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Isolation and process parameter optimization of Aspergillus sp. for removal of chromium from tannery effluent.

Five morphologically different fungi were isolated from leather tanning effluent in which Aspergillus sp. and Hirsutella sp. had higher potential to remove chromium. The potential of Aspergillus sp. for removal of chromium was evaluated in shake flask culture in different pH, temperature, inoculums size, carbon and nitrogen source. The maximum chromium was removed at pH 6, temperature 30 degrees C, sodium acetate (0.2%) and yeast extract (0.1%). Aspergillus sp. was applied in 2l bioreactor for removal of chromium, and it was observed that 70% chromium was removed after 3 days.

Aspergillus↗

Respiratory function and immunological status in cocoa and flour processing workers.

Respiratory function and immunological status were studied in 40 cocoa and 53 flour processing workers employed as packers in a confectionery industry and in 65 unexposed control workers in the same industry. A high prevalence of chronic respiratory symptoms was recorded in exposed workers, varying from 5.0% to 30.0% in cocoa workers and from 5.7% to 28.3% in flour workers. Occupational asthma was diagnosed in 2 (5%) of the cocoa workers and in 3 (5.7%) of the flour workers. None of the control workers suffered from occupational asthma. The prevalence of almost all chronic respiratory symptoms was significantly greater in cocoa and flour workers than in control workers. There was also a high prevalence of acute symptoms that developed during the work shift, being highest for cough (cocoa: 57.5%; flour: 50.9%) and eye irritation (cocoa: 50.0%; flour: 54.7%). Significant across-shift reductions of ventilatory capacity were recorded in exposed workers, being largest for flow rates at 50% and the last 25% of the vital capacity on maximum expiratory flow-volume (MEFV) curves (FEF50, FEF75). The prevalence of positive skin tests for cocoa (60.2%) was significantly higher than the prevalence of positive skin tests for flour (25.8%) among the 93 exposed workers (p < 0.05). Control workers had significantly lower prevalences of positive skin tests to cocoa (4.6%) and flour (12.3%) than exposed workers (p < 0.01). Increased total serum IgE levels were found in 17.5% of cocoa and in 18.7% of flour workers; none of the control workers had increased IgE levels. Bronchoprovocation testing demonstrated significant decreases in lung function following inhalation of cocoa dust extract and flour dust in workers with respiratory symptoms and large across-shift reductions in lung function. Dust concentrations in the working environment were higher than those recommended by Croatian standards. These data suggest that workers employed in the processing of cocoa and flour may be at a high risk for the development of allergic sensitization and respiratory impairment.

Adult↗

A novel process utilizing subcritical water and nitrilotriacetic acid to extract hazardous elements from MSW incinerator fly ash.

An effective process for hazardous element extraction from municipal solid waste incinerator fly ash was developed. The key trait of the process was to extract most of the hazardous elements out of the ash but leave Ca and Si inside the residue. In the extraction process, the ash was firstly pre-washed with water, then subjected to subcritical water (SC water) treatment and nitrilotriacetic acid (NTA) extraction. SEM images indicated that SC water is strong enough to destroy the ash particles, thus greatly improving hazardous elements extraction efficiency in the subsequent NTA extraction process. The extraction percentages for Cr, As, Se, Cd and Pb under SC water+NTA treatment were around 2-6 times higher than those treated by NTA at room conditions. The preferable SC treatment temperature was 573 K and the treatment time was 3 h. The optimum NTA concentration, vibration time and liquid/solid ratio were 0.8 M, 5 h and 10:1 (ml/g), respectively. Furthermore, it was found that introduction of a suitable amount of sulfuric acid into the extraction solution could extract more than 90% of most of the hazardous elements out of the ash.

Carbon↗

Temporal and spatial variations of airborne Mg, Cl, Na, Ca and K in rural areas of Norway

The Norwegian air chemistry monitoring network includes measurements of gaseous and particulate compounds in air at 12 rural sites. The sampling method is designed to determine concentration levels of sulphur and nitrogen compounds and uses a three-stage filter pack sampler. Concentration levels of base-cations and sea salts may also be determined from the chemical analysis of the filter extracts. In this study, concentration levels of water soluble Mg, Cl, Na, Ca and K in air have been investigated in terms of temporal, seasonal and spatial trends during the period 1986-1996. The results have been combined with precipitation chemistry data to estimate total deposition, and to evaluate the relative importance of dry deposition. While Na, Cl and Mg are of marine origin, which is clearly reflected in concentration ratios and spatial gradients, Ca and K are of a mixed origin. Air trajectory analyses indicate that air masses originating in Eastern Europe result in significantly higher airborne concentrations of non-sea salt Ca and K, than air from other sectors. Major sources of airborne Ca and K in Norway are expected to be of anthropogenic origin from combustion plants and industrial processes in Eastern Europe, while aeolian dust from agricultural areas within Europe or from Sahara is of limited importance. Concentration levels of non-sea salt base-cations in air were 20-30% lower during the 1990s, compared to 1986-1989. Precipitation chemistry data available since the early 1980s indicate even larger reductions, and in the order of 50%. Total deposition of non-sea salt base-cations during 1993-1996 was generally below 10 mmol(c) m-2 year-1, whereas in coastal areas with large precipitation amounts, inputs exceeding 15 mmol(c) m-2 year-1 occurred. Deposition of base-cations may thus counteract up to 25% of the strong acid anion input, but more typically in the order of 5-10%. Dry deposition of non-sea salt base-cations of southern Norway is comparatively small, typically contributing 10-30% to the total deposition in the inland areas, and relatively less in other regions.

Journal Article↗

Used lubricating oil recycling using hydrocarbon solvents.

A solvent extraction process using new hydrocarbon solvents was employed to treat used lubricant oil. The solvents used were liquefied petroleum gas (LPG) condensate and stabilized condensate. A demulsifier was used to enhance the treatment process. The extraction process using stabilized condensate demonstrated characteristics that make it competitive with existing used oil treatment technologies. The process is able to reduce the asphaltene content of the treated lubricating oil to 0.106% (w/w), the ash content to 0.108%, and the carbon residue to 0.315% with very low levels of contaminant metals. The overall yield of oil is 79%. The treated used oil can be recycled as base lubricating oil. The major disadvantage of this work is the high temperature of solvent recovery. Experimental work and results are presented in detail.

Carbon↗

The affinity concept in bioseparation: evolving paradigms and expanding range of applications.

The meaning of the word affinity in the context of protein separation has undergone evolutionary changes over the years. The exploitation of molecular recognition phenomenon is no longer limited to affinity chromatography modes. Affinity based separations today include precipitation, membrane based purification and two-phase/three-phase extractions. Apart from the affinity ligands, which have biological relationship (in vivo) with the target protein, a variety of other ligands are now used in the affinity based separations. These include dyes, chelated metal ions, peptides obtained by phage display technology, combinatorial synthesis, ribosome display methods and by systematic evolution of ligands by exponential enrichment (SELEX). Molecular modeling techniques have also facilitated the designing of biomimetic ligands. Fusion proteins obtained by recombinatorial methods have emerged as a powerful approach in bioseparation. Overexpression in E. coli often result in inactive and insoluble inclusion bodies. A number of interesting approaches are used for simultaneous refolding and purification in such cases. Proteomics also needs affinity chromatography to reduce the complexity of the system before analysis by electrophoresis and mass spectrometry are made. At industrial level, validation, biosafety and process hygiene are also important aspects. This overview looks at these evolving paradigms and various strategies which utilize affinity phenomenon for protein separations.

Affinity Labels↗

Downstream processing in marine biotechnology.

Downstream processing is one of the most underestimated steps in bioprocesses and this is not only the case in marine biotechnology. However, it is well known, especially in the pharmaceutical industry, that downstreaming is the most expensive and unfortunately the most ineffective part of a bioprocess. Thus, one might assume that new developments are widely described in the literature. Unfortunately this is not the case. Only a few working groups focus on new and more effective procedures to separate products from marine organisms. A major characteristic of marine biotechnology is the wide variety of products. Due to this variety a broad spectrum of separation techniques must be applied. In this chapter we will give an overview of existing general techniques for downstream processing which are suitable for marine bioprocesses, with some examples focussing on special products such as proteins (enzymes), polysaccharides, polyunsaturated fatty acids and other low molecular weight products. The application of a new membrane adsorber is described as well as the use of solvent extraction in marine biotechnology.

Aquaculture↗

Kinetics of tropomyosin denaturation as a predictive model for verifying thermal processing of beef products.

An enzyme-linked immunosorbent assay (ELISA) was developed to study thermal denaturation of tropomyosin (Tm) using the time-temperature requirements for cooked beef. The ELISA employed a monoclonal antibody (MAb 2C9) raised against bovine Tm for quantifying residual Tm in muscle extracts. The specificity of MAb 2C9 to bovine Tm was demonstrated by Western blot and the analytical validity of ELISA was confirmed by dot blot. Thermal denaturation of Tm, in the temperature range between 54.4 and 70.0 degrees C, showed first-order dependency. Kinetic parameters of Tm denaturation were derived from isothermal heating of beef muscle extract at 54.4, 57.2, 60.0, and 62.8 degrees C. Temperature dependency of the rate constant (k) was demonstrated by Arrhenius plot; the activation energy (E(a)) of Tm denaturation was determined to be 484 kJ x mol(-1). A mathematic model describing the impact of the heating time-temperature on Tm denaturation was developed. Predicted Tm from the integrated time-temperature model agreed closely with the measured Tm in dynamically heat-processed beef samples. Percent errors between the measured and the predicted values ranged from -5.1 to 5.3%. The kinetic model provides an accurate and reproducible prediction of the impact of actual heating time-temperature on residual Tm in cooked beef. The MAb-based ELISA and kinetic model developed in this study have the potential to be adapted by the meat industry as a quality control tool.

Animals↗

Atmospheric xenon radioactive isotope monitoring.

The Comprehensive Nuclear Test Ban Treaty (CTBT) organisation is implementing a world-wide monitoring network in order to check that the State Signatories comply with the treaty. One of the monitoring facilities consists of an atmospheric noble gas monitoring equipment. According to the requirements annexed in the treaty, the French Atomic Energy Commission (CEA) developed a device, called SPALAX, which automatically extracts xenon from ambient air and makes in situ measurements of the activities of four xenon radioisotopes (131mXe, 133mXe, 133Xe, 135Xe). The originality of this device is noticeable essentially in the gas sample processing method: thanks to the coupling of a gas permeator and of a noble gas specific adsorbent, it can selectively extract and concentrate xenon to more than 3 x 10 E6. This process is carried out continuously without cryogenic cooling, without any regeneration time. The detection of the xenon radioactive isotopes is done automatically by high spectral resolution gamma spectrometry, a robust technology well-suited for on-field instrumentation. In the year 2000, a prototype was involved in an international evaluation exercise directed by the CTBT organisation (CTBTO). This exercise demonstrated that the SPALAX equipment perfectly met the requirements of the CTBTO for such systems. On the basis of the continuous 24-h resolution record of the atmospheric xenon radioactive isotopes concentrations, the SPALAX system also demonstrated that ambient levels of 133Xe can fluctuate quickly from less than the detection limit to over 40 x 10(-3) Bq m(-3). In order to build an industrial version of this equipment, the CEA entered into a partnership with a French engineering company (S.F.I., Marseille, France), which is now able to produce an industrial version of SPALAX, i.e. more compact and more efficient than the prototypes. The 133Xe minimum detectable concentration is 0.15 x 10(-3) Bq m(-3) air per 24 h sampling cycle.

Air Pollutants, Radioactive↗

Value-added food: single cell protein.

The alarming rate of population growth has increased the demand for food production in third-world countries leading to a yawning gap in demand and supply. This has led to an increase in the number of hungry and chronically malnourished people. This situation has created a demand for the formulation of innovative and alternative proteinaceous food sources. Single cell protein (SCP) production is a major step in this direction. SCP is the protein extracted from cultivated microbial biomass. It can be used for protein supplementation of a staple diet by replacing costly conventional sources like soymeal and fishmeal to alleviate the problem of protein scarcity. Moreover, bioconversion of agricultural and industrial wastes to protein-rich food and fodder stocks has an additional benefit of making the final product cheaper. This would also offset the negative cost value of wastes used as substrate to yield SCP. Further, it would make food production less dependent upon land and relieve the pressure on agriculture. This article reviews diversified aspects of SCP as an alternative protein-supplementing source. Various potential strains and substrates that could be utilized for SCP production are described. Nutritive value and removal of nucleic acids and toxins from SCP as a protein-supplementing source are discussed. New processes need to be exploited to improve yield. In that direction the solid state fermentation (SSF) method and its advantages for SCP production are highlighted.

Journal Article↗

Separation of genomic DNA, RNA, and open circular plasmid DNA from supercoiled plasmid DNA by combining denaturation, selective renaturation and aqueous two-phase extraction.

In the current study we developed a process for the capture of pDNA exploiting the ability of aqueous two-phase systems to differentiate between different forms of DNA. In these systems scpDNA exhibits a near quantitative partitioning in the salt-rich bottom phase. The successive recovery from the salt rich bottom phase is accomplished by a novel membrane step. The polish operation to meet final purity demands is again based on a system exploiting a combination of the denaturation of the nucleic acids present, specific renaturation of scpDNA, and an ATP system able to differentiate between the renatured scpDNA and the denatured contaminants such as ocpDNA and genomic host DNA. This polish step thus allows a rapid and efficient separation of scpDNA from contaminating nucleic acids which up to date otherwise only can be accomplished with much more cumbersome chromatographic methods. In a benchmark comparison, it could be shown that the newly developed process exhibits a comparable yield to an industrial standard process while at the same time showing superior performance in terms of purity and process time. Additionally it could be shown that the developed polish procedure can be applied as a standalone module to support already existing processes.

Chemical Fractionation↗

Fermentative capacity in high-cell-density fed-batch cultures of baker's yeast.

High-cell-density fed-batch processes for bakers' yeast production will involve a low-average-specific growth rate due to the limited oxygen-transfer capacity of industrial bioreactors. The relationship between specific growth rate and fermentative capacity was investigated in aerobic, sucrose-limited fed-batch cultures of an industrial bakers' yeast strain. Using a defined mineral medium, biomass concentrations of 130 g dry weight/L were reproducibly attained. After an initial exponential-feed phase (mu = 0.18 h(-1)), oxygen-transfer limitation necessitated a gradual decrease of the specific growth rate to ca. 0.01 h(-1). Throughout fed-batch cultivation, sugar metabolism was fully respiratory, with a biomass yield of 0.5 g biomass/g sucrose(-1). Fermentative capacity (assayed off-line as ethanol production rate under anaerobic conditions with excess glucose) showed a strong positive correlation with specific growth rate. The fermentative capacity observed at the end of the process (mu = 0.01 h(-1)) was only half that observed during the exponential-feed phase (mu = 0.18 h(-1)). During fed-batch cultivation, activities of glycolytic enzymes, pyruvate decarboxylase and alcohol dehydrogenase in cell extracts did not exhibit marked changes. This suggests that changes of fermentative capacity during fed-batch cultivation were not primarily caused by regulation of the synthesis of glycolytic enzymes.

Alcohol Dehydrogenase↗

Bioconversion of phytosterols to androstanes by mycobacteria growing on sugar cane mud.

Direct sterol conversion of sugar cane mud (residue) by Mycobacterium sp. was demonstrated to be possible technologically, thus avoiding sugar cane oil extraction and further processes of extraction and purification of phytosterols from this oil. Indeed, mycobacterial cells were able to convert phytosterols from sugar cane mud into 4-androstene-dione (AD) and 1,4 androsta-diene-3,17-dione (ADD). For the various concentrations assayed, concomitant higher yields for both androstanes were achieved at 20% (w/w) sugar cane mud in media. Furthermore, conversions were similar to those from other substrates, such as a mixture of phytosterols. The results suggest that the mycobacterial cell is able to easily access and bioconvert sugar cane mud phytosterols.

Androstanes↗

Determination of organic matter in grinding sludges by UV spectrophotometry.

The aim of this paper is to determinate the total organic matter content of grinding sludges by a simple and fast methodology based on a solvent extraction followed by a UV spectrophotometry analysis. Several UV spectra exploitation methods are used. The multiwavelength deconvolution procedure provides quantitative results in good accordance with those obtained by a gravimetric method, possibly underestimated and enables a separate quantification of the cuttings fluids and the lubricants in the sludges. The normalisation procedure and the search of isobestic points provide important qualitative information on the evolution and on the composition of the oils used in metal processing. Therefore, this study is a first step for the development of a tool for the characterisation of organic matter in grinding sludges.

Hydrocarbons↗

The evaluation of mixtures of yeast and potato extracts in growth media for biomass production of lactic cultures.

The effectiveness of yeast extracts (YE) and potato extracts (PE) to promote growth of seven lactic cultures was evaluated by automated spectrophotometry (AS). Two aspects of the growth curve were analysed: (1) maximum biomass obtained (using ODmax) and (2) highest specific growth rate mu(max)) Eleven lots from the same PE-manufacturing process were examined for lot-to-lot variability. The ODmax values of three of the seven strains were significantly affected by lot source, but mu(max) was not significantly affected. The growth of bacteria was systematically lower in base medium containing 100% PE than in base medium containing 100% YE for both ODmax or mu(max) data, which could be related to the lower content in nitrogen-based compounds in PE. In AS assays, highest OD values for Lactobacillus casei EQ28, Lactobacillus rhamnosus R-011, Lactobacillus plantarum EQ12, and Streptococcus thermophilus R-083 were obtained with a mixture of PE and YE. Fermentations (2 L) were also carried out to determine the accuracy of AS to predict biomass levels obtained under fermentation trials. In these fermentations, replacement of 50% YE with PE was shown to enable good growth of S. thermophilus. With L. rhamnosus R-011, a high correlation (R2 = 0.95) was found between ODmax data obtained in the AS assays and that of the 2-L bioreactor when the same growth medium was used for both series of fermentations. However, AS was not as efficient when industrial media were used for the bioreactor assays. The relationship was still good for ODmax between AS data and that of the bioreactor data with L. rhamnosus R-011 in industrial LBS medium (R2 = 0.87), but was very poor with the S. thermophilus R-083 on Rosell #43 industrial medium (R2 = 0.33). Since PE cost 40% less than YE, there are strong economic advantages in considering such a partial replacement of YE by PE.

Bacteriological Techniques↗

Multi-sensor integration for on-line tool wear estimation through radial basis function networks and fuzzy neural network.

On-line tool wear estimation plays a very critical role in industry automation for higher productivity and product quality. In addition, appropriate and timely decision for tool change is significantly required in the machining systems. Thus, this paper is dedicated to develop an estimation system through integration of two promising technologies, artificial neural networks (ANN) and fuzzy logic. An on-line estimation system consisting of five components: (1) data collection; (2) feature extraction; (3) pattern recognition; (4) multi-sensor integration; and (5) tool/work distance compensation for tool flank wear, is proposed herein. For each sensor, a radial basis function (RBF) network is employed to recognize the extracted features. Thereafter, the decisions from multiple sensors are integrated through a proposed fuzzy neural network (FNN) model. Such a model is self-organizing and self-adjusting, and is able to learn from the experience. Physical experiments for the metal cutting process are implemented to evaluate the proposed system. The results show that the proposed system can significantly increase the accuracy of the product profile.

Journal Article↗

Isolation of Deinococcus species from commercial oyster extract.

Deinococci with radiation resistance greater than that of Deinococcus radiophilus (ATCC 27603) were isolated from three commercial oyster extracts stored at 4, 20, and 30 degreesC. During storage the number of other bacteria declined and deinococci became the predominant group in the microflora, particularly at 20 degreesC, although at 30 degreesC the number of deinococci as well rapidly declined. The results suggest that the natural habitat of deinococci is an aerobic environment containing a slightly elevated saline content, soluble protein, and low sugar levels.

Animals↗

Transgenic plants as factories for biopharmaceuticals.

Plants have considerable potential for the production of biopharmaceutical proteins and peptides because they are easily transformed and provide a cheap source of protein. Several biotechnology companies are now actively developing, field testing, and patenting plant expression systems, while clinical trials are proceeding on the first biopharmaceuticals derived from them. One transgenic plant-derived biopharmaceutical, hirudin, is now being commercially produced in Canada for the first time. Product purification is potentially an expensive process, and various methods are currently being developed to overcome this problem, including oleosin-fusion technology, which allows extraction with oil bodies. In some cases, delivery of a biopharmaceutical product by direct ingestion of the modified plant potentially removes the need for purification. Such biopharmaceuticals and edible vaccines can be stored and distributed as seeds, tubers, or fruits, making immunization programs in developing countries cheaper and potentially easier to administer. Some of the most expensive biopharmaceuticals of restricted availability, such as glucocerebrosidase, could become much cheaper and more plentiful through production in transgenic plants.

Antibodies↗