Kinetic model for selective cultivation of microfungi in a microscreen process for food processing wastewater treatment and biomass production

The research was aimed at developing a technology to combine the production of useful microfungi with the treatment of wastewater from food processing. A recycle bioreactor equipped with a micro-screen was developed as a wastewater treatment system on a laboratory scale to contain a Rhizopus culture and maintain its dominance under non-aseptic conditions. Competitive growth of bacteria was observed, but this was minimised by manipulation of the solids retention time and the hydraulic retention time. Removal of about 90% of the waste organic material (as BOD) from the wastewater was achieved simultaneously. Since essentially all fungi are retained behind the 100 mum aperture screen, the solids retention time could be controlled by the rate of harvesting. The hydraulic retention time was employed to control the bacterial growth as the bacteria were washed through the screen at a short HRT. A steady state model was developed to determine these two parameters. This model predicts the effluent quality. Experimental work is still needed to determine the growth characteristics of the selected fungal species under optimum conditions (pH and temperature).

On systems thinking, systems biology and the in Silico Plant

The recent summary report of a Department of Energy Workshop on Plant Systems Biology (P.V. Minorsky [2003] Plant Physiol 132: 404-409) offered a welcomed advocacy for systems analysis as essential in understanding plant development, growth, and production. The goal of the Workshop was to consider methods for relating the results of molecular research to real-world challenges in plant production for increased food supplies, alternative energy sources, and environmental improvement. The rather surprising feature of this report, however, was that the Workshop largely overlooked the rich history of plant systems analysis extending over nearly 40 years (Sinclair and Seligman, 1996) that has considered exactly those challenges targeted by the Workshop. Past systems research has explored and incorporated biochemical and physiological knowledge into plant simulation models from a number of perspectives. The research has resulted in considerable understanding and insight about how to simulate plant systems and the relative contribution of various factors in influencing plant production. These past activities have contributed directly to research focused on solving the problems of increasing biomass production and crop yields. These modeling approaches are also now providing an avenue to enhance integration of molecular genetic technologies in plant improvement (Hammer et al., 2002).

Summer dormancy in Festuca arundinacea Schreb.; the influence of season of sowing and a simulated mid-summer storm on two contrasting cultivars

Due to the shortage of information on summer dormancy in tall fescue (Festuca arundinacea, syn. Lolium arundinaceum), we tested the response of 2 cultivars of differing dormancy expression and growth stage to a range of summer moisture conditions, including full irrigation, drought, and a simulated mid-summer storm and analysed whether traits associated with summer dormancy conferred better survival under severe field drought. Autumn-sown reproductive and younger, spring-sown plants of 2 cultivars, claimed to exhibit contrasting summer dormancy, were established and then tested in summer 2002 under either long drought, drought+ simulated mid-summer storm, or full irrigation. The autumn-sown reproductive plants of cv. Flecha exhibited traits that can be associated with partial summer dormancy since under summer irrigation they reduced aerial growth significantly and exhibited earlier herbage senescence. Moreover, cv. Flecha used 35% less soil water over the first summer. However, the water status of leaf bases of young vegetative tillers of both cultivars was similar under irrigation and also throughout most of the drought (leaf potential and water content maintained over -4MPa and at approx. 1 g H2O/g DM, respectively). The summer-active cv. Demeter did not stop leaf elongation even in drought and produced twice as much biomass as Flecha under irrigation. Cultivar Demeter responded to the simulated storm with a decline in dehydrin expression in leaf bases, whereas no decline occurred in Flecha, presumably because it remained partially dormant. The younger, spring-sown swards of both cultivars had similar biomass production under summer irrigation but whereas Demeter regrew in response to the simulated storm, cv. Flecha did not, indicating that dormancy, although only partially expressed, was reinforced by summer drought. In all trials, cv. Flecha out-yielded Demeter in autumn regrowth. In particular, the severe drought in 2003 caused a 25% loss of the basal cover in cv. Demeter, whereas Flecha fully maintained its sward allowing it to produce a higher post-drought autumn yield. This work links summer dormancy with higher persistence over long, dry summers.

Nitrogen balance for an agroforestry system irrigated with saline, high nitrogen effluent

Land disposal is commonly used for urban and industrial wastewater, largely due to the high costs involved in alternative treatments or disposal systems. However, the viability of such systems depends on many factors, including the composition of the effluent water, soil type, the plant species grown, growth rate, and planting density. The objective of this study is to establish whether land disposal of nitrogen (N) rich effluent using an agroforestry system is sustainable, and determine the effect of irrigation rate and tree planting density on the N cycle and subsequent N removal. We examined systems for the sustainable disposal of a high strength industrial effluent. The challenge was to leach the salt, by using a sufficiently high rate of irrigation, while simultaneously ensuring that N did not leach from the soil profile. We describe the N balance for two plant systems irrigated with effluent, one comprising Eucalyptus tereticornis and Eucalyptus moluccana and a Rhodes grass (Chloris gayana) pasture, and the other, Rhodes grass pasture alone. Nitrogen balance was assessed from N inputs in effluent and rainfall, accumulation of N in the plant biomass, changes in soil N storage, N loss in run-off water, denitrification and N loss to the groundwater by deep-drainage. Biomass production was estimated from allometric relationships derived from yearly destructive harvesting of selected trees. The N content of that biomass was then calculated from measured N content of the various plant parts, and their mass. Approximately 300 kg N/ha/yr was assimilated into tree biomass at a planting density of 2500 tree/ha of E. moluccana. In addition to tree assimilation, pasture growth between the tree rows, which was regularly harvested, contributed substantially to N uptake. If the trees were harvested after two years of growth and grass harvested regularly, biomass removal of N by the mixed system would be about 700 kg N/ha/yr. The results of this study show that the current system of effluent disposal is not sustainable as the nitrate leaching from the soil profile far exceeds standards set out by the ANZECC guidelines. Hence additional means of N removal will need to be implemented. Biological N removal is an area that warrants further studies as it is aimed at reducing N levels in the effluent before irrigation. This will complement the current agroforestry system.

Nitrogen, phosphorus, silica, and carbon in Moreton Bay, Queensland, Australia: Differential limitation of phytoplankton biomass and production

Subtropical estuaries have received comparatively little attention in the study of nutrient loading and subsequent nutrient processing relative to temperate estuaries. Australian estuaries are particularly susceptible to increased nutrient loading and eutrophication, as 75% of the population resides within 200 km of the coastline. We assessed the factors potentially limiting both biomass and production in one Australian estuary, Moreton Bay, through stoichiometric comparisons of nitrogen (N), phosphorus (P), silicon (Si), and carbon (C) concentrations, particulate compositions, and rates of uptake. Samples were collected over 3 seasons in 1997-1998 at stations located throughout the bay system, including one riverine endmember site. Concentrations of all dissolved nutrients, as well as particulate nutrients and chlorophyll, declined 10-fold to 100-fold from the impacted western embayments to the eastern, more oceanic-influenced regions of the bay during all seasons. For all seasons and all regions, both the dissolved nutrients and particulate biomass yielded N : P ratios < 6 and N : Si ratios < 1. Both relationships suggest strong limitation of biomass by N throughout the bay. Limitation of rates of nutrient uptake and productivity were more complex. Low C : N and C : P uptake ratios at the riverine site suggested light limitation at all seasons, low N : P ratios suggested some degree of N limitation and high N : Si uptake ratios in austral winter suggested Si limitation of uptake during that season only. No evidence of P limitation of biomass or productivity was evident.

Biotechnological production of lactic acid integrated with potato wastewater treatment by Rhizopus arrhizus

This paper describes a feasibility study of a for lactic acid production integrated with are treatment of wastewater from an industrial starch plant. Rhizopus oryzae two strains, Rhizopus arrhizus and Rhizopus oligosporus were tested with respect to their capability to carry out simultaneous saccharification and fermentation to lactic acid using potato wastewater. Rhizopus arrhizus DAR 36017 was identified as a suitable strain that demonstrated a high capacity for starch saccharification and lactic acid synthesis. The optimal conditions, in terms of pH, temperature and starch concentration, for lactic acid production were determined. The selected fungal strain grew well in a pH range from 3.0 to 7.0. The addition of CaCO(3)10 g dm(-3) maintained the pH at 5.0-6.0 and significantly enhanced lactic acid production. Kinetic study revealed that almost complete starch saccharification and a lactic acid yield of 450g kg(-1) could be achieved in 20 h and 28 h cultivation, respectively. The maximum lactic acid production 21 g dm(-3) and mycelial biomass (1.7 g dm(-3)) were obtained at 30degreesC. Besides the multiple bioproducts, total removal of suspended solids and 90% reduction of COD were achieved in a single no-aseptic operation. (C) 2003 Society of Chemical Industry.

Growth, viral production and metabolism of a Helicoverpa zea cell line in serum-free culture

Insect cell cultures have been extensively utilised for means of production for heterologous proteins and biopesticides. Spodoptera frugiperda (Sf9) and Trichoplusia ni (High Five(TM)) cell lines have been widely used for the production of recombinant proteins, thus metabolism of these cell lines have been investigated thoroughly over recent years. The Helicoverpa zea cell line has potential use for the production of a biopesticide, specifically the Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV). The growth, virus production, nutrient consumption and waste production of this cell line was investigated under serum-free culture conditions, using SF900II and a low cost medium prototype (LCM). The cell growth ( growth rates and population doubling time) was comparable in SF900II and LCM, however, lower biomass and cell specific virus yields were obtained in LCM. H. zea cells showed a preference for asparagine over glutamine, similar to the High Five(TM) cells. Ammonia was accumulated to significantly high levels (16 mM) in SF900II, which is an asparagine and glutamine rich medium. However, given the absence of asparagine and glutamine in the medium ( LCM), H. zea cells adapted and grew well in the absence of these substrates and no accumulation of ammonia was observed. The adverse effect of ammonia on H. zea cells is unknown since good production of biologically active HaSNPV was achieved in the presence of high ammonia levels. H. zea cells showed a preference for maltose even given an abundance supply of free glucose. Accumulation of lactate was observed in H. zea cell cultures.

Stable production of hyaluronic acid in Streptococcus zooppidemicus chemostats operated at high dilution rate

Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h(-1)) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose-limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci. (c) 2005 Wiley Periodicals, Inc.

Challenges for simultaneous nitrification, denitrification, and phosphorus removal in microbial aggregates: mass transfer limitation and nitrous oxide production

The microbial community composition and activity was investigated in aggregates from a lab-scale bioreactor, in which nitrification, denitrification and phosphorus removal occurred simultaneously. The biomass was highly enriched for polyphosphate accumulating organisms facilitating complete removal of phosphorus from the bulk liquid; however, some inorganic nitrogen still remained at the end of the reactor cycle. This was ascribed to incomplete coupling of nitrification and denitrification causing NO3- accumulation. After 2 h of aeration, denitrification was dependent on the activity of nitrifying bacteria facilitating the formation of anoxic zones in the aggregates; hence, denitrification could not occur without simultaneous nitrification towards the end of the reactor cycle. Nitrous oxide was identified as a product of denitrification, when based on stored PHA as carbon source. This observation is of critical importance to the outlook of applying PHA-driven denitrification in activated sludge processes. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Correlations between the satellite-derived seasonal cycle of phytoplankton biomass and aerosol optical depth in the Southern Ocean: Evidence for the influence of sea ice

The relationship between the production of dimethylsulfide (DMS) in the upper ocean and atmospheric sulfate aerosols has been confirmed through local shipboard measurements, and global modeling studies alike. In order to examine whether such a connection may be recoverable in the satellite record, we have analyzed the correlation between mean surface chlorophyll (CHL) and aerosol optical depth (AOD) in the Southern Ocean, where the marine atmosphere is relatively remote from anthropogenic and continental influences. We carried out the analysis in 5-degree zonal bands between 50 degrees S and 70 degrees S, for the period ( 1997 - 2004), and in smaller meridional sectors in the Eastern Antarctic, Ross and Weddell seas. Seasonality is moderate to strong in both CHL and AOD signatures throughout the study regions. Coherence in the CHL and AOD time series is strong in the band between 50 degrees S and 60 degrees S, however this synchrony is absent in the sea-ice zone (SIZ) south of 60 degrees S. Marked interannual variability in CHL occurs south of 60 degrees S, presumably related to variability in sea-ice production during the previous winter. We find a clear latitudinal difference in the cross correlation between CHL and AOD, with the AOD peak preceding the CHL bloom by up to 6 weeks in the SIZ. This suggests that substantial trace gas emissions ( aerosol precursors) are being produced over the SIZ in spring ( October - December) as sea ice melts. This hypothesis is supported by field data that record extremely high levels of sulfur species in sea ice, surface seawater, and the overlying atmosphere during ice melt.

Hydrogen production and utilisation opportunities for Australia

Worldwide, research and policy momentum is increasing in the move towards a hydrogen economy. Australia is one of the highest per capita users of energy, but relies heavily on fossil fuels to fulfil its energy requirements-thus making it one of the highest per capita polluters. It is also a country rich in natural resources, giving it the full range of options for a hydrogen economy. With the first Australian Hydrogen Study being completed by the end of 2003, there has as yet been little analysis of the options available to this country specifically. This paper reviews the resources, production and utilisation technology available for a hydrogen economy in Australia, and discusses some of the advantages and disadvantages of the different options. It points out that coal, natural gas, biomass and water are the most promising hydrogen sources at this stage, while solid oxide and molten carbonate fuel cells may hold the advantage in terms of current expertise for utilising hydrogen rich gases for stationary power in Australia. (c) 2004 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.

Adaptive responses of wild mungbean (Vigna radiata ssp sublobata) to photo-thermal environment. II. Growth, biomass, and seed yield

The leaf growth, dry matter production, and seed yield of 11 wild mungbean ( Vigna radiata ssp. sublobata) accessions of diverse geographic origin were observed under natural and artificial photoperiod temperature conditions, to determine the extent to which genotypic differences could be attributed to adaptive responses to photo-thermal environment. Environments included serial sowings in the field in SE Queensland, complemented by artificial photoperiod extension and controlled-environment growth rooms. Photo-thermal environment influenced leaf growth, total dry matter production ( TDM), and seed yield directly, through effects of ( mainly cool) temperature on growth, and indirectly, through effects on phenology. In terms of direct effects, leaf production, leaf expansion, and leaf area were all sensitive to temperature, with implied base temperatures higher than usually observed in cultivated mungbean ( V. radiata ssp. radiata). Genotypic sensitivity to temperature varied systematically with accession provenance and appeared to be of adaptive significance. In terms of the indirect effects of photo-thermal environment, genotypic and environmental effects on TDM were positively related to changes in total growth duration, and harvest index was negatively related to the period from sowing to flowering, similar to cultivated mungbean. However, seed yield was positively related to the duration of reproductive growth, reflecting the indeterminate growth habit of the wild accessions. As a consequence, the wild accessions are more responsive to favourable environments than typically observed in cultivated mungbean, which is determinate in habit. It is suggested that the introduction of the indeterminate trait into mungbean from the wild subspecies would increase the responsiveness of mungbean to favourable environments, analogous to that of black gram ( V. mungo). Although the wild subspecies appeared more sensitive to cool temperature than cultivated mungbean, it may provide a source of tolerance to the warmer temperatures experienced during the wet season in the tropics.

Control of culture environment for improved polyethylenimine-mediated transient production of recombinant monoclonal antibodies by CHO cells

In this study we describe optimization of polyethylenimine (PEI)-mediated transient production of recombinant protein by CHO cells by facile manipulation of a chemically defined culture environment to limit accumulation of nonproductive cell biomass, increase the duration of recombinant protein production from transfected plasmid DNA, and increase cell-specific production. The optimal conditions for transient transfection of suspension-adapted CHO cells using branched, 25 kDa PEI as a gene delivery vehicle were experimentally determined by production of secreted alkaline phosphatase reporter in static cultures and recombinant IgG(4) monoclonal antibody (Mab) production in agitated shake flask cultures to be a DNA concentration of 1.25 mu g 10(6) cells(-1) mL(-1) at a PEI nitrogen: DNA phosphate ratio of 20:1. These conditions represented the optimal compromise between PEI cytotoxicity and product yield with most efficient recombinant DNA utilization. Separately, both addition of recombinant insulin-like growth factor (LR3-IGF) and a reduction in culture temperature to 32 degrees C were found to increase product titer 2- and 3-fold, respectively. However, mild hypothermia and LR3-IGF acted synergistically to increase product titer 11-fold. Although increased product titer in the presence of LR3-IGF alone was solely a consequence of increased culture duration, a reduction in culture temperature post-transfection increased both the integral of viable cell concentration (IVC) and cell-specific Mab production rate. For cultures maintained at 32 degrees C in the presence of LR3-IGF, IVC and qMab were increased 4- and 2.5-fold, respectively. To further increase product yield from transfected DNA, the duration of transgene expression in cell populations maintained at 32 C in the presence of LR3-IGF was doubled by periodic resuspension of transfected cells in fresh media, leading to a 3-fold increase in accumulated Mab titer from similar to 13 to similar to 39 mg L-1. Under these conditions, Mab glycosylation at Asn297 remained essentially constant and similar to that of the same Mab produced by stably transfected GS-CHO cells. From these data we suggest that the efficiency of transient production processes (protein output per rDNA input) can be significantly improved using a combination of mild hypothermia and growth factor(s) to yield an extended activated hypothermic synthesis.