16S rRNA gene-based characterization of bacteria potentially associated with phosphate and carbonate precipitation from a granular autotrophic nitrogen removal bioreactor.

A bench-scale granular autotrophic nitrogen removal bioreactor (completely autotrophic nitrogen removal over nitrite (CANON) system) used for the treatment of synthetic wastewater was analyzed for the identification of microbiota with potential capacity for carbonate and phosphate biomineral formation. 16S ribosomal RNA (rRNA) gene-based studies revealed that different bacterial species found in the granular biomass could trigger the formation of phosphate and calcite minerals in the CANON bioreactor. iTag analysis of the microbial community in the granular biomass with potential ability to precipitate calcium carbonate and hydroxyapatite constituted around 0.79-1.32 % of total bacteria. Specifically, the possible hydroxyapatite-producing Candidatus Accumulibacter had a relative abundance of 0.36-0.38 % and was the highest phosphate-precipitating bacteria in the granular CANON system. With respect to calcite precipitation, the major potential producer was thought to be Stenotrophomonas with a 0.38-0.50 % relative abundance. In conclusion, our study showed evidences that the formation of hydroxyapatite and calcite crystals inside of the granular biomass of a CANON system for the treatment wastewater with high ammonium concentration was a biological process. Therefore, it could be suggested that microorganisms play an important role as a precipitation core and also modified the environment due to their metabolic activities.

Precipitation of phosphate minerals by microorganisms isolated from a fixed-biofilm reactor used for the treatment of domestic wastewater.

The ability of bacteria isolated from a fixed-film bioreactor to precipitate phosphate crystals for the treatment of domestic wastewater in both artificial and natural media was studied. When this was demonstrated in artificial solid media for crystal formation, precipitation took place rapidly, and crystal formation began 3 days after inoculation. The percentage of phosphate-forming bacteria was slightly higher than 75%. Twelve major colonies with phosphate precipitation capacity were the dominant heterotrophic platable bacteria growing aerobically in artificial media. According to their taxonomic affiliations (based on partial sequencing of the 16S rRNA), the 12 strains belonged to the following genera of Gram-negative bacteria: Rhodobacter, Pseudoxanthobacter, Escherichia, Alcaligenes, Roseobacter, Ochrobactrum, Agromyce, Sphingomonas and Paracoccus. The phylogenetic tree shows that most of the identified populations were evolutionarily related to the Alphaproteobacteria (91.66% of sequences). The minerals formed were studied by X-ray diffraction, scanning electron microscopy (SEM), and energy dispersive X-ray microanalysis (EDX). All of these strains formed phosphate crystals and precipitated struvite (MgNH4PO4·6H2O), bobierrite [Mg3(PO4)2·8H2O] and baricite [(MgFe)3(PO4)2·8H2O]. The results obtained in this study show that struvite and spherulite crystals did not show any cell marks. Moreover, phosphate precipitation was observed in the bacterial mass but also near the colonies. Our results suggest that the microbial population contributed to phosphate precipitation by changing the media as a consequence of their metabolic activity. Moreover, the results of this research suggest that bacteria play an active role in the mineral precipitation of soluble phosphate from urban wastewater in submerged fixed-film bioreactors.

Amorphous Ca-phosphate precursors for Ca-carbonate biominerals mediated by Chromohalobacter marismortui.

Although diverse microbial metabolisms are known to induce the precipitation of carbonate minerals, the mechanisms involved in the bacterial mediation, in particular nucleation, are still debated. The study of aragonite precipitation by Chromohalobacter marismortui during the early stages (3-7 days) of culture experiments, and its relation to bacterial metabolic pathways, shows that: (1) carbonate nucleation occurs after precipitation of an amorphous Ca phosphate precursor phase on bacterial cell surfaces and/or embedded in bacterial films; (2) precipitation of this precursor phase results from local high concentrations of PO(4)(3-) and Ca(2+) binding around bacterial cell envelopes; and (3) crystalline nanoparticles, a few hundred nanometres in diametre, form after dissolution of precursor phosphate globules, and later aggregate, allowing the accretion of aragonite bioliths.

Characterisation of Paenibacillus jamilae strains that produce exopolysaccharide during growth on and detoxification of olive mill wastewaters.

A total of 10 bacterial strains were isolated from a compost of corn treated with olive mill wastewaters (OMW) and selected by their capacity to synthesize exopolysaccharides (EPS). Morphological, physiological, biochemical and nutritional tests were used for a phenotypic study. A numerical analysis showed that all strains were 90% similar to each other. A DNA-DNA hybridization assay confirmed that all the strains belonged to Paenibacillus jamilae species. All the characterized strains were able to produce EPS growing on OMW batch cultures. The strain which was able to produce the highest EPS yield was chosen to perform an assay for testing its putative detoxifying activity, and it showed to reduce more than half the toxic capacity of the OMW. The results presented in this study, indicated the possible perspectives for using these bacterial strains to produce EPS and contribute to the bioremediation of the waste waters that are produced in the olive oil elaboration process.

Biomineralization of carbonates by Halobacillus trueperi in solid and liquid media with different salinities.

We investigated the precipitation of carbonates by Halobacillus trueperi in both solid and liquid media at different salt concentrations and different magnesium/calcium ratios. H. trueperi precipitated at all assayed salt concentrations. When salt concentration increased, the quantity and the size of bioliths decreased and the time required increased. The precipitated minerals (determined by X-ray diffraction) were calcite, magnesium calcite and monohydrocalcite in variable proportions depending on the salinity and the physical state of the medium; the magnesium content of the magnesium calcites also varied with regard to the culture type. According to the saturation indices other minerals could also precipitate. Scanning electron microscopy showed that dominant morphologies of the bioliths were spherulitic with fibrous radiated interiors. We show that H. trueperi plays an active role in the precipitation of carbonates and we hypothesize about this process of biomineralization.