Juvenile amphibians do not avoid potentially lethal levels of urea on soil substrate.

We examined the effects of a forest fertilizer (urea) on newly metamorphosed terrestrial amphibians (Western toads, Bufo boreas; Cascades frogs, Rana cascadae; long-toed salamanders, Ambystoma macrodactylum; and roughskin newts, Taricha granulosa). We examined avoidance behavior of Western toads and Cascades frogs on both paper towel and soil substrates dosed with urea (control and 100 kg N/ha and an additional treatment of 50 kg N/ha for Western toads on soil substrate) and avoidance behavior of long-toed salamanders on soil substrate dosed with urea. We further examined the survival and feeding behavior of all four species exposed to urea on soil substrate (100 kg N/ha) for 5 d. Juvenile Western toads and Cascades frogs avoided paper towels dosed with urea but did not avoid urea-dosed soil substrate. However, Western toads and Cascades frogs both suffered significant mortality when exposed to urea on a soil substrate for 5 d. Furthermore, after adjusting for weight, we found that urea-exposed juvenile Western toads and Cascades frogs consumed significantly fewer prey items (crickets) compared with nonexposed control animals. Long-toed salamanders did not discriminate against soil substrate dosed with urea, and neither long-toed salamanders nor roughskin newts died or reduced prey consumption as a result of urea exposure. Juvenile amphibians may not be able to detect and avoid harmful levels of urea fertilizer on a natural substrate. Furthermore, anthropogenic stressors such as urea fertilizer can significantly reduce the survival and prey consumption of juvenile amphibians. These effects are important to consider in light of possible threats to the conservation status of many amphibian species.

Combined effects of UV-B, nitrate, and low pH reduce the survival and activity level of larval cascades frogs (Rana cascadae).

We investigated interactions between low pH, high nitrate level, and ultraviolet-B (UV-B) light on the survival and activity level of larval Cascades frogs (Rana cascadae). We used a fully factorial experimental design, with pH levels of 5 and 7; initial "pulse" nitrate exposure levels of 0, 5, and 20 mg/L; and UV-B present or absent. After a 3-week laboratory exposure, we measured survival and activity level of the larvae. The experiment was repeated two times, in two separate years. Similar effects on survival and activity level were observed in both experiments. R. cascadae survival was not significantly reduced in treatments with individual factors alone (i.e., UV-B control, pH 5 control, or high nitrate level without pH or UV-B). However, in experiments from both years, survival and activity levels of larval R. cascadae were significantly reduced in the treatment with low pH, high nitrate, and UV-B together. In both years, analysis of variance (ANOVA) indicated that pH and nitrate had the greatest effect on survival and that UV-B and nitrate had the greatest effect on activity level. Additional effects were observed in the 1998 experiment on both survival and activity level. In 1998, UV radiation and the interaction term between pH and nitrate (pH x nitrate) had a significant effect on survival. Also in the 1998 experiment, activity level was significantly reduced in treatments at neutral pH with UV, at initial nitrate doses of 5 and 20 mg/L and at neutral pH without UV at an initial nitrate dose of 20 mg/L. We suggest that the adverse effects were due to the multiple stressors acting together.

Sediment toxicity and stormwater runoff in a contaminated receiving system: consideration of different bioassays in the laboratory and field.

Several field and laboratory assays were employed below an urban storm sewer outfall to define the relationship between stormwater runoff and contaminant effects. Specifically, two bioassays that measure feeding rate as a toxicological endpoint were employed in the field and in the laboratory, along with bioassays measuring survival and growth of test organisms. In 7 to 10 d in situ exposures, amphipod leaf disc processing, growth and survival were monitored. Different exposure scenarios were investigated by varying the mesh size (74 microns or 250 microns mesh) and method of deployment (water column, sediment surface, or containing sediment) of in situ exposure chambers. Hyalella azteca, Daphnia magna, and Pimephales promelas survival were monitored in 48 h in situ exposures. Feeding inhibition was investigated via enzyme inhibition of H. azteca and D. magna and via leaf disc processing measurements of the detritivore H. azteca. Additionally, we investigated the extent of phototoxicity at this site via field exposures in sun and shade and laboratory exposures with and without UV light. The measurement of detritivore leaf disc processing, and thus its usefulness as an endpoint, was hindered by individual variability in the amount of leaf consumed and by leaf weight gain during the summer field exposures. For D. magna, enzyme inhibition measured in a laboratory exposure did not reveal the toxicity observed in field exposures. For H. azteca, enzyme inhibition measured in the laboratory indicated toxicity similar to that observed in short term chronic in situ exposures. Enzyme inhibition also did not detect differences in toxicity due to variations in flow conditions. There were no statistically significant effects of any exposure on P. promelas survival or H. azteca growth, and there were no statistically significant effects due to mesh size or sun exposure. Survival of H. azteca was the most sensitive and the least variable endpoint. Effects on survival were noted in the same treatments over short-term chronic exposures in the laboratory and in situ. Significant differences in survival were noted due to the method of deployment under low flow conditions. In situ chambers containing sediment resulted in greater mortality in the 10 d low flow in situ experiments. Under high flow conditions, significant reductions in survival and leaf disc processing were noted under all methods of deployment at the two impacted sites over a 7 d exposure. Also under high flow conditions, significantly greater mortality of H. azteca was reported at the downstream field site when sediment was included in the chamber at deployment. These results suggest that significant toxicity at this site is due to accumulation of contaminants in the sediment and the mobilization of these contaminants during a storm event. In situ exposures detected toxicity not observed in laboratory exposures. These results suggest that a combination of laboratory and field bioassays is most useful in defining field effects.

Photo-induced toxicity of PAHs to Hyalella azteca and Chironomus tentans: effects of mixtures and behavior.

In the aquatic environment, polycyclic aromatic hydrocarbon (PAH) contamination can result from several anthropogenic sources such as petroleum runoff, industrial processes, and petroleum spills. When ultraviolet light (UV) is present at sufficient intensity, the acute toxicity of some PAHs to aquatic biota is greatly enhanced. This photo-induced toxicity of PAHs is directly influenced by the amount of PAH and by the level of UV intensity present in the aquatic environment. Thus, behavioral responses and habits that affect an aquatic organism's exposure to UV as well as exposure to PAHs can influence the extent to which damage due to photo-induced toxicity occurs. Experiments demonstrated the effects of photo-induced toxicity of anthracene and fluoranthene on the survival of two benthic macroinvertebrates, the midge Chironomus tentans and the freshwater amphipod Hyalella azteca. This study further investigated the survival and behavior of the test organisms in different substrates (no substrate, a sand monolayer, leaf discs, and sediment) with and without UV. The free-swimming, epibenthic H. azteca avoided the effects of photo-induced toxicity of PAHs to some extent by hiding in leaves when this substrate was available. Results emphasize the importance of organisms' behavior in affecting the photo-induced toxicity of PAHs in the aquatic environment.