References of "Thonart, Philippe"
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See detailPrise en compte l'état de la membrane cellulaire sur la réponse de biocapteurs fluorescents pour la détection de défaut d'écoulement au niveau des bioréacteurs : vers une intégration du "sécrétome" dans la problématique de l'extrapolation
Delvigne, Frank ULg; Brognaux, Alison ULg; Gorret, Nathalie et al

Conference (2011, November 29)

Malgré les nombreuses approches envisagées à ce jour, la dynamique du stress microbien en conditions de culture intensive (procédé fed-batch) est encore un aspect mal maîtrisé. Au cours de ce travail ... [more ▼]

Malgré les nombreuses approches envisagées à ce jour, la dynamique du stress microbien en conditions de culture intensive (procédé fed-batch) est encore un aspect mal maîtrisé. Au cours de ce travail, deux biocapteurs microbiens basés sur le principe de la protéine fluorescente verte (Green Fluorescent Protein ou GFP) ont été mis en œuvre dans des réacteurs scale-down permettant de reproduire les défauts d'écoulement rencontrés dans les bioréacteurs industriels. Les promoteurs de stress associés à la synthèse de la GFP ont été choisis suivant leur sensibilité à la limitation en source de carbone, condition standard qui est rencontrée dans les processus fed-batch ou l'apport en substrat carboné est limité afin d'éviter des déviations du métabolisme microbien (promoteurs rpoS et csiE). Les résultats obtenus montrent clairement que l'exposition des biocapteurs à des hétérogénéités locales de substrat entraîne une diminution de l'expression de la GFP. L'intensité de fluorescence a été mesurée à l'échelle cellulaire par cytométrie en flux. Durant les cultures, une chute significative du niveau de GFP intracellulaire a été observée pour les deux conditions scale-down considérées et pour les deux types de promoteur. Cette chute de fluorescence peut être attribuée à des phénomènes de répression des promoteurs suite à la levée locale de la limitation en carbone, mais également au relargage de la GFP dans le milieu extracellulaire. Ce relargage a été observé dans toutes les conditions opératoires considérées, comme le montrent les analyses des surnageants de culture par SDS-PAGE. L'intensité du relargage est néanmoins plus forte dans les conditions standard de culture (c'est-à-dire dans des réacteurs classiques sans approche scale-down). En effet, la coloration par l'iodure de propidium des biocapteurs cultivés en conditions standards est plus élevée que dans les conditions scale-down, suggérant une perméabilité membranaire plus élevée. Ces résultats offrent des potentialités intéressantes pour l'analyse simultanée de la viabilité cellulaire et de la détection des défauts d'écoulement par l'emploi de biocapteurs microbiens. De plus, les analyses par SDS-PAGE ont montre une grande diversité de protéines relargée en cours de procédé. Cette observation est étonnante du fait de l'emploi de milieux minéraux définis dans toutes les conditions étudiées. L'analyse de ce sécrétome offre des potentialités intéressantes pour la caractérisation des conditions de stress encourues par les micro-organismes au cours des procédés. [less ▲]

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See detailEvolution de la viabilité cellulaire dans les procédés de production de ferments lactiques : effet du type de bactérie sur l'évaluation par cytométrie en flux
Delvigne, Frank ULg; Thiry, Christophe ULg; Pierart, Céline ULg et al

Conference (2011, November 29)

La cytométrie en flux est une technique permettant d'effectuer des analyses au niveau des cellules individuelles. Cette technique a été développée à l'origine pour l'analyse de cellules animales dans des ... [more ▼]

La cytométrie en flux est une technique permettant d'effectuer des analyses au niveau des cellules individuelles. Cette technique a été développée à l'origine pour l'analyse de cellules animales dans des applications à finalité médicale. Ce n'est que dans les années 90 que cette technique a été considérée dans le cadre des cellules microbiennes procaryotes et eucaryotes. Grâce à l'évolution des technologies (laser à l'état solide, progrès dans la maîtrise des micro-écoulements,…), plusieurs développeurs proposent actuellement des cytomètres en flux pour des budgets abordables (environ 30.000 euros). Cette évolution entraîne actuellement un engouement des industriels pour la mise en oeuvre de cette technique pour le suivi de l'évolution des populations microbiennes dans divers procédés : brasserie, vinaigrerie, production de starters pour la boulangerie, production de starters lactiques,… L'intérêt manifesté pour cette technique est simple à comprendre si on considère les techniques actuelles pour l'estimation de la viabilité cellulaire. En effet, celles-ci reposent souvent sur la revification des cellules sur milieu gélosé et comptage des colonies après un temps d'incubation. Le problème fondamental de cette technique repose sur l'utilisation de conditions de culture qui ne sont pas forcément identiques à celle rencontrée au niveau du processus de production. Ce phénomène entraîne une sous-estimation des cellules microbiennes actives rassemblées sous le terme de "viables mais non cultivables". A cela s'ajoutent des problèmes techniques imposés par le temps de remise en culture et l'obtention des résultats bien après que le processus de production soit terminé, ainsi que par des procédures de laboratoire coûteuses en personnel et en consommables. La cytométrie s'impose donc comme une technique de choix qui permet d'analyser les cellules microbiennes individuelles sans étape de remise en culture et avec un débit expérimental élevé. En effet, 30.000 cellules peuvent être analysées en 30 secondes immédiatement après la prise d'échantillon, ce qui permet éventuellement de corriger les conditions de procédés en fonction de l'état des micro-organismes. L'utilisation de fluorochromes spécifiques permet l'analyse de caractéristiques cellulaires. Dans le cas des bactéries lactiques, l'utilisation du couple de colorant "carboxyfluorescéine diacétate" / "iodure de propidium" (cFDA/PI) est mis en oeuvre en routine pour la détermination de la viabilité cellulaire. L'iodure de propidium pénètre dans les cellules ayant une membrane endommagée et les colore en rouge, tandis que le cFDA est un composé non fluorescent qui diffuse au travers de toutes les membranes cellulaires et est hydrolysées par les activités estérases intracellulaires pour donner un composé fluorescent vert. Ces deux composantes de fluorescence peuvent être facilement caractérisées par analyse multi paramétrique au niveau d'un cytomètre en flux. La difficulté majeure que rencontre l'application de la cytométrie en flux au niveau industriel réside dans l'interprétation des résultats. Dans ce travail, trois souches de bactéries lactiques ayant des sensibilités différentes au stress de procédé ont été mise en oeuvre dans des schémas de production industriels faisant intervenir les étapes suivantes : production en bioréacteur agité de 2m³, récolte par centrifugation continue, congélation et lyophilisation. L'analyse par cytométrie en flux montre des tendances fondamentalement différentes pour les trois types de micro-organismes. Les deux souches microbiennes plus sensibles au stress de procédé du fait de leur caractère anaérobie strict ou microaérophile montrent des sous-populations bien distinctes au niveau des cytogrammes, tandis que la souche plus résistante ne montre qu'une seule population évoluant suivant les étapes du procédé. Ces observations sont utilisées pour une meilleure interprétation des cytogrammes pour l'estimation de la viabilité cellulaire en conditions de procédé. Une meilleure interprétation peut également être obtenue en considérant la possibilité de relargage des produits de dégradation fluorescents provenant de l'hydrolyse du cFDA en fonction de l'état de la membrane cellulaire. [less ▲]

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See detailDetermination of optimum conditions for preservation and reactivation of freeze-dried Acetobacter senegalensis used as vinegar starter
Shafiei, Rasoul ULg; Thonart, Philippe ULg

Poster (2011, November)

Availability of efficient starters is one of the most important elements during fermentation technology. Acetic acid bacteria (AAB) face extreme conditions (low pH, high acetic acid concentration (>7 ... [more ▼]

Availability of efficient starters is one of the most important elements during fermentation technology. Acetic acid bacteria (AAB) face extreme conditions (low pH, high acetic acid concentration (>7%), ethanol and high temperature) during vinegar fermentation. In spite of introduction of broth media contained viable AAB as starter by some companies; traditional methods for initiation of vinegar fermentation are still used even in modern factories, therefore there is a long Lag phase for initiation of fermentation. In recent studies in CWBI, a kind of lyophilized starter has been produced by using an isolated thermotolerant bacterium: Acetobacter senegalensis. The aim of this study is to determine the best media for revitalization and the optimum temperature for preservation of the freeze-dried cells. To produce biomass, glucose was used as carbon source in batch culture under regulated conditions. The cells harvested when they reached to stationary phase. Harvested cells suspended in spent growth medium (final supernatant of fermentation). Maltodextrin (10%), manitol (20%) and spent growth medium were used as protestants. After freeze-drying they were subjected to storage test at -20° C, 4° C and 35° C for six months. The viability of cells determined by using spread plate technique using three different media: YG (yeast extract, glucose) and YGE (yeast extract, glucose, ethanol) and YGEA (yeast extract, glucose, ethanol, acetic acid). According to residual viable cells, manitol and maltodextrin showed higher protective functions rather than spent growth medium (92.3%, 88.2% and 82.1% survival, respectively) during freeze-drying process. Viability of cells during rehydration is completely dependent to the composition of the broth media used for this purpose. Adding ethanol (>0.5%) or acetic acid (>1%) to rehydration medium caused 40-45% reduction in viable cell numbers in comparison to YG broth. The composition of culture media can also influence the growth of bacteria after rehydration. In YG, much higher cell growth (about 1 log unit) was observed in comparison to YGE and YGEA. Addition of glucose (20 g/l) to YGE can neutralize the adverse effect of ethanol considerably but it cannot improve the growth in YGEA. This can be partially explained according to the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase. It was revealed that freeze-dried cells had very low activities for these two enzymes. Analysis of total soluble protein contents of lyophilized cells during storage at different conditions revealed that the soluble protein content of cells reduced by increasing the storage temperature. At 35° C after 15 days, there were 4.87 log units reduction in the viability of cells, and also about 14% less soluble protein was detectable under this condition whereas keeping of cells at -20 or + 4° C had no influences on protein and viability of cells. Storage of cells at +4° C in darkness for six month showed that about 79.2% and 68.3% viability of cells remained for cells protected by manitol and maltodextrin, respectively. To assess the real activity of lyophilized cells as starters, they were introduced to broth media (YGE contained 20g/l of glucose). It was seen that 6 months old lyophilized cells (with maltodextrin or manitol as protectants) were able to grow and consume 5 v/v of ethanol in 48 hr in the presence of initial acetic acid concentration (0.5%) at 30° C. In conclusion, the mentioned methods for preservation and reactivation of freeze-dried Acetobacter senegalensis can provide a promising tool for decreasing the lag phase of vinegar fermentation. [less ▲]

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See detailUse of Bacillus subtilis S499 to control mould growth during malting of red sorghum from the D.R. of Congo
Bwanganga Tawaba, Jean-Claude ULg; Destain, Jacqueline ULg; Bera, François ULg et al

Poster (2011, November)

In this work, moulds growth was monitored during malting of a red sorghum from D.R. of Congo. Results obtained with chemical treatments [diluted alkalines: NaOH 0.2% (T1) and Ca(OH)2 0.1% (T2)] were ... [more ▼]

In this work, moulds growth was monitored during malting of a red sorghum from D.R. of Congo. Results obtained with chemical treatments [diluted alkalines: NaOH 0.2% (T1) and Ca(OH)2 0.1% (T2)] were compared with those of biocontrols [use of Bacillus subtilis S499 108 cells / mL: supernatant + cells (T3), cells (T4) and cells-free supernatant (T5)]. Chemical treatments have shown the highest fungal contamination levels, the highest malting loss, but better enzyme activities (α and β-amylases, limit-dextrinase, α-glucosidase and β-glucanase). The inhibition tests showed that the concentration of lipopeptides is correlated with the reduction of moulds growth. Malting trials with starters of Bacillus subtilis S499 showed that culture dilution affects the growth of moulds during red sorghum malting. The combination of a chemical treatment such as diluted NaOH with a biocontrol could be used to improve sorghum malts quality in terms of different enzymes activities and by reducing the level of fungal contamination. Key words: Bacillus subtilis S499, mould, sorghum malting [less ▲]

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See detailFermentative biohydrogen production in a novel biodisc bioreactor: Principle and Improvement
Beckers, Laurent ULg; Hiligsmann, Serge ULg; Masset, Julien ULg et al

in Bozhou, Li (Ed.) Low Carbon Earth Summit 2011 Proceeding (2011, October 23)

In order to produce hydrogen at high yields and production rates, the biotechnological process needs to be further optimized and efficient bioreactors must be designed. A biodisc bioreactor has been ... [more ▼]

In order to produce hydrogen at high yields and production rates, the biotechnological process needs to be further optimized and efficient bioreactors must be designed. A biodisc bioreactor has been design and investigated to produce biohydrogen from glucose by the Clostridium butyricum CWBI1009 strain at a high yield and production rate. This reactor, working continuously, has an internal volume of 2.3l but a working volume (liquid phase) of 300ml. Firstly, it enhances the hydrogen production rate (by about 3 times more than a completely stirred bioreactor) by partially fixing the bacteria on the porous support and thus increasing the cell concentration in the bioreactor (decoupling of HRT and SRT). Secondly, the rotating biodisc design enables efficient gas transfer (hydrogen and carbon dioxyde) from the liquid phase where it is produced by the bacteria to the headspace. Indeed, this is an important way to increase hydrogen production yields (by about 25% compared to a completely stirred bioreactor) by allowing the bacteria to focus on the metabolites pathways that produce more hydrogen. Other reactors designs have shown such good results by increasing the interfacial surface. [less ▲]

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See detailChanges in termites feeding diets for gut micro-organisms selection and further cultivation
Bauwens, Julien ULg; Brasseur, Catherine ULg; Matteotti, Christel ULg et al

Poster (2011, October 02)

Termites gut may overcome important dietary perturbations, initial diversity acting as key point buffering effects on host, although termites possess their own enzymatic system. Some artificial diets ... [more ▼]

Termites gut may overcome important dietary perturbations, initial diversity acting as key point buffering effects on host, although termites possess their own enzymatic system. Some artificial diets permitted a simplification of the lower termites gut symbiotic system, which could be used as first step in symbionts isolation and cultivation. Preliminary assay of cultivation actually gave encouraging results. Proteomic proved to be suitable tool to investigate such a complex system. Nevertheless, for some symbionts very few genes are sequenced, which should lead to more targeted proteomic studies. Protein chromatography will allow to split up the proteome and more accurate analysis. [less ▲]

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See detailPotential use of GFP microbial biosensors for the detection of mixing imperfections and cell viability in bioreactors
Delvigne, Frank ULg; Delafosse, Angélique ULg; Collignon, Marie-Laure ULg et al

Conference (2011, September 25)

The dynamics of microbial stress response in intensive cultivation conditions remains misunderstood. In this work, two green fluorescent protein (GFP) transcriptional reporters have been used as ... [more ▼]

The dynamics of microbial stress response in intensive cultivation conditions remains misunderstood. In this work, two green fluorescent protein (GFP) transcriptional reporters have been used as biosensors of the heterogeneities generated in a two-compartment scale-down reactor. The stress promoters have been chosen for their responsiveness to carbon limitation corresponding to the global substrate profiles encountered in intensive fed-batch cultures. From our results, it can be concluded that the exposure of microbial cells to substrates heterogeneities tends to decrease the GFP expression level in fed-batch mode. Fluorescence intensities have been monitored at the single cell level by using flow cytometry. During the course of the fed-batch culture, a drop at the level of the intracellular GFP content has been observed for the two scale-down operating conditions and for the two promoters sensitive to substrate limitation (rpoS and csiE). The fluorescence drop can be attributed to the repression of these promoters but also to the release of GFP to the extracellular medium according to the increase of the fluorescence level of the supernatant. This leakage has been observed for all the operating conditions, i.e. the scale-down reactors and the culture operating in the normal mode. Interestingly, GFP leakage is more pronounced in the case of the cultures operated in the normal mode. Indeed, staining by propidium iodide (PI) tends to be more elevated for the microbial cells cultured under the normal mode by comparison with those cultured in scale-down conditions, indicating a higher permeability of the membrane. These results are in accordance with previously published ones (Hewitt and co-workers) suggesting that microbial cells cultivated in heterogeneous bioreactors (scale-down and large-scale bioreactors) exhibits a higher viability level. These results suggest that GFP microbial biosensors could be used to detect simultaneously mixing imperfections and their impact on the viability of microorganisms. [less ▲]

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See detailMICRO-H2 – Microbiological production of hydrogen: study of microalgal and bacterial processes
Calusinska, Magdalena ULg; Joris, Bernard ULg; Wilmotte, Annick ULg et al

Poster (2011, September 07)

The project MICRO-H2 aims to study and exploit the microbial (bacterial and algal) production of hydrogen (H2). In addition to building a competence centre around the H2 production by microorganisms and ... [more ▼]

The project MICRO-H2 aims to study and exploit the microbial (bacterial and algal) production of hydrogen (H2). In addition to building a competence centre around the H2 production by microorganisms and the molecular monitoring of the processes, this project tries to address two main socio-economic issues. First, transport and many economic activities will be based on hydrogen energy in the near future. Secondly, many researches and technology developments deal with renewable resources. Therefore, a new integrated technology for a sustainable development should be promoted. Photofermentation and dark-fermentation are the most promising ways to produce biohydrogen. The main advantage of the first process is the complete conversion of substrate, if any, to hydrogen. However, present H2-production rates by microalgae remain low. Therefore, a better understanding of the microalgal hydrogen metabolism and rate improvements by genetic engineering are needed. On the other hand, dark-fermentation achieves at present far higher H2-production rates, but improvements are expected through monitoring and optimisation of bacterial diversity and activity. The objectives about bacterial H2 production were to increase knowledge, stability potentialities and investigation skills about the consortia of bacteria involved in bioreactors treating wastewater rich in carbohydrates to produce biohydrogen. The project focused mainly on the study of the potentialities of different consortia, with a focus on Clostridium strains. Concerning the microalgal production of H2, the objectives were to increase knowledge on the metabolic interactions that determine H2 evolution at the cellular level and to produce new strains with increased ability for H2 production in the two-stage process. [less ▲]

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See detailMicroorganisms from aphid honeydew attract natural enemies and tending ants
Verheggen, François ULg; Leroy, Pascal; Fischer, Christophe ULg et al

Conference (2011, August)

Aphids are some of the most serious pests of cultivated crops worldwide, causing major yield and economic losses. Previous works have demonstrated ants and natural enemies (including ladybeetles and ... [more ▼]

Aphids are some of the most serious pests of cultivated crops worldwide, causing major yield and economic losses. Previous works have demonstrated ants and natural enemies (including ladybeetles and hoverflies) to be able to use aphid volatile chemicals to locate aphid colonies. Here, we report the first isolation of a bacterium from the pea aphid Acyrthosiphon pisum honeydew, Staphylococcus sciuri, which produces kairomones used by the aphidophagous hoverfly Episyrphus balteatus and the Asian Ladybeetle Harmonia axyridis during their search for prey colonies. Some specific semiochemicals produced by S. sciuri were identified as attractants and ovipositional stimulants. Similarly, we have shown scouts of the aphid tending ant species, Lasius niger, to orientate their foraging behaviour toward an Aphis fabae infested plant and we have demonstrated that the odours released by this aphid honeydew were attractive for ant scouts. Again, bacteria were involved in the production of these honeydew semiochemicals. Interestingly, ant scouts were also able to discriminate honeydew odour from A. fabae (usually attended by L. niger) and A. pisum (unattendedby L. niger). Comparison of the volatile and bacteria composition of both aphid species honeydew were attended. [less ▲]

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See detailEarthworms use odor cues to locate and feed on microorganisms in soil
Zirbes, Lara ULg; Mark, Mescher; Vrancken, Véronique ULg et al

in PLoS ONE (2011), 6(7), 21927

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See detailMicroorganisms from Aphid Honeydew Attract and Enhance the Efficacy of Natural Enemies
Leroy, Pascal ULg; Sabri, Ahmed ULg; Heuskin, Stéphanie ULg et al

in Nature Communications (2011), 2

Aphids are one of the most serious pests of crops worldwide, causing major yield and economic losses. To control aphids, natural enemies could be an option but their efficacy is sometimes limited by their ... [more ▼]

Aphids are one of the most serious pests of crops worldwide, causing major yield and economic losses. To control aphids, natural enemies could be an option but their efficacy is sometimes limited by their dispersal in natural environment. Here we report the first isolation of a bacterium from the pea aphid Acyrthosiphon pisum honeydew, Staphylococcus sciuri, which acts as a kairomone enhancing the efficiency of aphid natural enemies. Our findings represent the first case of a host-associated bacterium driving prey location and ovipositional preference for the natural enemy. We show that this bacterium has a key role in tritrophic interactions because it is the direct source of volatiles used to locate prey. Some specific semiochemicals produced by S. sciuri were also identified as significant attractants and ovipositional stimulants. The use of this host-associated bacterium could certainly provide a novel approach to control aphids in field and greenhouse systems. [less ▲]

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See detailStress condition during acetic acid fermentation
Shafiei, Rasoul ULg; Thonart, Philippe ULg

Speech/Talk (2011)

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See detailA semiochemical enhancing the attractiveness of aphidophagous predators in potato crops
Leroy, Pascal ULg; Vandereycken, Axel ULg; Sabri, Ahmed ULg et al

Poster (2011, May 24)

Aphids are major pests of crops worldwide and the use of pesticides has led to resistant populations. The integration of aphid natural enemies in integrated management programs could be an option, but ... [more ▼]

Aphids are major pests of crops worldwide and the use of pesticides has led to resistant populations. The integration of aphid natural enemies in integrated management programs could be an option, but their efficacy is often limited by their quick dispersal from the ecosystem where they are released. Here, using wind-tunnel and field experiments, we have demonstrated that 3-Methyl-2-butenal acts as an efficient attractant and ovipositional stimulant for the hoverfly Episyrphus balteatus (De Geer) (Diptera: Syrphidae), enhancing its efficiency as biological control agent in crop fields. Wind-tunnel assays allowed determining that a minimal dose of 250 µg of 3-Methyl-2-butenal is required to attract the hoverflies over a distance of 2.5 m while a minimal dose of 500 µg is needed to induce the oviposition. Both attraction and oviposition increased proportionally to the tested doses showing that hoverflies are able to regulate their oviposition according to the chemical stimuli from their environment. In field experiments, both Syrphidae and Chrysopidae were strongly attracted by the 3-Methyl-2-butenal at a dose of 10 mg which also induces the oviposition of these two aphid predators in potato crops. This study also highlighted that hoverflies are the most abundant aphid predators found in natural environment. Against aphids, the use of this semiochemical could certainly enhance the efficiency of natural enemies. Furthermore, the use of the chemical cue 3-Methyl-2-butenal could provide a novel approach to control aphids in field and greenhouse systems. [less ▲]

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See detailPotentialités d’application des technologies biologiques pour la depollution des sols en Wallonie
Aldric, Jean-Marc ULg; Druart, P.; Maesen, Philippe ULg et al

in Journal des Ingénieurs (Le) (2011), 132

Detailed reference viewed: 79 (28 ULg)