Conservation of Ornamental Stone by Myxococcus xanthus-Induced Carbonate Biomineralization

Increasing environmental pollution in urban areas has been endangering the survival of carbonate stones in monuments and statuary for many decades. Numerous conservation treatments have been applied for the protection and consolidation of these works of art. Most of them, however, either release dangerous gases during curing or show very little efficacy. Bacterially induced carbonate mineralization has been proposed as a novel and environmentally friendly strategy for the conservation of deteriorated ornamental stone. However, the method appeared to display insufficient consolidation and plugging of pores. Here we report that Myxococcus xanthus-induced calcium carbonate precipitation efficiently protects and consolidates porous ornamental limestone. The newly formed carbonate cements calcite grains by depositing on the walls of the pores without plugging them. Sonication tests demonstrate that these new carbonate crystals are strongly attached to the substratum, mostly due to epitaxial growth on preexisting calcite grains. The new crystals are more stress resistant than the calcite grains of the original stone because they are organic-inorganic composites. Variations in the phosphate concentrations of the culture medium lead to changes in local pH and bacterial productivity. These affect the structure of the new cement and the type of precipitated CaCO₃ polymorph (vaterite or calcite). The manipulation of culture medium composition creates new ways of controlling bacterial biomineralization that in the future could be applied to the conservation of ornamental stone.     

Influence of substrate mineralogy on bacterial mineralization of calcium carbonate: implications for stone conservation

The influence of mineral substrate composition and structure on bacterial calcium carbonate productivity and polymorph selection was studied. Bacterial calcium carbonate precipitation occurred on calcitic (Iceland spar single crystals, marble, and porous limestone) and silicate (glass coverslips, porous sintered glass, and quartz sandstone) substrates following culturing in liquid medium (M-3P) inoculated with different types of bacteria (Myxococcus xanthus, Brevundimonas diminuta, and a carbonatogenic bacterial community isolated from porous calcarenite stone in a historical building) and direct application of sterile M-3P medium to limestone and sandstone with their own bacterial communities. Field emission scanning electron microscopy (FESEM), atomic force microscopy (AFM), powder X-ray diffraction (XRD), and 2-dimensional XRD (2D-XRD) analyses revealed that abundant highly oriented calcite crystals formed homoepitaxially on the calcitic substrates, irrespective of the bacterial type. Conversely, scattered spheroidal vaterite entombing bacterial cells formed on the silicate substrates. These results show that carbonate phase selection is not strain specific and that under equal culture conditions, the substrate type is the overruling factor for calcium carbonate polymorph selection. Furthermore, carbonate productivity is strongly dependent on the mineralogy of the substrate. Calcitic substrates offer a higher affinity for bacterial attachment than silicate substrates, thereby fostering bacterial growth and metabolic activity, resulting in higher production of calcium carbonate cement. Bacterial calcite grows coherently over the calcitic substrate and is therefore more chemically and mechanically stable than metastable vaterite, which formed incoherently on the silicate substrates. The implications of these results for technological applications of bacterial carbonatogenesis, including building stone conservation, are discussed.     

Bioconservation of Deteriorated Monumental Calcarenite Stone and Identification of Bacteria with Carbonatogenic Activity

The deterioration of the stone built and sculptural heritage has prompted the search and development of novel consolidation/protection treatments that can overcome the limitations of traditional ones. Attention has been drawn to bioconservation, particularly bacterial carbonatogenesis (i.e. bacterially induced calcium carbonate precipitation), as a new environmentally friendly effective conservation strategy, especially suitable for carbonate stones. Here, we study the effects of an in situ bacterial bioconsolidation treatment applied on porous limestone (calcarenite) in the sixteenth century San Jeronimo Monastery in Granada, Spain. The treatment consisted in the application of a nutritional solution (with and without Myxococcus xanthus inoculation) on decayed calcarenite stone blocks. The treatment promoted the development of heterotrophic bacteria able to induce carbonatogenesis. Both the consolidation effect of the treatment and the response of the culturable bacterial community present in the decayed stone were evaluated. A significant surface strengthening (consolidation) of the stone, without altering its surface appearance or inducing any detrimental side effect, was achieved upon application of the nutritional solution. The treatment efficacy was independent of the presence of M. xanthus (which is known as an effective carbonatogenic bacterium). The genetic diversity of 116 bacterial strains isolated from the stone, of which 113 strains showed carbonatogenic activity, was analysed by repetitive extragenic palindromic–polymerase chain reaction (REP-PCR) and 16S rRNA gene sequencing. The strains were distributed into 31 groups on the basis of their REP-PCR patterns, and a representative strain of each group was subjected to 16S rRNA gene sequencing. Analysis of these sequences showed that isolates belong to a wide variety of phylogenetic groups being closely related to species of 15 genera within the Proteobacteria, Firmicutes and the Actinobacteria. This study shows that the abundant carbonatogenic bacteria present in the decayed stone are able to effectively consolidate the degraded stone by producing new calcite (and vaterite) cement if an adequate nutritional solution is used. The implications of these results for the conservation of cultural heritage are discussed.     

Bacterial Community Dynamics During the Application of a Myxococcus xanthus-Inoculated Culture Medium Used for Consolidation of Ornamental Limestone

In this study, we investigated under laboratory conditions the bacterial communities inhabiting quarry and decayed ornamental carbonate stones before and after the application of a Myxococcus xanthus-inoculated culture medium used for consolidation of the stones. The dynamics of the community structure and the prevalence of the inoculated bacterium, M. xanthus, were monitored during the time course of the consolidation treatment (30 days). For this purpose, we selected a molecular strategy combining fingerprinting by denaturing gradient gel electrophoresis (DGGE) with the screening of eubacterial 16S rDNA clone libraries by DGGE and sequencing. Quantification of the inoculated strain was performed by quantitative real-time PCR (qPCR) using M. xanthus-specific primers designed in this work. Results derived from DGGE and sequencing analysis showed that, irrespective of the origin of the stone, the same carbonatogenic microorganisms were activated by the application of a M. xanthus culture. Those microorganisms were Pseudomonas sp., Bacillus sp., and Brevibacillus sp. The monitoring of M. xanthus in the culture media of treated stones during the time course experiment showed disparate results depending on the applied technique. By culture-dependent methods, the detection of this bacterium was only possible in the first day of the treatment, showing the limitation of these conventional techniques. By PCR-DGGE analysis, M. xanthus was detected during the first 3–6 days of the experiment. At this time, the population of this bacterium in the culture media varied between 10⁸–10⁶ cells ml⁻¹, as showed by qPCR analyses. Thereafter, DGGE analyses showed to be not suitable for the detection of M. xanthus in a mixed culture. Nevertheless, qPCR analysis using specific primers for M. xanthus showed to be a more sensitive technique for the detection of this bacterium, revealing a population of 10⁴ cells ml⁻¹ in the culture media of both treated stones at the end of the consolidation treatment. The molecular strategy used in this study is proposed as an effective monitoring system to evaluate the impact of the application of a bacterially induced carbonate mineralization as restoration/conservation treatment for ornamental stones.     

不同培养条件对胶质芽孢杆菌诱导碳酸钙晶体形成的影响

【目的】研究不同培养条件对胶质芽孢杆菌(Bacillus mucilaginosus)菌体形态、数量和分泌的碳酸酐酶(CA酶)活性的影响,以及不同方式培养的菌体与碳酸钙晶体的生长及其形貌、数量之间的联系。【方法】分别采用无氮和有氮培养基培养胶质芽孢杆菌,进行菌体形态、数量及CA酶活性的比较,收集不同培养方式的菌体加入碳酸钙结晶体系中以研究细菌与碳酸钙晶体形成的联系。【结果】在无氮培养条件下,胶质芽孢杆菌数量少、荚膜肥厚,细菌培养液CA酶活力较低;有氮培养条件下,菌体数量多、荚膜单薄,细菌培养液CA酶活力较高。在碳酸钙结晶体系中加入无氮培养的菌体,生成的碳酸钙晶体表面光滑,体积较大但数量较小,加入有氮条件下培养的菌体形成的碳酸钙晶体表面粗糙,数量大但体积较小。【结论】不同培养条件能够引起胶质芽孢杆菌菌体数量、荚膜多糖及CA酶活的明显差异,进而对碳酸钙晶体的生成和形貌产生影响。     

Potential soil reinforcement by biological denitrification

Currently new ground reinforcement techniques are being developed based on microbially induced carbonate precipitation (MICP). Many studies on MICP use microbially catalyzed hydrolysis of urea to produce carbonate. In the presence of dissolved calcium this process leads to precipitation of calcium carbonate crystals, which form bridges between the sand grains and hence increase strength and stiffness. In addition to urea hydrolysis, there are many other microbial processes which can lead to the precipitation of calcium carbonate. In this study the theoretical feasibility of these alternative MICP processes for ground reinforcement is evaluated. Evaluation factors are substrate solubility, CaCO₃ yield, reaction rate and type and amount of side-product. The most suitable candidate as alternative MICP method for sand consolidation turned out to be microbial denitrification of calcium nitrate, using calcium salts of fatty acids as electron donor and carbon source. This process leads to calcium carbonate precipitation, bacterial growth and production of nitrogen gas and some excess carbon dioxide. The feasibility of MICP by denitrification is tested experimentally in liquid batch culture, on agar plate and in sand column experiments. Results of these experiments are presented and discussed.     

Nuclei plug septal pores in severed hyphae of Sordaria brevicollis

Colonies of Sordaria brevicollis cut with a razor blade were examined and compared to undamaged control colonies using light and transmission electron microscopy. Cut hyphae lost cytoplasm from severed compartments but retained cytoplasm in adjacent compartments due to the plugging of septal pores by nuclei. Hexagonal crystals were observed in hyphae but were neither positioned near to septal pores nor observed plugging them. Approximately 36% of setpal pores in undamaged hyphae were found to contain a nucleus, presumably migrating through them. It is suggested that nuclei plug septal pores in severed hyphae of S. brevicollis because they are more conveniently positioned to do so than the distant hexagonal crystals.     

Influence of Environmental Temperature on Biocalcification by Non-sporing Freshwater Bacteria

The process of biocalcification, recognised as precipitation of calcium carbonate, has been described as a widespread phenomenon associated with a wide range of different bacterial species. This biocalcifying activity, and factors that affect it, have been widely studied in moderately halophilic bacteria but there is a lack of information on factors that affect biocalcification by freshwater bacteria. In this paper, we study how temperature can affect biocalcification by freshwater bacteria that potentially could be used for the process of bioconsolidation during conservation. Ten isolates were characterised by standard biochemical and API 20NE tests. Their biocalcifying activity was studied at temperatures between 10 and 40°C in B4 liquid medium. Mineralogical and quantitative analyses of the crystals were carried out by XRD, and morphological studies by SEM. Biocalcification only occurred when bacteria were present and were able to grow. Carbonate precipitation by bacteria increased with time and temperature of incubation. Temperature affected not only the amount of precipitation but also crystal quality and morphology. As bioconsolidant agents, these organisms could be applied to stone when the temperature does not exceed 40°C depending on the type of isolate.     

The influence of culture conditions on growth and sheath development of calcifying cyanobacteria

Summary Calcifying cyanobacteria from the Everglades, Florida, USA, have been cultured in the laboratory. Nutrient concentration of the culture medium and illumination are of special importance for filaments physiologically and morphologically similar to the forms occurring in the natural habitat. High irradiance leads to a good development of an inner, pigmented, uncalcified sheath layer. When the cyano-bacteria grow in water supersaturated with respect to calcite, the outer sheath layers can be impregnated by carbonate crystals. The internal diameter of the resulting tube, however, depends on the-environmentally controlled-thickness of the uncalcified inner sheath.     

Ancient Fungal Life in North Pacific Eocene Oceanic Crust

We present evidence that eukaryotic life has existed in an extreme environment, inside the oceanic crust. Up to now only prokaryotes have been discovered within deep marine sediments and glass-rims of pillow basalts, no higher life forms are described as yet. This study demonstrates unique filamentous fossil structures observed within carbonate-filled vesicles of a massive lava flow unit from the upper oceanic crust in the North Pacific (ODP Site 1224). Based on morphological traits including branching, septa and central pores, the filaments are interpreted as fungi. The chemical composition of the fungal structures differs from the surrounding crystalline carbonate matrix in the deep basaltic rocks. Small open space between the fungi and the carbonate cement and undisturbed filamentous growth through different calcite crystals indicate endolithic fungal growth after the calcium carbonate filling. The presence of euhedral pyrite crystals within the carbonate cements points out anaerobic conditions in this habitat. Our results provide for the first time evidence for eukaryotic, fungal life in deep ocean basaltic rocks.     

Citrobacter sp. strain GW-M Mediates the Coexistence of Carbonate Minerals with Various Morphologies

To better understand the formation mechanism of carbonate minerals by microbes, culture experiments with a duration of 70 days were performed under the mediation of strain GW-M isolated from soil using modified Lagoa Vermelha (LV, a hypersaline coastal Lagoon, Rio de Janeiro, Brazil) medium with 6:1 Mg/Ca molar ratio. The results demonstrated that strain GW-M can mediate the formations of both high-Mg calcite and aragonite and that dumbbell-, cauliflower-, rhombohedra-shaped, and irregular minerals coexist in the modified LV medium. The amount of rhombohedra-shaped crystals increased significantly with culture time. A proposed mechanism for these formations is the following. Heterogeneous nucleation on the surface of the extracellular polymeric substances (EPS) always occurred, and carbonates with irregular shape existed in experimental products at any stages. The morphologies evolved from rod to dumbbell and finally to cauliflower. At the initial stage (till day 20), hydrogen ions and EPS secreted by the bacteria only influenced the microenvironment around the cells, and carbonates were precipitated on the surface of bacterial cells. At the middle and late stages (on days 45 and 70), microbes and their secretions influenced the whole medium. Under these conditions, rhombohedra-shaped crystals were formed when homogeneous nucleation occurred. In addition, the results of energy-dispersive spectrometry (EDS) showed that Mg contents in the synthesized carbonate minerals with rhombohedra-shaped were significantly lower than those of carbonates with other shapes, though relationship between morphology and species of mineral cannot be obtained by this phenomenon alone. These results shed further light on the mechanism of carbonate precipitation in the presence of microbes.     

Bacterial origin of the Red Pigmentation in the devonian Slivenec Limestone, Czech Republic

Summary The deep-red lenses of the Pragian Slivenec Limestone have been extensively quarried for ornamental purposes since the XII^th century. Petrographic microscope observations indicate that the hematite stainings of the limestone follow ten different patterns. They range from massive non-directional filling of cavities to mineralized films and microstromatolites. Numerous ironrich endolithes are observed. Some could be derived from bacterial or lichen perforations and some related to ferric bacteria. Infiltration along welded calcite crystals, regular mineralized films and microstromatolites suggest a ferric bacterial origin for the pigment. This is confirmed by scanning microscope examinations of polished sections, that show hematite concentrations along micrometric filamentous sheaths.     

Xanthan degragation by biofilm in porous media

Biological activity in oil reservoirs can cause significant problems such as souring and plugging. This study focuses on the problem of polymer degradation and permeability reduction due to biofilm formation during polymer injection for improved oil recovery. Polymers are included in injection fluids to increase their viscosity. Results relating biological processes and polymer degradation to fluid‐dynamic conditions in a laboratory model porous medium are presented.

A transparent flow cell with an etched two‐dimensional network of pores served as a model porous medium. A sterile xanthan polymer and natural sea water solution were continuously injected into the porous medium. A bacterial culture capable of xanthan degradation was introduced into the cell by a single injection. Some of the cells from this culture attached to the pore walls forming an immobile bacterial culture, termed biofilm. The development of this biofilm, its xanthan degradation and its effect on permeability were measured.

The effects of injection rate and rate transitions were analyzed. Injection fluid viscosity was reduced by 30% after 5 min flow through the porous medium at the maximum steady state degradation rate observed. Permeability was significantly reduced by the xanthan degrading biofilm, causing an increase in pressure drop through the porous medium of up to 80%. Polymer injection in oil reservoirs may, therefore, have negative effects on oil recovery, unless efficient biofouling control is applied. The methodology presented may serve as a tool in the development of biofouling control measures in porous media.     

Carbonate and Phosphate Precipitation by Chromohalobacter marismortui

The ability of Chromohalobacter marismortui to precipitate carbonate and phosphate minerals has been demonstrated for the first time. Mineral precipitation in both solid and liquid media at different salts concentrations and different magnesium/calcium ratios occurred whereas crystal formation was not observed in the control. The precipitated minerals were studied by X-ray diffraction, scanning electron microscopy and EDX, and were different in liquid and solid media. In liquid media aragonite, struvite, vaterite and monohydrocalcite were precipitated forming crystals and bioliths. Bioliths accreted preferentially close to organic pellicles, whereas struvite preferentially grows in microenvironments free of such pellicles. Magnesian calcite, calcian-magnesian kutnahorite, “proto-dolomite” and huntite were formed in solid media. The Mg content of the magnesian calcite and of Ca-Mg kutnahorite also varied depending on the salt concentration of the culture media. This is the first report on bacterial precipitation of Ca-Mg kutnahorite and huntite in laboratory cultures. The results of this research show the active role played by C. marismortui in mineral precipitation, and allow us to compare them with those obtained previously using other taxonomic groups of moderately halophilic bacteria.     

岩溶洞穴微生物沉积碳酸钙——以贵州石将军洞为例

为探讨洞穴微生物沉积碳酸钙作用对洞穴沉积物的影响,利用传统生物学方法,采集贵州中西部地区石将军洞洞穴沉积物表面的微生物样品,结合洞穴监测数据和理化背景资料,利用B-4培养基和B-4C培养基对洞穴细菌进行筛选和纯化,分离出能沉积碳酸钙的菌种,观察和了解洞穴细菌形成的CaCO3晶体,应用X射线衍射分析仪(XRD)分析细菌形成的CaCO3晶体成分,并利用扫描电子显微镜(SEM)观察晶体结构特征。结果表明:1)在B-4培养基下微生物产生的碳酸钙晶体主要为方解石、球霰石和方解石混合物、球霰石,这种变化与培养基pH值的增幅相关;同时,在添加Mg离子的B-4C培养基下形成的碳酸钙晶体主要为方解石,此外,研究中并未检测到文石晶体。2)通过SEM扫描,发现微生物作用形成的碳酸钙晶体存在不规则六方体、柱状体、四方体层状、半球状等,这些晶体形态在化学作用系统下少见,多见于微生物作用形成的方解石。此外,晶体中微生物作用痕迹明显,微生物作用贯穿于整个沉积过程。     

Surface analytical techniques applied to calcite: evidence of solid-state diffusion and implications for isotope methods

Surface analytical techniques have become common tools for physicists and materials scientists for making direct observations at the molecular scale but they can also provide Earth Scientists with new information about mineral/fluid interfaces. High resolution surface analysis compliments our traditional methods for analysing the bulk of solids and solutions which can give us, for example, a more complete picture of the geochemical processes that affect the mineral grains in sediments. Stable isotope methods have become widely used for dating and palaeoclimate studies but they rely on the assumption of a closed system. Artifacts resulting from carbonate mineral recrystallization can be avoided by careful sampling but ion by ion replacement can significantly alter the composition of a solid without leaving visible traces. Calcite has long been known to take up substituting ions into its atomic structure. Recent evidence gained from surface analysis shows Cd²⁺ and Zn²⁺, that had been adsorbed or precipitated on calcite surfaces, moves into calcite at the rate of nanometers per month, apparently by solid-state diffusion. No fractures that could serve as conduits from surface to bulk have ever been observed at the micrometer to nanometer scale on the single crystals used for these experiments. Mixing within the top few surface layers by reprecipitation during exposure to the humidity in air can account for some incorporation, but the evidence collected so far does not explain the exchange of position for ions beyond about ten calcite monolayers. With similar rates of movement, other trace components, such as K⁺, Na⁺, Cl^- and F^-, have been observed to move out of bulk calcite and to accumulate in discrete crystallites on surfaces exposed to air. Such mobility may be particular to these near-perfect, Iceland spar crystals and the ions investigated, but if O and C also move into and out of bulk calcite at similar rates, the integrity of isotope ratios from carbonate minerals, even from non-diagenetic environments, may be questionable.     

Soil Bioconsolidation Through Microbially Induced Calcite Precipitation by Lysinibacillus sphaericus WJ-8

Biomineralization is a process that leads to the formation of minerals via a biologically or biotechnologically mediated route. This process is a new and innovative research area in geotechnological engineering and structural engineering because it has wide-ranging implications for the strengthening of soil, sand, stone, and cementitious materials. In the present study, we demonstrated the ability of Lysinibacillus sphaericus WJ-8 to precipitate 15.3 mg/mL of calcite and to degrade 415 μmol/mL of urea over a 120-h period. The cell surface hydrophobicity and sand adhesion of spores were higher than those of vegetative cells (77.2% vs. 24.0% and 54.1% vs. 7.8%, respectively). In addition, the bioconsolidated soil block samples had significantly smaller pores than did the control soil block samples. Scanning electron microscopy and energy dispersive spectroscopy analysis revealed that calcite crystals were frequently formed in the bioconsolidated soil block samples, but did not occur in the control soil block samples. In addition, sharp peaks in the X-ray diffraction spectra indicated that calcite (CaCO₃) crystals constituted the predominant mineral in the bioconsolidated samples, whereas quartz (SiO₂) crystals constituted the predominant mineral in the control samples.     

Scanning electron microscopic and X-ray diffraction studies of otoconia in the lizard Podarcis s. sicula

Summary The otoliths of embryos and young animals of the lizard Podarcis s. sicula were studied by X-ray diffraction and scanning electron microscopy. Two types of crystal that give different X-ray diffraction patterns were found in the membranous labyrinth of Podarcis. The crystals consist of calcite or aragonite and are easily distinguished by scanning electron microscopy because of their different morphology. The two calcium carbonate crystal forms are not mixed at random but are present in the embryo from the very beginning in specific sites. The endolymphatic sac contains aragonite crystals while the saccule contains calcite crystals adjacent to the wall, in addition to a preponderance of aragonite crystals. The utricle and lagena contain only calcite crystals. The presence of two crystal forms of calcium carbonate in the membranous labyrinth are discussed in terms of differing genetic and functional significance.     

Reactive Transport Modeling of Induced Selective Plugging by Leuconostoc Mesenteroides in Carbonate Formations

A process-based mechanistic reactive transport model was developed to understand how in-situ coupled processes and operational factors affect selective plugging of reactive carbonate formations by the fermenting bacteria Leuconostoc mesenteroides that produces a plugging polymer dextran. The growth and transport of L. mesenteroides and the associated (bio) geochemical reactions were simulated explicitly with enzyme activity at the field scale over spatial extents of hundreds of meters. Simulations were performed to explore controls on selective bioplugging of high permeability zones in a representative carbonate reservoir, a process that can be used to improve oil sweep efficiency through lower permeability layers. Simulation results indicate that dextran production and the effectiveness of plugging can be largely affected by sucrose and bacteria injection rates. Selective plugging of high permeability zones can only be achieved when the injection rates are high compared to the rates of dextran production. Otherwise, plugging only occurs at the vicinity of injection wells. Due to the dependence of enzyme activity on pH and the reactive nature of carbonate formations, the chemistry of the injection and the formation water is also important. The injection of sucrose and L. mesenteroides at the optimum pH for dextran production (5.2) leads to the dissolution of calcite and an increase in pH levels. However, the resulting pH does not suppress plugging with dextran. Lactic acid and CO₂ formed during the growth of L. mesenteroides buffers the pH of water to levels between 5.2 and 7.0 for continued dextran production. At neutral and basic pH levels, induced precipitation of calcite does not significantly modify the permeability profile at carbonate concentrations typically found in oilfield formation waters. This is the first work that examines the controlling parameters that affect selective plugging of carbonate formations at the field scale within the context of enhanced oil recovery. The demonstrated approach can be used to identify optimal operational conditions for enhanced oil recovery and other applications where selective plugging can be beneficial.     

Influence of zinc on the calcium carbonate biomineralization of Halomonas halophila

Background

The salt tolerance of halophilic bacteria make them promising candidates for technical applications, like isolation of salt tolerant enzymes or remediation of contaminated saline soils and waters. Furthermore, some halophilic bacteria synthesize inorganic solids resulting in organic–inorganic hybrids. This process is known as biomineralization, which is induced and/or controlled by the organism. The adaption of the soft and eco-friendly reaction conditions of this formation process to technical syntheses of inorganic nano materials is desirable. In addition, environmental contaminations can be entrapped in biomineralization products which facilitate the subsequent removal from waste waters. The moderately halophilic bacteria Halomonas halophila mineralize calcium carbonate in the calcite polymorph. The biomineralization process was investigated in the presence of zinc ions as a toxic model contaminant. In particular, the time course of the mineralization process and the influence of zinc on the mineralized inorganic materials have been focused in this study.

Results

H. halophila can adapt to zinc contaminated medium, maintaining the ability for biomineralization of calcium carbonate. Adapted cultures show only a low influence of zinc on the growth rate. In the time course of cultivation, zinc ions accumulated on the bacterial surface while the medium depleted in the zinc contamination. Intracellular zinc concentrations were below the detection limit, suggesting that zinc was mainly bound extracellular. Zinc ions influence the biomineralization process. In the presence of zinc, the polymorphs monohydrocalcite and vaterite were mineralized, instead of calcite which is synthesized in zinc-free medium.

Conclusions

We have demonstrated that the bacterial mineralization process can be influenced by zinc ions resulting in the modification of the synthesized calcium carbonate polymorph. In addition, the shape of the mineralized inorganic material is chancing through the presence of zinc ions. Furthermore, the moderately halophilic bacterium H. halophila can be applied for the decontamination of zinc from aqueous solutions.