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We studied the avoidance behaviour of the earthworm Eisenia fetida towards Cd, Cu, and Zn, trace elements (TEs) tested as chloride, nitrate and sulphate salts. Sub adults were exposed individually using dual-cell chambers at 20+2°C in the dark. Recordings were realised at different dates from 2h to 32h. We used filter paper and extruded water agar gel as exposure media to evaluate the contribution of the dermal and the digestive exposure routes on the avoidance reactions. Exposures to Cu or Cd (10mgmetal ionL(-1)) resulted in highly significant avoidance reactions through the exposure duration. Worms avoided Zn poorly and reactions towards Zn salts varied along the exposure. Worm sensitivity towards TEs differed between salts and this could result from differential toxicity or accessibility of these TE salts to earthworms. The anion in itself was not the determinant of the avoidance reactions since exposures to similar concentrations of these anions using calcium salts did not result in significant avoidance worm behaviour. Avoidance responses towards TEs were higher in the case of water agar exposures than in filter paper exposures. Thus, dermal contacts with TE solutions would elicit worm avoidance but signals from receptors located inside the digestive tract could reinforce this behaviour. The use of extruded water agar gels as the substrate allows checking the real sensitivity of earthworm species towards TEs since the TE concentrations leading to significant avoidance reactions were below those reported in the literature when using TE-spiked soils.

Mixed sludge from a pulp and paper mill was hydrothermally carbonized at 180–260 °C for 0.5–5 h with the use of HCl or NaOH for determining the effect of acid and base additions during sludge carbonization. Based on the results carbonization was mainly governed by dehydration, depolymerization and decarboxylation of sludge components. Additive type had a statistically significant effect on hydrochar carbon content and carbon and energy yield, of which especially energy yield increased through the use of HCl. The theoretical energy efficiencies of carbonization increased with decreasing reaction temperature, retention time and the use of HCl and suggested that the energy requirement could be covered by the energy content of attained hydrochar. The BOD5/COD-ratios of analyzed liquid samples indicated that the dissolved organic components could be treated by conventional biological methods.

International audience; To better understand the degradation of cellulose upon the formation of a tideline at the wet–dry interface when paper is suspended in water, the production of chemical species involved in oxidation reactions was studied. The quantitation of hydroperoxides and hydroxyl radicals was carried out in reverse phase chromatography using triphenylphosphine and terephthalic acid, respectively, as chemical probes. Both reactive oxygen species were found in the tideline immediately after its formation, in the range of micro-moles and nanomoles per gram of paper, respectively. The results indicate that hydroxyl radicals form for the most part in paper before the tideline experiment, whereas hydroperoxides appear to be produced primarily during tideline formation. Iron sulfate impregnation of the paper raised the production of hydroperoxides. After hygrothermal aging in sealed vials the hydroxyl radical content in paper increased significantly. When aged together in the same vial, tideline samples strongly influenced the degradation of samples from other areas of the paper (multi-sample aging). Different types of antioxidants were added to the paper before the tideline experiment to investigate their effect on the oxidation reactions taking place. In samples treated with iron sulfate or artificially aged, the addition of Irgafos 168 (tris(2,4-ditert-butylphenyl) phosphate) and Tinuvin 292 (bis(1,2,2,6,6-pentamethyl-4-piperidyl) sebacate and methyl 1,2,2,6,6-pentamethyl-4-piperidyl sebacate) reduced the concentration of hydroperoxides and hydroxyl radicals, respectively. Tinuvin 292 was also found to considerably lower the rate of cellulose chain scission reactions during hygrothermal aging of the paper.

Final effluent from a pulp and paper kraft mill was exposed to power ultrasound at 357 kHz with the aim of reducing color, turbidity, and chemical oxygen demand (COD). Absorbance measurements showed a bleaching of the effluent at wavelengths above 250 nm, indicating loss of aromatic chromophores. Effluent turbidity also decreased. Surprisingly, there was no observable decrease in COD, within experimental error. This is attributed to the presence of bicarbonate and sulfate ions in the final effluent, which react with hydroxyl radicals and effectively block the oxidation of organics in the effluent. This was demonstrated by sonicating solutions of potassium hydrogen phthalate (KHP) containing chloride, bicarbonate, or sulfate ions, which are the major inorganic ions in the final effluent studied. A solution containing only 2.3 mM KHP showed a 19% reduction in COD after 6 h of sonication. An identical solution with 200 ppm chloride also showed a 19% COD reduction. However, solutions with 700 ppm sulfate and 400 ppm bicarbonate showed COD reductions of 11% and 3%, respectively.

A novel sulfate-reducing bacterium, strain HB1(T), was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating paper-mill wastewater operated at 37 degrees C. Cells of strain HB1(T) were oval to rod-shaped, 1-1.3 microm wide and 2.6-3.5 microm long and Gram-negative. The optimum temperature for growth was 28-30 degrees C. In the presence of sulfate, the isolate was able to grow on H(2)/acetate, formate, ethanol, propionate, fumarate, succinate, butyrate, crotonate, catechol, benzoate, 4-hydroxybenzoate, palmitate and stearate. The isolate only grew on H(2) when acetate was added as a carbon source; when grown on formate, acetate was not required. Growth was also possible on pyruvate and crotonate without an electron acceptor. The isolate showed very poor growth on acetate. Thiosulfate and sulfate were used as electron acceptors. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain HB1(T) represents a novel lineage within the Deltaproteobacteria; sequence similarities between strain HB1(T) and members of other related genera were less than 91%. Strain HB1(T) was also distinguished from members of related genera based on differences in several phenotypic characteristics. It is a member of the family Desulfobacteraceae. The major cellular fatty acids of strain HB1(T) were C(16:0), iso-C(15:0), anteiso-C(15:0) and C(14:0). beta-Hydroxy fatty acids were also present in the range of C(14:0) to C(18:0), of which C(16:0) was the most abundant. The G+C content of the DNA was 55.1 mol%. Based on physiological, biochemical and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain HB1(T) is considered to represent a novel species in a new genus, for which the name Desulfatirhabdium butyrativorans gen. nov., sp. nov. is proposed. The type strain of Desulfatirhabdium butyrativorans is HB1(T) (=DSM 18734(T) =JCM 14470(T)).

Denitrification is applied in the tertiary treatment of wastewater to reduce nitrogen pollution. Fluorescence in situ hybridization (FISH), catalyzed reporter deposition (CARD)-FISH, cloning, and scanning electron microscopy (SEM) were applied to follow the evolution of the microbial composition and structure of granular sludge in chemolithotrophic denitrifying bioreactors fed with nitrate and thiosulfate. FISH oligonucleotide probes for the chemolitoautotrophic denitrifiers Thiobacillus denitrificans and Thiomicrospira denitrificans were designed and their utility tested. CARD-FISH and cloning data showed that bacterial diversity in the biofilms changed during the reactor operation. Chemoorganotrophic fermentative Gram-positive strains in the phyla, Actinobacteria and Firmicutes, were dominant in the methanogenic inoculum, both in terms of biodiversity and in number. Other significant phyla were Bacteroidetes and Chloroflexi. After 6 months of operation, Proteobacteria became dominant (83% of the clones). The diversity of Gram-positive bacteria was partially maintained although their abundance decreased notably. After 110 d of operation, the abundance of Tb. denitrificans cells increased considerably, from 1% to 35% of total DAPI-stained cells and from no isolated clones to 15% of the total clones. Tm. denitrificans only represented a minor fraction of the microorganisms in the sludge (1-4% of the DAPI-stained cells). These findings confirm that Tb. denitrificans was the dominant chemolitoautotrophic denitrifying microorganism in the bioreactors. The Archaeal diversity remained almost unchanged and it was represented mostly by Methanosaeta soehngenii. SEM results indicated a considerable loss in the integrity of the sludge granules during the operation, with risk of sludge buoyancy.

This study concerned the anaerobic treatment of five different industrial wastewaters with a diverse and complex chemical composition. The kinetics of biotransformation of this wastewater at different chemical oxygen demand (COD) were studied in a batch reactor. Wastewater from an amino acid producing industry (Fermex) and from a tank that received several types of wastewaters (collector) contained 0.83 g l(-1) and 0.085 g l(-1) sulfate, respectively. During the study period of 20 days, methane formation was observed in all types of wastewaters. Studies on COD biodegradation showed the reaction velocity was higher for Fermex wastewater and lower for collector wastewater, with values of 0.0022 h(-1) and 0.0011 h(-1), respectively. A lower methanogenic activity of 0.163 g CH4 day(-1) g(-1) volatile suspended solids (VSS) and 0.20 g CH4 day(-1) g(-1) VSS, respectively, was observed for paper producing and brewery wastewater. Adapted granular sludge showed the best biodegradation of COD during the 20-day period. The sulfate-reducing activity in pharmaceutical and collector wastewater was studied. A positive effect of sulfate-reducing activity on methanogenic activity was noted for both types of wastewaters, both of which contained sulfate ions. All reactions of methane generation for the tested industrial wastewaters were first-order. The results of this study suggest that the tested wastewaters are amenable to anaerobic treatment.

Objectives This study investigated whether previous findings of an increased risk of gliomas among workers in some pulp and paper mills could be confirmed for all Swedish pulp and paper mill workers and whether the increase could be attributed to certain occupational groups. Methods The study was based on the Swedish Cancer Environment Register, which links the incidence of cancer from 1971 to 1990 and the 1960 and 1970 census data on codes of occupation and industry for the whole population. Standardized incidence ratios (SIR) were used to estimate the risks for men in different occupations in pulp (N=28 142) and paper (N=39 169) mills in 1960, 1970 or both years as compared with those of all gainfully employed men in Sweden. Results Maintenance workers employed in pulp or paper mills in 1960, 1970 or both years, as well as pulp workers, showed an increased incidence of gliomas in 1971-1990 [SIR 1.5, 95% confidence interval (95% CI) 1.1-2.1 and SIR 1.5, 95% CI 1.0-2.2, respectively], whereas the incidence among process workers in paper mills was lower than expected (SIR 0.6, 95% CI 0.3-0.9). Taken together, all employed men in the pulp mill industry had an increased incidence of gliomas (SIR 1.3, 95% CI 1.0-1.7). Conclusions Pulp mill workers, but not paper mill workers, were found to have more gliomas in 1971-1990 than expected. There was an increased risk of gliomas among maintenance workers in both pulp and paper mills. Few risk factors for brain tumors are recognized, and the causes of the increase are not obvious.

In this paper we describe an alternative flow-chart for full treatment of wastewaters rich in organic substrates, ammonia (or organic nitrogen), and sulfate, such as those generated in fish cannery industries. Biogas generated during anaerobic pretreatment of these wastewaters is rich in hydrogen sulfide that needs to be removed to enable application of the biogas. Nitrogen elimination is traditionally achieved by subsequent nitrification and denitrification of the effluent of the anaerobic reactor. Alternatively, the hydrogen sulfide in the biogas can be applied as an electron donor in an autotrophic post-denitrification step. In order to study whether sufficient hydrogen sulfide containing biogas for denitrification was produced in the anaerobic reactor, the biogas composition as a function of the anaerobic reactor-pH was estimated based on a typical wastewater composition and chemical equilibrium equations. It is demonstrated that typical sulfate and nitrogen concentrations in fish cannery wastewater are highly appropriate for application of autotrophic post-denitrification. A literature review furthermore suggested that the kinetic parameters for autotrophic denitrification by Thiobacillus denitrificans represent no bottleneck for its application. Initial experimental studies in fixed-film reactors were conducted with sodium sulfide and nitrate as an electron donor-acceptor couple. The results revealed that only moderate volumetric treatment capacities (1 g-NO3- N l(-1) day(-1)) could be achieved. Mass balances suggested that incomplete sulfide oxidation to elemental sulfur occurred, limiting biomass retention and the treatment capacity of the reactor. Future research should clarify the questions concerning product formation from sulfide oxidation.

Stringent environmental laws in Europe and Northern America lead to the development towards closure of the process water streams in pulp and paper mills. Application of a "zero-discharge" process is already a feasible option for the board and packaging paper industry, provided in-line treatment is applied. Concomitant energy conservation inside the mill results in process water temperatures of 50-60 degrees C. Thermophilic anaerobic treatment complemented with appropriate post-treatment is considered as the most cost-effective solution to meet re-use criteria of the process water and to keep its temperature. In the proposed closed-cycle, the anaerobic treatment step removes the largest fraction of the biodegradable COD and eliminates "S" as H2S from the process stream, without the use of additional chemicals. The anaerobic step is regarded as the only possible location to bleed "S" from the process water cycle. In laboratory experiments, the effect of upward liquid velocity (Vupw) and the specific gas loading rate (Vgas) on the S removal capacity of thermophilic anaerobic bio-reactors was investigated. Acidifying, sulphate reducing sludge bed reactors were fed with partly acidified synthetic paper mill wastewater and were operated at 55 degrees C and pH 6. The reactors were operated at organic loading rates up to 50 g COD.l-1.day-1 at COD/SO4(2-) ratios of 10. The effect of Vupw was researched by comparing the performance of a UASB reactor operated at 1.0 m.h-1 and an EGSB reactor, operated at 6.8 m.h-1. The Vupw had a strong effect on the fermentation patterns. In the UASB reactor, acidification yielded H2, acetate and propionate, leading to an accumulation of reducing equivalents. These were partly disposed of by the production of n-butyrate and n-valerate from propionate. In the EGSB reactor net acetate consumption was observed as well as high volumetric gas (CO2 and CH4) production rates. The higher gas production rates in the EGSB reactor resulted in higher S-stripping efficiencies. The effect of Vgas was further researched by comparing 2 UASB reactors which were sparged with N2 gas at a specific gas loading rate of 30 m3.m-2.day-1. In contrast to the regular UASB reactors, the gas-supplied UASB showed a more stable performance when the organic loading rates were increased. Also, the H2S stripping efficiency was 3-4 times higher in the gas-supplied UASB, reaching values of 67%. Higher values were not obtained owing to the relatively poor sulphate reduction efficiencies.

Measurement of oxalate in urine is important for the diagnosis of primary hyperoxaluria (McKusick 259900) and the secondary forms produced by excessive intake or abnormal intestinal absorption of oxalate (1). Determination of glycolic acid is essential for the diagnosis of primary hyperoxaluria type 1. Finally, to estimate the risk of stone formation in calcium oxalate urolithiasis and nephrocalcinosis, simultaneous determination not only of calcium but also of citrate (a potent inhibitor of calcium oxalate and calcium phosphate crystallization) and other constituents (electrolytes, phosphate and sulfate) is required to calculate urinary calcium saturation (2). Ion-chromatography HPLC (3)(4) and specific enzymatic assays (5)(6) are available only in specialized laboratories. In addition, preservation and storage of liquid samples may influence the stability of oxalate and glycolate (7). Use of urinary filter spots is a practical alternative for the collection and safe transport of samples to be analyzed for many metabolic disorders. To evaluate the age-related changes of oxalate, glycolate, citrate, and sulfate in a pediatric population, we developed an automated ion-chromatography system for the simultaneous measurement of these anions in urine and established their reference values for liquid urine samples as well as for dried urine on filter paper. We studied 20 individuals from each age group ( 16 years), all without renal or metabolic disease, all on a routine food intake at the time of study. The urines were preserved with 6 mol/L HCl at pH 1–2 to prevent nonenzymatic conversion of ascorbate to oxalate. Filter paper strips (3 × 5 …

Toxic effects of unbleached (sulfate or sulfite) and bleached (sulfate) paper mill effluents were studied in a primary culture of rainbow trout liver cells. The effluents and control water from a clean area were extracted with diethyl ether and added to the cultures dissolved in dimethyl sulfoxide. Plasma membrane integrity was studied by measuring lactate dehydrogenase (LDH) leakage. The cellular content of glutathione (GSH) was used as an indicator of oxidative stress and the formation of reactive intermediates. Dose-response studies indicated that unbleached effluents contained more potent toxic substances than bleached effluents. Both unbleached and bleached effluents contained organic diethyl ether-extractable substances which increased cytochrome P450-dependent 7-ethoxyresorufin-O-deethylase (EROD) activities. The inducing effects were seen at concentrations substantially lower than those decreasing GSH content and increasing LDH leakage. Possible EROD inducing substances in bleached effluents are chlorinated organic compounds. Inducing compounds in unbleached effluents are yet to be identified. Furthermore, at higher concentrations the effluents contained substances that inhibited the cytochrome P450 system. The results show that the trout primary hepatocyte cultures afford a convenient in vitro method for screening cytochrome P450 inducing components extracted from industrial effluents to investigate mechanisms by which wastewaters cause injury in cells.

Biologically active 14 C-labeled purified protein derivative ( 14 C-PPD) has been prepared from the culture filtrates of seven species of mycobacteria, namely Mycobacterium tuberculosis Johnston strain (PPD), M. bovis BCG (PPD-BCG), M. avium (PPD-A), M. kansasii (PPD-Y), M. intracellulare (PPD-B), M. scrofulaceum (PPD-G), and M. fortuitum (PPD-F). These mycobacteria were grown in a culture medium containing a mixture of 14 C-labeled amino acids. The yield and specific radioactivity of the PPD, of the nucleic acid, of the bacterial cells, and of the CO 2 developed during growth have been determined for each of the seven species of mycobacteria. Although the yields of 14 C-PPD antigens differed greatly for the different species of mycobacteria tested, their specific radioactivities were similar. The 14 C-PPD antigens have been used as a means to measure their adsorption to glass. When glass ampoules containing dilute solutions (0.001 mg of PPD per ml) of these PPD antigens (PPD, PPD-BCG, PPD-A, PPD-Y, PPD-G, PPD-B, and PPD-F) were stored for 12 months at 5 C, it was found that they all adsorbed equally well to glass surfaces. In fact, regardless of the origin of the PPD, a loss due to adsorption of about 90% occurred during the first month of storage, and thereafter the PPD content remained practically constant for the rest of the duration of the storage period. The addition of 0.0005% Tween 80 to the PPD solutions effectively reduced the adsorption to glass of most PPD antigens. However, adsorption of PPD-BCG was not quite so effectively prevented, even when the Tween 80 concentration was increased from 0.0005 to 0.0005%.

1. After the administration of large doses of androsterone, epiandrosterone, dehydroepiandrosterone and testosterone to mice, females excreted more of the dose conjugated with sulphuric acid than did males. 2. Liver slices from female mice conjugated androgens with sulphuric acid to a greater extent than did slices from males. 3. Sulphotransferase preparations from livers of female rats and mice catalysed the formation of dehydroepiandrosterone sulphate at a faster rate than preparations from livers of the male animals. 4. A possible explanation for the observed sex differences is discussed.

Cell-free extract (crude extract) of Agrobacterium tumefaciens grown on d -glucuronate or d -glucarate converts d -glucarate and galactarate to a mixture of 2-keto-3-deoxy- and 4-deoxy-5-keto- d -glucarate. These compounds are then converted by partially purified crude extract to an intermediate tentatively identified as 2,5-diketoadipate. The same enzyme preparation further decarboxylates this intermediate to α-ketoglutarate semialdehyde, which is subsequently oxidized in a nicotinamide adenine dinucleotide-dependent reaction to α-ketoglutaric acid. Since A. tumefaciens converts d -glucuronic acid to d -glucarate, a pathway from d -glucuronate to α-ketoglutarate in A. tumefaciens was determined.