References of "Delvigne, Frank"
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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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See detailCFD-based Compartment model for description of mixing in bioreactors
Delafosse, Angélique ULg; Calvo, Sébastien ULg; Collignon, Marie-Laure ULg et al

in Chemical Engineering Science (2014), 106

In most bioprocesses, it is fundamental to accurately predict the hydrodynamics behavior of bioreactors of different size and its interaction with the biological reaction. Computational Fluid Dynamics can ... [more ▼]

In most bioprocesses, it is fundamental to accurately predict the hydrodynamics behavior of bioreactors of different size and its interaction with the biological reaction. Computational Fluid Dynamics can provide detailed modeling about hydrodynamics and mixing. However, it is computationally intensive, especially when reactions are taken into account. Another way to predict hydrodynamics is the use of “Compartment” or “Network-of-zones” model which are much less demanding in computation time than CFD. However, compartments and fluxes between them are often defined by considering global quantities not representative of the flow complexity. To overcome the limitations of these two methods, a solution is to combine compartment modeling and CFD simulations. The aim of this study is to propose a compartment model where the flow rates between two adjacent compartments are easily computed from the velocity fields obtained by CFD. The mixing evolution predicted by the CFD-based compartment model have been then compared with mixing experiment results. Unlike a CFD mixing simulation and a classical compartment model, the CFD-based compartment model proposed in this work reproduces with a good accuracy the spatial distribution of concentrations during the mixing process and this, without any adjustable parameters. [less ▲]

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See detailScale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Brognaux, Alison ULg; Francis, Frédéric ULg; Twizere, Jean-Claude ULg et al

in Bioprocess and Biosystems Engineering (2014)

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane ... [more ▼]

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane permeability and the synthesis of several outer-membrane components allowing to cope with substrate limitation commonly found in high-cell density culture. A comparative analysis of protein leakage has thus been performed in well-mixed bioreactors and in scale-down devices. The extracellular proteome of E.coli has been investigated by 2D-gel electrophoresis and identified by subsequent MALDI-TOF analysis. On 110 picked spots, 67 proteins have been identified and the sub-localisation and the molecular function of these proteins have been determined. A majority of the extracellular proteome was composed of outer-membrane and periplasmic proteins (64%) confirming the fact that leakage is involved in high-cell density cultures. About 50% of this extracellular proteome was composed of transport and binding proteins. Furthermore, the more abundant spots on the gel corresponded to porin proteins and periplasmic transporters. In particular, the OmpC porin was found to be very abundant. Moreover, the scale-down effect on this extracellular proteome has been investigated by 2D-DIGE analysis (2-Dimensional Differential in-Gel Electrophoresis) and significant differences have been observed by comparison with culture carried out in well-mixed systems. Indeed, since substrate limitation signal is alleviated in this kind of apparatus, cell permeability was lowered as shown by flow cytometry. In scale-down conditions, protein leakage was thus less abundant. [less ▲]

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See detailScale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Brognaux, Alison ULg; Francis, Frédéric ULg; Twizere, Jean-Claude ULg et al

in Bioprocess and Biosystems Engineering (2014)

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane ... [more ▼]

Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane permeability and the synthesis of several outer-membrane components allowing to cope with substrate limitation commonly found in high-cell density culture. A comparative analysis of protein leakage has thus been performed in well-mixed bioreactors and in scale-down devices. The extracellular proteome of E.coli has been investigated by 2D-gel electrophoresis and identified by subsequent MALDI-TOF analysis. On 110 picked spots, 67 proteins have been identified and the sub-localisation and the molecular function of these proteins have been determined. A majority of the extracellular proteome was composed of outer-membrane and periplasmic proteins (64%) confirming the fact that leakage is involved in high-cell density cultures. About 50% of this extracellular proteome was composed of transport and binding proteins. Furthermore, the more abundant spots on the gel corresponded to porin proteins and periplasmic transporters. In particular, the OmpC porin was found to be very abundant. Moreover, the scale-down effect on this extracellular proteome has been investigated by 2D-DIGE analysis (2-Dimensional Differential in-Gel Electrophoresis) and significant differences have been observed by comparison with culture carried out in well-mixed systems. Indeed, since substrate limitation signal is alleviated in this kind of apparatus, cell permeability was lowered as shown by flow cytometry. In scale-down conditions, protein leakage was thus less abundant. [less ▲]

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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

Poster (2014, February 07)

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See detailIMPLEMENTATION OF A METAL STRUCTURED PACKING IN A FUNGAL BIOFILM REACTOR FOR THE PRODUCTION OF A RECOMBINANT PROTEIN BY ASPERGILLUS ORYZAE
Zune, Quentin ULg; Delepierre, Anissa; Toye, Dominique ULg et al

in Communications in Agricultural and Applied Biological Sciences (2014, February 07)

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See detailBiofilms from entomopathogenic fungi in mosquito control
Bawin, Thomas ULg; Boukraa, Slimane; Seye, Fawrou et al

Poster (2014, February 07)

Mosquitoes (Diptera: Culicidae) are zoonotic vectors of medical and veterinary importance. As part of an integrated vector control, metabolites secreted by entomopathogenic fungi could be developed as ... [more ▼]

Mosquitoes (Diptera: Culicidae) are zoonotic vectors of medical and veterinary importance. As part of an integrated vector control, metabolites secreted by entomopathogenic fungi could be developed as biopesticides. In this context, filamentous microorganisms growing on a support as biofilm in a liquid medium would offer several advantages in bioreactor regarding performances and metabolites recovery. The production of toxic metabolites by an entomopathogenic fungus Aspergillus flavus in such conditions was assessed. Three initial inoculum levels, i.e. 10^1, 10^3 and 10^6 spores/ml of PYG medium, have been tested in shake flask with or without support. Toxicity tests were performed on Culex quinquefasciatus larvae using dilutions of 1, 2, 4, 6, 8 and 10% of liquid cultures. The results indicated that A. flavus tends to form pellets in submerged culture; the size and the amount of pellets was affected by the initial inoculum level of spores. Under similar conditions, the filaments fixed on a support and didn’t appear in free form in the liquid. Toxicity tests revealed differences between both free and fixed forms. All combined conditions, LC50s ranging up to dilutions of 2.2 and 4.8% were observed within 48 hours. Secretomes could be compared between these culture conditions by proteomic and metabolomic approaches. [less ▲]

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See detailUsing micro-injection technique to assess fungal toxicity in mosquito control
Bawin, Thomas ULg; Boukraa, Slimane; Seye, Fawrou et al

in Communications in Agricultural and Applied Biological Sciences (2014, February 07), 79(1), 181-185

Topical application of insecticidal compounds allows directly exposing these substances on insect tissues and measuring their toxicity while ignoring many factors. However, this technique remains ... [more ▼]

Topical application of insecticidal compounds allows directly exposing these substances on insect tissues and measuring their toxicity while ignoring many factors. However, this technique remains difficult to apply on mosquito larvae considering their aquatic lifestyle. Micro-injection could be used for the direct deposition of toxic compounds in the larvae. Capillaries exhibiting an injection tip with an external diameter of 0.5 mm have been designed from silica tubes. For each treatment, a capillary is mounted on a pump connected to a flow rate regulator. Culex quinquefasciatus larvae were injected with 10^7 spores/ml of entomopathogenic fungi (Aspergillus clavatus, Metarhizium anisopliae, Metarhizium sp.). Mortalities were recorded daily during 72h. The distribution of spores stained with methylene blue and injected into the body of larvae was also observed according to the system described. Results showed that spores were distributed over the whole body. The injection of Aspergillus clavatus, Metarhizium anisopliae and Metarhizium sp spores induced corrected mortalities of 62%, 53% and 57% after 72h, and differed statistically from control groups. Finally, post-mortem emergences of filaments from dead larvae were observed in the case of the three fungal strains confirming spore viability. Injection of inactivated spores (or inert bodies of similar size) could help to reject the hypothesis of a response due to the presence of foreign bodies. [less ▲]

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See detailThe role of protein modifications in senescence of freeze-dried Acetobacter senegalensis during storage
Shafiei, Rasoul ULg; Zarmehrkhorshid, Raziyeh ULg; Bentaib, Azeddine ULg et al

in Microbial Cell Factories (2014)

Background Loss of viability is one of the most important problems during starter culture production. Previous research has mostly focused on the production process of bacterial starters, but there are ... [more ▼]

Background Loss of viability is one of the most important problems during starter culture production. Previous research has mostly focused on the production process of bacterial starters, but there are few studies about cellular protein deterioration causing cell defectiveness during storage. In the present study, we investigated the influence of storage temperature (−21, 4, 35°C) on the cellular protein modifications which may contribute to the senescence of freeze-dried Acetobacter senegalensis. Results Heterogeneous populations composed of culturable cells, viable but non-culturable cells (VBNC) and dead cells were generated when freeze-dried cells were kept at −21 and 4°C for 12 months whereas higher storage temperature (35°C) mainly caused death of the cells. The analysis of stored cell proteome by 2D-DiGE demonstrated a modified pattern of protein profile for cell kept at 4 and 35°C due to the formation of protein spot trains and shift of Isoelectric point (pI). Quantification of carbonylated protein by ELISA showed that the cells stored at 4 and 35°C had higher carbonylated protein contents than fresh cells. 2D-DiGE followed by Western blotting also confirmed the carbonylation of cellular proteins involved in translation process and energy generation. The auto-fluorescent feature of cells kept at 35°C increased significantly which may be an indication of protein glycation during storage. In addition, the percentage of cellular unsaturated fatty acid and the solubility of cellular proteins decreased upon storage of cells at higher temperature suggesting that peroxidation of fatty acids and possibly protein lipidation and oxidation occurred. Conclusions High storage temperature induces some deteriorative reactions such as protein oxidation, lipidation and glycation which may cause further protein modifications like pI-shift, and protein insolubility. These modifications can partly account for the changes in cell viability. It can also be deduced that even moderate carbonylation of some critical cellular proteins (like ribosomal proteins) may lead to VBNC formation or death of freeze-dried bacteria. Moreover, it seems that other mechanisms of biomolecule deterioration preceding protein carbonylation lead to VBNC formation under very low storage temperature. [less ▲]

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See detailMicrobial heterogeneity affects bioprocess robustness: Dynamic single cell analysis contribute to understanding microbial populations
Delvigne, Frank ULg; Goffin, Philippe

in Biotechnology Journal (2014), 9(1), 61-72

Heterogeneity or segregation of microbial populations has been the subject of much research, but the real impact of this phenomenon on bioprocesses remains not well understood. The main reason behind this ... [more ▼]

Heterogeneity or segregation of microbial populations has been the subject of much research, but the real impact of this phenomenon on bioprocesses remains not well understood. The main reason behind this lack of knowledge is the difficulty for monitoring microbial population heterogeneity in dynamic process conditions. The main concepts leading to microbial population heterogeneity in the context of bioprocesses have been summarized by two distinct hypotheses. The first one involves the individual history of microbial cells or “path” followed during their residence time inside process equipment. The second one involves a coordinated response of the microbial population as a bet-hedging strategy in order to cope with process-related stresses. The respective contribution of each hypothesis to microbial heterogeneity in bioprocesses is still unclear. This statement illustrates the fact that, although microbial phenotypic heterogeneity has been thoroughly investigated at the fundamental level, the implications of this phenomenon in the context of microbial bioprocesses are still subjected to debate. At this time, automated flow cytometry is the best technique for the investigation of microbial heterogeneity in process conditions. However, dedicated software and relevant biomarkers are needed for its proper integration as a bioprocess control tool. [less ▲]

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See detailBioRefine: Recovery of Nutrients from Sewage Sludge, Manure and Digestate by a Combination of Chemical and Biochemical Unit Operations
Tarayre, Cédric ULg; Delvigne, Frank ULg; Michels, Evi et al

Poster (2013, December)

At this time, many wastes are exploited through processes that do not really consider applications potentially more profitable. Such wastes contain reusable components, such as nitrogen, phosphorus and ... [more ▼]

At this time, many wastes are exploited through processes that do not really consider applications potentially more profitable. Such wastes contain reusable components, such as nitrogen, phosphorus and potassium, whereas heavy metals may also be considered. Their composition depends on input materials, and considerable heterogeneities must be highlighted. Sewage sludge is usually exploited as a fertilizer in agriculture, in energy production or in the field of construction. The main application of manure is 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 combustion. The use of waste in agriculture must respect many legal constraints. Another problematic point is the concentration of heavy metals that is found in those wastes. Consequently, recovery of nutrients and trace elements may be a key solution. Chemical and biochemical engineering propose many unit operations (mechanical operations on fluids, solids, mass and heat transfers, chemical reactions, etc.) that may be used to reach an efficient recovery yield of capital nutrients and trace elements. Here, we propose a methodology which consists in considering the unit operations separately with their own input and output flows, energy and heat consumption, investment, etc. and combining them to simulate industrial processes. The element concentrations and their forms will also be considered using a classification matrix. Combinations of unit operations will lead to reliable processes that should be applied on an industrial scale. This work is supported by the BioRefine Project, a European project in which various member states focus on recovery of inorganics from organic waste streams. We gratefully acknowledge the INTERREG IVB NWE programme, which financed the BioRefine Project (ref. 320J-BIOREFINE). [less ▲]

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

Poster (2013, November 15)

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See detailPrésentation des projets Interreg IVB BIOREFINE et RENEW
Tarayre, Cédric ULg; Delvigne, Frank ULg; Destain, Jacqueline ULg et al

Conference (2013, October 22)

Les projets BioRefine et Renew sont financés par le programme de coopération européen Interreg IVB. Leur but est de promouvoir les transferts de compétences, d'informations et de matières entre les ... [more ▼]

Les projets BioRefine et Renew sont financés par le programme de coopération européen Interreg IVB. Leur but est de promouvoir les transferts de compétences, d'informations et de matières entre les différents pays de la zone européenne du Nord-Ouest. Le projet BioRefine a pour but de récupérer les nutriments (N, P et K) ainsi que les éléments traces métalliques à partir de déchets bien spécifiques: le fumier, le lisier, les digestats de méthanisation et les boues de stations d'épuration. Cette récupération requiert l'élaboration de procédés industriels mis en place après une étude en laboratoire. Le projet Renew, lui, est beaucoup plus général. En ce qui concerne le rôle de Gembloux Agro-Bio Tech dans le projet, le travail sera essentiellement focalisé sur la production d'acide succinique à partir de déchets. Cette production sera assurée par la souche bactérienne Actinobacillus succinogenes. [less ▲]

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