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Aquatic organisms are exposed to a multitude of contaminants and to fully understand the impact of multiple stressors on fish populations, we must first understand the mechanism of action for each toxicant and how the combined effects manifest at the level of the individual. 17α-ethinylestradiol (EE2) has been known to cause adverse reproductive effects including reduced fecundity and fertility, intersex and skewed sex ratios in fish by mimicking naturally produced estrogen at low concentrations. Ammonia can cause adverse reproductive and mortality effects in individual fish through effects or damage to the central nervous system. Both EE2 and ammonia are found in most municipal effluents in various concentrations. A flow-through diluter system was used to test the individual effects of these two contaminants at their respective no observable adverse effect concentration (NOAEC) as well as their combined effects on fathead minnow, ( Pimephales promelas ) reproduction in a mixture exposure. While neither contaminant nor their mixture altered reproduction in terms of fecundity, their mixture resulted in significant fathead minnow mortality during a 21 d exposure. This study demonstrated the need to consider mixture effects when assessing risk for toxicity testing with multiple stressors. [ABSTRACT FROM AUTHOR]

Abstract: Ammonia can cause adverse reproductive and mortality effects in individual fish by interacting with the central nervous system. The last published study that assessed the effects of ammonia on fathead minnow reproduction was a lifecycle study conducted in 1986. Our study''s main goal was to re-evaluate ammonia toxicity on fathead minnow Pimephales promelas reproduction using a 20-day fecundity flow-through diluter method. Flow-through diluter systems have been used by regulatory agencies, such as the U.S. Environmental Protection Agency, in the past as an effective way to estimate acceptable levels of contaminants. There was a significant difference in cumulative egg production among treatments (ANOVA; F=10.167, p≤0.01, df=3). All three concentrations of ammonia tested in this study significantly reduced fecundity after 20days of exposure (Dunnett''s, p≤0.05 for each treatment). The lowest un-ionized ammonia concentration (0.06mg/L at a pH of 7.3 and temperature of 25.1°C) tested during this study resulted in a 29% decrease in cumulative fecundity. Because all tested ammonia concentrations caused an effect on P. promelas reproduction, the no effect concentration was estimated to be 0.025mg/L un-ionized ammonia (2.19mg/L total ammonia–nitrogen). This estimate was determined using the U.S. EPA Toxicity Relationship Analysis Program to calculate the 10% effect concentration of ammonia on P. promelas reproduction. This value is much lower than the previous reported no effect concentration on P. promelas reproduction (0.37mg/L un-ionized ammonia or 6.43mg/L total ammonia–nitrogen) as determined from the 1986 study, which was used to determine the ammonia water quality criteria by the U.S. Environmental Protection Agency. Our results should be considered in the next revision of water quality criteria. [Copyright &y& Elsevier]