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Abstract: Humic substances comprise the majority of natural organic matter (NOM) on Earth, including dissolved organic matter in freshwater systems. Recent studies show that these substances directly interact with aquatic organisms as chemical stressors. The aim of the present study was to investigate the mode of action of dissolved NOM on the freshwater amphipods Gammarus lacustris Sars and Gammarus tigrinus (Sexton), and in particular, to determine if NOM induces or promotes internal oxidative stress. NOM was isolated by reverse osmosis from a brown-water lake in Brandenburg State, Germany. Oxidative stress markers, such as lipid peroxidation, cell internal hydrogen peroxide concentration, as well as peroxidase, catalase and glutathione S-transferase activities, were quantified. Exposure of both amphipod species to NOM caused a significant increase in lipid peroxidation, hydrogen peroxide concentration, catalase, peroxidase and glutathione S-transferase activities. Both species showed a two-stage antioxidant response: the first stage allowed the organisms to effectively eliminate ROS and to protect cells from damage, whereas the second stage leads to H2O2 accumulation in combination with destruction of lipid structures in the cells and, finally, functional damage or even death of the organism. [Copyright &y& Elsevier]

Natural organic matter (NOM) isolated from the eutrophic Sanctuary Pond (Point Pelee National Park, Canada) has an adverse impact on amphipod species (Gammarus tigrinus and Chaetogammarus ischnus from Lake Mu¨ggelsee, Germany, and Eulimnogammarus cyaneus, from Lake Baikal, Russia). Increases in amphipod mortality, changes in peroxidase activity and increases of heat shock protein (hsp70) expression were observed upon exposure to NOM. The highest resistance to the adverse impact of NOM was observed with the endemic Baikalian amphipod E. cyaneus. However, the mechanisms behind this finding remains obscure. If differences in the sensitivity of the hsp70 antibody may be excluded, different modes of action may be postulated: because the adverse impact of NOM may be caused by reactive oxygen species (ROS) and the NOM itself, the observed differences may be due to the action of ROS alone (with E. cyaneus) and a combination of both adverse modes of action (European species). [Copyright &y& Elsevier]

The ancient Lake Baikal is the largest source of liquid freshwater on Earth and home to a unique fauna. Several hundred mostly cold-adapted endemic amphipod species inhabit Baikal, an ecosystem that is already being influenced by global change. In this study, we characterized the core proteome and heat stress-induced changes in a temperature-tolerant endemic amphipod, Eulimnogammarus cyaneus , using a proteogenomic approach (PRIDE dataset PXD013237) to unravel the molecular mechanisms of the observed adverse effects. As males were previously found to be much more tolerant to thermal stress, we placed special emphasis on differences between the sexes. For both sexes, we observed adaption of energy metabolism, cytoskeleton, lipid, and carbohydrate metabolism upon heat stress. In contrast, significant differences were determined in the molecular chaperone response. Females from the control conditions possessed significantly higher levels of heat shock proteins (HSP70, HSPb1, Hsc70-3), which, in contrast to males, were not further increased in response to heat stress. The inability of females to further increase heat shock protein synthesis in response to temperature stress may be due to sex-specific processes, such as egg production, requiring a large proportion of the available energy. As ovigerous females synthesize generally higher amounts of protein, they also need higher levels of molecular chaperones for the folding of these new proteins. Thus, the higher sensitivity of females to heat shock may be due to the lack of molecular chaperone molecules to counteract the heat-induced protein denaturation. Unlabelled Image • Endemic amphipods are extremely important in benthic communities of Lake Baikal. • We studied the proteomes of males and females of Eulimnogammarus cyaneus. • High-throughput mass spectrometry allowed us to find almost 1000 protein groups. • Females possess higher levels of heat shock proteins in control conditions. • Males activate heat shock proteins in heat stress conditions, while females do not. [ABSTRACT FROM AUTHOR]