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    Activity and electron donor preference of two denitrifying bacterial strains identified by Raman gas spectroscopy
    (Berlin [u.a.] : Springer, 2022) Blohm, Annika; Kumar, Swatantar; Knebl, Andreas; Herrmann, Martina; Küsel, Kirsten; Popp, Jürgen; Frosch, Torsten
    Human activities have greatly increased the input of reactive nitrogen species into the environment and disturbed the balance of the global N cycle. This imbalance may be offset by bacterial denitrification, an important process in maintaining the ecological balance of nitrogen. However, our understanding of the activity of mixotrophic denitrifying bacteria is not complete, as most research has focused on heterotrophic denitrification. The aim of this study was to investigate substrate preferences for two mixotrophic denitrifying bacterial strains, Acidovorax delafieldii and Hydrogenophaga taeniospiralis, under heterotrophic, autotrophic or mixotrophic conditions. This complex analysis was achieved by simultaneous identification and quantification of H2, O2, CO2, 14N2, 15N2 and 15N2O in course of the denitrification process with help of cavity-enhanced Raman spectroscopic (CERS) multi-gas analysis. To disentangle electron donor preferences for both bacterial strains, microcosm-based incubation experiments under varying substrate conditions were conducted. We found that Acidovorax delafieldii preferentially performed heterotrophic denitrification in the mixotrophic sub-experiments, while Hydrogenophaga taeniospiralis preferred autotrophic denitrification in the mixotrophic incubation. These observations were supported by stoichiometric calculations. The results demonstrate the prowess of advanced Raman multi-gas analysis to study substrate use and electron donor preferences in denitrification, based on the comprehensive quantification of complex microbial gas exchange processes. © 2021, The Author(s).
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    Rapid Raman spectroscopic analysis of stress induced degradation of the pharmaceutical drug tetracycline
    (Basel : MDPI, 2020) Domes, Christian; Frosch, Timea; Popp, Juergen; Frosch, Torsten
    Stress factors caused by inadequate storage can induce the unwanted degradation of active compounds in pharmaceutical formulations. Resonance Raman spectroscopy is presented as an analytical tool for rapid monitoring of small concentration changes of tetracycline and the metabolite 4-epianhydrotetracycline. These degradation processes were experimentally induced by changes in temperature, humidity, and irradiation with visible light over a time period of up to 23 days. The excitation wavelength ?exc = 413 nm was proven to provide short acquisition times for the simultaneous Raman spectroscopic detection of the degradation of tetracycline and production of its impurity in small sample volumes. Small concentration changes could be detected (down to 1.4% for tetracycline and 0.3% for 4-epianhydrotetracycline), which shows the potential of resonance Raman spectroscopy for analyzing the decomposition of pharmaceutical products. © 2020 by the authors.
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    Liquid-Core Microstructured Polymer Optical Fiber as Fiber-Enhanced Raman Spectroscopy Probe for Glucose Sensing
    (Washington, DC : OSA, 2020) Azkune, Mikel; Frosch, Timea; Arrospide, Eneko; Aldabaldetreku, Gotzon; Bikandi, Iñaki; Zubia, Joseba; Popp, Jürgen; Frosch, Torsten
    This work reports the development and application of two liquid-core microstructured polymer optical fibers (LC-mPOF) with different microstructure sizes. They are used in a fiber-enhanced Raman spectroscopy sensing platform, with the aim of detecting glucose in aqueous solutions in the clinically relevant range for sodium-glucose cotransporter 2 inhibitor therapy. The sensing platform is tested for low-concentration glucose solutions using each LC-mPOF. Results confirm that a significant enhancement of the Raman signal is achieved in comparison to conventional Raman spectroscopy. Additional measurements are carried out to obtain the valid measurement range, the resolution, and the limit of detection, showing that the LC-mPOF with 66-µm-diameter central hollow core has the highest potential for future clinical applications. Finally, preliminary tests successfully demonstrate glucose identification in urine. © 1983-2012 IEEE.
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    Grasshopper herbivory immediately affects element cycling but not export rates in an N‐limited grassland system
    (Ithaca, NY : ESA, 2021) Potthast, Karin; Meyer, Stefanie; Tischer, Alexander; Gleixner, Gerd; Sieburg, Anne; Frosch, Torsten; Michalzik, Beate
    As a cause of ecosystem disturbances, phytophagous insects are known to directly influence the element and organic matter (OM) cycling in ecosystems by their defoliation and excretion activity. This study focuses on the interplay between short-term, insect herbivory, plant responses to feeding activity, rhizosphere processes, and belowground nutrient availability under nutrient-poor soil conditions. To test the effects of insect herbivory on OM and nutrient cycling in an N-limited pasture system, mesocosm laboratory experiments were conducted using Dactylis glomerata as common grass species and Chorthippus dorsatus, a widespread grasshopper species, to induce strong defoliating herbivory. 13CO2 pulse labeling was used together with labeled 15N feces to trace the fate of C in soil respiration at the beginning of herbivory, and of C and N in above- and belowground plant biomass, grasshopper, feces, bulk soil, soil microbial biomass, throughfall solutions, and soil solutions. Within five days, herbivory caused a reduction in aboveground grass biomass by about 34%. A linear mixed-effects model revealed that herbivory significantly increased total dissolved C and N amounts in throughfall solutions by a factor of 4–10 (P < 0.05) compared with the control. In total, 27.6% of the initially applied feces 15N were translocated from the aboveground to the belowground system. A significant enrichment of 15N in roots led to the assumption that feces-derived 15N was rapidly taken up to compensate for the frass-related foliar N losses in light of N shortage. Soil microorganisms incorporated newly available 13C; however, the total amount of soil microbial biomass remained unaffected, while the exploitative grass species rapidly sequestered resources to facilitate its regrowth after herbivory attack. Heavy herbivory by insects infesting D. glomerata-dominated, N-deficient grasslands remarkably impacted belowground nutrient cycling by an instant amplification of available nutrients, which led to an intensified nutrient competition between plants and soil microorganisms. Consequently, these competitive plant–soil microbe interactions accelerated N cycling and effectively retained herbivory-mediated C and N surplus release resulting in diminished N losses from the system. The study highlighted the overarching role of plant adaptations to in situ soil fertility in short-term ecosystem disturbances.