The effect of dietary fiber (oat bran) supplement on blood pressure in patients with essential hypertension: A randomized controlled trial.

BACKGROUND AND AIMS: Insufficient dietary fiber (DF) intake is associated with increased blood pressure (BP) and the mode of action is unclear. The intake of DF supplements by participants in previous interventional studies was still far below the amount recommended by the World Health Organization. Therefore, this study aims to explore the effect of supplementing relatively sufficient DF on BP and gut microbiota in patients with essential hypertension (HTN). METHODS AND RESULTS: Fifty participants who met the inclusion criteria were randomly divided into the DF group (n = 25) and control group (n = 25). All the participants received education on regular dietary guidance for HTN. In addition to dietary guidance, one bag of oat bran (30 g/d) supplement (containing DF 8.9 g) was delivered to the DF group. The office BP (oBP), 24 h ambulatory blood pressure, and gut microbiota were measured at baseline and third month. After intervention, the office systolic blood pressure (oSBP; P < 0.001) and office diastolic blood pressure (oDBP; P < 0.028) in the DF group were lower than those in the control group. Similarly, the changes in 24hmaxSBP (P = 0.002), 24hmaxDBP (P = 0.001), 24haveSBP (P < 0.007), and 24haveDBP (P = 0.008) were greater in the DF group than in the control group. The use of antihypertensive drugs in the DF group was significantly reduced (P = 0.021). The ß diversity, including Jaccard (P = 0.008) and Bray-Curtis distance (P = 0.004), showed significant differences (P < 0.05) between two groups by the third month. The changes of Bifidobacterium (P = 0.019) and Spirillum (P = 0.006) in the DF group were significant. CONCLUSIONS: Increased DF (oat bran) supplement improved BP, reduced the amount of antihypertensive drugs, and modulated the gut microbiota. TRIAL REGISTRATION NUMBER: ChiCTR1900024055.

[The functional role of reduced inorganic sulfur compounds in the metabolism of the microaerophilic bacterium Spirillum winogradskii].

Oxidation of reduced sulfur compounds by microaerophilic sulfur bacterium Spirillum winogradskii was found to occur only concomitantly with consumption of an organic substrate and was not linked to their utilization as electron donors in energy metabolism. No enzymes of dissimilatory sulfur metabolism were found in the cells of the sulfur bacterium oxidizing thiosulfate to tetrathionate; oxidation of thiosulfate and sulfide was caused by their reaction with reactive oxygen species (ROS), mostly H2O2 produced in the course of aerobic growth. Decreased lytic effect of ROS in the presence of thiosulfate resulted in a twofold increase in the cell yield under aerobic conditions and more efficient substrate utilization. The latter effect was caused by decreased expense of energy for the biosynthesis of oxygen-protecting polysaccharides. The stimulatory effect of thiosulfate on the growth processes was due to the activation of a number of TCA cycle enzymes producing the intermediates for constructive metabolism, especially of the NADP-dependent malic enzyme. As a result of thiosulfate-induced synthesis of SH-containing cell components, the integral antioxidative activity increased 1.5-fold.

Characterization of bioluminescent derivatives of assimilable organic carbon test bacteria.

The assimilable organic carbon (AOC) test is a standardized measure of the bacterial growth potential of treated water. We describe the design and initial development of an AOC assay that uses bioluminescent derivatives of AOC test bacteria. Our assay is based on the observation that bioluminescence peaks at full cell yield just prior to the onset of the stationary phase during growth in a water sample. Pseudomonas fluorescens P-17 and Spirillum sp. strain NOX bacteria were mutagenized with luxCDABE operon fusion and inducible transposons and were selected on minimal medium. Independent mutants were screened for high luminescence activity and predicted AOC assay sensitivity. All mutants tested were able to grow in tap water under AOC assay conditions. Strains P-17 I5 (with p-aminosalicylate inducer) and NOX I3 were chosen for use in the bioluminescence AOC test. Peak bioluminescence and plate count AOC were linearly related for both test bacteria, though data suggest that the P-17 bioluminescence assay requires more consistent luminescence monitoring. Bioluminescence results were obtained 2 or 3 days postinoculation, compared with 5 days for the ATP luminescence AOC assay and 8 days for the plate count assay. Plate count AOC assay results for nonmutant and bioluminescent bacteria from 36 water samples showed insignificant differences, indicating that the luminescent bacteria retained a full range of AOC measurement capability. This bioluminescence method is amenable to automation with a microplate format with programmable reagent injection.

[Oxidative stress and antioxidant cell protection systems in the microaerophilic bacterium Spirillum winogradskii]. / Okislitel'nyi stress i sistemy antioksidantnoi zashchity kletok u mikroaérofil'nykh bakterii Spirillum winogradskii.

The influence of oxygen availability during cultivation on the biosynthetic processes and enzymatic activities in the microaerophilic bacterium Spirillum winogradskii D-427 was studied, and the roles played by different systems of the defense against oxidation stress were determined. The metabolic adjustments caused by transition from microaerobic (2% O2) aerobic conditions (21% O2 of the gas phase) were found to slow down constructive metabolism and increase synthesis of exopolysaccharides as a means of external protection of cells from excess oxygen. This resulted in a twofold decline of the growth yield coefficient. Even though the low activity of catalase is compensated for by a multifold increase in the activities of other cytoplasmic enzymes protecting from toxic forms of O2--peroxidase and enzymes of the redox system of glutathione (glutathione peroxidase and glutathione reductase)--massive lysis of cells starts in the mid-exponential phase and leads to culture death in the stationary phase because of H2O2 accumulation in the periplasm (up to 10 micrograms/mg protein). The absence in cells of cytochrome-c-peroxidase, a periplasmic enzyme eliminating H2O2, was shown. It follows that the major cause of oxidative stress in cells is that active antioxidant defenses are located in the cytoplasm, whereas H2O2 accumulates in the periplasm due to the lack of cytochrome-c-peroxidase. The addition to the medium of thiosulfate promotes elimination of H2O2, stops cell lysis under aerobic conditions, lends stability to cultures, and results in a threefold increase in the growth yield.

Investigations into the life cycle of the bacterial predator Bdellovibrio bacteriovorus 109J at an interface by atomic force microscopy.

Atomic force microscopy was used to image Bdellovibrio bacteriovorus 109J, a gram-negative bacterial predator that consumes a variety of other gram-negative bacteria. In predator-prey communities grown on filters at hydrated air-solid interfaces, repeated cycles of hunting, invasion, growth, and lysis occurred readily even though the cells were limited to near two-dimensional movement. This system allowed us to image the bacteria directly without extensive preparation or modification, and many of the cells remained alive during imaging. Presented are images of the life cycle in two species of prey organisms, both Escherichia coli (a small prey bacterium that grows two-dimensionally on a surface) and Aquaspirillum serpens (a large prey bacterium that grows three-dimensionally on a surface), including high-resolution images of invaded prey cells called bdelloplasts. We obtained evidence for multiple invasions per prey cell, as well as significant heterogeneity in morphology of bdellovibrios. Mutant host-independent bdellovibrios were observed to have flagella and to excrete a coating that causes the predators to clump together on a surface. Most interestingly, changes in the texture of the cell surface membranes were measured during the course of the invasion cycle. Thus, coupled with our preparation method, atomic force microscopy allowed new observations to be made about Bdellovibrio at an interface. These studies raise important questions about the ways in which bacterial predation at interfaces (air-solid or liquid-solid) may be similar to or different from predation in solution.

Survival potential of Aeromonas hydrophila in freshwaters and nutrient-poor waters in comparison with other bacteria.

The survival of a genetically-marked Aeromonas hydrophila strain was studied in water microcosms using viable counts. Aeromonas hydrophila AWWX1 was shown to survive without decline in viable counts for at least 10 d in three of four filtered-autoclaved freshwaters (surface water and groundwater) and in all examined filtered-autoclaved nutrient-poor waters (bottled spring water, Milli-Q and tap water). However, in the unfiltered waters, a rapid decrease in viable counts of Aer. hydrophila AWWX1 was observed after 1-5 d. The survival of Aer. hydrophila AWWX1 in nutrient-poor waters was compared with that of Pseudomonas fluorescens P17 and Spirillum strain NOX. Survival characteristics were organism- and water-dependent. In the filtered-autoclaved waters, viable counts of Spirillum strain NOX were ca 1 log-unit higher than for Aer. hydrophila AWWX1 and Ps. fluorescens P17. The tested strains Aer. hydrophila AWWX1 and Ps. fluorescens P17 survived 3 to 20, respectively 2 to 4 times better in the filtered-autoclaved waters compared to the unfiltered waters. Apparently, any inherent capability of these micro-organisms to adapt to low-nutrient environments was undone by the presence of the autochthonous microbiota. The present findings that Aer. hydrophila survives very poorly in several drinking waters is of utmost importance towards public health and arises questions about the mechanisms involved.

The relationship between kinetics of substrate-limited transitions and steady-state growth in continuous cultures of Aquaspirillum autotrophicum limited by pyruvate.

Heterotrophic growth at steady state and during transient states caused by the sudden change of the concentration of the limiting factor in the feed medium was investigated experimentally for continuous cultures of Aquaspirillum autotrophicum limited by pyruvate. A model for describing the growth at steady state was selected from three unstructured models after statistical tests of the data. This model postulates that the growth yield increases linearly with the growth rate. Growth during transitions where the substrate remained limiting at all times was fitted with first-order kinetics. Theoretical predictions of these kinetics were derived from the unstructured models used to describe steady state. The predicted rate coefficients of the transients were compared to the experimental coefficients. It appeared that the model which best described steady-state growth also provided the best predictions for growth during the transient state. It is a widespread opinion that unstructured models are adequate to describe growth under steady-state conditions but not to predict transitions in continuous culture. However, for the particular case studied here, no higher degree of complexity was required to describe transitions, provided the growth of the culture was always limited by the substrate.

Investigations on microbial sulfur respiration. Isolation, purification, and characterization of cellular components from Spirillum 5175.

The sulfur-reducing bacterium Spirillum 5175 was investigated with regard to membrane constituents that might be part of the sulfur oxidoreductase which converts elemental sulfur to hydrogen sulfide. Regardless of the electron acceptor used for cultivation of the bacteria, i.e. elemental sulfur, fumarate, or nitrate (Sp. 5175S,F,N), the qualitative pattern of cytochromes and Fe-S proteins did not change significantly, as documented by ultraviolet/visible and electron paramagnetic resonance spectroscopy of oxidized (as isolated) and reduced (dithionite) samples. With elemental sulfur the prominent cytochrome exhibited absorption maxima at 553, 522.5 and 426 nm in the reduced state. In fumarate-grown cells two prominent cytochromes were found with maxima at 561, 551, 530, 521 and 430 nm. Two b-type cytochromes with Em at -198 mV and -20 mV vs the standard hydrogen electrode were identified in the membrane fraction of Sp. 5175F. A yellow pigment was extracted and identified as a flexirubin-type pigment. Although present in large quantities, it seemed not to be involved in the reduction of elemental sulfur. Menaquinone, MK 6 (Mr 580) was the prominent quinone identified in Sp. 5175. Characterization of a second quinone was not attempted because of its much lower concentration. The membrane constituents of Sp. 5175 were solubilized by a variety of detergents and detergent mixtures. A colorimetric procedure with photochemically reduced phenosafranin as the electron donor and cysteamine trisulfide (RS-S-SR, R = -CH2CH2NH2) as the electron acceptor was used to detect sulfur oxidoreductase activity. Three membrane proteins of Sp. 5175 were purified: (1) an [NiFe] hydrogenase, homogeneous by SDS/polyacrylamide gel electrophoresis, with electron paramagnetic resonance signals as isolated at gx,y,z = 2.01, 2.16, 2.33 (100 K), and a strong signal at g = 2.02 below 20 K; (2) a cytochrome b, Fe-S-dependent fumarate reductase, and (3) a protein apparently linked to the sulfur oxidoreductase activity. In contrast to fumarate reductase, no b-type cytochrome was present in the fractions exhibiting sulfur oxidoreductase activity. The presence of Fe-S centers was demonstrated by electron paramagnetic resonance spectroscopy at 10 K. It is not clear whether the c-type cytochrome in the same fractions is part of the sulfur-reducing apparatus of Sp. 5175.

Superviviencia de Azospirillum en rizósfera de Festuca arundinacea / Azospirillum survivial in rhizosphere of Festuca arundinacea

En dos ensayos de inoculación de plántulas de Festuca arundinacea (4 plantas/pote y 20 plantas/pote), con dos cepas de Azospirillum spp. (sp7, ATCC 29145 y cepa G) en condiciones controladas de laboratorio y con dos niveles de iluminación (197 y 274 micronE. m-2.s-1) se estudió la supervivencia de la cepa introducida sp 7 y la evolución del número de microorganismos posibles fijadores de N2 que aparecieron a lo largo del período experimental. Como controles se utilizaron plántulas inoculadas con el mismo número de células autoclavadas. Los resultados muestran que: (i) la densidade de la cepa introducida decrece en ambos ensayos en forma similar observándose a los 22-23 días entre el 4 y el 5% de la densidad inicial, lo que sugiere que las diferentes densidades de plantas e intensidad de iluminación no afectan significativamente la supervivencia del inóculo; (ii) la densidad de dizótrofos posibles totales a los 22-23 días es similar en ambos ensayos. El seguimiento de estas poblaciones realizado en uno de los ensayos sugiere que pueden existir interacciones de distinto tipo con el inóculo y con el inóculo autoclavado, que varían durante el período estudiado. El análisis de los resultados obtenidos destaca el interés de evaluar este tipo de interacciones microbianas durante la ontogenia de la planta

Superviviencia de Azospirillum en rizósfera de Festuca arundinacea / Azospirillum survivial in rhizosphere of Festuca arundinacea

En dos ensayos de inoculación de plántulas de Festuca arundinacea (4 plantas/pote y 20 plantas/pote), con dos cepas de Azospirillum spp. (sp7, ATCC 29145 y cepa G) en condiciones controladas de laboratorio y con dos niveles de iluminación (197 y 274 micronE. m-2.s-1) se estudió la supervivencia de la cepa introducida sp 7 y la evolución del número de microorganismos posibles fijadores de N2 que aparecieron a lo largo del período experimental. Como controles se utilizaron plántulas inoculadas con el mismo número de células autoclavadas. Los resultados muestran que: (i) la densidade de la cepa introducida decrece en ambos ensayos en forma similar observándose a los 22-23 días entre el 4 y el 5% de la densidad inicial, lo que sugiere que las diferentes densidades de plantas e intensidad de iluminación no afectan significativamente la supervivencia del inóculo; (ii) la densidad de dizótrofos posibles totales a los 22-23 días es similar en ambos ensayos. El seguimiento de estas poblaciones realizado en uno de los ensayos sugiere que pueden existir interacciones de distinto tipo con el inóculo y con el inóculo autoclavado, que varían durante el período estudiado. El análisis de los resultados obtenidos destaca el interés de evaluar este tipo de interacciones microbianas durante la ontogenia de la planta (AU)

Components of the regular surface array of Aquaspirillum serpens MW5 and their assembly in vitro.

The two-layered regular surface array of Aquaspirillum serpens MW5 was removed from cell envelopes and dissociated into subunits by treatment with 6 M urea. The surface components reassembled onto an outer membrane surface and self-assembled into planar sheets in vitro in the presence of Ca2+ or Sr2+. The two layers were removed sequentially from cell envelopes by a two-step extraction procedure involving initial treatment with a high-pH buffer to remove the outermost surface layer and subsequent treatment with 6 M urea to remove the innermost layer. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the outer and inner layers of the array were composed of two proteins with molecular weights of 125,000 and 150,000, respectively. The two layers assembled sequentially; the 150,000-molecular-weight protein formed an array on an outer membrane surface, and the 125,000-molecular-weight protein required that array as a template for its in vitro assembly.

On different sensitivities of microorganisms to lowered gravitation.

The influence of lowered gravitation on biomass and CO2 production in B. megaterium, a xerophyte, and Spirillum azotocolligens, an aqueous spirillum, in liquid nutrient medium on a horizontal clinostat at 0.1 g has been studied. As controls we considered: 1) growth under stationary conditions of cultivation with test tubes oriented horizontally; 2) growth on a synchronously revolving centrifuge; and 3) growth on a swing with stirring. A horizontal clinostat at 0.1 g stimulates biomass production and CO2 release in B. megaterium as compared with the controls. Spirillum azotocolligens growth is reduced as a result of clinostating. The best development and CO2 production are observed under stationary conditions. The results do not support the assumption that microorganisms living in water are more resistant to lowered gravitation than those living in soil.

Comparative studies of nitrogen-fixing bacteria associated with grasses in Israel with Azospirillum brasilense.

Free-living, dinitrogen-fixing bacteria associated with roots of grasses were isolated from several locations in Israel. Bacteria with characteristics similar to those of Azospirillum were isolated from Cynodon dactylon roots and were compared with Azospirillum brasilense from Brasil (Sp-7) and from California (Cd). Colonies of the Israeli isolates were yellow and consisted of curved rods, 0.5-0.6 micron in diameter with polar flagella, whereas colonies of A. brasilense were pink (Sp-7) and red (Cd) and the cells were 1.0-1.1 micron in diameter with polar flagella. Ultraviolet absorption spectra of soluble c and membrane-bound c and b cytochromes were similar in all isolates. When grown in semisolid agar medium with or without ammonium chloride all isolates formed a growth zone below the surface. However, they grew best under aerobic conditions in liquid medium containing NH4Cl. All isolates could use salts of malate and lactate, arabinose, and galactose, but not mannitol, as sole carbon sources; they did not need biotin to shorten their lag phase. One Israeli isolate was capable of growing and fixing nitrogen with glucose as a sole carbon source. The Israeli isolates formed aggregates above pH 7.6 in liquid or semisolid medium and were capable of reducing nitrate to nitrogen gas under anaerobic conditions.

Isolation of Azospirullum from diverse geographic regions.

We have isolated Azospirillum (Spirullum lipoferum) from roots of grasses of several genera collected from a number of tropical and subtropical-temperate locations. Pure cultures were obtained from a small percentage of samples; no higher percentage was secured from tropical than from other grasses. Acetylene reduction and distinctive growth in N-free soft agar deeps were inadequate to identify this genus, although helpful in initial screening. Fluorescent antibody tests with antiserum against characterized strains were helpful. There is some evidence that this genus of bacteria may be favored in the rhizoplane.

Light microscopy observations of tetrazolium-reducing bacteria in the endorhizosphere of maize and other grasses in Brazil.

Roots of field-grown tropical maize, Panicum maximum Jacq. and Digitaria decumbens Stent., and of sorghum and wheat grown in monoxenic culture with the diazotroph Spirillum lipoferum (syn. Azospirillum spp.) were examined for tetrazolium-reducing bacteria following incubation of roots in a malate-phosphate buffer-2,3,5-triphenyltetrazolium chloride medium. Bacteria were observed between and in cells of the cortex, in intercellular spaces between the cortex and endodermis, in xylem cells, and in and between pith cells. In maize, colonization of the inner cortex and stele appears to occur in the absence of significant bacterial colonization or collapse of outerlying tissues. Bacteria in the stele remained viable after a 6-h treatment of roots with chloramine-t, indicating that the endodermis was intact. Infection of the inner cortex and stele appears to occur initially in branches, and then to spread longitudinally into main roots. Inter- and intra-cellular infections of the cortex were observed in monoxenic systems. Tetrazolium reduction and prominent crystal formation was not specific for diazotrophic bacteria, but S. lipoferum was isolated from surface-sterilized roots, and S. lipoferum-like organisms were observed in the endorhizosphere. A correlation of inner cortex and stele infections with the presence of branches appears to explain previous observations that excised roots of grasses exhibiting high nitrogenase activity are characteristically branched roots with an intact cortex.

Physiological basis of the selective advantage of a Spirillum sp. in a carbon-limited environment.

A Spirillum sp. and a Pseudomonas sp. possessing crossing substrate saturation curves for L-lactate were isolated from fresh water by chemostat enrichment. Their Ks and mumax values for L-lactate were: Spirillum sp., 23 micrometer and 0.35 h-1, respectively; Pseudomonas sp., 91 micrometer and 0.64 h-1, respectively. Under L-lactate limitation, pseudomonas sp. outgrew Spirillum s. at dilution rates (D) above 0.29 h-1, but the converse occurred at lower D values. The advantage of Spirillum sp. increased with decreasing D until, at D = 0.05 h-1 (i.e. L-lactate concentration of approximately 1 micrometer), Pseudomonas sp. was eliminated from the culture essentially as a non-growing population. In Spirillum sp. the Km for L-lactate transport (5.8 micrometer) was threefold lower than in Pseudomonas sp. (20 micrometer); Spirillum sp. also possessed a higher Vmax for the transport of this substrate. The surface to volume ratio was higher in Spirillum sp. and increased more markedly than in Pseudomonas sp. in response to decreasing D. Thus, a more efficient scavenging capacity contributes to the advantage of Spirillum sp. at low concentrations of the carbon source. Although most of the enzymes of L-lactate catabolism were more active in Pseudomonas sp., NADH oxidase activity was about twice as high in Spirillum sp.; and, unlike Pseudomonas sp., the cytochrome c content of this bacterium increased markedly with decreasing D. A more active and/or more efficient respiratory chain may therefore also play a role in the advantage of Spirillum sp. The other factors which appear to be involved include a lower energy of maintenance of Spirillum sp. [0.016 g L-lactate (g cell dry wt)-1 h-1 compared with 0.066 in Pseudomonas sp.] and a lower minimal growth rate.

Nitrogen fixation, denitrification, and pleomorphic growth in a highly pigmented Spirillum lipoferum.

A strain of Spirillum lipoferum with intense red pigmentation was isolated from the roots of Cynodon dactylon "Coastal." This isolate vigorously reduced acetylene when grown in N-free, Na-malate, semisolid agar, and it was identical to S. lipoferum strain SP7 by standard taxonomic tests. Both S. lipoferum SP7 and the C. dactylon root isolate displayed the unique features of being denitrifiers as well as N2 fixers. The N2-dependent growth curve was biphasic: cells in younger cultures showed the characteristic spiral shape and motility, but those in older cultures developed larger, nonmotile, cystlike forms. Nitrogenase activity seemed associated only with younger spiral forms. The red pigment may be a b- or c-type cytochrome. The strong red color, which this strain develops, could be used as a marker in evaluating soil inoculation experiments.

Reduction of sulfur by spirillum 5175 and syntrophism with Chlorobium.

A small spirillum, designated 5175, was isolated from an anaerobic enrichment culture for Desulfuromonas in which the major medium constituents were acetate and elemental sulfur. The organisms grew only under anaerobic or microaerophilic conditions. Elemental sulfur was formed anaerobically in a malate-sulfide medium, and cell densities of 10(8) cells/ml were obtained. Hydrogen and formate were actively oxidized as substrates for growth under anaerobic conditions; S0, S032-, or S2O32-, but not SO42-, served as electron acceptors and were stoichiometrically reduced to sulfide. Malate or fumarate likewise served as electron acceptors and were reduced to succinate. Nutritional requirements were simple, no vitamins or amino acids being required. For growth in inorganic media when carbon dioxide was the only carbon source, the addition of acetate was required as a source of cell carbon. The organism is gram negative. Cells had a diameter of 0.5 mum and a wavelength of 5.0 mum. Cell suspensions exhibited an absorption spectrum indicative of a cytochrome with peaks in the reduced form at 552, 523, and 416 nm. Well growing syntrophic cultures with Chlorobium were established with formate as the substrate.

Growth of Spirillum lipoferum at constant partial pressures of oxygen, and the properties of its nitrogenase in cell-free extracts.

Spirillum lipoferum, an N2-fixing organism, was grown at constant concentrations of dissolved O2. When supplied with NH4+ aerobically, its doubling time was 1 h; when it fixed N2 microaerophilically, its doubling time was 5-5 to 7 h and the optimal PO2 for growth was 0-005 to 0-007 atm. At its optimal PO2 for growth on N2, S. lipoferum assimilated 8 to 10 mg nitrogen/g carbon substrate used; its efficiency was less at higher PO2 levels. Nitrogenase in cell-free extracts required Mg2+ and Mn2+, and the Fe-protein was activated by Rhodospirillum rubrum activating factor. The nitrogenase had an optimal pH of 7-1 to 7-4 and an apparent Km for acetylene of 0-0036 atm. Extracts of S. lipoferum lost their nitrogenase activity on storage at -18 degrees C, and activity was restored by adding purified Fe-protein from other N2-fixing bacteria.