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Using a selection of molecular biomarkers, we evaluated responses in freshwater mussels ( Anodonta anatina ) exposed to effluent from an industrial wastewater treatment facility. The aims of this work were to (1) assess biomarkers of general toxicity under sublethal exposure to an anthropogenic mixture of chemicals, represented by an arbitrary effluent, and (2) evaluate the potential of  A. anatina  as a bioindicator of pollution. Adult mussels ( n =  in total 32; 24 males and 8 females) were exposed (96 h) in the laboratory to a fixed dilution of effluent or to a control treatment of standardized freshwater. Metal concentrations were in general higher in the effluent, by an order of magnitude or more, compared to the control. Toxic unit estimates were used as proxies of chemical stress, and Cu, Ni, and Zn were identified as potential major contributors (Cu&gt; Ni &gt; Zn). Six transcriptional ( cat ,  gst ,  hsp70 ,  hsp90 ,  mt ,  sod ) and two biochemical (AChE, GST) biomarkers were analyzed in two tissues, gills, and digestive glands. Out of the 16 responses (eight biomarkers × two tissues), 14 effect sizes were small (within ± 28 % of control) and differences non-significant ( p  &gt; 0.05). Results did however show that (1) AChE activity increased by 40% in gills of exposed mussels compared to control, (2)  hsp90  expression was 100% higher in exposed female gills compared to control, and (3) three marker signals (AChE in both tissues, and  hsp70  in gills) differed between sexes, independent of treatment. Results highlight a need for further investigation of molecular biomarker variability and robustness in  A. anatina .

Molecular biomarker responses in the freshwater mussel Anodonta anatina exposed to an industrial wastewater effluent