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During in situ biomethanation, microbial communities can convert complex Organic Matter (OM) and H 2 into CH 4 . OM biodegradation was compared between Anaerobic Digestion (AD) and in situ biomethanation, in semi-continuous processes, using two inocula from the digester (D) and the post-digester (PoD) of an AD plant. The impact of H 2 on OM degradation was assessed using a fractionation method. Operational parameters included 20 days of hydraulic retention time and 1.5 g VS .L -1 .d -1 of organic loading rate. During in situ biomethanation, 485 NmL of H 2 were injected for each feeding (3 times a week). Maximum organic COD removal was 0.6 gCOD in AD control and at least 1.6 gCOD for in situ biomethanation. Therefore, COD removal was 2.5 times higher with H 2 injections. These results bring out the potential of H 2 injections during AD, not only for CO 2 consumption but also for better OM degradation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The performance of hydrogen consumption by various inocula derived from mesophilic anaerobic digestion plants was evaluated under ex situ biomethanation. A panel of 11 mesophilic inocula was operated at a concentration of 15 g VS .L -1 at a temperature of 35 °C in batch system with two successive injections of H 2 :CO 2 (4:1 mol:mol). Hydrogen consumption and methane production rates were monitored from 44 h to 72 h. Hydrogen consumption kinetics varies significantly based on the inoculum origin, with no accumulation of volatile fatty acids. Microbial community analyses revealed that microbial indicators such as the increase in Methanosarcina sp. abundance and the increase of the Archaea/Bacteria ratio were associated to high initial hydrogen consumption rates. The improvement in the hydrogen consumption rate between the two injections was correlated with the enrichment in hydrogenotrophic methanogens. This work provides new insights into the early response of microbial communities to hydrogen injection and on the microbial structures that may favor their adaptation to the biomethanation process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The conversion of H 2 into methane can be carried out by microorganisms in a process so-called biomethanation. In ex-situ biomethanation H 2 and CO 2 gas are exogenous to the system. One of the main limitations of the biomethanation process is the low gas-liquid transfer rate and solubility of H 2 which are strongly influenced by the temperature. Hydrogenotrophic methanogens that are responsible for the biomethanation reaction are also very sensitive to temperature variations. The aim of this work was to evaluate the impact of temperature on batch biomethanation process in mixed culture. The performances of mesophilic and thermophilic inocula were assessed at 4 temperatures (24, 35, 55 and 65 °C). A negative impact of the low temperature (24 °C) was observed on microbial kinetics. Although methane production rate was higher at 55 and 65 °C (respectively 290 ± 55 and 309 ± 109 mL CH 4 /L.day for the mesophilic inoculum) than at 24 and 35 °C (respectively 156 ± 41 and 253 ± 51 mL CH 4 /L.day), the instability of the system substantially increased, likely because of a strong dominance of only Methanothermobacter species. Considering the maximal methane production rates and their stability all along the experiments, an optimal temperature range of 35 °C or 55 °C is recommended to operate ex-situ biomethanation process.

Background: Biomethanation is a promising solution to upgrade the CH 4 content in biogas. This process consists in the injection of H 2 into an anaerobic digester, using the capacity of indigenous hydrogenotrophic methanogens for converting the injected H 2 and the CO 2 generated from the anaerobic digestion process into CH 4 . However, the injection of H 2 could cause process disturbances by impacting the microbial communities of the anaerobic digester. Better understanding on how the indigenous microbial community can adapt to high H 2 partial pressures is therefore required., Results: Seven microbial inocula issued from industrial bioprocesses treating different types of waste were exposed to a high H 2 partial pressure in semi-continuous reactors. After 12 days of operation, even though both CH 4 and volatile fatty acids (VFA) were produced as end products, one of them was the main product. Acetate was the most abundant VFA, representing up to 94% of the total VFA production. VFA accumulation strongly anti-correlated with CH 4 production according to the source of inoculum. Three clusters of inocula were distinguished: (1) inocula leading to CH 4 production, (2) inocula leading to the production of methane and VFA in a low proportion, and (3) inocula leading to the accumulation of mostly VFA, mainly acetate. Interestingly, VFA accumulation was highly correlated to a low proportion of archaea in the inocula, a higher amount of homoacetogens than hydrogenotrophic methanogens and, the absence or the very low abundance in members from the Methanosarcinales order. The best methanogenic performances were obtained when hydrogenotrophic methanogens and Methanosarcina sp. co-dominated all along the operation., Conclusions: New insights on the microbial community response to high H 2 partial pressure are provided in this work. H 2 injection in semi-continuous reactors showed a significant impact on microbial communities and their associated metabolic patterns. Hydrogenotrophic methanogens, Methanobacterium sp. or Methanoculleus sp. were highly selected in the reactors, but the presence of co-dominant Methanosarcinales related species were required to produce higher amounts of CH 4 than VFA., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2020.)

The aim of this article is to provide an analysis of microwave effects on ferulic and coumaric acids (FA and CA, respectively) extraction from grass biomass (corn stalks and miscanthus). Microwave pretreatment using various solvents was first compared to conventional heating on corn stalks. Then, microwave operational conditions were extended in terms of incident power and treatment duration. Optimal conditions were chosen to increase p -hydroxycinnamic acids release. Finally, these optimal conditions determined on corn stalks were tested on miscanthus stalks to underlie the substrate incidence on p -hydroxycinnamic acids release yields. The optimal conditions-a treatment duration of 405 s under 1000 W-allowed extracting 1.38% FA and 1.97% CA in corn stalks and 0.58% FA and 3.89% CA in miscanthus stalks. The different bioaccessibility of these two molecules can explain the higher or lower yields between corn and miscanthus stalks., Competing Interests: The authors declare no conflict of interest.

The effect of supplementing granular activated carbon and trace elements on the anaerobic digestion performance of consecutive batch reactors treating food waste was investigated. The results from the first batch suggest that addition of activated carbon favored biomass acclimation, improving acetic acid consumption and enhancing methane production. Adding trace elements allowed a faster consumption of propionic acid. A second batch proved that a synergy existed when activated carbon and trace elements were supplemented simultaneously. The degradation kinetics of propionate oxidation were particularly improved, reducing significantly the batch duration and improving the average methane productivities. Addition of activated carbon favored the growth of archaea and syntrophic bacteria, suggesting that interactions between these microorganisms were enhanced. Interestingly, microbial analyses showed that hydrogenotrophic methanogens were predominant. This study shows for the first time that addition of granular activated carbon and trace elements may be a feasible solution to stabilize food waste anaerobic digestion., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Anaerobic digestion of food waste is a complex process often hindered by high concentrations of volatile fatty acids and ammonia. Methanogenic archaea are more sensitive to these inhibitors than bacteria and thus the structure of their community is critical to avoid reactor acidification. In this study, the performances of three different inocula were compared using batch digestion tests of food waste and cardboard mixtures. Particular attention was paid to the archaeal communities in the inocula and after digestion. While the tests started with inocula rich in Methanosarcina led to efficient methane production, VFAs accumulated in the reactors where inocula initially were poor in this archaea and no methane was produced. In addition, higher substrate loads were tolerated when greater proportions of Methanosarcina were initially present in the inoculum. Independently of the inoculum origin, Methanosarcina were the dominant methanogens in the digestates from the experiments that efficiently produced methane. These results suggest that the initial archaeal composition of the inoculum is crucial during reactor start-up to achieve stable anaerobic digestion at high concentrations of ammonia and organic acids., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study evaluated the influence of the co-substrate proportions (0-60% of cardboard in dry basis) and the initial total solid contents (20-40%) on the batch fermentation performance. Maximum hydrogen yields were obtained when mono-fermenting food waste at high solids contents (89mlH 2 ·gVS -1 ). The hydrogen yields were lower when increasing the proportions of cardboard. The lower hydrogen yields at higher proportions of cardboard were translated into higher yields of caproic acid (up to 70.1gCOD·kgCOD bio -1 ), produced by consumption of acetic acid and hydrogen. The highest substrate conversions were achieved at low proportions of cardboard, indicating a stabilization effect due to higher buffering capacities in co-fermentation. Clostridiales were predominant in all operational conditions. This study opens up new possibilities for using the cardboard proportions for controlling the production of high added-value products in dry co-fermentation of food waste., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The objective of this study was to test three different alternatives to mitigate the destabilizing effect of accumulation of ammonia and volatile fatty acids during food waste anaerobic digestion. The three options tested (low temperature, co-digestion with paper waste and trace elements addition) were compared using consecutive batch reactors. Although methane was produced efficiently (∼500ml CH 4 gVS -1 ; 16l CH 4 lreactor -1 ), the concentrations of propionic acid increased gradually (up to 21.6gl -1 ). This caused lag phases in the methane production and eventually led to acidification at high substrate loads. The addition of trace elements improved the kinetics and allowed higher substrate loads, but could not avoid propionate accumulation. Here, it is shown for the first time that addition of activated carbon, trace elements and dilution can favor propionic acid consumption after its accumulation. These promising options should be optimized to prevent propionate accumulation., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Dry anaerobic digestion is a promising option for food waste treatment and valorization. However, accumulation of ammonia and volatile fatty acids often occurs, leading to inefficient processes and digestion failure. Co-digestion with cardboard may be a solution to overcome this problem. The effect of the initial substrate to inoculum ratio (0.25 to 1gVS·gVS -1 ) and the initial total solids contents (20-30%) on the kinetics and performance of dry food waste mono-digestion and co-digestion with cardboard was investigated in batch tests. All the conditions produced methane efficiently (71-93% of the biochemical methane potential). However, due to lack of methanogenic activity, volatile fatty acids accumulated at the beginning of the digestion and lag phases in the methane production were observed. At increasing substrate to inoculum ratios, the initial acid accumulation was more pronounced and lower cumulative methane yields were obtained. Higher amounts of soluble organic matter remained undegraded at higher substrate loads. Although causing slightly longer lag phases, high initial total solids contents did not jeopardize the methane yields. Cardboard addition reduced acid accumulation and the decline in the yields at increasing substrate loads. However, cardboard addition also caused higher concentrations of propionic acid, which appeared as the most last acid to be degraded. Nevertheless, dry co-digestion of food waste and cardboard in urban areas is demonstrated asan interesting feasible valorization option., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The increasing food waste production calls for developing efficient technologies for its treatment. Anaerobic processes provide an effective waste valorization. The influence of the initial substrate load on the performance of batch dry anaerobic co-digestion reactors treating food waste and cardboard was investigated. The load was varied by modifying the substrate to inoculum ratio (S/X), the total solids content and the co-digestion proportions. The results showed that the S/X was a crucial parameter. Within the tested values (0.25, 1 and 4gVS·gVS -1 ), only the reactors working at 0.25 produced methane. Methanosarcina was the main archaea, indicating its importance for efficient methanogenesis. Acidogenic fermentation was predominant at higher S/X, producing hydrogen and other metabolites. Higher substrate conversions (≤48%) and hydrogen yields (≤62mL·gVS -1 ) were achieved at low loads. This study suggests that different value-added compounds can be produced in dry conditions, with the initial substrate load as easy-to-control operational parameter., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, the effect of total solid content (TS) on thermophilic hydrogen production from wheat straw was investigated. Six TS contents ranging from wet to dry conditions (10-34%TS) were tested in batch tests. A decrease of H₂ yields was observed and three statistical groups were distinguished according to the TS content: wet conditions (10% and 14%TS) with 15.3 ± 1.6 NmlH₂ gTS(-1), intermediate conditions (19%TS) with 6.4 ± 1.0 NmlH₂ gTS(-1) and dry conditions (25-34%TS) with 3.4 ± 0.8 NmlH₂ gTS(-1). Such a decrease in biohydrogen yields was related to a metabolic shift with an accumulation of lactic acid under dry conditions. Concomitantly, a microbial population shift was observed with a dominance of species related to the class Clostridia under wet conditions, and a co-dominance of members of Bacilli, Clostridia classes and Bacteroidetes phylum under dry conditions., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, changes in bacterial and archaeal communities involved in anaerobic digestion processes operated with high solid contents were investigated. Batch tests were performed within a range of total solids (TS) of 10-35%. Between 10% and 25% TS, high methanogenic activity was observed and no overall specific structure of active bacterial communities was found. At 30% and 35%, methanogenesis was inhibited as a consequence of volatile fatty acids accumulation. Here, a specific bacterial signature was observed with three main dominant bacteria related to Clostridium sp., known for their ability to grow at low pH. Additionally, archaeal community was gradually impacted by TS content. Three archaeal community structures were observed with a gradual shift from Methanobacterium sp. to Methanosarcina sp., according to the TS content. Overall, several species were identified as biomarkers of methanogenesis inhibition, since bacterial and archaeal communities were highly specific at high TS contents., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The role of the total solids (TS) content on anaerobic digestion was investigated in batch reactors. A range of TS contents from 10% to 35% was evaluated, four replicates were performed. The total methane production slightly decreased with TS concentrations increasing from 10% to 25% TS. Two behaviors were observed at 30% TS: two replicates had similar performances to that at 25% TS; for the two other replicates, the methane production was inhibited as observed at 35% TS. This difference suggested that 30% TS content corresponded to a threshold of the solids content, above which methanogenesis was strongly inhibited. The Anaerobic Digestion Model No. 1 (ADM1) was used to describe the experimental data. The effects of hydrolysis step and liquid/gas mass transfer were particularly investigated. The simulations showed that mass transfer limitation could explain the low methane production at high TS, and that hydrolysis rate constants slightly decreased with increasing TS., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Anaerobic biofilm reactors have to be operated in a way that optimizes on one hand the start-up period by a quick growth of an active biofilm, on the other hand the regular operation by an active control of the biofilm to avoid diffusion limitations and clogging. This article is an overview of the research carried out at INRA-LBE for the last 15 years. The start-up of anaerobic biofilm reactors may be considerably shortened by applying a short inoculation period (i.e. contact between the inoculum and the support media). Then, the increase of the organic loading rate should be operated at a short hydraulic retention time and low hydrodynamic constraints in order to favor biofilm growth. After the start-up period, biofilm growth should be controlled to maintain a high specific activity and prevent clogging. This can be done in particulate biofilm systems by using hydrodynamics to increase or decrease shear forces and attrition but is much more difficult in anaerobic fixed bed reactors., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

This paper presents a review of the main sludge treatment techniques used as a pretreatment to anaerobic digestion. These processes include biological (largely thermal phased anaerobic), thermal hydrolysis, mechanical (such as ultrasound, high pressure and lysis), chemical with oxidation (mainly ozonation), and alkali treatments. The first three are the most widespread. Emphasis is put on their impact on the resulting sludge properties, on the potential biogas (renewable energy) production and on their application at industrial scale. Thermal biological provides a moderate performance increase over mesophilic digestion, with moderate energetic input. Mechanical treatment methods are comparable, and provide moderate performance improvements with moderate electrical input. Thermal hydrolysis provides substantial performance increases, with a substantial consumption of thermal energy. It is likely that low impact pretreatment methods such as mechanical and thermal phased improve speed of degradation, while high impact methods such as thermal hydrolysis or oxidation improve both speed and extent of degradation. While increased nutrient release can be a substantial cost in enhanced sludge destruction, it also offers opportunities to recover nutrients from a concentrated water stream as mineral fertiliser., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Estrogens, which contribute greatly to the endocrine-disrupting activity in sewage, are partially sorbed onto particulate matter during sewage treatment. We thus investigated the occurrence of estrogens in different kinds of sludge and throughout a plant-scale anaerobic digestion process. The analytical method was first validated when sorption interaction between spiked estrogens and sludge could occur. Hence, the recovery ratio of estrone (E1), 17beta-estradiol (E2), estriol (E3) and 17alpha-ethinylestradiol (EE2) were determined when added to liquid sludge and mixed under various conditions. We show that minor non-extractable residues were formed (5-10%), suggesting that the sorption interaction established with sludge did not limit estrogen extraction. Estrogen concentrations measured in collected samples varied with sludge type. Secondary sludge showed higher E1 contents than primary sludge: respectively, 43 and 8 ng g(-1) dry weight (dw). Two pathways of E1 production during secondary treatment are proposed to explain such a result. Higher estrogen concentrations were found in secondary sludge from a conventional plant (55 ng g(-1)dw) compared to those from an advanced plant (13 ng g(-1)dw). Based on estimated estrogen concentrations in sewage, we conclude that operating parameters play a role in the sorption of estrogens during secondary treatment. Also, the hydrophobic properties of the estrogens influenced the individual adsorption of each molecule. Thus, E3 showed the highest estimated concentrations in sewage but very low concentrations in sludge. Finally, plant-scale anaerobic digestion showed low efficiency (<40%) for removing estrogens and, regarding the final dewatering process, concentrations increased for E2 and EE2., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

A thermochemical pretreatment, i.e. saponification, was optimised in order to improve anaerobic biodegradation of slaughterhouse wastes such as aeroflotation grease and flesh fats from cattle carcass. Anaerobic digestion of raw wastes, as well as of wastes saponified at different temperatures (60 degrees C, 120 degrees C and 150 degrees C) was conducted in fed-batch reactors under mesophilic condition and the effect of different saponification temperatures on anaerobic biodegradation and on the long-chain fatty acids (LCFAs) relative composition was assessed. Even after increasing loads over a long period of time, raw fatty wastes were biodegraded slowly and the biogas potentials were lower than those of theoretical estimations. In contrast, pretreated wastes exhibited improved batch biodegradation, indicating a better initial bio-availability, particularly obvious for carcass wastes. However, LCFA relative composition was not significantly altered by the pretreatment. Consequently, the enhanced biodegradation should be attributed to an increased initial bio-availability of fatty wastes without any modification of their long chain structure which remained slowly biodegradable. Finally, saponification at 120 degrees C achieved best performances during anaerobic digestion of slaughterhouse wastes., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The biodegradation of estradiol (E2), estrone (E1), and ethinylestradiol (EE2) was investigated using mixed bacterial cultures enriched from activated sludge. Enrichments were carried out on E2 or EE2 in batch conditions with acetonitrile as additional carbon source. Degradation experiments were performed both using hormones as sole carbon source or with an additional source. The hormones were completely degraded by these cultures. Estradiol was rapidly converted to E1 within 24 h. Thereafter, E1 degradation began, displaying a lag phase ranging from 3 to 4 days. Estrone depletion took from 48 h to more than 6 days, depending on the culture conditions. For EE2 degradation, when it was the sole carbon source, the lag phase and the time required for its complete removal (7 and 15 days, respectively) were shorter that in cultures with a supplementary carbon source. The specific degradation rates observed for E2 both with and without an additional carbon source were similar. By contrast, the specific degradation rates for E1 and EE2 were, respectively, seven and 20 times faster when these hormones were supplied as the sole carbon source. The bacterial community structure of each culture was characterized by molecular and cultural methods. The mixed cultures were made up of species belonging to Alcaligenes faecalis, Pusillimonas sp., Denitrobacter sp., and Brevundimonas diminuta or related to uncultured Bacteroidetes. The isolated strain B. diminuta achieved the conversion of E2 to E1.

The thermochemical pretreatment by saponification of two kinds of fatty slaughterhouse waste--aeroflotation fats and flesh fats from animal carcasses--was studied in order to improve the waste's anaerobic degradation. The effect of an easily biodegradable compound, ethanol, on raw waste biodegradation was also examined. The aims of the study were to enhance the methanisation of fatty waste and also to show a link between biodegradability and bio-availability. The anaerobic digestion of raw waste, saponified waste and waste with a co-substrate was carried out in batch mode under mesophilic and thermophilic conditions. The results showed little increase in the total volume of biogas, indicating a good biodegradability of the raw wastes. Mean biogas volume reached 1200 mL/g VS which represented more than 90% of the maximal theoretical biogas potential. Raw fatty wastes were slowly biodegraded whereas pretreated wastes showed improved initial reaction kinetics, indicating a better initial bio-availability, particularly for mesophilic runs. The effects observed for raw wastes with ethanol as co-substrate depended on the process temperature: in mesophilic conditions, an initial improvement was observed whereas in thermophilic conditions a significant decrease in biodegradability was observed.

PAH are particularly monitored because of their carcinogenic properties and their ubiquity in the environment. Their presence in municipal sewage sludge is a major problem due to the environmental risks associated with the sludge spreading on agricultural soils. The objective of this work was to asses the removal of PAH naturally present in sludge by continuous anaerobic digestion with recirculation of ozonated sludge. Recirculation of ozonated digested sludge allowed to enhance PAH removals, the highest efficiency was obtained with the highest ozone dose (0.11gO(3)/g(TS)). In order to study the effect of recirculation, a reactor was operated without recirculation but was fed with a mixture of raw and ozonated digested sludge. This process led to the best performances in terms of PAH and solid removals. This pointed out some accumulation of nonbiodegradable or recalcitrant compounds during recirculation assay. Smallest and most soluble compounds presented the highest biodegradation efficiencies.

Bioaugmentation experiments with the aerobic denitrifier Microvirgula aerodenitrificans were performed in an aerobic continuous stirred tank reactor (CSTR) treating urban wastewater. The fate of the added bacteria was monitored by a specific fluorescent oligonucleotide probe targeting 16S rRNA. The first addition of the strain led to its rapid disappearance because of grazing. Bacteria were then embedded within an alginate matrix before inoculation. Alginate fragments adhered to the existing flocs and were progressively colonized by the indigenous flora. Thereafter, microcolonies of the exogenous bacterium were found to be incorporated into existing flocs.

An anaerobic sequencing batch reactor (AnSBR) was used to treat the dissolved air flotation skimmings from a cooked pork meat plant. During the start-up period, the reactor was operated in fed-batch mode for 25 days and 7 batches were treated. The SBR was inoculated with sludge taken from a reactor treating distillery vinasse. The results showed that this kind of sludge is a very good source of inoculum for digesters treating residues with a high content in fats and long-chain fatty acids because it was able to adapt very rapidly to the new substrate and, from the second batch on, the sludge was already able to metabolize the fatty residue at quite high rates. The AnSBR was then operated with 5 batches per week for 110 days and the quantity of VS added per batch was regularly increased until the maximum treatment capacity of the reactor (i.e. maximum loading rate) was reached. The maximum organic loading rates were found to be 0.16 g VS/g VSS d, or 0.224 g VS/g VSS.batch when the reactor is fed 5 times a week. The biodegradability of the skimmings was very high, with more than 97% of TS removal, and the methane production was 880+/-90 mL of methane/g of VS(added).

Thermal treatments with temperature ranging from 60 to 210 degrees C were applied to 6 waste-activated sludge samples originating from high or medium load, extended aeration wastewater treatment processes that treated different wastewaters (urban, urban and industrial or slaughterhouse). COD sludge solubilisation was linearly correlated with the treatment temperature on the whole temperature range and independently of the sludge samples. Sludge batch mesophilic biodegradability increased with treatment temperature up to 190 degrees C. In this temperature range, biodegradability enhancement or methane production increase by thermal hydrolysis was shown to be a function of sludge COD solubilisation but also of sludge initial biodegradability. The lower the initial biodegradability means the higher efficiency of thermal treatment.

The steroid hormones estrone (E(1)), 17beta-estradiol (E(2)), estriol (E(3)), 17alpha-ethinylestradiol (EE(2)), and their conjugated forms were surveyed throughout an advanced sewage treatment plant (STP). The estrogen concentrations in water and sludge samples, collected in October 2004 and April 2005, were determined by gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry. Simultaneously, the estrogenic activity was quantified using estrogen-responsive reporter cell lines (MELN) to investigate the behavior of overall estrogenic compounds. The estrogen concentrations in the inlet ranged from 200 to 500 ng/L, with the contribution of conjugated forms being higher than 50%. The major estrogens in influent were E(1) and E(3). The estrogenic activity was between 25 and 130 ng/L of E(2) equivalents (EEQs). Estrogen concentrations and estrogenicity measured in the inlet and in primary treated sewage were similar, showing a weak impact of primary treatment on hormone removal. In contrast, both estrogen concentration and estrogenicity decreased during biological treatment, with high removal efficiencies (>90%). Estrone, E(2), and EE(2) persisted in the treated water below 10 ng/L, whereas the estrogenicity was lower than 5 ng/L of EEQs. Estrogen mass flux in the effluent and sludge represented less than 2 and 4%, respectively, of the inlet. Consequently, the fraction of estrogens sorbed into the sludge was very small, and biodegradation was the main vehicle for estrogen elimination. This dual approach, comparing chemical and biological analysis, allowed us to confirm that most of the estrogenic activity occurring in this STP, which receives mainly domestic sewage, resulted from sex hormones.

The influence of the hydraulic retention time (HRT) on the start-up phase of a methanogenic inverse turbulent bed bioreactor was investigated. Two identical reactors were monitored, the only differing parameter being the HRT: one of the reactors was fed with a diluted wastewater at a constant HRT of 1 day, the organic loading rate (OLR) being increased by decreasing the substrate dilution; the second reactor was fed at a constant influent concentration of 20 g COD L(-1), the OLR being increased by decreasing the HRT from 40 days to 1 day. After 45 days of start-up, both reactors were operated at an OLR of 20 g COD L(-1)d(-1) and a HRT of 1 day. However, strong differences were observed on biofilm growth. In the reactor operated at a constant short HRT, biofilm concentration was 4.5 as high as in the reactor operated at an increasing HRT. This difference was attributed to the competition between planktonic and biofilm microorganisms in the latter reactor, whereas suspended biomass was quickly washed out in the former reactor because of the low HRT.

The purpose of this work was to study the anaerobic digestion of by-products generated during the production and refining of oil with the objective of proposing an alternative solution (methanisation) to the conventional solutions while reducing the energy consumption of fossil origin on refinery sites. The production of sunflower oil was taken as example. Glycerine from the production of biodiesel was also included in this study. The results show that glycerine has a high potential for methanisation because of its high methane potential (465 ml CH4/g VS) and high metabolization rates (0.42 g VS/g VSS.d). The use of oil cake as substrate for anaerobic digestion is not interesting because it has a low methane potential of 215 ml CH4/g VS only and because it is easily recovered in animal feed. Six residues have quite a high methane potential (465 to 850 ml CH4/g VS) indicating a good potential for anaerobic digestion. However, they contain a mixture of rapidly and slowly biodegradable organic matter and the loading rates must remain quite low (0.03 to 0.09 g VS/g VSS.d) to prevent any accumulation of slowly biodegradable solids in the digesters., (IWA Publishing 2008.)

The anaerobic digestion of the liquid residue (gelatinous water) coming from the production of fat from animal residue, was studied at laboratory and pilot scale. Biodegradability (>98%) and biogas potential (675 mL of biogas/g of COD(applied)) of this wastewater are very high. However, due to the high content on nitrogen, an inhibition of the anaerobic activity was observed for quite low concentrations of N-NH(3). Dilution of the wastewater and pH regulation in the reactor around 7.3 are the 2 solutions which were investigated to overcome the nitrogen inhibition at industrial scale. These two solutions were validated at laboratory scale in an anaerobic SBR and then onsite at pilot scale in a continuous reactor. A stable anaerobic digestion was observed in both reactors showing that no nitrogen inhibition was obtained when N-NH(3) concentration in the reactor was kept low.

Two enteric pathogens, Salmonella spp. and Campylobacter jejuni, and two bacteria commonly used as indicators, Escherichia coli and Clostridium perfringens, were monitored using quantitative real-time PCR during municipal wastewater treatment and sludge composting. The results were compared with those obtained using standard culture methods. A reduction of all bacteria was observed during wastewater treatment and during the thermophilic phase of composting. However, the bacterial groups studied behaved differently during the process, and the main differences were observed during biological treatment in activated sludge basins. In particular, Salmonella spp. and C. jejuni survived better during activated sludge treatment than E. coli. C. jejuni was the most resistant to wastewater treatment among the four bacterial groups. Overall, differences in survival were observed for all bacteria studied, when submitted to the same environmental pressure. This holds both for differences between indicators and pathogenic bacteria and between pathogenic bacteria. These results show the difficulty in defining reliable indicators.

A mathematical model based on Activated Sludge Model No. 3 (International Water Association, London) and laboratory-scale experiments were used to investigate ammonia conversion by nitrification in a sequencing batch reactor (SBR). The purpose of the study was to assess the effect of dissolved oxygen concentration on nitrite accumulation in the SBR. As the dissolved oxygen concentration in the SBR depends on the balance between oxygen consumption and oxygen transfer rates, ammonium conversion was measured for different air flowrate values to obtain different dissolved oxygen concentration profiles during the cycle. The ammonia concentration in the feeding medium was 500 mg ammonium as nitrogen (N-NH4(+))/L, and the maximum nitrite concentration achieved during a cycle was approximately 50 mg nitrite as nitrogen (N-NO2)/L. The air flow supplied to the reactor was identified as a suitable parameter to control nitrite accumulation in the SBR. This identification was carried out based on experimental results and simulation with a calibrated model. At a low value of the volumetric mass-transfer coefficient (kLa), the maximum nitrite concentration achieved during a cycle depends strongly on k(L)a, whereas, at a high value of k(L)a, the maximum nitrite concentration was practically independent of kL(a).

Polycyclic aromatic hydrocarbons are ubiquitous persistent pollutants. They may accumulate in sludge during wastewater treatment because of their low biodegradability and their hydrophobic characteristics. Combination of ozonation and anaerobic digestion may be efficient to remove PAHs naturally present in sludge. The objective of this study was to investigate the impact of ozone pre-treatment, with and without surfactant addition, on the anaerobic degradation of 12 PAHs (from low to high molecular weight). Under anaerobic digestion without ozonation pre-treatment, the highest removals were obtained for the lightest PAHs (3-aromatic rings). Ozonation pre-treatment of sludge allowed to increase biodegradability or bioavailability of each PAH, and the PAH removals were well correlated to the PAH solubility. Finally, addition of tyloxapol before sludge ozone pre-treatment had antagonist effects on PAH removal during anaerobic digestion: negative impact on anaerobic ecosystem activity and improvement of PAH bioaccessibility (particularly the PAHs with the highest octanol water partition coefficients).

The anaerobic treatment of saline effluents using halophilic and halotolerant microbial consortia is of major interest. Inhibition of anaerobic digestion is known to occur at high salt content. However, it seems that the suitable adaptation of an anaerobic sludge makes possible the treatment of saline wastewater. In this study, a non-saline anaerobic sludge was inoculated in two anaerobic batch reactors operating with a different substrate (distillery vinasse and ethanol) and subjected to increasing NaCl concentrations. The performance of the digesters appeared to be highly dependent on the nature of the substrate, and a similar level of inhibition (i.e. around 90% of the specific loading rate and specific methanogenic activity) was stated at 10 g l(-1) of NaCl with distillery vinasse and 60 g l(-1) of NaCl with ethanol. The characterization of the microflora and its adaptation to increasing NaCl conditions were also investigated using molecular tools based on the analysis of genomic 16S rDNA. The microbial communities revealed a high diversity that could be maintained in both reactors despite the increase in NaCl concentrations.

The influence of hydrodynamic conditions on the start-up phase of an inverse turbulent bed bioreactor was investigated. Two identical reactors, differing only by the gas velocity ensuring the carrier fluidization and generating the main hydrodynamic strengths (attrition), were monitored. Regarding the first 96 days, the experiments showed better performances for the reactor having the lowest hydrodynamic strengths. These results were correlated to lower attached biomass and higher biofilm specific removal rate for the reactor subjected to strong hydrodynamic strengths. Once the start-up was completed and the reactor stabilized at an organic loading rate of 6g(COD)L(-1)d(-1), the same hydrodynamic strengths were applied (gas velocity was equalized). The results proved that hydrodynamic conditions make it possible to control the biofilm characteristics. Consequently, it should be recommended to apply minimal strengths to enhance the biofilm growth during the early start-up phase, and then to increase them to control biofilm thickness and to optimize the reactor performances.

The microbial community of a pig slurry on a farm was monitored for 6 months using both molecular and cultural approaches. Sampling was carried out at all the different stages of effluent handling, from the rearing build-up to slurry spreading. Total DNA of each sample was extracted and analyzed by PCR-single-strand conformation polymorphism (SSCP) analysis using primers targeting the 16S rRNA genes from the archaeal and bacterial domains and also the Eubacterium-Clostridium, Bacillus-Streptococcus-Lactobacillus, and Bacteroides-Prevotella groups. A comparison of the SSCP profiles showed that there were rapid changes in the dominant bacterial community during the first 2 weeks of anaerobic storage and that the community was relatively stable thereafter. Several bacterial populations, identified as populations closely related to uncultured Clostridium and Porphyromonas and to Lactobacillus and Streptococcus cultured species commonly isolated from pig feces, remained present and dominant from the rearing build-up to the time of spreading. Enumeration of fecal indicators (enterococci and Escherichia coli) performed in parallel using cultural methods revealed the same trends. On the other hand, the archaeal community adapted slowly during pig slurry storage, and its diversity increased. A shift between two hydrogenotrophic methanogenic Methanobrevibacter populations from the storage pit to the pond was observed. Microorganisms present in pig slurry at the time of spreading could not be detected in soil after spreading by either molecular or cultural techniques, probably because of the detection limit inherent in the two techniques.

The exact extent of microbial diversity remains unknowable. Nevertheless, fingerprinting patterns [denaturing gradient electrophoresis (DGE), single-strand conformation polymorphism (SSCP)] provide an image of a microbial ecosystem and contain diversity data. We generated numerical simulation fingerprinting patterns based on three types of distribution (uniform, geometric and lognormal) with a range of units from 10 to 500,000. First, simulated patterns containing a diversity of around 1000 units or more gave patterns similar to those obtained in experiments. Second, the number of bands or peaks saturated quickly to about 35 and were unrelated to the degree of diversity. Finally, assuming lognormal distribution, we used an estimator of diversity on in silico and experimental fingerprinting patterns. Results on in silico patterns corresponded to the simulation inputs. Diversity results in experimental patterns were in the same range as those obtained from the same DNA sample in molecular inventories. Thus, fingerprinting patterns contain extractable data about diversity although not on the basis of a number of bands or peaks, as is generally assumed to be the case.

An anaerobic-anoxic sequencing batch reactor (A2 SBR) coupled with a fixed-bed nitrification reactor for simultaneous carbon, nitrogen and phosphorus removal was evaluated using slaughterhouse wastewater. Whereas the treatment could not be successfully carried out on the raw wastewater, the process showed very good nutrient removal performances after prefermentation. The removals of COD, N-NH4 and P-PO4 achieved were 99%, 85% and 99%, respectively. The increase in volatile fatty acid (VFA) and phosphate concentrations in the effluent after prefermentation may explain the high levels of biological carbon, nitrogen and phosphorus removal observed. A simple prefermentation is, therefore, necessary but sufficient to ensure good performances of the denitrifying enhanced biological phosphorus removal (EBPR) process.

The microbial community structure of pig manure slurry (PMS) was determined with comparative analysis of 202 bacterial, 44 archaeal and 33 eukaryotic small subunit (SSU) rDNA partial sequences. Based on a criterion of 97% of sequence similarity, the phylogenetic analyses revealed a total of 108, eight and five phylotypes for the Bacteria, Archaea and Eukarya lineages, respectively. Only 36% of the bacterial phylotypes were closely related (>or=97% similarity) to any previously known sequence in databases. The bacterial groups most often represented in terms of phylotype and clone abundance were the Eubacterium (22% of total sequences), the Clostridium (15% of sequences), the Bacillus-Lactobacillus-Streptococcus subdivision (20% of sequences), theMycoplasma and relatives (10% of sequences) and the Flexibacter-Cytophaga-Bacteroides (20% of sequences). The global microbial community structure and phylotype diversity show a close relationship to the pig gastrointestinal tract ecosystem whereas phylotypes from the Acholeplasma-Anaeroplasma and the Clostridium purinolyticum groups appear to be better represented in manure. Archaeal diversity was dominated by three phylotypes clustering with a group of uncultured microorganisms of unknown activity and only distantly related to the Thermoplasmales and relatives. Other Archaea were methanogenic H2/CO2 utilisers. No known acetoclastic Archaea methanogen was found. Eukaryotic diversity was represented by a pluricellular nematode, two Alveolata, a Blastocystis and an Entamoebidae. Manure slurry physico-chemical characteristics were analysed. Possible inhibitory effects of acetate, sulphide and ammonia concentrations on the microbial anaerobic ecosystem are discussed.

The acclimatisation of activated sludge to enhanced biological phosphorus removal (EBPR) conditions requires a period of about 40-100 days but its output remains hazardous. The impact of bioaugmentation on the start-up of a laboratory scale EBPR sequencing batch reactor was evaluated by process parameters measurement and microbial community dynamics monitoring using 16S rDNA targeted polymerase chain reaction-single strand conformation polymorphism electrophoresis (PCR-SSCP). Bioaugmentation: (1) speeded up the installation of good and stable EBPR in the bioaugmented reactor by about 15 days; (2) correlated with the transient enrichment of the sludge in the added microbial populations; and (3) favoured the long-term enrichment of the sludge in the phosphorus-accumulating organism (PAO) Candidatus Accumulibacter phosphatis. However, despite a lag time period, the control non-bioaugmented reactor ended up with comparable reactor parameters and microbial community evolution, suggesting that the same PAO populations were already present from the beginning in the original non-P-accumulating seed sludge. The potential of a true installation of the added microbial populations within the bioaugmented reactor compared to their substitution by indigenous similar populations is discussed. Competition between PAOs and the antagonistic glycogen accumulating organism Candidatus Competibacter phosphatis is also highlighted during EBPR start-up.

A recent PCR detection technique (TaqMan) based on a 3'-Minor Groove Binder (MGB) probe was applied to the detection of fecal-dominant bacteria to assess fecal contamination in environmental samples. Primers and probes used bacterial 16S ribosomal DNA (16S rDNA) as a gene marker and accurately defined with specificity a cluster of phylotypes within the Gram-positive low GC division. This cluster of phylotypes, called Fec1, corresponds to around 5% of human fecal microflora. Fec1 clustered 16S rDNA and strains (Eubacterium rectale) of fecal origin. A range of samples made up of feces and intestinal samples from humans and animals tested positive whereas other microbial ecosystems (soils, laboratory reactor, subsurface water) were negative. In order to circumvent problems related to DNA extraction efficiency, quantitative results took the form of the ratio between Fec1 16S rDNA and total bacterial 16S rDNA. The threshold of detection, defined as the ratio between Fec1 and total 16S rDNA, was measured as 0.006%.

The biological degradation of solid fatty residues is limited by their low bioavailability. In this work, the effect of the day mineral bentonite on the degradation of hexane extractable matter and its conversion to volatile fatty acids was investigated. Our results showed that the best performance in the elimination of hexane extractable matter (73% +/- 2.5) and the production of volatile fatty acids (39% +/- 1.5) were observed in continuous experimental assays with a bentonite/greases ratio of 0.9. Isotherm studies showed that the sorption equilibria of oleate on bentonite and sludge could be described by the LANGMUIR model.

Biological ammonium oxidation was carried out in two inverse turbulent bed reactors fed with synthetic mineral wastewater containing a high ammonium concentration (100 mg N-NH4+/L). Both reactors were started-up and operated in the same conditions except for the solid carrier concentration: the solid hold-up ratios applied, defined as the ratios of static to expanded bed height, were 0.1 and 0.3 in reactors R10 and R30 respectively. These two solid hold-up ratios generate different particle-to-particle collision frequencies and, therefore, detachment forces. The influence of solid hold-up on biofilm growth and nitrifying performance was studied from a macroscopic (i.e. nitrate and/or nitrite production) and microbiological point of view. After 60 days of operation, both reactors contained the same amount of biomass. However, R10 produced only nitrate while nitrite accumulated in R30. A comparison of microbial populations in the reactors showed that R10 contained both ammonium and nitrite oxidizing populations such as Nitrosomonas and Nitrospira, whereas in R30, ammonium oxidizing populations were much greater than those of nitrite oxidizers. The major ammonium-oxidizing organism was not the same in both reactors.

In France, fatty residues considered as "non-ultimate" waste will not be accepted in landfill from 2002, in accordance with French legislation. Anaerobic digestion appears as an alternative process to mobilize and profitably use such fermentable waste. In this work, the effect of an alkaline pretreatment on the degradation of hexane extractible matter (HEM) and the production of volatile fatty acids (VFAs) was compared in reactors working at pH 6.5 and 8.5. The results obtained showed that 40% (+/- 0.1) of HEM were degraded at pH 8.5 versus 10% (+/- 0.3) at pH 6.5, regardless of the alkali agent used to saponify the greasy wastes. The highest performances of VFA production (8.45% +/- 0.3) were obtained at pH 8.5 with greases saponified by potassium hydroxide, compared to results (4.25% +/- 0.1) with greasy waste saponified by sodium hydroxide. This difference in VFA production might be attributable to biotoxic molecules generated during the saponification of greases by soda, limiting consequently the VFA production process.

This paper presents a study about the influence of gas velocity on a methanogenic biofilm in an inverse turbulent bed reactor. Experimental results indicate a dynamic response of the growing attached biomass to the changes of hydrodynamic conditions, mainly attrition constraints. Short but intensive increases of gas velocity (U(g)) are shown to induce more detachment than a high but constant gas flow rate. Hydrodynamic conditions control the composition of the growing biofilm in terms of cells and exocellular polymeric substances (EPS). The cell fraction within the biofilm (R(cell)) was found to be inversely proportional to the gas velocity. The specific activity expressed in methane production rate or COD removal rate is higher in biofilms formed under high hydrodynamic constraints. The control of the hydrodynamic conditions in a biofilm reactor should make it possible to obtain a resistant and active biofilm.

The aim of the study was to determine the performances of a combined ozone/anaerobic digestion system for waste activated sludge reduction. The objective was the estimation of the process efficiency and stability when keeping constant influent flow while increasing recycled chemically treated flow. The ozonation step consisted in a partial oxidation (0.16 g O3/g SS) of the anaerobic mesophilic digested sludge. Chemical treatment of digested sludge resulted in a threefold COD solubilization and a decrease of SS of 22%. Some of the advantages of digested sludge ozonation were: deodorization, better settlement and a reduction in viscosity. However there were drawbacks: foaming during ozonation and, at high ozone doses, poorer filterability. The anaerobic digestion was carried out over 6 months with an increasing recycling of ozonated flow. Suspended solids removal rate and COD removal rate were compared with initial operating conditions for the biological reactor and the whole combined process. The optimum recycling rate was 25% with increases of SS removal and COD removal of 54% and 66% respectively when considering the combined process; corresponding to a decrease of the hydraulic retention time from 24 days to 19 days.

Treatment of winery wastewater was investigated using an anaerobic sequencing batch reactor (ASBR). Biogas production rate was monitored and permitted the automation of the bioreactor by a simple control system. The reactor was operated at an organic loading rate (ORL) around 8.6 gCOD/L.d with soluble chemical oxygen demand (COD) removal efficiency greater than 98%, hydraulic retention time (HRT) of 2.2 d and a specific organic loading rate (SOLR) of 0.96 gCOD/gVSS.d. The kinetics of COD and VFA removal were investigated for winery wastewater and for simple compounds such as ethanol, which is a major component of winery effluent, and acetate, which is the main volatile fatty acid (VFA) produced. The comparison of the profiles obtained with the 3 substrates shows that, overall, the acidification of the organic matter and the methanisation of the VFA follow zero order reactions, in the operating conditions of our study. The effect on the gas production rate resulted in two level periods separated by a sharp break when the acidification stage was finished and only the breaking down of the VFA continued.

The microbial community of a conventional anaerobic-aerobic sequencing batch reactor was investigated by cloning and sequencing bacterial 16S rDNA. The 92 16S rDNA sequences analysed ranged across 50 different operational taxonomic units (OTU). The majority of these sequences were not closely related to known species. They belonged to 12 different groups, but essentially to the Cytophagales and the Proteobacteria beta, which represented 38% and 17% of the retrieved sequences respectively. No OTU numerically outnumbered the others. However, similarities were observed with previous reports on molecular characterisation of phosphorus-accumulating ecosystems, suggesting an enrichment in microorganisms belonging to the Rhodocyclus group. Thereafter, the ability of this anaerobic-aerobic microbial community to accumulate phosphorus with nitrate as its energy source was investigated. The reactor was shifted from anaerobic-aerobic running conditions to anaerobic-anoxic conditions by injection of nitrate; and its microbial community was monitored by PCR-single strand conformation polymorphism (SSCP). The reactor maintained a good phosphorus accumulation and similar SSCP microbial community patterns for a period of 17 days, suggesting that the same microbial community was able to respire both oxygen and nitrate. However, this situation was unstable, since a breakdown in phosphorus accumulation occurred thereafter.

Optimizing anoxic biological phosphorus removal in the anaerobic-anoxic sequencing batch reactor (A2 SBR) was observed to depend on two parameters: the amount of added nitrate and the sludge retention time (SRT). The concentration of 120 mg N-NO3.l-1 in the anoxic medium and the SRT of 15 days were determined as optimal for a complete phosphorus removal in the A2 SBR. The reactor was supplied with synthetic wastewater containing 800 mg COD.l-1 acetic acid, 240 mg N-NH4.l-1 and 30 mg P-PO4.l-1. This study was completed by microscopic observations which revealed three morphological types of phosphate-accumulating bacteria (PAB).

Two laboratory-scale sequencing batch reactors were used to study biological treatment of carbon and nitrogen in a new combined anaerobic-aerobic system. Piggery wastewater was used as a model effluent. The anaerobic reactor, fed with raw wastewater and nitrite/nitrate recycling from the aerobic reactor, carried out denitrification and anaerobic digestion of organic carbon. In the aerobic reactor, organic carbon removal and nitrification of ammonia to mainly nitrite occurred. Three recycle-to-influent ratios from 1 to 3 were tested. The higher the recycle-to-influent ratio, the lower the concentrations of nitrogen oxides in the final effluent. Carbon removal efficiency was quite stable, whatever the ratio. However, the effect of this ratio was attenuated because of denitrification in the aerobic reactor, which increased the performances of the process. The use of sequencing batch reactors was essential to apply the configuration proposed, in order to combine denitrification and methanogenesis that require antagonistic conditions.

The acclimatisation of an activated sludge to enhanced biological phosphorus removal conditions was followed after and without bioaugmentation with a low amount of phosphorus-accumulating sludge. Phosphorus removal yields were monitored by conventional analytical methods and microbial communities evolutions were followed by a finger printing molecular technique (PCR-SSCP). While the benefit of the bioaugmentation seems real at the level of the reactor parameters, bioaugmentation speeded up the installation of good and stable phosphorus removal yield, the establishment of the inoculated microbial community in the bioaugmented reactor is still unclear. Both the bioaugmented and the control microbial communities evolved in a similar way to end up with apparently comparable populations. At the time of the experiment, the results suggest that the microbial community inoculated for the bioaugmentation did not establish in the reactor but compensated for phosphorus accumulation until the acclimatisation of an endogenous microbial community arose.

Four samples of natural ecosystems and one sample from an activated sludge treatment plant were mixed together and progressively adapted to alternating aerobic/anoxic phases in the presence of nitrate in order to enrich the microflora in aerobic denitrifiers. Aerobic denitrifying performances of this mixed ecosystem at various dissolved oxygen concentrations and various carbon-nitrogen loads were evaluated and compared to those obtained with the aerobic denitrifier Microvirgula aerodenitrificans. The consortium and the pure strain exhibited an aerobic denitrifying activity at air saturation conditions (7 mg dissolved oxygen 1(-1)), i.e. there was co-respiration of the two electron acceptors with significant specific nitrate reduction rates. Dissolved oxygen concentrations had no influence on denitrifying performances above a defined threshold: 0.35 mg l(-1) for the consortium and 4.5 mg l(-1) for M. aerodenitrificans respectively. Under these thresholds, decreasing the dissolved oxygen concentrations enhanced the denitrifying activity of each culture. The higher the carbon and nitrogen loads, the higher the performance of the aerobic denitrifying ecosystem. However, for M. aerodenitrificans, the nitrate reduction percentage was affected more by variations in nitrogen load than in carbon load.