Use of lectins to in situ visualize glycoconjugates of extracellular polymeric substances in acidophilic archaeal biofilms.

Biofilm formation and the production of extracellular polymeric substances (EPS) by meso- and thermoacidophilic metal-oxidizing archaea on relevant substrates have been studied to a limited extent. In order to investigate glycoconjugates, a major part of the EPS, during biofilm formation/bioleaching by archaea on pyrite, a screening with 75 commercially available lectins by fluorescence lectin-binding analysis (FLBA) has been performed. Three representative archaeal species, Ferroplasma acidiphilum DSM 28986, Sulfolobus metallicus DSM 6482(T) and a novel isolate Acidianus sp. DSM 29099 were used. In addition, Acidianus sp. DSM 29099 biofilms on elemental sulfur were studied. The results of FLBA indicate (i) 22 lectins bound to archaeal biofilms on pyrite and 21 lectins were binding to Acidianus sp. DSM 29099 biofilms on elemental sulfur; (ii) major binding patterns, e.g. tightly bound EPS and loosely bound EPS, were detected on both substrates; (iii) the three archaeal species produced various EPS glycoconjugates on pyrite surfaces. Additionally, the substratum induced different EPS glycoconjugates and biofilm structures of cells of Acidianus sp. DSM 29099. Our data provide new insights into interactions between acidophilic archaea on relevant surfaces and also indicate that FLBA is a valuable tool for in situ investigations on archaeal biofilms.

Bioleaching review part A: progress in bioleaching: fundamentals and mechanisms of bacterial metal sulfide oxidation.

Bioleaching of metal sulfides is caused by astonishingly diverse groups of bacteria. Today, at least 11 putative prokaryotic divisions can be related to this phenomenon. In contrast, the dissolution (bio)chemistry of metal sulfides follows only two pathways, which are determined by the acid-solubility of the sulfides: the thiosulfate and the polysulfide pathway. The bacterial cell can effect this sulfide dissolution by "contact" and "non-contact" mechanisms. The non-contact mechanism assumes that the bacteria oxidize only dissolved iron(II) ions to iron(III) ions. The latter can then attack metal sulfides and be reduced to iron(II) ions. The contact mechanism requires attachment of bacteria to the sulfide surface. The primary mechanism for attachment to pyrite is electrostatic in nature. In the case of Acidithiobacillus ferrooxidans, bacterial exopolymers contain iron(III) ions, each complexed by two uronic acid residues. The resulting positive charge allows attachment to the negatively charged pyrite. Thus, the first function of complexed iron(III) ions in the contact mechanism is mediation of cell attachment, while their second function is oxidative dissolution of the metal sulfide, similar to the role of free iron(III) ions in the non-contact mechanism. In both cases, the electrons extracted from the metal sulfide reduce molecular oxygen via a complex redox chain located below the outer membrane, the periplasmic space, and the cytoplasmic membrane of leaching bacteria. The dominance of either At. ferrooxidans or Leptospirillum ferrooxidans in mesophilic leaching habitats is highly likely to result from differences in their biochemical iron(II) oxidation pathways, especially the involvement of rusticyanin.

The EPS of Acidithiobacillus ferrooxidans--a model for structure-function relationships of attached bacteria and their physiology.

To dissolve pyrite or sulphur, leaching bacteria like Acidithiobacillus ferrooxidans attach to these substrata by extracellular polymeric substances (specifically, lipopolysaccharides). The primary attachment to pyrite at pH 2 is mediated by exopolymer-complexed iron(III) ions in an electrostatic interaction with the negatively charged pyrite surface. Cells grown on sulphur exhibit a different composition of the extracellular lipopolysaccharides, namely with increased hydrophobic properties, and do not attach to pyrite. Thus, the cells adapt the chemical composition of their exopolymers to the substrate/substratum. It is concluded that the mechanism of bacterial pyrite oxidation is basically indirect. The actual corrosive agents are iron(III) ions. Preliminary data indicate that active strains complex more iron(III) ions in their EPS than less active ones. Obviously, the exopolymeric layer comprises a reaction space for the regeneration of these ions by the activity of the iron oxidising bacteria.

Large-scale experiments for microbiological evaluation of measures for safeguarding sulfidic mine waste.

In the framework of a German-Romanian scientific cooperation, experiments were performed to evaluate feasible and cheap techniques for the safe storage of mine waste to prevent acid rock drainage (ARD). A large four-chamber percolator (4CP) was installed in a waste heap at Ilba Mine, Romania, to test the effect of biocides and alkaline layers on the bacteria causing acid rock drainage (ARD). The 4CP consisted of four chambers each containing 65 m3 of sulfidic waste material. The 4CP enabled the transfer of laboratory results to a technical scale. The detergent sodiumdodecylsulfate (SDS) was proved to be active against the leaching bacteria. Organotrophic micro-organisms were not effected by the SDS application. The alkaline layers caused an increase of pH, however, a decrease of cell numbers was measured only in adjacent ore layers, but not in the whole ore body. A rapid evaluation of the effects of these countermeasures on ARD formation became possible by microcalorimetric activity measurements for bioleaching.

Bacterial leaching of metal sulfides proceeds by two indirect mechanisms via thiosulfate or via polysulfides and sulfur

The acid-insoluble metal sulfides FeS2, MoS2, and WS2 are chemically attacked by iron(III) hexahydrate ions, generating thiosulfate, which is oxidized to sulfuric acid. Other metal sulfides are attacked by iron(III) ions and by protons, resulting in the formation of elemental sulfur via intermediary polysulfides. Sulfur is biooxidized to sulfuric acid. This explains leaching of metal sulfides by Thiobacillus thiooxidans.

Importance of Extracellular Polymeric Substances from Thiobacillus ferrooxidans for Bioleaching.

Leaching bacteria such as Thiobacillus ferrooxidans attach to pyrite or sulfur by means of extracellular polymeric substances (EPS) (lipopolysaccharides). The primary attachment to pyrite at pH 2 is mediated by exopolymer-complexed iron(III) ions in an electrochemical interaction with the negatively charged pyrite surface. EPS from sulfur cells possess increased hydrophobic properties and do not attach to pyrite, indicating adaptability to the substrate or substratum.

Sulfur chemistry in bacterial leaching of pyrite.

In the case of pyrite bioleaching by Leptospirillum ferrooxidans, an organism without sulfur-oxidizing capacity, besides the production of tetra- and pentathionate, a considerable accumulation of elemental sulfur occurred. A similar result was obtained for chemical oxidation assays with acidic, sterile iron(III) ion-containing solutions. In the case of Thiobacillus ferrooxidans, only slight amounts of elemental sulfur were detectable because of the organism's capacity to oxidize sulfur compounds. In the course of oxidative, chemical pyrite degradation under alkaline conditions, the accumulation of tetrathionate, trithionate, and thiosulfate occurred. The data indicate that thiosulfate, trithionate, tetrathionate, and disulfane-monosulfonic acid are key intermediate sulfur compounds in oxidative pyrite degradation. A novel (cyclic) leaching mechanism is proposed which basically is indirect.

[Occupational rehabilitation of psychiatric patients in the Michaelshoven/Cologne vocational training center--an integrated overall concept]. / Die berufliche Rehabilitation psychisch Behinderter im Berufsförderungswerk Michaelshoven/Köln--Ein integratives Gesamtkonzept.

Vocational retraining centres, sui generis, belong to those vocational rehabilitation facilities where disabled adults are afforded an opportunity for comprehensive occupational reorientation, facilities that operate on the principle of "dual training under one roof", i.e. combine practical and theoretical contents, which documents their close linkage with the mainstream vocational education system practiced in Germany. An essential modification has for some years now been introduced at the Michaelshoven/Cologne vocational retraining centre: the traditional conveyance of skills and knowledge (i.e., acquisition of technical competence) has been replaced by attainment of occupational action competence as the focus of rehabilitative efforts. Hand in hand with this concept rooted in the principle of client-concentration, comes implementation of the principles of integration and holistic approach. Integration means that rehabilitees with mental illness participate in the rehabilitation programmes alongside rehabilitees without mental illness. The holistic approach is expressed in a number of closely coordinated elements ensuring that the specific configuration of persons with mental illness and their way of life are taken into consideration from the very beginning of their stay in the Michaelshoven/Cologne vocational retraining centre. Along with in-depth talks at entry and specific vocational guidance for individuals with mental illness, these elements inter alia include: pre-rehabilitation adjustment, pre-rehabilitation course, an extramural industrial practical, action-oriented training methods throughout, and systematic placement assistance and support--in line with the vocational retraining centre's motto of "picking up our rehabilitees right where they stand".

Microbial diversity in uranium mine waste heaps.

Two different uranium mine waste heaps near Ronneburg, Thuringia, Germany, which contain the remains of the activity of the former uranium-mining Soviet-East German company Wismut AG, were analyzed for the occurrence of lithotrophic and chemoorganotropic leach bacteria. A total of 162 ore samples were taken up to a depth of 5 m. Cell counts of ferrous iron-, sulfur-, sulfur compound-, ammonia-, and nitrite-oxidizing bacteria were determined quantitatively by the most-probable-number technique. Sulfate-, nitrate-, ferric iron-, and manganese-reducing bacteria were also detected. In addition, the metabolic activity of sulfur- and iron-oxidizing bacteria was measured by microcalorimetry. Generally, all microorganisms mentioned above were detectable in the heaps. Aerobic and anaerobic microorganisms thrived up to a depth of 1.5 to 2 m. Up to 99% of Thiobacillus ferrooxidans cells, the dominant leaching bacteria, occurred to this depth. Their numbers correlated with the microbial activity measurements. Samples below 1.5 to 2 m exhibited reduced oxygen concentrations and reduced cell counts for all microorganisms.

Evaluation of Leptospirillum ferrooxidans for Leaching.

The importance of Leptospirillum ferrooxidans for leach processes has been evaluated by studying the lithotrophic flora of three mine biotopes and a heap leaching operation, by percolation experiments with inoculated, sterilized ore, and by morphological, physiological, and genetic investigations of pure and mixed cultures of L. ferrooxidans, Thiobacillus ferrooxidans, and Thiobacillus thiooxidans. In biotopes of 20 degrees C or above, Leptospirillum-like bacteria are as abundant as T. ferrooxidans. Leptospirilli represent at least one-half of the ferrous-iron-oxidizing population. Percolation experiments confirmed this result. Leptospirilli were as numerous as T. ferrooxidans. At reduced temperatures, the generation times of leptospirilli increase more so than those of T. ferrooxidans. At 14 degrees C, Leptospirillum grows slowly and T. ferrooxidans dominates the population. Physiological investigations indicate that L. ferrooxidans is a strict chemolithoautotroph, metabolizing only ferrous iron and pyrite. Even an addition of 0.05% (wt/vol) yeast extract inhibited its growth. The maximum ferrous-iron-oxidizing activity of L. ferrooxidans amounts to about 40% of the activity of T. ferrooxidans. After growth on sulfidic ore, both species exhibit reduced iron-oxidizing activities, L. ferrooxidans exhibiting one-third and T. ferrooxidans exhibiting one-seventh of their maximum activities. Surprisingly, the absolute values are similar. For indirect leaching, L. ferrooxidans is as important as T. ferrooxidans. This was confirmed by the results of percolation experiments. L. ferrooxidans together with T. thiooxidans mobilized metals at least as well as T. ferrooxidans did. The best results were obtained with a mixed culture of all three species.

[Oral iron therapy. Bioavailability and therapeutic effectiveness of ferrous iron in effervescent tablets in posthemorrhagic iron deficiency anemia]. / Orale Eisentherapie. Bioverfügbarkeit und therapeutische Wirksamkeit von Ferro-Eisen als Brausetabletten bei posthämorrhagischer Eisenmangelanämie.

Bio-availability and therapeutic efficacy of two oral ferrous preparations in the form of effervescent tablets (A and A*) were compared. In a randomly controlled trial, postabsorption rise of serum iron was compared intraindividually after oral intake of the effervescent tablets and of an optimally bio-available ferrous ascorbate standard solution (80.5 mg). Afterwards the therapeutic efficacy of both preparations (161 mg daily) was compared with a proprietary iron preparation (B: 150 mg daily) for three months. The trial was conducted on 24 male subjects (aged 20-38 years) who underwent weekly phlebotomies of 500 ml until exhaustion of body iron reserves and development of a mild iron-deficiency anaemia (standard phlebotomy protocol). Relative bioavailability, related to the standard iron solution, was 89% and 104%, respectively, for tablets A and A*. The rise in haemoglobin and ferritin during the three-months treatment was relatively the same for all three preparations: the average daily haemoglobin rise (means +/- SD) was 1.4 +/- 0.5 g/l (A), 1.5 +/- 0.4 g/l (A*) and 1.2 +/- 0.5 g/l (B), respectively, the differences not being statistically significant.

Importance of Hydrogen Sulfide, Thiosulfate, and Methylmercaptan for Growth of Thiobacilli during Simulation of Concrete Corrosion.

Biogenic sulfuric acid corrosion of concrete surfaces caused by thiobacilli was reproduced in simulation experiments. At 9 months after inoculation with thiobacilli, concrete blocks were severely corroded. The sulfur compounds hydrogen sulfide, thiosulfate, and methylmercaptan were tested for their corrosive action. With hydrogen sulfide, severe corrosion was noted. The flora was dominated by Thiobacillus thiooxidans. Thiosulfate led to medium corrosion and a dominance of Thiobacillus neapolitanus and Thiobacillus intermedius. Methylmercaptan resulted in negligible corrosion. A flora of heterotrophs and fungi grew on the blocks. This result implies that methylmercaptan cannot be degraded by thiobacilli.

Water content in convective storm clouds.

The condensed water content of convective storms was measured by the use of a penetrating aircraft. Regions 1 to 2 kilometers in extent and having condensed water contents of about 20 grams per cubic meter were found to be definite features of the cloud interior.