Kinetics of growth and elemental sulfur oxidation in batch culture of thiobacillus ferrooxidans

The kinetics of oxidation of elemental sulfur by Thiobacillus ferrooxidans in a batch reactor was followed by measuring the concentration of adsorbed cells on the sulfur surface, the concentration of free cells in liquid medium, and the amount of sulfur oxidized. As the elemental sulfur was oxidized to sulfate, the liquid-phase concentration of free cells continued to increase with time, whereas the surface concentration of adsorbed cells per unit weight of sulfur approached a limiting value, i.e., the maximum adsorption capacity. During sulfur oxidation, there was a close correlation between the concentrations of adsorbed and free cells, and these data were well correlated with the Langmuir isotherm. The observed rates of batch growth and sulfur oxidation were consistent with a kinetic model, assuming that the growth rate of batch growth and sulfur oxidation were consistent with a kinetic model. Assuming that the growth rate of adsorbed bacteria is proportional to the product of the concentration of adsorbed cells and the fraction of adsorption sites unoccupied by cells. The kinetic and stoichiometric parameters appearing in the model were evaluated using the experimental data and were compared with parameters determined previously for a few metal sulfides. (c) 1994 John Wiley & Sons, Inc.     

Groundwater seepage stimulates the growth of aquatic macrophytes

1. The impact of groundwater seepage on the growth of submerged macrophytes was investigated in experiments on the isoetid Littorella uniflora and the elodeid Myriophyllum alterniflorum both in the laboratory and in the field. Isoetids rely mostly on sediment‐derived CO₂ and nutrients via root uptake, whereas elodeids acquire their inorganic carbon and nutrients from the water column. We thus hypothesised that L. uniflora would respond positively to seeping ground water as it should improve both CO₂ and nutrient supply. 2. Laboratory experiments were conducted by percolating vegetated cores containing natural sediment or technical sand with artificial ground water of high CO₂ concentrations and with either high or low levels of nutrients. Field experiments were conducted in the oligotrophic Lake Hampen, Denmark, with custom‐built seepage‐growth chambers that permitted a near‐natural flow‐through of seeping ground water. Chambers with a solid bottom, and thus no flow‐through of seeping ground water, served as controls in both laboratory and field experiments. In the field, seepage chambers were installed at a site with relatively high seepage fluxes (ground water from forest catchment), at a site with much lower seepage fluxes but with higher nutrient concentrations (ground water from agricultural catchment) and at a reference site with no net discharge or recharge of ground water. 3. Positive growth responses were observed in the field at transects with high groundwater discharge compared to the control chambers with no seepage. No growth response was observed at the reference transect with low or alternating direction of groundwater seepage. The growth rates of L. uniflora in the field were significantly higher in seepage treatments compared to control treatments, and final plant mass was up to 70% higher than that for plants where seepage was excluded. In areas with high groundwater discharge, a strong positive correlation was found between groundwater seepage fluxes, growth rates, and final plant mass for L. uniflora, while there was no such relationship at the reference transect. The growth of M. alterniflorum was also significantly affected by groundwater seepage, but to a lesser degree than L. uniflora. Laboratory experiments generally showed the same trend for both L. uniflora and M. alterniflorum, and the positive influence of seeping ground water was apparently related to increased inorganic carbon supply and, to a lesser degree, improved nutrient availability. 4. Groundwater discharge results in enhanced growth of isoetids and to some extent elodeids inhabiting a groundwater‐fed softwater lake. We propose that the shallow dense vegetation present where most of the discharge takes place acts as a biological filter that retains nutrients that otherwise would end up in the water column and could result in increased algal growth.     

Mechanism of microbial flotation using Thiobacillus ferrooxidans for pyrite suppression

Microbial desulfurization might be developed as a new process for the removal of pyrite sulfur from coal sluries such as coal-water mixture (CWM). An application of iron-oxidizing bacterium Thiobacillus ferrooxidans to flotation would shorten the periods of the microbial removal of pyrite from some weeks by leaching methods to a few minutes. The floatability of pyrite in flotation was mainly reduced by T. ferrooxidans itself rather than by other microbial substances in bacterial culture as additive of flotation liquor. Floatability was suppressed within a few seconds by bacterial contact. The suppression was proportional to increasing the number of cells observed between bacterial adhesion and the suppression of floatability. If 25% of the total pyrite surface area covered with the bacteria, pyrite floatability would be completely depressed. Bacteria that lost their iron-oxidizing activities by sodium cyanide treatment were also able to adhere to pyrite and reduced pyrite floatability as much as normal bacteria did. Thiobacillus ferrooxidans ATCC 23270, T-1, 9, and 11, which had different iron-oxidizing abilities, suppressed floatability to similar-levels. The oxidizing ability of bacteria did not influence the suppressing effect. These results showed the mechanism of the suppression of pyrite floatability by bacteria. Quick bacterial adhesion to pyrite induced floatability suppression by changing the surface property from hydrophobic. The quick adhesion of the bacterium was the novel function which worked to change the surface property of pyrite to remove it from coal. (c) 1993 John Wiley & Sons, Inc.     

Modeling of microbial substrate conversion,growth and product formation in a recycling fermentor

Paracoccus denitrificans and Bacillus licheniformis were grown in a carbon- and energy source-limited recycling fermentor with 100% biomass feedback. Experimental data for biomass accumulation and product formation as well as rates of carbon dioxide evolution and oxygen consumption were used in a parameter optimization procedure. This procedure was applied on a model which describes biomass growth as a linear function of the substrate consumption rate and the rate of product formation as a linear function of the biomass growth rate. The fitting procedure yielded two growth domains for P. denitrificans. In the first domain the values for the maximal growth yield and the maintenance coefficient were identical to those found in a series of chemostat experiments. The second domain could be described best with linear biomass increase, which is equal to a constant growth yield. Experimental data of a protease producing B. licheniformis also yielded two growth domains via the fitting procedure. Again, in the first domain, maximal growth yield and maintenance requirements were not significantly different from those derived from a series of chemostat experiments. Domain 2 behaviour was different from that observed with P. denitrificans. Product formation halts and more glucose becomes available for biomass formation, and consequently the specific growth rate increases in the shift from domain 1 to 2. It is concluded that for many industrial production processes, it is important to select organisms on the basis of a low maintenance coefficient and a high basic production of the desired product. It seems less important that the maximal production becomes optimized, which is the basis of most selection procedures.     

Methane as a carbon substrate for the production of microbial cells

The cultivation of aerobic, methane-utilizing, microbial cells by submerged culture techniques, in an entirely mineral salts medium, with a view to their use as an edible protein source is discussed. Particular emphasis is placed on the potentially explosive nature of gaseous mixtures containing methane and oxygen. The experiments described investigate if fully safe operation at all times, by oxygen concentration control, is possible in agitated and sparged batch fermentors. Appreciable wastage of methane is prevented by gaseous-phase recirculation. It is concluded that fully safe operation is possible, cultures being able to grow exponentially without substrate limitation by the gaseous-phase nutrients.     

Connection between stochastic and deterministic modelling of microbial growth

We present in this paper various links between individual and population cell growth. Deterministic models of the lag and subsequent growth of a bacterial population and their connection with stochastic models for the lag and subsequent generation times of individual cells are analysed. We derived the individual lag time distribution inherent in population growth models, which shows that the Baranyi model allows a wide range of shapes for individual lag time distribution. We demonstrate that individual cell lag time distributions cannot be retrieved from population growth data. We also present the results of our investigation on the effect of the mean and variance of the individual lag time and the initial cell number on the mean and variance of the population lag time. These relationships are analysed theoretically, and their consequence for predictive microbiology research is discussed.     

Determination of the Monod substrate saturation constant for microbial growth

Abstract Methods used to determine the Monod substrate saturation constant for microbial growth are surveyed. The preferred and most accurate method is to assay the concentrations of growth rate-limiting nutrients in steady-state continuous cultures. But, this is not always possible due to the lack of sufficiently sensitive assay methods or due to high nutrient fluxes in rapidly growing cultures. It is suggested that an acceptable and simple alternative method for aerobic microorganisms is to measure initial oxygen uptake rates during growth in the presence of different initial concentrations of growth rate-limiting nutrient. It is important in this method that the microbial cells are taken from rapidly growing cultures and are suspended in a medium permitting growth.     

Marine bacterial diversity as a resource for novel microbial products

Marine bacteria are an important and relatively unexplored resource for novel microbial products. In this review, we discuss a number of issues relevant to the industrial potential of marine microorganisms including how marine and terrestrial bacteria differ, both physiologically and taxonomically, and what constitute reasonable expectations of the biosynthetic capabilities of marine bacteria relative to terrestrial bacteria and to marine macroorganisms. Also discussed is the concept that bacterial associations with marine plants and animals, which range from casual encounters to obligate symbioses, provide unique opportunities for bacterial adaptation. It is proposed that some of these adaptations would not be selected for in the absence of environmental parameters associated with the host, and that these adaptations can include the biosynthesis of unique metabolic products.     

Metasecretome-selective phage display approach for mining the functional potential of a rumen microbial community

Background

In silico, secretome proteins can be predicted from completely sequenced genomes using various available algorithms that identify membrane-targeting sequences. For metasecretome (collection of surface, secreted and transmembrane proteins from environmental microbial communities) this approach is impractical, considering that the metasecretome open reading frames (ORFs) comprise only 10% to 30% of total metagenome, and are poorly represented in the dataset due to overall low coverage of metagenomic gene pool, even in large-scale projects.

Results

By combining secretome-selective phage display and next-generation sequencing, we focused the sequence analysis of complex rumen microbial community on the metasecretome component of the metagenome. This approach achieved high enrichment (29 fold) of secreted fibrolytic enzymes from the plant-adherent microbial community of the bovine rumen. In particular, we identified hundreds of heretofore rare modules belonging to cellulosomes, cell-surface complexes specialised for recognition and degradation of the plant fibre.

Conclusions

As a method, metasecretome phage display combined with next-generation sequencing has a power to sample the diversity of low-abundance surface and secreted proteins that would otherwise require exceptionally large metagenomic sequencing projects. As a resource, metasecretome display library backed by the dataset obtained by next-generation sequencing is ready for i) affinity selection by standard phage display methodology and ii) easy purification of displayed proteins as part of the virion for individual functional analysis.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-356) contains supplementary material, which is available to authorized users.     

The influence of substrate organic content on the growth of a stream chironomid

Stream detritus and Tipula (Diptera) feces were wet-sieved to a 63–250 µm particle size range and mixed in three proportions with ashed sand of the same size to yield food substrates of varying quality. Growth of fourth-instar Stictochironomus annulicrus (Townes) (Diptera: Chironomidae) larvae was used to indicate whether the more nutritious particles were selected from these mixtures. Respective mean relative growth rates on the substrates, in order of decreasing percentage organic matter (i.e. food quality), were: 0.0229, 0.0104 and 0.0004 mg·mg^–1d^–1. Replicates of the relatively high quality substrate consistently elicited the highest growth rates; animals grew at intermediate rates on medium quality, and slowest on low quality substrates. Thus, this collector does not appear to have selected for food quality under the described experimental conditions.     

Quantitative analysis of cellulose degradation and growth of cellulolytic bacteria in the rumen

Ruminant animals digest cellulose via a symbiotic relationship with ruminal microorganisms. Because feedstuffs only remain in the rumen for a short time, the rate of cellulose digestion must be very rapid. This speed is facilitated by rumination, a process that returns food to the mouth to be rechewed. By decreasing particle size, the cellulose surface area can be increased by up to 10⁶-fold. The amount of cellulose digested is then a function of two competing rates, namely the digestion rate ( K _d) and the rate of passage of solids from the rumen ( K _p). Estimation of bacterial growth on cellulose is complicated by several factors: (1) energy must be expended for maintenance and growth of the cells, (2) only adherent cells are capable of degrading cellulose and (3) adherent cells can provide nonadherent cells with cellodextrins. Additionally, when ruminants are fed large amounts of cereal grain along with fiber, ruminal pH can decrease to a point where cellulolytic bacteria no longer grow. A dynamic model based on stella ^® software is presented. This model evaluates all of the major aspects of ruminal cellulose degradation: (1) ingestion, digestion and passage of feed particles, (2) maintenance and growth of cellulolytic bacteria and (3) pH effects.     

Shifts in microbial populations in Rusitec fermenters as affected by the type of diet and impact of the method for estimating microbial growth (15N v. microbial DNA)

Rusitec fermenters are in vitro systems widely used to study ruminal fermentation, but little is known about the microbial populations establishing in them. This study was designed to assess the time evolution of microbial populations in fermenters fed medium- (MC; 50% alfalfa hay : concentrate) and high-concentrate diets (HC; 15 : 85 barley straw : concentrate). Samples from solid (SOL) and liquid (LIQ) content of fermenters were taken immediately before feeding on days 3, 8 and 14 of incubation for quantitative polymerase chain reaction and automated ribosomal intergenic spacer analysis analyses. In SOL, total bacterial DNA concentration and relative abundance of Ruminococcus flavefaciens remained unchanged over the incubation period, but protozoal DNA concentration and abundance of Fibrobacter succinogenes, Ruminococcus albus and fungi decreased and abundance of methanogenic archaea increased. In LIQ, total bacterial DNA concentration increased with time, whereas concentration of protozoal DNA and abundance of methanogens and fungi decreased. Diet×time interactions were observed for bacterial and protozoal DNA and relative abundance of F. succinogenes and R. albus in SOL, as well as for protozoal DNA in LIQ. Bacterial diversity in SOL increased with time, but no changes were observed in LIQ. The incubated diet influenced all microbial populations, with the exception of total bacteria and fungi abundance in LIQ. Bacterial diversity was higher in MC-fed than in HC-fed fermenters in SOL, but no differences were detected in LIQ. Values of pH, daily production of volatile fatty acids and CH₄ and isobutyrate proportions remained stable over the incubation period, but other fermentation parameters varied with time. The relationships among microbial populations and fermentation parameters were in well agreement with those previously reported in in vivo studies. Using ¹⁵N as a microbial marker or quantifying total microbial DNA for estimating microbial protein synthesis offered similar results for diets comparison, but both methods presented contrasting results for microbial growth in SOL and LIQ phases. The study showed that fermentation parameters remained fairly stable over the commonly used sampling period (days 8 to 14), but shifts in microbial populations were detected. Moreover, microbial populations differed markedly from those in the inocula, which indicates the difficulty of directly transposing results on microbial populations developed in Rusitec fermenters to in vivo conditions.     

Integrated biological process for the treatment of a Chilean complex gold ore

Abstract: A process for gold recovery from a complex Chilean ore from Burladora (IV Region) which integrates concentration by flotation, bacterial leaching and cyanidation was studied at a laboratory scale. The chemical composition of the ore is 8.2% Fe, 0.78% Cu, 0.88% As and 3.5 g/t Au, with pyrite, hematite, covelite, arsenopyrite and chalcopyrite as the main metal-bearing minerals. The initial gold recovery by conventional cyanidation on a crushed ore sample was only 54%. The ore was ground and concentrated by flotation with a gold recovery of only 56%. The gold content of the concentrate is 17 g/I Au. Concentrate samples were leached in 1.5 l stirred reactors at 10% pulp density in 1000 ml of acid medium (pH 1.8). Some experiments were inoculated with harvested bacteria previously isolated from mining solutions. Dissolved metals, pH and bacteria concentration in the leaching solutions were periodically determined. In the presence of bacteria, oxidation of the ferrous ion produced by acid dissolution of the concentrate was observed, and after 4 days of leaching 100% of the dissolved iron was present as ferric ion. Gold recovery by cyanidation increased from 13% for the initial concentrate to 34% after 10 days of chemical acid leaching and 97% after 10 days of bacterial leaching. To increase the total gold recovery, the flotation tailings were submitted to cyanidation. A complete flowsheet of the process and a first economical evalualion are proposed. As a possible alternative process, heap bacterial leaching and further cyanidation of the ore are suggested.     

The gram-negative bacterium Escherichia coli as a model for testing the effect of carbonic anhydrase inhibition on bacterial growth

Carbonic anhydrases, catalysing the reversible CO₂ hydration reaction, contribute in all living organisms to the maintenance of stable metabolic functions depending on intracellular concentrations of carbon dioxide, bicarbonate, and protons. Recent studies have examined how CAs affect bacterial lifecycle, considering these enzymes druggable targets due to interference with their activities by using inhibitors or activators. Here, we propose Escherichia coli cells as a model for testing the effect of acetazolamide (AZA), a potent CA inhibitor, on bacterial survival by evaluating E. coli growth through its glucose consumption. AZA, at concentrations higher than 31.2 µg/mL, was able to impair E. coli growth and glucose uptake. AZA is a good inhibitor of the two recombinant E. coli CAs, the β-CA CynT2, and the γ-CA EcoCAγ, with KIs of 227 and 248 nM, respectively. This study provides a proof-of-concept, low-cost method for identifying effective CA inhibitors capable of impairing bacterial metabolism.     

Evaluation of waste chicken feathers as peptone source for bacterial growth

Aims: Peptones are one of the most expensive constituents of microbial media. This study was undertaken to prepare the peptone from waste chicken feathers through a new process. Methods and Results: The chemical analysis of chicken feather peptone (CFP) was performed. The ability of CFP to support the growth of the three test bacteria in liquid and agar media was comparable to those of three commercial peptones [tryptone peptone (TP), fish peptone and protease peptone (PP)]. Conclusions: CFP was found to be rich in ash (42·1 g 100 g^?1), protein (55·8 g 100 g^?1) and mineral contents. The maximum biomass yield (3·13 g l^?1) and colony number (83 × 10⁸ CFU ml^?1) for bacterium Bacillus subtilis were attained with CFP. The maximum biomass yields and colony numbers for Lactobacillus delbrueckii ssp. bulgaricus and Escherichia coli were reached in TP medium. Second high biomass yield (2·64 g l^?1) and colony number (75 × 10⁸ CFU ml^?1) for E. coli were achieved using CFP. Third high biomass yield (1·29 g l^?1) and colony number (90 × 10⁷ CFU ml^?1) for Lact. delbrueckii ssp. bulgaricus were obtained in CFP medium. Significance and Impact of the Study: Usability of waste chicken feathers as substrate for bacteria was investigated for the first time in the present study. The peptone may be used in industrial fermentations for production of antibiotics, organic acids, enzymes and biopolymer. It may be also used in clinical microbiology. A new chemical process was developed for peptone preparation. This process may be also employed for peptone preparation from other organic materials, especially fibrose protein‐containing materials.     

Annual growth layers as proxies of past growth conditions for benthic microbial mats in a perennially ice-covered Antarctic lake

Perennial microbial mats can be the dominant autotrophic community in Antarctic lakes. Their seasonal growth results in clearly discernible annual growth layering. We examined features of live microbial mats from a range of depths in Lake Hoare, Antarctica, that are likely to be preserved in these layers to determine their potential as proxies of past growth performance. Cyanobacteria dominated the mat for all but the deepest depth sampled. Changes in areal concentrations of phycobilin pigments, organic matter and extracellular polysaccharide and in species composition did not correspond to changes in various water column properties, but showed a linear relationship with irradiance. Carbonate accumulation in the mats correlated with biomass markers and may be inferred as an index of mat performance. We examined the carbonate content of annual layers laid down from 1958–1959 to 1994–1995 in sediment cores from 12 m depth. The carbonate content in the layer showed a significant correlation with the mean summer air temperature. These data suggest a link between air temperature and microbial mat growth performance, and suggest that it is mediated via irradiance. Laminated microbial mats in Antarctic lakes have the potential to act as fine-resolution records of environmental conditions in the recent past, although interpretation is complex.     

Dimensionless analysis of the microbial growth rate dependence on sub-optimal temperatures

The influence of sub-optimal temperatures (T) on the microbial growth rate (μ) has been assessed by means of dimensionless variables: T_dim = [T−T_min]/[T_opt−T_min] and μ_dim = μ/μ_opt. T_min represents the temperature at which there is no growth, T_opt the optimum temperature at which the growth rate, μ_opt, is maximum. Data sets, growth rate vs temperature, have been taken from the literature for 12 organisms (psychrotrophs, mesophiles and thermophiles). In order to compare these organisms, the power law function has been used: [μ_dim] = [T_dim]^α. The parameters μ_opt and T_opt are determined from direct readings whereas T_min and αare estimated by means of a non-linear regression. An accurate estimation of T_min is obtained providing low growth rate data are available. A wide range of optimal temperatures where the growth rate almost equals μ_opt prevents one from obtaining a narrow confidence interval forα. On the basis of the analysis hereafter developed, thermophiles are characterized by values of the power α less than mesophiles and psychrotrophs. Almost all of these values are significantly different from two, previously determined for Staphylococcus xylosus and widely used for predicting the microbial growth in foods. Received 15 May 1998/ Accepted in revised form 25 September 1998