Inorganic ion and metabolite concentrations in tissue of marine invertebrates exposed to low salinity in the laboratory

Laboratory experiments were conducted in the climate chambers at GEOMAR Helmholtz Centre for Ocean Research Kiel in the time between March and November 2018. Experiments were designed to study the effect of long-term (1 month) exposure to low salinity in osmoconforming invertebrates. The study organisms (Asterias rubens, Mytilus edulis, Littorina littorea, Diadumene lineata, Strongylocentrotus droebachiensis and Psammechinus milliaris) were collected in Kiel Fjord, Eckernförder Bight or the Kattegat from spring to autumn 2018. Organisms were acclimated to climate chamber conditions for 1 week (under habitat salinity, 14˚C, constant aeration) and then subjected to salinity acclimation for 1-2 weeks until the final salinity treatment level was reached. Then different salinity treatments were maintained for 4 weeks. Water physiochemistry (temperature, salinity, pH, nitrite, nitrate, phosphate) was recorded frequently. After the experiment, samples were taken from tissues to measure total osmolality (mosmol/kg) with an osmomat, and inorganic ions (mmol/kg or µmol/g wet mass). Anions were measured with a novel protocol via ion chromatography, cations were measured via flame photometry. Organic osmolytes were measured via 1H-NMR.