Characterization of a vitamin B12 compound from unicellular coccolithophorid alga (Pleurochrysis carterae).

A unicellular coccolithophorid alga, Pleurochrysis carterae, contained 125.4 +/- 1.2 microg of vitamin B12 per 100 g dry cell weight of the lyophilized algal cells. A vitamin B12 compound was purified from the lyophilized algal cells and partially characterized. The silica gel 60 TLC and reversed-phase HPLC patterns of the purified pink-colored compound were identical to those of authentic vitamin B12, but not those of vitamin B12 analogues inactive for humans. When 22-week-old B12-deficient rats which excreted substantial amounts of methylmalonic acid (75.5 +/- 12.3 mg/day) in urine were fed the P. carterae (10 g per kg diet)-supplemented diet for 12 d, urinary methylmalonic acid excretion (as an index of vitamin B12 deficiency) of the rats became undetectable and hepatic vitamin B12 level of the rats was significantly increased.     

Biochar derived from dairy cattle carcasses as an alternative source of phosphorus and amendment for soil acidity

Two pot experiments were conducted to evaluate biochar derived from dead dairy cattle as a mineral fertilizer, especially phosphorus (P) fertilizer, and to clarify the effect of particle size of biochar on plant growth (Zea mays L.) and P uptake. To produce the biochar, body parts of dead cattle were placed in a charring chamber and allowed to char at 450°C for 4 h. The biochar was of high pH and rich in major plant nutrients, especially P. Application of fine biochar (< 1 mm) increased P uptake by the corn plants grown in soil of low available P status. As a result, plant growth was improved following biochar application and dry matter production was also increased. The effect of the biochar application on the P uptake and plant growth was promoted by the application of mineral nitrogen (N) fertilizer. Soil analysis after harvest indicated that the biochar application increased soil pH, available P and exchangeable calcium (Ca) and magnesium (Mg) compared with the soil before seeding, while soil available N and exchangeable potassium (K) were considerably decreased. The decrease in the soil available N was incomprehensible, because the result of the mass balance given by the difference between input as the applied N from the biochar and fertilizer N and output as the N uptake by the plants was positive. We observed a similar result in the mass balance of K to the case of N. The medium (2–4 mm) and coarse (> 4 mm) grade biochar did not significantly affect plant growth, because P uptake was not, or was only slightly, increased by the application of these biochars. Dissolution of P from the coarser biochars was probably slower than that of the fine biochar. The lower dissolution of P from the medium and coarse biochars was supported by the lower P absorption efficiency of these biochars compared with that of the fine biochar and superphosphate. The effect of fine biochar on plant growth and P uptake was similar to that of superphosphate. We can therefore conclude that fine biochar derived from cattle carcasses is an effective source of P fertilizer and amendment for soil acidity. The N and K contents in the biochar, although relatively high, cannot be relied upon as a mineral fertilizer. Further studies are needed to assess whether the N and K contents of the biochar indicate it can be regarded as a useful fertilizer.     

Issues in assessing the validity of nutrient data obtained from a food-frequency questionnaire: folate and vitamin B12 examples

OBJECTIVE: To compare methods used to assess the validity of nutrient intake data obtained from a food-frequency questionnaire (FFQ), using folate and vitamin B12 as nutrient examples. DESIGN: Cross-sectional sample from a population cohort. SETTING: Two postcode areas west of Sydney, Australia. SUBJECTS: In total, 2895 people aged 49 years and older provided dietary data using a semi-quantitative FFQ (79% of 3654 subjects examined). The validity of the FFQ was assessed against three 4-day weighed food records (WFRs) completed by 78 people (mean age 70 years). RESULTS: Folate and vitamin B12 validity data were assessed using different methods. The Spearman ranked correlations (energy-adjusted) were 0.66 for folate and 0.38 for vitamin B12. Using the Bland-Altman method, following loge transformation, no linear trend existed between the differences and means for folate and vitamin B12. Large differences existed between the FFQ and WFR in individual cases, particularly for vitamin B12. Finally, data were divided into quintile categories for the test and reference method: 79% classified folate within one quintile, 65% classified vitamin B12 within one quintile; there was no gross misclassification for folate and only 3% misclassification for vitamin B12. CONCLUSIONS: Different methods of analysis provided different information about the validity of the FFQ. Correlation coefficients should not be used alone to assess the validity of nutrient data, but should be used in conjunction with Bland-Altman analyses. Depending on the use of the data, additional assessment of classification categories is recommended. This worked example demonstrates that absolute intakes of folate and vitamin B12 should be used with caution.     

Pseudovitamin B(12) is the predominant cobamide of an algal health food, spirulina tablets.

The vitamin B(12) concentration of an algal health food, spirulina (Spirulina sp.) tablets, was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined with the microbiological method were approximately 6-9-fold greater in the spirulina tablets than the values determined with the chemiluminescence method. Although most of the vitamin B(12) determined with the microbiological method was derived from various vitamin B(12) substitutive compounds and/or inactive vitamin B(12) analogues, the spirulina contained a small amount of vitamin B(12) active in the binding of the intrinsic factor. Two intrinsic factor active vitamin B(12) analogues (major and minor) were purified from the spirulina tablets and partially characterized. The major (83%) and minor (17%) analogues were identified as pseudovitamin B(12) and vitamin B(12), respectively, as judged from data of TLC, reversed-phase HPLC, (1)H NMR spectroscopy, ultraviolet-visible spectroscopy, and biological activity using L. leichmannii as a test organism and the binding of vitamin B(12) to the intrinsic factor.     

Evaluation of extracts and isolated fraction from Capparis spinosa L. buds as an antioxidant source

The antioxidant activity of extracts from Capparis spinosa L. buds was evaluated using different in vitro tests: ascorbate/Fe(2+)-mediated lipid peroxidation of microsomes from rat liver; bleaching of 1,1-diphenyl-2-picryl-hydrazyl radical; and autoxidation of Fe(2+) ion in the presence of bathophenanthroline disulfonate. The methanolic extract showed strong activities in all of these in vitro tests. The amount of total phenols was determined in the methanolic extract. In addition, the level of rutin was calculated as 0.39% (w/w) by HPLC analysis. Our findings indicate the following: (a) the antioxidant efficiency of the methanolic extract may be attributed to its phenolic content; and (b) the antioxidant activity of the methanolic extract was maintained after removal of glucosinolates, confirming that these compounds do not interfere with the antioxidant properties of the extract. The results obtained from this study exalt the nutritional value of the flowering buds (capers) which are widely used as a source of flavor.     

Assessment of Biogas use as an Energy Source from Anaerobic Digestion of Brewery Wastewater

Energy recovery from a crossflow ultrafiltration (UF) membraneunit employed in order to improve the performance of an anaerobic contact digester for the treatment of brewery wastewater was assessed. The performance of the pilot-scale anaerobic UF membrane system was studied for over 15 months. At steady-state conditions, an organic loading rate of 28.5 kg COD m^-3 d^-1, a hydraulic retention time of 4.2 days and overall COD and BOD removal efficiencies of 99% and almost 100% were achieved, respectively. Percent methane in biogas was found to be in a range of 67–79% with the corresponding methane yield of 0.28–0.35 m³ CH₄ kg^-1 COD_removed. The potential energy recovery from the system treating brewery wastewater at an OLR of 28.5 kg COD m^-3 d^-1 was 87 MJ d^-1 which would enable to maintain all energy requirements of the feed pump, mixing and heating of the reactor contents. In addition to this, 71% of the energy requirement for recirculating the reactor content through the membranes would also be recovered.     

Tartaric acid recovery from distilled lees and use of the residual solid as an economic nutrient for lactobacillus

The recovery of tartaric acid (TA) from distilled vinification lees coming from the white and red winemaking technology was optimized using response surface methodology and Statistica 5.0 software. The sequential treatment of dissolving TA and further calcium tartrate (CaT) precipitation could be used to recover up to 92.4% of the initial TA when distilled white lees were used. The residual lees were employed as economic nutrients for lactic acid production by Lactobacillus pentosus CECT-4023 using hemicellulosic vine shoot hydrolysates as carbon source. Distilled lees after TA extraction used as nutrients provided values of lactic acid (18.4-18.9 g/L), global volumetric productivities (0.82-0.84 g/L.h), and product yields (0.69-0.70 g/g) similar to those achieved when using the general medium for Lactobacilli (18.6 g/L, 1.11 g/L.h, and 0.62 g/g, respectively) or lees without TA extraction (16.4-17.2 g/L, 0.96-1.21 g/L.h, and 0.61-0.66 g/g, respectively). This technology not only avoids pollutant disposal but also represents a commercial source of tartaric acid and economic nutrients for biotechnological processes.     

Deoxynivalenol removal from barley intended as swine feed through the use of an abrasive pearling procedure

Samples of naturally contaminated hulled barley, with varying deoxynivalenol concentrations, were subjected to an abrasive type dehulling procedure. The remaining grain fractions were analyzed for weight remaining (%), deoxynivalenol (ppm), crude protein (%CP), neutral detergent fiber (%NDF), ash (%ASH), gross energy (GE; kcal/kg), and calculated digestible energy values (DE; kcal/kg). Following the initial 15 s of pearling, 85% of the grain mass remained. Additional pearling resulted in a linear decline of grain mass. Following 15 s of pearling, the grain contained 34% of the initial deoxynivalenol content, irrespective of the initial level of contamination. Further pearling resulted in continued significant (p < 0.05) reductions in the percent of deoxynivalenol remaining to a level of 7.9% after 120 s but with significant losses in grain mass. Pearling can serve as an effective means of reducing the deoxynivalenol content of barley, with improvements in nutrient levels. However, the need to reduce the deoxynivalenol content of contaminated barley to less than 1 ppm for swine will necessitate the removal of a significant amount of the grain mass for heavily contaminated samples.     

Gas chromatography-mass spectrometry method for the determination of free amino acids as their dimethyl-tert-butylsilyl (TBDMS) derivatives in animal source food

The suitability of a one-step derivatization procedure using N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide for the simultaneous assay of 22 free amino acids and its application for their analysis in six animal source foods (pork, dry cured ham, chicken stock, fresh cheese, ripened cheese, and dry salted sardine) by GC-MS were studied. All 22 free amino acid derivatives were correctly detected and resolved. Reproducibility (%RSD) of the method was in the range of 1.9-12.2%. Detection and quantitation limits of the analytical procedure ranged from 0.01 to 0.46 mg/100 g dry weight and from 0.02 to 1.55 mg/100 g dry weight, respectively. The calibration curves were linear within the range 0.1-15.0 mg/100 g with correlation coefficient values (R(2)) from 0.9891 to 0.9983. All analyzed food products showed free amino acid contents similar to those found in the scientific literature. The proposed GC-MS method for the determination of free amino acids in animal source food can be used in routine for both analytical and research purposes.     

The use of sodium EDTA as an extractant for determining available phosphate in soil

The use of a neutral solution of 0.0025M Na₂EDTA as an extractant in the assessment of soil phosphate available to plants is described. In the soil types used, a high correlation was found between EDTA extractable soil phosphate and plant responses, as measured by phosphate content of dry matter. The dry-matter yield of plants was also closely correlated with P_DTA values. The method provides a satisfactory estimate of the availability of phosphate to wheat plants in the soils under investigation.     

Production of triacetylhydroxytyrosol from olive mill waste waters for use as stabilized bioantioxidant

A hydroxytyrosol triacetyl derivative was very efficiently produced as a highly pure stabilized antioxidant compound by a short treatment of olive mill waste water (OMWW) organic extracts, rich in hydroxytyrosol, with an acetylating mixture composed of HClO4-SiO2 and Ac2O (Chakborti and Gulhane reaction), in mild and safe conditions. A successive single step of middle pressure liquid chromatography (MPLC) purification of the reaction product was performed, with an overall yield of 35.6%. (This process, including both the Chakborti and Gulhane reaction and the MPLC purification, is protected by an international patent under PCT/IT2005/000781.) The o-diphenol triacetyl derivative was also produced by direct reaction of hydroxytyrosol, previously purified by MPLC, with HClO4-SiO2 and Ac2O, with an overall yield of 29.5%. A further procedure for the production of the hydroxytyrosol triacetyl derivative was consistent with the direct treatment of raw OMWW with the acetylating agent and a single step of MPLC purification, with an overall yield of 27.6%. The purified natural triacetylhydroxytyrosol confirmed the same strong protective effects against the oxidative stress in human cells as the corresponding synthetic compound, likely because of the biochemical activation of the acetyl derivative into the active parent hydroxytyrosol by esterases. We therefore propose the utilization of OMWW for recovering hydroxytyrosol as a natural antioxidant in a chemically stabilized form, with a good yield, which can be potentially used as a nontoxic functional component in nutritional, pharmaceutical, and cosmetic preparations.     

The use of sewage sludge as soil amendment. The need for an ecotoxicological evaluation

Background, aim, and scope  Sewage sludge use in agriculture should be limited by the presence of metals and other persistent environmental pollutants. The present study aims to contribute for the definition of a test battery of ecotoxicological assays that allows a proper ecotoxicological characterization of sludges, providing information on their potential hazard and identified “safe” application levels. Materials and methods  Three sludges from distinct sources (urban, olive-processing, and electroplating industries) were tested using avoidance and reproduction tests with earthworms (Eisenia andrei) and springtails (Folsomia candida) and plant growth tests with turnips (Brassica rapa) and oats (Avena sativa). Different soil–sludge mixture concentrations mimicking recommended/realistic field dosages were tested. Results  Only the sludge from the electroplating industry induced an avoidance response from the earthworms (EC₅₀ = 0.4 t/ha) and collembolans (no observed effect concentration (NOEC) = 15 t/ha). This sludge was the only sludge responsible for any effect on the reproductive output of the earthworms (EC₅₀ = 7.74 t/ha). Regarding collembolans, none of the sludges tested caused any significant decrease in reproduction. In higher plant tests, the two industrial sludges were toxic, causing a decrease growth in both species. The EC₂₀ values determined for B. rapa were 20.3 and 24.2 t/ha and for A. sativa 14.7 and 16.2 t/ha for sludges from olive-processing and electroplating industries, respectively. Discussion  The metal loadings of the different test sludges could partially explain the results obtained. The toxicity of the test sludge from electroplating industry observed on the tested invertebrates and plants could be explained by the high amount of total chromium from which 22.3% was in the most toxic oxidation state—Cr(VI). However, the toxicity caused by the sludge from the olive-processing industry in the test plants could be attributed to the presence of other compounds (not measured in this study) since the metal content was not high enough to induce such an effect. The absence of toxicity showed by the urban test sludge was in agreement with its low levels of metals. Conclusions  The response of the different test organisms and end points varied according to the sludge type. The urban sludge was non-toxic whereas the sludge from the electroplating industry caused a toxic effect on almost all parameters measured (avoidance behavior of both test organisms, reproduction of earthworms, and growth of both plant species). Sludge from the olive-processing industry only caused a toxic effect on growth of both plant species. By analyzing the sensitivity of the different parameters for the most toxic sludge, it was found that avoidance and reproduction were more sensitive than plant growth, whereas plant seed germination was not sensitive at all. Recommendations and perspectives  The ecotoxicological evaluation of wastes can be used as an environmental safety control of sludge use in agriculture. A tiered approach could be adopted for this purpose, incorporating avoidance tests in the first tier (screening level) and reproduction and plant growth tests in a second tier. But more evidence aiming to define the most suitable ecotoxicological test battery for specific sludges with a different contamination profile is still needed.     

Use of beer bran as an adsorbent for the removal of organic compounds from wastewater

Beer bran was found to effectively adsorb several organic compounds, such as dichloromethane, chloroform, trichloroethylene, benzene, pretilachlor, and esprocarb. Equilibrium adsorption isotherms conformed to the Freundlich isotherm (log-log linear). Adsorption of these organic compounds by beer bran was observed in the pH range of 1-11. At equilibrium, the adsorption efficiency of beer bran for benzene, chloroform, and dichiloromethane was higher than that of activated carbon. The removal of these organic compounds by beer bran was attributed to the uptake by intracellular particles called spherosomes. The object of this work was to investigate several adsorbents for the effective removal of organic compounds from wastewater.     

Hop as an interesting source of resveratrol for brewers: optimization of the extraction and quantitative study by liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Nowadays, hop is used almost exclusively by brewers for bitterness and flavor. Although hop polyphenols have been widely studied in the past decade for their antioxidant activity in the boiling kettle, very little is known about their real impact on health. The discovery of resveratrol in hop pellets highlights the potential health-promoting effect of moderate beer consumption. Here, we have optimized a quantitative extraction procedure for resveratrol in hop pellets. Preliminary removal of hydrophobic bitter compounds with toluene and cyclohexane at room temperature allows 99% trans-resveratrol recovery by ethanol:water (75:25, v/v) solid/liquid extraction at 60 degrees C. Reverse phase liquid chromatography proves an excellent means of separating isomers. In addition, we have compared two mass spectrometry ionization methods-atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI)-in both the positive and the negative modes. On the basis of standard additions applied with the optimized extraction procedure and reverse phase high-performance liquid chromatography-APCI(+)-tandem mass spectrometry, it appears that Tomahawk hop pellets (T90, harvest 2002) contain 0.5 ppm trans-resveratrol, 2 ppm trans-piceid, no cis-resveratrol, and 0.9 ppm cis-piceid.     

The use of rate equations for a quantitative description of K desorption from soils in an external electric field (electro-ultrafiltration)

Ions can be extracted from soils by applying an external electric field to a soil suspension. When a constant field strength is employed characteristic desorption parameters can be calculated from the results, such as maximum desorbable quantity, halftime, and rate constant. The method consists of a combination of electrodialysis and ultrafiltration and is called electro-ultrafiltration (EUF). It was found that the K transfer from the central compartment A to the cathode compartment B follows a first order rate law, if compartment A contains only the solution of a strong electrolyte such as KCl. If a soil suspension is introduced into compartment A the bulk of K that is transferred to compartment B has first to be desorbed from the soil particles hence the designation as desorption for the transfer from a soil suspension. In this case a second order rate law is followed. The rate equations allow the calculation of the quantity of K involved in the desorption process and the rate with which it is released. It was found that invariably only the exchangeable K takes part in the desorption reaction. The desorption rate differs between soils and depends on clay content and the quantity of exchangeable K present in the soil. It is suggested that the desorption data may provide a means of describing the soil K status by kinetic parameters.     

Characterization of Aspen ASH,Sand and Log-Yard Waste Mixtures from an Aspen-Based Oriented Strand Board Mill for Use as an Intermediate Landfill Cover

Wood ash has a variety of uses including land application, geotechnical construction and daily and final covers for landfills. The objective of this study was to determine the relevant chemical and physical properties of selected mixtures of aspen burner-ash, log-yard waste scrapings (sandy soil plus wood waste) and a sandy soil as an intermediate landfill cover. The treatment mixtures, per weight basis, included: (1) 100% sand; (2) 80/20 log-yard waste/ash; (3) 50/50 sand/ash; (4) 40/30/30 sand/log-yard waste/ash; (5) 60/40 ash/log-yard waste; (6) 100% ash; (7) 100% log-yard waste. All treatments containing ash had a very high pH (12.8–13.3) and electrical conductivity (39–105 dS m^–1). The ash contained significant amounts of beneficial nutrients, but soil solution dominance by sodium carbonate suppressed concentrations of calcium and magnesium. Treatments containing ash showed lower bulk densities but relatively higher soil strength values (2.5–3.5 kg cm^–2) than that for the log-yard waste scrapings or the sandy soil, due to its self-cementing properties. Mixtures of soil material and ash had lower hydraulic conductivity rates (1.0 × 10^–4 cm s^–1) and lower infiltration rates compared to the sandy soil (5 × 10^–3 cm s^–1) or log-yard waste scrapings or pure ash. Water holding capacity increased with treatments containing ash. A soil–water balance model used to determine percolation rates through a 60 cm layer of treatment material showed that treatments containing a mixture of ash and soil material decreased the downward flow of water throughout the year.