References of "Thonart, Philippe"
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See detailPhysiological and bio-functional properties of gum arabic: a notable interest for certain human diseases
Eloundou Mballa, Pierre; Goffin, Dorothée ULg; Destain, Jacqueline ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (in press)

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See detailWood digestion in lower termites: multidisciplinary approaches based on differential feeding
Bauwens, Julien ULg; Brasseur, Catherine ULg; Tarayre, Cédric ULg et al

Poster (2014, December)

Termites digestive tract and hindgut especially still holds many secrets despites hundreds of years of research. The complexity of the symbiotic microbial community and the contrast of physio-chemical ... [more ▼]

Termites digestive tract and hindgut especially still holds many secrets despites hundreds of years of research. The complexity of the symbiotic microbial community and the contrast of physio-chemical environments found in lower termites paunch are potentially the key point to explain the efficiency of ligno-cellulose digestion. Contribution of advancing technologies accelerates the progress of our knowledge in this field. Here, we present multiple approaches combining old and recent techniques used to highlight the effect of ligno-cellulosic compounds on termite gut and the role of populations from the symbiotic microbial community. Termites Reticulitermes flavipes (Kollar) submitted to various artificial diets showed variations in flagellates populations profile and enzymatic activities. Differential protein expression was investigated using 2D-DIGE MALDI-TOF-TOF and 2D-LC-MS/MS using high resolution orbitrap analyzer. Results from both proteomic experiments tend to support each-other and bring complementary points of view. The gel-free analysis resulted in highly contrasted identification of enzymes involved in ligno-cellulose digestion and metabolism. Finally, differential feeding experiments leaded to in vivo selection of different symbiotic communities. These communities were characterized following some metabolism assays and allowed the cultivation of diverse microbial consortia using media closely related to the respective artificial diets. This work provides relevant data on termite and associated microbial community response to alimentary diets. [less ▲]

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See detailIsolation and cultivation of xylanolytic and cellulolytic Sarocladium kiliense and Trichoderma virens from the gut of the termite Reticulitermes santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

in Environmental Science & Pollution Research (2014)

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose ... [more ▼]

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose (with and without lignin) and beech wood xylan were used as diets instead of poplar wood in order to select cellulose and hemicellulose-degrading fungi. The strain Sarocladium kiliense (Acremonium kiliense) CTGxxyl was isolated from the termites fed on xylan, while the strain Trichoderma virens CTGxAviL was isolated from the termites fed on cellulose (with and without lignin). Both molds were cultivated in liquid media containing different substrates: agro-residues or purified polymers. S. kiliense produced maximal β-glucosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase and endo-1,4-β-D-xylanase activities of 0.103, 3.99, 0.53, and 40.8 IU/ml, respectively. T. virens produced maximal β-xylosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase, and endo-1,4-β-D-xylanase activities of 0.38, 1.48, 0.69, and 426 IU/ml. The cellulase and the xylanase of S. kiliense, less common than T. virens, were further investigated. The optimal activity of the xylanase was observed at pH 9–10 at 60 °C. The cellulase showed its maximal activity at pH 10, 70 °C. Zymography identified different xylanases produced by both molds, and some fragment sizes were highlighted: 35, 100, and 170 kDa for S. kiliense and 20, 40, 80, and 170 kDa for T. virens. In both cases, endo-1,4-β-D-xylanase activitieswere confirmed through mass spectrometry. [less ▲]

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See detailDesign of a fungal biofilm reactor for recombinant protein production from Aspergillus oryzae
Zune, Quentin ULg; Delepierre, Anissa ULg; Bauwens, Julien ULg et al

Poster (2014, September 07)

Fungi are microorganisms exhibiting high secretive power of various metabolites and have the ability to perform post-translational modifications during protein synthesis. In the field of fermentation ... [more ▼]

Fungi are microorganisms exhibiting high secretive power of various metabolites and have the ability to perform post-translational modifications during protein synthesis. In the field of fermentation industry, they are ideal hosts for secondary metabolites and recombinant protein production. At the industrial-scale, equipments usually required for solid-state or submerged fermentation of filamentous fungi have demonstrated their limitations in terms of productivity, mass transfers or products recovery (1, 2). Recently, fungal biofilm reactors were designed to combine advantages from submerged and solid-state culture and reveal their usefulness for greater secondary metabolites production relative to submerged culture conditions (3). In our work, we propose the design of a fungal biofilm reactor for a recombinant protein production from an Aspergillus oryzae strain containing a GFP reporter gene system under the control of a promoter specifically induced in solid-state conditions. The fungal biofilm reactor is composed of a metal structured packing, having the function of inert support for biofilm growth, immerged or aspersed by a liquid medium. Whereas recombinant protein production is not significantly different at the flask-scale between submerged and biofilm conditions, productivity is higher in the submerged conditions at the bioreactor-scale. Presence of recombinant proteins entrapped in the biofilm matrix highlights a diffusion constraint and a lower mass transfer in our fungal biofilm reactor. However, persistence of a free liquid biomass of low viscosity and fungal biomass retention on the support are attractive for the implementation of a continuous process in our fungal biofilm reactor. Further studies will consider a 2-D proteomic comparison of the extracellular medium from fungal biofilm reactor and submerged culture conditions in order to better understand proteins secretion and identify over-expressed proteins in biofilm conditions. [less ▲]

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See detailPropriétés physico-chimiques et biologiques des substances humiques en relation avec le développement végétal
Tahiri, Abdelghani ULg; Destain, Jacqueline ULg; Druart, Philippe et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2014), 18(3), 436-445

Les substances humiques (SH) sont des composés organiques résultant de la transformation physique, chimique et microbiologique des résidus végétaux et animaux. Elles se retrouvent sous forme de ... [more ▼]

Les substances humiques (SH) sont des composés organiques résultant de la transformation physique, chimique et microbiologique des résidus végétaux et animaux. Elles se retrouvent sous forme de macromolécules carbonées hétérogènes et complexes dans tous les écosystèmes au niveau des sols ainsi que des sédiments, des eaux de surface et des lixiviats de décharges. Elles sont issues de processus d’humification déférents, générant ainsi des molécules variables et complexes composées principalement de carbone, d’hydrogène, d’oxygène, d’azote, de soufre et de groupements fonctionnels (COOH, OH, C=O). Ces substances sont connues pour influencer les propriétés du sol et interagir avec la croissance et le développement des plantes ou avec l’activité des microorganismes. L’influence de la croissance des plantes s’exprime généralement de manière directe via la stimulation des processus biochimiques et métaboliques ou indirecte via l’amélioration de la nutrition minérale. Mais, l’intensité de réponse reste dépendante de différents paramètres tels que l’origine, la nature de la matière organique initiale, les processus de transformation et la concentration des SH ainsi que des conditions expérimentales et des plantes traitées. Les mécanismes par lesquels les SH exercent leurs effets favorables sur les végétaux sont imprécis et généralement pas bien compris. [less ▲]

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See detailMultidisciplinary approaches and fractionations to study lower termite symbiotic system and ligno-cellulose digestion
Bauwens, Julien ULg; Brasseur, Catherine ULg; Tarayre, Cédric ULg et al

Poster (2014, August)

Wood-feeding termites are a considerable source of enzymes active on ligno-cellulosic compounds. These enzymes are produced by the termite host and some representatives of its symbiotic microbial ... [more ▼]

Wood-feeding termites are a considerable source of enzymes active on ligno-cellulosic compounds. These enzymes are produced by the termite host and some representatives of its symbiotic microbial community, and are of particular interest in regard second generation biofuel. However, the complexity of microbial interactions renders micro-organisms isolation very difficult. Culture-independent methods permitted to gather a large amount of data and to understand a little more the role of each microbial population, particularly the prokaryotes. Proteomics allows working on the final product of gene expression, and corresponds more to the real operation of the digestive system. In order to investigate such a complex system, it is necessary to use multidisciplinary approaches and to fractionate this system. Zymography or affinity chromatography are used in parallel of routine proteomics techniques such as two-dimensional gel electrophoresis associated to MALDI-TOF mass spectrometry and nano-LC ESI-MS/MS. We used an artificial-diet based rearing to induce changes in microbial population balance. We performed preliminary assay to investigate the glycosylated proteome in the hindgut of a lower termite, using Multi-Lectin Affinity Chromatography (M-LAC) and enzymatic activity of harvested fractions was assessed on cellulosic substrates. [less ▲]

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See detailBiorefine: Recovery of nutrients and metallic trace elements from different wastes by chemical and biochemical processes
Tarayre, Cédric ULg; Fischer, Christophe ULg; De Clercq, Lies et al

Conference (2014, June 05)

At present, most waste processing operations are not oriented towards the valorization of valuable reusable components such as nitrogen, phosphorus, potassium and even Metallic Trace Elements (MTEs ... [more ▼]

At present, most waste processing operations are not oriented towards the valorization of valuable reusable components such as nitrogen, phosphorus, potassium and even Metallic Trace Elements (MTEs). Currently, sewage sludge, for example is usually used as a fertilizer in agriculture, in energy production or in the field of construction. Ashes originating from sludge incineration contain heavy metals and minerals in large quantities. Manure is mainly used in agriculture, although considerable amounts of nutrients are lost and cause pollution. Digestate is also used in agriculture, but other alternatives have been proposed, such as the energetic valorization. Better valorization of these wastes in agriculture (or other sectors) is however largely constrained by a multitude of legal requirements. An important problematic point is the concentration in MTEs that is found in those wastes. Consequently, recovery of nutrients and MTEsmay be a key solution for optimal valorization of wastes. Many unit operations used in the field of chemical and biochemical engineering (mechanical operations on fluids, solids, mass and heat transfers, chemical reactions, etc.) could be used in order to achieve an efficient recovery yield of nutrients and trace elements. The aim of the BioRefine Project is to make an inventory of all recovery techniques of nutrients and MTEs in five countries: Belgium, France, Germany, United Kingdom and The Netherlands. Pilot plants will also be tested to assess the efficiency of new treatment techniques after which the most efficient processes will be chosen to be applied on a larger scale. In addition, the collected data will be used to propose exploitation scenarios taking into account legal constraints and optimized logistics.This work is supported through an INTERREG IVB NWE programme(ref. 320J-BIOREFINE). [less ▲]

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See detailMALDI Mass Spectrometry Imaging: a new tool to decipher the antibiome of Bacillus amyloliquefaciens
Debois, Delphine ULg; Jourdan, Emmanuel; Cawoy, Hélène et al

Conference (2014, June 05)

Soil Bacillus isolates may devote up to 8% of their genome to nonribosomal synthesis of lipopeptide (LP)- and polyketide (PK)-type antibiotics. LPs from surfactin, iturin and fengycin families are known ... [more ▼]

Soil Bacillus isolates may devote up to 8% of their genome to nonribosomal synthesis of lipopeptide (LP)- and polyketide (PK)-type antibiotics. LPs from surfactin, iturin and fengycin families are known to exert different actions on the wellness of the producing strain such as fungitoxicity (iturin, fengycin) or motility, root colonization and immune stimulating agent (surfactin). Nevertheless, few is reported about the actual antibiome secreted in situ by Bacillus cells during confrontation with phytopathogens or plant root colonization. We developed a method mimicking the conditions prevailing in the rhizosphere and, taking advantage of the versatility of Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging, we were able to localize and identify antibiotics produced in situ by bacterial cells. First, we applied this new methodology to bioassays in which Bacillus amyloliquefaciens 98S were grown together with Fusarium oxysporum, with the aim of deciphering the role of the different LP families during the phytopathogen growth inhibition. Our results showed that the three LP families were readily produced in different proportions. Especially, images of surfactins, iturins and fengycins demonstrated that iturins are the antibiotic family actually involved in the antagonism against Fusarium oxysporum. In a second approach, we used a “in planta” model in which Bacillus amyloliquefaciens S499 was simultaneously grown with Tomato and Arabidopsis thaliana roots. Imaging results, obtained during a time course analysis, showed that surfactin is always the major lipopeptide detected. In further experiments involving a refined time-window, we observed that surfactin is actually produced as soon as 24h post inoculation. These results were the starting point of a wider study showing that the early accumulation of surfactin is a complex phenomenon involving, among other mechanisms, cell-well components recognition by bacteria, and that this interaction is a win-win association for both plant and bacterial cells. [less ▲]

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See detailResearch of phosphate accumulating microorganisms from WWTPs for the recovery of phosphorus from organic wastes (3BV.3.47)
Tarayre, Cédric ULg; Michels, Evi; Buysse, Jeroen et al

Poster (2014, June)

Many wastes containing reusable components, such as nitrogen, phosphorus and potassium, are not exploited through ideal processes. As an example, in Wallonia (Belgium), the main treatment applied to ... [more ▼]

Many wastes containing reusable components, such as nitrogen, phosphorus and potassium, are not exploited through ideal processes. As an example, in Wallonia (Belgium), the main treatment applied to sewage sludge consists in incinerating the material. Such a process is chosen when the heavy metals are too concentrated in the sludge, preventing an agricultural use. However, sewage sludge, as well as manure, slurry and digestate, contain notable amounts of nutrients (nitrogen, phosphorus and potassium). Some Waste Water Treatment Plants (WWTPs) are actually designed in order to promote Phosphate Accumulating Organisms (PAOs), able to store or release phosphorus in accordance with the environmental conditions. The aim of this work is to isolate PAOs from WWTPs and evaluate their applicability to phosphorus recovery from organic wastes. Metagenomics and metabolic properties are also considered. This work is supported by the BioRefine Project, a European project in which various member states focus on recovery of inorganics from organic wastestreams. [less ▲]

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See detailTest of humic substances on in vitro roots initiation using isolated leaves of woody species
Tahiri, Abdelghani ULg; Destain, Jacqueline ULg; Thonart, Philippe ULg et al

Poster (2014, May 15)

Arise from the chemical and biological degradation of plant and animal residues and from the synthetic activities of microorganisms in the soil, humic substances (HS) are natural heterogeneous aromatic ... [more ▼]

Arise from the chemical and biological degradation of plant and animal residues and from the synthetic activities of microorganisms in the soil, humic substances (HS) are natural heterogeneous aromatic and organic compounds. These substances are chemically complex with no clearly defined chemical structure, although generalized models have been proposed and they can be divided into fractions of humic acids, fulvic acids and humins depending on their solubility in water as a function of the pH. The stimulation of plant growth and development by HS are the activities that have attracted the attention of many scientists. They influence plant productivity directly by the stimulation of biochemical and metabolic processes or indirectly through the modification of soil characteristics and microflora activities. All together, these properties mainly affect root architecture. By inducing root hairs proliferation, differentiating root cells and enhancing lateral root emergence, an increase of the total root biomass is observed. Experiments targeting the rooting stages in absence of interferences were conducted in vitro using HS extracted from landfill leachate and a stable commercial formulation (“Humifirst” from TRADECORP company: 12% humic acid and fulvic acid 3%) issued from leonardite. Shoots and leaves explants of silver birch (Betula pendula Roth) and black alder (Alnus glutinosa L. Gaertn) vitro-plants were treated with 10 ppm leachate HS and 100 ppm Humifirst HS for 5 days during the rooting induction/initiation phase. The treated explants were then transferred into elongation medium containing only nitrate calcium for 4 weeks. The results show that application of HS during the root induction/initiation phase did not significantly influence root growth of both species in comparison with control explants. [less ▲]

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See detailEnzymatic process for the fractionation of baker’s yeast cell wall (Saccharomyces cerevisiae)
Borchani, Chema; Fonteyn, Fabienne; Jamin, Guilhem et al

in Food Chemistry (2014), 163

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See detailEffect of metal nanoparticles encapsulated in porous silica on the biphenyl biodegradation by Rhodococcus erythropolis T902.1
Wannoussa, Wissal ULg; Hiligsmann, Serge ULg; Heinrichs, Benoît ULg et al

Poster (2014, May 01)

Biphenyl is stable and highly hydrophobic, thus having a low availability for degrading microorganisms, which need an aqueous environment for their growth. As a consequence, bioremediation processes are ... [more ▼]

Biphenyl is stable and highly hydrophobic, thus having a low availability for degrading microorganisms, which need an aqueous environment for their growth. As a consequence, bioremediation processes are very limited. However, a few studies showed that using a low metal concentration accelerates the biodegradation of pollutants (Yeom and Yoo1997). Nanoparticles are considered as a new generation of compounds to improve environmental remediation and biological processes (Beckers et al. 2013; Zhang 2003). This paper investigated the enhancement effect of nanometre-sized metallic Cu, Ag, Pd or Co, on the biphenyl biodegradation by Rhodococcus erythropolis T902.1. Have been synthesized by a sol–gel process (Lambert et al. 2004). These nanoparticles (NP) of about 2–3 nm were encapsulated in porous silica (SiO2) and were added at low concentration (10-4M) in the M284 minimal medium with 500 ppm biphenyl as source of carbon and energy. The cultures containing Pd or Co produced 30% more biphenyl degraded with a higher Rhodococcus growth than those without NP (positive control) or with silica particles only. On the contrary, the presence of 10-4 M Cu or Ag nanoparticles showed an inhibitory effect on bacterial growth and biphenyl degradation compared to the positive. [less ▲]

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See detailSpatiotemporal monitoring of the antibiome secreted by Bacillus biofilms on plant roots using MALDI mass spectrometry imaging
Debois, Delphine ULg; Jourdan, Emmanuel; Smargiasso, Nicolas ULg et al

in Analytical Chemistry (2014), 86(9), 4431-4438

Some soil Bacilli living in association with plant roots can protect their host from infection by pathogenic microbes and are therefore being developed as biological agents to control plant diseases. The ... [more ▼]

Some soil Bacilli living in association with plant roots can protect their host from infection by pathogenic microbes and are therefore being developed as biological agents to control plant diseases. The plant protective activity of these bacteria has been correlated with the potential to secrete a wide array of antibiotic compounds upon growth as planktonic cells in isolated cultures under laboratory conditions. However, in situ expression of these antibiotics in the rhizosphere where bacterial cells naturally colonize root tissues is still poorly understood. In this work, we used Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI MSI) to examine spatio-temporal changes in the secreted antibiome of B. amyloliquefaciens developing as biofilms on roots. Non-ribosomal lipopeptides such as the plant immunity elicitor surfactin or the highly fungitoxic iturins and fengycins were readily produced albeit in different time-frames and quantities in the surrounding medium. Interestingly, MS/MS experiments performed directly from the gelified culture medium, also allowed to identify a new variant of surfactins released at later time points. However, no other bioactive compounds such as polyketides were detected at any time, strongly suggesting that the antibiome expressed in planta by B. amyloliquefaciens does not reflect the vast genetic arsenal devoted to the formation of such compounds. This first dynamic study reveals the power of MALDI MSI as tool to identify and map antibiotics synthesized by root-associated bacteria and more generally, to investigate plant-microbe interactions at the molecular level. [less ▲]

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See detailStochastic exposure to sub-lethal high temperature enhances exopolysaccharides (EPS) excretion and improves Bifidobacterium bifidum cell survival to freeze-drying
Nguyen, Huu Thanh ULg; Razafindralambo, Hary; Blecker, Christophe ULg et al

in Biochemical Engineering Journal (2014), 88

Exposure of microbial cells to sub-lethal stresses is known to increase cell robustness. In this work, a two-compartment bioreactor in which microbial cells are stochastically exposed to sub-lethal ... [more ▼]

Exposure of microbial cells to sub-lethal stresses is known to increase cell robustness. In this work, a two-compartment bioreactor in which microbial cells are stochastically exposed to sub-lethal temperature stresses has been used in order to investigate the response of the stress sensitive Bifidobacterium bifidum THT 0101 to downstream processing operations. A stochastic model validated by residence time distribution experiments has shown that in the heat-shock configuration, a two-compartment bioreactor (TCB) allows the exposure of microbial cells to sub-lethal temperature of 42°C for a duration comprised between 100 and 300 seconds. This exposure resulted in a significant increase of cell resistance to freeze-drying by comparison with cells cultivated in conventional bioreactors or in the TCB in the cold shock mode (CS-TCB). The mechanism behind this robustness seems to be related with the coating of microbial cells with exopolysaccharide (EPS), as assessed by the change of the zeta potential and the presence of higher EPS concentration after heat shock. Conditioning of Bifidobacteria on the basis of the heat shock technique is interesting from the practical and economical point of view since this strategy can be directly implemented in the bioreactor during stationary phase preceding cell recovery and freeze-drying [less ▲]

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See detailEnzymatic process for the fractionation of baker's yeast cell wall (saccharomyces cerevisiae)
Borchani, Chema; Fonteyn, Fabienne; Jamin, Guilhem et al

Conference (2014, April 07)

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See detailImaging MS: strategies for the identification of analytes
Debois, Delphine ULg; Smargiasso, Nicolas ULg; Jourdan, Emmanuel et al

Scientific conference (2014, April 04)

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See detailUse of on-line flow cytometry for the characterization of microbial stress dynamics during the bioprocess
Brognaux, Alison ULg; Han, Shanshan; Sorensen, Soren et al

Conference (2014, April 03)

Microbial cell population heterogeneity is now recognized as a major source of issues for the development and optimization of bioprocesses. Flow cytometry is a very powerful tool for the follow up of ... [more ▼]

Microbial cell population heterogeneity is now recognized as a major source of issues for the development and optimization of bioprocesses. Flow cytometry is a very powerful tool for the follow up of physiological properties of microbial cells in process-related conditions at the single cell level, and can be used to study the dynamics of segregation directly in bioreactors. In this context, specific interfaces have been developed in order to connect flow cytometer (FC) directly on bioreactor for automated analyses. In this work, we propose a simplified version of such interface and demonstrated its usefulness for multiplexed experiments. This automated FC system has been tested for the follow up of the dynamics of an E. coli pfis::gfpAAV fluorescent bio-reporter and its PI uptake, correlated with membrane permeability. This bioreporter is composed of a fis promoter, a growth dependent promoter-indicator of the nutrient status of cells, fused to a gene expressing an unstable variant of GFP. The results obtained showed that the dynamics of the GFP synthesis is complex and can be attributed to a complex set of biological parameters. Segregation in the membrane permeability has been noticed. This work demonstrates that a simplified version of on-line FC can be used at the process level for the investigation of the dynamics of complex physiological mechanisms. [less ▲]

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