Effect of a highly concentrated lipopeptide extract of Bacillus subtilis on fungal and bacterial cells.

Lipopeptides produced by Bacillus subtilis are known for their high antifungal activity. The aim of this paper is to show that at high concentration they can damage the surface ultra-structure of bacterial cells. A lipopeptide extract containing iturin and surfactin (5 mg mL(-1)) was prepared after isolation from B. subtilis (strain OG) by solid phase extraction. Analysis by atomic force microscope (AFM) showed that upon evaporation, lipopeptides form large aggregates (0.1-0.2 microm(2)) on the substrates silicon and mica. When the same solution is incubated with fungi and bacteria and the system is allowed to evaporate, dramatic changes are observed on the cells. AFM micrographs show disintegration of the hyphae of Phomopsis phaseoli and the cell walls of Xanthomonas campestris and X. axonopodis. Collapses to fungal and bacterial cells may be a result of formation of pores triggered by micelles and lamellar structures, which are formed above the critical micelar concentration of lipopeptides. As observed for P. phaseoli, the process involves binding, solubilization, and formation of novel structures in which cell wall components are solubilized within lipopeptide vesicles. This is the first report presenting evidences that vesicles of uncharged and negatively charged lipopeptides can alter the morphology of gram-negative bacteria.

Quorum sensing detected by atomic force microscopy imaging of corrals surrounding multicellular arrangement of bacteria.

Connectivity of the glycocalyx covering of small communities of Acidithiobacillus ferrooxidans bacteria deposited on hydrophilic mica plates was imaged by atomic force microscopy. When part of the coverage was removed by water rinsing, an insoluble structure formed by corrals surrounding each individual bacterium was observed. A collective ring structure with clustered bacteria (>or=3) was observed, which indicates that the bacteria perceived the neighborhood in order to grow a protective structure that results in smaller production of exopolysaccharides material. The most surprising aspect of these collective corral structures was that they occur at a low bacterial cell density. The deposited layers were also analyzed by confocal Raman microscopy and shown to contain polysaccharides, protein, and glucoronic acid.

Characterization of nitrogen-fixing cyanobacteria in the Brazilian Amazon floodplain.

The diversity of the free-living nitrogen-fixing cyanobacterial community in the floodplain sediments along the Solimões and Amazon Rivers and some of their tributaries (Japurá, Negro and Madeira) was investigated. Five cyanobacterial genera were morphologically identified, four of which (Nostoc, Calothrix, Cylindrospermum and Fischerella) have not previously been isolated from the Brazilian Amazon floodplain. Nostoc strains were the most commonly found heterocyst-forming cyanobacteria. Five strains (N. muscorum CENA18 and CENA61, N. piscinale CENA21, Cylindrospermum sp. CENA33 and Fischerella sp. CENA19) were selected for growth measurement, ability to fix N2 and phylogenetic analysis, based on their widespread distribution and morphological distinction. Molecular analyses employing 16S rRNA sequences indicated that some of the isolates may represent novel cyanobacterial species. Dinitrogen fixed by these strains was measured indirectly as acetylene reduction activity and ranged from 11.5 to 22.2 nmol C2H4 microg Chl a(-1) h(-1). These results provide evidence of widespread and importance of nitrogen-fixing cyanobacteria as a source of N inputs in the Amazonian ecosystem.