Bioprecipitation of Calcium Carbonate Crystals by Bacteria Isolated from Saline Environments Grown in Culture Media Amended with Seawater and Real Brine.

The precipitation of calcium carbonate and calcium sulphate by isolated bacteria from seawater and real brine obtained in a desalination plant growth in culture media containing seawater and brine as mineral sources has been studied. However, only bioprecipitation was detected when the bacteria were grown in media with added organic matter. Biomineralization process started rapidly, crystal formation taking place in the beginning a few days after inoculation of media; roughly 90% of total cultivated bacteria showed. Six major colonies with carbonate precipitation capacity dominated bacterial community structure cultivated in heterotrophic platable bacteria medium. Taxonomic identification of these six strains through partial 16S rRNA gene sequences showed their affiliation with Gram-positive Bacillus and Virgibacillus genera. These strains were able to form calcium carbonate minerals, which precipitated as calcite and aragonite crystals and showed bacterial fingerprints or bacteria calcification. Also, carbonic anhydrase activity was observed in three of these isolated bacteria. The results of this research suggest that microbiota isolated from sea water and brine is capable of precipitation of carbonate biominerals, which can occur in situ with mediation of organic matter concentrations. Moreover, calcium carbonate precipitation ability of this microbiota could be of importance in bioremediation of CO2 and calcium in certain environments.

Isolation and metagenomic characterization of bacteria associated with calcium carbonate and struvite precipitation in a pure moving bed biofilm reactor-membrane bioreactor.

A bench-scale pure moving bed bioreactor-membrane bioreactor (MBBR-MBR) used for the treatment of urban wastewater was analyzed for the identification of bacterial strains with the potential capacity for calcium carbonate and struvite biomineral formation. Isolation of mineral-forming strains on calcium carbonate and struvite media revealed six major colonies with a carbonate or struvite precipitation capacity in the biofouling on the membrane surface and showed that heterotrophic bacteria with the ability to precipitate calcium carbonate and struvite constituted ~7.5% of the total platable bacteria. These belonged to the genera Lysinibacillus, Trichococcus, Comamomas and Bacillus. Pyrosequencing analysis of the microbial communities in the suspended cells and membrane biofouling showed a high degree of similarity in all the samples collected with respect to bacterial assemblage. The study of operational taxonomic units (OTUs) identified through pyrosequencing suggested that ~21% of the total bacterial community identified in the biofouling could potentially form calcium carbonate or struvite crystals in the pure MBBR-MBR system used for the treatment of urban wastewater.

Precipitation of minerals by 22 species of moderately halophilic bacteria in artificial marine salts media: influence of salt concentration.

Precipitation of minerals was shown by 22 species of moderately halophilic bacteria in both solid and liquid artificial marine salts media at different concentration and different Mg2+-to-Ca2+ ratio. Precipitation of minerals was observed for all the bacteria used. When salt concentration increased, the quantity and the size of bioliths decreased, the time required for precipitation being increased. The precipitated minerals were calcite, magnesian calcite, aragonite, dolomite, monohydrocalcite, hydromagnesite and struvite in variable proportions, depending on the bacterial species, the salinity and the physical state of the medium; the Mg content of the magnesian calcite also varied according to the same parameters. The precipitated minerals do not correspond exactly to those which could be precipitated inorganically according to the saturation indices. Scanning electron microscopy showed that the formation of the bioliths is initiated by grouping of calcified cells and that the dominant final morphologies were spherulitic with fibrous radiated interiors. It was demonstrated that moderately halophilic bacteria play an active role in the precipitation of carbonates and we hypothesize about this process of biomineralization.

Precipitation of carbonates by Nesterenkonia halobia in liquid media.

We investigated the precipitation of carbonates by Nesterenkonia halobia in a liquid medium at different concentrations of salts and incubation times. N. halobia only produced crystals at salt concentrations of 2.5%, 7.5% and 15%. At 20% salt concentration no crystal formation was observed. Calcite, aragonite and dolomite were precipitated in different quantities, depending on the salinity of the medium and incubation time. Scanning and transmission electron microscopy, microanalysis and electron diffraction were all used to study in detail the morphology, composition and internal structure of the bioliths. We propose a mechanism for biolith formation involving both biological and inorganic processes.

Biomineralization of carbonates by Marinococcus albus and Marinococcus halophilus isolated from the Salar de Atacama (Chile).

We studied the precipitation of carbonates in 17 strains of moderately halophilic, Gram-positive cocci belonging to two species: Marinococcus halophilus and Marinococcus albus, isolated from the Salar de Atacama (Chile). They were cultivated in solid and liquid laboratory media for 42 days at salt concentrations (wt/vol) of 3%, 7.5%, 15%, and 20%. The bioliths precipitated were studied by X-ray diffraction and scanning electron microscopy. M. halophilus formed crystals at each of the salt concentrations, with a maximum number of strains capable of precipitating carbonates at 7.5% and 15% salt concentrations. M. albus did not precipitate at 20% and showed a maximum at 7.5%. This behavior is similar to that of other gram-positive bacteria and differs from that found in gram-negative bacteria. The bioliths precipitated were spherical, generally isolated, with a size of 10-100 microm, varying with salinity. They were of magnesium calcite (CO3 Ca1-x Mgx) with Mg content increasing with increasing salinity and Mg/Ca molar ratio of the culture medium. These results demonstrate the active role played by M. halophilus and M. albus in the precipitation of carbonates.

A study of struvite precipitation and urease activity in bacteria isolated from patients with urinary infections and their possible involvement in the formation of renal calculi.

Struvite precipitation and urease activity were studied in 72 bacterial strains isolated from patients with an urinary infection. The results revealed struvite precipitation by urease-positive, urease-negative and weakly urealitic bacteria. However, in some strains of each of these groups, no precipitation was observed. Variations in pH within the cultures were also investigated as were the mineralogy and morphology of the crystals precipitated using X-ray diffraction and scanning electron microscopy, respectively. The role of urease activity and alkalinization of the medium in struvite precipitation is discussed, as is the possible involvement of bacteria in nucleation processes. We concluded that urease-negative bacteria or those with weak urease activity may also be involved in the formation of struvite renal calculi although urease-positive bacteria seem to make a greater contribution.

Biomineralization of carbonates by Halomonas eurihalina in solid and liquid media with different salinities: crystal formation sequence.

Carbonate precipitation by 20 strains of the moderately halophilic species Halomonas eurihalina in both solid and liquid media was studied. The influence of salinity and temperature on the quantity and type of crystals precipitated was also investigated. Some strains of H. eurihalina formed crystals in all conditions tested. The mineral phases precipitated were magnesium calcite, aragonite and monohydrocalcite in variable proportions depending on various factors such as the type of growth medium employed and its salinity. Scanning electron microscopy and X-ray dispersive energy microanalysis were used to investigate the crystal formation sequence. The process of biolith formation was sequential. It started with chains or filaments of bacteria, giving way to discs which finally produced spherical forms of approximately 50 microns in diameter. We suggest a mechanism of carbonate crystal formation by H. eurihalina.

Precipitation of struvite in urine medium by urease-positive and urease-negative Yersinia strains.

Yersinia strains either urease-positive or urease-negative were examined for precipitation of struvite in human urine at 25 and 37 degrees C. All urease-positive strains and 8 out of 10 urease-negative strains showed the ability to produce these crystals. Incubation time required for precipitation was longer for urease-negative strains and quantity of struvite formed was higher in urease-positive ones. Regarding incubation temperature, no significant influence has been observed.

Formation of bobierrite (magnesium phosphate) crystal aggregates by bacteria from human urine and renal calculi.

The formation of extracellular crystal aggregates of bobierrite [Mg3(PO4)2.8H2O] by bacteria isolated from renal calculi and urine of urolithiasic patients is found with the use of B-17 and B-43 media. The crystal aggregates were observed in the colonies as deposits of brown-yellow coloration, and were identified by X-ray powder diffraction, chemical analysis and scanning electron microscopy. The production of bobierrite by bacteria from human urine and renal calculi implies further versatility of crystal formation by microorganisms, and it may be interesting to investigate the possible relationships between calculi precipitation and urinary infection.

Struvite crystal precipitation by different phenotypes of Yersinia.

Extracellular formation of struvite crystals by Yersinia enterocolitica, Y. frederiksenii, Y. kristensenii and Y intermedia strains was investigated. Precipitation of crystalline structures was found with 19 of the 187 strains tested, its formation being more frequently observed at 25 degrees C than at 37 degrees C. Production of struvite was greater in Y. enterocolitica strains belonging to biotype 1, serogroup 0:7,8 and lysotype Xz, than observed in other phenotypes. Quantitative assay in a liquid medium showed that struvite formation began after 3 d of incubation; production of these crystals increased up to 15 d. Crystalline structures were examined using electron microscopy and their extracellular formation was observed.

[Differentiation of Yersinia enterocolitica 0:3 and 0:9 by precipitation of CaCO3 at 37 degrees C]. / Différenciation de Yersinia enterocolitica 0:3 et 0:9 par la précipitation à 37 degr'es C du CaCO3.

Precipitation of CaCO3 by different Yersinia strains on B-4 agar at 25 and 37 degrees C was studied. The 185 Yersinia strains examined produced crystals at 25 degrees C, but only 121 strains produced crystal at 37 degrees C. Strains of serotype 0:9 produced crystals at 37 degrees C, whereas strains of serotype 0:3 failed to do so. The calcium carbonate precipitation test at 37 degrees C should enable a differentiation between the two serotypes incriminated in human pathology in Europe.