A study was conducted with the specie Gambusia punctata of Filé river in Santiago de Cuba with the aim of establishing reference values regarding histology and threshold values of heavy metals. Three target organs were tested: liver, branchiae and brain. Different sex adult individuals 282 were selected as class intervals; they measured between 2.1-3.0 and 3.1-4.0cm of total length. For the histological analysis the samples were treated as a pool, the organs were fixed with formaldehyde 10% and processed with an H-E dye so as to evaluate the morphological and tissue damage. The metals involved were: copper, zinc, cadmium and lead. They were treated with humidity and quantified by Inductively Coupled Plasma Spectrometry (ICP-AES). According to the histological analysis, no damages were found in the tested tissues. No metal levels were found above the threshold value in brain, as well as cadmium and lead in the liver. At the branchiae all the analyzed metals were similar to the threshold values where according to ANOVA there were not statistically significant differences (p<0.05) between the established class intervals. Conclusion: In spite that, this study is one of the first of this kind, with this specie in Cuba, the histological analysis, and the levels of the bioaccumulated heavy metals in the organs of interest allow to use this specie as reference for the studies of environmental aquatic ecotoxicology the histological analysis, toxicology.