Chamid bivalves are marine organisms that live in high-energy environments and are cemented to hard substrates. To avoid shell damage, the organisms form thick, densely ornamented shells. Shell material consists of aragonite, and the ornamentation may be either aragonitic or calcitic. The latter can be developed as scaly spines, rows of blades, or comarginal, radial arched lamellae. We investigated biological crystal morphology and mode of assembly of Chama arcana and Chama gryphoides shells. Structural characteristics were obtained from electron backscatter diffraction (EBSD) measurements, complemented with laser confocal and BSE imaging. We found a wide range of crystal morphologies and sizes, ranging from irregularly shaped calcite and/or aragonite prisms to tiny and thin aragonite laths. We observed four different modes of crystal assembly patterns: 1. strongly interlocked dendritic calcite units forming the ornamentation blades; 2. aragonite laths arranged to lamellae forming the outer shell layer, the layer adjacent to the calcite; 3. aragonite laths arranged into blocks comprising inner shell layers or aragonitic ornamentations; and 4. shell portions consisting of aragonite prisms, structured in size and crystal orientation, at muscle attachment sites. These four different types of crystal arrangements were observed for the shells of the investigated chamid species; however, they had slightly different strengths of structuring and slight variations in crystal organisation. Additionally, we observed unique microstructural features in Chama shells: We report ornamentation crystals resembling idiomorphic calcite and novel, twinned entities found at the changeover between the aragonitic layers. We highlight and discuss these differences and anomalies in this contribution.

Recent advancements in electron backscatter diffraction (EBSD) data evaluation enable the determination of misorientation between crystals below 0.1°, while with conventional EBSD data evaluation, the smallest misorientation precision between crystals scatters between 0.5°-1°. Sea urchin tests and spines are lightweight biomaterials with a serrated microstructure comprising interlinked calcite crystals. We investigated the microstructure and crystallographic texture of Cidaris cidaris and Paracentrotus lividus test and spine calcite with advanced EBSD measurement and data evaluation. In particular, we re-evaluated the widely accepted single-crystallinity of sea urchin calcite. We found that the test and the spines comprise calcite crystals with different fabrics and a significant variation in crystal co-orientation strength. Even the highly co-oriented calcite of C. cidaris and P. lividus is not perfectly single-crystalline. We found test and spine portions that feature significant internal misorientations (1-3°). Test c-axis orientation in C. cidaris is tangential to the outer test surface, while in the spines, it is parallel to the morphological axis of the spine. Primary and secondary spines feature a bimodal crystal texture comprising co-oriented calcite surrounded by a cortex of misoriented crystals. Crystal misorientation in the spine cortex seems to result mainly from competitive growth determinants. Deciphering the degree of crystallinity and mode of crystal organisation of biological hard tissues is vital for understanding their exceptional control of structure, material architecture and material properties. STATEMENT OF SIGNIFICANCE: Echinoids form lightweight biomineralised skeletal elements with outstanding material properties and a complex microstructure formed of interlinked calcite crystals. The degree of crystallinity and the crystallographic organisation of the calcitic tests and spines are still under debate. We investigate and discuss the crystallinity, microstructure, and texture of Cidaris cidaris and Paracentrotus lividus test and spine crystals. Unprecedented and not yet used for biomineralised carbonate tissues, we apply electron backscatter diffraction pattern matching data evaluation, enabling detection of misorientation precision below 0.1°, relative to 0.5°-1° misorientation precision obtained from conventional EBSD data evaluation. We demonstrate that sea urchin test plates and spines are not single crystals. They feature internal small-angle misorientations and poorly co-oriented, polycrystalline regions with intricate microstructures.

Scaphopod molluscs are marine, infaunal, cosmopolitan animals. They encase their soft tissue with an aragonitic tusk-shaped shell that has at its two ends an orifice, the aperture and the apex. We investigated the shell of the dentaliid species Fissidentalium metivieri and Antalis weinkauffi and characterized shell crystal organization with electron backscatter diffraction (EBSD) and laser confocal and scanning electron microscopies. Based on crystal size, morphology, organization and growth-line spacing, we distinguish five different crystal arrangement motifs in the investigated shells. We find two slightly different microstructures for the shell proper, two microstructures for the attachment of muscles to the shell and one microstructure for a secondary hard tissue growth product, secreted at the apical orifice. Crystal organization motifs are crossed-lamellar, dendritic and prismatic. Crystal textures are crossed-lamellar, axial-like and crossed-lamellar-like. Crystal organization with a crossed arrangement is utilized for shell formation by representatives of many Ca-carbonate shell-secreting mollusc classes/subclasses: Scaphopoda, Gastropoda, Bivalvia, Polyplacophora (crossed-lamellar), and Patellogastropoda (crossed-foliated). Based on structural–crystallographic attributes, we highlight a basic shell structure that is broadly similar for the species of these mollusc classes/subclasses. However, this basic crystal arrangement motif is significantly modulated by the specific crystal organization that is inherent for a particular mollusc class/subclass.

The movement of valves of bivalved invertebrates is enabled through the action of muscles and the interplay between the muscles and the hinge ligament. The muscles that move the valves attach to their internal surface. To promote the structural integrity at the mechanically mismatched interfaces, a specific crystal microstructure and texture are present at the muscle attachment sites. These are different from the crystal microstructure and texture of the rest of the valves. We present here for modern two- and three-layered brachiopod shells (Magellania venosa, Liothyrella neozelanica and Gryphus vitreus) the mode of crystal organisation at sites of adductor and diductor muscle attachments (i) relative to the microstructure and texture that forms the other sections of the valves and (ii) relative to crystal organisation of muscle attachment sites of bivalved invertebrates of other phyla, namely, species of the class Bivalvia. We discuss similarities/differences in Ca-carbonate phase, microstructure and texture between rhynchonellate brachiopods and bivalves, and discuss whether the Ca-carbonate crystal organisation of muscle attachment sites is convergent for bivalved marine organisms. We show significant differences in muscle attachment site architecture and highlight the different structural solutions developed by nature for shells of marine organisms that serve the same purpose.

Pores and cracks act as stress concentrators during loading, making them a primary source of mechanical failure in brittle crystals. Remarkably, eggshells—composed of calcite, a material known for its brittleness—contain a high density of submicron closed voids, yet still act as an effective mechanical barrier that protects the embryo. By analyzing the void geometry and deformation behavior, we demonstrate that voids physically and geometrically serve as an essential structural motif with critical mechanical functions. The high density of intracrystalline voids, together with their disordered arrangement, inhibits lattice cleavage, hinders long‐range crack propagation, and promotes deformation twinning and quasi‐plasticity. The voids remain within a narrow subcritical size range, such that they predominantly do not act as crack initiators. Instead, through their size and spacing, they function as crack stoppers, thereby toughening the shell structure. Together, these voids play a crucial role in the mechanical integrity of ultrathin eggshells, which are primarily subjected to bending deformation. Our findings highlight the critical role of the size, density, and spatial distribution of isolated voids in thin‐walled avian eggshells and open new directions for designing flaw‐tolerant architectured single crystals and void‐mediated toughening strategies for thin‐film ceramics.