Deoxyribonucleic acid base composition of Sphaerotilus natans and Sphaerotilus discophorus.
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Centrifugation↗
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Phylogeny of the Sphaerotilus-Leptothrix group inferred from morphological comparisons, genomic fingerprinting, and 16S ribosomal DNA sequence analyses.
Phase-contrast light microscopy revealed that only one of eight cultivated strains belonging to the Sphaerotilus-Leptothrix group of sheathed bacteria actually produced a sheath in standard growth media. Two Sphaerotilus natans strains produced branched cells, but other morphological characteristics that were used to identify these bacteria were consistent with previously published descriptions. Genomic fingerprints, which were obtained by performing PCR amplification with primers corresponding to enterobacterial repetitive intergenic consensus sequences, were useful for distinguishing between the genera Sphaerotilus and Leptothrix, as well as among individual strains. The complete 16S ribosomal DNA (rDNA) sequences of two strains of "Leptothrix discophora" (strains SP-6 and SS-1) were determined. In addition, partial sequences (approximately 300 nucleotides) of one strain of Leptothrix cholodnii (strain LMG 7171), an unidentified Leptothrix strain (strain NC-1), and four strains of Sphaerotilus natans (strains ATCC 13338T [T = type strain], ATCC 15291, ATCC 29329, and ATCC 29330) were determined. We found that two of the S. natans strains (ATCC 15291 and ATCC 13338T), which differed in morphology and in their genomic fingerprints, had identical sequences in the 300-nucleotide region sequenced. Both parsimony and distance matrix methods were used to infer the evolutionary relationships of the eight strains in a comparison of the 16S rDNA sequences of these organisms with 16S rDNA sequences obtained from ribosomal sequence databases. All of the strains clustered in the Rubrivivax subdivision of the beta subclass of the Proteobacteria, which confirmed previously published conclusions concerning selected individual strains. Additional analyses revealed that all of the S. natans strains clustered in one closely related group, while the Leptothrix strains clustered in two separate lineages that were approximately equidistant from the S. natans cluster. This finding suggests that the tentative species "L. discophora" needs to be more clearly defined and compared with other species belonging to the genus Leptothrix.
Base Sequence↗
Comparative study of different iron compounds in inhibition of sphaerotilus growth.
The effectiveness of iron compounds on growth inhibition of Sphaerotilus species was compared. In this study, two strains of Sphaerotilus were tested with different iron concentrations in a synthetic sewage (S-medium) as formulated by Lackey and Wattie (Sewage Works J. 12:669-684, 1940). For both strains, >80% inhibition of the maximum respiration rate was obtained by the following levels of soluble iron concentrations at pH 6.0: iron citrate, 20 mg/liter as Fe; iron cysteine, 5 mg/liter as Fe; and ferrous sulfate, 10 mg/liter as Fe. At a pH of 6.7 with iron citrate (20 mg/liter as Fe), inhibition of both strains was in excess of 50%. Insoluble iron compounds, such as iron hydroxides and ferrous carbonate, were found to be much less effective than the soluble iron compounds as inhibitors of these two strains. Aged iron hydroxide (500 mg/liter as Fe) produced a 70% inhibition in the maximum respiration rate while fresh iron hydroxide (52 mg/liter as Fe) and ferrous carbonate (500 mg/liter as Fe) produced a 20% inhibition. Chemical analyses of the iron-inhibited Sphaerotilus strains showed a close relationship between the inhibition of the organism's growth and the amount of iron sorbed by the organism.
Journal Article↗
Morphological and biochemical properties of a Sphaerotilus sp. Isolated from paper mill slimes.
Four strains of filamentous bacteria were isolated from slimes collected in different paper mill factories. Morphological and physiological characterization of the isolates indicated an affiliation with the genus Sphaerotilus. However, while the physiological properties of the isolates were almost identical, pronounced physiological differences between the isolates and Sphaerotilus natans DSM 6575(T), DSM 565, and DSM 566 with respect to their ability to metabolize complex polysaccharides, sugars, polyalcohols, or organic acids as carbon sources were detected. In contrast to the analyzed culture collection strains of S. natans, all paper mill isolates were able to grow at elevated temperatures of up to 40 degrees C. Comparative sequence analysis of nearly complete 16S ribosomal DNA (rDNA) sequences from the four new isolates demonstrated that the retrieved sequences were highly similar to each other (99.6 to 99.8% similarity) and to previously published partial 16S rDNA sequences of S. natans DSM 6575(T) and ATCC 15291. Polyphasic characterization of the isolated Sphaerotilus strains revealed interesting adaptations of the strains to the environmental paper mill conditions with regard to temperature tolerance and utilization of cellulose and starch.
Base Sequence↗
Attached growth of Sphaerotilus and mixed populations in a continuous-flow apparatus.
The effects of NH(4)Cl concentration, organic nitrogen compounds, glucose concentration, dissolved oxygen concentration, and flow rate on the attached growth of pure cultures of Sphaerotilus natans and of a mixed population in a continuous-flow apparatus are described. Low concentrations of NH(4)Cl and oxygen, and high flow rates resulted in attached populations that were dominated by Sphaerotilus. The conditions that allowed maximal attached growth in pure culture did not correspond to the conditions that promoted attached growth of Sphaerotilus in a mixed population.
Ammonium Chloride↗
Role of iron deposition in Sphaerotilus discophorus.
Various physiological aspects of the process of iron deposition in Sphaerotilus discophorus were examined to elucidate its role. The values of iron/protein ratios suggested that a direct relationship existed between the iron concentration of the media and the magnitude of final iron deposition. Saturation of the organism's iron deposition system occurred at a 2.0 mM iron concentration, at a value of 0.6 mg of ferric ion per mg of cell protein. Laboratory data indicated that the strain's very low capacity for iron deposition observed at low external iron concentrations makes it unlikely that it is significant in limiting iron in the natural milieu. Under optimal iron concentrations, however, strain SS1 caused precipitation of iron (adsorbed to cellular material) in broth cultures, which was 10 to 100 times that mediated by some "non-iron" microorganisms. The strain's iron requirement, which was found to be between 0.003 and 0.02 mM, is commensurate with that of other microbes. One hundred micrograms of Mn(II) per ml and possibly 10 mug of either Co(II) or Ni(II) per ml could inhibit iron uptake in the deposition system. Sphaerotilus, when tested for its ability to withstand toxic concentrations of certain trace elements (Co, Cu, Mn, Ni, and Cd), demonstrated no exceptional resistance with respect to several other common microorganisms. Final cell yields were not affected by a varying iron concentration for Sphaerotilus growing under conditions of limiting carbon and nitrogen.
Alanine↗
COMPOSITION OF THE SHEATH OF SPHAEROTILUS NATANS.
Romano, Antonio H. (University of Cincinnati, Cincinnati, Ohio) and Joyce P. Peloquin. Composition of the sheath of Sphaerotilus natans. J. Bacteriol. 86:252-258. 1963.-The sheath of Sphaerotilus natans was isolated and subjected to chemical analysis. Isolation of the sheaths was accomplished by incubating cells in the presence of lysozyme and ethylenediaminetetraacetic acid in tris(hydroxymethyl)aminomethane buffer and adding sodium dodecyl sulfate subsequently. Under these conditions, there was complete dissolution of cells. The sheaths, which were left intact by this treatment, were recovered by centrifugation, washed exhaustively, lyophilized, and subjected to analysis. Hydrolysis of the sheath material with 2 n HCl at 100 C resulted in the liberation of reducing sugars amounting to 36% of the dry weight. Amino sugar accounted for 11% of the dry weight. Paper chromatography of hydrolysates showed the presence of glucose and hexosamine. Tests for muramic acid were negative. In addition to carbohydrate, 27% protein and 5.2% lipid were found to be present. Fractionation studies indicated that essentially all of the polysaccharide was associated with a trichloroacetic acid-soluble fraction. The sheath is therefore considered to be a protein-polysaccharide-lipid complex, which is chemically and anatomically distinct from the cell wall and the slime layer. It is hypothesized that this unique structure may be related to the microcapsule found in many gram-negative bacteria, and may represent a structural specialization of this more common structure.
Bacterial Proteins↗
REQUIREMENT OF SPHAEROTILUS FOR CYANOCOBALAMIN.
Okrend, Harold (Rutgers, The State University, New Brunswick, N.J.), and Norman C. Dondero. Requirement of Sphaerotilus for cyanocobalamin. J. Bacteriol. 87:286-292. 1964.-Cultures of Sphaerotilus grew well in a culture medium containing pancreatic digest of casein, glycerol, and yeast autolysate, but grew poorly in a defined medium containing monosodium glutamate, glycerol, and salts. Measurement of dry weight, cell nitrogen, and deoxyribonucleic acid showed that the addition of 1.0 mmug/ml of cyancobalamin to the defined medium gave growth equivalent to that obtained with the pancreatic digest of casein. Methionine, in a concentration of 2 x 10(4)-fold, relieved the requirement for cyanocobalamin, but cobalt did not replace cyanocobalamin nor stimulate growth. Of 38 additional cultures of diverse origin, 19 showed stimulation by visible comparison of growth in tubes; nonquantitative comparisons, however, were not reliable for showing differences in growth of this organism.
Ammonia↗
PATTERN OF SHEATH SYNTHESIS IN SPHAEROTILUS NATANS.
Romano, Antonio H. (University of Cincinnati, Cincinnati, Ohio), and Dorothy J. Geason. Pattern of sheath synthesis in Sphaerotilus natans. J. Bacteriol. 88:1145-1150. 1964.-Formation of the characteristic sheath of Sphaerotilus natans was followed by immunofluorescence. Fluorescent antisheath antibody was obtained by injecting rabbits with purified sheath material from S. natans, fractionating the serum with ammonium sulfate, and conjugating the globulin fraction with fluorescein isothiocyanate. To follow sheath formation, S. natans was grown on slides immersed in Stokes' medium at 28 C for 9 hr, and was reacted with labeled antibody. The slide cultures were then washed to remove unbound antibody, and were incubated further in fresh medium. Samples were removed at intervals and examined by fluorescence microscopy. Old portions of the sheath remained discretely labeled with no diminution in intensity of fluorescence, whereas non-fluorescent new sheath material appeared at the ends of the filaments. These results indicate that sheath synthesis does not take place by intussusception or diffuse intercalation, but by linear extension of pre-existing sheath. This interpretation was confirmed by a reverse procedure, whereby old sheath was reacted with unlabeled antibody, and new sheath was identified by addition of labeled antibody after further incubation. In this procedure, fluorescence was most intense at the growing tips of the filaments.
Animals↗
Isolation and identification of the sudanophilic granules of Sphaerotilus natans.
Rouf, M. A. (Washington State University, Pullman) and J. L. Stokes. Isolation and identification of the sudanophilic granules of Sphaerotilus natans. J. Bacteriol. 83:343-347. 1962.-The characteristic and prominent sudanophilic granules in five strains of Sphaerotilus natans were shown to be composed of poly-beta-hydroxybutyric acid by the method of isolation, melting point determinations, and infrared absorption spectrum analyses. The polymer may account for 11.0 to 22.5% of the dry weight of the cells. It accumulates during active growth of S. natans and decreases sharply in amount after maximal growth. It accumulates to an equal extent in cultures with and without sheaths.
Butyrates↗
[Research on Pb2 + removal from wastewater by Sphaerotilus natans].
As biosorbent used Sphaerotilus natans, influencing factors on removal of Pb2+ were investigated such as ratio of the biomass and lead, pH, temperature and time of adsorption. The results show that Sphaerotilus natans has good effect on adsorption of Pb2+. The process of adsorption reached equilibrium in 10 minutes. Temperature has no significant effect on the adsorption. Removal of Pb2+ approached to 100% under the condition of pH 5.5, 0.6 g/L and c0 < or =20 mg/L. The maximum adsorption amount was 2.1 mmol/g dry biomass . The adsorption was fitted with Freundlich isotherm adsorption equation under the concentration of Pb2+ 0-60 mg/L. HCI and EDTA can desorb the Pb2+ from the bacteria effectively.
Adsorption↗
Selective Growth Inhibition of Sphaerotilus natans and Beggiatoa sp. by Nucleosides.
9-beta-d-Arabinofuranosyladenine (ara-A) has been found to specifically inhibit the growth of Sphaerotilus natans and Beggiatoa sp. at a low concentration (0.78 mug/ml). The nucleoside had no antimicrobial activity against various microorganisms other than Candida albicans at 1,000 mug/ml. 3'-Deoxyadenosine, 2'-deoxyadenosine, formycin, and some derivatives of ara-A also showed inhibitory activity against Sphaerotilus natans. The growth of Beggiatoa sp. was also inhibited by 9-beta-arabinofuranosylhypoxanthine, 3'-deoxyadenosine, 2'-deoxyadenosine, formycin, toyocamycin, tubercidin, and some derivatives of ara-A. ara-A was quite stable in water and had no harmful effect on fish at 200 mug/ml. The possible uses of ara-A and some nucleosides in controlling the proliferation of S. natans and Beggiatoa sp. in the environment are discussed.
Journal Article↗
Growth-promoting activity of spent sulfite liquor for Sphaerotilus natans growing in a continuous-flow apparatus.
The calcium in ammonia-base spent sulfite liquor, unlike that in calcium-base spent liquor, is not available for growth of Sphaerotilus natans. It has not been possible to obtain a mixed slime with Sphaerotilus as the dominant organism in a continuous-flow apparatus fed spent sulfite liquor.
Calcium↗
Measurement of growth and iron deposition in Sphaerotilus discophorus.
Aspects of the physiology of iron deposition of an iron-precipitating strain of Sphaerotilus were investigated in laboratory culture to characterize the process. Measurement of growth (incorporated L-[3H]alanine) and iron deposition (incorporated 59Fe) demonstrated that Sphaerotilus exhibit a characteristic temporal pattern of iron deposition, which is delayed until the latter portion of the exponential or the onset of the stationary growth phase. The growth rate (mu=0.17 h-1) was apparently independent of the iron concentration in the medium. There was, furthermore, no direct correlation between the iron concentration and final cell protein yield. It was concluded from experiments involving growth on artificial substrata (glass cover slips) that sessile populations derived no phsiological advantage (manifested as differences in growth rates) over free-living cells. There was no difference in the rate or onset of iron deposition of attached compared to suspended cells. Blocking of protein synthesis by the addition of chloramphenicol suggested that, once iron deposition was initiated, continued protein synthesis was not required for full expression of this capability. The results suggested that iron deposition may be possibly mediated by certain of the constituents of the organism's sheath.
Alanine↗
Managanese oxidation by Sphaerotilus discophorus.
Johnson, A. H. (Washington State University, Pullman), and J. L. Stokes. Manganese oxidation by Sphaerotilus discophorus. J. Bacteriol. 91:1543-1547. 1966.-Cell suspensions of Sphaerotilus discophorus in 0.02 m potassium phosphate buffer (pH 7.2) readily oxidized MnSO(4) to dark-brown manganic oxide. The latter appeared to be MnO(2), since its color was the same as MnO(2) and, like the latter, it was readily soluble in HCl with the evolution of chlorine. The amount of O(2) consumed in the oxidation of Mn(++) was 63% or more of theoretical. The ability of the cells to oxidize Mn(++) was destroyed by heating the cells at 93 C for 5 min. Only cell suspensions prepared from cultures of S. discophorus grown with MnSO(4) were able to oxidize Mn(++). It was concluded that the oxidation of manganous ions by S. discophorus is catalyzed by an inducible enzyme(s).
Enzymes↗
Ionic strength effect on copper biosorption by Sphaerotilus natans: equilibrium study and dynamic modelling in membrane reactor.
Biosorption of copper by Sphaerotilus natans in different conditions of ionic strength and pH was studied by performing sorption tests in batch and membrane reactors. Equilibrium batch tests evidenced the negative effect of ionic strength and the positive effect of pH on biosorption performances: the highest determined value for copper specific uptake, q, was about 60 mg/g at pH 6 and about 15 mg/g at pH 4. A competitive equilibrium model was successfully fitted to experimental data at different ionic strength levels to account for copper-sodium competition. In membrane reactor tests, experimental profiles of copper concentration in the permeate vs. time did not evidence a significant effect of ionic strength at low pH values (4 and 5). On the other hand a more remarkable effect of ionic strength on copper concentration in the permeate was observed at pH 6. Experimental profiles of continuous biosorption in the membrane reactors were successfully simulated by developing a dynamic model accounting for Cu-Na competition and for binding ability of cells fragments.
Adsorption↗
Paenibacillus hodogayensis sp. nov., capable of degrading the polysaccharide produced by Sphaerotilus natans.
Sphaerotilus natans is a sheathed bacterium often found in activated sludge that has a bulking problem. A bacterial strain that is able to degrade the extracellular polysaccharide produced by S. natans was isolated. The isolate was a spore-forming, aerobic, rod-shaped bacterium. The Gram reaction was variable or negative. The optimum growth temperature was 30 degrees C and the optimum pH was 8. The G+C content of the DNA was 55 mol%. The major cellular fatty acid and respiratory quinone were anteiso-C(15 : 0) and MK-7, respectively. Phylogenetic analysis based on the 16S rRNA gene indicated that the isolate was a member of the genus Paenibacillus. The nearest relative, with a similarity of 94.2 %, was Paenibacillus koleovorans, a bacterium capable of degrading the sheath of S. natans. The phenotypic characteristics of the isolate were apparently different from those of related species in the genus Paenibacillus. It is proposed that the isolate be designated Paenibacillus hodogayensis sp. nov. The type strain is SG(T) (=JCM 12520(T)=KCTC 3919(T)).
Aerobiosis↗
Composition of an extracellular poly-saccharide produced by Sphaerotilus natans.
The capsular polysaccharide of Sphaerotilus natans has been isolated, purified, and analyzed. Chromatographic and chemical analyses performed on acid hydrolyzates of the purified material have shown that the major components are fucose, galactose, glucose, and glucuronic acid in approximately equimolar amounts. Glucose and glucuronic acid are believed to occur as an aldobiuronic acid unit.
Chromatography↗