Placenta-like structure of the aphid endoparasitic wasp Aphidius ervi: a strategy of optimal resources acquisition

Aphidius ervi (Hymenoptera: Braconidae) is an entomophagous parasitoid known to be an effective parasitoid of several aphid species of economic importance. A reduction of its production cost during mass rearing for inundative release is needed to improve its use in biological control of pests. In these contexts, a careful analysis of its entire development phases within its host is needed. This paper shows that this parasitoid has some characteristics in its embryological development rather complex and different from most other reported insects, which can be phylogenetically very close. First, its yolkless egg allows a high fecundity of the female but force them to hatch from the egg shell rapidly to the host hemocoel. An early cellularisation allowing a rapid differentiation of a serosa membrane seems to confirm this hypothesis. The serosa wraps the developing embryo until the first instar larva stage and invades the host tissues by microvilli projections and form a placenta like structure able to divert host resources and allowing nutrition and respiration of embryo. Such interspecific invasion, at the cellular level, recalls mammal's trophoblasts that anchors maternal uterine wall and underlines the high adaptation of A. ervi to develop in the host body.     

Stochastic models to study the impact of mixing on a fed-batch culture of Saccharomyces cerevisiae

The mechanisms of interaction between microorganisms and their environment in a stirred bioreactor can be modeled by a stochastic approach. The procedure comprises two submodels: a classical stochastic model for the microbial cell circulation and a Markov chain model for the concentration gradient calculus. The advantage lies in the fact that the core of each submodel, i.e., the transition matrix (which contains the probabilities to shift from a perfectly mixed compartment to another in the bioreactor representation), is identical for the two cases. That means that both the particle circulation and fluid mixing process can be analyzed by use of the same modeling basis. This assumption has been validated by performing inert tracer (NaCl) and stained yeast cells dispersion experiments that have shown good agreement with simulation results. The stochastic model has been used to define a characteristic concentration profile experienced by the microorganisms during a fermentation test performed in a scale-down reactor. The concentration profiles obtained in this way can explain the scale-down effect in the case of a Saccharomyces cerevisiae fed-batch process. The simulation results are analyzed in order to give some explanations about the effect of the substrate fluctuation dynamics on S. cerevisiae.     

Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis

In this work, the potential of Bacillus subtilis strain M4 at protecting plants against fungal diseases was demonstrated in different pathosystems. We provide evidence for the role of secreted lipopeptides, and more particularly of fengycins, in the protective effect afforded by the strain against damping-off of bean seedlings caused by Pythium ultimum and against gray mold of apple in post-harvest disease. This role was demonstrated by the strong biocontrol activity of lipopeptide-enriched extracts and through the detection of inhibitory quantities of fengycins in infected tissues. Beside such a direct antagonism of the pathogen, we show that root pre-inoculation with M4 enabled the host plant to react more efficiently to subsequent pathogen infection on leaves. Fengycins could also be involved in this systemic resistance-eliciting effect of strain M4, as these molecules may induce the synthesis of plant phenolics involved in or derived from the defense-related phenylpropanoid metabolism. Much remains to be discovered about the mechanisms by which Bacillus spp suppress disease. Through this study on strain M4, we reinforce the interest in B. subtilis as a pathogen antagonist and plant defense-inducing agent. The secretion of cyclic fengycin-type lipopeptides may be tightly related to the expression of these two biocontrol traits.     

Influence of compressed fluids treatment on the activity of Yarrowia lipolytica lipase

This work investigated the influence of temperature, pressure, exposure times and depressurization rate on the activity of a non-commercial immobilized lipase from Yarrowia lipolytica (YLL) submitted to compressed carbon dioxide, propane and n-butane. A high-pressure cell was employed in the experiments, in the pressure range of 10–280 bar, varying the temperature from 35 to 75 °C, exposure times from 1 to 6 h, and adopting distinct decompression rates. Results showed that significant activity losses were obtained when the treatment was conducted in carbon dioxide, while negligible losses were observed in both propane and n-butane. For the treatment with carbon dioxide, within the range studied, the decompression rate affected positively enzyme activity, while the exposure time and temperature presented an opposite effect on the non-commercial immobilized lipase from Y. lipolytica (YLL). Additionally, the performance of two commercial immobilized lipases (Lipozyme IM and Novozym 435) and the immobilized YLL in the three solvents was compared. Immobilized YLL has shown to be more suitable than Lipozyme IM for enzyme-catalyzed reactions using compressed propane and n-butane as solvents, but with inferior performance compared to Novozym 435 treated in these solvents.     

Proteolytic breakdown of gliadin by <Emphasis Type="Italic">Enterococcus faecalis</Emphasis> isolated from Tunisian fermented dough

The aim of this work was to select strains with proteolytic activity on wheat gliadin, among lactic acid bacteria, previously isolated from Tunisian fermented wheat dough. Hydrolysis of gliadin, as sole nitrogen source, in an agar medium was visualized by a clear zone surrounding colonies. The increase in absorbance due to gliadin breakdown was measured spectrophotometrically using O-phthaldialdehyde (OPA) on Gliadin Glucose Broth medium. Fermented liquid dough inoculated with individual selected Enterococcus faecalis, showed a decrease of the gliadin concentration from 45 g/kg to 18 g/kg determined by sandwich ELISA test (R-7001). Only the enterococci strains show an hydrolysis of gliadin proteins. Strains showing proteolytic activity are gaining more and more importance in cereal based fermented foods and may help to reduce gliadin involved in coeliac disease.     

The source of fermentable carbohydrates influences the in vitro protein synthesis by colonic bacteria isolated from pigs

Two in vitro experiments were carried out to quantify the incorporation of nitrogen (N) by pig colonic bacteria during the fermentation of dietary fibre, including non-starch polysaccharides and resistant starch. In the first experiment, five purified carbohydrates were used: starch (S), cellulose (C), inulin (I), pectin (P) and xylan (X). In the second experiment, three pepsin-pancreatin hydrolysed ingredients were investigated: potato, sugar-beet pulp and wheat bran. The substrates were incubated in an inoculum, prepared from fresh faeces of sows and a buffer solution providing 15N-labelled NH4Cl. Gas production was monitored. Bacterial N incorporation (BNI) was estimated by measuring the incorporation of 15N in the solid residue at half-time to asymptotic gas production (T/2). The remaining substrate was analysed for sugar content. Short-chain fatty acids (SCFA) were determined in the liquid phase. In the first experiment, the fermentation kinetics differed between the substrates. P, S and I showed higher rates of degradation (P < 0.001), while X and C showed a longer lag time and T/2. The sugar disappearance reached 0.91, 0.90, 0.81, 0.56 and 0.46, respectively, for P, I, S, C and X. Among them, S and I fixed more N per gram substrate (P < 0.05) than C, X and P (22.9 and 23.2 mg fixed N per gram fermented substrate v. 11.3, 12.3 and 9.8, respectively). Production of SCFA was the highest for the substrates with low N fixation: 562 and 565 mg/g fermented substrate for X and C v. 290 to 451 for P, I and S (P < 0.01). In the second experiment, potato and sugar-beet pulp fermented more rapidly than wheat bran (P < 0.001). Substrate disappearance at T/2 varied from 0.17 to 0.50. BNI were 18.3, 17.0 and 10.2 fixed N per gram fermented substrate, for sugar-beet pulp, potato and wheat bran, respectively, but were not statistically different. SCFA productions were the highest with wheat bran (913 mg/g fermented substrate) followed by sugar-beet pulp (641) and potato (556) (P < 0.05). The differences in N uptake by intestinal bacteria are linked to the partitioning of the substrate energy content between bacterial growth and SCFA production. This partitioning varies according to the rate of fermentation and the chemical composition of the substrate, as shown by the regression equation linking BNI to T/2 and SCFA (r2 = 0.91, P < 0.01) and the correlation between BNI and insoluble dietary fibre (r = -0.77, P < 0.05) when pectin was discarded from the database.     

Bioreactor hydrodynamic effect on Escherichia coli physiology: experimental results and stochastic simulations

A microorganism circulating in a bioreactor can be submitted to hydrodynamic conditions inducing a significant effect on its physiology. The mixing time exhibited by the stirred bioreactor and the circulation of microorganisms are both involved in this reacting system. The mixing component determines the intensity of the concentration gradient and the circulation component determines the way in which the microorganism is exposed to this gradient. These two components linked to the experimental evaluation of microbial physiology can be analysed by a structured stochastic model in the case of a partitioned or “scale-down” reactor (SDR). A stochastic model indeed enables to simulate the mixing process as well as the circulation of microorganisms in SDRs. The superimposition of mixing and circulation processes determines the concentration profile experienced by a microorganism in the reactor. In the present case, the glucose concentration experienced by Escherichia coli has been modelled during a fed-batch culture. In this context, the use of a stochastic hydrodynamic model has permitted to point out an interesting feed pulse retardant effect in the SDRs. Nevertheless, the metabolic response of E. coli is not easy to interpret because of the possible simultaneous developments of overflow metabolism and mixed acid fermentation induced by the strong glucose concentration in the reactor.     

Fertilizer nitrogen budgets of 15N-labelled sugarbeet (Beta vulgaris) tops and Na 15NO3 dressings split-applied to winter wheat (Triticum aestivum) in microplots on a loam soil

The fate of N from sugarbeet (Beta vulgaris L.) tops returned to the soil (50 T ha^-1) in autumn 1986 before sowing winter wheat (Triticum aestivum L.), and from NaNO₃ split-applied in 3 equal dressings (at tillering, stem elongation and flag leaf stages) was studied using isotopically labelled ¹⁵N in open stainless-steel cylinders pressed into the soil.At harvest, the percentage utilization (PU) of N from sugarbeet was very low (6.66%) and negatively influenced by fertilizer N (5.59%), while that of fertilizer N was rather high (69.64%) and unchanged by addition of tops. Residual N in soil represented 25.9% of the amount applied in tops and ranged from 33% for the tillering application to 21% for the flag leaf application. N losses (mainly denitrification) from sugar beet tops amounted to 67% and were very low for mineral fertilizer (less than 5%).     

Improvement of lipase production from Yarrowia lipolytica

Extracellular lipase production by Yarrowia lipolytica was increased by mutant selection from 28 U/ml to 1000 U/ml. This activity was also reached in a 500 l bioreactor. The properties of the mutant lipase were the same of those of the wild type: M 38 kDa, optimum pH 7 and optimum temperature 37¡C.