Practical reactor systems for yeast cell immobilization using biomass support particles

The technique of cell immobilization using porous support particles (biomass support particles) has been successfully applied to yeast cells. Two reactor configurations exploiting the use of these particles have been developed and assessed for use in aseptic yeast fermentations. A liquid-fluidized bed fermenter has been devised for use with particles denser than the fermentation liquor whilst a gas-stirred circulating bed fermenter proved suitable for particles of essentially neutral buoyancy. Both systems have been operated successfully for extended periods of continuous operation. The utilization of biomass support particle technology in such reactors provides a practical and robust system for immobilized cell reactors. This technology offers significant opportunities for further development.     

An improved,simple, hydroponic method for growing<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Arabidopsis thaliana is one os the most studied plant model systems. Completing the genomic sequence ofA. thaliana has provided new opportunities for physiological and biochemical studies. While its small size is advantageous for genetic studies, the plant's low biomass makes it difficult to obtain enough plant material for biochemical and physiological research. The small size and rosette leaf structure, combined with the sensitivity of the apical meristem to flooding, make hydroponic growth of this model plant difficult. A few systems for hydroponic culture ofArabidopsis have been described. Gibeaut et al. (1997) introduced the use of rockwool forArabidopsis hydroponic culture. We have improved this system by introducing small-volume plastic containers with improved plugs to support the rockwool. This method is simpler than the original setup and provides improved germination and growth. The smaller containers enable the use of this system in growth chambers or small growth rooms for a large number of parallel experiments.     

Leaf size modifies support biomass distribution among stems, petioles and mid-ribs in temperate plants

The implications of extensive variation in leaf size for biomass distribution between physiological and support tissues and for overall leaf physiological activity are poorly understood. Here, we tested the hypotheses that increases in leaf size result in enhanced whole-plant support investments, especially in compound-leaved species, and that accumulation of support tissues reduces average leaf nitrogen (N) content per unit dry mass (N(M)), a proxy for photosynthetic capacity. Leaf biomass partitioning among the lamina, mid-rib and petiole, and whole-plant investments in leaf support (within-leaf and stem) were studied in 33 simple-leaved and 11 compound-leaved species. Support investments in mid-ribs and petioles increased with leaf size similarly in simple leaves and leaflets of compound leaves, but the overall support mass fraction within leaves was larger in compound-leaved species as a result of prominent rachises. Within-leaf and within-plant support mass investments were negatively correlated. Therefore, the total plant support fraction was independent of leaf size and lamina dissection. Because of the lower N(M) of support biomass, the difference in N(M) between the entire leaf and the photosynthetic lamina increased with leaf size. We conclude that whole-plant support costs are weakly size-dependent, but accumulation of support structures within the leaf decreases whole-leaf average N(M), potentially reducing the integrated photosynthetic activity of larger leaves.     

Abrasion of suspended biofilm pellets in airlift reactors: importance of shape, structure, and particle concentrations

The detachment of biomass from suspended biofilm pellets in three-phase internal loop airlift reactors was investigated under nongrowth conditions and in the presence of bare carrier particles. In different sets of experiments, the concentrations of biofilm pellets and bare carrier particles were varied independently. Gas hold-up, bubble size, and general flow pattern were strongly influenced by changes in volume fractions of biofilm pellets and bare carrier particles. In spite of this, the rate of biomass detachment was found to be linear with both the concentration of biofilm pellets and the bare carrier concentration up to a solids hold-up of 30%. This implies that the detachment rate was dominated by collisions between biofilm pellets and bare carrier particles. These collisions caused an on-going abrasion of the biofilm pellets, leading to a reduction in pellet volume. Breakage of the biofilm pellets was negligible. The biofilm pellets were essentially ellipsoidal, which made three-dimensional size determination necessary. Calculating particle volumes from two-dimensional image analysis measurements and assuming a spherical shape led to serious errors. The abrasion rate was not equal on all sides of the biofilm pellets, resulting in an increasing flattening of the pellets. This flattening was oriented with the basalt carrier inside the biofilm and independent of the absolute abrasion rate. These observations suggest that the collisions causing abrasion are somehow oriented. The internal structure of the biofilms showed two layers, a cell-dense outer layer and an interior with a low biomass density. Taking this density gradient into account, the washout of detached biomass matched observed changes in volume of the biofilm pellets. No gradient in biofilm strength with biofilm depth was indicated. (c) 1997 John Wiley & Sons, Inc.     

On the vegetative biomass partitioning of seed plant leaves, stems, and roots

A central goal of comparative life-history theory is to derive the general rules governing growth, metabolic allocation, and biomass partitioning. Here, we use allometric theory to predict the relationships among annual leaf, stem, and root growth rates (GL, GS, and GR, respectively) across a broad spectrum of seed plant species. Our model predicts isometric scaling relationships among all three organ growth rates: GL is proportional to GS is proportional to GR. It also provides a conceptual basis for understanding the differences in the absolute amounts of biomass allocated to construct the three organ types. Analyses of a large compendium of biomass production rates across diverse seed plant species provide strong statistical support for the predictions of the theory and indicate that reproductive investments may scale isometrically with respect to vegetative organ growth rates. The general rules governing biomass allocation as indexed by the scaling exponents for organ growth rates are remarkably indifferent to plant size and taxonomic affiliation. However, the allometric "constants" for these relationships differ numerically as a function of phenotypic features and local environmental conditions. Nonetheless, at the level of both inter- and intraspecific comparisons, the same proportional biomass allocation pattern holds across extant seed plant species.     

Size–abundance relationships in Florida ant communities reveal how ants break the energetic equivalence rule

1. Ants are among the most abundant terrestrial organisms, yet little is known of how ant communities divide resources because it is difficult to measure the number of individuals in colonies and the density of colonies. 2. The body size–abundance relationships of the ants of five upland ecosystems in Florida were examined. The study tested whether abundance, energy use, and total biomass were distributed among species and body sizes as predicted by Damuth's energetic equivalence rule. Estimates of average worker body size, colony size, colony mass, and field metabolic rates were used to examine the relationships among body sizes, energy use, and total biomass. 3. Analyses revealed significant variation in energy use and did not support the energetic equivalence hypothesis. Specifically, the energy use and total standing biomass of species with large workers and colonies was much greater than smaller species. 4. These results suggest that larger species with larger colonies account for a disproportionate fraction of the total abundance and biomass of ants. A general model of resource allocation in colonies provides a possible explanation for why ants do not conform to the predictions of the energetic equivalence rule and for why ants are so abundant.     

Karyotype variability in successive generations after hybridization between the great sturgeon, Huso huso (L.), and the sterlet, Acipenser ruthenus L.

Effects of particle size, fish size and temperature on the filtration rate of silver carp were determined. When feeding at 20°C on zooplankton and spherical particles (yeast, micronic beads and pollen), 32-g silver carp filter particles larger than 70 urn at a maximum rate of 18.251 h⁻¹. For particles smaller than 70 μm, filtration rates decrease with decreasing particle size until there is no measured filtration for particles smaller than 10 μm. Filtering rates ( FR ) for particles between 10 and 50 μm are described by the equation, FR =−20.8 + 21.7 × log particle diameter. Filtration rates rise as fish size, particle size and temperature increase. Filtration rates per unit biomass, however, fall as fish size increases: FR = 1.54 W^0.713, where FR is the maximum filtration rate in ¹ h ¹ fish ¹ and W is weight of fish in grammes. The results of these trials are consistent with the hypothesis that particle selection by silver carp is a mechanical, passive function of gill raker morphology.     

Hydrodynamic deposition: a novel method of cell immobilization.

A novel method of cell immobilization is described. The cell support consists of ceramic microspheres of approximately 50-75 microns diameter. The spheres are hollow, having a wall thickness of 10-15 microns and one entrance (ca. 20 microns diameter). The walls are porous with a mean pore size of approximately 90 nm. When a cell suspension (of S. cerevisiae) is passed through a column of such particles, cells are immobilized. Conditions are devised such that the overwhelming majority of cells are held in the central cavity of the support and not between the particles. Provided turbulence is avoided, the distribution of cells along the column length in the steady state is rather homogeneous. The facts that (a) essentially all particles, regardless of orientation, entrap cells, and (b) nonporous particles also entrap cells with high efficiency, indicate that filtration effects are irrelevant and that heretofore unrecognized hydrodynamic forces are alone responsible for the cell immobilization. Cells can be immobilized to high biomass densities, while the hydrodynamic properties of columns containing such immobilized cells are excellent. We describe an on-line electronic method for the real-time measurement of immobilized cellular biomass. Cell growth (so recorded) and metabolism continue to occur in such particles at high rates. Using the glycolytic production of ethanol by S. cerevisiae as a model reaction, volumetric productivities as great as any published are obtained. Thus the "lobster-pot effect" or "hydrodynamic deposition" represents a novel, promising, and generally applicable method of cell immobilization.     

Trade-offs in community properties through time in a desert rodent community

Resource limitation represents an important constraint on ecological communities, which restricts the total abundance, biomass, and community energy flux a given community can support. However, the exact relationship among these three measures of biological activity remains unclear. Here we use a simple framework that links abundance and biomass with an energetic constraint. Under constant energetic availability, it is expected that changes in abundance and biomass can result from shifts in the distribution of individual masses. We test these predictions using long-term data from a desert rodent community. Total energy use for the community has not changed directionally for 25 years, but species composition has. As a result, the average body size has decreased by almost 50%, and average abundance has doubled. These results lend support to the idea of resource limitation on desert rodent communities and demonstrate that systems are able to maintain community energy flux in the face of environmental change, through changes in composition and structure.     

南方红壤丘陵区不同土地利用方式下土壤有机碳分布特征及其与草本生物量的关系

不同土地利用方式下不同粒径土壤有机碳含量的变化可以在一定程度上反映土壤碳的变化,对揭示土壤有机碳循环过程具有重要意义.本研究在长期水土流失监测的基础上,采用土壤颗粒分级的方法,以南方红壤丘陵区不同土地利用方式(荒地、松林、草地)坡地土壤为研究对象,探讨了不同土地利用方式对不同粒径土壤有机碳分布特征的影响及其与草本生物量的关系.结果表明:土地利用方式和坡位对不同粒径土壤有机碳含量的影响较明显,研究区不同粒径土壤有机碳含量均表现为草地＞松林＞荒地;不同粒径土壤有机碳所占比例主要取决于土地利用方式,与坡位关系不大;由颗粒有机碳/矿物结合态有机碳(POC/MOC)值可知,草地土壤有机碳较易矿化,而荒地和松林土壤有机碳较稳定;红壤丘陵区坡地土壤砂粒有机碳对草本生物量的影响较大.     

Energy availability, spatial heterogeneity and ecosystem size predict food-web structure in streams

We used standardized techniques to assemble eighteen food webs in streams. Our aim was to identify the determinants of food-web structure with particular reference to energy availability (related to land use), spatial heterogeneity and ecosystem size (both independent of land use). Forested streams displayed lower algal productivity and higher standing crops of organic matter than the grassland streams. The organic matter in the pine streams was probably of lower quality than that elsewhere. Measures of energy availability and spatial heterogeneity predicted species richness and connectance. A combination of energy availability, spatial heterogeneity and ecosystem size accounted for the representation of particular invertebrate feeding groups in the streams. Algal production and organic matter standing crop were important determinants of invertebrate biomass and overall food-web structure. Grassland sites showed a positive relationship between algal productivity and food chain length whereas forest sites displayed a positive relationship between ecosystem size and food chain length. Therefore, these results provide support for both Pimm's productivity hypothesis and Cohen and Newman's ecosystem size hypothesis.     

Biomass segregation in sage cell suspension culture

The biomass of sage (Salvia officinalis L.) cell suspension culture was composed of single cells and cell aggregates. The development of aggregated cell culture from a single-cell suspension was monitored by particle size distribution for four particle size classes. Particle size distribution was compared between the biomass grown in bioreactor and shake flasks. The size of the particles had a strong influence on content of secondary metabolite, ursolic acid (UA). The single cell biomass fraction accumulated up to 7.7 mg UA g^–1 DW which was up to 50 times higher compared to aggregated biomass fractions.     

Biological projectiles (phage, yeast, bacteria) for genetic transformation of plants

Summary Bacteriophage lambda particles, yeast cells, and bacterial cells were tested as projectiles to deliver marker/reporter genes into plant cells via the biolistic process. When phage particles were complexed to tungsten or gold particles and used to bombard tobacco cells, fewer than 15 cell clusters per plate transiently expressed β-glucuronidase (GUS). Cells of wildtype Saccharomyces cerevisiae were too large to be effective projectiles, but use of a reduced-size mutant resulted in a small number of transformants. Escherichia coli cells complexed with tungsten were the most effective projectile for plant transformation. Various methods to prepare E. coli were tested to reduce particle size, improve binding of bacteria to metal particles, and/or minimize particle clumping. In maize, the number of transformants was highest when bacteria/tungsten particles were air-dried onto macrocarriers from an aqueous solution. When maize cells were bombarded with bacteria/tungsten projectiles, rates of transient gene expression (2000 per plate) and stable transformation (50 per plate) were only two- to threefold lower than when purified DNA was used. Transformation of tobacco with E. coli projectiles was improved when the bacteria were treated with a series of ethanol and ether washes, then dried into a powder. Nevertheless, tobacco transformation was still 24- (transient) and 200-fold (stable) less than when purified DNA was used. Biological projectiles can be effective for plant transformation and are advantageous because once a DNA construct is made and put into the appropriate microorganism, the need to isolate and purify DNA for the biolistic process is eliminated, which saves time and lessens DNA shear. Such projectiles may be especially well suited where high molecular weight DNA constructs are needed.     

Co-firing coal with rice husk and bamboo and the impact on particulate matters and associated polycyclic aromatic hydrocarbon emissions

The potential of co-firing rice husk and bamboo with coal was studied in a bench-scale pulverized fuel combustion reactor. Experimental parameters including biomass blending ratio in the fuel mixture, biomass grinding size, excess air ratio and relative moisture content in the biomass were investigated. Particulate Matters in the forms of PM(10), PM(2.1), ultra fine particles as well as the associated Polycyclic Aromatic Hydrocarbons (PAHs) emissions were evaluated. An operation range between 10% and 30% of biomass to coal ratio was found to be the optimum range in terms of minimum pollutant emissions per unit energy output. Co-combustion of coal with biomass seemed to have the effect of moving the fly-ash in PM(2.1) to a larger size range, but increasing the number counts of the ultra fine particles. It was noted that the much higher volatile matter content in the biomass fuels has played a key role in improving the combustion performance in the system. However, slagging, fouling and formation of clinker could be the issues requiring attention when using biomass co-combustion in conventional boilers.     

Seasonal variation in abundance and size structure of phytoplankton in Baie des Chaleurs,southwestern Gulf of St. Lawrence,in relation to physical oceanographic conditions

We investigated changes in the abundance and size structure of phytoplankton and organic seston in relation to temperature, stratification and current patterns at Gascons on the north shore of Baie des Chaleurs, eastern Canada. Phytoplankton biomass showed a general decrease during the study (May to November 1989), except for a brief diatom bloom in late October. During most of the summer, a strong temperature driven stratification was present and <5 µm cells dominated the phytoplankton community. Particles measuring <5 µm also dominated the particulate organic matter (POC and PON) throughout the year. However, only 40% of these particles could be associated with phytoplankton cells. For both particulate matter and phytoplankton, the abundance of the <5 µm size fraction was positively correlated with the Brunt-Väilsälä index of stability of the water column. Inorganic nitrogen may have limited the phytoplankton growth, as generally reported for stratified environments. Most of the biomass was probably supported by nitrogen regenerated through microbial organisms. A large bacterioplankton community was suggested by the abundance of small (<5 µm) non-phytoplanktonic particles with a low and relatively uniform C/N ratio. Larger particles were only abundant at the beginning of the study (May–June) and on one date in October. Their C/N ratios indicated they were of varied origins.     

The production of cellulase in a spouted bed fermentor using cells immobilized in biomass support particles

Continuous cellulase production by Trichoderma viride QM 9123, immobilized in 6 mm diameter, spherical, stainless steel biomass support particles, has been achieved using a medium containing glucose as the main carbon source. Experiments were carried out in a 10-L spouted bed fermentor. In this type of reactor-recycled broth is used to create a jet at the base of a bed of particles, causing the particles to spout and circulate. During the circulation, particles pass through a region of high shear near the jet inlet. This effectively prevents a buildup of excess biomass and thus enables steady-state conditions to be achieved during continuous operation. Continuous production of cellulase was achieved at significantly higher yield and productivity than in conventional systems. At a dilution rate of 0.15 h(-1) (nominal washout rate for freely suspended cells is 0.012 h(-1)), the yield of cellulase on glucose was 31% higher than that measured during batch operation, while the volumetric productivity (31.5 FPA U/L. h) was 53% greater than in the batch system. The specific cellulase productivity of the immobilized cells was more than 3 times that of freely suspended cells, showing that diffusional limitations can be beneficial. This offers significant opportunity for the further development of biomass support particles and associated bioreactors.     

The effects of seed size and pericarp on seedling recruitment and biomass in Cryptocarya alba (Lauraceae) under two contrasting moisture regimes

Establishment success of plants derived from large seeds has been proposed to be greater than that of those derived from smaller ones, particularly under unfavourable conditions of moisture. Therefore, the advantages conferred by large seeds in terms of seedling performance may be modulated by abiotic conditions. The effect of seed size on Cryptocarya alba seedling performance (as determined by seedling recruitment and seedling size) was evaluated under two contrasting rainfall regimes (wet and dry year regime), simulated in the laboratory. It was also determined whether the presence of a pericarp, which had been shown to reduce germination, decreases desiccation and if this counterbalances the greater recruitment of seeds without a pericarp, especially under unfavourable conditions of moisture. Large seeds had a greater probability of recruitment and their seedlings attained a greater biomass, independently of the amount of water applied. In the simulated wet year regime, seeds with a pericarp showed a greater probability of recruitment than those lacking a pericarp. However, seedlings derived from both seed types attained a similar biomass. Under the dry year regime, seeds with and without a pericarp showed similar recruitment probabilities and their seedlings had similar biomasses. These results do not support the assumption that under favourable conditions of moisture, individual differences in seed size would not matter in term of seedling performance. A possible explanation in this case, is the presence of recalcitrant seeds in C. alba, which determines a very short time period for germination following dispersal. Therefore, any attribute that increases germination (e.g., large seeds) would be advantageous, independently of the prevailing abiotic conditions.     

Two-temperature stage biphasic CO2-H2O pretreatment of lignocellulosic biomass at high solid loadings

Most biomass pretreatment processes for monosaccharide production are run at low-solid concentration (<10 wt%) and use significant amounts of chemical catalysts. Biphasic CO(2) -H(2) O mixtures could provide a more sustainable pretreatment medium while using high-solid contents. Using a stirred reactor for high solids (40 wt%, biomass water mixture) biphasic CO(2)-H(2) O pretreatment of lignocellulosic biomass allowed us to explore the effects of particle size and mixing on mixed hardwood and switchgrass pretreatment. Subsequently, a two-temperature stage pretreatment was introduced. After optimization, a short high-temperature stage at 210°C (16 min for hardwood and 1 min for switchgrass) was followed by a long low-temperature stage at 160°C for 60 min. Glucan to glucose conversion yields of 83% for hardwood and 80% for switchgrass were obtained. Total molar sugar yields of 65% and 55% were obtained for wood and switchgrass, respectively, which consisted of a 10% points improvement over those obtained during our previous study despite a 10-fold increase in particle size. These yields are similar to those obtained with other major pretreatment technologies for wood and within 10% of major technologies for switchgrass despite the absence of chemical catalysts, the use of large particles (0.95 cm) and high solid contents (40 wt%).     

Correspondence among methods of zooplankton biomass measurement in lakes: effect of community composition on optical plankton counter and size-fractionated seston data

The effectiveness of the optical particle counter (OPC) to estimatezooplankton biomass depends on the variability in zooplanktonshape and the presence of interfering particles. In marine environmentswhere zooplankton are composed of similarly shaped copepods,an average shape is relatively easily obtained. However, infreshwater environments, spheroid cladocerans mix with ellipsoidcopepods and make the application of a single morphometric modeldifficult. To expand the use of the OPC to freshwater environments,we developed new ellipsoid models for three common lake types(eutrophic, mesotrophic, and oligotrophic). In addition, weassessed how closely different size fractions of seston correspondedto zooplankton biomass. When expressed in common dry mass units,OPC- and seston-derived zooplankton biomass estimates showeda 1:1 correspondence with taxonomically derived estimates inproductive lakes (r > +0.70, P < 0.001) but not in oligotrophicsystems. OPC ellipse models differed among lake sets (major-to-minoraxis ratio: 1.5 to 2.7) but were not a simple function of thecladoceran-to-copepod ratio. The seston size fraction that providedthe best estimates of zooplankton biomass was smaller in mesotrophiclakes (>200 µm) than in eutrophic or oligotrophic lakes(>500 µm). The presence of algae and rotifers had nodetectable influence on OPC and size-fractionated seston estimates.Overall, these analyses suggest that OPC and seston providereliable estimates of lacustrine zooplankton biomass as longas region-specific ellipse models and size fractions, respectively,are used.     

Field study of the relationship between skin-sensitizing antibody production in the cottontail rabbit, Sylvilagus floridanus, and infestation by the rabbit tick, Haemaphysalis leporispalustris (Acri: Ixodidae)

The resistance of cottontail rabbits to tick feeding appears correlated with the rabbits' development of skin-sensitizing antibodies. Resistance appeared to be greatest in adult rabbits which had been repeatedly infested with ticks. Rabbits with little exposure to ticks, usually the young cottontails, showed little or no skin-sensitizing antibody present in their blood and usually had relatively high tick loads when compared with adult rabbits. Models used to interpret the data show promise as tools for predicting tick population fluctuations and, perhaps, incidence of vector borne disease outbreaks. The existence of resistance to tick attachment has important implications for the host-parasite relationship. The research lends support to the hypothesis that the resistance may function as a homeostatic regulatory mechanism capable of maintaining the size of the tick population in equilibrium with the size of the rabbit population. In this way, host resistance may be advantageous to the parasite as well as to the host.