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. The rapidly intensifying process of ocean acidification (OA) due toanthropogenic CO2 is not only depleting carbonate ions necessaryfor calcification but also causing acidosis and disrupting internal pHhomeostasis in several marine organisms. These negative consequences of OA onmarine calcifiers, i.e. oyster species, have been very well documented inrecent studies; however, the consequences of reduced or impairedcalcification on the end-product, shells or skeletons, still remain one ofthe major research gaps. Shells produced by marine organisms under OA areexpected to show signs of dissolution, disorganized microstructure andreduced mechanical properties. To bridge this knowledge gap and to test theabove hypothesis, we investigated the effect of OA on juvenile shells of thecommercially important oyster species, Magallana angulata,at ecologically and climatically relevant OA levels (using pH 8.1, 7.8, 7.5,7.2). In lower pH conditions, a drop of shell hardness and stiffness wasrevealed by nanoindentation tests, while an evident porous internalmicrostructure was detected by scanning electron microscopy. Crystallographicorientation, on the other hand, showed no significant difference withdecreasing pH using electron back-scattered diffraction (EBSD). These resultsindicate the porous internal microstructure may be the cause of the reductionin shell hardness and stiffness. The overall decrease of shell density observedfrom micro-computed tomography analysis indicates the porous internalmicrostructure may run through the shell, thus inevitably limiting theeffectiveness of the shell's defensive function. This study shows the potentialdeterioration of oyster shells induced by OA, especially in their early lifestage. This knowledge is critical to estimate the survival and production ofedible oysters in the future ocean.

Ocean acidification reduces hardness and stiffness of the Portuguese oyster shell with impaired microstructure: a hierarchical analysis