lower sub–basin waters, a decrease in MI was observed in dilutions of 100% – 2% in both models compared to the negative control (p ≤ 0.05). Furthermore, they showed a possible genotoxic effect of the waters, in the upper sub–basin, in the 40% – 2% dilutions, there was a significant frequency of MN compared to the negative control, with the Allium cepa model being more sensitive, and in 40%, there was a higher frequency of MN of 1.07% than As 2 0 3 . In the middle and lower sub–basins, there was a higher frequency of MN in Allium cepa compared to the negative control (p ≤ 0.05), and in the 40% dilution for the middle sub–basin, there was a higher frequency of MN of 0.67% than As 2 0 3 . In the lower sub–basin, in 100% – 40%, there was a higher frequency of MN of 0.51%, 0.49%, and 0.47% than As 2 0 3 . It was concluded that the Allium cepa and Vicia faba models were very sensitive to assess water quality; a greater cytotoxic and genotoxic effect was observed in the waters of the upper sub–basin, followed by the middle and lower sub–basins, and there was a greater toxic effect with the highest concentrations of water. The water report showed a mixture of ten heavy metals and twenty–three non–heavy metals; these waters were sampled in a single environmental campaign without temporal replication. Therefore, the waters of the sub-basins of the Negro River in Ancash-Peru could cause significant cytotoxicity, cytostaticity, and genotoxicity in biological models.