Effect of oxygen on batch and continuous cultures of a nitrogen-fixing Arthrobacter sp.

Growth, acetylene reduction, and respiration rate were studied in batch and continuous cultures of Arthrobacter fluorescents at different oxygen partial pressures. The optimum pO2 values for growth and acetylene reduction were 0.05 and 0.025 atm, respectively, but microorganisms can tolerate higher pO2 values. The growth of cultures provided with combined nitrogen was dependent on oxygen availability, and strict anaerobic conditions did not support growth. Acetylene reduction of a population grown in continuous culture and adapted to low pO2 (0.02 atm) was much more sensitive to oxygenation than that of a population adapted to high pO2 (0.4 atm). Their maximum nitrogenase activity, at their optimal pO2 values, were quite different. The respiratory activity of nitrogen-fixing cultures increased with increasing oxygen tensions until a pO2 of 0.2 atm. At higher pO2 values, the respiration rate began to decrease.     

Influence of chemical pollutants on stability of forest soil microbiocenoses (natural model experiments)

The influence of different concentrations (10, 30, 50, 100, 150, 300) of the maximum permissible concentrations of fluoride and sulfide pollutants (Na₂SO₄, NaF and Na₂SO₃ + NaF) on highly buffered soils of larch forest of Pogorelskii pine wood in Krasnoyarsk forest-steppe was studied. As a result of the influence of treatment with high concentrations of fluoride and sulfide compounds, the intensity of respiration of microorganisms and values of the microbe metabolic coefficient increased, and the biomass of microorganisms and enzymatic activity decreased compared to the control by 1.3–2.7 times. By the end of vegetation, the ecophysiological condition of microbiocenoses of the studied experimental plots had stabilized.     

Optimization of industrial microorganisms: recent advances in synthetic dynamic regulators

Production of biochemicals by industrial fermentation using microorganisms requires maintaining cellular production capacity, because maximal productivity is economically important. High-productivity microbial strains can be developed using static engineering, but these may not maintain maximal productivity throughout the culture period as culture conditions and cell states change dynamically. Additionally, economic reasons limit heterologous protein expression using inducible promoters to prevent metabolic burden for commodity chemical and biofuel production. Recently, synthetic and systems biology has been used to design genetic circuits, precisely controlling gene expression or influencing genetic behavior toward a desired phenotype. Development of dynamic regulators can maintain cellular phenotype in a maximum production state in response to factors including cell concentration, oxygen, temperature, pH, and metabolites. Herein, we introduce dynamic regulators of industrial microorganism optimization and discuss metabolic flux fine control by dynamic regulators in response to metabolites or extracellular stimuli, robust production systems, and auto-induction systems using quorum sensing.     

Changes in phosphatases and lipids following scrotal gamma irradiation in adult rat testis.

Effects of localized low (2.5 Gy) and high (10 Gy) levels of gamma irradiation on the testis of albino rats were studied. A marked increase in the testicular total lipid, phospholipid and cholesterol content was observed at all post-treatment intervals except at 16 weeks where the contents decreased. A significant decrease in the activity of acid phosphatase/g of testis was seen at both the doses, the minimum value being at 4 weeks. The decrease in acid phosphatase activity is correlated with the state of germ cell population in seminiferous tubule which is found to be depleted at 4 week interval. The alkaline phosphatase activity/g testis however, showed a significant increase, the maximum being at 4 weeks post-treatment. Thereafter, the values of the enzyme activity showed a slight recovery at 16 weeks post-irradiation. ATPase activity increased initially followed by a significant decrease at all post-treatment intervals.     

The use of oxygen uptake rate to monitor discovery of microbial and enzymatic biocatalysts

Arising from the requirement for discovery of novel biocatalysts with unusual properties, a process was developed which uniquely combines aspects of continuous culture with the measurement of oxygen uptake. This adaptation of the chemostat can be used to facilitate the isolation of a number of microorganisms with desirable properties, particularly those with useful metabolic capabilities and/or enzymes. The technique was also used to provide feedback on the metabolic status of a microbial population and increase the feed flow rate (i.e., dilution rate) thereby enabling the isolation of microorganisms with enhanced 1,3‐propanediol dehydrogenase activity. The use of oxygen uptake as an indicator of cellular activity enables indirect measurement of substrate utilization and provides a real‐time online assessment of the status of microbial enrichment or evolutionary processes and provides an opportunity, through the use of feedback systems, to control these processes. To demonstrate the utility of the technique, oxygen uptake rate (OUR) was compared with a range of conventional analytical techniques that are typically used to monitor enrichment/evolutionary processes and showed good correlation. Further validation was demonstrated by monitoring a characterizable microbial population shift using OUR. The population change was confirmed using off‐line analytical techniques that are traditionally used to determine microbial activity. OUR was then used to monitor the enrichment of microorganisms capable of using a solvent (1‐methyl‐2‐pyrrolidinone) as the sole source of carbon for energy and biomass formation from a heterogeneous microbial population. After purification the microorganisms taken from the enrichment process were able to completely utilize 1 g L^?1 1‐methyl‐2‐pyrrolidinone within 24 h demonstrating that the technique had correctly indicated the enriched population was capable of growth on 1‐methyl‐2‐pyrrolidinone. The technique improves on conventional microbial enrichment that utilizes continuous culture by providing a real‐time assessment of the enrichment process and the opportunity to use the OUR output for automated control and variation of one or more growth parameters. Biotechnol. Bioeng. 2009;102: 673‐683. © 2008 Wiley Periodicals, Inc.     

Improving a process of cultivation of bifidobacteria in a bioreactor to obtain lyophilized seed culture

The parameters of the cultivation process and the characteristics of the culture growth have been analyzed. The phases of the population growth have been determined. The existence of short stages differing in the physiological state of the cells, their resistance to damaging factors and metabolic activity within the deceleration phase of the growth of bifidobacteria has been experimentally substantiated. To harvest the biomass, the period of the completion of the cultivation process has been chosen. The lyophilized seed culture in which a high percentage of viable cells is preserved for one year has been obtained.     

Microbial seed banks: the ecological and evolutionary implications of dormancy

Dormancy is a bet-hedging strategy used by a wide range of taxa, including microorganisms. It refers to an organism's ability to enter a reversible state of low metabolic activity when faced with unfavourable environmental conditions. Dormant microorganisms generate a seed bank, which comprises individuals that are capable of being resuscitated following environmental change. In this Review, we highlight mechanisms that have evolved in microorganisms to allow them to successfully enter and exit a dormant state, and discuss the implications of microbial seed banks for evolutionary dynamics, population persistence, maintenance of biodiversity, and the stability of ecosystem processes.     

Studies on the toxin of Aspergillus fumigatus. VII. Purification and some properities of hemolytic toxin (asp-hemolysin) from culture filtrates and mycelia.

A hemolytic toxin has been obtained from mycelia and culture filtrates of Aspergillus fumigatus by the procedures that included precipitation with ammonium sulfate, chromatography of DEAE-Sephadex, affinity chromatography on Concanavalin A-Sepharose and gell filtration on Sephadex G-50, G-100 AND G-150. The purified homolytic toxin was homogeneous on immunological and disk electrophoretic analysis, and the toxin from culture filtrates was identical with that from mycelia by the immunodiffusion technique. The hemolytic toxin was obtained for the first time from fungi and designated as Asp-hemolysin. The molecular weight of Asp-hemolysin was estimated to be appoximately 30,000 by the gel-filtration technique and its isoelectric point was found to be around pH 4.0. This Asp-hemolysin contained large amounts of protein and very small amounts of carbohydrate. The UV absorption spectrum of Asp-hemolysin showed a maximum absorption at 280 nm and minimum absorption at 251 nm. The extinction coefficient at 280 nm and minimum absorption at 251 nm. The extinction coefficient at 280 nm, E 1% 1CM, was 12.4 and the ratio of absorbance at 280 nm to that at 260 nm was 2.3. The optimum pH for the hemolytic activity of the toxin toward chicken erythrocytes was 5.0 at room temperature and it was active in the pH range of 3.5 to 10.5. The optimum temperature was 21 C and about 50% of the activity was lost by incubation at 50 C for 5 min or 45 C for 23 min. The hemolytic activity was remarkably inhibited by Hg2+, Cu2+, Fe2+, Ag1+, iodine and p-CMB, but enhanced slightly by Zn2+ and Co2+.     

Thermophilic vs. mesophilic bioleaching process performance

Abstract: Mesophilic and thermophilic mineral oxidising microbial cultures were compared for their capacity to leach both a complex pyritic and an arsenopyritic ore, aiming at copper and gold recovery, respectively. The mesophilic cultures are primarily based on the activity of Thiobacillus -type microorganisms, while the thermophile characteristics place them in the sulphur metabolizers branch df Archaebacteria. Study of key process variables and determination of their optimum values were carried out to provide a basis for significant physiological and technical comparison. Each microbial culture finds a preferential process application, depending on the choice of the reactor and ore.     

BACTERIAL OXIDATION OF DIPICOLINIC ACID I. : Isolation of Microorganisms, Their Culture Conditions, and End Products.

Arima, Kei (University of Tokyo, Tokyo, Japan) and Yasuo Kobayashi. Bacterial oxidation of dipicolinic acid. I. Isolation of microorganisms, their culture conditions, and end products. J. Bacteriol. 84:759-764. 1962.-In a study of the metabolic pathway(s) of dipicolinic acid (DPA) in microorganisms, 436 strains of soil microorganisms were isolated by use of an enrichment culture technique. Most of them were bacteria, and one of them, Achromobacter, which had the strongest DPA-oxidizing activity, was used for the following experiments. In DPA-free medium, the enzymes which oxidize DPA were not produced. The best culture condition for enzyme production and cell growth was: Nutrient Broth supplemented with 0.1% DPA, 30 C, and 20 hr of shake culture. End products were oxalic acid, NH(3), and CO(2). Oxalic acid was not oxidized further by this bacterium. The over-all reaction equation of DPA oxidation was determined.     

Optimal age-dependent sustainable harvesting of natural resource populations: Sustainability value

We studied the optimal age-dependent harvesting of a natural resource population that achieves a maximum income under the constraint of sustainability, i.e. the reproductive adults numbers must exceed a given minimum. The resource species is assumed to be semelparous (a single reproduction over a life). The economic value and natural mortality coefficient can vary with age. The optimal age-dependent harvesting under the sustainability constraint is obtained using Pontryagin’s maximum principle. The constraint of resource sustainability can be treated as an additional term measured in the same units as economic income. Specifically, three terms: (1) current harvesting value, (2) future harvesting value, and (3) sustainability value, are calculated for each age, and the resources should be harvested at the maximum rate when their current harvesting value is greater than the sum of future harvesting value and sustainability value, and should not be harvested otherwise. Numerical analyses of several cases demonstrated that the optimal harvesting schedule depends critically on the natural mortality coefficient and the functional form of the economic value of the resource.     

Diversity of microbial communities in open mixed culture fermentations: impact of the pH and carbon source

Anaerobic fermentation by an open mixed culture was investigated at different pH values (4–8.5) and with three substrates (glucose, glycerol and xylose). The populations established in each condition were assessed by denaturing gradient gel electrophoresis analysis of the 16S ribosomal RNA gene fragments. The fermentation pattern and the composition of the microbial population were also evaluated when operational variations were imposed (increase of substrate concentration or introduction of a second substrate). The experimental results demonstrated that at low and high pH values, a clearly different fermentation pattern was associated with the dominance of a specialised group of clostridiae. At intermediate pH values, the product spectrum was rather variable and seemed to be sensitive to variations in the microbial community. Different substrates resulted in the establishment of different microbial communities. When fed with a mixture of two substrates, mixotrophic microorganisms (capable of degrading both substrates) were found to overgrow the originally dominant specialists. Overall, the experiments have shown that some operational variables have a clear impact on the fermentation pattern and on the population established. However, a uniform relationship between the process characteristics (associated to a metabolic response) and the microbial population present is not always possible. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Device for sterile online measurement of the oxygen transfer rate in shaking flasks

The oxygen transfer rate (OTR) is the most suitable measurable parameter to quantify the physiological state of a culture of aerobic microorganisms since most metabolic activities depend on oxygen consumption. Online measurement of the oxygen transfer rate in stirred bioreactors is state of the art although technically difficult. However, the online determination of the oxygen transfer rate in shaking bioreactors under sterile conditions has not been possible until recently. A newly developed measuring device eliminates this deficit. Extremely useful information about cultivating conditions and the physiological state of microorganisms can be gained in early stages of research and bioprocess development from many reactors operated in parallel.     

A microbiological study of Hussuwa: a traditional Sudanese fermented food from germinated Sorghum bicolor c.v. feterita

Hussuwa is a traditional Sudanese fermented food. Hussuwa made from Sorghum bicolor variety feterita exists in northern, central and eastern Sudan. The microbiological study indicated that the fermentation was primarily a lactic acid fermentation. The changes in microbial population, acetic acid bacteria, lactic acid bacteria and yeasts during all stages of hussuwa preparation and ripening were studied. The identification of fermented hussuwa microorganisms revealed that the main microorganisms were Lactobacillus saccharolyticum, Gluconobacter oxydans, Acetobacter xylinum and Saccharomyces cerevisiae. The metabolic products were studied in all stages of preparation and the period of ripening of hussuwa. The values of pH decreased as fermentation proceeded, and titratable acidity and volatile fatty acids increased.     

The role of glucocorticoids in the regulation of the diurnal rhythm of hepatic beta-hydroxy-beta-methylglutaryl-coenzyme A reductase and cholesterol 7 alpha-hydroxylase.

The microsomal activities of the hepatic enzymes hydroxymethylglutaryl-CoA reductase and cholesterol 7 alpha-hydroxylase exhibit a diurnal rhythm with maximum activities observed during the dark period and minimum activities around noon (12:00h). This diurnal rhythm was maintained for both enzymes after adrenalectomy, but the amplitude of variation for the activity of both enzymes was greatly decreased. A single injection of cortisol administered to adrenalectomized rats 3h before the expected maximum in enzyme activity resulted in a twofold increase in the activity of both enzymes 3h later, at values similar to those observed for control rats killed at the same time. This response appeared to require protein synthesis, since it was blocked by actinomycin D. However, the administration of cortisol to adrenalectomized rats 3 h before the expected minimum did not result in significant change in the activity of hydroxymethylglutaryl-CoA reductase and cholesterol 7 alpha-hydroxylase 3 h later. Kinetic studies of cholic acid metabolism in vivo demonstrated that adrenalectomy results in a significant decrease in the rate of synthesis of cholic acid and a considerable decrease in the pool size of cholic acid and its metabolic products. Treatment of adrenalectomized rats with cortisol increased the rate oonsistent with the effects of adrenalectomy and cortisol treatment on the activity of cholesterol 7alpha-hydroxylase.     

Effect of high-pressure gas on yeast growth

Microcalorimetry is a useful tool for monitoring the growth behavior of microorganisms. In this study, microcalorimetry was used to investigate the effects of nitrogen, air, oxygen, nitrous oxide, argon, and krypton at high pressure on the growth of the yeast Saccharomyces cerevisiae. Growth thermograms (metabolic heat vs. incubation time) were generated to estimate metabolic activity under compressed gases and to determine the 50% inhibitory pressure (IP(50)) and minimum inhibitory pressure (MIP), which are regarded as indices of the toxicity of compressed gases. Based on MIP values, the most toxic to the least toxic gases were found to be: O(2) > N(2)O > air > Kr > N(2) > Ar.     

Einfluß der Kultivierungsbedingungen von Mikroorganismen auf ihre Metallsorption

The silver sorption capacity of three methylotrophic bacteria and one gelatinotrophic Pseudomonas strain has been investigated in dependence of the cultivation conditions. Whereas the maximum loading capacity of the microorganisms was not appreciable influenced by the variation of the dilution rate in the range between 0.05 h^?1 to 0.5 h^?1, a change of the carbon source or the modification of the limitation conditions during the chemostatic cultivation process provokes a strong alteration in the sorption behaviour. The maximum silver loading values were obtained, when the microorganisms are cultivated under N-limitation and the minimum values were measured under P-limitation.     

Effects of culture conditions on production of antibiotically active metabolites by the marine alga Spyridia filamentosa (Ceramiaceae, Rhodophyta). I. Light

The effect of irradiance over the range 5 to 70 μmol photon m-2 s-1 on production of antibiotically active metabolites was assessed for male and female gametophytes and tetrasporophytes of the red alga Sphyridia filamentosa (Wulfen) Harvey in Hooker in culture. Whole-algal extracts and ten recognizable TLC-separable zones were assayed against five human microorganisms pathogenic to humans. For all experimental irradiance conditions, the ten TLC zones displayed activity against four of the microorganisms. The maximum number of TLC zones with activity under any of the culture conditions was six each for male and female Spyridia at 70 μmol photon m-2 s-1. Small changes in irradiance resulted both in different activities against specific microorganisms and degree of activity. The fact that every TLC zone showed differing activities at different light conditions or when extracted from different life history stages strongly suggests the presence of multiple antibiotic principals in individual TLC zones. This revised version was published online in June 2006 with corrections to the Cover Date.     

Is bioaugmentation a feasible strategy for pollutant removal and site remediation?

Microorganisms can degrade numerous organic pollutants owing to their metabolic machinery and to their capacity to adapt to inhospitable environments. Thus, microorganisms are major players in site remediation. However, their efficiency depends on many factors, including the chemical nature and the concentration of pollutants, their availability to microorganisms, and the physicochemical characteristics of the environment. The capacity of a microbial population to degrade pollutants within an environmental matrix (e.g. soil, sediment, sludge or wastewater) can be enhanced either by stimulation of the indigenous microorganisms by addition of nutrients or electron acceptors (biostimulation) or by the introduction of specific microorganisms to the local population (bioaugmentation). Although it has been practiced in agriculture and in wastewater treatment for years, bioaugmentation is still experimental. Many factors (e.g. predation, competition or sorption) conspire against it. However, several strategies are currently being explored to make bioaugmentation a successful technology in sites that lack significant populations of biodegrading microorganisms. Under optimal local conditions, the rate of pollutant degradation might increase upon addition of an inoculant to remediate a chemical spill; however, the most successful cases of bioaugmentation occur in confined systems, such as bioreactors in which the conditions can be controlled to favour survival and prolonged activity of the exogenous microbial population.     

Estimation of vaginal probiotic lactobacilli growth parameters with the application of the Gompertz model

Lactobacilli are widely described as probiotic microorganisms used to restore the ecological balance of different animal or human tracts. For their use as probiotics, bacteria must show certain characteristics or properties related to the ability of adherence to mucosae or epithelia or show inhibition against pathogenic microorganisms. It is of primary interest to obtain the highest biomass and viability of the selected microorganisms. In this report, the growth of seven vaginal lactobacilli strains in four different growth media and at several inoculum percentages was compared, and the values of growth parameters (lag phase time, maximum growth rate, maximum optical density) were obtained by applying the Gompertz model to the experimental data. The application and estimation of this model is discussed, and the evaluation of the growth parameters is analyzed to compare the growth conditions of lactobacilli. Thus, these results in lab experiments provide a basis for testing different culture conditions to determine the best conditions in which to grow the probiotic lactobacilli for technological applications.