Mechanism specific biomarkers are used in ecotoxicology to identify classes of chemicals and to inform on their presence in the environment, but their use in signalling for adverse effects has been limited by a poor understanding of their associated links with health. In this study an experimental analysis was undertaken to investigate how induction or suppression of an estrogen-dependent biomarker, vitellogenin (VTG), related to health effects in fathead minnow (Pimephales promelas, FHM). Exposure to an oestrogen agonist, estradiol (29 and 60 ng/L), resulted in rapid induction of VTG (elevated plasma concentrations within 2 days of exposure) in male FHM that was subsequently slow to clear from the plasma (concentrations remained elevated 70 days after cessation of exposure). The induction of VTG to concentrations of 0.5 mg/mL, however, and its continued presence in the plasma were not associated with any overt adverse health effects to the males. In contrast, induction of higher concentrations of VTG (>1 mg/mL) in reproductively active FHM exposed to estrone (307 and 781 ng/L), were associated with impacts on male survival (>33% male mortality) and an inhibitory effect on egg production in females (>51% decrease in egg number). Exposure of reproductively active FHM to a chemical that disrupts estrogen biosynthesis (an aromatase inhibitor; fenarimol 497 μg/L) also reduced reproductive success (40% decrease in egg number), and this was associated with a reduction in plasma VTG concentrations in females (36% decrease). These findings show that high level induction or suppression (in females) of plasma VTG are associated with alterations in health status and reproductive fitness. VTG, therefore, has the potential to act as a health measure as well as a biomarker for exposure, for chemicals that alter the oestrogen signalling pathway.