"Use of acidophilic bacteria of the genus Acidithiobacillus to biosynthesize CdS fluorescent nanoparticles (quantum dots) with high tolerance to acidic pH".

The use of bacterial cells to produce fluorescent semiconductor nanoparticles (quantum dots, QDs) represents a green alternative with promising economic potential. In the present work, we report for the first time the biosynthesis of CdS QDs by acidophilic bacteria of the Acidithiobacillus genus. CdS QDs were obtained by exposing A. ferrooxidans, A. thiooxidans and A. caldus cells to sublethal Cd2+ concentrations in the presence of cysteine and glutathione. The fluorescence of cadmium-exposed cells moves from green to red with incubation time, a characteristic property of QDs associated with nanocrystals growth. Biosynthesized nanoparticles (NPs) display an absorption peak at 360nm and a broad emission spectra between 450 and 650nm when excited at 370nm, both characteristic of CdS QDs. Average sizes of 6 and 10nm were determined for green and red NPs, respectively. The importance of cysteine and glutathione on QDs biosynthesis in Acidithiobacillus was related with the generation of H2S. Interestingly, QDs produced by acidophilic bacteria display high tolerance to acidic pH. Absorbance and fluorescence properties of QDs was not affected at pH 2.0, a condition that totally inhibits the fluorescence of QDs produced chemically or biosynthesized by mesophilic bacteria (stable until pH 4.5-5.0). Results presented here constitute the first report of the generation of QDs with improved properties by using extremophile microorganisms.

Use of an acidophilic yeast strain to enable the growth of leaching bacteria on solid media.

In this study, a Candida digboiensis strain was isolated from a heap leaching plant in Zambia and used in double-layer agar plate to efficiently isolate and purify leaching bacteria. Unlike Acidiphilium sp., the yeast strain was tetrathionate tolerant and could metabolize a great range of organic compounds including organic acids. These properties allowed the yeast strain to enable and fasten the growth of iron and sulfur oxidizers on double-layer agar plate. The isolates were identified as Acidithiobacillus ferrooxidans FOX1, Leptospirillun ferriphilum BN, and Acidithiobacillus thiooxidans ZMB. These three leaching bacteria were inhibited by organic acids such as acetic and propionic acids; however, their activities were enhanced by Candida digboiensis NB under dissolved organic matter stress.

Comparison and evaluation of immobilization methods for preparing bacterial probes using acidophilic bioleaching bacteria Acidithiobacillus thiooxidans for AFM studies.

We evaluated different strategies for constructing bacterial probes for atomic force microscopy studies of bioleaching Acidithiobacillus thiooxidans interacting with pyrite mineral surfaces. Of three available techniques, the bacterial colloidal probe technique is the most reliable and provides a versatile platform for quantifying true interactive forces between bioleaching microorganisms and mineral surfaces.

Adaptive changes of chemolithoautotrophic acidophilic sulfur-oxidizing bacteria during growth in sewage sludge.

A chemolithoauthotrophic, acidophilic, sulfur-oxidizing strain was isolated from sewage sludge and identified as Acidithiobacillus thiooxidans. The morphology and physiology of the isolate grown in mineral medium or sterilized sewage sludge were investigated. Morphological and ultrastructural differences between cells grown in mineral medium and sewage sludge were clearly visible. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed some changes in the protein expression profiles in the periplasmic fraction as well as a lower level of cytochromes. Adaptation of A. thiooxidans to sewage sludge was not only a physiological process but also included genetic changes. Restriction fragment length polymorphism analysis using pulsed field gel electrophoresis showed structural changes in chromosomal DNA of such bacteria. Most of the restriction fragments were highly conserved and shared by strains grown under different conditions. Cultivation in mineral medium did, however, lead to the appearance of an additional restriction fragment. In combination, the obtained results provide evidence of adaptive responses by A. thioxidans during growth in sewage sludge and confirm that this bacteria can be useful in biotechnologies of heavy metal bioleaching from different environments polluted with hazardous compounds.

[Morphogenetic features of Sulfobacillus thermosulfidooxidans]. / Osobennosti morfogeneza Sulfobacillus thermosulfidooxidans.

The morphogenesis of Sulfobacillus thermosulfidooxidans BKM B-1269 was studied by electron microscopy at the vegetative stage of growth on a medium with pyrite as the source of energy. Young growing cultures of this organism were found to be very polymorphous: along with rod-like cells of various dimensions (0.6--0.8x1.0--3.0 mcm) which were predominant, coccoid, pyriform, clavate and dumbbell-like cells were also encountered. Certain cells have a tendency for branching. The cells can divide by sudden breaking of the parent cell along a partition being formed. These breaks are often incomplete, thus giving rise to cellular aggregates in the shape of palisades, bent chains, rings and Y-formations. The above characteristics of S. thermosulfidooxidans morphogenesis suggest that the genus Sulfobacillus is related to Corynebacterium-like organisms and actinomycetes. However, this genus differs in its morphology from bacilli though both (as well as actinomycetes) are capable of spore formation.

Acidostability of speroplasts prepared from Thiobacillus thiooxidans.

Thiobacillus thiooxidans was acidostable even in the absence of its respiratory substrate, elementary sulfur. This suggests that the acidostability of the bacterium was enery-independent. The organism was subjected to osmotic shock with 0.75 M sucrose at 0 degrees C and then treated with snail intestinal juice in the presence of 0.3 M sucrose. The decrease in the optical density of the sample thus prepared on dilution with deionized water and electron microscopic observation of the sample showed that spheroplasts were formed from the bacterium by this procedure. Spheroplasts were able to respire sulfur and their respiratory activity was acidostable. Spheroplasts, when treated with Nagase, proteolytic enzyme, lost their acidostability, and some protein components disappeared from the membrane fraction. This suggests that the acidostability of the bacterium may be related to protein conponents of the membrane.

[Function of surface membrane structures in Thiobacillus thiooxidans]. / O funktsii poverkhnostnykh membrannykh struktur Thiobacillus thiooxidans

The function of the surface membrane structures was studied with cytochemical techniques on ultrathin sections of Thiobacillus thiooxidans. The transport of elementary sulphur inside the cell involves the surface membrane structures, while oxidation of the sulphur to sulphuric acid takes place on the outer surface of the cytoplasmic membrane. The surface membrane structures are supposed also to participate in the primary dissolution of elementary sulphur at the site of contact of the cells with the mineral.