The main goal of the present study was to show whether testicular cells of rainbow trout (Oncorhynchus mykiss Walbaum) either hormonally manipulated (XX males) or produced by using gamma irradiation and pressure shock (YY males, "supermales") are able to aromatize androgens into estrogens compared with the control (XY males). The expression of aromatase gene at the level of the protein and its presence in testicular tissue was investigated by means of immunohistochemistry and Western blot analysis, respectively. The positive staining for aromatase was detected in testicular cells of all trout and in efferent duct cells of XY and YY males. However, the staining intensity varied among particular trout, being strong in YY males, moderate in XY males, and weak in XX trout. It was confirmed by quantitative image analysis in which the staining intensity was expressed as relative optical density (ROD) of diaminobenzidine deposits. Significant differences were found between XY and YY trout ((**)p<0.01) and XY and XX trout ((*)p<0.05). Such differences could reflect various levels of estrogens, possibly dependent on the genetic background of the trout studied. It seems likely that differential expression of the enzyme, especially that of weak or strong intensity, causes some alterations in testicular morphology of homogametic trout. Additionally, the results indicate that an imbalance in sex hormone biosynthesis may provoke the functional alterations in testes of YY males, and, in consequence, negatively affect the fertility of "supermales".