Description: Low-salinity stress can severely affect the fitness of marine organisms. As desalination has been predicted for many coastal areas with ongoing climate change, it is crucial to gain more insight in mechanisms that constrain salinity acclimation ability. Low-salinity induced depletion of the organic osmolyte pool has been suggested to set a critical boundary in osmoconforming marine invertebrates. Whether inorganic ions also play a persistent role during low-salinity acclimation processes is currently inconclusive. We investigated the salinity tolerance of six marine invertebrate species following a four-week acclimation period around their low-salinity tolerance threshold. To obtain complete osmolyte budgets, we quantified organic and inorganic osmolytes and determined fitness proxies. Our experiments corroborated the importance of the organic osmolyte pool during low-salinity acclimation. Methylamines constituted a large portion of the organic osmolyte pool in molluscs, whereas echinoderms exclusively utilized free amino acids. Inorganic osmolytes were involved in long-term cellular osmoregulation in most species, thus are not just modulated with acute salinity stress. The organic osmolyte pool was not depleted at low salinities, whilst fitness was severely impacted. Instead, organic and inorganic osmolytes often stabilized at low-salinity. These findings suggest that low-salinity acclimation capacity cannot be simply predicted from organic osmolyte pool size. Rather, multiple parameters (i.e. osmolyte pools, net growth, water content and survival) are necessary to establish critical salinity ranges. However, a quantitative knowledge of cellular osmolyte systems is key to understand the evolution of euryhalinity and to characterize targets of selection during rapid adaptation to ongoing desalination.

Description: 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.

Description: Low-salinity stress can severely affect the fitness of marine organisms. As desalination has been predicted for many coastal areas with ongoing climate change, it is crucial to gain more insight in mechanisms that constrain salinity acclimation ability. Low-salinity induced depletion of the organic osmolyte pool has been suggested to set a critical boundary in osmoconforming marine invertebrates. Whether inorganic ions also play a persistent role during low-salinity acclimation processes is currently inconclusive. We investigated the salinity tolerance of six marine invertebrate species following a four-week acclimation period around their low-salinity tolerance threshold. The species investigated were Asterias rubens, Mytilus edulis, Littorina littorea, Diadumene lineata, Strongylocentrotus droebachiensis and Psammechinus milliaris. To obtain complete osmolyte budgets of seawater, body fluids and tissues we quantified total osmolality (via osmometer), organic osmolytes (methylamine and free amino acids) via 1H-NMR spectroscopy and inorganic osmolytes (anions and cations) via flame photometry and a novel protocol using ion-chromatography. We further determined the fitness proxies survival, growth and tissue water content. Our data show the importance of the organic and inorganic osmolyte pool during low-salinity acclimation. It also shows the importance of specific compounds in some species. This data can be used in future osmolyte and salinity tolerance research. This type of data is essential to establish reliable physiological limits of species in order to estimate consequences of future salinity changes with ongoing climate change. It can be used to assess the salinity tolerance capacity and to obtain a better understanding of the basic mechanisms that are utilized in a wide range of species. The established cellular inorganic and organic osmolyte profiles can build a foundation for applied cellular physiological research, for example for designing suitable buffers for in vitro assays as these buffers need to incorporate complex organic and inorganic osmolyte changes. Knowledge about cellular and whole-organism biochemistry and physiology is absolutely crucial for characterizing the functions of genes that are under selection by climate change stressors. A quantitative knowledge of cellular osmolyte systems is key to understand the evolution of euryhalinity and to characterize targets of selection during rapid adaptation to ongoing desalination.

Description: 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. Throughout the experiment survival was recorded. Before and after the experiment organism weight and, where feasible, size was measured. Weight data was also used to calculate tissue water content. This dataset comprises the physiological fitness parameters for each species at the respective salinity treatment. Given are data for survival, growth and tissue water content.

Description: 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 seawater and body fluids to measure total osmolality (mosmol/kg) with an osmomat and inorganic ions (mmol/l). No body fluid samples were taken from Diadumene lineata as organisms were too small and volumes too low. Anions were measured with a novel protocol via ion chromatography, cations were measured via flame photometry.

Description: Low-salinity stress can severely affect the fitness of marine organisms. As desalination has been predicted for many coastal areas with ongoing climate change, it is crucial to gain more insight in mechanisms that constrain salinity acclimation ability. Low-salinity induced depletion of the organic osmolyte pool has been suggested to set a critical boundary in osmoconforming marine invertebrates. Whether inorganic ions also play a persistent role during low-salinity acclimation processes is currently inconclusive. We investigated the salinity tolerance of six marine invertebrate species following a four-week acclimation period around their low-salinity tolerance threshold. To obtain complete osmolyte budgets, we quantified organic and inorganic osmolytes and determined fitness proxies. Our experiments corroborated the importance of the organic osmolyte pool during low-salinity acclimation. Methylamines constituted a large portion of the organic osmolyte pool in molluscs, whereas echinoderms exclusively utilized free amino acids. Inorganic osmolytes were involved in long-term cellular osmoregulation in most species, thus are not just modulated with acute salinity stress. The organic osmolyte pool was not depleted at low salinities, whilst fitness was severely impacted. Instead, organic and inorganic osmolytes often stabilized at low-salinity. These findings suggest that low- salinity acclimation capacity cannot be simply predicted from organic osmolyte pool size. Rather, multiple parameters (i.e. osmolyte pools, net growth, water content and survival) are necessary to establish critical salinity ranges. However, a quantitative knowledge of cellular osmolyte systems is key to understand the evolution of euryhalinity and to characterize targets of selection during rapid adaptation to ongoing desalination.