Nitrogenous compounds enter the environment through various anthropogenic sources. Among these are nitrate (NO3-) and nitrite (NO2-) which can oxidize the heme moiety of hemoglobin and reduce the oxygen-carrying capacity of the molecule resulting in toxicity. Of the two anions, nitrite is more toxic. Hemoglobin levels are influenced by environmental conditions; thus, we hypothesized that hemoglobin levels would influence the toxicity of nitrite with low hemoglobin levels resulting in enhanced toxicity and high hemoglobin levels resulting in reduced toxicity. We tested this hypothesis by elevating hemoglobin levels with pyriproxyfen treatment and lowering hemoglobin levels using siRNA in Daphnia magna. Exposure to pyriproxyfen significantly elevated hemoglobin mRNA levels and induced copper coloration of the organisms, indicative of increased hemoglobin protein accumulation. siRNA treatment significantly reduced hemoglobin mRNA levels in both untreated and pyriproxyfen-treated organisms and attenuated copper coloration. Pyriproxyfen treatment increased the tolerance of daphnids to the acute toxicity of nitrite approximately 2-fold while siRNA treatment significantly decreased the tolerance of daphnids to nitrite toxicity. Results indicate that increased hemoglobin levels increase the tolerance of daphnids to nitrite toxicity which may serve to protect daphnids in environments subject to hemoglobin-elevating hypoxia or elevated temperatures.