Beschreibung: In coastal temperate regions such as the Baltic Sea, calcifying bivalves dominate benthic communities playing a vital ecological role in maintaining biodiversity and nutrient recycling. At low salinities, bivalves exhibit reduced growth and calcification rates which is thought to result from physiological constraints associated with osmotic stress. Calcification demands a considerable amount of energy in calcifying molluscs and estuarine habitats provide sub-optimal conditions for calcification due to low concentrations of calcification substrates and large variations in carbonate chemistry. Therefore, we hypothesize that slow growth rates in estuarine bivalves result from increased costs of calcification, rather than costs associated with osmotic stress. To investigate this, we estimated the cost of calcification for the first time in benthic bivalve life stages and the relative energy allocation to calcification in three Mytilus populations along the Baltic salinity gradient. Our results indicate that calcification rates are significantly reduced only in 6 psu populations compared to 11 and 16 psu populations, coinciding with ca. 2–3-fold higher calcification costs at low salinity and temperature. This suggests that reduced growth of Baltic Mytilus at low salinities results from increased calcification costs rather than osmotic stress related costs. We also reveal that shell growth (both calcification and shell organic production) demands 31–60% of available assimilated energy from food, which is significantly higher than previous estimates. Energetically expensive calcification represents a major constraint on growth of mytilids in the estuarine and coastal seas where warming, acidification and desalination are predicted over the next century.

Beschreibung: Background: The green crab Carcinus maenas is known for its high acclimation potential to varying environmental abiotic conditions. A high ability for ion and acid-base regulation is mainly based on an efficient regulation apparatus located in gill epithelia. However, at present it is neither known which ion transport proteins play a key role in the acid-base compensation response nor how gill epithelia respond to elevated seawater pCO2 as predicted for the future. In order to promote our understanding of the responses of green crab acid-base regulatory epithelia to high pCO2, Baltic Sea green crabs were exposed to a pCO2 of 400 Pa. Gills were screened for differentially expressed gene transcripts using a 4,462-feature microarray and quantitative real-time PCR. Results: Crabs responded mainly through fine scale adjustment of gene expression to elevated pCO2. However, 2% of all investigated transcripts were significantly regulated 1.3 to 2.2-fold upon one-week exposure to CO2 stress. Most of the genes known to code for proteins involved in osmo- and acid-base regulation, as well as cellular stress response, were were not impacted by elevated pCO2. However, after one week of exposure, significant changes were detected in a calcium-activated chloride channel, a hyperpolarization activated nucleotide-gated potassium channel, a tetraspanin, and an integrin. Furthermore, a putative syntaxin-binding protein, a protein of the transmembrane 9 superfamily, and a Cl-/HCO3 - exchanger of the SLC 4 family were differentially regulated. These genes were also affected in a previously published hypoosmotic acclimation response study. Conclusions: The moderate, but specific response of C. maenas gill gene expression indicates that (1) seawater acidification does not act as a strong stressor on the cellular level in gill epithelia; (2) the response to hypercapnia is to some degree comparable to a hypoosmotic acclimation response; (3) the specialization of each of the posterior gill arches might go beyond what has been demonstrated up to date; and (4) a re-configuration of gill epithelia might occur in response to hypercapnia.

Beschreibung: Background: Biomineralization by molluscs involves regulated deposition of calcium carbonate crystals within a protein framework to produce complex biocomposite structures. Effective biomineralization is a key trait for aquaculture, and animal resilience under future climate change. While many enzymes and structural proteins have been identified from the shell and in mantle tissue, understanding biomieralization is impeded by a lack of fundamental knowledge of the genes and pathways involved. In adult bivalves, shells are secreted by the mantle tissue during growth, maintenance and repair, with the repair process, in particular, amenable to experimental dissection at the transcriptomic level in individual animals. Results: Gene expression dynamics were explored in the adult blue mussel, Mytilus edulis, during experimentally induced shell repair, using the two valves of each animal as a matched treatment-control pair. Gene expression was assessed using high-resolution RNA-Seq against a de novo assembled database of functionally annotated transcripts. A large number of differentially expressed transcripts were identified in the repair process. Analysis focused on genes encoding proteins and domains identified in shell biology, using a new database of proteins and domains previously implicated in biomineralization in mussels and other molluscs. The genes implicated in repair included many otherwise novel transcripts that encoded proteins with domains found in other shell matrix proteins, as well as genes previously associated with primary shell formation in larvae. Genes with roles in intracellular signalling and maintenance of membrane resting potential were among the loci implicated in the repair process. While haemocytes have been proposed to be actively involved in repair, no evidence was found for this in the M. edulis data. Conclusions: The shell repair experimental model and a newly developed shell protein domain database efficiently identified transcripts involved in M. edulis shell production. In particular, the matched pair analysis allowed factoring out of much of the inherent high level of variability between individual mussels. This snapshot of the damage repair process identified a large number of genes putatively involved in biomineralization from initial signalling, through calcium mobilization to shell construction, providing many novel transcripts for future in-depth functional analyses

Beschreibung: Baltic blue mussels can colonise and dominate habitats with far lower salinity (〈 10 psu) than other Mytilus congeners. Pervasive gene flow was observed between Western Baltic Mytilus edulis living at high salinity conditions and Eastern Baltic M. trossulus living at lower salinites, with highest admixture proportions within a genetic transition zone located at intermediate salinities (Darss Sill area). Yet, we do not understand the impacts of low salinity on larval performance, and how salinity may act as an early selective pressure during passive larval drift across salinity gradients. This study tested whether larvae originating from two different populations along the natural salinity cline in the Baltic Sea have highest fitness at their native salinities. Our results suggest that Eastern Baltic M. trossulus (Usedom, 7 psu) and Western Baltic M. edulis (Kiel, 16 psu) larvae display better performance (fitness components: growth, mortality, settlement success) when reared at their respective native salinities. This suggests that these populations are adapted to their local environment. Additionally, species diagnostic markers were used for genetic analyses of transition zone (Ahrenshoop, 11 psu) mussel larvae exposed to low salinity. This revealed that low salinity selection resulted in a shift towards allele frequencies more typical for Eastern Baltic M. trossulus. Thus, salinity acts as a selective pressure during the pre-settlement phase and can shape the genetic composition of Baltic mussel populations driving local adaptation to low salinity. Future climate change driven desalination, therefore, has the potential to shift the Baltic Sea hybrid gradient westward with consequences for benthic ecosystem structure.

Beschreibung: 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.

Beschreibung: Salinity is a major environmental factor shaping the distribution and abundance of marine organisms. Climate change is predicted to alter salinity in many coastal regions due to sea level rise, evaporation, and changes in freshwater input. This exerts significant physiological stress on coastal invertebrates whose body fluid osmolality follows that of seawater ('osmoconformers'). In this study, we conducted a systematic review and meta-analysis of osmolytes (both organic and inorganic) utilized by osmoconforming marine invertebrates during a 〉 14-day acclimation to reduced salinity. Of the 2,389 studies screened, a total of 56 fulfilled the search criteria. Thirty-eight studies reported tissue osmolyte. Following acclimation to reduced salinity, tissue concentrations of six organic compounds and sodium were consistently reduced across phyla. This suggests that intracellular inorganic ions are not only utilized as a rapid response system during acute exposure to low salinity stress but also, in concert with reductions in organic osmolyte concentrations, during longer-term acclimation. Our systematic review demonstrates that only a few studies (n = 13) have quantified salinity-induced long-term changes in intracellular ion concentrations. In addition, no study has compiled a complete intracellular osmolyte budget. Alanine, betaine, glycine, and taurine are the major organic osmolytes that are universally employed across five phyla. The characterization of organic osmolytes was heavily weighted towards free amino acids (FAAs) and derivatives-neglecting methylamines and methylsulfonium compounds, which can be as important as FAAs in modulating intracellular osmolality. As a consequence, we suggest best-practice guidelines to streamline experimental designs and protocols in osmoregulation research in order to better understand the conserved mechanisms that define the limits of salinity acclimation in marine invertebrates. To our best knowledge, this is the first systematic review and meta-analysis on osmolyte concentrations in osmoconformers acclimated to low salinity. It creates a valuable baseline for future research and reveals large research gaps. Our meta-analysis suggests that there are common osmolyte actors employed across phyla but no uniform concept since osmolyte pool composition and proportions were taxon-specific. In light of future salinity changes and their potential consequences, it becomes more important to understand salinity tolerance capacities and limits.