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The impact of 17beta-estradiol (E2) and bisphenol A (BPA) on steroid hormone levels and gonad development in brown trout (Salmo trutta) was determined. Exposure took place from 0 to 63 days post-fertilisation (dpf) and gonad development was followed till 400 dpf. The onset of xenoestrogen metabolism was examined by measurements of whole body concentrations of bisphenol A (BPA) and its conjugation product bisphenol A glucuronic acid (BPAGA). Exposure to 500 ng E2/l led to an increase in E2 levels in the embryos and fry while 10 ng E2/l did not. Metabolic conversion of BPA to BPAGA began during the first weeks of embryonic development. Few consistent effects were found on the sex differentiation of the brown trout. Only one intersex fish (4.5%) was found among male fish at 400 dpf exposed to 500 ng E2/l. Females with male germ cells among the normally developing oocytes were observed in all groups (in up to 50% of the female fish, independently of exposure regime). The fact that exposure to 500 ng E2/l only caused subtle effects in a small number of individuals indicates that exposure during early life stages results in little to no induction of endocrine disruption in brown trout.

The kinetics of bisphenol A (BPA) were investigated in zebrafish (Danio rerio) exposed to 100 microg BPA/l. BPA uptake was measured during a 7-day period followed by an elimination phase of similar duration. After 2, 6, 12, 24, 48, 72, 120 and 168 h of uptake/elimination, fish were analysed for their content of BPA, bisphenol A glucuronic acid (BPAGA) and bisphenol A sulfate (BPAS). Within the first 24 h steady state levels of BPA, BPAGA and BPAS were reached and the total body concentrations were calculated to be 569, 12,600 and 39.9 ng/g fish, respectively. Elimination rates of the three compounds in zebrafish were estimated by fitting the data to a compartment model. An initial rapid elimination phase was observed for BPA and BPAS with total body half lives (T(1/2)) of <1.1 h and 30 min, followed by a slower second elimination phase with T(1/2) values of 139 and 71 h, respectively. Excretion of BPAGA occurred from a single compartment with a T(1/2) of 35 h. The steady state concentration of BPA and its metabolites were investigated in rainbow trout (Oncorhynchus mykiss) exposed to 100 microg BPA/l. The toxicokinetic parameters from zebrafish and rainbow trout were compared; including previously published data on the rainbow trout. The data indicate that the smaller estrogenic sensitivity observed for the zebrafish may be caused by a more rapid metabolism of BPA in the zebrafish liver.

The uptake, metabolism and excretion of the oestrogenic chemical bisphenol A (BPA) were studied in juvenile rainbow trout (Oncorhynchus mykiss). BPA was detectable in plasma, liver and muscle after 2 h of water exposure at 0.44 microM (100 microg BPA/l), and a steady state was reached within 12-24 h. The concentration of the glucuronidated degradation product in the plasma was about twice that of the parent compound. A plasma half life of BPA was calculated as 3.75 h following injection of the compound. The vitellogenin synthesis was measured in response to the BPA treatment, and a lag period of 5 and 7 days between injection of the compound and a significant vitellogenin response was observed for females and males, respectively. At the time of the vitellogenin response no BPA could be detected in the liver tissue from either male or female fish. These results indicate that fish briefly exposed to elevated levels of oestrogenic chemicals might develop a response several days later.

Extraction methods were developed for quantification of the xenoestrogens 4-tert.-octylphenol (tOP) and bisphenol A (BPA) in water and in liver and muscle tissue from the rainbow trout (Oncorhynchus mykiss). The extraction of tOP and BPA from tissue samples was carried out using microwave-assisted solvent extraction (MASE) followed by solid-phase extraction (SPE). Water samples were extracted using only SPE. For the quantification of tOP and BPA, liquid chromatography mass spectrometry (LC-MS) equipped with an atmospheric pressure chemical ionisation interface (APCI) was applied. The combined methods for tissue extraction allow the use of small sample amounts of liver or muscle (typically 1 g), low volumes of solvent (20 ml), and short extraction times (25 min). Limits of quantification of tOP in tissue samples were found to be approximately 10 ng/g in muscle and 50 ng/g in liver (both based on 1 g of fresh tissue). The corresponding values for BPA were approximately 50 ng/g in both muscle and liver tissue. In water, the limit of quantification for tOP and BPA was approximately 0.1 microg/l (based on 100 ml sample size).