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    Meteorological drivers of extremes in daily stem radius variations of beech, oak, and pine in Northeastern Germany: An event coincidence analysis
    (Lausanne : Frontiers Media, 2016) Siegmund, Jonatan F.; Sanders, Tanja G.M.; Heinrich, Ingo; van der Maaten, Ernst; Simard, Sonia; Helle, Gerhard; Donner, Reik V.
    Observed recent and expected future increases in frequency and intensity of climatic extremes in central Europe may pose critical challenges for domestic tree species. Continuous dendrometer recordings provide a valuable source of information on tree stem radius variations, offering the possibility to study a tree's response to environmental influences at a high temporal resolution. In this study, we analyze stem radius variations (SRV) of three domestic tree species (beech, oak, and pine) from 2012 to 2014. We use the novel statistical approach of event coincidence analysis (ECA) to investigate the simultaneous occurrence of extreme daily weather conditions and extreme SRVs, where extremes are defined with respect to the common values at a given phase of the annual growth period. Besides defining extreme events based on individual meteorological variables, we additionally introduce conditional and joint ECA as new multivariate extensions of the original methodology and apply them for testing 105 different combinations of variables regarding their impact on SRV extremes. Our results reveal a strong susceptibility of all three species to the extremes of several meteorological variables. Yet, the inter-species differences regarding their response to the meteorological extremes are comparatively low. The obtained results provide a thorough extension of previous correlation-based studies by emphasizing on the timings of climatic extremes only. We suggest that the employed methodological approach should be further promoted in forest research regarding the investigation of tree responses to changing environmental conditions.
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    Evaluating spatially resolved influence of soil and tree water status on quality of European plum grown in semi-humid climate
    (Lausanne : Frontiers Media, 2017) Käthner, Jana; Ben-Gal, Alon; Gebbers, Robin; Peeters, Aviva; Herppich, Werner B.; Zude-Sasse, Manuela
    In orchards, the variations of fruit quality and its determinants are crucial for resource effective measures. In the present study, a drip-irrigated plum production (Prunus domestica L. “Tophit plus”/Wavit) located in a semi-humid climate was studied. Analysis of the apparent electrical conductivity (ECa) of soil showed spatial patterns of sand lenses in the orchard. Water status of sample trees was measured instantaneously by means of leaf water potential, Ψleaf [MPa], and for all trees by thermal imaging of canopies and calculation of the crop water stress index (CWSI). Methods for determining CWSI were evaluated. A CWSI approach calculating canopy and reference temperatures from the histogram of pixels from each image itself was found to suit the experimental conditions. Soil ECa showed no correlation with specific leaf area ratio and cumulative water use efficiency (WUEc) derived from the crop load. The fruit quality, however, was influenced by physiological drought stress in trees with high crop load and, resulting (too) high WUEc, when fruit driven water demand was not met. As indicated by analysis of variance, neither ECa nor the instantaneous CWSI could be used as predictors of fruit quality, while the interaction of CWSI and WUEc did succeed in indicating significant differences. Consequently, both WUEc and CWSI should be integrated in irrigation scheduling for positive impact on fruit quality.
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    Strain engineering of ferroelectric domains in KxNa1−xNbO3 epitaxial layers
    (Lausanne : Frontiers Media, 2017) Schwarzkopf, Jutta; Braun, Dorothee; Hanke, Michael; Uecker, Reinhard; Schmidbauer, Martin
    The application of lattice strain through epitaxial growth of oxide films on lattice mismatched perovskite-like substrates strongly influences the structural properties of ferroelectric domains and their corresponding piezoelectric behavior. The formation of different ferroelectric phases can be understood by a strain-phase diagram, which is calculated within the framework of the Landau–Ginzburg–Devonshire theory. In this paper, we illustrate the opportunity of ferroelectric domain engineering in the KxNa1−xNbO3 lead-free material system. In particular, the following examples are discussed in detail: (i) Different substrates (NdGaO3, SrTiO3, DyScO3, TbScO3, and GdScO3) are used to systematically tune the incorporated epitaxial strain from compressive to tensile. This can be exploited to adjust the NaNbO3 thin film surface orientation and, concomitantly, the vector of electrical polarization, which rotates from mainly vertical to exclusive in-plane orientation. (ii) In ferroelectric NaNbO3, thin films grown on rare-earth scandate substrates, highly regular stripe domain patterns are observed. By using different film thicknesses, these can be tailored with regard to domain periodicity and vertical polarization component. (iii) A featured potassium concentration of x = 0.9 of KxNa1−xNbO3 thin films grown on (110) NdScO3 substrates favors the coexistence of two equivalent, monoclinic, but differently oriented ferroelectric phases. A complicated herringbone domain pattern is experimentally observed which consists of alternating MC and a1a2 domains. The coexistence of different types of ferroelectric domains leads to polarization discontinuities at the domain walls, potentially enabling high piezoelectric responses. In each of these examples, the experimental results are in excellent agreement with predictions based on the linear elasticity theory.
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    Sterilization of liquid foods by pulsed electric fields – an innovative ultra-high temperature process
    (Lausanne : Frontiers Media, 2015) Reineke, Kai; Schottroff, Felix; Meneses, Nicolas; Knorr, Dietrich
    The intention of this study was to investigate the inactivation of endospores by a combined thermal and pulsed electric field (PEF) treatment. Therefore, self-cultivated spores of Bacillus subtilis and commercial Geobacillus stearothermophilus spores with certified heat resistance were utilized. Spores of both strains were suspended in saline water (5.3 mS cm−1), skim milk (0.3% fat; 5.3 mS cm−1) and fresh prepared carrot juice (7.73 mS cm−1). The combination of moderate preheating (70–90°C) and an insulated PEF-chamber, combined with a holding tube (65 cm) and a heat exchanger for cooling, enabled a rapid heat up to 105–140°C (measured above the PEF chamber) within 92.2–368.9 μs. To compare the PEF process with a pure thermal inactivation, each spore suspension was heat treated in thin glass capillaries and D-values from 90 to 130°C and its corresponding z-values were calculated. For a comparison of the inactivation data, F-values for the temperature fields of both processes were calculated by using computational fluid dynamics (CFD). A preheating of saline water to 70°C with a flow rate of 5 l h−1, a frequency of 150 Hz and an energy input of 226.5 kJ kg−1, resulted in a measured outlet temperature of 117°C and a 4.67 log10 inactivation of B. subtilis. The thermal process with identical F-value caused only a 3.71 log10 inactivation. This synergism of moderate preheating and PEF was even more pronounced for G. stearothermophilus spores in saline water. A preheating to 95°C and an energy input of 144 kJ kg−1 resulted in an outlet temperature of 126°C and a 3.28 log10 inactivation, whereas nearly no inactivation (0.2 log10) was achieved during the thermal treatment. Hence, the PEF technology was evaluated as an alternative ultra-high temperature process. However, for an industrial scale application of this process for sterilization, optimization of the treatment chamber design is needed to reduce the occurring inhomogeneous temperature fields.
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    Bacterial community and PHB-accumulating bacteria associated with the wall and specialized niches of the hindgut of the forest cockchafer (Melolontha hippocastani)
    (Lausanne : Frontiers Media, 2017) Alonso-Pernas, Pol; Arias-Cordero, Erika; Novoselov, Alexey; Große, Christina; Rybak, Jürgen; Kaltenpoth, Martin; Westermann, Martin; Neugebauer, Ute; Boland, Wilhelm
    A characterization of the bacterial community of the hindgut wall of two larval and the adult stages of the forest cockchafer (Melolontha hippocastani) was carried out using amplicon sequencing of the 16S rRNA gene fragment. We found that, in second-instar larvae, Caulobacteraceae and Pseudomonadaceae showed the highest relative abundances, while in third-instar larvae, the dominant families were Porphyromonadaceae and Bacteroidales-related. In adults, an increase of the relative abundance of Bacteroidetes, Proteobacteria (γ- and δ- classes) and the family Enterococcaceae (Firmicutes) was observed. This suggests that the composition of the hindgut wall community may depend on the insect’s life stage. Additionally, specialized bacterial niches hitherto very poorly described in the literature were spotted at both sides of the distal part of the hindgut chamber. We named these structures “pockets.” Amplicon sequencing of the 16S rRNA gene fragment revealed that the pockets contained a different bacterial community than the surrounding hindgut wall, dominated by Alcaligenaceae and Micrococcaceae-related families. Poly-β-hydroxybutyrate (PHB) accumulation in the pocket was suggested in isolated Achromobacter sp. by Nile Blue staining, and confirmed by gas chromatography–mass spectrometry analysis (GC-MS) on cultured bacterial mass and whole pocket tissue. Raman micro-spectroscopy allowed to visualize the spatial distribution of PHB accumulating bacteria within the pocket tissue. The presence of this polymer might play a role in the colonization of these specialized niches.
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    High pressure thermal inactivation of Clostridium botulinum type E endospores – kinetic modeling and mechanistic insights
    (Lausanne : Frontiers Media, 2015) Lenz, Christian A.; Reineke, Kai; Knorr, Dietrich; Vogel, Rudi F.
    Cold-tolerant, neurotoxigenic, endospore forming Clostridium (C.) botulinum type E belongs to the non-proteolytic physiological C. botulinum group II, is primarily associated with aquatic environments, and presents a safety risk for seafood. High pressure thermal (HPT) processing exploiting the synergistic effect of pressure and temperature can be used to inactivate bacterial endospores. We investigated the inactivation of C. botulinum type E spores by (near) isothermal HPT treatments at 300–1200 MPa at 30–75°C for 1 s to 10 min. The occurrence of heat and lysozyme susceptible spore fractions after such treatments was determined. The experimental data were modeled to obtain kinetic parameters and represented graphically by isoeffect lines. In contrast to findings for spores of other species and within the range of treatment parameters applied, zones of spore stabilization (lower inactivation than heat treatments alone), large heat susceptible (HPT-induced germinated) or lysozyme-dependently germinable (damaged coat layer) spore fractions were not detected. Inactivation followed first order kinetics. Dipicolinic acid release kinetics allowed for insights into possible inactivation mechanisms suggesting a (poorly effective) physiologic-like (similar to nutrient-induced) germination at ≤450 MPa/≤45°C and non-physiological germination at >500 MPa/>60–70°C. Results of this study support the existence of some commonalities in the HPT inactivation mechanism of C. botulinum type E spores and Bacillus spores although both organisms have significantly different HPT resistance properties. The information presented here contributes to closing the gap in knowledge regarding the HPT inactivation of spore formers relevant to food safety and may help industrial implementation of HPT processing. The markedly lower HPT resistance of C. botulinum type E spores compared with the resistance of spores from other C. botulinum types could allow for the implementation of milder processes without endangering food safety.
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    Impact of different water activities (aw) adjusted by solutes on high pressure high temperature inactivation of Bacillus amyloliquefaciens spores
    (Lausanne : Frontiers Media, 2015) Sevenich, Robert; Reineke, Kai; Hecht, Philipp; Fröhling, Antje; Rauh, Cornelia; Schlüter, Oliver; Knorr, Dietrich
    Much research has been conducted to comprehend the mechanisms of high pressure (HP) inactivation of spores in aqueous systems but for food model systems these information are scarce. In these systems spores can interact with ingredients which then could possibly lead to retarded or reduced inactivation, which can cause a problem for the sterilization process. The protective mechanism of a reduced aw-value is still unclear. HP processing might prove valuable to overcome protective effects of solutes and achieve shorter process times for sterilization under HP. To gain insight into the underlying mechanisms five aw-values (0.9, 0.92, 0.94, 0.96, 1) were adjusted with two different solutes (NaCl, sucrose). Solutions were inoculated with spores of Bacillus amyloliquefaciens and treated at 105, 110, and 115°C at 600 MPa. Further a thermal inactivation was conducted at the same temperatures for a comparison with the HP data. Afterward, the influence of HP high temperature treatment on the inactivation, the dipicolinic acid (DPA)-release and membrane constitution was assessed by plate count, HPLC and flow cytometry (FCM). The results show that during HP treatments sucrose and salt both have a protective effect, in which the influence of sucrose on the retarded inactivation is higher. The threshold water activities (aw), which is 0.94, here salt and sucrose have a significant influence on the inactivation. The comparison of thermal (105–115°C) and HP and high temperature (600 MPa, 105–115°C) treated samples showed that the time needed to achieve a 4–5 log10 inactivation is reduced from 45 (aw = 1) to 75 (aw = 0.9) min at 105°C to 3 (aw = 1) to 15 (aw = 0.9) minutes at 600 MPa and 105°C. The release of DPA is the rate limiting step of the inactivation and therefore monitoring the release is of great interest. The DPA-release is slowed down in high concentrated solutions (e.g., sucrose, salt) in comparison to aw 1. Since there is a difference in the way the solutes protect the spore it could be seen as an inner spore membrane effect. Maybe as shown for vegetative microorganism the solutes can interact with membranes, e.g., the inner spore membrane. Flow cytometry (FCM) measurement data show a similar trend.
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    Impact of surface structure and feed gas composition on Bacillus subtilis endospore inactivation during direct plasma treatment
    (Lausanne : Frontiers Media, 2015) Hertwig, Christian; Steins, Veronika; Reineke, Kai; Rademacher, Antje; Klocke, Michael; Rauh, Cornelia; Schlüter, Oliver
    This study investigated the inactivation efficiency of cold atmospheric pressure plasma treatment on Bacillus subtilis endospores dependent on the used feed gas composition and on the surface, the endospores were attached on. Glass petri-dishes, glass beads, and peppercorns were inoculated with the same endospore density and treated with a radio frequency plasma jet. Generated reactive species were detected using optical emission spectroscopy. A quantitative polymerase chain reaction (qPCR) based ratio detection system was established to monitor the DNA damage during the plasma treatment. Argon + 0.135% vol. oxygen + 0.2% vol. nitrogen as feed gas emitted the highest amounts of UV-C photons and considerable amount of reactive oxygen and nitrogen species. Plasma generated with argon + 0.135% vol. oxygen was characterized by the highest emission of reactive oxygen species (ROS), whereas the UV-C emission was negligible. The use of pure argon showed a negligible emission of UV photons and atomic oxygen, however, the emission of vacuum (V)UV photons was assumed. Similar maximum inactivation results were achieved for the three feed gas compositions. The surface structure had a significant impact on the inactivation efficiency of the plasma treatment. The maximum inactivation achieved was between 2.4 and 2.8 log10 on glass petri-dishes and 3.9 to 4.6 log10 on glass beads. The treatment of peppercorns resulted in an inactivation lower than 1.0 log10. qPCR results showed a significant DNA damage for all gas compositions. Pure argon showed the highest results for the DNA damage ratio values, followed by argon + 0.135% vol. oxygen + 0.2% vol. nitrogen. In case of argon + 0.135% vol. oxygen the inactivation seems to be dominated by the action of ROS. These findings indicate the significant role of VUV and UV photons in the inactivation process of B. subtilis endospores.
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    β-adrenoceptor-mediated relaxation of urinary bladder muscle in β2-adrenoceptor knockout mice
    (Lausanne : Frontiers Media, 2016) Propping, Stefan; Lorenz, Kristina; Michel, Martin C.; Wirth, Manfred P.; Ravens, Ursula
    Background and Objective: In order to characterize the β-adrenoceptor (AR) subtypes involved in agonist-stimulated relaxation of murine urinary bladder we studied the effects of (-)-isoprenaline and CL 316,243 on tonic contraction and spontaneous contractions in detrusor strips of wild-type (WT) and β2-AR knockout (β2-AR KO) mice. Materials and Methods: Urinary bladders were isolated from male WT and β2-AR KO mice. β-AR subtype expression was determined with quantitative real-time PCR. Intact muscle strips pre-contracted with KCl (40 mM) were exposed to cumulatively increasing concentrations of (-)-isoprenaline or β3-AR agonist CL 316,243 in the presence and absence of the subtype-selective β-AR blockers CGP 20712A (β1-ARs), ICI 118,551 (β2-ARs), and L748,337 (β3-ARs). Results: Quantitative real-time PCR confirmed lack of β2-AR expression in bladder tissue from β2-AR KO mice. In isolated detrusor strips, pre-contraction with KCl increased basal tone and enhanced spontaneous activity significantly more in β2-AR KO than in WT. (-)-Isoprenaline relaxed tonic tension and attenuated spontaneous activity with similar potency, but the concentrations required were two orders of magnitude higher in β2-AR KO than WT. The concentration-response curves (CRCs) for relaxation were not affected by CGP 20712A (300 nM), but were shifted to the right by ICI 118,551 (50 nM) and L748,337 (10 μM). The -logEC50 values for (-)-isoprenaline in WT and β2-AR KO tissue were 7.98 and 6.00, respectively, suggesting a large receptor reserve of β2-AR. (-)-CL 316,243 relaxed detrusor and attenuated spontaneous contractions from WT and β2-AR KO mice with a potency corresponding to the drug’s affinity for β3-AR. L743,337 shifted the CRCs to the right. Conclusion: Our findings in β2-AR KO mice suggest that there is a large receptor reserve for β2-AR in WT mice so that this β-AR subtype will mediate relaxation of tone and attenuation of spontaneous activity under physiological conditions. Nevertheless, upon removal of this reserve, β3-AR can also mediate murine detrusor relaxation.
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    Amazon forest ecosystem responses to elevated atmospheric Co2 and alterations in nutrient availability: Filling the gaps with model-experiment integration
    (Lausanne : Frontiers Media, 2016) Hofhansl, Florian; Andersen, Kelly M.; Fleischer, Katrin; Fuchslueger, Lucia; Rammig, Anja; Schaap, Karst J.; Valverde-Barrantes, Oscar J.; Lapola, David M.
    The impacts of elevated atmospheric CO2 (eCO2) and alterations in nutrient availability on the carbon (C) storage capacity and resilience of the Amazon forest remain highly uncertain. Carbon dynamics are controlled by multiple eco-physiological processes responding to environmental change, but we lack solid experimental evidence, hampering theory development and thus representation in ecosystem models. Here, we present two ecosystem-scale manipulation experiments, to be carried out in the Amazon, that examine tropical ecosystem responses to eCO2 and alterations in nutrient availability and thus will elucidate the representation of crucial ecological processes by ecosystem models. We highlight current gaps in our understanding of tropical ecosystem responses to projected global changes in light of the eco-physiological assumptions considered by current ecosystem models. We conclude that a more detailed process-based representation of the spatial (e.g., soil type; plant functional type) and temporal (seasonal and inter-annual) variability of tropical forests is needed to enhance model predictions of ecosystem responses to projected global environmental change.