Roundup formulations are herbicides whose active principle is glyphosate. However, these formulations are potentially more toxic to non-target organisms than pure glyphosate. This study aimed to evaluate and compare the toxic potential of the Roundup formulations through histological alterations in fish. Thus, males and females of the neotropical fish species Jenynsia multidentata (Jenyns, 1842) were exposed for 24 or 96 h to the Roundup Original® (RO), Roundup Transorb® (RT) or Roundup WG® (RWG) formulations, at a fixed concentration of 0.5 mg/L of glyphosate. This concentration is close to the maximum glyphosate limits found in the environment and is non-lethal to J. multidentata. The three formulations caused histological damage to the liver, gills and brain of J. multidentata, which increased over the exposure time. Differences in the histological alterations between females and males were observed in the liver and brain. Females were more tolerant to RO and RT than RWG. Males did not exhibit these differences in sensitiveness with formulations. The RWG caused more damage in the liver and gills and RT in the brain. Overall, there were differences in the toxicity of RO, RT and RWG and the toxic effect was presented through histological damage, reinforcing the usefulness of histological biomarkers for Roundup® toxicity. The comparison of the toxic potential of glyphosate-based herbicides is important because it could give support to the governmental organizations to set protective rules for the water ecosystems and human health, as well as to reduce the use of highly toxic formulations in agriculture.