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We studied how copper toxicity in the red flour beetle, Tribolium castaneum changed as a result of infection by the entomopathogenic nematode Steinernema feltiae. Measured traits were: respiration, growth and survival, as well as the concentrations of copper within beetle tissues and in its diet. By comparing F1 and F5 generation we were able to answer how long-term metal exposure changed the responses to both copper and the parasite. The beetles did accumulate copper; however, the results indicated that copper concentrations in beetle tissues were affected by nematode infection, the sex of the experimental animals, and the number of generations of exposure. Five generations of exposure to copper resulted in the highest dry body mass of infected beetles of both sexes; additionally, this group also had the lowest copper concentrations in their tissues. The only factor that had a significant effect on respiration was infection by nematodes: infected beetles of both sexes in both generational groups had significantly decreased respiration rates. Survival was lowest in nematode-infected animals of both sexes from both generations, regardless of exposure to copper. Our results confirm that an organism's response to metal pollution is dependent on its health status and sex. We also found that the history of exposure to metal was equally important-we found enhanced resistance to copper intoxication after only five generations of exposure.

Temperature is a key environmental factor affecting almost all aspects of life history in ectotherms. Theory predicts that they grow faster, reach smaller sizes and produce smaller offspring when temperature increases. In addition, temperature changes, through their effects on metabolism, may also influence the expression of alternative reproductive phenotypes (ARPs) in ectotherms. Although many studies have investigated the phenotypic plasticity of life history traits in relation to temperature change, little is known about how those traits and phenotypic plasticity may evolve together. In our study we subjected bulb mites (non-model, soil organisms that normally experience rather stable thermal conditions) to experimental evolution in two temperature treatments: control (24°C) and elevated (28°C). After 18 generations, we measured adult body size, egg size and development time of both treatments at control as well as at elevated temperatures (test temperatures). Thus, we were able to detect genetic changes (the effects of selection temperature) and environmental effects (the effects of test temperature). We also observed the ARP expression throughout the experimental evolution. Our results revealed quite complex patterns of life history in traits response to temperature. Mites developed faster and reached smaller sizes at increased temperature, but evolutionary responses to increased temperature were not always parallel to the observed phenotypic plasticity. Additionally, despite smaller body sizes, females laid larger eggs at higher temperature. This effect was more pronounced in animals evolving at elevated temperature. Evolution at increased temperature also affected ARP expression, with the proportion of armored fighters decreasing from generation to generation. We propose that this could be the consequence of temperature sensitivity of the cost-to-benefits ratio of expressing ARPs.

Ecotoxicological effect studies often expose test organisms under optimal environmental conditions. However, organisms in their natural settings rarely experience optimal conditions. On the contrary, during most of their lifetime they are forced to cope with sub-optimal conditions and occasionally with severe environmental stress. Interactions between the effects of a natural stressor and a toxicant can sometimes result in greater effects than expected from either of the stress types alone. The aim of the present review is to provide a synthesis of existing knowledge on the interactions between effects of "natural" and chemical (anthropogenic) stressors. More than 150 studies were evaluated covering stressors including heat, cold, desiccation, oxygen depletion, pathogens and immunomodulatory factors combined with a variety of environmental pollutants. This evaluation revealed that synergistic interactions between the effects of various natural stressors and toxicants are not uncommon phenomena. Thus, synergistic interactions were reported in more than 50% of the available studies on these interactions. Antagonistic interactions were also detected, but in fewer cases. Interestingly, about 70% of the tested chemicals were found to compromise the immune system of humans as judged from studies on human cell lines. The challenge for future studies will therefore be to include aspects of combined stressors in effect and risk assessment of chemicals in the environment., (Copyright 2009 Elsevier B.V. All rights reserved.)

The paper addresses problems arising from effects of natural environmental factors on toxicity of pollutants to organisms. Most studies on interactions between toxicants and natural factors, including those completed in the EU project NoMiracle (Novel Methods for Integrated Risk Assessment of Cumulative Stressors in Europe) described herein, showed that effects of toxic chemicals on organisms can differ vastly depending purely on external conditions. We compiled data from 61 studies on effects of temperature, moisture and dissolved oxygen on toxicity of a range of chemicals representing pesticides, polycyclic aromatic hydrocarbons, plant protection products of bacterial origin and trace metals. In 62.3% cases significant interactions (p< or =0.05 or less) between natural factors and chemicals were found, reaching 100% for the effect of dissolved oxygen on toxicity of waterborne chemicals. The meta-analysis of the 61 studies showed that the null hypothesis assuming no interactions between toxic chemicals and natural environmental factors should be rejected at p=2.7 x 10(-82) (truncated product method probability). In a few cases of more complex experimental designs, also second-order interactions were found, indicating that natural factors can modify interactions among chemicals. Such data emphasize the necessity of including information on natural factors and their variation in time and across geographic regions in ecological risk assessment. This can be done only if appropriate ecotoxicological test designs are used, in which test organisms are exposed to toxicants at a range of environmental conditions. We advocate designing such tests for the second-tier ecological risk assessment procedures., (Copyright 2010 Elsevier B.V. All rights reserved.)

Terrestrial organisms in the field often are exposed to a combination of stress factors of various origins, but little is known about interactions between different types of stressors. In the present study, we demonstrate the results of a study on interactions between Ni, chlorpyrifos (CPF), and temperature in the ground beetle, Pterostichus oblongopunctatus. The results revealed that all factors, and their interactions, influenced life-cycle parameters of the beetles (survival and reproduction). Significant three-factor interactions were found for effects on beetle survival, indicating that the combined negative effect of Ni and CPF was temperature dependent. In addition, significant effects of body mass were found: The survival of beetles treated with CPF and the reproduction of beetles exposed to Ni were positively correlated with body mass. All studied endpoints were affected by temperature. The results indicate that understanding interactions between temperature and toxicants, as well as among chemicals themselves, is essential for proper ecological risk assessment.

To determine the effect of nematode infection on the response of snails to selected toxins, we infected Helix aspersa with 0-, 0.25-, 1-, or 4-fold the recommended field dose of a commercial nematode application for agricultural use. In the first experiment, the snails also were exposed to cadmium via food and soil at concentrations of 0, 30, 60, 120, or 240 mg/kg in a full-factorial design. In the second experiment, snails were infected with nematodes and also fed either Bt (expressing Bacillus thuringiensis toxin) maize or non-Bt maize. The snails were weighed at the beginning and end (after four weeks) of the experiments, and mortality was checked daily. Neither exposure of snails to nematodes nor exposure of snails to cadmium or Bt toxin affected the survival rates of snails. The number of dead snails was highest for combinations of nematode treatments with cadmium concentrations of 120 and 240 mg/kg. In both experiments (Bt and cadmium), the growth rate decreased with increasing nematode dose. The Bt maize was not harmful to the snails in the absence of nematodes, but infected snails grew faster when fed non-Bt maize. The growth rate of snails exposed to cadmium decreased with exposure to increasing Cd concentrations and differed significantly between the no-nematode treatment and the treatments with nematode doses of one- and fourfold the recommended field dose. Snails treated with the highest dose of nematodes accumulated the highest cadmium concentrations.