In Vitro Characterization of the Impact of Different Substrates on Metabolite Production,Energy Extraction and Composition of Gut Microbiota from Lean and Obese Subjects

The aim of this study was to investigate the effect of galacto-oligosaccharides, lactulose, apple fiber and sugar beet pectin on the composition and activity of human colonic microbiota of lean and obese healthy subjects using an in vitro model of the proximal colon: TIM-2. Substrate fermentation was assessed by measuring the production of short-chain and branched-chain fatty acids, lactate and ammonia and by studying the composition of the bacterial communities over time. The results suggest that energy harvest (in terms of metabolites) of lean and obese microbiotas is different and may depend on the fermentable substrate. For galacto-oligosaccharides and lactulose, the cumulative amount of short-chain fatty acids plus lactate produced in TIM-2 was lower in the fermentation experiments with the lean microbiota (123 and 155 mmol, respectively) compared to the obese (162 and 173 mmol, respectively). This was reversed for the pectin and the fiber. The absolute amount produced of short-chain fatty acids including lactate was higher after 72 h in the fermentation experiments with apple fiber-L (108 mmol) than with apple fiber-O (92 mmol). Sugar beet-L was also higher (130 mmol) compared to sugar beet-O (103 mmol). Galacto-oligosaccharides and lactulose boosted the balance of health-promoting over toxic metabolites produced by the microbiota from obese subjects. Firmicutes were more predominant in the inoculum prepared from feces of obese subjects compared to lean subjects. The average abundance at time zero was 92% and 74%, respectively. On the other hand, Bacteroidetes were more dominant in the microbiota prepared with homogenates from lean subjects with an average abundance of 22% compared with the microbiota prepared with homogenates from obese subjects (3.6%). This study brings evidence that different fermentable carbohydrates are fermented differently by lean and obese microbiotas, which contributes to the understanding of the role of diet and the microbiota in tackling obesity.     

Species Traits as Practical Tools for Ecological Restoration of Marly Eroded Lands

Plant functional traits are increasingly used in restoration ecology because they have the potential to guide restoration practices at a broad scale. This article presents a trait‐based multi‐criteria framework to evaluate and predict the performance of 17 plant seedlings to improve ecological restoration of marly eroded areas in the French Southern Alps. The suitability of these species to limit soil erosion was assessed by studying both their response to erosive forces and their effect on erosion dynamics. We assumed that species efficiency could be explained and predicted from plant traits and we looked for trait‐performance relationships. Our results showed that root slenderness ratio, the percentage of fine roots and root system topology, were the three root morphology traits best describing anchorage strength. Root system characterized by a long and thin tap root and many fine lateral ramifications would be the best to resist concentrated runoff. Species response to burial mainly depended on growth form and morphological flexibility. The abilities of species in reinforcing the soil and reducing erosion rates were negatively correlated to root diameter and positively to the percentage of fine roots. Moreover, root system density and root tensile strength also influenced root reinforcement. Finally, the ability to trap sediment was positively correlated to leaf area and canopy density. Species were then scored and classified in four clusters according to their global performance. This method allows identifying species that possess both response and effect traits related to the goal of preventing erosion during ecological restoration.     

A METHOD FOR THE CRYOCONSERVATION OF DUNALIELLA SALINA (CHLOROPHYCEAE): EFFECT OF GLYCEROL AND COLD ADAPTATION1

The unicellular green alga Dunaliella salina Teod. was frozen according to the following procedure: 3 days cold adaptation at 4°C, addition of 3.5 M glycerol as a cryoprotectant, slow cooling to –40°C, immersion in liquid nitrogen, and rapid thawing. The survival rate was higher when cells were grown, before freezing, in the presence of 2 M NaCl instead of 1 M NaCl (78 and 48% survival, respectively). This difference is probably due to the intracellular amount of glycerol, which increases with external NaCl concentration and, therefore, may enhance cell protection. Although cells grown in 4 M NaCl accumulated a large amount of glycerol in response to osmotic stress, they did not withstand freezing. The use of cryoprotectant was absolutely necessary for the cells to recover from storage at –196°C. Glycerol was used because it is naturally produced by Dunaliella salina and therefore is not toxic. Provided it was added slowly to avoid osmotic shock, 3.5 M glycerol gave better results than 1M glycerol (48 and 18% survival, respectively). Cold adaptation in the dark increased postthaw viability. Cells grown in 1 M or 2 M NaCl had a survival rate of 48 and 78%, respectively, when cold-adapted, against 10 and 42% when not cold-adapted. This adaptation could be due to the synthesis, at low temperature, of specific proteins because two bands (28–29 kDa) appeared when electrophoretically separated proteins from cold-adapted cells and control cells were compared. Also, it could be due to the degradation of starch that occurs in the dark and leads to glycerol accumulation. Our procedure has never been used to cryopreserve microalgae and could enhance reported survival rates.     

Quenching autofluorescence in the alimentary canal tissues of Bactericera cockerelli (Hemiptera: Triozidae) for immunofluorescence labeling

Immunofluorescence has been widely used to localize microbes or specific molecules in insect tissues or cells. However, significant autofluorescence is frequently observed in tissues which can interfere with the fluorescent identification of target antigens, leading to inaccurate or even false positive fluorescent labeling. The alimentary canal of the potato psyllid, Bactericera cockerelli ?ulc, exhibits intense autofluorescence, hindering the application of immunolocalization for the detection and localization of the economically important pathogen transmitted by this insect, “Candidatus Liberibacter solanacearum” (Lso). In the present study, we tested the use of irradiation, hydrogen peroxide (H₂O₂) and Sudan black B (SBB) treatments to reduce the autofluorescence in the B. cockerelli alimentary canal tissues. Furthermore, we assessed the compatibility of the above‐mentioned treatments with Lso immunolocalization and actin staining using phalloidin. Our results showed that the autofluorescence in the alimentary canal was reduced by irradiation, H₂O₂, or SBB treatments. The compatibility assays indicated that irradiation and H₂O₂ treatment both greatly reduced the fluorescent signal associated with Lso and actin. However, the SBB incubation preserved those target signals, while efficiently eliminating autofluorescence in the psyllid alimentary canal. Therefore, herein we propose a robust method for reducing the autofluorescence in the B. cockerelli alimentary canal with SBB treatment, which may improve the use of immunofluorescence labeling in this organism. This method may also have a wide range of uses by reducing the autofluorescence in other arthropod species.     

Crystallographic structure of metal-free concanavalin A at 2.5 Å resolution

The three-dimensional structure of demetallized concanavalin A has been determined at 2.5 Å resolution and refined to a crystallographic R-factor of 18%. The lectin activity of concanavalin A requires the binding of both a transition metal ion, generally Mn²⁺, and a Ca²⁺ ion in two neighboring sites in close proximity to the carbohydrate binding site. Large structural differences between the native and the metal-free lectin are observed in the metal-binding region and consequently for the residues involved in the specific binding of saccharides. The demetallization invokes a series of conformational changes in the protein backbone, apparently initiated mainly by the loss of the calcium ion. Most of the Mn²⁺ ligands retain their position, but the Ca²⁺ binding site is destroyed. The Ala207-Asp208 peptide bond, in the β-strand neighboring the metal-binding sites, undergoes a cis to trans isomerization. The cis conformation for this bond is a highly conserved feature among the leguminous lectins and is critically maintained by the Ca²⁺ ion in metal-bound concanavalin A. A further and major change adjacent to the isomerized bond is an expansion of the loop containing the monosaccharide ligand residues Leu99 and Tyr100. The dispersion of the ligand residues for the monosaccharide binding site (Asn14, Agr228, Asp208, Leu99, and Tyr100) in metalfree concanavalin A abolishes the lectin's ability to bind saccharides. Since the quaternary structure of legume lectins is essential to their biological role, the tetramer formation was analyzed. In the crystal (pH 5), the metal-free concanavalin A dimers associate into a tetramer that is similar to the native one, but with a drastically reduced number of inter-dimer interactions. This explains the tetramer dissociation into dimers below pH values of 6.5. © 1995 Wiley-Liss, Inc.     

Genotypic proportions in hybrid zones

We study a hybrid zone between two populations of a diploid organism. The populations differ at one locus. Homozygotes have equal fitnesses and the heterozygote fitness is reduced by + ( is the birth rate deviation and is the death rate deviation). The populations extend along a one dimensional continuous habitat, and migration occurs by diffusion of individuals. The model is formulated as a set of simple continuous time demographic models without age structure for the three genotypes, and the system is transformed into three new variables, the total population size N, the gene frequency p, and the deviation from Hardy-Weinberg proportions F. The gene frequency in a steady state cline always follows a hyperbolic tangent closely. Analysis of the asymptotic behavior of the cline far from the hybrid zone suggests a qualitative prediction of the shape of N, p and F over the zone. For weak selection the shape is determined by a central steepness of , as observed by Bazykin in 1969, where D is the diffusion coefficient. For strong selection the cline is less steep than the Bazykin cline, and the form is dominated by the migration process. The steepness at the center of the cline is close to where b is the birth rate of homozygotes.Supported in part by grant 11-9639-1 from the Danish Natural Science Research Council and grant 5.18.2023 from the Danish Biotechnological Research and Development Programme     

Incubation of innovative methanogenic communities to seed anaerobic digesters

Applied Microbiology and Biotechnology - The methanogenic communities in alternative inocula and their potential to increase CH4 production in mesophilic and psychrophilic dairy manure-based...     

A two-locus mutation—Selection model and some of its evolutionary implications

The deterministic properties of a two-locus model with mutation and selection have been investigated. The mutation process is unidirectional, and the model is so constructed that the genetic variation at one locus is selectively neutral in the absence of a mutant allele at the other locus. All genotypes with three or four mutant alleles are deleterious, while the double heterozygotes may have the same fitness as the standard genotype. If one of the mutant alleles becomes fixed in the population, then the other locus will show a regular one-locus mutation-selection balance. Such a boundary equilibrium may be unstable or stable in the full two-locus setting. In the symmetric case, which is analyzed in details, the population will either go to one of the two boundary equilibria, or to a fully polymorphic equilibrium at which both the mutant alleles are rare. The origin of reproductive separation between two populations via the fixation of complementary deleterious mutants at different loci, and the fixation of nonfunctional alleles at duplicated loci, are two biological processes which both can be studied with the present model. In the last part of the paper we show how the results from the deterministic analysis can be used to predict how different factors will influence the rates of evolution in these systems.     

Geographical Patterns of Four Polymorphisms in ZOARCES VIVIPARUS as Evidence of Selection

Two unlinked polymorphisms of a marine teleost, one in hemoglobin and one in esterase, show parallel clines through Danish waters, while two unlinked phosphoglucosemutase polymorphisms maintain constant gene frequencies. It is argued that the clinal and the constant polymorphisms cannot be accounted for simultaneously by random genetic drift of selectively neutral genes. It is therefore concluded that selection is responsible for at least one of the two classes of geographical patterns, be it the clinal, the constant, or both.     

Molluscicidal activity of the Puerto Rican Weed,Solatium nodiflorum,against snail hosts ofFasciola hepatica

After screening 200 species of Puerto Rican plants, Solanum nodiflorumwas selected for comprehensive laboratory evaluation as a source of natural molluscicide. All parts of this plant were found toxic to the snails Fossaria (Lymnaea) cubensis, Lymnaea columella, Biomphalaria glabrata,and Physa cubensis.No toxicity was found against Marisa cornuarietisand Tarebia granifera. The toxicant was not affected by pH and dry storage. There was a direct correlation between temperature and molluscicidal activity. The plant is easy to find locally and could be developed into cheap, safe means for controlling both Schistosomaand Fasciolasnail hosts in Puerto Rico.