Enhancement of the denitrification performance of an activated sludge using an electromagnetic field in batch mode.

The influence of electromagnetic fields on bacterial denitrification has been tested on synthetic media with sludges from wastewater treatment stations, in batch mode. The effects of the intensity of the magnetic induction ratio B (mT), reaction volume and initial biomass concentration on the kinetics of the denitrification process were studied. Magnetic field had both an optimal stimulating effect on the activity of the denitrifying flora for B (mT)/mgx values of the order of 0.212, and an inhibitory effect for the values beyond the latter.Sludges underwent multiple exposure cycles to magnetic fields. It was shown that, after three exposure cycles, denitrification kinetics went from 6.5 to 12.7 mg N-NO-3.L-1.h-1 which corresponds to a 2.7 fold improvement. The improved performance persists even after the cessation of the magnetic field. Observation of the sludge by the environmentalelectron microscope shows that the microbial population forming the starting sludge; changed following exposure to the magnetic field. The action of the; electromagnetic field on the microbial populations in denitrification resulted in the modification of the diversity of the flora that is initially present, favoring the development of Proteo bacteria, particularly the Betaproteo bacteria subclass, which results in improved denitrification.

Solid anaerobic digestion: State-of-art, scientific and technological hurdles.

In this paper, a state-of-art about solid anaerobic digestion (AD), focused on recent progress and trends of research is proposed. Solid anaerobic digestion should be the most appropriate process for degradation of by-products with high total solid (TS) content, especially lignocellulosic materials like agricultural waste (straw, manure), household waste and food waste. Solid AD is already widely used in waste water treatment plant for treating plant for sewage sludge but could be more developed for lignocellulosic materials with high TS content. Many research works were carried out in Europe on solid AD, focused on current hurdles (BMP, codigestion, inhibition, microbial population, rheology, water transfers, inoculum, etc.) in order to optimize the solid AD process. In conclusion, hurdles of solid AD process should and must be solved in order to propose better productivity and profitability of such system operating with high TS content (>15%), favouring reliable industrial processes.

DLVO, hydrophobic, capillary and hydrodynamic forces acting on bacteria at solid-air-water interfaces: Their relative impact on bacteria deposition mechanisms in unsaturated porous media.

Experimental and modeling studies were performed to investigate bacteria deposition behavior in unsaturated porous media. The coupled effect of different forces, acting on bacteria at solid-air-water interfaces and their relative importance on bacteria deposition mechanisms was explored by calculating Derjaguin-Landau-Verwey-Overbeek (DLVO) and non-DLVO interactions such as hydrophobic, capillary and hydrodynamic forces. Negatively charged non-motile bacteria and quartz sands were used in packed column experiments. The breakthrough curves and retention profiles of bacteria were simulated using the modified Mobile-IMmobile (MIM) model, to identify physico-chemical attachment or physical straining mechanisms involved in bacteria retention. These results indicated that both mechanisms might occur in both sand. However, the attachment was found to be a reversible process, because attachment coefficients were similar to those of detachment. DLVO calculations supported these results: the primary minimum did not exist, suggesting no permanent retention of bacteria to solid-water and air-water interfaces. Calculated hydrodynamic and resisting torques predicted that bacteria detachment in the secondary minimum might occur. The capillary potential energy was greater than DLVO, hydrophobic and hydrodynamic potential energies, suggesting that film straining by capillary forces might largely govern bacteria deposition under unsaturated conditions.

Towards a standardization of biomethane potential tests.

Production of biogas from different organic materials is a most interesting source of renewable energy. The biomethane potential (BMP) of these materials has to be determined to get insight in design parameters for anaerobic digesters. Although several norms and guidelines for BMP tests exist, inter-laboratory tests regularly show high variability of BMPs for the same substrate. A workshop was held in June 2015, in Leysin, Switzerland, with over 40 attendees from 30 laboratories around the world, to agree on common solutions to the conundrum of inconsistent BMP test results. This paper presents the consensus of the intense roundtable discussions and cross-comparison of methodologies used in respective laboratories. Compulsory elements for the validation of BMP results were defined. They include the minimal number of replicates, the request to carry out blank and positive control assays, a criterion for the test duration, details on BMP calculation, and last but not least criteria for rejection of the BMP tests. Finally, recommendations on items that strongly influence the outcome of BMP tests such as inoculum characteristics, substrate preparation, test setup, and data analysis are presented to increase the probability of obtaining validated and reproducible results.

Early assessment of a rapid alternative method for the estimation of the biomethane potential of sewage sludge.

This short communication briefly presents a rapid method using a fluorescent redox indicator, similar to resazurin, in order to estimate the biodegradability of sewage sludge during anaerobic digestion (AD). The biodegradability and by extension the Biochemical Methane Potential (BMP) of nineteen municipal sludge samples (primary, biological and tertiary) were investigated and estimated in only 48 h. Results showed the relevance to follow the metabolic activity of anaerobic sludge by the kinetic of probe reduction. The extended lag phase of inoculum indicated an impact of pre-treatments on enzyme activity. The comparison with Automatic Methane Potential Test System II (AMPTS) confirmed the estimated values of BMP according to an uncertainty limit of 25%. These first results highlight the interest of this rapid assay as a preliminary tool of the biodegradability of sewage sludge in anaerobic digestion.

Bacteria cell properties and grain size impact on bacteria transport and deposition in porous media.

The simultaneous role of bacteria cell properties and porous media grain size on bacteria transport and deposition behavior was investigated in this study. Transport column experiments and numerical HYDRUS-1D simulations of three bacteria with different cell properties (Escherichia coli, Klebsiella oxytoca, and Rhodococcus rhodochrous) were carried out on two sandy media with different grain sizes, under saturated steady state flow conditions. Each bacterium was characterized by cell size and shape, cell motility, electrophoretic mobility, zeta potential, hydrophobicity and potential of interaction with the sand surface. Cell characteristics affected bacteria transport behavior in the fine sand, but similar bacteria breakthroughs and retardation factors observed in the coarse sand, indicated that bacteria transport was more depended on grain size than on bacteria cell properties. Retention decreased with increasing hydrophobicity and increased with increasing electrophoretic mobility of bacteria for both sand. The increasing sand grain size resulted in a decrease of bacteria retention, except for the motile E. coli, indicating that retention of this strain was more dependent on cell motility than on the sand grain size. Bacteria deposition coefficients obtained from numerical simulations of the retention profiles indicated that straining was an important mechanism affecting bacteria deposition of E. coli and Klebsiella sp., in the fine sand, but the attachment had the same importance as straining for R. rhodochrous. The results obtained in the coarse sand did not permit to discriminate the predominant mechanism of bacteria deposition and the relative implication of bacteria cell properties of this process.

Nitrite accumulation during denitrification depends on the carbon quality and quantity in wastewater treatment with biofilters.

This study aims to understand the mechanisms of nitrite appearance during wastewater denitrification by biofilters, focusing on the role of the carbon source. Experiments were carried out at lab-scale (batch tests) and full-scale plant (Parisian plant, capacities of 240,000 m(3) day(-1)). Results showed that the nature of the carbon source affects nitrite accumulation rates. This accumulation is low, 0.05 to 0.10 g N-NO2(-) per g N-NO3(-) eliminated, for alcohols such as methanol, ethanol, or glycerol. The utilization of glycerol leads to fungal development causing clogging of the biofilters. This fungal growth and consequent clogging exclude this carbon source, with little nitrite accumulation, as carbon source for denitrification. Whatever the carbon source, the C/N ratio in the biofilter plays a major role in the appearance of residual nitrite; an optimal C/N ratio from 3.0 to 3.2 allows a complete denitrification without any nitrite accumulation.

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria.

An extracellular chitinase (ChiA-65) was produced and purified from a newly isolated Bacillus licheniformis LHH100. Pure protein was obtained after heat treatment and ammonium sulphate precipitation followed by Sephacryl S-200 chromatography. Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 65,195.13 Da. The sequence of the 27 N-terminal residues of the mature ChiA-65 showed high homology with family-18 chitinases. Optimal activity was achieved at pH 4 and 75 °C. Among the inhibitors and metals tested, p-chloromercuribenzoic acid, N-ethylmaleimide, Hg(2+), and Hg(+) completely inhibited enzyme activity. Chitinase activity was high on colloidal chitin, glycol chitin, glycol chitosane, chitotriose, and chitooligosaccharide. Chitinase activity towards synthetic substrates in the order of p-NP-(GlcNAc)n (n = 2-4) was p-NP-(GlcNAc)2 > p-NP-(GlcNAc)4 > p-NP-(GlcNAc)3. Our results suggest that ChiA-65 preferentially hydrolyzed the second glycosidic link from the non-reducing end of (GlcNAc)n. ChiA-65 obeyed Michaelis-Menten kinetics, the Km and kcat values being 0.385 mg, colloidal chitin/ml and 5000 s(-1), respectively. The chiA-65 gene encoding ChiA-65 was cloned in Escherichia coli and its sequence was determined. Above all, ChiA-65 exhibited remarkable biochemical properties suggesting that this enzyme is suitable for bioconversion of chitin waste.

Modeling of an aerobic bioprocess based on gas exchange and dynamics: a novel approach.

Monitoring of the biological degradation of a substrate by microorganisms is a key issue, especially in the soil bioremediation area. Respiration measurement is the easiest way to obtain online information on the biological activity. Nevertheless, it is indirectly related to substrate consumption and microbial growth. To be able to link these phenomena, a robust and descriptive model has been developed. Both biological and gas/liquid transfer dynamics must be taken into account to link the online measurement with the actual biological respiration. For that, experimental evolution of the respiratory ouotient (RQ) during a biodegradation has been compared against general biodegradation knowledge. To obtain a reliable model, practical and structural sensitivity analyses have been conducted. The model can describe the evolution of both online measurable and non-measurable states. It also gives a new definition of the apparent RQ, measured in the gas phase, compared to the actual biological RQ.

A simple and rapid one-time method to evaluate the non-acidic gas content from bioprocesses.

This paper presents a rapid less than 2 min and low-cost method involving the use of alkali solution to capture the acidic gasses from a biogas, thereby providing an estimate of the percentage of non-acidic gasses. Such a method was mentioned in the literature but never fully described or optimized. After sampling an aliquot of gas from bioprocess, gas was injected in a sealed flask with a 3 M NaOH solution, and after equilibrium was obtained, the non-acidic gas volume was measured. The method was first calibrated with certified gasses with an accuracy observed between 98 and 105%. Regarding the validation step, certified standard gas mixtures and nine biogas-laboratory batch reactors were used, the overall accuracy reported was 103 + 3%. This rapid and low-cost method may either be used in laboratory conditions as a quick and low cost alternative to standard analysis equipment or in addition as a routine field control method used on full-scale plants.

Municipal wastewater treatment by biofiltration: comparisons of various treatment layouts. Part 1: assessment of carbon and nitrogen removal.

One of the largest wastewater treatment plants in the Paris conurbation (240,000 m(3)/d) has been studied over several years in order to provide technical and economical information about biological treatment by biofiltration. Biofiltration systems are processes in which carbon and nitrogen pollution of wastewater are treated by ascendant flow through immersed fixed cultures. This paper, focused on technical information, aims: (1) to compare performances of the three biological treatment layouts currently used in biofiltration systems: upstream denitrification (UD), downstream denitrification (DD) and combined upstream-downstream denitrification (U-DD) layouts; and (2) to describe in detail each treatment step. Our study has shown that more than 90% of the carbon and ammoniacal pollution is removed during biological treatment, whatever the layout used. Nitrate, produced during nitrification, is then reduced to atmospheric nitrogen. This reduction is more extensive when the denitrification stage occurs downstream from the treatment (DD layout with methanol addition), whereas it is only partial when it is inserted upstream from the treatment (UD layout - use of endogenous carbonaceous substrate). So, the UD layout leads to a nitrate concentration that exceeds the regulatory threshold in the effluent, and the treatment must be supplemented with a post-denitrification step (U-DD layout). Our work has also shown that the optimal ammonium-loading rate is about 1.1-1.2 kg N-NH(4)(+) per m(3) media (polystyrene) and day. For denitrification, the optimal nitrate-loading rate is about 2.5 kg N per m(3) media (expanded clay) and day in the case of DD with methanol, and is about 0.25 kg N-NO(3)(-) per m(3) media and day in the case of UD with exogenous carbonaceous substrate.

Municipal wastewater treatment by biofiltration: comparisons of various treatment layouts. Part 2: assessment of the operating costs in optimal conditions.

This work aims to compare the operation costs (energy, reagents, waste management) for the three layouts usually used in wastewater treatment plants incorporating biofilters, using technical and economical data acquired during 10 years of operation of a Parisian plant (Seine Centre, 240,000 m(3) d(-1) -800,000 equivalent inhabitants). The final objective is to establish general economical data and tendencies that can be translated from our study to any biofiltration plant. Our results evidenced the savings achieved through the treatment process combining upstream and downstream denitrification. To use this layout reduced the operating costs by some 10% as compared with conventional processing only comprising downstream denitrification. Operating costs were respectively estimated at 37 and 34 €/1,000 m(3) for downstream denitrification and combining upstream and downstream denitrification layouts.

Antagonistic activity of Bacillus sp. obtained from an Algerian oilfield and chemical biocide THPS against sulfate-reducing bacteria consortium inducing corrosion in the oil industry.

The present study enlightens the role of the antagonistic potential of nonpathogenic strain B21 against sulfate-reducing bacteria (SRB) consortium. The inhibitor effects of strain B21 were compared with those of the chemical biocide tetrakishydroxymethylphosphonium sulfate (THPS), generally used in the petroleum industry. The biological inhibitor exhibited much better and effective performance. Growth of SRB in coculture with bacteria strain B21 antagonist exhibited decline in SRB growth, reduction in production of sulfides, with consumption of sulfate. The observed effect seems more important in comparison with the effect caused by the tested biocide (THPS). Strain B21, a dominant facultative aerobic species, has salt growth requirement always above 5% (w/v) salts with optimal concentration of 10-15%. Phylogenetic analysis based on partial 16S rRNA gene sequences showed that strain B21 is a member of the genus Bacillus, being most closely related to Bacillus qingdaonensis DQ115802 (94.0% sequence similarity), Bacillus aidingensis DQ504377 (94.0%), and Bacillus salarius AY667494 (92.2%). Comparative analysis of partial 16S rRNA gene sequence data plus physiological, biochemical, and phenotypic features of the novel isolate and related species of Bacillus indicated that strain B21 may represent a novel species within the genus Bacillus, named Bacillus sp. (EMBL, FR671419). The results of this study indicate the application potential of Bacillus strain B21 as a biocontrol agent to fight corrosion in the oil industry.

A new quantitative and low-cost determination method of nitrate in vegetables, based on deconvolution of UV spectra.

A new UV-spectrophotometric method for the determination of nitrate in vegetables is presented. The method is based on the spectral deconvolution: UV spectrum of a sample is considered as a linear combination of absorption spectra, named reference spectra. The combination of a small number of spectra of reference allows to reconstitute the shape of UV spectrum of an unknown sample. There have been several fresh vegetables (lettuce, curly lettuce, oak-leaf lettuce), as well as frozen spinaches that have been tested. The results obtained were comparable to those obtained with the reference HPLC method (official European reference method for the determination of nitrate in foodstuffs). The nitrate content varied from 377 to 3240 mg kg(-1) of fresh vegetables, and 545 to 1190 mg kg(-1) of frozen spinach. The recovery of added nitrate ranged from 91 to 99%. The results were obtained with a laboratory spectrophotometer and also with a dedicated field-type spectrophotometer. This method does not require almost any consumable, is quantitative and very fast reading with easy and low maintenance.

Aerobic degradation of ethyl-tert-butyl ether by a microbial consortium: selection and evaluation of biodegradation ability.

A microbial consortium that degrades ethyl-tert-butyl ether (ETBE) as the sole source of carbon and energy under aerobic conditions was selected from a gasoline-polluted soil. This consortium consists of a variety of microorganisms with a predominance of filamentous morphology. Degradation of ETBE was found to be solely related to bacterial activity. After prolonged cultivation followed by successive transfers, the consortium's degradation ability was improved and reached a specific degradation rate of 95 mg/g(protein)/h (about 146 mg/g(dry wt)/h). This exceeds the previously reported rates in the literature for ETBE-degrading microorganisms as pure or mixed cultures. Furthermore, a stoichiometric balance of chemical oxygen demand (COD) removal and oxygen uptake with ETBE removal provides indirect evidence of complete degradation. The consortium's activity was not inhibited by high ETBE concentrations (< or = 1,600 mg/L), and large inoculum sizes (> or = 120 mg(protein)/L) were desirable for a faster and complete degradation of ETBE. The enriched consortium was also able to completely degrade methyl-tert-butyl ether (MTBE), tert-amyl methyl ether (TAME), and tert-butyl alcohol (TBA). both alone and in mixture with ETBE, without any measurable release of major degradation intermediates. In each case, MTBE and TAME exhibited the most significant resistance to degradation while TBA was rapidly degraded.