Evaluation of methods for assessing physiological biomarkers of stress in freshwater mussels
Freshwater mussel populations are highly susceptible to environmental alterations because of their diminished numbers and primarily sessile behaviors; nonlethal biomonitoring programs are needed to evaluate the health of populations prior to mass mortality events. Our objectives were to determine ( i ) which biochemical parameters in freshwater mussel hemolymph could be consistently quantified, ( ii ) how hemolymph parameters and tissue glycogen respond to a thermal stress gradient (25, 30, and 35 °C), and ( iii ) the effects of tissue and hemolymph extraction on long-term growth and survival of smaller- and larger-bodied mussel species. Glucose exhibited elevated expression in both species with increasing water temperature. Two transaminase enzymes had elevated expression in the 30 °C treatment. The effects of hemolymph extraction and tissue biopsies were evaluated with a large-bodied species, Elliptio crassidens , and a smaller species, Villosa vibex . Individuals were monitored for 820 to 945 days after one of four treatments: hemolymph extraction, tissue biopsy, tissue and hemolymph extraction, and control. Hemolymph extraction and tissue biopsy adversely affected survival of V. vibex , suggesting that these extraction methods may add some risk of reduced survival to smaller-bodied species. Survival of E. crassidens was not impaired by any of the treatments, supporting the use of these techniques in nonlethal biomonitoring programs for larger-bodied mussel species.