Trace metal elements vaporization and phosphorus recovery during sewage sludge thermochemical treatment - A review.

Phosphorus (P) plays essential roles in crops growth. Natural mineral sources of phosphate are non-renewable, overexploited and unevenly distributed worldwide, making P a strategic resource for agricultural systems. The search for sustainable ways to secure P supply for fertilizer production has therefore become a critical issue worldwide. Sewage sludge (SS) is an organic waste material considered as a key alternative source of P. Switzerland and the European Union are about to make it mandatory to recover P from SS or its treatment residues. Among the many technical options to achieve this objective, SS thermochemical treatments spiked with Cl-donors appear as a promising approach to recover P from SS and separate it from mineral pollutants such as trace metal elements (TME). The purpose of Cl-donor additives is to fix P within the mineral residues, possibly in bioavailable P species forms, while promoting TME vaporization by chlorination mechanisms. This review paper compares the various thermochemical treatments investigated worldwide over the past two decades. The influence of process conditions and Cl-donor nature is discussed. The presented results show that, except for nickel and chromium, most TME can be significantly vaporized during a high temperature treatment (over 900 °C) with Cl addition. In addition, the fixation rate and solubility of P is increased when a Cl-donor such as MgCl2 is added.

Comparative characterization of surface sludge deposits from fourteen French Vertical Flow Constructed Wetlands sewage treatment plants using biological, chemical and thermal indices.

Due to their design and mode of operation, French Vertical Flow Constructed Wetlands (VFCWs) accumulate suspended solids from the inflow wastewater in the form of a sludge layer at the surface of the first filter. In order to maintain the treatment performance over the long term, the characteristics of the sludge deposits and their evolution have to be well described. In this objective, a panel of sludge deposit samples taken from 14 French VFCW sewage treatment plants was investigated. Elemental composition and organic matter content, nature and reactivity were analyzed. Results clearly revealed two categories of sludge deposits, namely the "young-age plants" type (1 year of operation and less) and the "mature plants" type (3 years of operation and more). Sludge deposits from the "mature plants" exhibited same biological, physical and chemical properties. Their organic matter was globally less abundant, more humified and less biodegradable than in the young-age plants type. Their overall contents in trace metals were also higher, although in a limited manner. The effect of additional treatments, particularly FeCl3 injection for phosphorus precipitation, was observable in the "young-age plants" group. Finally, the sludge deposits sampled from one particular plant with specific operating conditions were found to exhibit very different characteristics from those of either groups identified. This observation underlined the influence of local conditions on the typology of the sludge deposits.

Genome scans on experimentally evolved populations reveal candidate regions for adaptation to plant resistance in the potato cyst nematode Globodera pallida.

Improving resistance durability involves to be able to predict the adaptation speed of pathogen populations. Identifying the genetic bases of pathogen adaptation to plant resistances is a useful step to better understand and anticipate this phenomenon. Globodera pallida is a major pest of potato crop for which a resistance QTL, GpaVvrn , has been identified in Solanum vernei. However, its durability is threatened as G. pallida populations are able to adapt to the resistance in few generations. The aim of this study was to investigate the genomic regions involved in the resistance breakdown by coupling experimental evolution and high-density genome scan. We performed a whole-genome resequencing of pools of individuals (Pool-Seq) belonging to G. pallida lineages derived from two independent populations having experimentally evolved on susceptible and resistant potato cultivars. About 1.6 million SNPs were used to perform the genome scan using a recent model testing for adaptive differentiation and association to population-specific covariables. We identified 275 outliers and 31 of them, which also showed a significant reduction in diversity in adapted lineages, were investigated for their genic environment. Some candidate genomic regions contained genes putatively encoding effectors and were enriched in SPRYSECs, known in cyst nematodes to be involved in pathogenicity and in (a)virulence. Validated candidate SNPs will provide a useful molecular tool to follow frequencies of virulence alleles in natural G. pallida populations and define efficient strategies of use of potato resistances maximizing their durability.

Pilot-scale study of vertical flow constructed wetland combined with trickling filter and ferric chloride coagulation: influence of irregular operational conditions.

The aim of this study was to characterize the efficiency of an intensified process of vertical flow constructed wetland having the following particularities: (i) biological pretreatment by trickling filter, (ii) FeCl3 injection for dissolved phosphorus removal and (iii) succession of different levels of redox conditions along the process line. A pilot-scale set-up designed to simulate a real-scale plant was constructed and operated using real wastewater. The influences of FeCl3 injection and water saturation level within the vertical flow constructed wetland stage on treatment performances were studied. Three different water saturation levels were compared by monitoring: suspended solids (SS), total phosphorus (TP), dissolved chemical oxygen demand (COD), ammonium, nitrate, phosphate, iron, and manganese. The results confirmed the good overall efficiency of the process and the contribution of the trickling filter pretreatment to COD removal and nitrification. The effects of water saturation level and FeCl3 injection on phosphorus removal were evaluated by analysis of the correlations between the variables. Under unsaturated conditions, good nitrification and no denitrification were observed. Under partly saturated conditions, both nitrification and denitrification were obtained, along with a good retention of SSs. Finally, under saturated conditions, the performance was decreased for almost all parameters.

Isotopic investigation of the colloidal mobility of depleted uranium in a podzolic soil.

The mobility and colloidal migration of uranium were investigated in a soil where limited amounts of anthropogenic uranium (depleted in the 235U isotope) were deposited, adding to the naturally occurring uranium. The colloidal fraction was assumed to correspond to the operational fraction between 10 kDa and 1.2 µm after (ultra)filtration. Experimental leaching tests indicate that approximately 8-15% of uranium is desorbed from the soil. Significant enrichment of the leachate in the depleted uranium (DU) content indicates that uranium from recent anthropogenic DU deposit is weakly bound to soil aggregates and more mobile than geologically occurring natural uranium (NU). Moreover, 80% of uranium in leachates was located in the colloidal fractions. Nevertheless, the percentage of DU in the colloidal and dissolved fractions suggests that NU is mainly associated with the non-mobile coarser fractions of the soil. A field investigation revealed that the calculated percentages of DU in soil and groundwater samples result in the enhanced mobility of uranium downstream from the deposit area. Colloidal uranium represents between 10% and 32% of uranium in surface water and between 68% and 90% of uranium in groundwater where physicochemical parameters are similar to those of the leachates. Finally, as observed in batch leaching tests, the colloidal fractions of groundwater contain slightly less DU than the dissolved fraction, indicating that DU is primarily associated with macromolecules in dissolved fraction.

Physical-chemical characterization of sludge and granular materials from a vertical flow constructed wetland for municipal wastewater treatment.

The use of vertical flow constructed wetlands (VFCWs) is well developed in France and other countries for the treatment of wastewaters from small communities. The patented Azoé® process has been developed by a French company, SCIRPE, in order to improve denitrification and phosphorus removal as compared to classical VFCWs. It includes a biological trickling filter pretreatment followed by two stages of partially flooded VFCW. The performances of partially flooded VFCW are well demonstrated for the removal of organic matter and nitrogen. The system is now being considered for phosphorus removal as well. In this article, sludge and granular materials sampled from the filters of a municipal plant where the Azoé® system has been operated for 8 years were analyzed in order to provide data that may contribute to a better understanding of the dynamics of phosphorus retention. Elemental analyses showed that phosphorus was predominantly captured in the sludge layer accumulated at the surface of the first stage. The progressive mineralization of the sludge over time was also clearly highlighted. The phosphate phases were mainly associated with iron and calcium. The transport of phosphorus via the migration of fine particles through the porous medium in the first stage was also observed.

The case for re-evaluating the upper limit value for selenium in drinking water in Europe.

Selenium is an essential trace element for life, which can be toxic for humans when intakes reach a certain amount. Therefore, since the margin between healthy intake and toxic intake is narrow, the selenium concentration of tap water is a parameter that must be monitored because of its potential for increased intake. The present work gives an overview of the different approaches used to calculate safe limits for selenium. As recommended by WHO, the guidelines for drinking water form the basis of national legislated standards for drinking water. Before setting a maximum acceptable level in drinking water, it is necessary to take into account the total intake of selenium in both food and beverage. The limit value of 10 microg l(-1) for drinking water laid down in the European regulations for all countries should be adapted depending on geographic area, as previously recommended by WHO.

High-performance liquid chromatographic-inductively coupled plasma mass spectrometric evidence for Se-"alliins" in garlic and onion grown in Se-rich soil.

Garlic and onion, are well known for their medical value, especially in against cancer and anticardiovacular diseases. "Alliins" (S-alk(en)yl-L-cysteine sulphoxides) are sources of major active compounds in Allium plants. Se incorporation into garlic significantly increases activities of garlic in cancer prevention and inhibition. Selenomethionine, selenocysteine and Se-methylselenocysteine have been identified in garlic and onion. Previously we identified gamma-glutamyl-Se-methyl-L-selenocysteine, in extracts of garlic cultivated in Se-rich soil [Med. Res. Rev. 16 (1) (1996) 111], suggesting the possible existence of Se-alk(en)yl-L-cysteine selenoxides (Se-"alliins") in garlic. Several comparative experiments were carried out to demonstrate the existence of Se-"alliins" in Se-enriched garlic and onion. We found that there was one similar time-dependent Se signal in HPLC-inductively coupled plasma MS chromatograms of cold-water extracts of freeze-dried garlic powder and fresh garlic. This signal was lost when the extracts of garlic powder and fresh garlic were stored for 1 day at >4 degrees C, but remained in fresh onion extract at the same storage conditions. These phenomena and possible mechanisms are discussed. An additional experiment showed that Allium species cultivated in Se-rich soil might contain two different Se-"alliins".

Determination of phytochelatins by capillary zone electrophoresis with electrospray tandem mass spectrometry detection (CZE-ES MS/MS).

The coupling of capillary zone electrophoresis with electrospray mass spectrometry was optimized for the direct determination of phytochelatins (PCs) in extracts obtained from cells and plants that had been exposed to metal stress. Gluthathione and phytochelatins belonging to the different families (gamma Glu-Cys)nGly (n-PC), (gamma Glu-Cys)nSer, (gamma Glu-Cys)n beta Ala and (gamma Glu-Cys)n were separated in an uncoated capillary at pH 4 using a 5 mM ammonium acetate buffer, and detected by electrospray (ES) MS in the full scan mode (300-1100 u). The use of on-line tandem MS detection in the product ion scan mode of putative protonated molecules of PCs allowed the unambiguous confirmation of the identity of the compounds detected by ES MS. The operational conditions were optimized and the figures of merit were evaluated using n-PC2, n-PC3 and n-PC4 standards purified from a mixture obtained after the reaction of glutathione in the presence of Cd2+ and the enzyme PC-synthase. The method was applied to the characterization of bioinduced ligands in cell cultures of soybeans (Glycine max) and in rice (Oryza sativa) roots without the need for a preliminary sample cleanup by size-exclusion and/or reversed phase chromatography.

Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion.

BACKGROUND: Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.

The copines, a novel class of C2 domain-containing, calcium-dependent, phospholipid-binding proteins conserved from Paramecium to humans.

In an attempt to identify proteins that might underlie membrane trafficking processes in ciliates, calcium-dependent, phospholipid-binding proteins were isolated from extracts of Paramecium tetraurelia. The major protein obtained, named copine, had a mass of 55 kDa, bound phosphatidylserine but not phosphatidylcholine at micromolar levels of calcium but not magnesium, and promoted lipid vesicle aggregation. The sequence of a 920-base pair partial cDNA revealed that copine is a novel protein that contains a C2 domain likely to be responsible for its membrane active properties. Paramecium was found to have two closely related copine genes, CPN1 and CPN2. Current sequence data bases indicate the presence of multiple copine homologs in green plants, nematodes, and humans. The full-length sequences reveal that copines consist of two C2 domains at the N terminus followed by a domain similar to the A domain that mediates interactions between integrins and extracellular ligands. A human homolog, copine I, was expressed in bacteria as a fusion protein with glutathione S-transferase. This recombinant protein exhibited calcium-dependent phospholipid binding properties similar to those of Paramecium copine. An antiserum raised against a fragment of human copine I was used to identify chromobindin 17, a secretory vesicle-binding protein, as a copine. This association with secretory vesicles, as well the general ability of copines to bind phospholipid bilayers in a calcium-dependent manner, suggests that these proteins may function in membrane trafficking.

Expression of a cDNA encoding the wheat CM16 protein in Escherichia coli.

The wheat kernel CM16 protein, a subunit of the heterotetrameric insect alpha-amylase inhibitor that has been involved in the technological quality of wheat-products, was produced in Escherichia coli. Cloning of the cDNA part encoding the mature protein in a pET expression plasmid, under the control of a promoter for the bacteriophage T7 RNA polymerase, allows the synthesis of large amounts of the CM16 protein in the bacteria. Upon induction with isopropyl thiogalactopyranoside the recombinant protein accumulates in insoluble inclusion bodies. Solubilization with 6 M urea containing 0.5 mM dithiothreitol, followed by slow elimination of the denaturing agents by step dialysis, results in a significant recovery of the recombinant protein in a soluble, monomeric form. Characterization of the protein was done by automated Edman degradation and total amino acid determination. The recombinant protein in comparison with the one isolated from wheat exhibits a Met extension at the N-terminus that was introduced in the construction for translation initiation. The CM16 protein produced in this manner has the advantage over wheat purified protein of not being contaminated with other proteins from the same family and constitutes adequate material for further analysis of the technological properties of the protein in wheat-derived products.

Fructose-induced enhanced mitogenicity of diploid human cells: possible relationship with cell differentiation.

Fructose strongly stimulates the growth of normal diploid human skin fibroblasts (SFs) and induces marked changes in their morphology and lipid accumulation. This mitogenic effect occurs despite very low fructose consumption and depends on the presence of glutamine. The cell kinetics of cultured fructose-fed human skin fibroblasts were different from those fed on glucose: in the presence of fructose a high proliferative index persisted at Day 14 of culture and the duration of the total cell cycle and of the G1 + 1/2 M and S phases was slightly shorter. The mitogenic effect of fructose on SF was largest in the presence of human serum: it was small or undetectable when fibroblasts were cultured in media supplemented with dialyzed human serum, fetal bovine serum, or serum substitutes. This suggests that serum growth factor(s) mediate the mitogenic effect of fructose. Only normal diploid human cells seem to be sensitive to this mitogenic effect of fructose: the long-term growth of normal human liver cells on fructose was slightly better or similar to that on glucose. In contrast, fructose could only support limited growth of hamster fibroblastic Nil cells and of a transformed human fibroblastic line, which grew better with glucose.

Effects of polyunsaturated fatty acids on human and rat cells. In vitro versus in vivo experiments.

Human infant skin fibroblasts and liver cells were subcultured with 250 microM PUFA (polyunsaturated fatty acid), and primary cultures of glial brain cells from new-born rats with 100 microM; oleic acid was added to controls. Minimum essential medium (MEM) supplemented with bovine serum was used as a reference. During the short-term experiment (18-24 h), control liver cells showed a regular increase in protein level, while protein increment was more rapid in linoleic and especially in arachidonic acid-treated cells, but only for the first 3 hours. During the long-term experiment (7 d), control skin fibroblasts showed a faster growth rate (increase in number of cells) than reference or fibroblasts cultured with the added PUFAs. Lipid droplets were seen in the PUFA-treated liver cells and skin fibroblasts, and ultrastructural modifications were observed in fibroblasts, but without growth rate alteration. During the long-term experiment (2 w), control glial brain cells showed faster protein increment (measuring growth rate) than PUFA-treated cells, particularly than arachidonic acid-treated cells. HMGR (3-hydroxy-3-methylglutaryl-CoA reductase) activity, determined after 6 h (liver cells) or 1 and 2 w (brain cells) of culture, was low in controls and reference, whilst higher in PUFA-treated-cells, and was especially high in arachidonic acid-treated brain cells. The present study indicates than the high HMGR activity may correspond to cultures of cells rapidly stopped in their protein increment, and to cultures of cells showing a slow rate of proliferation. This contrasts with results obtained from in vivo experiments; it also emphasizes the high mevalonate (MVA) level as a possible sign of nutritional medium imbalance.(ABSTRACT TRUNCATED AT 250 WORDS)

Glutamine dependency of human skin fibroblasts: modulation by hexoses.

The combined effects of carbohydrates and glutamine were investigated in diploid strains of normal human skin fibroblasts cultured for 21 days under eight different culture conditions: hexose-free medium or medium containing D-glucose, D-galactose, or D-fructose, with or without added glutamine. Cell growth, hexose consumption, lactate production, intracellular glycogen content and extracellular amino acid levels were measured every third to fourth day. In the presence of glutamine, cells reached a higher saturation density in fructose medium than in glucose or galactose medium but per cell consumption of fructose and galactose was much less than that of glucose. Consumption of all three carbohydrates per unit cell growth exhibited three distinct phases: Days 1-3, 3-10, and 10-20, respectively. In the absence of glutamine the rate of cell growth was not altered in glucose or galactose medium, but slowed down considerably in fructose medium. Glutamine deprivation also led to changes in hexose consumption. In hexose-free media the cell growth rate at first was very slow, but rose after 2 or 3 weeks of culture. The levels of extracellular nonessential amino acids varied according to medium and growth phase. One of the most exciting findings was that human fibroblasts are able to maintain a slight excess of glutamine in all media not supplemented with glutamine and, more surprisingly, to synthesize it in a medium containing galactose and glutamine.

Bioassay alpha spectrometry: energy resolution as a function of sample source preparation and counting geometry.

Alpha particle counting is based on the response of an electronic counting system to an incident alpha particle. Alpha spectrometry is used in our employee surveillance bioassay program to measure the concentration of isotopes of Am, Pu and U contained on sample source preparations. Nuclides of Am, Pu and U are separated from the sample matrix by anion exchange and are electroplated on a stainless steel disc (Gautier and Gladney 1986). The plated source diameter is 12.7 mm. A tracer is added to the sample before anion exchange as a quality control procedure to provide a measure of chemical yield. Tracer alpha-particle emissions are recorded in a preassigned calibrated area of the energy spectrum and chemical recovery is calculated by the ratio of tracer counts per second divided by the tracer activity in becquerels (Bq). Percent tracer recovery may also be calculated by introducing the average counting efficiency factor in the denominator. Tracer yield is then used to provide a reliable estimate of the sample's analyte counts that are recorded in other preassigned energy dependent areas of the spectrum. The tracer spectrum in the presence or absence of other nuclides also provides evidence of the performance characteristics of the alpha spectrometer, for example, chamber vacuum and electronics. Electroplated samples are counted in any one of 96 detectors. The backgrounds of these detectors are maintained at less than 3 counts per 70,000 s over a 190 keV energy window to provide a limit of detection of less than 0.37 mBq per sample at the 95% confidence level. In this paper, resolution of the photopeak is shown to be a function of the source to detector distance and a function of degraded alpha energies due to Fe or other extraneous materials on the plated surface. Since our program is concerned only with low activity samples (less than 0.2 Bq) we encounter no significant problem with tailing when the resolution is less than 50 keV FWHM.

Syndromic paucity of interlobular bile ducts (Alagille syndrome or arteriohepatic dysplasia): review of 80 cases.

We have observed two types of paucity of interlobular bile ducts (PILBD) in children with chronic cholestasis: the syndromic type, which is more frequent (80 cases), and the nonsyndromic type (31 cases). Study of patients with syndromic PILBD has enabled us to recognize five major features: peculiar facies (95%), chronic cholestasis (91%), posterior embryotoxon (88%), butterfly-like vertebral arch defects (87%), and peripheral pulmonary artery hypoplasia or stenosis, either isolated or associated with complex cardiovascular abnormalities (85%). By observing these major features, it is possible to differentiate the "complete" syndrome, in which all five features are present (26 cases), from the "partial" syndrome, in which only four (42 cases) or three (12 cases) major features are present. Other less frequent features were observed in patients with complete or partial syndrome: growth retardation (50%), mental retardation (16%), renal disturbances, other vascular malformations, bone abnormalities, high-pitched voice, and delayed puberty. Death occurred in 21 (26%) patients with syndromic PILBD. Therapy consisted of supplementation of medium-chain triglycerides and fat-soluble vitamins and administration of cholestyramine or phenobarbital. An autosomal dominant mode of transmission, with variable penetrance, seems likely.

In-vitro responses to ascorbate and manganese in fibroblasts from a patient with prolidase deficiency and iminodipeptiduria: cell growth, prolidase activity and collagen metabolism.

After successful ascorbate and manganese treatment of a female patient with prolidase deficiency and iminodipeptiduria, we attempted to explain the mechanism of action of these drugs in vitro, using them preferentially on skin fibroblasts. Since in vivo, ascorbate and manganese seemed to be responsible for both biochemical and clinical improvement, they were also expected to activate prolidase activity in vitro. Cell growth and prolidase activity were accordingly observed in fibroblast cultures supplemented with these compounds. It seemed that only ascorbate accounted for the successful in vivo response. To understand the mechanism involved, we studied collagen metabolism and found a decreased proline pool, a massive increase of rapidly degraded collagen and moderate enhancement of type III collagen and type I trimer in the patient's fibroblasts. We believe that ascorbate allowed the prolidase-deficient cells to maintain a normal collagen pool by increasing collagen synthesis. Both the massive increase in cell growth in response to ascorbate and the bad response as regards the quality of the collagen produced confirm the secondary nature of this mechanism. However, the relationship between accelerated collagen catabolism and prolidase deficiency remains unclear.