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Ethanol production from lignocellulosic byproducts of olive oil extraction.

The recent implementation of a new two-step centrifugation process for extracting olive oil in Spain has substantially reduced water consumption, thereby eliminating oil mill wastewater. However, a new high sugar content residue is still generated. In this work the two fractions present in the residue (olive pulp and fragmented stones) were assayed as substrate for ethanol production by the simultaneous saccharification and fermentation (SSF) process. Pretreatment of fragmented olive stones by sulfuric acid-catalyzed steam explosion was the most effective treatment for increasing enzymatic digestibility; however, a pretreatment step was not necessary to bioconvert the olive pulp into ethanol. The olive pulp and fragmented olive stones were tested by the SSF process using a fed-batch procedure. By adding the pulp three times at 24-h intervals, 76% of the theoretical SSF yield was obtained. Experiments with fed-batch pretreated olive stones provided SSF yields significantly lower than those obtained at standard SSF procedure. The preferred SSF conditions to obtain ethanol from olives stones (61% of theoretical yield) were 10% substrate and addition of cellulases at 15 filter paper units/g of substrate.

Biodegradation, Environmental↗

Chemical evaluation of waste from Thaumatococcus danielli (Benth) processing.

Three different batches of Thaumatococcus danielli (Benth) fruits were collected at different fruiting seasons. The proximate compositions of the pericarp and seeds were determined on a dry weight basis. Partial characterization of the lipid extract was carried out and tannin content determined. The average moisture, ash and lipid contents of the pericarp were higher (89.2+/-4.17%, 20.7+/-1.10% and 11.6+/-1.23%, respectively) than those of the seed (31.2+/-4.66%, 9.1+/-0.71% and 8.2+/-1.64%, respectively). Crude protein and total carbohydrate were, however, higher in the seed (9.5+/-4.38% and 69.4+/-11.52%, respectively) than in the pericarp (4.5+/-2.21% and 6.3+/-3.94%, respectively). The dietary fiber contents of both pericarp and seed were high. Starch content was very low in the pericarp (0.4+/-0.0%) compared to the seed (66.28+/-9.21%). Tannin content in the pericarp and seeds was 12.1+/-1.52 mg/g and 21.9+/-2.28 mg/g, respectively. The oil extracted from both pericarp and seeds was light-yellow in color and slightly solid at room temperature. Unsaturation level of T. danielli oil was low, while free fatty acids were high. The results obtained suggested that T. danielli waste could potentially be a raw material in livestock feed formulation.

Analysis of Variance↗

Comparative study of the smooth muscle contractor activity of airborne dusts and of dustiness in cotton, flax, and jute mills.

A bioassay technique using isolated guinea-pig ileum was employed to compare the smooth muscle contractor activity of various dusts from mills in which the prevalence of byssinosis was known. The activity of dust from a mill spinning a coarse grade of cotton was several times greater than that in dust from a mill processing a fine grade of cotton. There was a similar order in the difference of the prevalence of byssinosis in these mills. However, the activities of fine cotton, flax, and jute dusts were very similar to each other, in spite of marked differences in the prevalence of byssinosis in these mills. For cotton dust, smooth muscle contractor activity was associated with all particle sizes, although the lowest level of activity was found in the largest sized fraction (less than 2 mm). Activity in the cotton dust extracts was not correlated with nitrogen, carbohydrate, or potassium content. However, about one-fifth of the activity of a cotton dust extract was associated with an insoluble particulate fraction. The possible chemical nature of the water-soluble contractor agent is discussed. It is concluded that, until the role of this agent in the pathogenesis of byssinosis has been established, the bioassay technique cannot be employed as a means of assessing the byssinogenic potential of cotton dust.

Air Pollutants↗

Toxicology of cupric salts on honeybees. V. Gluconate and sulfate action on gut alkaline and acid phosphatases.

Some aspects of putative nontarget effects of cupric ions systemically fed to honeybees against their parasite mite Varroa jacobsoni have been investigated on the host phosphatases. The alkaline and acid forms extracted from the guts of worker bees exhibited substrate-inhibition features. Upon detailed kinetic analysis, cupric organic salts indicate activation effects at concentrations of about 1 mM. Concentrations up to 10 mM (alkaline form) and 25 mM (acid form) induced no important changes, except a partial quenching of the substrate-inhibition process, characterized by a wide increase in the constant of apparent inhibitory binding of substrate to the enzyme-substrate complex. Partial purification gave a single alkaline form with quite similar kinetic behavior in the absence of natural ions as in crude extracts. Cupric gluconate and sulfate demonstrated similar patterns, except an increase of the apparent Hill coefficient by sulfate only. The substrate constant of acid phosphatases was decreased at high cupric gluconate doses while its maximum velocity was biphasically increased (with observed maximum at 1 mM), resulting in a sustained activation. Chemiluminescence studies revealed that cupric ion activation is counteracted by oxygen radicals generated by cupric ions and also, in vitro, by the artificial substrate para-nitrophenylphosphate. The para-nitrophenol molecules released from the reaction are therefore responsible for biphasic effects selectively observed with gluconate salts. In apicultural practice, neither blockade of activity nor dramatic changes are to be expected at doses administered to bees against the parasite.

Acid Phosphatase↗

Influence of corn steep liquor and glucose on colonization of control and CCB (Cu/Cr/B)-treated wood by brown rot fungi.

There are increasing problems with regard to the disposal of treated wood waste. Due to heavy metals or arsenic in impregnated wood waste, burning and landfill disposal options are not considered to be environmentally friendly solutions for dealing with this problem. Extraction of the heavy metals and recycling of the preservatives from the wood waste is a much more promising and environmentally friendly solution. In order to study the scale up of this process, copper/chromium/boron-treated wood specimens were exposed to copper tolerant (Antrodia vaillantii and Leucogyrophana pinastri) and copper sensitive wood decay fungi (Gloeophyllum trabeum and Poria monticola). Afterwards, the ability of fungal hyphae to penetrate and overgrow the wood specimens was investigated. The fungal growths were stimulated by immersing the specimens into aqueous solution of glucose or corn steep liquor prior to exposure to the fungi. The fastest colonization of the impregnated wood was by the copper tolerant A. vaillantii. Addition of glucose onto the surface of the wood specimens increased the fungi colonization of the specimens; however, immersion of the specimens into the solution of corn steep liquor did not have the same positive influence. These results are important in elucidating copper toxicity in wood decay fungi and for using these fungi for bioremediation of treated wood wastes.

Basidiomycota↗

The effects of treatment with enzymatic preparations and tannin phenolic absorbent agents in extracting the olive oil by single pressure process on the oil yield, on the analytical characteristics of the oils of the vegetation waters and of the waste waters.

The vegetation waters of the must, the oils and the waste waters extracted from olive pastes, treated by enzymatic preparations (Cellulase, Hemicellulase, Polygalacturonase and Protease) and tannin phenolic adsorbent agents (Polyclar AT, Methylcellulose, Albumine) employed during crushing of olive were analysed:--the vegetation waters in order to justify some technological results (oil yields and extraction speed) obtained by the treatments;--the oils in order to examine the effects of the treatments on the same chemical characteristics and the stability to rancidity. --The waste waters to verify the possible influence of the treatments on the BOD. From the results relative to the vegetation waters, it has been suggested that the increase of the "detached oil" does occur by two different biochemical mechanisms which involve the constitutive lipoprotein of the oil drop membrane and colloidal constituents of the liquid and solid phase of the pastes. For what concerns the oils, the treatments induce the slight modification of the fatty acid composition and chromatic characteristics, a higher stability to rancidity, a qualitative and quantitative increase of the "headspace" violates constituents above oils. The waste waters obtained from the olive pastes show lower BOD5 values.

Adsorption↗

Batch fermentations on synthetic mixed sugar and starch medium with amylolytic lactic acid bacteria.

The green crop drying industry in Denmark uses Italian rye grass, clover, and alfalfa as raw materials for the production of green pellets. The green crop drying industry solves its energy economical problems by heating and pressing of the green crop before drying. The produced sidestream is called brown juice. Brown juice was shown to be an excellent medium for lactic acid fermentation. The aim of this study was to investigate the utilisation of brown juice in the production of polylactic acid, where wheat starch would be added to increase the lactic acid yield and, thus, the feasibility of the process. A number of amylolytic lactic acid bacteria have been identified, and in this work, six different strains were tested for their ability to produce alpha-amylase and to utilise all sugars with high lactic acid yield in a medium with a complex composition of free sugars (brown juice) and starch. Lactobacillus plantarum A6 was the only strain that showed both a good lactic acid production and utilisation of starch in this medium. The growth rate of this strain was approximately 0.4 h(-1) and the lactic acid yield was 0.7.

Carbohydrate Metabolism↗

Activated carbon adsorption of PAHs from vegetable oil used in soil remediation.

Vegetable oil has been proven to be advantageous as a non-toxic, cost-effective and biodegradable solvent to extract polycyclic aromatic hydrocarbons (PAHs) from contaminated soils for remediation purposes. The resulting vegetable oil contained PAHs and therefore required a method for subsequent removal of extracted PAHs and reuse of the oil in remediation processes. In this paper, activated carbon adsorption of PAHs from vegetable oil used in soil remediation was assessed to ascertain PAH contaminated oil regeneration. Vegetable oils, originating from lab scale remediation, with different PAH concentrations were examined to study the adsorption of PAHs on activated carbon. Batch adsorption tests were performed by shaking oil-activated carbon mixtures in flasks. Equilibrium data were fitted with the Langmuir and Freundlich isothermal models. Studies were also carried out using columns packed with activated carbon. In addition, the effects of initial PAH concentration and activated carbon dosage on sorption capacities were investigated. Results clearly revealed the effectiveness of using activated carbon as an adsorbent to remove PAHs from the vegetable oil. Adsorption equilibrium of PAHs on activated carbon from the vegetable oil was successfully evaluated by the Langmuir and Freundlich isotherms. The initial PAH concentrations and carbon dosage affected adsorption significantly. The results indicate that the reuse of vegetable oil was feasible.

Adsorption↗

Validated sandwich enzyme-linked immunosorbent assay for casein and its application to retail and milk-allergic complaint foods.

Cows' milk is a commonly allergenic food. Cross-contamination of milk proteins into nondairy, kosher-pareve foods prepared on shared processing equipment can cause severe, life-threatening reactions in milk-allergic individuals. A sandwich-type enzyme-linked immunosorbent assay (ELISA; 96-well plate format) was developed for the detection of undeclared casein in foods. Rabbit anti-casein antibodies were used as the capture reagent. Food samples and standards were ground, extracted in 0.01 M phosphate-buffered saline, clarified by centrifugation, and added to the wells. Goat anti-casein antibodies were employed as the detector antibody, and the amount of antibody bound was determined with a commercial rabbit anti-goat immunoglobulin conjugated to alkaline phosphatase, with subsequent substrate reaction. Antibodies developed were specific to casein, with no cross-reaction observed with 30 foods and food ingredients. Non-milk-containing products such as fruit juices, fruit juice bars, sorbets, and dark and pareve-labeled chocolate were purchased from June 2002 through June 2003. In addition, samples allegedly causing eight milk-allergic consumer complaints were analyzed. The ELISA had a detection limit of less than 0.5 ppm of casein. The casein content in the analyzed foods ranged from less than 0.5 ppm to more than 40,000 ppm casein; undeclared casein residues were found in all of the samples implicated in allergic reactions. The levels of milk contamination in some of the other surveyed products could also be hazardous for milk-allergic consumers. This ELISA method provides a useful quality control tool for the food industry and could also be used as a validation of kosher-pareve status.

Animals↗

Motor alterations associated with exposure to manganese in the environment in Mexico.

Overexposure to manganese (Mn) causes neurotoxicity (a Parkinson-like syndrome) or psychiatric damage ("manganese madness"). Several studies have shown alterations to motor and neural behavior associated with exposure to Mn in the workplace. However, there are few studies on the effects of environmental exposure of whole populations. We studied the risk of motor alterations in people living in a mining district in Mexico. We studied 288 individual people (168 women and 120 men) from eight communities at various distances from manganese extraction or processing facilities in the district of Molango. We measured manganese concentrations in airborne particles, water, soil and crops and evaluated the possible routes of Mn exposure. We also took samples of people's blood and determined their concentrations of Mn and lead (Pb). We used "Esquema de Diagnóstico Neuropsicológico" Ardila and Ostrosky-Solís's neuropsychological battery to evaluate motor functions. Concentrations of Mn in drinking water and maize grain were less than detection limits at most sampling sites. Manganese extractable by DTPA in soils ranged between 6 and 280 mg kg(-1) and means were largest close to Mn extraction or processing facilities. Air Mn concentration ranged between 0.003 and 5.86 microg/m(3); the mean value was 0.42 microg/m(3) and median was 0.10 microg/m(3), the average value (geometric mean) resulted to be 0.13 microg/m(3). Mean blood manganese concentration was 10.16 microg/l, and geometric mean 9.44 microg/l, ranged between 5.0 and 31.0 mcrog/l. We found no association between concentrations of Mn in blood and motor tests. There was a statistically significant association between Mn concentrations in air and motor tests that assessed the coordination of two movements (OR 3.69; 95% CI 0.9, 15.13) and position changes in hand movements (OR 3.09; CI 95% 1.07, 8.92). An association with tests evaluating conflictive reactions (task that explores verbal regulations of movements) was also found (OR 2.30; CI 95% 1.00, 5.28). It seems from our results that people living close to the manganese mines and processing plants suffer from an incipient motor deficit, as a result of their inhaling manganese-rich dust.

Adult↗

Selective recovery of anthocyanins and hydroxycinnamates from a byproduct of citrus processing.

The adsorption-desorption performances of commercial resins (two polystyrene-divinylbenzene copolymers and one methacrylic) were tested in column studies for the selective recovery of anthocyanins and hydroxycinnamates from pigmented pulp wash (PW), a byproduct of blood orange juice processing. Methanol, ethanol, and their mixtures with different percentages of water as eluents were tested in order to investigate the selective desorption of these natural antioxidants with the goal of minimizing the presence of other compounds, mostly flavanones and limonoids, in the concentrated eluates. The results indicated that polystyrene-divinylbenzene resins were able to adsorb a major amount of anthocyanins and hydroxycinnamates and to desorb them in more-concentrated fractions. No selectivity was observed using pure methanol and ethanol which resulted in concentrates with no further separation of components from the starting raw PW. The ethanol/water concentrates showed higher yields than the fractions attained using methanol/water; however, a lower selectivity toward anthocyanin pigments was observed. The best performing resin, EXA-118, together with the mixture methanol/water 50:50 (v/v) as best eluent, appears to be the most suitable system to obtain highly concentrated extracts. Thus, it was chosen to perform a larger experiment, to analyze the selectivity of the removal upon an increase in elution volume.

Adsorption↗

Occupational exposure to butadiene, isoprene and chloroprene.

Workers are exposed to butadiene, isoprene and chloroprene in the manufacture of these monomers and in their use in the production of various elastomers. These include styrene butadiene rubber, polybutadiene, polyisoprene, butyl rubber and neoprene. Monomer production and extraction are done in typical closed chemical process units where low background levels of the monomers are the result of minor leaks in valves and pumps. Occasionally, higher levels occur as a result of planned or unplanned events that cause releases. Polymer production is also a closed process, but the occasional clogging of pipes and equipment with polymer requiring maintenance operations where some release is likely occurs much more often than for monomer production. For this reason, exposure levels are generally higher on polymer production units. Polymer finishing is essentially an open process, but almost all monomer should have been stripped from the polymer before finishing. Where small amounts of solvents or monomers remain in the polymer and are volatilized in finishing, they are captured by vapor control systems. As a result, exposures in finishing are typically low. Measured levels of exposure in recent years are presented. In general, modern levels of exposure are well below OSHA, ACGIH and other applicable limits. Few measurements were made prior to the 1970s, but epidemiological estimates made by modeling suggest that levels could have been quite high in the 1940s and 1950s. In these years, manual reactor cleaning was common, and pumps often leaked.

Air Pollutants, Occupational↗

Recovery of EDTA and metal precipitation from soil flushing solutions.

This work studies the effectiveness of a process proposed for the recovery of ethylenediaminetetraacetic acid (EDTA) and metal precipitation from soil flushing solutions. Two series of experimental tests were carried out on two samples of a soil artificially contaminated with copper or lead. The metals were extracted from the soil by flushing with a 0.05 M aqueous solution of EDTA sodium salt (E-Na(2)). Cu or Pb extraction efficiencies of about 95 and 98% were observed, respectively. The two extracted solutions were then treated to obtain EDTA recovery and metal precipitation from the aqueous solution. EDTA recovery was achieved in two steps. An initial evaporation treatment lead to reduce the solution volume by about 75%. This was followed by the acidification of the residue solution, which precipitated more than 93% of the used EDTA. The precipitated EDTA was removed by filtration, and was suitable for reuse after adding an alkaline agent. Metal precipitation from the filtered solution was performed using two different methods: an almost total (99.5%) Pb precipitation in alkaline conditions was achieved after complex destabilization through the sequential addition of ferric ions and sodium phosphate, while 93.6% copper precipitation was achieved with ferrous sulfate as a destabilization agent.

Chelating Agents↗

Overview of results from the WaterTox intercalibration and environmental testing phase II program: part 2, ecotoxicological evaluation of drinking water supplies.

Because of rapid population growth, industrial development, and intensified agricultural production increasing amounts of chemicals are being released into the environment, polluting receiving water bodies around the world. Given the potential health risk associated with the presence of toxicants in water sources used for drinking yet the scarcity of available data, there is a need to evaluate these waters and develop strategies to reduce and prevent their contamination. The present study examined the applicability of a battery of simple, inexpensive bioassays in environmental management and the relevance of the test results in establishing the toxicological quality of water sources and drinking water within the framework of the eight-country WaterTox Network, sponsored by the International Development Research Centre, Ottawa, Canada. Seventy-six samples were collected from surface and groundwater sources and seven samples from drinking water treatment plants. Each sample was tested with a core battery of bioassays (Daphnia magna, Hydra attenuata, and Lactuca sativa root inhibition tests) and a limited set of physical and chemical parameters. In addition, three labs included the Selenastrum capricornutum test. When no toxic effects were found with the battery, samples were concentrated 10x using a solid-phase extraction (SPE) procedure. Nonconcentrated natural water samples produced a toxic response in 24% of cases with all three core bioassays. When all bioassays are considered, the percentage of raw samples showing toxicity with at least one bioassay increased to 60%. Of seven treated drinkingwater samples, four showed toxicity with at least one bioassay, raising the possibility that treatment processes in these instances were unable to remove toxic contaminants. The Daphnia magna and Hydra attenuata tests indicated a high level of sensitivity overall. Although only three of the eight countries used S. capricornutum, it proved to be an efficient and reliable bioassay for toxicity assessment.

Animals↗

Immobilisation of beryllium in solid waste (red-mud) by fixation and vitrification.

The objective of this study was to obtain information on the immobilization of beryllium (Be) in solid waste generated in the extraction process of beryllium from its ore, Beryl. This solid waste, termed red-mud, contains oxides of iron, aluminium, calcium, magnesium and beryllium. The red-mud waste contains beryllium at levels above the permissible limit, which prevents its disposal as solid waste. The total beryllium content in the red-mud analysed showed value ranging from 0.39 to 0.59% Be The studies showed that 50% of the total beryllium in red-mud can be extracted by water by repeated leaching over a period of 45 days. The cement mix, casting into cement blocks, was subjected to leachability studies over a period of 105 days and immobilization factor (IF factor) was determined. These IF values, of the order of 102, were compared with those obtained by performing leachability study on vitrified red-mud masses produced at different temperature conditions. Direct heating of the red-mud gave the gray coloured, non-transparent vitreous mass (as 'bad glass') showed effective immobilisation factor for beryllium in red-mud of the order of 10(4).

Beryllium↗

Anaerobic treatment of natural tannin extracts in UASB reactors.

Tannin extracts are substances commonly used in leather production processes. Since most of the steps of tannery manufacturing processes are carried out in aqueous environments, the presence of these compounds in the wastewaters is important. The aim of this work is to study the feasibility of the anaerobic degradation of three natural tannin extracts in three Upflow Anaerobic Sludge Blanket (UASB) reactors, which were fed with increasing concentrations of two condensed (quebracho and wattle) and one hydrolysable tannin extract (chestnut). Concentrations of applied extracts were 100, 200, 400, 800 and 1,000 mg/l, and 5 g/l of glucose was used as cosubstrate. Reactors were operated during 210 days and their performance was evaluated from the values of total and soluble COD, total and intermediate alkalinity, volatile fatty acids, pH and UV absorption at 280 nm. COD removal efficiencies higher than 85% were achieved in all cases. However, tannin extract removal efficiencies (based on UV-280 nm absorption measurements) were significantly lower, around 20% for condensed extracts and 60% for the hydrolysable one, when the reactors operated with the highest tannin extract concentration. The operation of the reactors was stable, commonly with alkalinity ratios below 0.30. Mass balances carried out indicate that most of the COD removal efficiencies are due to the removal of the readily biodegradable organic matter (glucose), whereas the tannin extracts are hardly anaerobically biodegradable, especially condensed extracts (wattle and quebracho).

Animals↗

Screening of bacterial contamination during gelatine production by means of denaturing gradient gel electrophoresis, focussed on Bacillus and related endospore-forming genera.

AIMS: To screen for bacterial contamination during gelatine production by means of denaturing gradient gel electrophoresis (DGGE). As members of Bacillus and related genera were found to persist in the final product, this study focussed on these taxa. METHODS AND RESULTS: Template DNA was extracted from gelatine samples at five crucial points of a gelatine production process. A primer specific for Bacillus and related genera was designed and used in a selective PCR, followed by a nested DGGE-PCR targeting the V9 region of the 16S rDNA. DGGE analysis of the resulting amplicons, and sequence analysis of selected bands, showed high sequence similarities of these bands with Bacillus fumarioli, B. licheniformis, B. coagulans and Clostridium perfringens. When the selective PCR was omitted, primarily Lactobacillus bands were retrieved. CONCLUSIONS: PCR-DGGE analysis of gelatine extracts can be used for tracing and screening of bacterial contamination during gelatine production. A selective PCR, nested with DGGE-PCR, gave much more accurate information about endospore-forming contaminants than did the direct DGGE procedure alone. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of this nested DGGE-PCR protocol may provide important information about possible hazards to the final microbiological quality and/or safety of gelatine, so allowing production parameters and/or remediation procedures may be adjusted on-line.

Bacillus↗

Utilization of renewables for lactic acid fermentation.

Originally, lactic acid was produced from pure substrates like glucose. Increasingly, however, agricultural feedstocks such as grains and green biomass are also being used as raw materials for the biotechnological production of lactic acid. A high-productivity lactic acid bacterium strain was selected, process parameters were optimized for the batch fermentation on a laboratory scale, and its performance at cultivation on a barley hydrolysate medium together with different supplements was examined. The present results for the cultivation of the Lactobacillus paracasei on complex nutrient broth are in the same range as those for another strain of the same species with pure glucose, de Man, Rogosa and Sharpe medium (MRS) minerals, peptone and yeast extract. Under these conditions, this strain was able to accumulate more than 100 g lactate/L in the MRS medium. Medium optimization experiments showed that the main part of the nitrogen-containing nutrients in the medium (peptone, yeast extract) can be replaced by protein extracts from green biomass (lucerne green juice). The green juice after pressing fresh biomass contains a series of nitrogen-containing compounds and inorganic salts, which are essential for cell growth. Thus, on laboratory scale, we have demonstrated that it is possible to substitute synthetic nutrients by renewable resources like cereals and green biomass without any loss of productivity. This high biomass concentration together with the number of living cells could increase the productivity to higher levels compared to the well-adapted synthetic nutrients of MRS.

Agriculture↗