Metabolism of daidzein by intestinal bacteria from rhesus monkeys (Macaca mulatta)

PURPOSE: To identify the metabolites produced from an isoflavonoid, daidzein, by colonic bacteria of rhesus monkeys. METHODS: The metabolism of daidzein by the fecal bacteria of nine monkeys was investigated. Daidzein was incubated anaerobically with fecal bacteria, and the metabolites were analyzed by use of liquid chromatography and mass spectrometry. RESULTS: The fecal bacteria of all of the monkeys metabolized daidzein to various extents. Dihydrodaidzein was found in cultures of fecal bacteria from two monkeys; dihydrodaidzein and equol were found in cultures from four monkeys; dihydrodaidzein, equol, and an unknown metabolite (MW = 244) were found in cultures from one monkey; and dihydrodaidzein and the unknown metabolite were found in cultures from two monkeys. CONCLUSIONS: Similar to that in humans, variation was evident in the metabolism of isoflavonoids by fecal bacteria from rhesus monkeys. Some metabolites produced by fecal bacteria from monkeys were the same as those produced by fecal bacteria from humans.     

The influence of bacteria on the arsenic species produced by laboratory cultures of the marine phytoplankton Dunaliella tertiolecta

To assess whether bacteria influence the biogeochemical cycling of arsenic by laboratory cultures of the marine phytoplankton Dunaliella tertiolecta, the arsenic species produced by D. tertiolecta were compared in “operationally sterile” and bacteria spiked cultures. It was observed that glycerol (Gly-) arsenoriboside (41–78 %), phosphate (PO₄?) arsenoriboside (7–38 %) and arsenate (As(V)) (15–21 %) were the major water-soluble arsenic species in all D. tertiolecta cultures irrespective of whether cultures were operationally sterile or contained added bacteria. PO₄-riboside (46–74 %) and Gly-riboside (24–36 %) were also the major arsenic species in hydrolysed lipid extracts of D. tertiolecta irrespective of whether cultures were operationally sterile or contained bacteria. In addition to similarities in the arsenic species produced, total arsenic concentrations and culture growth did not differ relative to whether cultures were operationally sterile or not. Similar bacterial strains were identified in all D. tertiolecta cultures irrespective of whether bacteria were added or not. Consequently, it is evident that the presence of “foreign” or “added” bacteria in D. tertiolecta has minimal influence on the metabolism and cycling of arsenic by phytoplankton. Thus, the use of laboratory phytoplankton cultures containing bacteria may be appropriate means to investigate arsenic biogeochemical cycling unlike previously believed.     

Pressure measurement to evaluate ethanol or lactic acid production during glucose fermentation by yeast or heterofermentative bacteria in pure and mixed culture

A rapid and simple technique to follow CO2 release during fermentation of glucose by heterofermentative bacteria or yeasts was used in order to evaluate ethanol and lactate production in pure and mixed cultures of yeast and bacteria. In pure cultures, good correlations were found between gas pressure variations (deltaP) and ethanol or lactate production by yeasts or heterofermentative bacteria, and ratios between deltaP and ethanol or lactate produced could be established. In mixed cultures, ratios between maximal deltaP and total amount of glucose consumed were determined. It was thus possible to evaluate the amount of glucose that was consumed by each strain and then deduce the bacterial lactate production. Good results were obtained for mixed cultures of yeast and homofermentative bacteria. This technique may be useful to evaluate the activity of strains in mixed cultures of yeast and lactic acid bacteria.     

Decontamination of bacterial infection of monolayer cultures with a specific bacteriophage

Summary A few cell lines and primary monolayer cultures were accidentally infected by bacteria. These cultures were successfully decontaminated by means of the specific bacteriophage virus after quick identification of the responsible bacteria. This method presents a practical interest for preservation of valuable cultures. This work is supported by the Institut National de la Sante et de la Recherche Medicale (France) and the Fondation pour la Recherche Medicale (France).     

Epiphytic bacteria on the Antarctic ice diatom Amphiprora kufferathii Manguin cleave hydrogen peroxide produced during algal photosynthesis

The Antarctic ice diatom Amphiprora kufferathii Manguin is always accompanied by epiphytic bacteria in its natural habitat. To investigate the nature of this relationship, axenic cultures of A. kufferathii were obtained by ampicillin treatment. Diatom cultures without bacteria were less dense. The bacteria were shown to consume hydrogen peroxide produced by the diatom during photosysnthesis and algal photosynthesis after a hydrogen peroxide shock recovered faster in the presence of bacteria. Three proteobacterial strains isolated from a culture of A. kufferathii were phylogenetically affiliated with the alphaproteobacterial genus Sulfitobacter, the gammaproteobacterial genus Colwellia, and the genus Pibocella of the Bacteriodetes. Native protein gel electrophoresis and enzyme activity staining revealed the presence of superoxide dismutase and glutathione reductase in the isolated bacteria and in A. kufferathii cultures. Catalase was detected in bacterial extracts but not in axenic cultures of A. kufferathii. These observations indicate that the epiphytic bacteria make a significant contribution to the diatom's antioxidative defences. The relationship between the bacteria and A. kufferathii seems to be beneficial for both partners and enhances growth of Amphiprora in the sea ice.     

Carbohydrate specificity of lectins from luminescent bacteria

The presence of lectins on a cell surface was demonstrated for 70 cultures of luminescent bacteria using hemagglutination reactions. It was shown that hemagglutination of luminescent bacteria is inhibited by glucose, maltose, fructose, mannose, and N-acetyl-D-glucosamine. The differences in the inhibition of hemagglutination of luminescent and nonluminescent (spontaneous mutants) symbiotic cultures by N-acetyl-D-galactosamine were revealed. The fact that N-acetyl-D-galactosamine inhibits hemagglutination of the luminescent symbiotic bacteria but does not inhibit hemagglutination of the symbiotic cultures lacking luminescence suggests that lectins with N-acetyl-D-galactosamine specificity are possibly involved in the formation and functioning of the symbiosis of luminescent bacteria with marine animals possessing luminous organs.     

Response of populations of human faecal bacteria to viscosity in vitro

The effects of viscosity on cultures of human faecal bacteria in vitro were studied. The composition of the microbial populations which developed during in vitro incubation were affected by viscosity. The proportion of Bacteroides species was decreased at high viscosities and it is concluded that the 'viscotolerance' of a bacterial population contributes towards its composition. Viscosity alters the enzyme activity of cultures. Cell-associated total, and specific, beta-galactosidase activities of cultures of human faecal bacteria were decreased at high viscosity.     

Bacterial influence on the production of paralytic shellfish toxins by dinoflagellated algae

This study investigated the role of intracellular and extracellular bacteria in the production ofparalytic shellfish toxins by dinoflagellated algal cells. Three strains of the toxic dinoflagellatespecies, Alexandrium tamarense , were purified by external bacteria using penicillin G (Pen.G) at levels of 500 and 1000 p.p.m. Levels of toxicity of the resulting purified dinoflagellatecultures were similar to those of the original strains contaminated with external bacteria,indicating that the external bacteria had no influence on toxicity. No bacterial colony formingunits (cfu) arose from disruption of algal cells derived from penicillin-treated cultures, indicatingthat intracellular bacteria were not responsible for the toxicity of cultures.     

Response of populations of human faecal bacteria to viscosity in vitro

The effects of viscosity on cultures of human faecal bacteria in vitro were studied. The composition of the microbial populations which developed during in vitro incubation were affected by viscosity. The proportion of Bacteroides species was decreased at high viscosities and it is concluded that the 'viscotolerance' of a bacterial population contributes towards its composition. Viscosity alters the enzyme activity of cultures. Cell-associated total, and specific, β-galactosidase activities of cultures of human faecal bacteria were decreased at high viscosity.     

Selective outgrowth of fimbriate bacteria in static liquid medium

Competitive mixed cultures were grown from inocula of a large number of bacteria of a genotypically nonfimbriate (fim(-)) strain of Salmonella typhimurium and a small number of a genotypically fimbriate (fim(+)) variant strain that formed type 1 fimbriae and had been derived from the fim(-) strain by phage transduction. The fim(+) strain differed from the fim(-) strain in fermenting l-rhamnose (rha(+)), and the viable fim(+) and fim(-) bacteria present in the cultures after different periods at 37 C were counted differentially in platings on rhamnose media. When the cultures were grown under aerobic static conditions in tubes of nutrient broth, the fim(+) bacteria rapidly outgrew the fim(-) bacteria, so that, although starting as a small minority (e.g., 1 in 10(7)), they approached or surpassed the number of the fim(-) in 48 hr. A pellicle consisting of fimbriate bacteria was formed on the surface of the broth between 6 and 24 hr, and it is thought that the advantage of access to atmospheric oxygen enjoyed by these bacteria in the pellicle enabled them to outgrow the fim(-) bacteria confined in the oxygen-depleted broth. The fim(+) bacteria did not show selective outgrowth in mixed cultures grown in broth aerated by continuous shaking, in static broth incubated anaerobically in hydrogen, and on aerobic agar plates, i.e., under conditions not allowing an advantage from pellicle formation. The outgrowth of fim(+) bacteria in aerobic static broth was prevented by the addition of alpha-methylmannoside, a substance that inhibits the adhesive and early pellicle-forming properties of bacteria with type 1 fimbriae. A motile flagellate (fla(+)) variant of a fim(-)fla(-) strain of S. typhimurium outgrew its parent strain in mixed cultures in aerobic static broth, but the selective advantage conferred by motility was weaker than that conferred by fimbriation.     

Stimulation of Lactic Acid Bacteria by a Micrococcus Isolate: Evidence for Multiple Effects

Growth of, and rate of acid production by, six cultures of lactic acid bacteria were increased in the presence of Micrococcus isolate F4 or a preparation of its capsular material. Concentrations of hydrogen peroxide found in pure cultures of the lactic acid bacteria were not detectable, or were greatly reduced, in mixed culture with Micrococcus isolate F4. The capsular material was not as effective as whole cells in preventing accumulation of H(2)O(2). Catalase stimulated growth of, and the rate of acid production by, the lactic acid bacteria, but not to the same extent as Micrococcus isolate F4 in some cultures. The existence of two mechanisms for micrococcal stimulation of the lactic acid bacteria is postulated. One mechanism involves removal of H(2)O(2); the other has not been characterized.     

Antibiosis plays a role in the context of direct interaction during antagonism of Paenibacillus polymyxa towards Fusarium oxysporum

Interaction of Fusarium oxysporum and Paenibacillus polymyxa starts with polar attachment of bacteria to the fungal hyphae followed by the formation of a large cluster of non-motile cells embedded in an extracellular matrix in which the bacteria develop endospores. Enumeration of fungal viable counts showed that less than one of 36,000 colony-forming units survived in paired cultures for 71 h. Effective antagonism was not observed below pH5 and was specific for the bacterial species. Development of F. oxysporum was inhibited in cell-free filtrates derived from cultures of P. polymyxa, but was much more strongly repressed in the presence of living bacteria. Furthermore, recovery of fungal growth started immediately after addition of antibiotics to paired cultures. Restoration of fungal growth was enhanced in filtrates that were supplemented with MgCl2, which suggests that anti-fungal compounds produced by the bacteria were counteracted by magnesium ions. In paired cultures, fungal counts remained very low, even in the presence of the magnesium salt. This study clearly showed that P. polymyxa antagonizes the plant pathogenic fungus F. oxysporum in liquid medium by means of an interaction process in which the presence of living bacteria is a prerequisite for continuous suppression of fungal growth.     

Preservation of microorganisms

Cultures of bacteria, yeasts and fungi were grown on common agar media in normal culture tubes. About 1 week after maximum growth the cotton plugs of the tubes were replaced by sterile rubber seals. The cultures were stored in the dark at room temperature. Tests for viability of cultures were made after periods between 1 and 10 years. The results of this simple method show long survival periods of many bacteria and yeasts.     

Effect of long-term lead exposure on the seawater and sediment bacteria from heterogeneous continuous flow cultures

Lead-influenced changes of the composition of seawater and sediment bacteria were studied in two flow cultures run with lead-contaminated artificial seawater (1 mg Pb²⁺1^–1) and one control culture. During the experiment viable counts of physiological groups of bacteria from the control culture were not significantly different from that of the lead-contaminated cultures. Lead tolerance of seawater and sediment bacteria strains was investigated. Comparisons of growth yields showed that lead tolerance of seawater and sediment bacteria was lost again if the bacteria were cultivated in a medium without lead. Lead tolerance could not be demonstrated for the sediment bacteria of one lead-contaminated culture. Heterotrophic uptake measurements with radioactive glucose indicated that seawater bacteria from the lead-contaminated cultures became adapted to lead pollution. The sediment bacteria, however, did not reveal lead tolerance by this method. Fluctuations in lead content of the sediment as well as of the overlying seawater gave indications of adsorption-desorption processes between seawater and sediment. Lead was not homogeneously distributed at the sediment surface.     

Degradation of Putrescine and Cadaverine in Seawater Cultures by Marine Bacteria

Marine bacteria removed two diamines, putrescine and cadaverine, from coastal seawater supplemented only with these compounds. Batch cultures of natural bacterial communities were grown in filtered seawater (0.05 μm) supplemented with 500 μg of putrescine or cadaverine per liter. Increases in bacterial cell number were counted with an epifluorescence microscope after acridine orange staining. Removal of diamines from seawater was monitored by high-performance liquid chromatography. Diamines were removed from the seawater cultures within 48 h with no corresponding increase in bacterial yield, growth rate, or viability relative to control (unsupplemented) cultures. Shipboard experiments with open-ocean deep water (1,500 m) showed similar, if slower, removal of putrescine from seawater. Unlike uptake experiments with amino acids, labeled putrescine experiments indicated that most putrescine carbon is mineralized to CO₂ rather than assimilated by the bacteria. After growth in unsupplemented control cultures, the bacteria showed a significant potential to mineralize putrescine, indicating a general degradation potential for this compound by marine bacteria even if the compound was not present during growth. Indicators of metabolic activity such as glucose and glutamic acid uptake and mineralization were not affected by the presence of putrescine. This shows that at the concentrations added, the diamines are not toxic, and therefore detoxification was not the reason for degradation of the diamines by the bacteria.     

Bacteriocinogenic properties of Proteus]

As revealed, bacteriocinogenia was widespread in bacteria of the Proteus genus (85 +/- 3.87%). Proteus cultures isolated from various sources failed to differ by the number of bacteriocinogenic cultures and by bacteriocinogenic activity. Bacteriocinogenic cultures were mostly revealed among the Proteius mirabilis strains.     

Lactic acid bacteria associated with wine grapes from several Australian vineyards

AIMS: The detection and isolation of lactic acid bacteria by enrichment methods from wine grapes cultivated in vineyards located in New South Wales, Australia. METHODS AND RESULTS: Enrichment cultures in de Man, Rogosa and Sharpe (MRS) broth, MRS + ethanol (5%), MRS broth supplemented with 15% (v/v) tomato juice (MRST), pH 5.5 and 3.5 and autoenrichment in grape juice homogenate were used to detect lactic acid bacteria on wine grapes. Bacteria were isolated from enrichment cultures by plating onto MRS and MRST agar and identified by 16S rDNA sequence analysis and phenotypical methods. A molecular method, PCR-denaturing gradient gel electrophoresis (DGGE) was also used to examine the bacteria that developed in enrichment cultures. Species of Lactobacillus, Enterococcus, Lactococcus and Weissella were detected in enrichments by plating and PCR-DGGE. Other bacteria (Sporolactobacillus, Asaia, Bacillus ssp.) were also found in some enrichment cultures. The principal malolactic bacterium, Oenococcus oeni, was not isolated. CONCLUSIONS: The incidence and populations of lactic acid bacteria on wine grapes were very low. Damaged grape berries showed a greater presence of these bacteria than undamaged berries. The diversity of bacterial species isolated from the grapes was greater than those previously reported and represented both lactic acid bacteria and nonlactic acid bacteria. Some of these bacteria (i.e. Lactobacillus lindneri, Lactobacillus kunkeei) could be detrimental to wine production. Oenococcus oeni was not found on grapes, but its recovery could be obscured by overgrowth from other species. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactic acid bacteria are significant in wine production because they conduct the malolactic fermentation and cause stuck or sluggish alcoholic fermentation and wine spoilage. This study investigates wine grapes as a potential source of these bacteria.     

Microbial Utilization of Pinus radiata Bark

A screening program using suspensions of ground bark in mineral salts media, or extracts prepared from ground bark by treating with hot water, sulfuric acid, ammonium hydroxide, or sodium hydroxide, yielded more than 200 pure cultures of fungi, yeasts, and bacteria. Only 38 of these have good growth on liquid bark media. All were filamentous fungi, although many bacteria and yeasts were among the cultures that failed to give appreciable growth. Species of Penicillium, Scopulariopsis, Aspergillus, Trichoderma, Cladosporium, and Fusarium were among the most actively growing cultures. Cell biomass yields, as measured by cell nitrogen determination, were too low for economic production of single cell protein.     

Formation of N-Carboxymethyl Amino Acid from the Reaction of α-Amino Acid with Glyoxal

Enrichment cultures in a medium containing 0.1% methanol and 0.1% bicarbonate at pH 7.0 under anaerobic conditions in the light became mainly green in color. Forty-four enrichment cultures, which showed abundant growth, were obtained from 46 different sources and found to contain cells of methanol-utilizing bacteria and green algae as predominant members. From these enrichment cultures, two strains of bacteria and two strains of algae were isolated. The microorganisms isolated were designated as bacterium No. 7, bacterium No. 8, Chlorella sp. A-1 and Chlorella sp. B-1, respectively. Stable mixed cultures were easily formed by mixing the isolated cultures of bacteria and algae. Both methanol and bicarbonate were necessary for the growth of the mixed cultures under anaerobic-light conditions. Growth behavior of the mixed cultures was examined on a medium containing 0.1% methanol and 0.1 % bicarbonate at 30°C in the light (about 6000 lx). The maximum specific growth rate for the cultures, µ_max, was 0.092 hr^?1 (doubling time, 7.5 hr). The maximum cell yield was 0.87 g dry-cell weight per g of methanol used. The protein content of the biomass was 65%.