There is a growing demand for silver-based biocides, including both ionic silver forms and metallic nanosilver. The use of metallic nanosilver, typically chemically produced, faces challenges including particle agglomeration, high costs, and upscaling difficulties . Additionally, there exists a need for the development of a more eco-friendly production of nanosilver. In this study, Gram-positive and Gram-negative bacteria were utilized in the non-enzymatic production of silver nanoparticles via the interaction of silver ions and organic compounds present on the bacterial cell. Only lactic acid bacteria, Lactobacillus spp., Pediococcus pentosaceus, Enterococcus faecium, and Lactococcus garvieae, were able to reduce silver. The nanoparticles of the five best producing Lactobacillus spp. were examined more into detail with transmission electron microscopy. Particle localization inside the cell, the mean particle size, and size distribution were species dependent, with Lactobacillus fermentum having the smallest mean particle size of 11.2 nm, the most narrow size distribution, and most nanoparticles associated with the outside of the cells. Furthermore, influence of pH on the reduction process was investigated. With increasing pH, silver recovery increased as well as the reduction rate as indicated by UV-VIS analyses. This study demonstrated that Lactobacillus spp. can be used for a rapid and efficient production of silver nanoparticles.

Microbial reduction of soluble Pd(II) by cells of Shewanella oneidensis MR-1 and of an autoaggregating mutant (COAG) resulted in precipitation of palladium Pd(0) nanoparticles on the cell wall and inside the periplasmic space (bioPd). As a result of biosorption and subsequent bioreduction of Pd(II) with H2, formate, lactate, pyruvate or ethanol as electron donors, recoveries higher than 90% of Pd associated with biomass could be obtained. The bioPd(0) nanoparticles thus obtained had the ability to reductively dehalogenate polychlorinated biphenyl (PCB) congeners in aqueous and sediment matrices. Bioreduction was observed in assays with concentrations up to 1000 mg Pd(II) l(-1) with depletion of soluble Pd(II) of 77.4% and higher. More than 90% decrease of PCB 21 (2,3,4-chloro biphenyl) coupled to formation of its dechlorination products PCB 5 (2,3-chloro biphenyl) and PCB 1 (2-chloro biphenyl) was obtained at a concentration of 1 mg l(-1) within 5 h at 28 degrees C. Bioreductive precipitation of bioPd by S. oneidensis cells mixed with sediment samples contaminated with a mixture of PCB congeners, resulted in dechlorination of both highly and lightly chlorinated PCB congeners adsorbed to the contaminated sediment matrix within 48 h at 28 degrees C. Fifty milligrams per litre of bioPd resulted in a catalytic activity that was comparable to 500 mg l(-1) commercial Pd(0) powder. The high reactivity of 50 mg l(-1) bioPd in the soil suspension was reflected in the reduction of the sum of seven most toxic PCBs to 27% of their initial concentration.

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, zeta-potential and deposition of dense crystal layers. The strain selection for optimal deposition of a dense CaCO(3) 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 zeta-potential.

The interaction between Shewanella oneidensis MR-1 and the soluble metal Pd(II) during the reductive precipitation of Pd(0) determined the size and properties of the precipitated Pd(0) nanoparticles. Assessment of cell viability indicated that the bioreduction of Pd(II) was a detoxification mechanism depending on the Pd(II) concentration and on the presence and properties of the electron donor. The addition of H(2) in the headspace allowed S. oneidensis to resist the toxic effects of Pd(II). Interestingly, 25 mM formate was a less effective electron donor for bioreductive detoxification of Pd(II), since there was a 2 log reduction of culturable cells and a 20% decrease of viable cells within 60 min, followed by a slow recovery. When the ratio of Pd:cell dry weight (CDW) was below 5:2 at a concentration of 50 mg l(-1) Pd(II), most of the cells remained viable. These viable cells precipitated Pd(0) crystals over a relatively larger bacterial surface area and had a particle area that was up to 100 times smaller when compared to Pd(0) crystals formed on non-viable biomass (Pd:CDW ratio of 5:2). The relatively large and densely covering Pd(0) crystals on non-viable biomass exhibited high catalytic reactivity towards hydrophobic molecules such as polychlorinated biphenyls, while the smaller and more dispersed nanocrystals on a viable bacterial carrier exhibited high catalytic reactivity towards the reductive degradation of the anionic pollutant perchlorate.

Trichloroethylene (TCE) is a toxic and recalcitrant groundwater pollutant. An innovative technology using microbial produced Pd(0) nanoparticles for the remediation of TCE contaminated groundwater was developed. The nanoscale bio-Pd particles were precipitated on the biomass of Shewanella oneidensis and hydrogen gas, formate, or formic acid were used as hydrogen donors. Ethane turned out to be the only organic degradation product and no intermediate chlorinated reaction products were detected. Subsequently bio-Pd was implemented in a plate membrane reactor (MR) for the treatment of water containing TCE. In a continuous MR system, containing 50 mg L(-1) bio-Pd, removal rates up to 2,515 mg TCE day(-1) g(-1) Pd were achieved with H(2) gas as hydrogen donor. The measured chloride mass balance confirmed the removal rates. This work shows that a complete, efficient and rapid removal of TCE was achieved with bio-Pd and that a MR system containing bio-Pd and supplied with hydrogen gas offers an alternative for the current remediation technologies of water contaminated with TCE.

The diversity of bacterial groups of activated sludge samples that received wastewater from four different types of industry was investigated by a nested PCR-DGGE (denaturing gradient gel electrophoresis) approach. Specific 16S rRNA primers were chosen for large bacterial groups (Bacteria and alpha-Proteobacteria in particular), which dominate activated sludge communities, as well as for actinomycetes, ammonium oxidisers and methanotrophs (types I and II). In addition primers for the new Acidobacterium kingdom were used to observe their community structure in activated sludge. After this first PCR amplification, a second PCR with bacterial primers yielded 16S rRNA gene fragments that were subsequently separated by DGGE, thus generating 'group-specific DGGE patterns'. The community structure and diversity of the bacterial groups from the different samples was further analysed using different techniques, such as statistical analysis and Shannon diversity index evaluation of the band patterns. By combining the seven DGGE gels, cluster analysis, multidimensional scaling and principal component analysis clearly clustered two of the four activated sludge types separately. It was shown that the combination of molecular and statistical methods can be very useful to differentiate microbial communities.

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In the absence of oxygen, a protective H2 film is formed around an Fe(0) surface, inhibiting the electron flow from this surface. Our study of anoxic corrosion of Fe(0) beads revealed that, in the presence of Shewanella oneidensis MR-1, H2 removal and precipitation of Fe mineral particles on the cell surface are determining processes for corrosion. These two biologically mediated processes were governed by cell density. H2 removal by Shewanella oneidensis was detected at cell concentrations of 1.0 x 10(6) live cells ml-1 and higher and H2 was electron donor for denitrification of NO3-. The removal of the protective H2 layer from Fe(0) beads by Shewanella oneidensis, resulted in an increase of Fe release out of the Fe(0) beads from 153 +/- 25 mg l(-1) to 196 +/- 7 mg l-1 after 20 h. When the cell concentration exceeded 1.0 x 10(8) live cells ml-1, precipitation of iron minerals on the cell surface was characteristic for the greatest percentage of MR-1 cells, whereas micrometre-scale iron precipitates not associated with culturable cell biomass significantly decreased in number. Addition of supernatant of a corrosion assay with high cell concentration induced metabolic activity in a corrosion assay with low cell concentration, resulting in increased H2 consumption and Fe release from Fe(0) beads. Homoserine lactone-like molecules were detected in the supernatant by a bio-assay, suggesting the involvement of a quorum-sensing regulatory mechanism.