Investigation of sporulation in the Desulfotomaculum genus: a genomic comparison with the genera Bacillus and Clostridium.

The genus Desulfotomaculum, belonging to the Firmicutes, comprises strictly anaerobic and endospore-forming bacteria capable of dissimilatory sulfate reduction. These microorganisms are metabolically versatile and are widely distributed in the environment. Spore formation allows them to survive prolonged environmental stress. Information on the mechanism of sporulation in Desulfotomaculum species is scarce. Herein, this process was probed from a genomic standpoint, using the Bacillus subtilis model system as a reference and clostridial sporulation for comparison. Desulfotomaculum falls somewhere in between the Bacillus and Clostridium in terms of conservation of sporulation proteins. Furthermore, it showcased the conservation of a core regulatory cascade throughout genera, while uncovering variability in the initiation of sporulation and the structural characteristics of spores from different genera. In particular, while in Clostridium species sporulation is not initiated by a phosphorelay, Desulfotomaculum species harbour homologues of the B. subtilis proteins involved in this process. Conversely, both Clostridium and Desulfotomaculum species conserve very few B. subtilis structural proteins, particularly those found in the outer layers of the spore. Desulfotomaculum species seem to share greater similarity to the outer layers of Clostridium difficile.

Combined scanning transmission X-ray and electron microscopy for the characterization of bacterial endospores.

Endospores (also referred to as bacterial spores) are bacterial structures formed by several bacterial species of the phylum Firmicutes. Spores form as a response to environmental stress. These structures exhibit remarkable resistance to harsh environmental conditions such as exposure to heat, desiccation, and chemical oxidants. The spores include several layers of protein and peptidoglycan that surround a core harboring DNA as well as high concentrations of calcium and dipicolinic acid (DPA). A combination of scanning transmission X-ray microscopy, scanning transmission electron microscopy, and energy dispersive spectroscopy was used for the direct quantitative characterization of bacterial spores. The concentration and localization of DPA, Ca(2+) , and other elements were determined and compared for the core and cortex of spores from two distinct genera: Bacillus subtilis and Desulfotomaculum reducens. This micro-spectroscopic approach is uniquely suited for the direct study of individual bacterial spores, while classical molecular and biochemical methods access only bulk characteristics.

Sulfur isotope enrichment during maintenance metabolism in the thermophilic sulfate-reducing bacterium Desulfotomaculum putei.

Values of Delta(34)S (=delta(34)S(HS)-delta(34)S(SO(4)), where delta(34)S(HS) and delta(34)S(SO(4)) indicate the differences in the isotopic compositions of the HS(-) and SO(4)(2-) in the eluent, respectively) for many modern marine sediments are in the range of -55 to -75 per thousand, much greater than the -2 to -46 per thousand epsilon(34)S (kinetic isotope enrichment) values commonly observed for microbial sulfate reduction in laboratory batch culture and chemostat experiments. It has been proposed that at extremely low sulfate reduction rates under hypersulfidic conditions with a nonlimited supply of sulfate, isotopic enrichment in laboratory culture experiments should increase to the levels recorded in nature. We examined the effect of extremely low sulfate reduction rates and electron donor limitation on S isotope fractionation by culturing a thermophilic, sulfate-reducing bacterium, Desulfotomaculum putei, in a biomass-recycling culture vessel, or "retentostat." The cell-specific rate of sulfate reduction and the specific growth rate decreased progressively from the exponential phase to the maintenance phase, yielding average maintenance coefficients of 10(-16) to 10(-18) mol of SO(4) cell(-1) h(-1) toward the end of the experiments. Overall S mass and isotopic balance were conserved during the experiment. The differences in the delta(34)S values of the sulfate and sulfide eluting from the retentostat were significantly larger, attaining a maximum Delta(34)S of -20.9 per thousand, than the -9.7 per thousand observed during the batch culture experiment, but differences did not attain the values observed in marine sediments.

Utility of Eucalyptus tereticornis (Smith) bark and Desulfotomaculum nigrificans for the remediation of acid mine drainage.

The efficacy of the bark of Eucalyptus tereticornis (Smith) as an adsorbent for the removal of metal ions and sulphate from acid mine water was assessed. About 96% of Fe, 75% of Zn, 92% of Cu and 41% of sulphate removal was achieved from the acid mine water of pH 2.3 with a concomitant increase in pH value by about two units after interaction with the tree bark, under appropriate conditions. The adsorption isotherms adhered to Freundlich and Langmuir relationships and were exothermic in nature. The free energy of the adsorption process was found to be negative attesting to the feasibility of the reaction. The adsorption kinetics followed the first-order Lagergren rate equation. The filtrate obtained after treatment with E. tereticornis (Sm) bark was found to contain essential elements like potassium, magnesium, calcium, sodium and phosphate apart from carbon which served as a successful growth medium for the sulphate reducing bacteria (SRB) namely Desulfotomaculum nigrificans. Bacterial growth studies showed that about 57% and 72% of sulphate reduction could be achieved at initial pH values of 4.1 and 5.5 respectively of the acid mine water. Pretreatment of the acid mine water with tree bark followed by bioremoval using Dsm. nigrificans resulted in about 75% and 84% respectively of sulphate reduction at pH 4.1 and 5.5, cumulatively by biosorption and bioreduction. The mechanisms of metal ion removal using tree bark and sulphate reduction using Dsm. nigrificans are discussed.

Bioremediation of zinc using Desulfotomaculum nigrificans: bioprecipitation and characterization studies.

Desulfotomaculum nigrificans, a typical sulfate reducing bacterium (SRB) was successfully grown in the presence of 12-210 mg/L of zinc. Complete bioremoval of zinc was achieved in 2 days for 12 mg/L while the bioremoval efficiency was about 70% in 40 days in the presence of 210 mg/L initial concentration of zinc, attesting to the inhibition of bacterial cell growth at higher zinc concentrations. The bioremoval mechanism was predominantly governed by bioprecipitation with biosorption contributing to a minor extent. The amount of protein present in the extracellular secretions obtained by growth of SRB in modified Baars' medium devoid of iron was the highest followed by those obtained in the presence of zinc or iron, in that order. Bioremediation studies carried out using a specially designed set-up, facilitating the transfer of biogenically produced hydrogen sulfide gas to a separate precipitation assembly, confirmed that zinc could be successfully precipitated from its corresponding sulfate solution, varying in concentration from 10 to 20,000 mg/L. Detailed characterization of the various zinc sulfide precipitates by EDAX and X-ray diffraction analysis conformed to wurtzite structure. The isoelectric points of high purity zinc sulfide and that of chemically synthesized, biogenically produced and zinc sulfide precipitated using bacterially produced hydrogen sulfide gas (BPH-ZnS) were located at pH 3, 7.8, 2.8 and 8, respectively.

[Description of Desulfotomaculum nigrificans subsp. salinus as a new species, Desulfotomaculum salinum sp. nov].

This study focused on the physiological, chemotaxonomic, and genotypic characteristics of two thermophilic spore-forming sulfate-reducing bacterial strains, 435T and 781, of which the former has previously been assigned to the subspecies Desulfotomaculum nigrificans subsp. salinus. Both strains reduced sulfate with the resulting production of H2S on media supplemented with H2 + CO2, formate, lactate, pyruvate, malate, fumarate, succinate, methanol, ethanol, propanol, butanol, butyrate, valerate, or palmitate. Lactate oxidation resulted in acetate accumulation; butyrate was oxidized completely, with acetate as an intermediate product. Growth on acetate was slow and weak. Sulfate, sulfite, thiosulfate, and elemental sulfur, but not nitrate, served as electron acceptors for growth with lactate. The bacteria performed dismutation of thiosulfate to sulfate and hydrogen sulfide. In the absence of sulfate, pyruvate but not lactate was fermented. Cytochromes of b and c types were present. The temperature and pH optima for both strains were 60-65 degrees C and pH 7.0. Bacteria grew at 0 to 4.5-6.0% NaCl in the medium, with the optimum being at 0.5-1.0%. Phylogenetic analysis based on a comparison of incomplete 16S rRNA sequences revealed that both strains belonged to the C cluster of the genus Desulfotomaculum, exhibiting 95.5-98.3% homology with the previously described species. The level of DNA-DNA hybridization of strains 435T and 781 with each other was 97%, while that with closely related species D. kuznetsovii 17T was 51-52%. Based on the phenotypic and genotypic properties of strains 435T and 781, it is suggested that they be assigned to a new species: Desulfotomaculum salinum sp. nov., comb. nov. (type strain 435T = VKM B 1492T).