Abiotic transformation of catechol and 1-naphthol in aqueous solution-influence of environmental factors.

The abiotic transformation of catechol and 1-naphthol singly and in mixtures was tested in sterile Tris-HCl buffer with regard to several environmental factors including temperature (7 degrees C, 20 degrees C and 30 degrees C), lighting conditions, pH (between 7.0 and 8.5) and dissolved oxygen (at partial pressures of 0.0, 220, 2200, 11000 and 22000 Pa). Irrespective of lighting conditions. catechol autoxidation was confirmed in aerated medium with a rate independent of the presence of 1-naphthol but proportional to the dissolved oxygen concentration, to the pH (its half-disappearance occurred in 24h at pH 8.5) and, to a lesser extent, to the incubating temperature (at 20 degrees C, 20% disappeared in 10 days at pH 7.0). Under alkaline conditions, the reaction of the anionic form (catecholate) with an equimolar concentration of molecular oxygen (O2) led presumably to hydrogen peroxide anion (HO2-) and coloured polymerization products. When tested alone, 1-naphthol was not significantly influenced either by lighting conditions, incubating temperature or dissolved oxygen concentration. It was also found to be quite stable with respect to pH, with a 15-fold weaker transformation rate than for catechol at the highest pH used. When tested in a mixture with catechol, 1-naphthol was found to be involved in a new chemical oxidation reaction catalyzed by catecholate. The transformation of one mole of 1-naphthol consumes four moles of oxygen. In the presence of catechol, the stoichiometry of the 1-naphthol transformation, under the influence of oxygen, suggests the possible formation of 2,5,6,8-tetrahydroxy 1,4-naphthoquinone via Lawsone (2-hydroxy 1,4-naphthoquinone) and naphthopurpurine (2,5,8-trihydroxy 1,4-naphthoquinone) as hypothetic intermediates. This is the first report of the autoxidation of 1-naphthol, catalyzed by catechol, in aqueous solution, in the absence of UV irradiation.

Pressure-drops control strategy in a fixed-bed reactor.

This paper presents a strategy to control pressure-drops (head loss) in a biofilter designed according to the "Mist-Foam" concept. This concept is based on the mixing of the gaseous substrate and a liquid nutrient solution with an atomization nozzle to generate a mist passing subsequently through a synthetic polyurethane foam. In this type of bioreactor, the microbial growth reduces progressively the empty bed volume of the biofilter and causes an increase in the pressure-drops. This phenomenon can result in a complete clogging of the biofilter. The strategy of pressure-drops control presented here consists of successive interruption of the liquid flow, automatically controlled, resulting in a drying effect of the biomass. Tested during a 160 days experiment, this system has permitted to reduce and stabilize the pressure-drops in a biofilter in which the carrier exhibited a high likelihood of clogging.

Glutathione-mediated mineralization of 14C-labeled 2-amino-4,6-dinitrotoluene by manganese-dependent peroxidase H5 from the white-rot fungus Phanerochaete chrysosporium.

Manganese-dependent peroxidase (MnP) H5 from the white-rot fungus Phanerochaete chrysosporium, in the presence of either Mn(II) (10 mM) or GSH (10 mM). was able to mineralize 14C-U-ring-labeled 2-amino-4,6-dinitrotoluene (2-A-4,6-DNT) up to 29% in 12 days. When both Mn(II) and GSH were present, the mineralization extent reached 82%. On the other hand, no significant mineralization was observed in the absence of both Mn(II) and GSH, suggesting the requirement of a mediator [either Mn(II) or GSH] for the degradation of 2-A-4,6-DNT by MnP. Using electron spin resonance (ESR) techniques, it was found that the glutathionyl free radical (GS*) was produced through the oxidation of GSH by MnP in the presence as well as in the absence of Mn(II). GS* was also generated through the direct oxidation of GSH by Mn(III). Our results strongly suggest the involvement of GS* in the GSH-mediated mineralization of 2-A-4,6-DNT by MnP.

ATR-FTIR sensor development for continuous on-line monitoring of chlorinated aliphatic hydrocarbons in a fixed-bed bioreactor.

This article describes the continuous on-line monitoring of a dechlorination process by a novel attenuated total reflection-Fourier transform infrared (ATR-FTIR) sensor. This optical sensor was developed to measure noninvasively part-per-million (ppm) concentrations of trichloroethylene (TCE), tetrachloroethylene (PCE), and carbon tetrachloride (CT) in the aqueous effluent of a fixed-bed dechlorinating bioreactor, without any prior sample preparation. The sensor was based on an ATR internal reflection element (IRE) coated with an extracting hydrophobic polymer, which prevented water molecules from interacting with the infrared (IR) radiation. The selective diffusion of chlorinated compound molecules from aqueous solution into the polymer made possible their detection by the IR beam. With the exclusion of water the detection limits were lowered, and measurements in the low ppm level became possible. The best extracting polymer was polyisobutylene (PIB) in the form of a 5.8-microm thick film, which afforded a detection limit of 2, 3, and 2. 5 mg/L (ppm) for TCE, PCE, and CT, respectively. Values of the enrichment factors between the polymer coating and the water matrix of these chloro-organics were determined experimentally and were compared individually with predictions obtained from the slopes of absorbance/concentration curves for the three analytes. Before coupling the ATR-FTIR sensor to the dechlorinating bioreactor, preliminary spectra of the chlorinated compounds were acquired on a laboratory scale configuration in stop-flow and flow-through closed-loop modes. In this way, it was possible to study the behavior and direct response of the optical sensor to any arbitrary concentration change of the analytes. Subsequently, the bioreactor was monitored with the infrared sensor coupled permanently to it. The sensor tracked the progression of the analytes' spectra over time without perturbing the dechlorinating process. To calibrate the ATR-FTIR sensor, a total of 13 standard mixtures of TCE, PCE and CT at concentrations ranging from 0 to 60 ppm were selected according to a closed symmetrical experimental design derived from a 3(2) full-factorial design. The above range of concentrations chosen for calibration reflected typical values during normal bioreactor operation. Several partial least squares (PLS) calibration models were generated to resolve overlapping absorption bands. The standard error of prediction (SEP) ranged between 0.6 and 1 ppm, with a relative standard error of prediction (RSEP) between 3 and 6% for the three analytes. The accuracy of this ATR-FTIR sensor was checked against gas chromatography (GC) measurements of the chlorocompounds in the bioreactor effluents. The results demonstrate the efficiency of this new sensor for routine continuous on-line monitoring of the dechlorinating bioreactor. This strategy is promising for bioprocess control and optimization.

Continuous degradation of mixtures of 4-nitrobenzoate and 4-aminobenzoate by immobilized cells of Burkholderia cepacia strain PB4.

Although isolated on 4-aminobenzoate, Burkholderia cepacia strain PB4 is also able to grow on 4-nitrobenzoate. Degradation of an equimolar mixture of the nitroaromatic compound 4-nitrobenzoate and its corresponding aminoaromatic derivative 4-aminobenzoate by this strain was investigated. Batch experiments showed that, irrespective of preculturing conditions, both compounds were degraded simultaneously. The mixture-degrading ability of B. cepacia strain PB4 was subsequently tested in continuous packed bed reactors (PBR) with the strain immobilized on Celite grade R-633 or R-635. Higher degradation rates were achieved with the larger particles of Celite R-635. Maximum simultaneous degradation rates per liter of packed bed of 0.925 mmol 1(-1) h(-1) 4-nitrobenzoate and 4-aminobenzoate were obtained for an applied loading rate of the same value (0.925 mmol 1(-1) h(-1) of each compound). Even when the applied load was not removed in its entirety, neither of the two compounds was degraded preferentially but a percentage of both of them was mineralized. The present study shows the possibility for a pure strain to biodegrade not only a nitroaromatic compound (4-nitrobenzoate) but also its corresponding amino derivative (4-aminobenzoate) continuously and simultaneously.

Transformation and mineralization of 2,4,6-trinitrotoluene (TNT) by manganese peroxidase from the white-rot basidiomycete Phlebia radiata.

The degradation of the nitroaromatic pollutant 2,4,6-trinitrotoluene (TNT) by the manganese-dependent peroxidase (MnP) of the white-rot fungus Phlebia radiata and the main reduction products formed were investigated. In the presence of small amounts of reduced glutathione (10 mM), a concentrated cell-free preparation of MnP from P. radiata exhibiting an activity of 36 nkat/ml (36 nmol Mn(II) oxidized per sec and per ml) transformed 10 mg/l of TNT within three days. The same preparation was capable of completely transforming the reduced derivatives of TNT. When present at 10 mg/l, the aminodinitrotoluenes were transformed in less than two days and the diaminonitrotoluenes in less than three hours. Experiments with 14C-U-ring labeled TNT and 2-amino-4,6-dinitrotoluene showed that these compounds were mineralized by 22% and 76%, respectively, within 5 days. Higher concentrations of reduced glutathione (50 mM) led to a severe inhibition of the degradation process. It is concluded that Phlebia radiata is a good candidate for the biodegradation of TNT as well as its reduction metabolites.

Anaerobic dechlorinating bacteria.

Anaerobic dehalogenation is attracting great interest since it opens new research horizons based on the novel biochemical mechanisms identified in this field such as halorespiration, i.e. the utilization of halogenated compounds as electron acceptors. Moreover, anaerobic bacteria seem to be more efficient than their aerobic counterparts in removing halogen atoms from polyhalogenated compounds. Thus, anaerobic dehalogenation can be considered as a promising means for bioremediation treatments of persistently polluted environments. In this line, identification of pure strains capable of dehalogenation will give important information about the diversity of organisms implicated in this process and also fundamental explanations of the diverse biochemical mechanisms involved. In light of these considerations, we chose to focus this review on the physiological descriptions, dechlorination activities, phylogenetic diversity, and potential biotechnological applications of these pure anaerobic strains capable of dehalogenation.

Effect of specific inhibitors on the anaerobic reductive dechlorination of 2,4,6-trichlorophenol by a stable methanogenic consortium.

The transformation of 2,4,6-trichlorophenol (TCP) into 4-chlorophenol (4CP) was studied using a stable methanogenic enrichment culture derived from an anaerobic fixed bed reactor. Using acetate as a growth substrate, different inhibitors of methanogenesis exhibited distinct effects on TCP dechlorination. Whereas reductive dechlorination activity was not affected by 2% ethylene in the gas phase, 25 mM bromoethanesulfonic acid (BESA) had a direct inhibitory effect on this process. The choice of BESA as a specific inhibitor for identifying the subpopulations involved in reductive dechlorination of chloroaromatics is thus questionable. Inhibitors of sulfate reduction such as molybdate (20 mM) and selenate (20 mM) had a direct inhibitory effect on reductive dechlorination independently of the presence of sulfate in the medium supplemented with acetate as growth substrate. Consequently much more care must also be taken with these inhibitors to prove that reductive chlorination is coupled to sulfate reduction.

Introduction of anaerobic dechlorinating bacteria into soil slurry microcosms and nested-PCR monitoring.

Desulfomonile tiedjei and Desulfitobacterium dehalogenans were chosen as model bacteria to demonstrate the introduction of an anaerobic microbia reductive dechlorination activity into nonsterile soil slurry microcosms by inoculation. De novo 3-chlorobenzoate dechlorination activity was established with the bacterium D. tiedjei in microcosms normally devoid of this dechlorination capacity. The addition of D. tiedjei to microcosms supplemented with 20 mM pyruvate as the cosubstrate resulted in total biotransformation of 1.5 mM 3-chlorobenzoate within 7 days. The introduction of the bacterium Desulfitobacterium dehalogenans into nonsterile microcosms resulted in a shortening of the period required for dechlorination activity to be established. In microcosms inoculated with Desulfitobacterium dehalogenans, total degradation of 6 mM 3-chloro-4-hydroxy phenoxyacetic acid (3-Cl-4-OHPA) was observed after 4 days in contrast to the result in noninoculated microcosms, where the total degradation of 3-Cl-4-OHPA by indigenous microorganisms was observed after 11 days. Both externally introduced bacterial strains were detected in soil slurry microcosms by a nested-PCR methodology.

Introduction and PCR detection of Desulfomonile tiedjei in soil slurry microcosms.

The aim of this work was to test the feasibility of introducing an anaerobic microbial reductive dechlorination activity into non sterile soil slurry microcosms by inoculation with the pure anaerobic bacterial strain Desulfomonile tiedjei, which is capable of dechlorinating 3-chlorobenzoate to benzoate. To show that the bacterium was established in the microcosms we followed the expression of the reductive dechlorination activity and a molecular probe based on PCR amplification of the 16S rDNA gene was developed. However, the success of PCR amplification of the 16S rDNA gene depends on the DNA extraction and purification methodologies applied, as shown through the use of several protocols. In this study we report a DNA extraction and purification method which generates sufficient and very clean DNA suitable for PCR amplification of the D. tiedjei 16S rDNA gene. The threshold of detection was about 5.10(3) bacteria per gram of soil slurry. Introduction of D. tiedjei in soil slurry microcosms proved successful since 3-chlorobenzoate dechlorination activity was established with this bacterium in microcosms normally devoid of this dechlorination capacity. Indeed, the addition of D. tiedjei to microcosms supplemented with acetate plus formate as cosubstrate, at their respective concentrations of 5 and 6 mM, led to a total biotransformation of 2.5 mM of 3-chlorobenzoate within 12 days. After complete 3-chlorobenzoate dechlorination, the 16S rDNA gene of this bacterium was specifically detected only in the inoculated microcosms as shown by PCR amplification followed by restriction mapping confirmation.

High-rate continuous biodegradation of concentrated chlorinated aliphatics by a durable enrichment of methanogenic origin under carrier-dependent conditions.

The simultaneous biodegradation of toxic compounds in mixtures is a major current concern. To bioremediate a toxic mixture, we designed a strategy combining an ad-sorbent carrier with an ecological and nutritional system which allowed work close to heavily polluted conditions in nature. Starting from a methanogenic community, we developed a microbial consortium acclimated to a mixture of about 30 chlorinated aliphatics in a fixed-film stationary-bed bioreactor. Prior to the establishment of a durable period of dechlorination, an interval of progressive dechlorination of the toxic mixture was observed during which the excess of the toxic compounds was stored on the carrier. The latter, consisting of activated carbon in a polyurethane foam, allowed us to work at concentrations far above the solubility of the toxic compounds (apparent concentrations of about 10 g/L). The complete disappearance of hexachloroethane as well as its lower homologues, penta-, tetra-, and trichloroethane, present in the toxic mixture, was observed. Additionally, octachlorocyclopentene, carbon tetrachloride, trichloro-ethylene, tetrachloroethylene, and hexachloro-1,3-butadiene also completely disappeared. For the four latter compounds, from mass balances in the bioreactor, degradation rates around 10 mumol/L per day were determined with total dechlorination. The enrichment culture thus developed exhibited high degradation performances similar to those reported in the literature for pure or enriched anaerobic microbial cultures in contact with a single toxic compound. The results demonstrate the possibility of concurrent high-rate degradation of several highly chlorinated toxic compounds, under conditions approximating field situations.(c) 1995 John Wiley & Sons, Inc.

Computation of pH evolution versus ionic products concentration in a fermentation broth.

An algorithm developed for pH computation has been tested to calculate the theoretical pH changes in a culture medium during the course of a fermentation. A divergence between the computed pH value and the value measured with the electrode allows us to highlight the presence of undetected ionic products. The calculation with the algorithm by means of a computer requires only the knowledge of the ionic properties of the substrates and detected products and existing thermodynamic constants.

Implementation of an adaptive controller for the startup and steady-state running of a biomethanation process operated in the CSTR mode.

An adaptive control algorithm has been implemented on a biomethanation process to maintain propionate concentration, a stable variable, at a given low value, by steering the dilution rate. It was thereby expected to ensure the stability of the process during the startup and during steady-state running with an acceptable performance. The methane pilot reactor was operated in the completely mixed, once-through mode and computer-controlled during 161 days. The results yielded the real-life validation of the adaptive control algorithm, and documented the stability and acceptable performance expected.

[Removal of an anatomic post]. / Dépose du tenon anatomique.

Removing an anatomical cast post requires many precaution and, in most situation, gives very good results. Nevertheless, this procedure can be dangerous for the tooth or the surrounding tissues and must be considered only in case of absolute necessity. Two techniques are described using a little hole made in the coronal part of the core. A threaded wire is driven through this hole making possible the use of a crown remover. For the second technique the use of an original appliance, the ATD bridge remover, is demonstrated with very good results too.

Trophic relationships between Saccharomyces cerevisiae and Lactobacillus plantarum and their metabolism of glucose and citrate.

Glucose and citrate are two major carbon sources in fruits or fruit juices such as orange juice. Their metabolism and the microorganisms involved in their degradation were studied by inoculating with an aliquot of fermented orange juice a synthetic model medium containing glucose and citrate. At pH 3.6, their degradation led, first, to the formation of ethanol due to the activity of yeasts fermenting glucose and, eventually, to the formation of acetate resulting from the activity of lactobacilli. The yeast population always outcompeted the lactobacilli even when the fermented orange juice used as inoculum was mixed with fermented beet leaves containing a wider variety of lactic acid bacteria. The evolution of the medium remained similar between pH 3.3 and 5.0. At pH 3.0 or below, the fermentation of citrate was totally inhibited. Saccharomyces cerevisiae and Lactobacillus plantarum were identified as the only dominant microorganisms. The evolution of the model medium with the complex microbial community was successfully reconstituted with a defined coculture of S. cerevisiae and L. plantarum. The study of the fermentation of the defined model medium with a reconstituted microbial community allows us to better understand the behavior not only of fermented orange juice but also of many other fruit fermentations utilized for the production of alcoholic beverages.

Fermentation of citrate by Lactobacillus plantarum in the presence of a yeast under acid conditions.

At pH 3.6, Lactobacillus plantarum is unable to grow on citrate or to ferment it in the absence of another carbon source such as glucose. In a defined medium containing glucose and citrate, with a higher concentration of the former than the latter, as in many fermented alcoholic beverages, L. plantarum will first ferment the sugar. The production of lactate from glucose degradation increases the acidity of the medium and inhibits the fermentation of citrate. In co-culture with Saccharomyces cerevisiae, part of the glucose is fermented by the yeast, partly avoiding the pH drop and the inhibition of citrate fermentation by L. plantarum. Fermentation was still possible at pH values around 3.0.

Transient response of a solid-liquid model biological fluidised bed to a step change in fluid superficial velocity.

The evolution of a solid-liquid model biological fluidised bed under a step change in fluid superficial velocity is described. During a transient step change, the fluidised bed divides into a top zone which remains at the initial porosity and a bottom zone which settles at the final porosity. The interface of discontinuity in porosity moves progressively upwards through the fluidised bed. The velocity at which the top of the fluidised bed expands or contracts and the upward velocity of the porosity transition interface depend only upon the initial and final states of the bed porosity and the fluid superficial velocity. This results in a linear evolution with time of the total bed height and the height of porosity transition interface. The proposed model is well suited to describe the transient response of low-density particles in a fluidised bed, such as encountered in biological systems, to a sudden change of liquid superficial velocity. The model was validated experimentally.

[Removal of fixed prosthetics]. / Dépose des éléments prothétiques fixés.

Unscaling a fixed prosthesis can be dangerous for the supporting tissue. However a careful examination of the clinical situation and the proper use of correct technics described in the following article, may prudence a successful result most of the time.

Computation of physicochemical parameters, namely pH, in complex (bio)chemical systems: extension to gas-liquid systems.

The first model has been proposed to compute, in complex liquid (bio)chemical systems, a number of physicochemical parameters, namely pH, concentration of one of any chemical species, partition between acid-base forms, global charge, or ionic strength, assuming the physicochemical equilibrium state. The extension of the present model, described here, permits moreover the computation of gas-liquid distributions, specific gas volumes, or total pressures. The model solely requires the knowledge of existing thermodynamic constants and of the concentration of every chemical species other than the species under examination. The model elicits a unique equilibrium state. Computed values agreed with experimental measurements, thereby validating the model. Digital computer programs were prepared to use the proposed algorithms.

Problematics and stability of on-line pH measurements in anaerobic environments: The jellied combined electrode.

Discrepancies of one pH unit and more have been observed after a few days, between continuous on-line in situ pH measurements and instant off-line pH measurements during anaerobic digestion of an agroindustrial wastewater. Concomitantly, the electrical resistance across the porous diaphragm of the on-line electrode increased, and a black clogging developed on its diaphragm. Measurements of the relative liquid junction potential in KCl or Na(2)S solutions excluded that high concentrations of ions such as, K(+), Na(+), Cl(-), HS(-), or S(2-) were the major cause of the drifts in pH values. It has been possible to limit the rapid increase of the liquid junction potential and the black clogging formation on the porous diaphragm either by acidification and/or by overpressurization of the electrode-filling liquid. Continuous on-line in situ pH values consistent with instant off-line pH values over long periods of time have been obtained with a newly designed pH sensor in which a special jellied electrode filling replaced the porous diaphragm.