Uncertainties in the adaptation of alpine pastures to climate change based on remote sensing products and modelling.

Over the last century, the management of pastoral systems has undergone major changes to meet the livelihood needs of alpine communities. Faced with the changes induced by recent global warming, the ecological status of many pastoral systems has seriously deteriorated in the western alpine region. We assessed changes in pasture dynamics by integrating information from remote-sensing products and two process-based models, i.e. the grassland-specific, biogeochemical growth model PaSim and the generic crop-growth model DayCent. Meteorological observations and satellite-derived Normalised Difference Vegetation Index (NDVI) trajectories of three pasture macro-types (high, medium and low productivity classes) in two study areas - Parc National des Écrins (PNE) in France and Parco Nazionale Gran Paradiso (PNGP) in Italy - were used as a basis for the model calibration work. The performance of the models was satisfactory in reproducing pasture production dynamics (R2 = 0.52 to 0.83). Projected changes in alpine pastures due to climate-change impacts and adaptation strategies indicate that: i) the length of the growing season is expected to increase between 15 and 40 days, resulting in changes in the timing and amount of biomass production, ii) summer water stress could limit pasture productivity; iii) earlier onset of grazing could enhance pasture productivity; iv) higher livestock densities could increase the rate of biomass regrowth, but major uncertainties in modelling processes need to be considered; and v) the carbon sequestration potential of pastures could decrease under limited water availability and warming.

Aflatoxin B1 adsorption/desorption dynamics in the presence of Lactobacillus rhamnosus RC007 in a gastrointestinal tract-simulated model.

AIMS: (i) To determine the aflatoxin B1 (AFB1 ) adsorption and desorption dynamics in the presence of Lactobacillus rhamnosus RC007 under simulated transit of AFB1 at each gastrointestinal tract (GIT-saliva, stomach and intestine) stage consecutively and then, separately, (ii) to study the ability of L. rhamnosus RC007 to biotransform AFB1 as a strategy that complements the adsorption process. METHODS AND RESULTS: The AFB1 adsorption and desorption assay simulating the GIT passage of AFB1 (93·89 ng g-1 ) in the presence of L. rhamnosus RC007 (108 CFU per ml) was conducted. Moreover, lactic acid production was determined. Results demonstrated that predominant environmental conditions in salivary solution induced a low AFB1 adsorption, while the transit through the gastric solution and intestinal solution allowed high percentages of adsorption and did not generate significant AFB1 desorption. CONCLUSIONS: The AFB1 adsorption and desorption dynamics in the presence of L. rhamnosus RC007 was favoured by gastric and intestinal environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The knowledge of the adsorption dynamics of AFB1 with a micro-organism of interest will allow predicting its behaviour at each stage of the GIT.

Improved robustness of an ethanologenic yeast strain through adaptive evolution in acetic acid is associated with its enzymatic antioxidant ability.

AIMS: To investigate multiple tolerance of Saccharomyces cerevisiae obtained through a laboratory strategy of adaptive evolution in acetic acid, its relation with enzymatic ROS detoxification and bioethanol 2G production. METHODS AND RESULTS: After adaptive evolution in acetic acid, a clone (Y8A) was selected for its tolerance to high acetic acid concentrations (13 g l-1 ) in batch cultures. Y8A was resistant to multiple stresses: osmotic, thermic, oxidative, saline, ethanol, organic acid, phenolic compounds and slow freeze-thawing cycles. Also, Y8A was able to maintain redox homeostasis under oxidative stress, whereas the isogenic parental strain (Y8) could not, indicating higher basal activity levels of antioxidative enzyme Catalase (CAT) and Gluthatione S-transferase (GST) in Y8A. Y8A reached higher bioethanol levels in a fermentation medium containing up to 8 g l-1 of acetic acid when compared to parental strain Y8. CONCLUSIONS: A multiple-stress-tolerant clone was obtained using adaptive evolution in acetic acid. Stress cross-tolerance could be explained by its enzymatic antioxidative capacity, namely CAT and GST. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrate that adaptive evolution used in S. cerevisiae was a useful strategy to obtain a yeast clone tolerant to multiple stresses. At the same time, our findings support the idea that tolerance to oxidative stress is the common basis for stress cotolerance, which is related to an increase in the specific enzymes CAT and GST but not in Superoxide dismutase, emphasizing the fact that detoxification of H2 O2 and not O2 ˙ is a key condition for multiple stress tolerance in S. cerevisiae.

Continuous and discontinuous dynamic unbinding transitions in drawn film flow.

When a plate is withdrawn from a liquid bath a coating layer is deposited whose thickness and homogeneity depend on the velocity and the wetting properties of the plate. Using a long-wave mesoscopic hydrodynamic description that incorporates wettability via a Derjaguin (disjoining) pressure we identify four qualitatively different dynamic transitions between microscopic and macroscopic coatings that are out-of-equilibrium equivalents of known equilibrium unbinding transitions. Namely, these are continuous and discontinuous dynamic wetting and emptying transitions. Several of their features have no equivalent at equilibrium.

Collapsed heteroclinic snaking near a heteroclinic chain in dragged meniscus problems.

A liquid film is studied that is deposited onto a flat plate that is inclined at a constant angle to the horizontal and is extracted from a liquid bath at a constant speed. We analyse steady-state solutions of a long-wave evolution equation for the film thickness. Using centre manifold theory, we first obtain an asymptotic expansion of solutions in the bath region. The presence of an additional temperature gradient along the plate that induces a Marangoni shear stress significantly changes these expansions and leads to the presence of logarithmic terms that are absent otherwise. Next, we numerically obtain steady solutions and analyse their behaviour as the plate velocity is changed. We observe that the bifurcation curve exhibits collapsed (or exponential) heteroclinic snaking when the plate inclination angle is above a certain critical value. Otherwise, the bifurcation curve is monotonic. The steady profiles along these curves are characterised by a foot-like structure that is formed close to the meniscus and is preceded by a thin precursor film further up the plate. The length of the foot increases along the bifurcation curve. Finally, we prove with a Shilnikov-type method that the snaking behaviour of the bifurcation curves is caused by the existence of an infinite number of heteroclinic orbits close to a heteroclinic chain that connects in an appropriate three-dimensional phase space the fixed point corresponding to the precursor film with the fixed point corresponding to the foot and then with the fixed point corresponding to the bath.

Statistical optimization of culture conditions for biomass production of probiotic gut-borne Saccharomyces cerevisiae strain able to reduce fumonisin B1.

AIM: To evaluate the ability of probiotic Saccharomyces cerevisiae RC016 strain to reduce fumonisin B(1) (FB(1)) in vitro and to optimize the culture conditions for the growth of the yeast employing surface response methodology. METHODS AND RESULTS: Using Plackett-Burman screening designs (PBSD) and central composite designs (CCD), an optimized culture medium containing (g l(-1)) fermentable sugars provided by sugar cane molasses (CMs), yeast extract (YE) and (NH(4))(2) HPO(4) (DAP) was formulated. The S. cerevisiae RC016 strain showed the greatest binding at all assayed FB1 concentration. The CMs, YE, DAP concentrations and incubation time influenced significantly the biomass of S. cerevisiae RC016. CONCLUSION: A combination of CMs 17%; YE 4·61 g l(-1) and incubation time 60 h was optimum for maximum biomass of S. cerevisiae RC016. SIGNIFICANCE AND IMPACT OF THE STUDY: The importance of this work lies in the search for live strains with both probiotic and fumonisin B1 decontamination properties that could be sustainably produced in a medium just containing cheap carbon, nitrogen and phosphorus sources and would be included in a novel product to animal feed.

Seasonal course of photosynthetic efficiency in Larix decidua Mill. in response to temperature and change in pigment composition during senescence.

This manuscript presents a study aimed at characterizing the seasonal course of photosynthetic capacity of an alpine deciduous conifer, European larch (Larix decidua Mill.), based on chlorophyll fluorescence measurements and photosynthetic pigment analysis. The study focused on the characterization of autumn senescence events which (contrary to bud-burst) are still scarcely investigated. The study was conducted on two natural European larch stands in the northwestern Italian Alps during two consecutive years. The results show that photosynthetic efficiency as assessed by fluorescence measurements was controlled by variations in air and soil temperature. Photosynthesis responded to variations in maximum air and soil temperature in a delayed way, with a varying lag depending on the seasonal period considered. The analysis of photosynthetic efficiency and pigment decline at the end of the growing season identified two senescence phases. During early senescence, plants manifested only the beginning of needle decolouration, while during late senescence pigment degradation led to a loss in photosynthetic efficiency. This behavior indicates that the beginning of needle yellowing and the decline in photosynthetic efficiency can occur at different times-a finding that should be considered in order to improve models of ecosystem processes.

Statistical optimization of simple culture conditions to produce biomass of an ochratoxigenic mould biocontrol yeast strain.

AIM: To maximize biomass production of an ochratoxigenic mould-controlling strain of Lachancea thermotolerans employing response surface methodology (RSM). METHODS AND RESULTS: Using Plackett-Burman screening designs (PBSD) and central composite designs (CCD), an optimized culture medium containing (g l(-1) ): fermentable sugars (FS), 139·2, provided by sugar cane molasses (CMz), (NH(4) )(2) HPO(4) (DAP), 9·0, and yeast extract (YE), 2·5, was formulated. Maximal cell concentration obtained after 24 h at 28°C was 24·2 g l(-1) cell dry weight (CDW). The mathematical model obtained was validated in experiments performed in shaken-flask cultures and also in aerated bioreactors. Maximum yield and productivity values achieved were, respectively, of 0·23 g CDW/g FS in a medium containing (g l(-1) ): FS, 87·0; DAP, 7·0; YE, 1·0; and of 0·96 g CDW l(-1) h(-1) in a medium containing (g l(-1) ): FS, 150·8 plus DAP, 6·9. CONCLUSIONS: Optimized culture conditions for maximizing yeast biomass production determined in flask cultures were applicable at a larger scale. The highest yield values were attained in media containing relatively low-CMz concentrations supplemented with DAP and YE. Yeast extract would not be necessary if higher productivity is the aim. SIGNIFICANCE AND IMPACT OF THE STUDY: Cells of L. thermotolerans produced aerobically could be sustainably produced in a medium just containing cheap carbon, nitrogen and phosphorus sources. Response surface methodology allowed the fine-tuning of cultural conditions.

The hyperspectral irradiometer, a new instrument for long-term and unattended field spectroscopy measurements.

Reliable time series of vegetation optical properties are needed to improve the modeling of the terrestrial carbon budget with remote sensing data. This paper describes the development of an automatic spectral system able to collect continuous long-term in-field spectral measurements of spectral down-welling and surface reflected irradiance. The paper addresses the development of the system, named hyperspectral irradiometer (HSI), describes its optical design, the acquisition, and processing operations. Measurements gathered on a vegetated surface by the HSI are shown, discussed and compared with experimental outcomes with independent instruments.

Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides.

AIMS: Analysis of the physiology and metabolism of Escherichia coli arcA and creC mutants expressing a bifunctional alcohol-acetaldehyde dehydrogenase from Leuconostoc mesenteroides growing on glycerol under oxygen-restricted conditions. The effect of an ldhA mutation and different growth medium modifications was also assessed. METHODS AND RESULTS: Expression of adhE in E. coli CT1061 [arcA creC(Con)] resulted in a 1.4-fold enhancement in ethanol synthesis. Significant amounts of lactate were produced during micro-oxic cultures and strain CT1061LE, in which fermentative lactate dehydrogenase was deleted, produced up to 6.5 +/- 0.3 g l(-1) ethanol in 48 h. Escherichia coli CT1061LE derivatives resistant to >25 g l(-1) ethanol were obtained by metabolic evolution. Pyruvate and acetaldehyde addition significantly increased both biomass and ethanol concentrations, probably by overcoming acetyl-coenzyme A (CoA) shortage. Yeast extract also promoted growth and ethanol synthesis, and this positive effect was mainly attributable to its vitamin content. Two-stage bioreactor cultures were conducted in a minimal medium containing 100 microg l(-1) calcium d-pantothenate to evaluate oxic acetyl-CoA synthesis followed by a switch into fermentative conditions. Ethanol reached 15.4 +/- 0.9 g l(-1) with a volumetric productivity of 0.34 +/- 0.02 g l(-1) h(-1). CONCLUSIONS: Escherichia coli responded to adhE over-expression by funnelling carbon and reducing equivalents into a highly reduced metabolite, ethanol. Acetyl-CoA played a key role in micro-oxic ethanol synthesis and growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Insight into the micro-oxic metabolism of E. coli growing on glycerol is essential for the development of efficient industrial processes for reduced biochemicals production from this substrate, with special relevance to biofuels synthesis.

European larch phenology in the Alps: can we grasp the role of ecological factors by combining field observations and inverse modelling?

Vegetation phenology is strongly influenced by climatic factors. Climate changes may cause phenological variations, especially in the Alps which are considered to be extremely vulnerable to global warming. The main goal of our study is to analyze European larch (Larix decidua Mill.) phenology in alpine environments and the role of the ecological factors involved, using an integrated approach based on accurate field observations and modelling techniques. We present 2 years of field-collected larch phenological data, obtained following a specifically designed observation protocol. We observed that both spring and autumn larch phenology is strongly influenced by altitude. We propose an approach for the optimization of a spring warming model (SW) and the growing season index model (GSI) consisting of a model inversion technique, based on simulated look-up tables (LUTs), that provides robust parameter estimates. The optimized models showed excellent agreement between modelled and observed data: the SW model predicts the beginning of the growing season (B(GS)) with a mean RMSE of 4 days, while GSI gives a prediction of the growing season length (L(GS)) with a RMSE of 5 days. Moreover, we showed that the original GSI parameters led to consistent errors, while the optimized ones significantly increased model accuracy. Finally, we used GSI to investigate interactions of ecological factors during springtime development and autumn senescence. We found that temperature is the most effective factor during spring recovery while photoperiod plays an important role during autumn senescence: photoperiod shows a contrasting effect with altitude decreasing its influence with increasing altitude.

Modelling the freezing response of baker's yeast prestressed cells: a statistical approach.

AIMS: To study the effect of prestress conditions on the freezing and thawing (FT) response of two baker's yeast strains and the use of statistical analysis to optimize resistance to freezing. METHODS AND RESULTS: Tolerance to FT of industrial strains of Saccharomyces cerevisiae was associated to their osmosensitivity and growth phase. Pretreatments with sublethal stresses [40 degrees C, 0.5 mol l(-1) NaCl, 1.0 mol l(-1) sorbitol or 5% (v/v) ethanol] increased freeze tolerance. Temperature or hyperosmotic prestresses increased trehalose contents, nevertheless no clear correlation was found with improved FT tolerance. Plackett-Burman design and response surface methodology were applied to improve freeze tolerance of the more osmotolerant strain. Optimal prestress conditions found were: 0.779 mol l(-1) NaCl, 0.693% (v/v) ethanol and 32.15 degrees C. CONCLUSIONS: Ethanol, saline, osmotic or heat prestresses increased freezing tolerance of two phenotypically distinct baker's yeast strains. A relationship among prestresses, survival and trehalose content was not clear. It was possible to statistically find optimal combined prestress conditions to increase FT tolerance of the osmotolerant strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Statistically designed combination of prestress conditions that can be applied during the production of baker's yeast could represent a useful tool to increase baker's yeast FT resistance.

Exploring the use of natural antimicrobial agents and pulsed electric fields to control spoilage bacteria during a beer production process.

Different natural antimicrobials affected viability of bacterial contaminants isolated at critical steps during a beer production process. In the presence of 1 mg/ml chitosan and 0.3 mg/ml hops, the viability of Escherichia coli in an all malt barley extract wort could be reduced to 0.7 and 0.1% respectively after 2 hour- incubation at 4 degrees C. The addition of 0.0002 mg/ml nisin, 0.1 mg/ml chitosan or 0.3 mg/ml hops, selectively inhibited growth of Pediococcus sp. in more than 10,000 times with respect to brewing yeast in a mixed culture. In the presence of 0.1 mg ml chitosan in beer, no viable cells of the thermoresistant strain Bacillus megaterium were detected. Nisin, chitosan and hops increased microbiological stability during storage of a local commercial beer inoculated with Lactobacillus plantarum or Pediococcus sp. isolated from wort. Pulsed Electric Field (PEF) (8 kV/cm, 3 pulses) application enhanced antibacterial activity of nisin and hops but not that of chitosan. The results herein obtained suggest that the use of these antimicrobial compounds in isolation or in combination with PEF would be effective to control bacterial contamination during beer production and storage.

Exploring the use of natural antimicrobial agents and pulsed electric fields to control spoilage bacteria during a beer production process / Exploración del uso de agentes antimicrobianos naturales y de campos eléctricos pulsantes para el control de bacterias contaminantes durante el proceso de elaboración de cerveza

Different natural antimicrobials affected viability of bacterial contaminants isolated at critical steps during a beer production process. In the presence of 1 mg/ml chitosan and 0.3 mg/ml hops, the viability of Escherichia coli in an all malt barley extract wort could be reduced to 0.7 and 0.1% respectively after 2 hour- incubation at 4 °C. The addition of 0.0002 mg/ml nisin, 0.1 mg/ml chitosan or 0.3 mg/ml hops, selectively inhibited growth of Pediococcus sp. in more than 10,000 times with respect to brewing yeast in a mixed culture. In the presence of 0.1mg ml chitosan in beer, no viable cells of the thermoresistant strain Bacillus megaterium were detected. Nisin, chitosan and hops increased microbiological stability during storage of a local commercial beer inoculated with Lactobacillus plantarum or Pediococcus sp. isolated from wort. Pulsed Electric Field (PEF) (8 kV/cm, 3 pulses) application enhanced antibacterial activity of nisin and hops but not that of chitosan. The results herein obtained suggest that the use of these antimicrobial compounds in isolation or in combination with PEF would be effective to control bacterial contamination during beer production and storage.

Diferentes antimicrobianos naturales disminuyeron la viabilidad de bacterias contaminantes aisladas en etapas críticas del proceso de producción de cerveza. En un extracto de malta, el agregado de 1 mg/ml de quitosano y de 0,3 mg ml de lúpulo permitió reducir la viabilidad de Escherichia coli a 0,7 y 0,1%, respectivamente, al cabo de 2 horas de incubación a 4 °C. El agregado de 0,0002 mg/ml de nisina, 0,1 mg/ml de quitosano o de 0,3 mg/ml de lúpulo inhibió selectivamente (10.000 veces más) el crecimiento de Pediococcus sp. respecto de la levadura de cerveza en un cultivo mixto. El agregado de 0,1 mg/ml de quitosano permitió disminuir la viabilidad de una cepa bacteriana termorresistente, Bacillus megaterium, hasta niveles no detectables. Por otra parte, el agregado de nisina, quitosano y lúpulo aumentó la estabilidad microbiológica durante el almacenamiento de cervezas inoculadas con Lactobacillus plantarum y Pediococcus sp. aislados de mosto de cerveza. La aplicación de campos eléctricos pulsantes (CEP) (3 pulsos de 8kV/cm) aumentó el efecto antimicrobiano de la nisina y del lúpulo, pero no el del quitosano. Los resultados obtenidos indicarían que el uso de antimicrobianos naturales en forma individual o en combinación con CEP puede constituir un procedimiento efectivo para el control de la contaminación bacteriana durante el proceso de elaboración y almacenamiento de la cerveza.

[Increase of rising activity of commercial yeasts by application of stress conditions during their propagation]. / Aumento de la actividad panificante de levaduras comerciales por aplicación de condiciones de estrés durante su propagación.

Rising activity determined as CO2 production of two commercial strains of Saccharomyces cerevisiae could be increased mainly in sweet bread doughs by introducing a "starvation/pulse feeding" schedule of sugar cane molasses during a fed-batch propagation. Such increase was strain dependent. Except for the trehalose intracellular level, other traits related to the yeast industrial performance were unaffected. Applicability of method for baker's yeast industrial production is discussed.

Aumento de la actividad panificante de levaduras comerciales por aplicación de condiciones de estrés durante su propagación / Increase of baking activity of commercial yeasts by application of stress conditions during their propagation

La actividad panificante valorada como producción de CO2 de dos cepas comerciales de Saccharomyces cerevisiae pudo ser incrementada, principalmente en amasijos azucarados, por la aplicación de un esquema de “hambreado/ pulso¼ de melaza de caña de azúcar durante su propagación bajo la forma de lote alimentado. Dicho incremento fue dependiente de la cepa utilizada. Otras características relacionadas con el comportamiento industrial de las levaduras no se vieron afectadas, con excepción de la concentración intracelular de trehalosa.Se discute la aplicabilidad del método para la producción industrial de levaduras de panificación.

Baking activity determined as CO2 production of two commercial strains of Saccharomyces cerevisiae could be increased mainly in sweet bread doughs by introducing a “starvation/ pulse feeding” schedule of sugar cane molasses during a fed-batch propagation . Such increase was strain dependent. Except for the trehalose intracellular level, other traits related to the yeast industrial performance were unaffected. Applicability of method for baker‘s yeast industrial production is discussed.

Aumento de la actividad panificante de levaduras comerciales por aplicación de condiciones de estrés durante su propagación. / [Increase of rising activity of commercial yeasts by application of stress conditions during their propagation]

Rising activity determined as CO2 production of two commercial strains of Saccharomyces cerevisiae could be increased mainly in sweet bread doughs by introducing a [quot ]starvation/pulse feeding[quot ] schedule of sugar cane molasses during a fed-batch propagation. Such increase was strain dependent. Except for the trehalose intracellular level, other traits related to the yeast industrial performance were unaffected. Applicability of method for bakers yeast industrial production is discussed.

Commercial baker's yeast stability as affected by intracellular content of trehalose, dehydration procedure and the physical properties of external matrices.

The effects of vacuum-drying and freeze-drying on the cell viability of a commercial baker's yeast, Saccharomyces cerevisiae, strain with different endogenous contents of trehalose were analyzed. An osmotolerant Zygosaccharomyces rouxii strain was used for comparative purposes. Higher viability values were observed in cells after vacuum-drying than after freeze-drying. Internal concentrations of trehalose in the range 10-20% protected cells in both dehydration processes. Endogenous trehalose concentrations did not affect the water sorption isotherm nor the Tg values. The effect of external matrices of trehalose and maltodextrin was also studied. The addition of external trehalose improved the survival of S. cerevisiae cells containing 5% internal trehalose during dehydration. Maltodextrin (1.8 kDa) failed to protect vacuum-dried samples at 40 degrees C. The major reduction in the viability during the freeze-drying process of the sensitive yeast cells studied was attributed to the freezing step. The suggested protective mechanisms for each particular system are vitrification and the specific interactions of trehalose with membranes and/or proteins. The failure of maltodextrins to protect cells was attributed to the fact that none of the suggested mechanisms of protection could operate in these systems.