Steroidal hormones produced by humans and animals are constantly being excreted into the environment. It has been demonstrated that sewage effluent discharged to surface water contains natural estrogens and synthetic estrogenic chemicals. As estrogen levels continuously increase in the aquatic environment, it is very important to have a detailed understanding of estrogens' effects on fish. In the present study, juvenile rainbow trout (Oncorhynchus mykiss) were exposed to 17 beta-estradiol (E2) for 3 weeks and the effects of E2 on rainbow trout livers were investigated at the molecular level using Fourier transform infrared spectroscopy. The results revealed that E2 induced significant alterations in the liver tissues. A decrease in glycogen levels and protein concentration, and an increase in both the population of hepatic lipids, especially triglycerides, as well as the relative content of nucleic acids was observed in the E2 treated liver. In addition, a decrease in the membrane fluidity and an increase in lipid order were found in the cells of treated samples. In order to compare the effect of E2 with that of NP at molecular level, the fish were also treated with an estrogenic compound, nonylphenol (NP). The NP-treated fish liver spectra were found to be quite similar to those of E2-treated fish confirming that NP mimics the effect of E2 in immature rainbow trout. (c) 2005 Elsevier B.V. All rights reserved.