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Isolation of bacteria from artificial bronchoalveolar lavage fluid using density gradient centrifugation and their accessibility by Raman spectroscopy
2021, Wichmann, Christina, Rösch, Petra, Popp, Jürgen
Raman spectroscopy is an analytical method to identify medical samples of bacteria. Because Raman spectroscopy detects the biochemical properties of a cell, there are many factors that can influence and modify the Raman spectra of bacteria. One possible influence is a proper method for isolation of the bacteria. Medical samples in particular never occur in purified form, so a Raman-compatible isolation method is needed which does not affect the bacteria and thus the resulting spectra. In this study, we present a Raman-compatible method for isolation of bacteria from bronchoalveolar lavage (BAL) fluid using density gradient centrifugation. In addition to measuring the bacteria from a patient sample, the yield and the spectral influence of the isolation on the bacteria were investigated. Bacteria isolated from BAL fluid show additional peaks in comparison to pure culture bacteria, which can be attributed to components in the BAL sample. The isolation gradient itself has no effect on the spectra, and with a yield of 63% and 78%, the method is suitable for isolation of low concentrations of bacteria from a complex matrix. Graphical abstract.
Noise Sources and Requirements for Confocal Raman Spectrometers in Biosensor Applications
2021, Jahn, Izabella J., Grjasnow, Alexej, John, Henry, Weber, Karina, Popp, Jürgen, Hauswald, Walter
Raman spectroscopy probes the biochemical composition of samples in a non-destructive, non-invasive and label-free fashion yielding specific information on a molecular level. Nevertheless, the Raman effect is very weak. The detection of all inelastically scattered photons with highest efficiency is therefore crucial as well as the identification of all noise sources present in the system. Here we provide a study for performance comparison and assessment of different spectrometers for confocal Raman spectroscopy in biosensor applications. A low-cost, home-built Raman spectrometer with a complementary metal-oxide-semiconductor (CMOS) camera, a middle price-class mini charge-coupled device (CCD) Raman spectrometer and a laboratory grade confocal Raman system with a deeply cooled CCD detector are compared. It is often overlooked that the sample itself is the most important “optical” component in a Raman spectrometer and its properties contribute most significantly to the signal-to-noise ratio. For this purpose, different representative samples: a crystalline silicon wafer, a polypropylene sample and E. coli bacteria were measured under similar conditions using the three confocal Raman spectrometers. We show that biosensor applications do not in every case profit from the most expensive equipment. Finally, a small Raman database of three different bacteria species is set up with the middle price-class mini CCD Raman spectrometer in order to demonstrate the potential of a compact setup for pathogen discrimination.
Particulate matter emissions during field application of poultry manure - The influence of moisture content and treatment
2021, Kabelitz, Tina, Biniasch, Oliver, Ammon, Christian, Nübel, Ulrich, Thiel, Nadine, Janke, David, Swaminathan, Senthilathiban, Funk, Roger, Münch, Steffen, Rösler, Uwe, Siller, Paul, Amon, Barbara, Aarnink, André J. A., Amon, Thomas
Along with industry and transportation, agriculture is one of the main sources of primary particulate matter (PM) emissions worldwide. Bioaerosol formation and PM release during livestock manure field application and the associated threats to environmental and human health are rarely investigated. In the temperate climate zone, field fertilization with manure seasonally contributes to local PM air pollution regularly twice per year (spring and autumn). Measurements in a wind tunnel, in the field and computational fluid dynamics (CFD) simulations were performed to analyze PM aerosolization during poultry manure application and the influence of manure moisture content and treatment. A positive correlation between manure dry matter content (DM) and PM release was observed. Therefore, treatments strongly increasing the DM of poultry manure should be avoided. However, high manure DM led to reduced microbial abundance and, therefore, to a lower risk of environmental pathogen dispersion. Considering the findings of PM and microbial measurements, the optimal poultry manure DM range for field fertilization was identified as 50–70%. Maximum PM10 concentrations of approx. 10 mg per m3 of air were measured during the spreading of dried manure (DM 80%), a concentration that is classified as strongly harmful. The modeling of PM aerosolization processes indicated a low health risk beyond a distance of 400 m from the manure application source. The detailed knowledge about PM aerosolization during manure field application was improved with this study, enabling manure management optimization for lower PM aerosolization and pathogenic release into the environment.