No abstract

This work presents a detailed structural and morphological analysis of different dinosaur eggshells such as Spheroolithus (sample 1, 2), lambeosaurinae, Prismatoolithus , and one unidentified ootaxon performed by high-resolution scanning electron microscopy (HRSEM). These ancient eggshells of Late Cretaceous dinosaurs were collected in the coastal area of El Rosario, Baja California in Mexico. Additionally, a thorough study was performed on the elements present in the samples by different techniques such as energy-dispersive spectroscopy (EDS), X-ray fluorescence (XRF), and X-ray photoelectron spectroscopy (XPS). The XPS technique was performed to make an accurate identification of the compounds of two different types of eggshells ( Spheroolithus sample 1 and Prismatoolithus ). This contribution compares the surface of five different dinosaur eggshells of 74 Ma and their inner section to determine the morphology, distribution of the chemical elements present, as well as their relationship. The observed morphology of the ornithopod eggshells of the herbivorous species shows that the mammillary cones are in the form of columns with microaggregates and irregular pores. In contrast, in the theropod eggshells, the mammillary cones are observed in different forms with wider pores. Finally, the chemical components present in the structures of each of the samples were estimated using the information obtained from SEM-EDS, evidencing the presence of calcite, quartz, and albite in each of the samples. The composition reveals that eggshells contain Si, P, S, K, Ca, Mn, Fe, and Sr and trace elements such as Cr, Cu, and Zn. The presence of heavy metals may be an indication that the eggshells presented diagenetic alterations.

We performed a molecular analysis of dinosaur's eggshells collected in El Rosario, Baja California, using techniques such as Raman spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRPD), and photoluminescence spectroscopy (PS). These techniques allowed us to identify calcite as the main mineral. The crystalline planes corresponding to this mineral were obtained with a hexagonal unit cell by Rietveld refinement. In addition, six crystallographic planes corresponding to quartz were identified by XRPD, with a rather large crystallite size compared to those obtained from calcite and albite. For this latter mineral, found in a low content, we were able to identify only three crystalline planes and three wavelengths of photoluminescence and micro-Raman signals at a very low intensity. In addition, the analysis by FT-IR allowed us to identify bands corresponding to nine amino acids and the components of the secondary structure that might very well be some ancestral proteins that had been conserved for so long, thanks to the mineralized structure. This type of structural characterization together with the optical one is a very relevant contribution to the field of paleontological research, mainly because these types of samples are unique in their type due to the biological relevance in Mexico that will allow us to understand the species that became extinct millions of years ago.

In this work, we present an investigation of the surface area and roughness of different dinosaur eggshells of 70 million years old using fractal dimension analysis obtained from atomic force microscopy (AFM) and scanning electron microscopy (SEM) information. We also conduct qualitative analyses on the external and inner surfaces of eggshells, which are mainly composed of calcium carbonate. The morphological characteristics of both surfaces can be revealed by both SEM and AFM techniques. It is observed that the inner surface of the eggshell has greater roughness that increases the surface area due to the vaster number of pores compared to the external face, making, therefore, the fractal dimension also greater. The aim of this contribution is to identify the morphology of the pores, as well as the external and inner surfaces of the eggshells, since the morphology is very similar on both surfaces and will otherwise be difficult to determine with the naked eye by SEM and AFM. In addition, the sole AFM analysis is very complicated for these types of samples due to the intrinsic roughness. However, it needs additional methods or strategies to complete this purpose. This contribution used the fractal dimension to show the same behavior obtained in both SEM and AFM techniques, indicating the fractal nature of the structures.

In this talk, the role that intramineral proteins have played on the shape control as well as in the biomineralization of calcium carbonate in the eggshell´s formation of different avian, crocodiles and dinosaurs will be reviewed. Particularly, the collected eggshells samples of five fossilized eggshells from dinosaurs that roamed the Earth more than 65 million years ago. We characterized the eggshells of the Theropod (bipedal carnivores) and Hadrosauridae (duck-billed dinosaurs) families and an unidentified ootaxon. We have found the existence of some proteins by using micro X-ray absorption and micro-fluorescence techniques at the synchrotron facilities. From these analyses on the dinosaur eggshells, X-ray absorption methods showed a very characteristic organic sulfur bonding similar to that semi-essential proteogenic amino acid L-cysteine, which implies that there is a possibility of having a very old intramineral protein similar to those found in emu and crocodiles. On the other hand, the spectroscopical characterization on these samples showed that calcium carbonate was the primary mineral, with smaller amounts of albite and quartz crystals. Anhydrite, hydroxyapatite, and iron oxide impurities were also present in the shells, which suggests replacement of some of the original minerals during fossilization. Then, with Fourier transform infrared spectroscopy (FT-IR), we found nine amino acids among the five samples, being lysine the only amino acid present in all of them. In addition, we have found evidence of secondary protein structures, including turns, α-helices, β-sheets and disordered structures, which have been preserved for millions of years by being engrained in the minerals. The FT-IR bands corresponding to amino acids and secondary structures could be indicative of ancestral proteins that have not been characterized before. This type of chemical, spectroscopical and structural characterization together with the optical one is a relevant contribution to the field of biomineralization of calcium carbonate research, mainly because these types of samples are unique in their type due to the biological relevance in Mexico and will, therefore, allow us to understand the species that became extinct millions of years ago as well as the importance of calcium carbonate associated to ancient proteins throughout the biomineralization processes on Earth.