Publications of Daniel Portetelle
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See detailLa souris, le patient, et le faux expert. Décryptage d'une mystification.
Bakker, Julie ULiege; Balthazart, Jacques ULiege; Baron, Frédéric ULiege et al

Article for general public (2018)

La recherche sur animaux est actuellement encadrée de façon stricte en Wallonie comme dans toute l'Union Européenne (voir l'article de Marc Vandenheede publié dans le Vif). Cette législation et les ... [more ▼]

La recherche sur animaux est actuellement encadrée de façon stricte en Wallonie comme dans toute l'Union Européenne (voir l'article de Marc Vandenheede publié dans le Vif). Cette législation et les contrôles qui y sont associés induisent de nombreuses contraintes pratiques, des charges administratives et des coûts financiers importants que les chercheurs seraient certainement heureux d'éviter s'il existait une alternative à l'expérimentation animale. [less ▲]

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See detailReducing agent can be omitted in the incubation medium of the batch in vitro fermentation model of the pig intestines
Poelaert, Christine; Nollevaux, Geraldine; Boudry, Christelle ULiege et al

in Animal (2018), 12

Over the past decade, in vitro methods have been developed to study intestinal fermentation in pigs and its influence on the digestive physiology and health. In these methods, ingredients are fermented by ... [more ▼]

Over the past decade, in vitro methods have been developed to study intestinal fermentation in pigs and its influence on the digestive physiology and health. In these methods, ingredients are fermented by a bacterial inoculum diluted in a mineral buffer solution. Generally, a reducing agent such as Na2S or cysteine-HCl generates the required anaerobic environment by releasing metabolites similar to those produced when protein is fermented, possibly inducing a dysbiosis. An experiment was conducted to study the impact of two reducing agents on results yielded by such in vitro fermentation models. Protein (soybean proteins, casein) and carbohydrate (potato starch, cellulose) ingredients were fermented in vitro by bacteria isolated from fresh feces obtained from three sows in three carbonate-based incubation media differing in reducing agent: (i) Na2S, (ii) cysteine-HCl and (iii) control with a mere saturation with CO2 and devoid of reducing agent. The gas production during fermentation was recorded over 72 h. Short chain fatty acids (SCFA) production after 24 and 72 h and microbial composition of the fermentation broth after 24 h were compared between ingredients and between reducing agents. The fermentation residues after 24 h were also evaluated in terms of cytotoxicity using Caco-2 cell monolayers. Results showed that the effect of the ingredient induced higher differences than the reducing agent. Among the latter, cysteine-HCl induced the strongest differences compared with the control, whereas Na2S was similar to the control for most parameters. For all ingredients, final gas produced per g of substrate was similar ( P>0.10) for the three reducing agents whereas the maximum rate of gas production ( Rmax) was reduced ( P<0.05) when carbohydrate ingredients were fermented with cysteine-HCl in comparison to Na2S and the control. For all ingredients, total SCFA production was similar ( P>0.10) after 24 h of fermentation with Na2S and in the control without reducing agent. Molar ratios of branched chain-fatty acids were higher ( P<0.05) for protein (36.5% and 9.7% for casein and soybean proteins, respectively) than for carbohydrate (<4%) ingredients. Only fermentation residues of casein showed a possible cytotoxic effect regardless of the reducing agent ( P<0.05). Concerning the microbial composition of the fermentation broth, most significant differences in phyla and in genera ascribable to the reducing agent were found with potato starch and casein. In conclusion, saturating the incubation media with CO2 seems sufficient to generate a suitable anaerobic environment for intestinal microbes and the use of a reducing agent can be omitted. [less ▲]

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See detailEffects of protein source and cooking procedure on intestinal microbiota and on fermentation end-products in rats
POELAERT, Christine ULiege; Despret, Xavier; Portetelle, Daniel ULiege et al

Poster (2016, June)

Animal and plant proteins are major proteins sources in the human diet. After their enzymatic degradation in the upper gastro-intestinal tract, the undigested fraction of these proteins is available for ... [more ▼]

Animal and plant proteins are major proteins sources in the human diet. After their enzymatic degradation in the upper gastro-intestinal tract, the undigested fraction of these proteins is available for fermentation by the microbiota of the large intestine leading to the production of short-chain fatty acids (SCFA), branched-chain fatty acids (BCFA), ammonia, biogenic amines, sulphur metabolites, phenols and indoles. As some of these compounds have genotoxic and cytotoxic effects, protein fermentation is considered as detrimental to the host’s epithelial health. BCFA are usually used as a marker of intestinal protein fermentation. We studied in vivo the impact of proteins from animal and plant origin, raw or after a cooking procedure, on the composition of gut microbiota and on fermentation end-products. Weanling rats were used as models of the human gut microbiota. Eight experimental diets were formulated with beef meat (Longissimus dorsi), chicken meat (Pectoralis major), white pea beans (Phaseolus vulgaris), soybeans (Glycine max), used raw and cooked, as sole source of protein in the diet. One casein diet was used as control. All diets, formulated to contain 15% of raw protein, were given to seven rats for four weeks. After euthanasia, caecal contents were collected. Pyrosequencing analyses (Roche 454 GS Junior Genome Sequencer) were performed to study the microbial composition. SCFA and BCFA were measured using HPLC (Waters 2690). Microbial composition in the caecum is associated to the type of dietary protein and to the cooking procedure applied. The proportion of BCFA in the caecal content is mainly affected by the type of protein. So BCFA represent respectively 04-06% and 35-44% of total SCFA with diets based on plant and on animal proteins. In conclusion, both the type of protein and the cooking procedure could impact the gut microbiota in terms of composition and of fermentative capacity. [less ▲]

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See detailIn vitro evaluation of fermentation characteristics of two types of insects as potential novel protein feeds for pigs
POELAERT, Christine ULiege; Beckers, Yves ULiege; Despret, Xavier et al

in Journal of Animal Science (2016), 94(7S3), 198-201

Novel protein sources such as insects are suggested for pig nutrition. Protein availability might be impacted by the nature of the insect and by the thermal treatment applied to sanitize this ingredient ... [more ▼]

Novel protein sources such as insects are suggested for pig nutrition. Protein availability might be impacted by the nature of the insect and by the thermal treatment applied to sanitize this ingredient. Their influence on protein availability and colonic fermentation is unknown. Plant proteins (beans, lentils, peas, and soybean, raw and vapor cooked) were compared to adult house crickets (Acheta domesticus) and mealworm larvae (Tenebrio molitor) that had been autoclaved, oven cooked (150 and 200°C), or used raw. Ingredients were run in an in vitro model of the pig gastrointestinal tract combining enzymes to simulate digestion in the stomach and the small intestine and subsequent fermentation by fecal microbes to simulate hindgut fermentation. In vitro crude protein disappearance (IVCPD) of insects decreased with oven cooking at 150°C or autoclaving (P < 0.05) while that of plants was unaffected (P > 0.05), except for soybean. IVCPD of raw mealworms (0.726) equaled that of the best plants (0.725 to 0.763) while crickets were less digestible (P < 0.01). Consequences on fermentation metabolites were lower propionate (P < 0.01) and branched-chain fatty acids (BCFA; P < 0.05) molar ratio in raw insects against oven-cooked or autoclaved insects. Both insect sources displayed greater BCFA (P < 0.01) and lower propionate (P < 0.01) than plants. Crickets produced 50% as much BCFA as mealworms (P < 0.01). In conclusion, feeding insect-sourced protein requires a careful choice of the species as well as the thermal treatment to avoid possible detrimental consequences on digestibility and intestinal health in pigs. [less ▲]

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See detailThe cultivable surface microbiota of the brown alga Ascophyllum nodosum is enriched in macroalgal-polysaccharide-degrading bacteria
Martin, Marjolaine ULiege; Barbeyron, Tristan; Martin, Renée ULiege et al

in Frontiers in Microbiology (2015), 6(december),

Bacteria degrading algal polysaccharides are key players in the global carbon cycle and in algal biomass recycling. Yet the water column, which has been studied largely by metagenomic approaches, is poor ... [more ▼]

Bacteria degrading algal polysaccharides are key players in the global carbon cycle and in algal biomass recycling. Yet the water column, which has been studied largely by metagenomic approaches, is poor in such bacteria and their algal-polysaccharide-degrading enzymes. Even more surprisingly, the few published studies on seaweed-associated microbiomes have revealed low abundances of such bacteria and their specific enzymes. However, as macroalgal cell-wall polysaccharides do not accumulate in nature, these bacteria and their unique polysaccharidases must not be that uncommon. We, therefore, looked at the polysaccharide-degrading activity of the cultivable bacterial subpopulation associated with Ascophyllum nodosum. From A. nodosum triplicates, 324 bacteria were isolated and taxonomically identified. Out of these isolates, 78 (∼25%) were found to act on at least one tested algal polysaccharide (agar, ι- or κ-carrageenan, or alginate). The isolates “active” on algal-polysaccharides belong to 11 genera: Cellulophaga, Maribacter, Algibacter, and Zobellia in the class Flavobacteriia (41) and Pseudoalteromonas, Vibrio, Cobetia, Shewanella, Colwellia, Marinomonas, and Paraglaceciola in the class Gammaproteobacteria (37). A major part represents likely novel species. Different proportions of bacterial phyla and classes were observed between the isolated cultivable subpopulation and the total microbial community previously identified on other brown algae. Here, Bacteroidetes and Gammaproteobacteria were found to be the most abundant and some phyla (as Planctomycetes and Cyanobacteria) frequently encountered on brown algae weren’t identified. At a lower taxonomic level, twelve genera, well-known to be associated with algae (with the exception for Colwellia), were consistently found on all three A. nosodum samples. Even more interesting, 9 of the 11 above mentioned genera containing polysaccharolytic isolates were predominant in this common core. The cultivable fraction of the bacterial community associated with A. nodosum is, thus, significantly enriched in macroalgal-polysaccharide-degrading bacteria and these bacteria seem important for the seaweed holobiont even though they are under-represented in alga-associated microbiome studies. [less ▲]

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See detailBiotechnological potential of the microflora associated with the brown alga Ascophyllum nodosum
Martin, Marjolaine ULiege; Martin, Renée ULiege; Barbeyron, Tristan et al

Conference (2015, August 18)

Bacteria associated with algae are underexplored despite their huge biodiversity and the fact that they differ markedly from those living freely in seawater. These bacterial communities are known to ... [more ▼]

Bacteria associated with algae are underexplored despite their huge biodiversity and the fact that they differ markedly from those living freely in seawater. These bacterial communities are known to represent great potential for the production of diverse bioactive compounds, such as specific glycoside hydrolases, as they interact in multiple complex ways with their host. Furthermore, enzymes from marine bacteria have original properties, like cold-adapted, halotolerant and highly stable, which are constantly searched out by bio-industries. The aim of our study was to identify bacteria, associated with the brown alga Ascophyllum nodosum, showing diverse polysaccharolytic activities. To isolate cultivable microorganisms, algal thalli of Ascophyllum nodosum were swabbed with sterile cotton tips and marine agar plates were inoculated. Three-hundred isolated bacteria were screened for agarase, kappa- and iota-carrageenase, and sulfatase activities on specific marine media. Thirty-two bacteria with polysaccharolytic activities were isolated and a part of their 16S rDNA (8F-1492R) were amplified and sequenced. Twenty-seven were classified as Flavobacteriia and five as Gammaproteobacteria. Putative new strains and species of Zobellia, Maribacter, Cellulophaga, Shewanella, Glaciecola, Pseudoalteromonas and Colwellia were identified by phylogenetic analysis. All those genera are well-known to colonize algal surface but only some of them are famous to degrade algal polysaccharides (Zobellia, Maribacter, Cellulophaga, and Pseudoalteromonas). However, all those novel bacterial strains/species showed multiple and diverse enzymatic activities (agarase, iota-and kappa-carrageenase, cellulase, beta-glucosidase, sulfatase and/or amylase activities). Genomics libraries with their DNA were constructed in Escherichia coli and Bacillus subtilis and are screened to identify the genes coding for the observed enzymatic activities. Those novel glycoside hydrolases from unknown marine bacteria should have original and innovative properties with great biotechnological potential. [less ▲]

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See detailIn vitro evaluation of fermentation characteristics of two types of insects, as potential novel protein feeds for pigs
POELAERT, Christine ULiege; Beckers, Yves ULiege; Despret, Xavier et al

in 13th International Symposium on Digestive Physiology of Pigs (2015, May)

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See detailNew lipolytic enzymes identified by screening two metagenomic libraries derived from the soil of a winter wheat field
Stroobants, Aurore ULiege; Martin, Renée ULiege; Roosens, Lyse et al

in Biotechnologie, Agronomie, Société et Environnement (2015), 19(2), 125-131

Lipolytic enzymes are widely distributed and fulfil important physiological functions in the microorganisms inhabiting diverse environments. Soils are rich, diversified environments containing microbial ... [more ▼]

Lipolytic enzymes are widely distributed and fulfil important physiological functions in the microorganisms inhabiting diverse environments. Soils are rich, diversified environments containing microbial communities that remain largely unknown. [less ▲]

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See detailIdentification and Characterization of a Halotolerant, Cold-Active Marine Endo-β-1,4-Glucanase by Using Functional Metagenomics of Seaweed-Associated Microbiota
Martin, Marjolaine ULiege; Biver, Sophie ULiege; Steels, Sébastien ULiege et al

in Applied and Environmental Microbiology (2014), 80(16), 4958-4967

A metagenomic library was constructed from microorganisms associated with the brown alga Ascophyllum nodosum. Functional screening of this library revealed 13 novel putative esterase loci and two ... [more ▼]

A metagenomic library was constructed from microorganisms associated with the brown alga Ascophyllum nodosum. Functional screening of this library revealed 13 novel putative esterase loci and two glycoside hydrolase loci. Sequence and gene cluster analysis showed the wide diversity of the identified enzymes and gave an idea of the microbial populations present during the sample collection period. Lastly, an endo-β-1,4-glucanase having less than 50% identity to sequences of known cellulases was purified and partially characterized, showing activity at low temperature and after prolonged incubation in concentrated salt solutions. [less ▲]

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See detailNew carbohydrate-active enzymes identified by screening two metagenomic libraries derived from the soil of a winter wheat field
Stroobants, Aurore ULiege; Portetelle, Daniel ULiege; Vandenbol, Micheline ULiege

in Journal of Applied Microbiology (2014)

Aims Soils are rich, diversified environments where β-glucosidases abound because of their importance in organic matter degradation. The aim of this work was to discover new β-glucosidases by constructing ... [more ▼]

Aims Soils are rich, diversified environments where β-glucosidases abound because of their importance in organic matter degradation. The aim of this work was to discover new β-glucosidases by constructing two metagenomic DNA libraries from soil samples collected in winter and spring from a field of winter wheat. Methods and Results Both libraries were screened on esculin-supplemented medium so as to isolate candidates showing β-glucosidase activity. Candidate analysis revealed seven putative β-glycosidases and two putative glycosyltransferases, displaying 25 to 82% identity to known enzymes. The putative β-glycosidases belong to families GH1, GH3 and GH20 and the two putative glycosyltransferases, probably, to new families. In characterization tests performed on bacteria in suspension or spread on agar plates, some candidates appeared to hydrolyse several natural and synthetic substrates. These tests also highlighted interesting industrial characteristics, such as the activity of four β-glycosidases under alkaline conditions and the esculin-hydrolysing activity of a β-glucosidase candidate in the presence of glucose. Conclusions Seven putative β-glycosidases and two putative glycosyltransferases were found by functional screening of two metagenomic DNA libraries derived from agricultural soil. Significance and Impact of the Study This study has identified β-glycosidases and putative glycosyltransferases that have or may have interesting industrial characteristics. [less ▲]

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See detailDiversity of Bacterial Communities in a Profile of a Winter Wheat Field: Known and Unknown Members
Stroobants, Aurore ULiege; Degrune, Florine ULiege; Olivier, Claire et al

in Microbial Ecology (2014)

In soils, bacteria are very abundant and diverse. They are involved in various agro-ecosystem processes such as the nitrogen cycle, organic matter degradation, and soil formation. Yet, little is known ... [more ▼]

In soils, bacteria are very abundant and diverse. They are involved in various agro-ecosystem processes such as the nitrogen cycle, organic matter degradation, and soil formation. Yet, little is known about the distribution and composition of bacterial communities through the soil profile, particularly in agricultural soils, as most studies have focused only on topsoils or forest and grassland soils. In the present work, we have used bar-coded pyrosequencing analysis of the V3 region of the 16S rRNA gene to analyze bacterial diversity in a profile (depths 10, 25, and 45 cm) of a well-characterized field of winter wheat. Taxonomic assignment was carried out with the Ribosomal Database Project (RDP) Classifier program with three bootstrap scores: a main run at 0.80, a confirmation run at 0.99, and a run at 0 to gain information on the unknown bacteria. Our results show that biomass and bacterial quantity and diversity decreased greatly with depth. Depth also had an impact, in terms of relative sequence abundance, on 81 % of the most represented taxonomic ranks, notably the ranks Proteobacteria, Bacteroidetes, Actinobacteridae, and Acidobacteria. Bacterial community composition differed more strongly between the topsoil (10 and 25 cm) and subsoil (45 cm) than between levels in the topsoil, mainly because of shifts in the carbon, nitrogen, and potassium contents. The subsoil also contained more unknown bacteria, 53.96 % on the average, than did the topsoil, with 42.06 % at 10 cm and 45.59 % at 25 cm. Most of these unknown bacteria seem to belong to Deltaproteobacteria, Actinobacteria, Rhizobiales, and Acidobacteria. [less ▲]

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See detailScreening of two agricultural genomic DNA libraries to seek new glycoside hydrolases
Stroobants, Aurore ULiege; Portetelle, Daniel ULiege; Vandenbol, Micheline ULiege

Poster (2014, February 07)

Soils are very rich environments where the diversity of microorganisms is very high. These microorganisms play an important role in the degradation of organic matter with enzymes able to degrade it. This ... [more ▼]

Soils are very rich environments where the diversity of microorganisms is very high. These microorganisms play an important role in the degradation of organic matter with enzymes able to degrade it. This work aims to discover, by functional screening, new microbial glycoside hydrolases from soils collected in winter and spring in a winter wheat crop. The genomic DNA was extracted from both soils to construct two libraries in Escherichia coli. These libraries were then screened for beta-glucosidase activities on 2YT agar media containing 0.5% esculin and 0.1% ammonium iron (III) citrate. At this time, about 250.000 clones from each library have been screened. Two beta-glucosidases have already been found in the winter library while five beta-glucosidases and two glycosyltransferases were identified in the spring library. Sequence analyses with the BLASTX program revealed putative enzymes showing between 25% and 72% sequence identity with known enzymes and belonging to three glycoside hydrolase families (GH1, GH3 and GH20) and to two probably new glycosyltransferase families. Biochemical characterisation of the candidates at several pH values and temperatures, and with four substrates, is in progress. [less ▲]

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See detailProjet Termitofuel: les termites et leurs symbiontes pour mieux valoriser la biomasse ligno-cellulosique
Bauwens, Julien ULiege; Tarayre, Cédric ULiege; Brasseur, Catherine ULiege et al

Poster (2013, October 19)

L’exploitation actuelle de la biomasse ligno-cellulosique, extrêmement abondante sur Terre, est liée à la production de déchets relativement peu valorisables. C’est pourquoi le projet TERMITOFUEL étudie ... [more ▼]

L’exploitation actuelle de la biomasse ligno-cellulosique, extrêmement abondante sur Terre, est liée à la production de déchets relativement peu valorisables. C’est pourquoi le projet TERMITOFUEL étudie la digestion du bois chez les termites, grâce à une approche pluridisciplinaire, en vue d’améliorer la production de bioéthanol de seconde génération. Au sein de l’Unité d’Entomologie Fonctionnelle et Evolutive, l’approche protéomique est employée afin d’identifier les micro-organismes présents dans le tube digestif des termites et caractériser les activités enzymatiques. [less ▲]

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See detailFunctional screening of a winter and a spring genomic DNA libraries obtained from soils in a winter wheat crop
Stroobants, Aurore ULiege; Portetelle, Daniel ULiege; Vandenbol, Micheline ULiege

Poster (2013, June 10)

Soils are very rich environments where the diversity of microorganisms is very high. These microorganisms play important role in the degradation of organic matter with enzymes able to degrade it. The aim ... [more ▼]

Soils are very rich environments where the diversity of microorganisms is very high. These microorganisms play important role in the degradation of organic matter with enzymes able to degrade it. The aim of this work is to discover by functional screening new enzymatic activities of microorganisms from soils collected in winter and spring in a winter wheat crop. The genomic DNA was extracted from both soils to construct two libraries in Escherichia coli. These libraries were then screened for several enzymes such as lipase, beta-glucosidase, cellulase, α-amylase,… At this time, 2 beta-glucosidases and 3 lipases have already been found in the winter library and 3 beta-glucosidases and 1 lipase in the spring library. Sequence analyses with the BLASTX program revealed that two beta-glucosidases have less than 65% of sequence identity with known beta-glucosidases, one have 64% of identity with a known beta-galactosidase and one have 59% of identity with a glycoside hydrolase. The fifth seems to be a phosphorylase kinase (54% identity) which have a glucoamylase domain responsible for the activity. This ORF is interrupted by a transposase. Three of the four lipases have less than 60% of sequence identity with known lipases/esterases. The fourth show 55% of identity with a known beta-lactamase. [less ▲]

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See detailCharacterization of new bacterial glycoside hydrolases isolated from agricultural soils using a functional metagenomic approach
Biver, Sophie ULiege; Dubois, Benjamin; Stroobants, Aurore ULiege et al

Poster (2013, June 10)

Microorganisms play key roles in soil ecosystem functioning, notably through their ability to degrade plant cell wall polymers. For this, bacteria and fungi produce various enzymes such as cellulases ... [more ▼]

Microorganisms play key roles in soil ecosystem functioning, notably through their ability to degrade plant cell wall polymers. For this, bacteria and fungi produce various enzymes such as cellulases, xylanases, glucosidases, esterases or laccases. Finding new enzymes hydrolyzing cellulose, hemicellulose or lignin is not only interesting for a better understanding of the roles of the soil microflora still largely unknown but these enzymes are also useful for various biotechnological applications such as the production of renewable energy from lignocellulosic material. So here, we used a functional metagenomic approach to isolate new bacterial β-glucosidases, which were then biochemically characterized. The new enzymes were identified by functional analysis of agricultural-soil metagenomic libraries hosted in Escherichia coli and screened on medium containing esculin. After sequence analysis and preliminary estimation of the activity of the new β-glucosidases using p-nitrophenol derivatives on intact bacterial cells, the coding sequences of three of them were cloned into a bacterial expression vector so as to overproduce and purify them by affinity chromatography. The chosen enzymes show only 52-64% sequence identity to known family 3 (GH3) or 1 (GH1) glycoside hydrolases of different phyla (Actinobacteria, Acidobacteria and Proteobacteria). Analysis of the E. coli cells expressing each of them revealed that both GH1 proteins (ASEsc9 and ASEsc10) are thermophilic enzymes more active at mildly acidic to neutral pH while the GH3 enzyme (ASEsc6) is an alkaline, mesophilic, β-glucosidase also displaying xylosidase activity. Their coding sequences have been cloned in fusion with a carboxy-terminal His-tag and placed under the control of the IPTG-inducible promoter of the pET-30b vector. The proteins will be overproduced and purified for further characterization. [less ▲]

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See detailStudy of bacterial diversity in the topsoil and below the hardpan in an agricultural soil by metagenomics following by two analysis pipelines
Stroobants, Aurore ULiege; Lambert, Christrophe; Degrune, Florine ULiege et al

Poster (2013, June 10)

On earth, Bacteria are ubiquitous and even present in extreme environments (pH, temperature,…). In soils in particular, bacteria are very abundant (up to 109 cells per gram of soil) but still poorly ... [more ▼]

On earth, Bacteria are ubiquitous and even present in extreme environments (pH, temperature,…). In soils in particular, bacteria are very abundant (up to 109 cells per gram of soil) but still poorly characterized. Thus, it is of paramount importance to use relevant study and analysis procedures to ensure that the results obtained closely reflect the real-life conditions. In the present work, we analyze the bacterial diversity in the topsoil and below the hardpan in an agricultural soil using the metagenomics approach, with the Ion Torrent PGM sequencer. The soil samples was collected at three depths : 10 cm (topsoil), 25 cm (topsoil above the hardpan) and 45 cm (below the hardpan), in a tilled and a no tilled plot. The taxonomic analysis of the reads obtained are carried out according to two different procedures with the RDP classifier program and with a confidence score threshold of 0 and 0.99. The 0 threshold is used to assign a species to all reads, each read being therefore assigned to its most closest known species. The threshold of 0.99 enables us to focus on reads being assigned to a species with a high degree of confidence. In this case, each read is assigned to the most specific rank having a confidence score higher than 0.99. The bacterial diversity was then compared between the different conditions. Results obtained demonstrate that the bacterial communities were not the same in the two horizons. For example, some classes of Acidobacteria were up to 11 fold more numerous in topsoil while others was until 12 fold more represented below the hardpan. The biomass and the bacterial diversity (Shannon index) were also greatly different between the two depths. [less ▲]

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