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Characterization of antibiotic and biocide resistance genes and virulence factors of staphylococcus species associated with bovine mastitis in Rwanda

2020, Antók, Fruzsina Irén, Mayrhofer, Rosa, Marbach, Helene, Masengesho, Jean Claude, Keinprecht, Helga, Nyirimbuga, Vedaste, Fischer, Otto, Lepuschitz, Sarah, Ruppitsch, Werner, Ehling-Schulz, Monika, Feßler, Andrea T., Schwarz, Stefan, Monecke, Stefan, Ehricht, Ralf, Grunert, Tom, Spergser, Joachim, Loncaric, Igor

The present study was conducted from July to August 2018 on milk samples taken at dairy farms in the Northern Province and Kigali District of Rwanda in order to identify Staphylococcus spp. associated with bovine intramammary infection. A total of 161 staphylococcal isolates originating from quarter milk samples of 112 crossbred dairy cattle were included in the study. Antimicrobial susceptibility testing was performed and isolates were examined for the presence of various resistance genes. Staphylococcus aureus isolates were also analyzed for the presence of virulence factors, genotyped by spa typing and further phenotypically subtyped for capsule expression using Fourier Transform Infrared (FTIR) spectroscopy. Selected S. aureus were characterized using DNA microarray technology, multi-locus sequence typing (MLST) and whole-genome sequencing. All mecA-positive staphylococci were further genotyped using dru typing. In total, 14 different staphylococcal species were detected, with S. aureus being most prevalent (26.7%), followed by S. xylosus (22.4%) and S. haemolyticus (14.9%). A high number of isolates was resistant to penicillin and tetracycline. Various antimicrobial and biocide resistance genes were detected. Among S. aureus, the Panton–Valentine leukocidin (PVL) genes, as well as bovine leukocidin (LukM/LukF-P83) genes, were detected in two and three isolates, respectively, of which two also carried the toxic shock syndrome toxin gene tsst-1 bovine variant. t1236 was the predominant spa type. FTIR-based capsule serotyping revealed a high prevalence of non-encapsulated S. aureus isolates (89.5%). The majority of the selected S. aureus isolates belonged to clonal complex (CC) 97 which was determined using DNA microarray based assignment. Three new MLST sequence types were detected. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.

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Disturbing-free determination of yeast concentration in DI water and in glucose using impedance biochips

2020, Kiani, M., Du, N., Vogel, M., Raff, J., Hübner, U., Skorupa, I., Bürger, D., Schulz, S.E., Schmidt, O.G., Blaschke, D., Schmidt, H.

Deionized water and glucose without yeast and with yeast (Saccharomyces cerevisiae) of optical density OD600 that ranges from 4 to 16 has been put in the ring electrode region of six different types of impedance biochips and impedance has been measured in dependence on the added volume (20, 21, 22, 23, 24, 25 µL). The measured impedance of two out of the six types of biochips is strongly sensitive to the addition of both liquid without yeast and liquid with yeast and modelled impedance reveals a linear relationship between the impedance model parameters and yeast concentration. The presented biochips allow for continuous impedance measurements without interrupting the cultivation of the yeast. A multiparameter fit of the impedance model parameters allows for determining the concentration of yeast (cy) in the range from cy = 3.3 × 107 to cy = 17 × 107 cells/mL. This work shows that independent on the liquid, i.e., DI water or glucose, the impedance model parameters of the two most sensitive types of biochips with liquid without yeast and with liquid with yeast are clearly distinguishable for the two most sensitive types of biochips.

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Raman-spectroscopy based cell identification on a microhole array chip

2014, Neugebauer, U., Kurz, C., Bocklitz, T., Berger, T., Velten, T., Clement, J.H., Krafft, C., Popp, J.

Circulating tumor cells (CTCs) from blood of cancer patients are valuable prognostic markers and enable monitoring responses to therapy. The extremely low number of CTCs makes their isolation and characterization a major technological challenge. For label-free cell identification a novel combination of Raman spectroscopy with a microhole array platform is described that is expected to support high-throughput and multiplex analyses. Raman spectra were registered from regularly arranged cells on the chip with low background noise from the silicon nitride chip membrane. A classification model was trained to distinguish leukocytes from myeloblasts (OCI-AML3) and breast cancer cells (MCF-7 and BT-20). The model was validated by Raman spectra of a mixed cell population. The high spectral quality, low destructivity and high classification accuracy suggests that this approach is promising for Raman activated cell sorting.

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Staphylococcus aureus and methicillin resistant S. Aureus in nepalese primates: Resistance to antimicrobials, virulence, and genetic lineages

2020, Roberts, Marilyn C., Joshi, Prabhu Raj, Monecke, Stefan, Ehricht, Ralf, Müller, Elke, Gawlik, Darius, Diezel, Celia, Braun, Sascha D., Paudel, Saroj, Acharya, Mahesh, Khanal, Laxman, Koju, Narayan P., Chalise, Mukesh, Kyes, Randall C.

Staphylococcus aureus is a ubiquitous pathogen and colonizer in humans and animals. There are few studies on the molecular epidemiology of S. aureus in wild monkeys and apes. S. aureus carriage in rhesus macaques (Macaca mulatta) and Assam macaques (Macaca assamensis) is a species that has not previously been sampled and lives in remote environments with limited human contact. Forty Staphylococcus aureus isolates including 33 methicillin-susceptible S. aureus (MSSA) and seven methicillin-resistant S. aureus (MRSA) were characterized. Thirty-four isolates were from rhesus macaques and six isolates (five MSSA, one MRSA) were from Assam macaques. Isolates were characterized using StaphyType DNA microarrays. Five of the MRSA including one from Assam macaque were CC22 MRSA-IV (PVL+/tst+), which is a strain previously identified in Nepalese rhesus. One MRSA each were CC6 MRSA-IV and CC772 MRSA-V (PVL+). One MSSA each belonged to CC15, CC96, and CC2990. Six MRSA isolates carried the blaZ, while ten known CC isolates (seven MRSA, three MSSA) carried a variety of genes including aacA-aphD, aphA3, erm(C), mph(C), dfrA, msrA, and/or sat genes. The other 30 MSSA isolates belonged to 17 novel clonal complexes, carried no antibiotic resistance genes, lacked Panton–Valentine Leukocidin (PVL), and most examined exotoxin genes. Four clonal complexes carried egc enterotoxin genes, and four harbored edinB, which is an exfoliative toxin homologue. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

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Modified powder-in-tube technique based on the consolidation processing of powder materials for fabricating specialty optical fibers

2014, Auguste, J.-L., Humbert, G., Leparmentier, S., Kudinova, M., Martin, P.-O., Delaizir, G., Schuster, K., Litzkendorf, D.

The objective of this paper is to demonstrate the interest of a consolidation process associated with the powder-in-tube technique in order to fabricate a long length of specialty optical fibers. This so-called Modified Powder-in-Tube (MPIT) process is very flexible and paves the way to multimaterial optical fiber fabrications with different core and cladding glassy materials. Another feature of this technique lies in the sintering of the preform under reducing or oxidizing atmosphere. The fabrication of such optical fibers implies different constraints that we have to deal with, namely chemical species diffusion or mechanical stress due to the mismatches between thermal expansion coefficients and working temperatures of the fiber materials. This paper focuses on preliminary results obtained with a lanthano-aluminosilicate glass used as the core material for the fabrication of all-glass fibers or specialty Photonic Crystal Fibers (PCFs). To complete the panel of original microstructures now available by the MPIT technique, we also present several optical fibers in which metallic particles or microwires are included into a silica-based matrix.

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Microfluidic Network Simulations Enable On-Demand Prediction of Control Parameters for Operating Lab-on-a-Chip-Devices

2021, Böke, Julia Sophie, Kraus, Daniel, Henkel, Thomas

Reliable operation of lab-on-a-chip systems depends on user-friendly, precise, and predictable fluid management tailored to particular sub-tasks of the microfluidic process protocol and their required sample fluids. Pressure-driven flow control, where the sample fluids are delivered to the chip from pressurized feed vessels, simplifies the fluid management even for multiple fluids. The achieved flow rates depend on the pressure settings, fluid properties, and pressure-throughput characteristics of the complete microfluidic system composed of the chip and the interconnecting tubing. The prediction of the required pressure settings for achieving given flow rates simplifies the control tasks and enables opportunities for automation. In our work, we utilize a fast-running, Kirchhoff-based microfluidic network simulation that solves the complete microfluidic system for in-line prediction of the required pressure settings within less than 200 ms. The appropriateness of and benefits from this approach are demonstrated as exemplary for creating multi-component laminar co-flow and the creation of droplets with variable composition. Image-based methods were combined with chemometric approaches for the readout and correlation of the created multi-component flow patterns with the predictions obtained from the solver.

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Ultra-high sensitive strain sensor based on post-processed optical fiber Bragg grating

2014, Ferreira, M.S., Bierlich, J., Becker, M., Schuster, K., Santos, J.L., Frazão, O.

An ultra-high sensitive strain sensor is proposed. The sensing head, based on the post-processing of a fiber Bragg grating, is used to perform passive and active strain measurements. Both wavelength and full width half maximum dependences with the applied strain are studied for the passive sensor, where maximum sensitivities of 104.1 pm/με and 61.6 pm/με are respectively obtained. When combining the high performance of this sensor with a ring laser cavity configuration, the Bragg grating will act as a filter and high resolution measurements can be performed. With the proposed sensor, a resolution of 700 nε is achieved.

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Diffusion and interface effects during preparation of all-solid microstructured fibers

2014, Kobelke, J., Bierlich, J., Wondraczek, K., Aichele, C., Pan, Z., Unger, S., Schuster, K., Bartelt, H.

All-solid microstructured optical fibers (MOF) allow the realization of very flexible optical waveguide designs. They are prepared by stacking of doped silica rods or canes in complex arrangements. Typical dopants in silica matrices are germanium and phosphorus to increase the refractive index (RI), or boron and fluorine to decrease the RI. However, the direct interface contact of stacking elements often causes interrelated chemical reactions or evaporation during thermal processing. The obtained fiber structures after the final drawing step thus tend to deviate from the targeted structure risking degrading their favored optical functionality. Dopant profiles and design parameters(e.g., the RI homogeneity of the cladding) are controlled by the combination of diffusion and equilibrium conditions of evaporation reactions. We show simulation results of diffusion and thermal dissociation in germanium and fluorine doped silica rod arrangements according to the monitored geometrical disturbances in stretched canes or drawn fibers. The paper indicates geometrical limits of dopant structures in sub-μm-level depending on the dopant concentration and the thermal conditions during the drawing process. The presented results thus enable an optimized planning of the preform parameters avoiding unwanted alterations in dopant concentration profiles or in design parameters encountered during the drawing process.