A window into the effect of ocean acidification on molluscan larval shell development using a quantitative approach

ABSTRACT Increasing atmospheric CO2 levels have led to decreased pH and calcium carbonate saturation (Ω) of seawater, a process referred to as ocean acidification. Ocean acidification is expected to reduce biomineralization by marine calcifiers, such as molluscs, and many studies have reported serious effects on molluscan shell development. However, it has not previously been possible to quantitatively compare these effects on tiny structures, such as larval shells, among and within species. We applied the measurement technique of micro-focus X-ray computed tomography (MXCT) to larval shells of the limpet Nipponacmea fuscoviridis to quantitatively trace the process of shell growth (shell thickness and shell density). Shell thickness and density significantly decreased in seawater with low Ω levels. Scanning electron microscopy (SEM) revealed that the surface structure of the shell in larvae cultured under low Ω was disturbed. Gene expression analysis showed that the development of shell-forming regions under low Ω was significantly reduced. MXCT analysis can quantify mineralization in tiny larval shells; in combination with other methods such as SEM and gene expression analysis, it can provide a novel perspective in the assessment of the impact and resilience of marine calcifiers to changes in the marine environment.