Spring-associated limestones of the Eastern Alps: overview of facies,deposystems, minerals,and biota

In the Eastern Alps, both fossil spring limestones and actively limestone-depositing springs are common. The geological context and a few radiometric age data of fossil spring-associated limestones (SAL) mentioned herein indicate that they accumulated subsequent to the Last Glacial Maximum in the Eastern Alps (24–21 ka BP). Prevalent facies of the SAL deposits, active and fossil, including phytoclastic tufa, microbialites s.l., springstone, and moss tufa form, or formed, from (a) waterfall/creek systems, (b) hillslope-paludal systems, (c) moss-tufa systems, and from (c) foreland-type systems. Precipitated minerals include calcite and, at springs of elevated Mg/Ca ratio, magnesian calcite and aragonite. In a few limestone-depositing, oxygen-deficient springs with dissolved Fe²⁺, downstream, iron oxide precipitates ahead of CaCO₃ (mineralogical zonation). Biota associated with calcium-carbonate deposition include cyanobacteria, green micro-algae, macro-algae, and mosses. Calcium-carbonate precipitation may be speeded by biological mediation, but mineralogy and polymorphy of precipitated CaCO₃ are not biotically controlled. In the Eastern Alps, SAL deposits in total range from 190 to 2,520 m a.s.l., corresponding to mean annual temperatures of 10°C to less than 0°C. In altitudes below the continuous permafrost line (about 2,600–3,000 m a.s.l., depending on location), SAL deposition is chiefly controlled by proper balance between water supply and sufficient supersaturation for CaCO₃, rather than by mean annual temperature.     

Flow Velocity Effect on Leaf Litter Breakdown in Tufa Depositing System (Plitvice Lakes,Croatia)

Considerable amount of riparian leaf litter is annually supplied to the cascade Plitvice Lakes and trapped on tufa barriers where it decays together with aquatic macrophytes. These barriers are the sites of heavy calcite precipitation that can widely differ in terms of current velocity. We conducted a leafbag experiment at sites differing in flow velocity and tufa deposition rate. Decomposition of Petasites spp. and Fagus sylvatica was higher under high current (0.80 m/s) and high tufa deposition areas than in low current (< 0.20 m/s) and low tufa deposition areas (k = 0.085 vs. 0.021 for Petasites spp. and 0.009 vs. 0.002 for Fagus sylvatica). We concluded that although tufa deposition could interfere with decomposition by obstructing physical abrasion and also restricting microbial conditioning, thin calcite crusts developed on the surface of the leaves made them more fragile and thus accelerated their decomposition. High current velocity probably magnified this effect by supporting higher tufa deposition and coarser type of tufa fabrics. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Potential role for microbial ureolysis in the rapid formation of carbonate tufa mounds

Modern carbonate tufa towers in the alkaline (~pH 9.5) Big Soda Lake (BSL), Nevada, exhibit rapid precipitation rates (exceeding 3 cm/year) and host diverse microbial communities. Geochemical indicators reveal that carbonate precipitation is, in part, promoted by the mixing of calcium-rich groundwater and carbonate-rich lake water, such that a microbial role for carbonate precipitation is unknown. Here, we characterize the BSL microbial communities and evaluate their potential effects on carbonate precipitation that may influence fast carbonate precipitation rates of the active tufa mounds of BSL. Small subunit rRNA gene surveys indicate a diverse microbial community living endolithically, in interior voids, and on tufa surfaces. Metagenomic DNA sequencing shows that genes associated with metabolisms that are capable of increasing carbonate saturation (e.g., photosynthesis, ureolysis, and bicarbonate transport) are abundant. Enzyme activity assays revealed that urease and carbonic anhydrase, two microbial enzymes that promote carbonate precipitation, are active in situ in BSL tufa biofilms, and urease also increased calcium carbonate precipitation rates in laboratory incubation analyses. We propose that, although BSL tufas form partially as a result of water mixing, tufa-inhabiting microbiota promote rapid carbonate authigenesis via ureolysis, and potentially via bicarbonate dehydration and CO₂ outgassing by carbonic anhydrase. Microbially induced calcium carbonate precipitation in BSL tufas may generate signatures preserved in the carbonate microfabric, such as stromatolitic layers, which could serve as models for developing potential biosignatures on Earth and elsewhere.     

Tufa Deposition in Karst Streams Can Enhance the Food Supply of the Grazing Caddisfly Melampophylax mucoreus (Limnephilidae)

We studied the effect of carbonate depositions covering stone surfaces on the growth of larvae and the biomass of subsequent adults of the grazing limnephilid caddisfly Melampophylax mucoreus (Hagen , 1861) in a laboratory rearing experiment. M. mucoreus is mainly distributed in karst streams characterized by calcium carbonate precipitations (tufa). We reared larvae of M. mucoreus on stones covered by calcareous tufa crusts as well as on stones from which these crusts were experimentally removed to assess the influence on larval growth and the subsequent adult biomass. The rough surface of the covered stones provided a higher complexity of micro‐habitats and supported algal growth compared to the smooth surface of stones without crusts. Larvae of M. mucoreus profited from the enhanced algal biofilm growth on the calcium carbonate precipitation indicated by faster larval growth and higher subsequent adult biomass. Biomass increase of larvae reared on stones covered by tufa crusts exhibited a faster biomass development (0.09 ± 0.015 mg/d) compared to the larvae reared on stones without crusts (0.06 ± 0.002 mg/d). Adult males (5.13 ± 0.25 mg) and females (7.64 ± 0.63 mg) were significantly heavier in the treatment with stones covered by tufa than their conspecifics from the treatment with uncovered stones (males: 4.26 ± 0.25 mg, p = 0.047; females: 4.96 ± 0.47 mg, p = 0.001). Additionally, males from the treatment with crust covered stones emerged significantly earlier (p = 0.003) than the males from the other treatment, whereas no significant difference was found for females. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Modern freshwater microbial carbonates: thePhormidium stromatolites (tufa-travertine) of southeastern Burgundy (Paris Basin,France)

Summary The relationships of microstructure and vegetal remains, obtained by decalcification, were studied in Modern tufa from Burgundy, in order to try to link a given species with a particular crystal habitus. The edifices have various shapes (coatings on floors; encrusted pebbles, shells, vegetal shoots, mosses; oncolites; hydrodynamically shaped tufts). The biological content is rich in algae and animals, mainly at the proximity of springs, even ifPhormidium incrustatum is the predominant species. It is associated with several species ofGongrosira, Schizothrix, andOocardium stratum, the latter only known by its specific crystallizations. Among the animals, we point the galleries ofPsychomiidae (Trichoptera= Phrygan) larvae. The algae and animals are associated within a “biological felt” (in the sense ofForel, 1901). Some species are encrusted by calcite crystals of typical habitus (micrite:Phormidium incrustatum, Gongrosira andSchizothrix, ssp; sparite:Oocardium andBatrachospermum), and there are very little diagenetic modifications. The fabric results in an alternation of seasonal light laminations composed of juxtaposed bundles ofPhormidium incrustatum α, and dark laminations due to parallel filaments ofPhormidium incrustatum β. The influence of other algal species on shape and the internal fabric of the laminations is negligible.Phormidium incrustatum tufa are common in Western Europe, and probably have some fossil analogue in the Upper Cretaceous and Tertiary; the strongly differ from most older stromatolitic microstructures. Half of the studied tufa can suffer summer exposure and winter frost but related particular features do not seem to be preserved in the stromatolitic edifices.     

Effect of algal extracts of Phormidium species on growth and development of rice seedlings

Summary The effect of growth-promoting substances in extracts of Phormidium foveolarum, P. tenue, and P. frigidum on rice seedlings has been studied by presoaking seed treatment. The observations show that the growth and development of roots and shoots is markedly stimulated, in seeds treated with P. foveolarum and P. tenue and the results are highly significant statistically.     

Microbially influenced lacustrine carbonates: A comparison of Late Quaternary Lahontan tufa and modern thrombolite from Fayetteville Green Lake,NY

Carbonate microbialites in lakes can serve as valuable indicators of past environments, so long as the biogenicity and depositional setting of the microbialite can be accurately determined. Late Pleistocene to Early Holocene frondose draping tufa deposits from Winnemucca Dry Lake (Nevada, USA), a subbasin of pluvial Lake Lahontan, were examined in outcrop, petrographically, and geochemically to determine whether microbially induced precipitation is a dominant control on deposition. These observations were compared to modern, actively accumulating microbialites from Fayetteville Green Lake (New York, USA) using similar methods. In addition, preserved microbial DNA was extracted from the Lahontan tufa and sequenced to provide a more complete picture of the microbial communities. Tufas are texturally and geochemically similar to modern thrombolitic microbialites from Fayetteville Green Lake, and the stable isotopic composition of organic C, N, inorganic C, and O supports deposition associated with a lacustrine microbial mat environment dominated by photosynthetic processes. DNA extraction and sequencing indicate that photosynthetic microbial builders were present during tufa deposition, primarily Chloroflexi and Proteobacteria with minor abundances of Cyanobacteria and Acidobacteria. Based on the sequencing results, the depositional environment of the tufas can be constrained to the photic zone of the lake, contrasting with some previous interpretations that put tufa formation in deeper waters. Additionally, the presence of a number of mesothermophilic phyla, including Deinococcus–Thermus, indicates that thermal groundwater may have played a role in tufa deposition at sites not previously associated with groundwater influx. The interpretation of frondose tufas as microbially influenced deposits provides new context to interpretations of lake level and past environments in the Lahontan lake basins.     

Effect of algal hormones on stomatal and epidermal development in rice leaves

Effect of presoaking seeds with different concentrations (0.5, 1 and 5%) of algal extracts on the development of stomata and epidermal cells in rice leaves has been investigated. One percent ether extract of Phormidium foveolarum (Mont.) Gomont suspended in water increases the number of stomata and also the total perimeter of stomatal pores, suggesting that the plants are better adopted for photosynthetic activity.     

Effect of inhibitors on calcium carbonate deposition mediated by freshwater algae

Nine green algae, a diatom and three cyanobacteria were shown to precipitate CaCO₃ in batch culture, when grown in the light in a hard water medium containing 68 mg L⁻¹ soluble calcium. The composition of the medium was based on that found in a natural hardwater marina where precipitation of CaCO₃ within algal biofilms occurred. Deposition occurred as a direct result of photosynthesis which caused an increase in the pH of the medium. Once a critical pH had been reached, typically approximately pH 9.0, precipitation began evidenced by a fall in the concentration of soluble calcium in the medium. Certain characteristics of the precipitation process displayed by the diatom Navicula sp. were different to those of the other algae. All algae produced extracellular crystals of irregular morphology. Using a standardized protocol employing the green algae Chlorococcum sp. and Stigeoclonium variabile, the effects of various inhibitors of CaCO₃ nucleation or growth of crystals were studied. Fifteen compounds were screened and assessed for their performance in this context. Most materials effectively delayed deposition of CaCO₃, many decreased precipitation rates and all had a marked effect on crystal morphology. The most effective compound was HEDP (1-hydroxyethylene 1,1 diphosphonic acid), which inhibited precipitation completely at a concentration of 2.5 mg L⁻¹ The use of such compounds to reduce the precipitation of calcium salts within algal biofilms in natural hard waters is discussed.     

Characterization of a Small Cryptic Plasmid,pPFl, from Phormidium foveolarum and Vector Construction

The 1509 bp cryptic plasmid, pPFl, from Phormidium foveolarum, a strain of filamentous cyanobacteria of the LPP group, was completely sequenced. The pPFl nucleotide sequence had 97.8% overall similarity with that of a small plasmid from Plectonema boryanum. The structural organization of pPFl and vector construction are discussed.     

Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates

Heterotrophic CaCO₃-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO₃ precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO₃ was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO₃ precipitation by bacteria.     

Phosphate regime structures species composition in cultured phototrophic biofilms

1. The effect of phosphate on species composition in biofilms was studied under three different phosphate regimes (0.5, 5 and 50 μm ) in two different multi species communities: one composed of the four diatom species Melosira varians, Nitzschia perminuta, Navicula trivialis and Achnanthes lanceolata and one containing these diatom species plus the two cyanobacterial species Leptolyngbya foveolarum and Cylindrospermum stagnale. 2. Algal growth in monocultures and mixtures was measured as chlorophyll a and PAM fluorimetry was applied to document density and physiological condition of the two main groups of photosynthetic organisms in mixed cultures. 3. In phosphate‐replete communities, a single species dominated the community (N. perminuta in the diatom mixture and L. foveolarum in the all species mixture), while in the phosphate‐deprived communities several species persisted, in spite of severe phosphate limitation. 4. We conclude that high supply of phosphate enables the species L. foveolarum, and to a lesser extent N. perminuta, to overgrow biofilm consortia, facilitated by their filamentous growth form, motility or the excretion of inhibitors. The persistence of several species under a low phosphate regime is explained by a less intense interspecific interaction in low‐density biofilms. This clarifies field observations published previously.     

Seasonal variations of pH, pCO2, pO2, HCO3- and Ca2+ in the haemolymph: implications on the calcification physiology in Anodonta cygnea

A study about the relationship between the physical–chemical parameters and the calcium carbonate balance between the haemolymph fluid and mantle calcareous structures was carried out in Anodonta cygnea. An intense peak of HCO₃ ⁻ and a highest pH in December–January months may be understood as a preparation period for creating alkaline conditions. An intense pH decrease from January to February in parallel with the HCO₃ ⁻ reduction seems to indicate the beginning process of carbonate precipitation. On the other hand, the following calcium and HCO₃ ⁻ increases in February–May associated with a continuous and gradual pH fall profile may infer two combined aspects: calcium and HCO₃ ⁻ absorption from external environment and a simultaneous intense calcium carbonate deposition in the haemolymph. So, the pCO₂ peak in this period reflects a subsequent result on equilibrium balance between HCO₃ ⁻ absorption and deposition. The only significant pO₂ increase in the next period, from February to June, is related with an energetic increase to support the metabolic activity favouring the posterior intense pCO₂ peaks. The extended time of CO₂ production in the haemolymph from May to November should induce an increased metabolic acidosis with subsequent intense formation of both HCO₃ ⁻ and Ca²⁺ ions in the same period. This seems to result from CaCO₃ deposits dissolution in the haemolymph, the most direct calcareous source. Additionally, the later increase of metabolic succinic acid during autumn may greatly potentiate this acidosis increasing the dissolution process. Consequently, the pH profile present two simultaneous alkaline peaks in July and October, probably due to a strong HCO₃ ⁻ release from the CaCO₃ dissolution. So, the present seasonal results indicate that in the freshwater bivalve A. cygnea, the low metabolism with higher pH from the early winter is the main cause which may favour a calcareous precipitation, while the high metabolism with lower pH from the early summer may function as an inductor of calcareous dissolution in the haemolymph.     

Bio-deposition of a calcium carbonate layer on degraded limestone by Bacillus species

To obtain a restoring and protective calcite layer on degraded limestone, five different strains of the Bacillus sphaericus group and one strain of Bacillus lentus were tested for their ureolytic driven calcium carbonate precipitation. Although all the Bacillus strains were capable of depositing calcium carbonate, differences occurred in the amount of precipitated calcium carbonate on agar plate colonies. Seven parameters involved in the process were examined: calcite deposition on limestone cubes, pH increase, urea degrading capacity, extracellular polymeric substances (EPS)-production, biofilm formation, ζ-potential and deposition of dense crystal layers. The strain selection for optimal deposition of a dense CaCO₃ layer on limestone, was based on decrease in water absorption rate by treated limestone. Not all of the bacterial strains were effective in the restoration of deteriorated Euville limestone. The best calcite precipitating strains were characterised by high ureolytic efficiency, homogeneous calcite deposition on limestone cubes and a very negative ζ-potential.     

Deposition and cycling of carbon and nitrogen in carbonate mud of the lagoons of Arlington and Sudbury Reefs, Great Barrier Reef

The dynamics of carbon and nitrogen in carbonate mud were examined in the lagoons of Arlington and Sudbury Reefs, Great Barrier Reef. Most (89–93%) of the organic carbon and total nitrogen depositing to the carbonate mud zones was mineralized over a sediment depth of 1 m, with ∼50% of CO₂ produced during microbial decomposition involved in carbonate precipitation/dissolution reactions. There was proportionally little burial of organic carbon (10–11%) or nitrogen (7–10%). Nitrogen budgets suggest rapid turnover of porewater inorganic N pools on the order of hours to a few days. Incubation experiments indicate carbonate dissolution in surface deposits (≤20 cm depth) and carbonate precipitation in deeper sediments. Depth-integrated reaction rates indicate net carbonate precipitation of 7–10 mol CaCO₃ m² year⁻¹ over a depth of 1 m. Budget calculations at the whole-reef scale imply that deposition of CaCO₃ in the mud zones of both lagoons may equate to 50–90% of total reef carbonate production, with organic carbon fluxes equating to nearly all net primary production on each reef. These biogeochemical estimates point to the functional importance of carbonate mud zones in the lagoons of the shelf reefs of the Great Barrier Reef.     

Field detection of urease and carbonic anhydrase activity using rapid and economical tests to assess microbially induced carbonate precipitation

Microbial precipitation of calcium carbonate is a widespread environmental phenomenon that has diverse engineering applications, from building and soil restoration to carbon sequestration. Urease-mediated ureolysis and CO₂ (de)hydration by carbonic anhydrase (CA) are known for their potential to precipitate carbonate minerals, yet many environmental microbial community studies rely on marker gene or metagenomic approaches that are unable to determine in situ activity. Here, we developed fast and cost-effective tests for the field detection of urease and CA activity using pH-sensitive strips inside microcentrifuge tubes that change colour in response to the reaction products of urease (NH₃) and CA (CO₂). The urease assay proved sensitive and useful in the field to detect in situ activity in biofilms from a saline lake, a series of calcareous fens, and ferrous springs, finding relatively high urease activity in lake samples. Incubations of lake microbes with urea resulted in significantly higher CaCO₃ precipitation compared to incubations with a urease inhibitor, showing that the rapid assay indicated an on-site active metabolism potentially mediating carbonate precipitation. The CA assay, however, showed less sensitivity compared to the urease test. While its sensitivity limits its utility, the assay may still be useful as a preliminary indicator given the paucity of other means for detecting CA activity in the field. Field urease, and potentially CA, activity assays complement molecular approaches and facilitate the search for carbonate-precipitating microbes and their in situ activity, which could be applied toward agriculture, engineering and carbon sequestration technologies.     

Involvement of etfA gene during CaCO3 precipitation in Bacillus subtilis biofilm

The eftA gene in Bacillus subtilis has been suggested to be involved in the oxidation/reduction reactions during fatty acid metabolism. Interestingly etfA deletion in B. subtilis results in impairment in CaCO₃ precipitation on the biofilm. Comparisons between the wild type B. subtilis 168 and its etfA mutant during in vitro CaCO₃ crystal precipitation (calcite) revealed changes in phospholipids membrane composition with accumulation of up to 10% of anteiso-C_17:0 and 11% iso-C_17:0 long fatty acids. Ca²⁺ nucleation sites such as dipicolinic acid and teichoic acids seem to contribute to the CaCO₃ precipitation. etfA mutant strain showed up to 40% less dipicolinic acid accumulation compared with B. subtilis 168, while a B. subtilis mutant impaired in teichoic acids synthesis was unable to precipitate CaCO₃. In addition, B. subtilis etfA mutant exhibited acidity production leading to atypical flagella formation and inducing extensive lateral growth on the biofilm when grown on 1.4% agar. From the ecological point of view, this study shows a number of physiological aspects that are involved in CaCO₃ organomineralization on biofilms.     

Extraction, Isolation, and Bioassay of a Gibberellin-like Substance from Phormidium foveolarum

A gibberellin-like substance has been isolated from the blue-greenalga Phormidium foveolarum. This is active in both dwarf pea,dwarf maize, and cucumber hypocotyl bioassays. It is suggestedthat the beneficial effect of pre-soaking seed in an extractof P. foveloarum on growth, yield, and protein content of rice,wheat, and other plants is mainly due to the presence of thissubstance in the alga.     

Engineered applications of ureolytic biomineralization: a review

Microbially-induced calcium carbonate (CaCO₃) precipitation (MICP) is a widely explored and promising technology for use in various engineering applications. In this review, CaCO₃ precipitation induced via urea hydrolysis (ureolysis) is examined for improving construction materials, cementing porous media, hydraulic control, and remediating environmental concerns. The control of MICP is explored through the manipulation of three factors: (1) the ureolytic activity (of microorganisms), (2) the reaction and transport rates of substrates, and (3) the saturation conditions of carbonate minerals. Many combinations of these factors have been researched to spatially and temporally control precipitation. This review discusses how optimization of MICP is attempted for different engineering applications in an effort to highlight the key research and development questions necessary to move MICP technologies toward commercial scale applications.     

New insights into the role of pH and aeration in the bacterial production of calcium carbonate (CaCO3)

Over recent years, the implementation of microbially produced calcium carbonate (CaCO₃) in different industrial and environmental applications has become an alternative for conventional approaches to induce CaCO₃ precipitation. However, there are many factors affecting the biomineralization of CaCO₃, which may restrict its application. In this study, we investigated the effects of pH and aeration as the main two influential parameters on bacterial precipitation of CaCO₃. The results showed that the aeration had a significant effect on bacterial growth and its rise from 0.5 to 4.5 SLPM could produce 4.2 times higher CaCO₃ precipitation. The increase of pH to 12 resulted in 6.3-fold increase in CaCO₃ precipitation as compared to uncontrolled-pH fermentation. Morphological characterization showed that the pH is an effective parameter on CaCO₃ morphology. Calcite was found to be the predominant precipitate during aeration-controlled fermentations, while vaterite was mainly produced at lower pH (up to 10) over controlled-pH fermentations. Further increase in pH resulted in a morphological transition, and vaterite transformed to calcite at the pH ranges between 10 and 12.