Biofilm Medium Chemistry and Calcium Oxalate Morphogenesis.

The present study is focused on the effect of biofilm medium chemistry on oxalate crystallization and contributes to the study of the patterns of microbial biomineralization and the development of nature-like technologies, using the metabolism of microscopic fungi. Calcium oxalates (weddellite and whewellite in different ratios) were synthesized by chemical precipitation in a weakly acidic environment (pH = 4-6), as is typical for the stationary phase of micromycetes growth, with a ratio of Ca2+/C2O42- = 4.0-5.5, at room temperature. Additives, which are common for biofilms on the surface of stone in an urban environment (citric, malic, succinic and fumaric acids; and K+, Mg2+, Fe3+, Sr2+, SO42+, PO43+ and CO32+ ions), were added to the solutions. The resulting precipitates were studied via X-ray powder diffraction (XRPD), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDXS). It was revealed that organic acids, excreted by micromicetes, and some environmental ions, as well as their combinations, significantly affect the weddellite/whewellite ratio and the morphology of their phases (including the appearance of tetragonal prism faces of weddellite). The strongest unique effect leading to intensive crystallization of weddellite was only caused by the presence of citric acid additive in the medium. Minor changes in the composition of the additive components can lead to significant changes in the weddellite/whewellite ratio. The effect of the combination of additives on this ratio does not obey the law of additivity. The content of weddellite in the systems containing a representative set of both organic acids and environmental ions is ~20 wt%, which is in good agreement with natural systems.

Characterization of Calcium Oxalate Hydrates and the Transformation Process.

Calcium oxalate can be found in humans as kidney stones and in cultural heritage as films in two crystallographic species, dihydrate (COD/weddellite) and/or monohydrate (COM/whewellite). Due to its instability, COD is transformed into COM. Studying this crystalline conversion provides information about the origin of the monohydrated species, which will help in the assessment of prevention measurements to avoid their formation. In the present study, the synthesis of calcium oxalate hydrate microcrystals has been carefully performed to avoid mixture of phases in the final products; the long and short range order structure of both species have been studied by X-ray diffraction (XRD) and X-ray absorption spectroscopy (XAS), respectively. This structural information was considered in the density functional theory (DFT) computational study performed to assign the characteristic vibrational IR and Raman frequencies found. This detailed characterization allows an unambiguous assignment of the vibrations, thus providing the appropriate parameters required to monitor and characterize the transformation process.

Scanning Electron Microscopy Coupled with Energy Dispersive Spectrometric Analysis Reveals for the First Time Weddellite and Sylvite Crystals on the Surface of Involucral Bracts and Petals of two Xeranthemum L. (Compositae) Species.

In this work, weddellite and sylvite crystals are identified for the first time on the involucral bracts and petals of Xeranthemum annuum and Xeranthemum cylindraceum using scanning electron microscopy coupled with energy dispersive spectrometric (SEM-EDS) analysis. Well-developed crystals of weddellite (CaC2O4·2H2O) occur in the form of a tetragonal bipyramid (hhl), rarely in combination of a bipyramid and tetragonal prism (h00). Indumentum of involucral bracts of X. cylindraceum consists of nonglandular and glandular trichomes. Sylvite (KCl) crystals are observed only on the petal surface of X. cylindraceum. The crystals of sylvite occur in the form of perfect cubes (hexahedrons), but some crystals are deformed, i.e., partially elongated. Taxonomic significance of investigated microcharacters as well as the use of SEM-EDS analysis in taxonomic studies of plants are discussed.

Abiotic Formation of Calcium Oxalate under UV Irradiation and Implications for Biomarker Detection on Mars.

A major objective in the exploration of Mars is to test the hypothesis that the planet has ever hosted life. Biogenic compounds, especially biominerals, are believed to serve as biomarkers in Raman-assisted remote sensing missions. However, the prerequisite for the development of these minerals as biomarkers is the uniqueness of their biogenesis. Herein, tetragonal bipyramidal weddellite, a type of calcium oxalate, is successfully achieved by UV-photolyzing pyruvic acid (PA). The as-prepared products are identified and characterized by micro-Raman spectroscopy and field emission scanning electron microscopy. Persistent mineralization of weddellite is observed with altering key experimental parameters, including pH, Ca2+ and PA concentrations. In particular, the initial concentration of PA can significantly influence the morphology of weddellite crystal. Oxalate acid is commonly of biological origin; thus calcium oxalate is considered to be a biomarker. However, our results reveal that calcium oxalate can be harvested by a UV photolysis pathway. Moreover, prebiotic sources of organics (e.g., PA, glycine, alanine, and aspartic acid) have been proven to be available through abiotic pathways. Therefore, our results may provide a new abiotic pathway of calcium oxalate formation. Considering that calcium oxalate minerals have been taken as biosignatures for the origin and early evolution of life on Earth and astrobiological investigations, its formation and accumulation by the photolysis of abiological organic compounds should be taken into account.

Variability and composition of calcium oxalate crystals in embryos-seedlings-adult plants of the globose cacti Mammillaria uncinata.

The morphology and elemental composition of crystals in embryos, seedlings and adult plants of the globose cacti Mammillaria uncinata were studied. Samples of mature fruits and adult plants were collected. To obtain embryos and seedlings, seeds from mature fruits were germinated under laboratory conditions. Both embryos and seedlings as well as stem and root of the adult plants were processed by conventional microtechniques and tissue macerations to isolate the individual crystals. The crystal morphology was evaluated by light and scanning electron microscopy while its chemical composition was analyzed by energy dispersive spectroscopy, IR and Raman spectroscopy. The results demonstrated the occurrence of calcium oxalate crystals in the three stages of plant growth. Solitary and small crystals are present in embryos and seedlings in contrast crystals are aggregates or conglomerates in the adult plants. The compositional analysis showed that seedling crystals contain carbon (50.37%), oxygen (45.29%) and calcium (3.36%) while in adult plants the percentage has changed to carbon (12.54%), oxygen (53.06%) and calcium (34.38%). In the IR spectrum, the vibration bands around 1321 and 1621 cm-1 are attributed to the calcium oxalate in the dihydrate state (weddellite), the Raman peak at 1475 cm-1 shows also that crystals correspond to the pure state of calcium oxalate dihydrate state. The crystal size was also different for seedlings and adult plants, mean values varied from 12.11 to 13.38 µm for width and length, respectively in seedlings and from 65.10 to 73.90 µm, in adult plants. It is concluded that the elemental composition, size and morphology of crystals in M. uncinata depend on the growth stage, as it happens in other plant species.

CO2 capture and sequestration in stable Ca-oxalate, via Ca-ascorbate promoted green reaction.

The increase in the amount of carbon dioxide (CO2) emissions related to many anthropic activities is a persistent and growing problem. During the last years, many solutions have been set out, none of them being the ultimate one. Investigators agree on the need of a synergic approach to the problem, in terms of many complementary methods of sequestration that, combined with the reduction of production, will be able to decrease the concentration of the CO2 in the atmosphere. In this work, we explore the use of a green reaction to trap the CO2 into a stable crystalline phase (weddellite) resorting to a multidisciplinary approach. CO2 is reduced and precipitated as calcium oxalate through vitamin C as a sacrificial reductant. Calcium oxalate crystals obtained show a startling good quality that increases their already great stability over a wide chemical and physical conditions' range.

Biogenesis of secondary mycogenic minerals related to wall paintings deterioration process.

Present study addresses potential of fungal strains, isolated from deteriorated mural paintings and surrounding air environment of the Church of the Holy Ascension in Veliki Krcimir (Serbia), to precipitate mycogenic minerals, when cultivated on agarized B4 medium. Utilizing culture-based isolation methods, 38 filamentous fungi were obtained in total, 23 from mural paintings and 15 from air, respectively, mainly ascomycetes, while Bjerkandera adusta and Thanatephorus cucumeris were only basidiomycetes. A total of 31 of 38 fungal isolates, more than 80%, were able to form minerals of different morphologies and variable size, determined via SEM-EDS and XRPD, to be either calcite or calcite and weddellite association. Among screened fungi, all Penicillium, Chaetomium and Cladosporium isolates, as well as most of the Aspergillus isolates (8/11) precipitated minerals, whereas cultures of Bionectria, Bjerkandera, and Seimatosporium isolates lacked any observable crystal forms. With the exception of two Alternaria alternata strains, no apparent disparity in potential to precipitate minerals in general, or form particular crystal phase was documented among the air and mural paintings isolates. Possible mechanisms of fungal mineralization of calcite and weddellite are further proposed. In addition to providing experimental evidence for fungal induced precipitation of oxalate and carbonate minerals, presented data suggest that fungal activity could be an important factor in a weathering process affecting cultural heritage exhibited and stored in inadequate conditions. Implementation of B4 plate assay for screening of mineralization potential of the isolated fungi could be used to assess biodegradative risk mycobiota pose to the mural paintings, so appropriate conservation measures may be utilized.

Phase transformation of calcium oxalate dihydrate-monohydrate: effects of relative humidity and new spectroscopic data.

New data on vibrational properties of calcium oxalates and their controversial transformation mechanism are presented. We have focused on whewellite (CaC2O4·H2O) and weddellite [CaC2O4·(2+x) H2O], the most common phases of calcium oxalate; these compounds occur in many organisms, in kidney stones and in particular kinds of films found on the surface of many works of art. Low temperature experiments carried out by Fourier transform infrared spectroscopy have highlighted both the high structural order in the crystalline state of whewellite and the disordered distribution of the zeolitic water molecules in weddellite. The synthesised nanocrystals of weddellite have been kept under different hygrometric conditions in order to study, by X-ray powder diffraction, the role of "external" water molecules on their stability. Moreover, in order to identify the different kinds of water molecules, a re-investigation, supported by quantum chemical calculations, of the observed vibrational spectra (IR and Raman) of whewellite has been conducted.