Brewery wastewater treatment using MBR coupled with nanofiltration or electrodialysis: biomass acclimation and treatment efficiency.

Breweries release significant amounts of wastewater loaded with various organic and mineral materials. Prior studies of membrane bioreactor (MBR) wastewater treatment have been conducted with very little interest granted to the conditions of biomass acclimation. This study displays biomass behavior during brewery wastewater treatment by an aerobic MBR. In addition, nanofiltration and electrodialysis have been studied as potential post-treatment to decrease mineral concentrations and permit further water reuse for agriculture. An anoxic/aerobic laboratory MBR, associated with a flat sulfonated polyether membrane was used for synthetic brewery wastewater treatment. Biomass acclimation was performed using a feeding substrate. Organic concentrations in the MBR influent varied from 700 mg COD/L to 10,600 mg COD/L (COD: chemical oxygen demand) for 110 days. The results indicate a good acclimation to effluent with high salts and organic matter loads. Steady evolution of biomass concentration and activities was achieved after 90 days of operation. A reduction of COD of around 95% was obtained with MBR and up to 99% with nanofiltration post-treatment for the reconstructed brewery effluent with an organic loading rate of 7 g COD/L·d and a solid and hydraulic retention time of 30 days and 36 hours. A good reduction of the salt content was also recorded primarily with the nanofiltration and electrodialysis processes.

Cost minimization in a full-scale conventional wastewater treatment plant: associated costs of biological energy consumption versus sludge production.

Energy consumption and sludge production minimization represent rising challenges for wastewater treatment plants (WWTPs). The goal of this study is to investigate how energy is consumed throughout the whole plant and how operating conditions affect this energy demand. A WWTP based on the activated sludge process was selected as a case study. Simulations were performed using a pre-compiled model implemented in GPS-X simulation software. Model validation was carried out by comparing experimental and modeling data of the dynamic behavior of the mixed liquor suspended solids (MLSS) concentration and nitrogen compounds concentration, energy consumption for aeration, mixing and sludge treatment and annual sludge production over a three year exercise. In this plant, the energy required for bioreactor aeration was calculated at approximately 44% of the total energy demand. A cost optimization strategy was applied by varying the MLSS concentrations (from 1 to 8 gTSS/L) while recording energy consumption, sludge production and effluent quality. An increase of MLSS led to an increase of the oxygen requirement for biomass aeration, but it also reduced total sludge production. Results permit identification of a key MLSS concentration allowing identification of the best compromise between levels of treatment required, biological energy demand and sludge production while minimizing the overall costs.

New urban wastewater treatment with autotrophic membrane bioreactor at low chemical oxygen demand/N substrate ratio.

The potential for total nitrogen removal from municipal wastewater has been evaluated in an autotrophic membrane bioreactor running with a low chemical oxygen demand (COD)/N ratio to simulate its combination with an upstream physicochemical process that retains a large proportion of organic matter. The tests were conducted in a laboratory scale submerged membrane bioreactor loaded with a synthetic influent. Nitrogen loading rate was 0.16 kgN-NH4+.m(-3).d(-1) and sodium acetate was added as a carbon source. Results have shown that nitrogen elimination can reach 85% for a COD/N ratio of 5, with COD removal exceeding 97%. However, a COD/N ratio of 3.5 was found to be the limiting factor for successfully reaching the overall target value of 10 mgN.L(-1) in the effluent. Nevertheless, low COD/N ratios make it possible to work with low total suspended solid concentrations in the bioreactor, which greatly facilitates membrane fouling control by a simple aeration and backwashing strategy.

Short-term effect of reclaimed wastewater quality gradient on soil microbiome during irrigation.

To investigate the effect of wastewater (WW) treatment on soil bacterial communities, water of different quality was used to irrigate eight lettuces per tank: raw municipal wastewater (RWW), WW treated with an aerated constructed wetland (CWW) and WW treated with a membrane bioreactor (MBW), and tap water (TW). The physicochemical and microbiological characteristics (quality indicators) of these water types were characterized, and the water and soil bacterial communities were monitored by quantitative PCR (qPCR) and 16S rRNA gene sequencing. Despite marked differences in microbial load and diversity of waters, soil communities remained remarkably stable after irrigation. Microbial biomass was increased only in soils irrigated with RWW. At the end of the irrigation period (day 84), soil and water shared a large fraction of their bacterial communities, from 43 % to 70 %, depending on the water quality, indicating a transfer of bacterial communities from water to soil. Overall, the relative abundance of Proteobacteria and Acidobacteria was increased and that of Actinobacteria was decreased in soils irrigated with MBW, CWW and even more with RWW. Multivariate ordination clearly separated soils in three groups: soils irrigated with the cleanest water (TW), with treated WW (MBW and CWW), and with untreated WW (RWW). Nitrifying, denitrifying, and nitrogen-fixing bacteria were quantified by qPCR targeting amoA, narG, and nifH, respectively. Nitrifying bacteria were the most affected by the water quality, as indicated by amoA copy number increase in RWW-irrigated soil and decrease in CWW-irrigated soil. Overall, the abundance of all three genes was positively influenced by RWW treatment. In conclusion, the 84 days of irrigation influenced the soil microbial communities, and the impact depended on the quality of the used water.

Expression of GCIP in transgenic mice decreases susceptibility to chemical hepatocarcinogenesis.

Transcription factors with helix-loop-helix (HLH) motif play critical roles in controlling the expression of genes involved in lineage commitment, cell fate determination, proliferation, and tumorigenesis. To examine whether the newly identified HLH protein GCIP/CCNDBP1 modulates cell fate determination and plays a role in hepatocyte growth, proliferation, and hepatocarcinogenesis, we generated transgenic mice with human GCIP gene driven by a liver-specific albumin promoter. We demonstrated that in GCIP transgenic mice, the overall liver growth and regeneration occurred normally after liver injury induced by carbon tetrachloride (CCl4). In the diethylnitrosamine (DEN)-induced mouse hepatocarcinogenesis, we demonstrated that overexpression of GCIP in mouse liver suppressed DEN-induced hepatocarcinogenesis at an early stage of tumor development. The number of hepatic adenomas at 24 weeks was significantly lower or not detected in GCIP transgenic male mice compared to the control mice under the same treatment. Although GCIP has little inhibition on the number of hepatic tumors at later stages (40 weeks), hepatocellular tumors in GCIP transgenic mice are smaller and well-differentiated compared to the poorly differentiated tumors in wild-type mice. Furthermore, we demonstrate that GCIP functions as a transcriptional suppressor, regulates the expression of cyclin D1, and inhibits anchorage-independent cell growth and colony formation in HepG2 cells, suggesting a significant role of GCIP in tumor initiation and development.

Biliary excretion of iron from hepatocyte lysosomes in the rat. A major excretory pathway in experimental iron overload.

In these experiments, we assessed the role of hepatocyte lysosomes in biliary excretion of iron. We loaded rats with iron by feeding 2% carbonyl iron and collected bile for 24 h via bile fistulae from iron-loaded and control rats. In additional rats, bile was collected before and after the administration of colchicine. Rats were then killed and their livers were homogenized and fractionated for biochemical analyses or processed for electron microscopy and x-ray microanalysis. Inclusion of 2% carbonyl iron in the diet caused a 45-fold increase (P less than 0.001) in hepatic iron concentration compared with controls (1,826 +/- 159 vs. 38 +/- 6.7 micrograms/g liver, mean +/- SE). Electron microscopy with quantitative morphometry and x-ray microanalysis showed that the excess iron was sequestered in an increased number of lysosomes concentrated in the pericanalicular region of the hepatocyte. Iron loading was also associated with a twofold increase in biliary iron excretion (4.06 +/- 0.3 vs. 1.75 +/- 0.1 micrograms/g liver/24 h; P less than 0.001). In contrast, the biliary outputs of three lysosomal enzymes were significantly lower (P less than 0.0005) in iron-loaded rats compared with controls (mean +/- SE) expressed as mU/24 h/g liver: N-acetyl-beta-glucosaminidase, 26.7 +/- 4.6 vs. 66.2 +/- 13.4; beta-glucuronidase, 10.1 +/- 1.3 vs. 53.2 +/- 17.9; beta-galactosidase, 8.9 +/- 1.0 vs. 15.4 +/- 2.3. In iron-loaded rats but not in controls, biliary iron excretion was coupled to the release into bile of each of the three lysosomal hydrolases as assessed by linear regression analysis (P less than 0.001). In contrast, no relationships were found between biliary iron excretion and the biliary outputs of a plasma membrane marker enzyme (alkaline phosphodiesterase I) or total protein. After administration of colchicine, there was a parallel increase in biliary excretion of iron and lysosomal enzymes in iron-loaded rats, but not controls. We interpret these data to indicate that, in the rat, biliary iron excretion from hepatocyte lysosomes is an important excretory route for excess hepatic iron.

The Kex2p proregion is essential for the biosynthesis of an active enzyme and requires a C-terminal basic residue for its function.

The Saccharomyces cerevisiae prohormone-processing enzyme Kex2p is biosynthesized as an inactive precursor extended by its N-terminal proregion. Here we show that deletion of the proregion renders Kex2p inactive both in vivo and in vitro. Absence of the proregion impaired glycosylation and stability and resulted in the retention of the enzyme in the endoplasmic reticulum. These phenotypes were partially complemented by expression of the proregion in trans. Trans complementation was specific to Kex2p proregion because expression of any of the seven mammalian prohormone convertase propeptides had no effect. These data are consistent with a model whereby Kex2p proregion functions as an intramolecular chaperone and indicate that covalent linkage to the protein is not an absolute requirement for proregion function. Furthermore, extensive mutagenesis revealed that, in addition to their function as proteolytic recognition sites, C-terminal basic residues play an active role in proregion-dependent Kex2p activation.

Low specificity of 2 tetanus rapid tests in Cambodia.

OBJECTIVES: Rapid testing for tetanus on serum or blood allows for an immediate evaluation of individual protection against tetanus in developed countries, using a "single step" immunochromatographic technique using tetanus toxoid. The specificity of these tests, compared to the reference method for tetanus, mouse serum neutralization testing, has however never been assessed in these countries, due to the difficulty to perform serum neutralization titration in mice, because of animal testing bioethical regulations. POPULATION AND METHODS: A collection of sera from adult volunteers in Cambodia, living in rural environment, was tested for tetanus antibodies by ELISA in France, and by mouse serum neutralization in Vietnam. This allowed estimating the sensitivity and specificity of 2 rapid tetanus tests, available on the market: TQS™ and Tetanotop™. RESULTS: The sensitivity of these tests was adequate, compared to mice serum neutralization test, for a test threshold of 0.01 IU/mL, (100% for TQS™, 91% for Tetanotop™), but their specificity was very low (1% for TQS™ and 13% for Tetanotop™). CONCLUSION: The results prove that these rapid tests for the assessment of individual protection against tetanus should not be used in the adult rural Cambodian population.

Mechanism of Kex2p inhibition by its proregion.

Many proteases are produced as zymogens bearing an N-terminal proregion acting both as intramolecular chaperone and as enzyme inhibitor. We studied here the inhibition mechanism of the yeast proprotein convertase Kex2p by its proregion. A recombinant secreted and soluble form of Kex2p was produced in Pichia pastoris and its enzymatic properties toward a fluorogenic synthetic peptide were characterized. Recombinant Escherichia coli-produced Kex2p proregion specifically and potently inhibited the enzyme, with an IC(50) of 160 nM. Exploration of the inhibition mechanism revealed that the proregion behaved as a mixed inhibitor.

Cystadenoma of the proximal common hepatic duct: the use of abdominal ultrasonography and transhepatic cholangiography in diagnosis.

Cystadenoma of the extrahepatic bile ducts is a rare cause of obstructive jaundice. In a case seen recently at our institution, the combined findings are abdominal ultrasonography and transhepatic cholangiography were diagnostic; such studies should provide evidence for preoperative recognition.

Incidence of hepatitis C virus infection in burn patients: detection of anti-C100, anti-C33c and anti-Core antibodies.

Anti-hepatitis C virus antibodies were searched for in 45 burnt patients, at the time of burn injury and more than 6 months after burn injury. HCV infection was detected in 18% as a consequence of the numerous transfusions of blood or blood derivatives used during the post-burn treatment. Five patients displayed evidence of anti-C100, anti-C33c and anti-Core antibodies together; two patients had only anti-C100 and anti-C33c antibodies, and the last one showed only anti-Core antibodies. Chronic hepatitis was observed in 83% of HCV infections. Kinetics of appearance of anti-HCV antibodies varied between patients. Anti-Core is generally the first to be detected at high levels; however, in at least one case it was detected only two and a half months after C100 and C33c antibodies.

Dynamic modeling of biodegradation and volatilization of hazardous aromatic substances in aerobic bioreactor.

The aerobic biological process is one of the best technologies available for removing hazardous organic substances from industrial wastewaters. But in the case of volatile organic compounds (benzene, toluene, ethylbenzene, p-xylene, naphthalene), volatilization can contribute significantly to their removal from the liquid phase. One major issue is to predict the competition between volatilization and biodegradation in biological process depending on the target molecule. The aim of this study was to develop an integrated dynamic model to evaluate the influence of operating conditions, kinetic parameters and physical properties of the molecule on the main pathways (biodegradation and volatilization) for the removal of Volatile Organic Compounds (VOC). After a comparison with experimental data, sensitivity studies were carried out in order to optimize the aerated biological process. Acclimatized biomass growth is limited by volatilization, which reduces the bioavailability of the substrate. Moreover, the amount of biodegraded substrate is directly proportional to the amount of active biomass stabilized in the process. Model outputs predict that biodegradation is enhanced at high SRT for molecules with low H and with a high growth rate population. Air flow rate should be optimized to meet the oxygen demand and to minimize VOC stripping. Finally, the feeding strategy was found to be the most influential operating parameter that should be adjusted in order to enhance VOC biodegradation and to limit their volatilization in sequencing batch reactors (SBR).

Regulation of cholangiocyte proliferation.

Intrahepatic bile duct epithelial cells (i.e., cholangiocytes) are the target cells of chronic cholestatic liver diseases (i.e., cholangiopathies), which makes these cells of great interest to clinical hepatologists. This review will focus on "typical" cholangiocyte proliferation, whereas "atypical" (extension of cholangiocyte proliferation into parenchyma), and premalignant "oval" cell proliferation are reviewed elsewhere. The bile duct ligated (BDL) rat model, where most of the known mechanisms of cholangiocyte proliferation have been illustrated, was the first and remains the prototype animal model for "typical" cholangiocyte proliferation. Following a short overview of cholangiocyte functions, we briefly discuss the: (i) in vivo models [i.e., BDL (Fig. 1 and 4), chronic alpha-naphthylisothiocyanate (ANIT) or bile acid feeding (Fig. 2), acute carbon tetrachloride (CCl4) feeding and partial hepatectomy; and (ii) in vitro experimental tools [e.g., purified cholangiocytes and isolated intrahepatic bile duct units (IBDU)] that are key to the understanding of the mechanisms of "typical" cholangiocyte growth. In the second part of the review, we discuss a number of potential factors or conditions [e.g., gastrointestinal hormones, nerves, estrogens, blood supply, and growth factors] as well as the intracellular mechanisms [e.g., adenosine 3',5'-monophosphate (cAMP), and protein kinase C (PKC)] that may regulate "typical" cholangiocyte hyperplasia.

Bile acid-dependent vesicular transport of lysosomal enzymes into bile in the rat.

BACKGROUND: Bile acids may stimulate the movement of hepatocyte vesicles and enhance their fusion with the biliary canaliculus. The present study examined the effects of various bile acids on the exocytosis of the contents of hepatocyte lysosomes into the biliary canaliculus. METHODS: The effects of various bile acids on hepatocyte lysosome movement and on exocytosis of the contents of hepatocyte lysosomes into the biliary canaliculus were determined from the distribution of fluorescein isothiocyanate-dextran--labeled lysosomes in hepatocyte couplets and by quantitating biliary lysosomal enzyme output in rats. RESULTS: Hydrophobic as well as hydrophilic and nonmicellar bile acids were found to stimulate to a similar degree the output of lysosomal enzymes into bile, indicating that bile acid-induced change of canalicular or lysosomal membrane fluidity is not responsible for enhanced exocytosis. The taurocholate-dependent increase in lysosomal enzyme excretion was completely blocked by either microtubule or microfilament inhibition, suggesting that these subcellular structures are involved in bile acid-dependent vesicular transport. Fluorescent microscopy studies showed that taurocholate causes a microtubule-dependent translocation of lysosomes towards the canaliculus in hepatocyte couplets, which occurred at the same time as increased output of lysosomal enzymes into bile. CONCLUSIONS: The results suggest that bile acids modulate vesicle traffic towards the canaliculus by a mechanism unrelated to bile acid interaction with the vesicle membrane.

Demonstration of vesicular-dependent bile flow in the sucrose-loaded rat.

The contribution of the hepatocyte vacuolar apparatus to bile fluid formation was assessed by studying the transcellular transport and biliary excretion of the fluid-phase marker sucrose. In rats sucrose-loaded by IP administration of sucrose, electron microscopy showed expansion of the vacuolar apparatus and numerous large lysosomelike structures in hepatocytes. Subcellular distribution studies showed that sucrose was sequestered in lysosomes. Compared with controls, sucrose-loaded rats had a 30% higher (P less than 0.01) bile flow with no change in biliary bile acid or electrolyte concentrations. Administration of colchicine ablated the sucrose-induced choleresis and resulted in parallel changes in biliary secretions of sucrose and lysosomal enzymes. Our data suggest that in the sucrose-loaded rat, the hepatocyte vacuolar apparatus may contribute significantly to bile formation by microtubule-dependent release of fluid into bile by exocytosis.

Regulation of cholangiocyte bicarbonate secretion.

The objective of this review article is to discuss the role of secretin and its receptor in the regulation of the secretory activity of intrahepatic bile duct epithelial cells (i.e., cholangiocytes). After a brief overview of cholangiocyte functions, we provide an historical background for the role of secretin and its receptor in the regulation of ductal secretion. We review the newly developed experimental in vivo and in vitro tools, which lead to understanding of the mechanisms of secretin regulation of cholangiocyte functions. After a description of the intracellular mechanisms by which secretin stimulates ductal secretion, we discuss the heterogeneous responses of different-sized intrahepatic bile ducts to gastrointestinal hormones. Furthermore, we outline the role of a number of cooperative factors (e.g., nerves, alkaline phosphatase, gastrointestinal hormones, neuropeptides, and bile acids) in the regulation of secretin-stimulated ductal secretion. Finally, we discuss other factors that may also play an important role in the regulation of secretin-stimulated ductal secretion.

Purification and immunological quantification of rat liver lysosomal glycosidases.

Although lysosomal enzyme activities are known to vary in response to numerous physiological and pharmacological stimuli, the relationship between lysosomal enzyme activity and enzyme concentration has not been systematically studied. Therefore we developed radioimmunoassays for two lysosomal glycosidases in order to determine lysosomal enzyme concentration. beta-Galactosidase and beta-glucuronidase were purified from rat liver 2780-fold and 1280-fold respectively, by using differential centrifugation, affinity chromatography, ion-exchange chromatography and molecular-sieve chromatography. Polyclonal antibodies to these enzymes were raised in rabbits, and two radioimmunoassays were established. Antibody specificity was shown by: (i) selective immunoprecipitation of enzyme activity; (ii) identical bands of purified enzyme on SDS/polyacrylamide-gel electrophoresis and immunoelectrophoresis; (iii) single immunoreactive peaks in molecular-sieve chromatography experiments. Sensitivities of the assays were such that 15 ng of beta-galactosidase and 45 ng of beta-glucuronidase decreased the ratio of bound to free radiolabel by 50%; minimal detectable amounts of immunoreactive enzymes were 2 ng and 10 ng respectively. The assays were initially used to assess the effects of physiological perturbations (i.e. fasting and age) on enzyme concentrations in rat liver; these experiments showed that changes in enzyme concentrations do not always correlate with changes in enzyme activities. This represents the first report of radioimmunoassays for lysosomal glycosidases. The results suggest that these radioimmunoassays provide useful technology for the study of regulatory control mechanisms of the concentrations of lysosomal glycosidases in mammalian tissues.

[Detection of IgM anti-hepatitis A antibodies by immunoenzyme technic. Interpretation of results and comparison of 2 tests (Havab-M EIA and Hepanostika anti-HAV IgM]. / Détection des anticorps anti-hépatite A de classe IgM par méthode immunoenzymatique. Interprétation des résultats et comparaison de deux tests (Havab-M EIA, et Hepanostika anti-HAV IgM).

The presence of IgM anti-HAV was investigated in 60 patients with acute hepatitis A using two commercialy available enzyme linked immunosorbent assays (Havab-M EIA, Abbott, and Hepanostika anti-HAV IgM, Organon). High levels of anti-HAV IgM were present for only 30 to 60 days after the onset. Low levels of anti-HAV IgM were detected for as long as two years after acute phase. These results suggest that only high levels of anti-HAV IgM have to be considered for the diagnosis of a recent infection by hepatitis A virus.