Assessment of oxygen kinetic parameters for closely related ammonia-oxidizing bacteria.

The reaction kinetics of lithotrophic ammonia-oxidizing bacteria (AOB) are strongly dependent on dissolved oxygen (DO) as their metabolism is an aerobic process. In this study, we estimate the kinetic parameters, including the oxygen affinity constant (Km[O2]) and the maximum oxygen consumption rate (Vmax[O2]), of different AOB species, by fitting the data to the Michaelis-Menten equation using nonlinear regression analysis. An example for three different species of Nitrosomonas bacteria (N.

Real-time stimulated Raman spectroscopy with a non-collinear optical parametric oscillator.

Ultrafast detection of microplastic particles is becoming a vital problem, as these particles are found in water sources worldwide. Ideally, a live analysis in flow is desirable to directly monitor the water quality for contaminations. Therefore, coherent Raman spectroscopy techniques require fast and broadband tunable lasers to address all relevant spectral regions of the investigated samples.

Calcium carbonate deposits and microbial assemblages on microplastics in oligotrophic freshwaters.

Microplastics are solid polymer particles with a wide variety of surface properties, found in most waterbodies, and known as carriers of distinct microbial communities affecting the fate of the particles in the environment. Little is known about the formation of mineral deposits on microplastics and how these deposits connect to microbial assemblages and affect the physicochemical properties of the particles. In addition, most of the available research on this topic is based on large microplastics with sizes between 100 μm and up to 5 mm, rather than the small microplastics often found in drinking water sources.

Small Sample Stress: Probing Oxygen-Deprived Ammonia-Oxidizing Bacteria with Raman Spectroscopy In Vivo.

The stress response of ammonia-oxidizing bacteria (AOB) to oxygen deprivation limits AOB growth and leads to different nitrification pathways that cause the release of greenhouse gases. Measuring the stress response of AOB has proven to be a challenge due to the low growth rates of stressed AOB, making the sample volumes required to monitor the internal stress response of AOB prohibitive to repeated analysis. In a proof-of-concept study, confocal Raman microscopy with excitation resonant to the heme c moiety of cytochrome c was used to compare the cytochrome c content and activity of stressed and unstressed (Nm 50), (Nm 57), (Nsp 10), and sp.

pH-Dependent Conformational Changes of KcsA Tetramer and Monomer Probed by Raman Spectroscopy.

KcsA is a tetrameric potassium channel formed out of four identical monomeric subunits used as a standard model for selective potassium transport and pH-dependent gating. Large conformational changes are reported for tetramer and monomer upon gating, and the response of the monomer being controversial with the two major studies partially contradicting each other. KcsA was analyzed as functional tetramers embedded in liposomes and as monomer subunits with confocal Raman microscopy under physiological conditions for the active and the closed channel state, using 532 nm excitation to avoid introducing conformational changes during the measurement.

Microplastics Detection in Streaming Tap Water with Raman Spectroscopy.

Microplastic particles have been found in drinking water sources worldwide and, thus, also in our food and beverages. Especially small microplastics, with sizes of 1 mm and less, cannot be identified reliably without spectroscopic means such as Fourier transform infrared spectroscopy (FTIR) or Raman spectroscopy, usually applied to the particles extracted from the samples. However, for drinking and tap water, with its comparatively low biological loads, direct observation may be possible and allows a point-of-entry monitoring for beverages and food to ensure uncontaminated drinking water is being used.

Simulation of Raman scattering including detector parameters and sampling volume.

Raman spectroscopy can be employed to measure the chemical composition of a sample, which can in turn be used to extract biological information. The aim of this paper is to introduce an efficient simulation technique for Raman spectroscopy in turbid (scattering) media taking into account relevant detector parameters and the sampling volume. We simulate the process of photon motion in turbid media by means of the Monte Carlo (MC) method.

Heat and Bleach: A Cost-Efficient Method for Extracting Microplastics from Return Activated Sludge.

The extraction of plastic microparticles, so-called microplastics, from sludge is a challenging task due to the complex, highly organic material often interspersed with other benign microparticles. The current procedures for microplastic extraction from sludge are time consuming and require expensive reagents for density separation as well as large volumes of oxidizing agents for organic removal, often resulting in tiny sample sizes and thus a disproportional risk of sample bias. In this work, we present an improved extraction method tested on return activated sludge (RAS).

Confocal Raman microscopy and fluorescent in situ hybridization - A complementary approach for biofilm analysis.

We combine confocal Raman microscopy (CRM) of wet samples with subsequent Fluorescent in situ hybridization (FISH) without significant limitations to either technique for analyzing the same sample of a microbial community on a cell-to-cell basis. This combination of techniques allows a much deeper, more complete understanding of complex environmental samples than provided by either technique alone. The minimalistic approach is based on laboratory glassware with micro-engravings for reproducible localization of the sample at cell scale combined with a fixation and de- and rehydration protocol for the respective techniques.

Temperature-sensitive gating of hCx26: high-resolution Raman spectroscopy sheds light on conformational changes.

The temperature-sensitive gating of human Connexin 26 (hCx26) was analyzed with confocal Raman microscopy. High-resolution Raman spectra covering the spectral range between 400 and 1500 rel. cm(-1) with a spectral resolution of 1 cm(-1) were fully annotated, revealing notable differences between the spectrum recorded from solubilized hCx26 in Ca(2+)-buffered POPC at 10°C and any other set of protein conditions (temperature, Ca(2+) presence, POPC presence).

Of microparticles and bacteria identification--(resonance) Raman micro-spectroscopy as a tool for biofilm analysis.

Confocal resonance Raman microscopy is a powerful tool for the non-invasive analysis of complex biological aggregates without preparation and prior knowledge of the samples. We present the capabilities of confocal resonance Raman microscopy with a spatial resolution of 350 nm2×2.0 μm and excitation times of 1 s and less per recorded spectrum.

Effects of ethanol, formaldehyde, and gentle heat fixation in confocal resonance Raman microscopy of purple nonsulfur bacteria.

Resonance Raman microscopy is well suited to examine living bacterial samples without further preparation. Therefore, comparatively little thought has been given to its compatibility with common fixation methods. However, fixation of cell samples is a very important tool in the microbiological sciences, allowing the preservation of samples in a specific condition for further examination, future measurements, transport, or later reference.