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An assessment of seed quality and its influence on productivity estimation in an industrial antibiotic fermentation.

This study investigates the benefits of including seed quality information into data-based models for final productivity estimation in an industrial antibiotic fermentation process. Multiway principal component analysis is applied to assess the seed quality using routinely gathered plant data. Multiway partial least-squares regression is then used to estimate the final productivity using data from the main fermentation only. The issue of selecting appropriate process variables as inputs is investigated. Subsequently, seed characteristics are included into the estimation models to assess the benefits of including information from this stage for productivity estimation. It is shown that it is possible to extract seed fermentation features related to the final productivity both at pilot and production scales. It is postulated that significant influential variations are mirrored in monitored variables during the main fermentation, and therefore seed quality is implicitly accounted for.

Anti-Bacterial Agents↗

An effective method for extracting DNA from contaminated soil due to industrial waste.

Recently, labor accidents caused by toxic gas production such as H2S and CH4 have happened at some waste disposal sites and illegal dumping sites in Japan. The gas production has been known to be end products of bacterial respiration. In order to investigate a microbial flora in industrial waste contaminated soil by a DNA-based detection method, we developed a simple DNA extraction method. The method is based on lysis with sodium dodecyl sulfate (SDS) and a high concentration (4.0 mg/ml) of lysozyme at room temperature. DNA yields increased according to the concentration of lysozyme (0-2 mg/ml), although the cell lysis occurred at comparable efficiency independent of the lysozyme concentration. Interestingly, bovine serum albumin and insulin (4 mg/ml) also contributed high DNA yields. This suggests proteins play an important role at high concentration on a DNA extraction process, presumably by suppressing DNA degradation or adsorption. DNA yields from three contaminated soil samples at 0.3, 1.5 and 3 m in depth by using our method were significantly higher than those extracted by using a widely-used conventional method which did not contain lysozyme. A novel and simple DNA extraction method using high concentration of protein was established for a DNA-based detection method of microbial flora in soil.

Air Pollution↗

Synthesis and analysis of mixed chlorinated-fluorinated dibenzo-p-dioxins and dibenzofurans and assessment of formation and occurrence of the fluorinated and chlorinated-fluorinated dibenzo-p-dioxins and dibenzofurans.

Various mixtures of chlorinated-fluorinated dibenzodioxins (PCFDD), dibenzofurans (PCFDF) and biphenyls (PCFB) were synthesized. For trace analysis, we compared the behavior of PCFDD/PCFDF and chlorinated congeners in extraction, enrichment and clean-up steps. To establish GC/MS analysis, various columns were tested, temperature programs were optimized and for MS-identification specific masses were selected from the mass spectra. To evaluate the possibility of formation of these compounds in thermal processes, samples from the aluminum-producing industry and products of pyrolysis of chlorofluoro- hydrocarbons (FCKW) on a laboratory scale were analyzed. At present, the most important potential sources of fluorinated or mixed chlorinated-fluorinated dioxins, furans and biphenyls - combustion processes and large scale chemical production - do not cause significant environmental contamination at least in the areas considered. However, in a sample of filter dust from the refining process of aluminum using freon 12 we found high amounts (4.5 g/kg !) of chlorinated and chlorinated-fluorinated aromatic compounds including chlorinated-fluorinated dibenzofurans and biphenyls. Since the FCKW ban in Europe, this procedure was modified in 1992, replacing freon 12 by a mixture of chlorine and argon. The PFDD/PFDF concentration in the fluorophenols was between 0.45 and 120 micrograms/kg. The fluorobenzenes analyzed contained between 15 and 70 mg/kg fluorinated biphenyls.

Benzofurans↗

Seasonal variability of total and easily leachable element contents in topsoils (0-5 cm) from eight catchments in the European Arctic (Finland, Norway and Russia).

Frozen topsoil samples (0-5 cm) were collected during March/April 1994 in eight Arctic catchments in northern Europe (4 in Russia, 3 in Finland, 1 in Norway) at varying distances and wind directions from the emissions of the Russian nickel ore mining, roasting and smelting industry on the Kola Peninsula. Between 14 and 25 sites were sampled in catchment basins ranging in size from 12 to 35 km(2). Sampling was repeated in spring immediately after the snow melted, in summer and in autumn to study seasonal variability and the fate of elements when the snow melts. The <2 mm fraction of air-dried topsoils was analysed for total (aqua regia extraction) and easily leachable (in 1 m ammonium acetate, buffered at pH 4.5) element concentrations using ICP-AES and GFAAS for up to 35 elements. Results for selected elements are presented here. Soil organic matter can be shown to be the controlling factor determining element contents and fate. In catchments close to the Russian nickel industry, the topsoils have low carbon and nitrogen contents. Using both extraction methods most elements reach maximum concentrations in winter; lowest concentrations are observed in midsummer. Soil organic matter and elements associated with it are thus leached out of the soils together with soluble elements when the snow melts. This process continues in summer. Elements will enrich surface waters, the lower layers of podzol profiles, or reach the groundwater. The use of the two extractions described provides a simple method to study the mobilities and pathways of elements in the topsoils during the arctic year. Using the proportions of easily leachable to total concentration, a good estimation of the status of the topsoil in the study area can be given.

Journal Article↗

The antioxidative power AP--A new quantitative time dependent (2D) parameter for the determination of the antioxidant capacity and reactivity of different plants.

In the last decade, naturally occurring antioxidants continue to play an important role in the food-supplement industry. The content of antioxidants in a plant depends on the species, temperature, humidity, period of growth, harvest month, part of the plant used and many other variables. Herein, we present a new method able to determine the all over antioxidative power (AP) of plant extracts or lyophilised plant parts based on the reducing activity against a stable test radical. The method is performed by ESR spectroscopy and is based on the well-known 1,1-diphenyl-2-picryl-hydrazil (DPPH) method with the major difference that both the antioxidative capacity and the antioxidative activity are used to characterise an antioxidant. The resulting antioxidative power is expressed in antioxidative units (AU), where 1AU corresponds to the activity of a 1 ppm solution of Vitamin C as a benchmark. This method allows a rapid, unexpensive and general applicable technique for the measurement of the antioxidative power of very different kinds of substances. The inclusion of the kinetic behaviour of the reducing process of the antioxidant for the determination of the AP allows the identification of the main antioxidant present in a sample. Herein, we present the application example of seeds, sprouts and adult parts of dandelion, amaranth, quinoa, fenugreek, broccoli, red clover and mugwort, where the AP method permits to characterise the plants with the highest antioxidant capacity and reaction velocity. The method permits to select active plant extracts for the food and nutrition industry.

Antioxidants↗

Species-specific PCR for the identification of ovine, porcine and chicken species in meta and bone meal (MBM).

BSE, first identified in the UK in 1986 is thought to have arisen from feeding scrapie infected Meat and Bone Meal (MBM), produced under sub-optimal conditions, to cattle. For quality and safety reasons there is a requirement for a good analytical test for the surveillance of processed MBM. This study describes species-specific PCR assays for the identification of ovine, porcine and poultry species in MBM. A comparison between two distinct DNA extraction methods, i.e. the silicaguanidiumthiocyanate DNA isolation procedure and a commercial DNA extraction kit, is also presented. Application of this technology to species identification in industrial MBM was investigates as part of this study.

Animal Feed↗

Effect of environmental factors on the production of oxygenated unsaturated fatty acids from linoleic acids by Bacillus megaterium ALA2.

We identified many novel oxygenated fatty acids produced from linoleic acid by Bacillus megaterium ALA2: 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17-THOA); 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA); 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9(Z)-octadecenoic acid. 12,13,17-THOA, the main product, has antiplant pathogenic fungal activity. To develop an industrial process for the production of these new oxygenated fatty acids by strain ALA2, the effect of environmental factors on the production and their impact on the amount of various products were studied. Dextrose at 5 g/l was the optimum amount for the carbon source. A combination of 15 g yeast extract and 10 g tryptone showed good results as nitrogen sources. Among the metal ions tested, the optimum concentrations for the reaction for the different ions were as follows (in mM): magnesium 2.0, iron 0.5, zinc 0.1, nickel 0.01, and cobalt 0.05. Copper ions did not affect the production of oxygenated products; however, manganese ions inhibited the reaction. Addition of these metal ions did not alter the distribution of products. The optimum temperature and pH for the production of THOAs were 30 degrees C and pH 6.5. Time course studies showed 40-48 h is the optimum for the production of both THOAs. These data provide the basis for engineering scale-up production of these new products.

Aerobiosis↗

Release of mutagens from finished leather.

Extracts of a leather widely used in the furniture and dress-making industries were tested for their mutagenic activity in the Salmonella/microsome assay. Extracts obtained after vigorous treatment of leather samples in a Soxhlet apparatus with toluene or ethanol were mutagenic in strain TA98 of S. typhimurium in the absence of S9 mix. The analysis of extracts of leather at various intermediate stages of processing showed that the mutagenic activity appeared after the coloration process. The responsible compound was identified to be an azo dye (Color Index: Acid Brown 83) whose mutagenic potency was about 4 revertants/micrograms.

Azo Compounds↗

Electrokinetic enhancement removal of heavy metals from industrial wastewater sludge.

An enhanced electrokinetic process for removal of metals (Cr, Cu, Fe, Ni, Pb, Zn) from an industrial wastewater sludge was performed. The electrokinetic experiments were conducted under a constant potential gradient (1.25 V cm(-1)) with processing fluids of tap water (TW), sodium dodecylsulfate (SDS) and citric acid (CA) for 5 days. Results showed that metal removal efficiency of heavy metals for EK-TW, EK-SDS and EK-CA systems are 11.2-60.0%, 37.2-76.5%, and 43.4-78.0%, respectively. A highest metal removal performance was found in EK-CA system. The removal priority of investigated metals from sludge by EK process was found as: Cu > Pb > Ni > Fe > Zn > Cr. The results of sequential extraction analysis revealed that the binding forms of heavy metals with sludge after electrokinetic process were highly depend upon the processing fluid operated. It was found that the binding forms of metals with sludge were changed from the more difficult extraction type (residual and sulfate fractions) to easier extraction types (exchangeable, sorbed, and organic fraction) after treatment by electrokinetic process. Results imply that if a proper treatment technology is followed by this EK process to remove metals more effectively, this treated sludge will be more beneficial for sludge utilization afterwards. Before it was reused, the risk associated with metals of more mobile forms to the environment need to be further investigated. The cost analysis was also evaluated for the investigated electrokinetic systems.

Electrochemistry↗

Production of antibodies and antibody fragments in plants.

Our current knowledge allows the generation of transgenic plants that efficiently produce heterologous proteins from plant, bacterial, fungal or animal origin. Among all types of recombinant proteins, antibodies are particularly attractive because of their ability to specifically recognize and bind virtually any type of antigen. Plants show several advantages as a large-scale antibody production system: they can be grown easily and inexpensively in large quantities that can be harvested, stored and processed by using existing infrastructures. Isolation and purification of plant-made antibodies, if necessary, allow fundamental, industrial, and therapeutical applications. In the past, we and others have successfully generated antibody-producing plants. The maximal accumulation levels of antibodies and antibody fragments that we observed are 1-5% of the extracted proteins. Currently, several biotechnological companies grow field crops to produce antibodies for ex planta applications on an industrial scale.

Animals↗

A simple, two-component buffer enhances use of chromatofocusing for processing of therapeutic proteins.

To extend the feasibility of chromatofocusing to industrial use, we have developed a simple chromatofocusing buffer system capable of generating a smooth pH gradient without the use of an external gradient maker. Using two cationic buffering components, an internal pH gradient is produced on appropriate chromatography media over a broad pH range (9.5 to 5.0). The utility of this buffer system is demonstrated with PBE94 and DEAE Sepharose fast flow ion-exchangers, as well as with experimental fast flow chromatofocusing gels. Using a rapid flow rate, we evaluated this buffer system for recovery of a therapeutic protein from a bacterial cell extract. The simplicity of the buffer system requiring no external gradient maker, coupled with the use of fast flow chromatographic media to produce broad-range pH gradients, improves the scalability of chromatofocusing for processing of therapeutic proteins.

Biotechnology↗

Acoustic demixing of aqueous two-phase systems.

Aqueous two-phase systems demix slowly due to similar physical properties. This is one of the major drawbacks for their adaptation for industrial scale extraction of enzymes. In the present work, a method to accelerate the demixing rates of these systems, employing a traveling acoustic wave field is reported for the first time. Phase-demixing for three systems, viz. polyethylene glycol (PEG)/sodium sulfate, PEG/potassium phosphate and PEG/maltodextrin were studied. The acoustically assisted process decreased the demixing time significantly (about 2- to 3-fold in PEG/salt systems and about 2-fold in the PEG/maltodextrin system), compared to that in gravity alone. Ultrasonication apparently enhanced the coalescence of the dispersed phase droplets due to the mild circulation currents it caused in the dispersion. This in turn enhanced the rate of demixing due to the increased migration velocity of the larger droplets.

Acoustics↗

Acrylamide in French fries: influence of free amino acids and sugars.

The free amino acid profile and sugar (fructose, glucose, and sucrose) composition were determined in potato samples selected to give a large range of variation (a total of 66 samples). From these samples French fries were produced in a laboratory-scale simulation of an industrial process followed by a finish fry at 180 degrees C for 3.5 min using a restaurant fryer. The final product was blast frozen and analyzed for acrylamide. Acrylamide was detected in all samples, but its concentration varied significantly from 50 to 1800 ng/g. For isotope dilution (13C3) acrylamide analysis, samples were extracted with water, cleaned up on HLB Oasis polymeric and Accucat mixed mode anion and cation exchange SPE columns, and analyzed by LC-MS/MS. Statistical analysis of the data indicates that the effect of sugars and asparagine on the concentration of acrylamide in French fries is positive and significant (p < 0.001). It appears that one of the ways acrylamide formation in French fries can be effectively controlled is by the use of raw products with low sugar (and to a lesser degree, asparagine) content.

Acrylamide↗

Remediation of contaminated soils using supercritical fluid extraction: a review (1994-2004).

Considerable effort is being made to remediate soils contaminated with petroleum hydrocarbons, polyaromatic hydrocarbons, polychlorinated biphenyls, dioxins, heavy metals and other organic and inorganic compounds that have resulted from industrial activities, accidental spills and improper waste disposal practices. Current remediation technologies may be limited when treating certain types of contaminated soils and therefore new, efficient and cost effective technologies are being investigated. Supercritical fluid extraction is a potential remediation technology for contaminated soils. It is a simple, fast and selective solvent extraction process that uses a supercritical fluid as the solvent. A commonly used fluid is carbon dioxide at pressures and temperatures greater than 7.4 MPa and 31 degrees C, respectively. In supercritical fluid extraction, the extracted contaminants first dissolve into the supercritical solvent and then these contaminants are separated from the supercritical solvent via a simple change in pressure and temperature conditions or by using a separation process. This paper provides a review of supercritical fluid extraction and its application to the remediation of contaminated soils. This review focuses on the removal of organic contaminants (such as petroleum hydrocarbons, polyaromatic hydrocarbons, polychlorinated biphenyls and others) and inorganic contaminants (such as heavy metals and radioactive elements) from soils. Recent data (1994-2004) on the supercritical fluid extraction of spiked soils and field-contaminated soils were collected. The success of supercritical fluid extraction as a method for removing these contaminants from soils is highlighted and some of the future research needed to develop it as a commercial-scale economic remediation technology are discussed.

Chromatography, Supercritical Fluid↗

Water circuit closure with membrane technology in the pulp and paper industry.

In this study, membrane filtration as an internal purification method, "the kidney", in the pulp and paper industry is discussed. Membrane filtration is economically competitive and a very versatile process. It can be used to remove the enriched organic and/or inorganic loads either partially or totally from, for example, the mechanical pulping and paper making water circuits and it can be applied to various points within the process. With the so-called shear enhanced membrane modules very high fluxes, in ultrafiltration about 400 L/(m2h) and in nanofiltration fluxes almost 200 L/(m2h), have been obtained. Depending on the membrane, suspended solids (microfiltration), polysaccharides, extractives and high molar mass lignous substances (ultrafiltration) and multivalent salts (nanofiltration) can be removed. Ultrafiltration permeate can well be used in paper machine showers to replace fresh water. The quality of the nanofiltration permeate is significantly higher than that of ultrafiltration. The membrane processes can be enhanced by various pre-treatment techniques to produce higher permeate fluxes and to eliminate membrane fouling. Such pre-treatment methods are, e.g., chemical treatment, ozonation and biological treatment. The most cost-effective processes were chemical flocculation, pH adjustment and thermophilic aerobic biological treatment.

Filtration↗

Isomaltulose production using free cells: optimisation of a culture medium containing agricultural wastes and conversion in repeated-batch processes.

The enzyme glucosyltransferase is an industrially important enzyme since it produces non-cariogenic isomaltulose (6-O-alpha-D-glucopyronosyl-1-6-D-fructofuranose) from sucrose by intramolecular transglucosylation. The experimental designs and response surface methodology (RSM) were applied for the optimisation of the nutrient concentrations in the culture medium for the production of glucosyltransferase by Erwinia sp. D12 in shaken flasks at 200 rpm and 30 degrees C. A statistical analysis of the results showed that, in the range studied, the factors had a significant effect (P < 0.05) on glucosyltransferase production and the highest enzyme activity (10.84 U/ml) was observed in culture medium containing sugar cane molasses (150 g l(-1)), corn steep liquor (20 g l(-1)), yeast extract Prodex Lac SD (15 g l(-1)) and K2HPO4 (0.5 g l(-1)) after 8 h at 30 degrees C. The production of cell biomass by the strain of Erwinia sp. D12 was carried out in a 6.6-l fermenter with a mixing rate of 200 rpm and an aeration rate of 1 vvm. Fermentation time, cellular growth, medium pH and glucosyltransferase production were observed. The greatest glucosyltransferase activity was 22.49 U/ml, obtained after 8 h of fermentation. The isomaltulose production from sucrose was performed using free Erwinia sp. D12 cells in a batch process using an orbital shaker. The influence of the parameters sucrose concentration, temperature, pH, and cell concentration on the conversion of sucrose into isomaltulose was studied. The free cells showed a high conversion rate of sucrose into isomaltulose using batch fermentation, obtaining an isomaltulose yield of 72.11% from sucrose solution 35% at 35 degrees C.

Biomass↗

Manure phosphorus extractability as affected by aluminum- and iron by-products and aerobic composting.

Shifts in manure phosphorus (P) chemical forms and pool sizes induced by water treatment residuals and industrial mineral by-products are largely undefined. We conducted a manure P fractionation study to determine mechanisms of reduction of dissolved reactive phosphorus (DRP) in poultry manure upon mineral by-product additions. The effects of composting on the P immobilization efficacy of the by-products were determined using laboratory self-heating composting simulators. The mineral by-products included an aluminum-water treatment residual (Al-WTR) and an iron-rich titanium-processing by-product. The noncomposted manure averaged 0.11 g g(-1) of total P as DRP forms. The by-products significantly reduced manure DRP, by an average of 39 and 48% in the Al- and the Fe-treated manure, respectively. The by-products also reduced the 0.5 M NH4F-extractable phosphorus (FEP) fraction. Shifts in P forms between FEP and 0.1 M NaOH-extractable phosphorus (SHEP) depended upon the Al and Fe contents of the by-products while the combined FEP + SHEP pool remained constant. Phosphate sorption measurements supported the observations that the Fe-rich by-product was more effective at reducing manure DRP and enhancing the formation of SHEP forms at the expense of FEP than the Al-WTR. Composting had no effect on the efficacy of either by-product to reduce DRP. Potential mechanisms of enhanced P stabilization in treated manure upon composting included chemical shifts from the DRP and FEP fractions to the citrate-bicarbonate-dithionite extractable P fraction. Thus, the choice of P immobilization agents affected the stability of immobilized P forms and should be taken into consideration in developing manure processing and nutrient stabilization methods.

Animals↗

Polycyclic aromatic hydrocarbons and polychlorobiphenyls in wastewaters and sewage sludges from the Paris area (France).

Polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs) were investigated during 1999-2000 in the sewerage system to the Seine Aval treatment plant which drains the Paris area (France). Contamination of sludges from storage chambers indicated a local origin typical of motor car traffic with PAH concentrations (Sigma16) ranging from 14 to 31 mg kg(-1) of dry weight, pyrene/fluoranthene ratios ranging between 1.12 and 1.48 and PCB concentrations (Sigma7) ranging from 0.07 to 0.65 mg kg(-1) of dry weight. In the same way, sludges from sand removal tanks displayed PAH values related to the importance of traffic whereas PCBs were rather characteristic of a diffuse origin. Among the five sewers entering the Seine Aval treatment plant, Clichy Argenteuil, which drains a large industrial area, was the most polluted one (PAHs as Sigma3, 10 kg year(-1); PCBs as Sigma7, 3.5 kg year(-1)). Seasonal variations were observed for PAHs with higher levels in winter in relation with the occurrence of combustion processes. Ninety-eight percent of the PAHs and 76% of the PCBs were extracted during the treatment plant processes, the primary clarifier of which retained 50% of the pollutants. Fluoranthene and PCB concentrations in the final dehydrated sludge (mean values) were 1.07 and 0.623 mg kg(-1) of dry weight, respectively. For PCBs, a significant correlation was found (r=0.668, P<0.001) between wastewater and dehydrated sludge concentrations. Yearly measured amounts of PAHs (Sigma3) were twice higher in sludges (76.3 kg) than in precipitations (37.7 kg) whereas those of PCBs (Sigma7) brought by total atmospheric fallout (17.6 kg) and sludges (21.1 kg) did not differ significantly.

Chromatography, Gas↗