Epidalea calamita embryos at Gosner stages 3 and 19, and larvae at Gosner stage 25, were exposed to different copper sulfate concentrations, ranging from 0.05 to 0.40 mg Cu L(-1), in 96-h acute toxicity tests. Embryonic and larval mortality, development, growth, and larval escape behavior were evaluated. LC(50) at 96 h obtained at Gosner stages 3, 19, and 25 were 0.22, 0.08, and 0.11 mg Cu L(-1), respectively. Embryonic and larval developments were delayed after 96 h of copper sulfate exposure. Growth was also affected and individuals in control treatments grew to twice the size of those exposed to copper concentrations over 0.2 mg Cu L(-1) during the experiments initiated at Gosner stage 19. Escape behavior was altered after 96 h of copper sulfate exposure; larvae showed shorter distances moved and abnormal displacement types. However, after 4 days of recovery process, most of the larvae showed normal escape behavior. For amphibians that develop in temporary wetlands, increased development time, lower size, and altered escape behavior might have repercussions on the number of individuals that can successfully complete metamorphosis and, consequently, on recruitment.
Habitat fragmentation and alteration have been described as the major factors relating to the decline observed in several amphibian species. In Spain, these processes are mainly associated with the large agricultural land area, the widespread use of toxic substances, and the agricultural practices used. All these aggressions affect wetlands and their watersheds, which are essential aquatic systems for many amphibian species. In an effort to discover where conservation and specific action plans could be more necessary, this study relates the amphibian richness in 28 representative Mediterranean wetlands to their characteristics and the activities carried out on their watershed. The required information considers diverse wetland variables. Our results show that watershed habitat heterogeneity is significantly positively correlated with amphibian richness, while the percentage of the watershed area devoted to olive cultivation is negatively correlated with it. The results allow us to classify wetlands according to their importance for amphibian conservation. Our results also show that 80% of the studied wetlands need specific action plans. Especially, more sustainable cultivation methods are required considering drastic changes in current agricultural practices. Finally, our data provides a baseline to understand future alterations due to agricultural practices.
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