We used a phylogenetically based comparative approach to evaluate the potential for physiological studies to reveal patterns of diversity in traits related to susceptibility to an environmental stressor, the trace metal cadmium (Cd). Physiological traits related to Cd bioaccumulation, compartmentalization, and ultimately susceptibility were measured in 21 aquatic insect species representing the orders Ephemeroptera, Plecoptera, and Trichoptera. We mapped these experimentally derived physiological traits onto a phylogeny and quantified the tendency for related species to be similar (phylogenetic signal). All traits related to Cd bioaccumulation and susceptibility exhibited statistically significant phylogenetic signal, although the signal strength varied among traits. Conventional and phylogenetically based regression models were compared, revealing great variability within orders but consistent, strong differences among insect families. Uptake and elimination rate constants were positively correlated among species, but only when effects of body size and phylogeny were incorporated in the analysis. Together, uptake and elimination rates predicted dramatic Cd bioaccumulation differences among species that agreed with field-based measurements. We discovered a potential tradeoff between the ability to eliminate Cd and the ability to detoxify it across species, particularly mayflies. The best-fit regression models were driven by phylogenetic parameters (especially differences among families) rather than functional traits, suggesting that it may eventually be possible to predict a taxon's physiological performance based on its phylogenetic position, provided adequate physiological information is available for close relatives. There appears to be great potential for evolutionary physiological approaches to augment our understanding of insect responses to environmental stressors in nature.comparative methods ͉ evolutionary physiology ͉ bioaccumulation ͉ phylogeny ͉ tradeoff W ith Ϸ6,500 species described to date in North America (1), aquatic insects are a diverse and ecologically important group (2), particularly in rivers and streams. For example, the orders Ephemeroptera, Plecoptera, and Trichoptera (EPT taxa) include 58 recognized families and Ϸ2,700 species (1). Among these many lineages, great diversity exists in morphology, life history characteristics, and physiology stemming from a long and complex evolutionary history. Although the origins of the Ephemeroptera are unknown (3), a general paradigm of the terrestrial ancestry of aquatic insects is widely accepted, with numerous invasions of freshwater habitats hypothesized throughout evolutionary history (4). Many of these invasions have entailed adaptive ''solutions'' that involve complex suites of traits that in combination determine the range of environmental conditions that a given taxon can tolerate.Some traits that arose in response to past environmental challenges may now render certain species relatively more susceptible to modern anthropogenic pollutants....

Although the differential responses of stream taxa to metal exposure have been exploited for bioassessment and monitoring, the mechanisms affecting these responses are not well understood. In this study, the subcellular partitioning of metals in operationally defined metal-sensitive and detoxified fractions were analyzed in five insect taxa. Samples were collected in two separate years along an extensive metal contamination gradient in the Clark Fork River (MT, USA) to determine if interspecific differences in the metal concentrations of metal-sensitive fractions and detoxified fractions were linked to the differences in distributions of taxa relative to the gradient. Most of the Cd, Cu, and Zn body burdens were internalized and potentially biologically active in all taxa, although all taxa appeared to detoxify metals (e.g., metal bound to cytosolic metal-binding proteins). Metal concentrations associated with metal-sensitive fractions were highest in the mayflies Epeorus albertae and Serratella tibialis, which were rare or absent from the most contaminated sites but occurred at less contaminated sites. Relatively low concentrations of Cu were common to the tolerant taxa Hydropsyche spp. and Baetis spp., which were widely distributed and dominant in the most contaminated sections of the river. This suggested that distributions of taxa along the contamination gradient were more closely related to the bioaccumulation of Cu than of other metals. Metal bioaccumulation did not appear to explain the spatial distribution of the caddisfly Arctopsyche grandis, considered to be a bioindicator of metal effects in the river. Thus, in this system the presence/ absence of most of these taxa from sites where metal exposure was elevated could be differentiated on the basis of differences in metal bioaccumulation.

Trace element bioaccumulation was studied in immature benthic insects from two contaminated river systems to develop these animals as bioindicators. In one river, Cu, Cd, Pb, and Zn were analysed in insects and in fine bed sediments over a 381-km reach downstream of a large copper mining complex. In the other river, As contamination from a gold mine was assessed in insects and bed sediments over a 40-km reach. All insect taxa collected in contaminated river reaches had elevated whole-body trace element concentrations. However, direct comparisons of contamination using a single, common species among stations were limited because few species were distributed throughout the study reaches. Comparisons of contamination at taxomic levels higher than species were complicated by element-specific differences in bioaccumulation among taxa. These differences appeared to be governed by biological and hydrogeochemical factors. The variation in element concentrations among species of the caddisfly Hydropsyche was slightly greater than within individual species. If this genus is representative of others, comparisons of contamination within genera may be a practical alternative for biomonitoring studies when single species are not available.

Consumption of periphyton is a potentially important route of metal exposure to benthic invertebrate grazers. The present study examined the bioaccumulation kinetics of dissolved and dietary Cd and Cu in five species of mayflies (class Insecta). Artificial stream water and benthic diatoms were separately labeled with enriched stable metal isotopes to determine physiological rate constants used by a biokinetic bioaccumulation model. The model was employed to simulate the effects of metal partitioning between water and food, expressed as the bioconcentration factor (BCF), as well as ingestion rate (IR) and metal assimilation efficiency of food (AE), on the relative importance of water and food to metal bioaccumulation. For all test species, the contribution of dietary uptake of Cd and Cu increased with BCF. For a given BCF, the contribution of food to the body burden increased with k(uf) , the metal uptake rate constant from food that combined variation in IR and AE. To explore the relative importance of water and diet exposure routes under field conditions, we used estimated site-specific aqueous free-ion concentrations to model Cd and Cu accumulation from aqueous exposure, exclusively. The predicted concentrations accounted for less than 5% of the observed concentrations, implying that most bioaccumulated metal was acquired from food. At least for the taxa considered in this study, we conclude that consumption of metal-contaminated periphyton can result in elevated metal body burdens and potentially increase the risk of metal toxicity.