Maggie Cusack

Structure, composition and mechanical relations to function in sea urchin spine

Sea urchins have characteristic spines that fulfil critical functions. Several studies revealed marked spine internal heterogeneities at different structural levels despite the single-crystal character of the spines. Most of these studies did not speculate about the functional meaning of these heterogeneities. Spine heterogeneities were investigated in the sea urchin Paracentrotuslividus and their possible functional implications discussed. Spines mainly show two morphological parts: the base, made of a meshwork stereom, and the shaft, with longitudinal plain septa and a central core of meshwork stereom. Electron Backscatter Diffraction showed no difference in crystallite orientation between the two structures. Atomic Absorption Spectrometry and Energy dispersive X-ray analysis revealed that Mg was not uniformly distributed in the spine. Mg concentration is higher in the inner part of the septa than in the septum outer part. Furthermore, a cyclic pattern of Mg concentration in septa was observed. This is suggested to be linked to the spine ontogeny. Nano- and microindentation analyses revealed that the septa have higher stiffness and hardness than the meshwork stereom and that septum stiffness and hardness present different trends in longitudinal and transverse section. These mechanical heterogeneities may have an adaptive functional value.

Crystallography and chemistry of the calcium carbonate polymorph switch in M. edulis shells

The control exerted by some invertebrates on the calcium carbonate polymorph produced is intriguing but not understood. Mytilus edulis shells, with the abrupt polymorph switch within their valves from an outer calcite to inner aragonite layer, are excellent examples of this phenomenon. Detailed crystallography of intact valves using Electron Backscatter Diffraction (EBSD) is considered in the context of quantitative chemical analyses by electron microprobe. Apart from the outer 40 μm, individual crystals that comprise the calcite layer of M. edulis differ from each other in terms of misorientation by less than 10°. Similar uniformity occurs in the inner aragonite layer with notable ‘mineral bridging’ between tablets of aragonite nacre. The first-formed aragonite laminae are sub-micron thickness and the subsequent laminae of uniform 1 μm thickness. Variations in chemical composition through the two valves correspond in part with the distribution of the two polymorphs. Magnesium is present in notably higher concentrations within calcite than aragonite. However, the Mg2+ concentration in calcite is not uniform and increases with growth before decreasing at the polymorph switch. Sodium concentrations decrease steadily through the calcite layer. The aragonite layer is compositionally more uniform. Sulphur is not a good proxy for organic content in this system because it does not reflect the higher organic content of the aragonite. Sector zoning is not responsible for the element distribution seen here while differences in crystal size and association with organic components remain as possible explanations.

Chemico‐structural evolution of linguloid brachiopod shells

Chemico-structures of shells representing all families presently assigned to the Linguloidea have undergone significant transformations since the Early Cambrian. Superficial hemispherical to hemi-ellipsoidal pits on the larval and/or mature shells are interpreted as casts of deformable, membrane-bound vesicles of mucus or rigid vesicles of glycoproteins or GAGs with thickened coats. Flat-bottomed, sub-circular imprints characterize acrotheloids and many acrotretides, and could be impressions of biconvex tablets of apatite like those exocytosed within the primary layer of the obolid ‘Lingulella’? antiquissima, whilst the rhomboidal imprints of the Paterula shell could have held tablets of proteinaceous silica like those of living discinid larvae. The ancestral fabric of the linguloid secondary layer was probably composed of rubbly and virgose sets, but trellised rods of apatite (baculation) are characteristic of most linguloids and also acrotheloids. This condition was suppressed in shells identified as ‘Lingula’ from at least the Early Carboniferous to the present day. In early Palaeozoic acrotretides and lingulellotretids, columnar and camerate fabrics evolved in place of baculation. Baculation in Discinisca tenuis and Glottidia pyramidata is associated with the amino acids glutamic acid, glycine, alanine, arginine and proline which may be components of an organic polymer axial to baculate accretion.

Aragonite-calcite seas—Quantifying the gray area

Oscillations between the dominance of aragonite and calcite in abiotic marine CaCO3 precipitates throughout Earth history are closely coupled with the evolution of Earth’s seawater composition and represent the environmental context in which organisms evolved their ability to biomineralize. The most important factor controlling these Phanerozoic oscillations in CaCO3 polymorph composition is the ratio of Mg:Ca in seawater, which is thought to separate aragonite and calcite precipitation along a distinct temperature-controlled threshold. A sharp threshold, however, is contradicted by overlapping aragonite and calcite precipitation fields at a range of experimental conditions. Here we present experimental data that enable us to quantify the proportions of CaCO3 polymorphs as a function of Mg:Ca ratio and temperature. This allows us to convert published Mg:Ca ratio proxy data and models of the Phanerozoic Mg:Ca ratio into proportions of abiotic CaCO3 polymorphs at a given temperature, and thus provides a temperature-corrected view of aragonite-calcite sea conditions. In this revised view, abiotic calcite precipitation was inhibited during aragonite sea intervals at temperatures above 20 °C, whereas calcite sea intervals were characterized by the co-precipitation of aragonite and calcite in environments above 20 °C. This continuous prominence of aragonite precipitation in Phanerozoic warm-water environments explains the Phanerozoic increase of aragonite over calcite skeletal composition in calcifying organisms.

Ocean acidification impacts mussel control on biomineralisation

Ocean acidification is altering the oceanic carbonate saturation state and threatening the survival of marine calcifying organisms. Production of their calcium carbonate exoskeletons is dependent not only on the environmental seawater carbonate chemistry but also the ability to produce biominerals through proteins. We present shell growth and structural responses by the economically important marine calcifier Mytilus edulis to ocean acidification scenarios (380, 550, 750, 1000 µatm pCO2). After six months of incubation at 750 µatm pCO2, reduced carbonic anhydrase protein activity and shell growth occurs in M. edulis. Beyond that, at 1000 µatm pCO2, biomineralisation continued but with compensated metabolism of proteins and increased calcite growth. Mussel growth occurs at a cost to the structural integrity of the shell due to structural disorientation of calcite crystals. This loss of structural integrity could impact mussel shell strength and reduce protection from predators and changing environments.

Ocean acidification reduces the crystallographic control in juvenile mussel shells.

Global climate change threatens the oceans as anthropogenic carbon dioxide causes ocean acidification and reduced carbonate saturation. Future projections indicate under saturation of aragonite, and potentially calcite, in the oceans by 2100. Calcifying organisms are those most at risk from such ocean acidification, as carbonate is vital in the biomineralisation of their calcium carbonate protective shells. This study highlights the importance of multi-generational studies to investigate how marine organisms can potentially adapt to future projected global climate change. Mytilus edulis is an economically important marine calcifier vulnerable to decreasing carbonate saturation as their shells comprise two calcium carbonate polymorphs: aragonite and calcite. M. edulis specimens were cultured under current and projected pCO2 (380, 550, 750 and 1000μatm), following 6months of experimental culture, adults produced second generation juvenile mussels. Juvenile mussel shells were examined for structural and crystallographic orientation of aragonite and calcite. At 1000μatm pCO2, juvenile mussels spawned and grown under this high pCO2 do not produce aragonite which is more vulnerable to carbonate under-saturation than calcite. Calcite and aragonite were produced at 380, 550 and 750μatm pCO2. Electron back scatter diffraction analyses reveal less constraint in crystallographic orientation with increased pCO2. Shell formation is maintained, although the nacre crystals appear corroded and crystals are not so closely layered together. The differences in ultrastructure and crystallography in shells formed by juveniles spawned from adults in high pCO2 conditions may prove instrumental in their ability to survive ocean acidification.

Material properties of brachiopod shell ultrastructure by nanoindentation

Mineral-producing organisms exert exquisite control on all aspects of biomineral production. Among shell-bearing organisms, a wide range of mineral fabrics are developed reflecting diverse modes of life that require different material properties. Our knowledge of how biomineral structures relate to material properties is still limited because it requires the determination of these properties on a detailed scale. Nanoindentation, mostly applied in engineering and materials science, is used here to assess, at the microstructural level, material properties of two calcite brachiopods living in the same environment but with different modes of life and shell ultrastructure. Values of hardness (H) and the Young modulus of elasticity (E) are determined by nanoindentation. In brachiopod shells, calcite semi-nacre provides a harder and stiffer structure (H∼3–6 GPa; E=60–110/120 GPa) than calcite fibres (H=0–3 GPa; E=20–60/80 GPa). Thus, brachiopods with calcite semi-nacre can cement to a substrate and remain immobile during their adult life cycle. This correlation between mode of life and material properties, as a consequence of ultrastructure, begins to explain why organisms produce a wide range of structures using the same chemical components, such as calcium carbonate.

High resolution electron backscatter diffraction (EBSD) data from calcite biominerals in recent gastropod shells

Electron backscatter diffraction (EBSD) is a microscopy technique that reveals in situ crystallographic information. Currently, it is widely used for the characterization of geological materials and in studies of biomineralization. Here, we analyze high resolution EBSD data from biogenic calcite in two mollusk taxa, Concholepas and Haliotis, previously used in the understanding of complex biomineralization and paleoenvironmental studies. Results indicate that Concholepas has less ordered prisms than in Haliotis, and that in Concholepas the level of order is not homogenous in different areas of the shell. Overall, the usefulness of data integration obtained from diffraction intensity and crystallographic orientation maps, and corresponding pole figures, is discussed as well as its application to similar studies.

Relic aragonite from Ordovician-Silurian brachiopods: Implications for the evolution of calcification

Understanding the influence of aragonite/calcite sea conditions on the evolution of biocalcification relies strongly on the correct interpretation of the original composition of calcareous taxa. Aragonite dissolves or inverts into calcite over geological time, and its preservation is currently unknown to predate the Pennsylvanian. Here we present direct evidence for the common occurrence of relic aragonite in Ordovician and Silurian trimerellid brachiopods, thereby extending the known range of aragonite preservation by more than 130 million years. Together with associated hypercalcifying taxa of putatively original aragonite or high-magnesium calcite composition and considerations of the temperature dependence of aragonite and calcite precipitation, our results suggest that the evolution of aragonite biomineralization might have presented an adaptive advantage in shallow marine tropical waters of calcite seas. A targeted search for Paleozoic aragonite should both resolve the original composition of consistently recrystallized taxa and enable the reassessment of the aragonite/calcite sea paradigm in a paleoenvironmental context.

Magnesium and phosphorus distribution in the avian eggshell.

Magnesium and phosphorus are major inorganic constituents of the avian eggshell. The Mg/Ca ratio has been used as a palaeothermometer in a range of calcite biominerals. Eggshells provide the opportunity to examine the Mg/Ca ratio of a calcite biomineral produced in a constant temperature environment. Mg distribution is not constant throughout the shell, decreasing from nucleation until after fusion of the mammillary caps and then increasing to termination. This indicates that temperature of deposition is not the only factor controlling the Mg content of this biomineral system. There is a greater increase in magnesium concentration in the outer region of eggshells from older birds. The variation in magnesium concentration does not appear to correlate with organic content. Phosphorus occurs in the outer quarter of the eggshell and rises to termination and is therefore not confined to cuticular vesicles.