Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system

Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy. The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice. Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age. The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass. Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate. Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface. The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate. Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation. (C) 2003 Wiley Periodicals, Inc.

Selection of a strain of Aspergillus for the production of citric acid from pineapple waste in solid-state fermentation

Aspergillus foetidus ACR I 3996 (=FRR 3558) and three strains of Aspergillus niger ACM 4992 (=ATCC 9142), ACM 4993 (=ATCC 10577), ACM 4994 (=ATCC 12846) were compared for the production of citric acid from pineapple peel in solid-state fermentation. A. niger ACM 4992 produced the highest amount of citric acid, with a yield of 19.4 g of citric acid per 100 g of dry fermented pineapple waste under optimum conditions, representing a yield of 0.74 g citric acid/g sugar consumed. Optimal conditions were 65% (w/w) initial moisture content, 3% (v/w) methanol, 30 degrees C, an unadjusted initial pH of 3.4, a particle size of 2 mm and 5 ppm Fe2+. Citric acid production was best in flasks, with lower yields being obtained in tray and rotating drum bioreactors.

O-2 uptake during solid-state fermentation in a rotating drum bioreactor

The maximum O-2 uptake by Rhizopus oligosporus grown in a 200 litre rotating drum bioreactor at 0.5 rpm ranged from 6.7 to 7.6 mmol per min per kg initial dry substrate (IDS), for runs done with 4 baffles each 17 cm wide, and 12 baffles each 5 cm wide. Without baffles, the maximum O-2 uptake rate at 5 rpm was 6.9 mmol/(min.kg IDS), compared to 5.1 mmol/(min.kg IDS) obtained at 0.5 rpm. Therefore O-2 supply is adequate in rotating drum bioreactors as long as slumping flow regimes of the substrate bed are avoided.

Approach to designing rotating drum bioreactors for solid-state fermentation on the basis of dimensionless design factors

The development of large-scale solid-stale fermentation (SSF) processes is hampered by the lack of simple tools for the design of SSF bioreactors. The use of semifundamental mathematical models to design and operate SSF bioreactors can be complex. In this work, dimensionless design factors are used to predict the effects of scale and of operational variables on the performance of rotating drum bioreactors. The dimensionless design factor (DDF) is a ratio of the rate of heat generation to the rate of heat removal at the time of peak heat production. It can be used to predict maximum temperatures reached within the substrate bed for given operational variables. Alternatively, given the maximum temperature that can be tolerated during the fermentation, it can be used to explore the combinations of operating variables that prevent that temperature from being exceeded. Comparison of the predictions of the DDF approach with literature data for operation of rotating drums suggests that the DDF is a useful tool. The DDF approach was used to explore the consequences of three scale-up strategies on the required air flow rates and maximum temperatures achieved in the substrate bed as the bioreactor size was increased on the basis of geometric similarity. The first of these strategies was to maintain the superficial flow rate of the process air through the drum constant. The second was to maintain the ratio of volumes of air per volume of bioreactor constant. The third strategy was to adjust the air flow rate with increase in scale in such a manner as to maintain constant the maximum temperature attained in the substrate bed during the fermentation. (C) 2000 John Wiley & Sons, Inc.

Use of confocal scanning laser microscopy to measure the concentrations of aerial and penetrative hyphae during growth of Rhizopus oligosporus on a solid surface

In order to develop a method for use in investigations of spatial biomass distribution in solid-state fermentation systems, confocal scanning laser microscopy was used to determine the concentrations of aerial and penetrative biomass against height and depth above and below the substrate surface, during growth of Rhizopus oligosporus on potato dextrose agar. Penetrative hyphae had penetrated to a depth of 0.445 cm by 64 h and showed rhizoid morphology, in which the maximum biomass concentration, of 4.45 mg dry wt cm(-3), occurred at a depth of 0.075 cm. For aerial biomass the maximum density of 39.54 mg dry wt(-3) occurred at the substrate surface. For both aerial and penetrative biomass, there were two distinct regions in which the biomass concentration decayed exponentially with distance from the surface. For aerial biomass, the first exponential decay region was up to 0.1 cm height. The second region above the height of 0.1 cm corresponded to that in which sporangiophores dominated. This work lays the foundation for deeper studies into what controls the growth of fungal hyphae above and below the surfaces of solid substrates. (C) Wiley Periodicals, Inc.

Modelling fungal growth on surfaces

Fungal growth in time and space at the substrate surface was modelled for a simple system mimicking solid-state fermentation, using a polycarbonate Nucleopore membrane laid over a glucose solution. Biomass production depends on both tip density and the diffusion of glucose within the fungal hyphae. The model predicts early increases in both height and concentration, followed by a period in which the biomass profile moves with a constant wavefront. The rate of increase in height increases as tip diffusivity increases or as the Monod saturation constant for glucose decreases.

Pectinase production by solid fermentation from Aspergillus niger by a new prescription experiment

The Ordos Plateau in China is covered with up to 300,000 ha of peashrub (Caragana) which is the dominant natural vegetation and ideal for fodder production. To exploit peashrub fodder, it is crucially important to optimize the culture conditions, especially culture substrate to produce pectinase complex. In this study, a new prescription process was developed. The process, based on a uniform experimental design, first optimizes the solid substrate and second, after incubation, applies two different temperature treatments (30 degrees C for the first 30 h and 23 degrees C for the second 42 h) in the fermentation process. A multivariate regression analysis is applied to a number of independent variables (water, wheat bran, rice dextrose, ammonium sulfate, and Tween 80) to develop a predictive model of pectinase activity. A second-degree polynomial model is developed which accounts for an excellent proportion of the explained variation (R-2 = 97.7%). Using unconstrained mathematical programming, an optimized substrate prescription for pectinase production is subsequently developed. The mathematical analysis revealed that the optimal formula for pectinase production from Aspergillus niger by solid fermentation under the conditions of natural aeration, natural substrate pH (about 6.5), and environmental humidity of 60% is rice dextrose 8%, wheat bran 24%, ammonium sulfate ((NH4)(2)SO4) 6%, and water 61%. Tween 80 was found to have a negative effect on the production of pectinase in solid substrate. With this substrate prescription, pectinase produced by solid fermentation of A. niger reached 36.3IU/(gDM). Goats fed on the pectinase complex obtain an incremental increase of 0.47 kg day(-1) during the initial 25 days of feeding, which is a very promising new feeding prospect for the local peashrub. It is concluded that the new formula may be very useful for the sustainable development of and and semiarid pastures such as those of the Ordos Plateau. (c) 2005 Elsevier Inc. All rights reserved.

Residence time distributions of gas flowing through rotating drum bioreactors

Residence time distribution studies of gas through a rotating drum bioreactor for solid-state fermentation were performed using carbon monoxide as a tracer gas. The exit concentration as a function of time differed considerably from profiles expected for plug flow, plug flow with axial dispersion, and continuous stirred tank reactor (CSTR) models. The data were then fitted by least-squares analysis to mathematical models describing a central plug flow region surrounded by either one dead region (a three-parameter model) or two dead regions (a five-parameter model). Model parameters were the dispersion coefficient in the central plug flow region, the volumes of the dead regions, and the exchange rates between the different regions. The superficial velocity of the gas through the reactor has a large effect on parameter values. Increased superficial velocity tends to decrease dead region volumes, interregion transfer rates, and axial dispersion. The significant deviation from CSTR, plug flow, and plug flow with axial dispersion of the residence time distribution of gas within small-scale reactors can lead to underestimation of the calculation of mass and heat transfer coefficients and hence has implications for reactor design and scaleup. (C) 2001 John Wiley & Sons, Inc.

Proving Temporal Properties of Z Specifications Using Abstraction

This paper presents a systematic approach to proving temporal properties of arbitrary Z specifications. The approach involves (i) transforming the Z specification to an abstract temporal structure (or state transition system), (ii) applying a model checker to the temporal structure, (iii) determining whether the temporal structure is too abstract based on the model checking result and (iv) refining the temporal structure where necessary. The approach is based on existing work from the model checking literature, adapting it to Z.

A nanometre-scale non-periodic structural variation in high temperature superconducting ceramics and the implications for properties

Non-periodic structural variation has been found in the high T-c cuprates, YBa2Cu3O7-x and Hg0.67Pb0.33Ba2Ca2Cu3O8+delta, by image analysis of high resolution transmission electron microscope (HRTEM) images. We use two methods for analysis of the HRTEM images. The first method is a means for measuring the bending of lattice fringes at twin planes. The second method is a low-pass filter technique which enhances information contained by diffuse-scattered electrons and reveals what appears to be an interference effect between domains of differing lattice parameter in the top and bottom of the thin foil. We believe that these methods of image analysis could be usefully applied to the many thousands of HRTEM images that have been collected by other workers in the high temperature superconductor field. This work provides direct structural evidence for phase separation in high T-c cuprates, and gives support to recent stripes models that have been proposed to explain various angle resolved photoelectron spectroscopy and nuclear magnetic resonance data. We believe that the structural variation is a response to an opening of an electronic solubility gap where holes are not uniformly distributed in the material but are confined to metallic stripes. Optimum doping may occur as a consequence of the diffuse boundaries between stripes which arise from spinodal decomposition. Theoretical ideas about the high T-c cuprates which treat the cuprates as homogeneous may need to be modified in order to take account of this type of structural variation.

Mass transfer correlations for rotating drum bioreactors

Evaporative cooling is extremely important for large-scale operation of rotating drum bioreactors (RDBs). Outlet water vapour concentrations were measured for a RDB containing wet wheat bran with the aim of determining the mass transfer coefficient for evaporation from the bran bed to the headspace. Mass transfer was expressed as the mass transfer coefficient times the area for transfer per unit volume of void space in the drum. Values of ka' were determined under combinations of aeration superficial velocities ranging from 0.006 to 0.017 ms(-1) and rotation rates ranging from 0 to 9 rpm. Mass transfer coefficients were evaluated using a variety of residence time distributions (RTDs) for flow in the gas phase including plug flow and well-mixed and a Central Jet RTD based on RTD studies. If plug flow is assumed, the degree of holdup at low effective Peclet (Pe(eff)) numbers gives an apparent under-estimate of ka' compared with empirical correlations. Values of ka' calculated using the Central Jet RTD agree well with values of ka' from literature correlations. There was a linear relationship between ka' and effective Peclet number: ka' = 2.32 x 10(-3) Pe(eff). (C) 2002 Elsevier Science B.V. All rights reserved.

The effect of finite bandwidth squeezed light on entanglement creation in the Dicke model

We analyse the relation between local two-atom and total multi-atom entanglements in the Dicke system composed of a large number of atoms. We use concurrence as a measure of entanglement between two atoms in the multi-atom system, and the spin squeezing parameter as a measure of entanglement in the whole n-atom system. In addition, the influence of the squeezing phase and bandwidth on entanglement in the steady-state Dicke system is discussed. It is shown that the introduction of a squeezed field leads to a significant enhancement of entanglement between two atoms, and the entanglement increases with increasing degree of squeezing and bandwidth of the incident squeezed field. In the presence of a coherent field the entanglement exhibits a strong dependence on the relative phase between the squeezed and coherent fields, that can jump quite rapidly from unentangled to strongly entangled values when the phase changes from zero to pi. We find that the jump of the degree of entanglement is due to a flip of the spin squeezing from one quadrature component of the atomic spin to the other component when the phase changes from zero to pi. We also analyse the dependence of the entanglement on the number of atoms and find that, despite the reduction in the degree of entanglement between two atoms, a large entanglement is present in the whole n-atom system and the degree of entanglement increases as the number of atoms increases.

Synthesis and stabilization of nanocrystalline zirconia with MSU mesostructure

In the presence of nonionic block-copolymer surfactant, nanocrystalline zirconia particles with MSU mesostrucmre were synthesized by a novel solid-state reaction route. The zirconia particles possess a nanocrystalline pore wall, which renders higher thermal stability compared to an amorphous framework. To further enhance its stability, laponite, a synthetic clay, was introduced. Laponite acts as an inhibitor to crystal a growth and also as a hard template for the mesostructure. High surface area and ordered pore structure were observed in the stabilized zirconia. The results show that the formation of the MSU structure is attributed to reverse hexagonal micelles, which are the products of the cooperative self-assembly of organic and inorganic species in the solid-state synthesis system with crystalline water and hygroscopic water present.

Mesoporous nanocrystalline zirconia with lamellar and MSU structure

Without introduction of any stabilizer, the mesoporous nanocrystalline zirconia with lamellar and MSU structure was obtained via solid state reaction coupled with surfactant templating method. The phase, surface area and pore structure of zirconia prepared with this novel method could be designed, tailored and controlled by varying synthesis parameters. The phase transformation was controlled by particle size. The mesostructure possesses nanocrystalline pore wall, which renders it more thermally stable than amorphous framework. The results suggest strongly that in solid state synthesis system mesostructure formation still follow the supramolecular self-assembly mechanism. The lamellar and reverse hexagonal structure could be transformed at different OH-/Zr molar ratios in order to sustain the low surface energy of the mesophases. The lamellar structure was preferred at higher OH-/Zr molar ratios but reverse hexagonal was at low ratios.

A model of factors affecting an information system's change in state

Management are keen to maximize the life span of an information system because of the high cost, organizational disruption, and risk of failure associated with the re-development or replacement of an information system. This research investigates the effects that various factors have on an information system's life span by understanding how the factors affect an information system's stability. The research builds on a previously developed two-stage model of information system change whereby an information system is either in a stable state of evolution in which the information system's functionality is evolving, or in a state of revolution, in which the information system is being replaced because it is not providing the functionality expected by its users. A case study surveyed a number of systems within one organization. The aim was to test whether a relationship existed between the base value of the volatility index (a measure of the stability of an information system) and certain system characteristics. Data relating to some 3000 user change requests covering 40 systems over a 10-year period were obtained. The following factors were hypothesized to have significant associations with the base value of the volatility index: language level (generation of language of construction), system size, system age, and the timing of changes applied to a system. Significant associations were found in the hypothesized directions except that the timing of user changes was not associated with any change in the value of the volatility index. Copyright (C) 2002 John Wiley Sons, Ltd.