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Periprosthetic infections are an eminent factor in patient care and also having significant economic implications. The number of biofilm-infection related replacement surgeries is increasing and will continue to do so in the following decades. To reduce both the health burden of the patients and the costs to the healthcare sector, new solutions for implant materials resistant to such infections are necessary. This study researches different surface modifications of cobalt-chromium-molybdenum (CoCrMo) based implant materials and their influence on the development of biofilms. Three smooth surfaces (CoCrMo, CoCrMo TiN, and CoCrMo polished) and three rough surfaces (CoCrMo porous coated, CoCrMo cpTi, and CoCrMo TCP) are compared. The most common infectious agents in periprosthetic infections are

Drinking water plumbing systems appear to be a unique environment for microorganisms as they contain few nutrients but a high mineral concentration. Interactions between mineral content and bacteria, such as microbial calcium carbonate precipitation (MCP) however, has not yet attracted too much attention in drinking water sector. This study aims to carefully examine MCP behavior of two drinking water bacteria species, which may potentially link scaling and biofouling processes in drinking water distribution systems. Evidence from cell density evolution, chemical parameters, and microscopy suggest that drinking water isolates can mediate CaCO3 precipitation through previously overlooked MCP mechanisms like ammonification or biosorption. The results also illustrate the active control of bacteria on the MCP process, as the calcium starts to concentrate onto cell surfaces only after reaching a certain cell density, even though the cell surfaces are shown to be the ideal location for the CaCO3 nucleation.

In this study, an alternative parameter for quantifying the signals of fluorescently labelled bacteria (e.g. propidium iodide, Cyanine 3, etc.) in microscopic images was investigated. Three common parameters (mean grey value (MGV), mean grey value which is corrected for the background (MGVcwB) and the signal to background ratio (SBR) per bacterial cell) are used as reference parameters. As an alternative, the coefficient of variation (CV) is defined as the ratio of the logarithm of the standard deviation and the logarithm of the mean grey value of a bacterial cell in a microscopic image. The actual fluorescence value was safeguarded by measuring commercially available fluorescence latex microspheres at regular time intervals within our study. The precision and the correlation of the respective values of MGV, MGVcwB, SBR and CV taken from identical images were measured and subsequently normalized in order to enhance the inter-parameter comparability. The average precision of CV was the highest (89% +/- 14) with decreasing numbers for MGVcwB, SBR, and MGV (78% +/- 25, 71% +/- 32, and, 52% +/- 22, respectively). Changes in operational parameters, e.g., microscope settings, protocol steps, etc., yielded good results for the CV but less precise results for MGV, MGVcwB, and SBR in the analyses of identical images.In conclusion, using the alternative parameter CV, changes in the composition of microbial ecosystems may thus be investigated at the highest precision level.

State of the art software methods (such as fixed value approaches or statistical approaches) to create a binary image of fluorescent bacterial cells are not as accurate and precise as they should be for counting bacteria and measuring their area. To overcome these bottlenecks, we introduce biological significance to obtain a binary image from a greyscale microscopic image. Using our biological significance approach we are able to automatically count about the same number of cells as an individual researcher would do by manual/visual counting. Using the fixed value or statistical approach to obtain a binary image leads to about 20% less cells in automatic counting. In our procedure we included the area measurements of the bacterial cells to determine the right parameters for background subtraction and threshold values. In an iterative process the threshold and background subtraction values were incremented until the number of particles smaller than a typical bacterial cell is less than the number of bacterial cells with a certain area. This research also shows that every image has a specific threshold with respect to the optical system, magnification and staining procedure as well as the exposure time. The biological significance approach shows that automatic counting can be performed with the same accuracy, precision and reproducibility as manual counting. The same approach can be used to count bacterial cells using different optical systems (Leica, Olympus and Navitar), magnification factors (200x and 400x), staining procedures (DNA (Propidium Iodide) and RNA (FISH)) and substrates (polycarbonate filter or glass). (C) 2016 Elsevier B.V. All rights reserved.

The authors wish to make the following corrections to this paper [1]: In all instances mentioning “2 kG·m−1”, these gradients should be replaced by “770 G·m−1 (WCM 62081992) and 740 G·m−1 (WCM 62083545)”: Abstract, page 1, line 4; Chapter 1 [...]

Granular activated carbon (GAC) is used in water treatment systems, typically to remove pollutants such as natural organic matter, volatile organic compounds, chlorine, taste, and odor. GAC is also used as a key component of a new technology that combines a fluidized bed reactor with radio frequency electric fields for disinfection. So far, the effects of GAC on bacteria in these fluidized bed reactors are unclear. This paper describes a systematic study of the physico-chemical changes in five microbial media compositions caused by different concentrations (23–350?g/L) of GAC, and the effects of these physico-chemical changes on the metabolic activity and survival of a model microorganism (Escherichia coli YMc10) in a fluidized bed reactor. The chemical adsorption taking place in suspensions with specific GAC changed nutritional, osmotic, and pH conditions in the investigated microbial media (LB, diluted LB, PBS, diluted PBS, and tap water), leading to a decay of the metabolic activity and survival of E. coli. Especially media that are poor in organic and mineral compounds (e.g., PBS) with suspended GAC showed a concentration decay of 3.5?Log CFU/mL E. coli after 6?h. Organic compounds depletion and severe pH variation were enhanced in the presence of higher GAC concentrations. Biotechnol. Bioeng. 2014;111: 2009–2018.

Strong electric fields for disinfection of wastewaters have been employed already for several decades. An innovative approach combining low strength (7 V/cm) alternating electric fields with a granular activated carbon fluidized bed electrode (FBE) for disinfection was presented recently. For disinfection performance of FBE several pure microbial cultures were tested: Bacillus subtilis, Bacillus subtilis subsp. subtilis, Enterococcus faecalis as representatives from Gram positive bacteria and Erwinia carotovora, Pseudomonas luteola, Pseudomonas fluorescens and Escherichia coli YMc10 as representatives from Gram negative bacteria. The alternating electric field amplitude and shape were kept constant. Only the effect of alternating electric field frequency on disinfection performance was investigated. From the bacteria tested, the Gram negative strains were more susceptible and the Gram positive microorganisms were more resistant to FBE disinfection. The collected data indicate that the efficiency of disinfection is frequency and strain dependent. During 6 h of disinfection, the decrease above 2 Log units was achieved with P. luteola and E. coli at 10 kHz and at dual frequency shift keying (FSK) modulated signal with frequencies of 10 kHz and 140 kHz. FBE technology appears to offer a new way for selective bacterial disinfection, however further optimizations are needed on treatment duration, and energy input, to improve effectiveness.

When a high-voltage direct-current is applied to two beakers filled with water or polar liquid dielectrica, a horizontal bridge forms between the two beakers. This experiment was first carried out by Lord Armstrong in 1893 and then forgotten until recently. Such bridges are stable by the action of electrohydrodynamic (EHD) forces caused by electric field gradients counteracting gravity. Due to these gradients a permanent pumping of liquid from one beaker into the other is observed. At macroscopic scale several of the properties of a horizontal water bridge can be explained by modern electrohydrodynamics, analyzing the motion of fluids in electric fields. Whereas on the molecular scale water can be described by quantum mechanics, there is a conceptual gap at mesoscopic scale which is bridged by a number of theories including quantum mechanical entanglement and coherent structures in water – theories that we discuss here. Much of the phenomenon is already understood, but even more can still be learned from it, since such “floating” liquid bridges resemble a small high voltage laboratory of their own: The physics of liquids in electric fields of some kV/cm can be studied, even long time experiments like neutron or light scattering are feasible since the bridge is in a steady-state equilibrium and can be kept stable for hours. It is also an electro-chemical reactor where compounds are transported through by the EHD flow, enabling the study of electrochemical reactions under potentials which are otherwise not easily accessible. Last but not least the bridge provides the experimental biologist with the opportunity to expose living organisms such as bacteria to electric fields without killing them, but with a significant influence on their behavior, and possibly, even on their genome.

In 2012 Coey proposed a theory on the mechanism of magnetic water treatment based on the gradient of the applied field rather than its absolute strength. We tested this theory by measuring the effect of very weak field magnets (≤ 10 G) containing strong magnetic inhomogeneities (ΔB = 770 G·m−1 (WCM 62081992) and 740 G·m−1 (WCM 62083545)) on tap water samples by the use of electric impedance spectroscopy (EIS) and laser scattering. Our results show an increased formation of nm-sized prenucleation clusters (dynamically ordered liquid like oxyanion polymers or “DOLLOPs”) due to the exposure to the magnetic field and thus are consistent with Coey’s theory which is therefore also applicable to very weak magnetic fields as long as they contain strong gradients.

The formation of aqueous bridges containing phenol and ethylene glycol as well as bisphenol-A, hydrochinone and p-cresol under the application of high voltage DC (“liquid bridges”) is reported. Detailed studies were made for phenol and glycol with concentrations from 0.005 to 0.531 mol L−1. Conductivity as well as substance and mass transfers through these aqueous bridges are discussed and compared with pure water bridges. Previously suggested bidirectional mass transport is confirmed for the substances tested. Anodic oxidation happens more efficiently when phenol or glycol are transported from the cathode to the anode since in this case the formation of a passivation layer or electrode poisoning are retarded by the electrohydrodynamic (EHD) flow. The conductivity in the cathode beaker decreases in all experiments due to electrophoretic transport of naturally dissolved carbonate and bicarbonate to the anode. The observed electrochemical behavior is shortly discussed and compared to known mechanisms.

Granular activated carbon (GAC) is used in water treatment systems, typically to remove pollutants such as natural organic matter, volatile organic compounds, chlorine, taste, and odor. GAC is also used as a key component of a new technology that combines a fluidized bed reactor with radio frequency electric fields for disinfection. So far, the effects of GAC on bacteria in these fluidized bed reactors are unclear. This paper describes a systematic study of the physico-chemical changes in five microbial media compositions caused by different concentrations (23-350 g/L) of GAC, and the effects of these physico-chemical changes on the metabolic activity and survival of a model microorganism (Escherichia coli YMc10) in a fluidized bed reactor. The chemical adsorption taking place in suspensions with specific GAC changed nutritional, osmotic, and pH conditions in the investigated microbial media (LB, diluted LB, PBS, diluted PBS, and tap water), leading to a decay of the metabolic activity and survival of E. coli. Especially media that are poor in organic and mineral compounds (e.g., PBS) with suspended GAC showed a concentration decay of 3.5 Log CFU/mL E. coli after 6 h. Organic compounds depletion and severe pH variation were enhanced in the presence of higher GAC concentrations.

Biofilms may interfere with membrane performance in at least three ways: (i) increase of the transmembrane pressure drop, (ii) increase of feed channel (feed-concentrate) pressure drop, and (iii) increase of transmembrane passage. Given the relevance of biofouling, it is surprising how few data exist about the hydraulic resistance of biofilms that may affect the transmembrane pressure drop and membrane passage. In this study, biofilms were generated in a lab scale cross flow microfiltration system at two fluxes (20 and 100 L m−2 h−1) and constant cross flow (0.1 m s−1). As a nutrient source, acetate was added (1.0 mg L−1 acetate C) besides a control without nutrient supply. A microfiltration (MF) membrane was chosen because the MF membrane resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric substances (EPS) were determined, it was demonstrated that no internal membrane fouling occurred and that the fouling layer actually consisted of a grown biofilm and was not a filter cake of accumulated bacterial cells. At 20 L m−2 h−1 flux with a nutrient dosage of 1 mg L−1 acetate C, the resistance after 4 days reached a value of 6×1012 m−1. At 100 L m−2 h−1 flux under the same conditions, the resistance was 5×1013 m−1. No correlation of biofilm resistance to biofilm thickness was found; Biofilms with similar thickness could have different resistance depending on the applied flux. The cell number in biofilms was between 4×107 and 5×108 cells cm−2. At this number, bacterial cells make up less than a half percent of the overall biofilm volume and therefore did not hamper the water flow through the biofilm significantly. A flux of 100 L m−2 h−1 with nutrient supply caused higher cell numbers, more biomass, and higher biofilm resistance than a flux of 20 L m−2 h−1. However, the biofilm thickness after 4 days at a flux of 100 L m−2 h−1 (97 μm) was in the same order of magnitude as the thickness of a biofilm at a flux of 20 L m−2 h−1 (114 μm). An increase of flux caused an increased biofilm resistance while a decrease of flux caused a decreased resistance. The effect was reversible. It is suggested that the biofilm resistance is mainly attributed to EPS, probably due to the tortuosity (“hair-in-sink-effect”) of the biopolymers to water molecules travelling across the biofilm. The data show clearly that biofilm resistance (6×1012 m−1) was high compared to the intrinsic resistance of the employed MF membrane (5×1011 m−1). However, in nanofiltration (intrinsic membrane resistance ca. 2×1013 m−1) and reverse osmosis membranes (intrinsic resistance ca. 9×1013 m−1), the biofilm will not contribute significantly to the overall resistance.

When a high-voltage direct-current is applied to two beakers filled with water, a horizontal electrohydrodynamic (EHD) bridge forms between the two beakers. In this work we study the transport and behavior of bacterial cells added to an EHD bridge set-up. Organisms were added to one or to both beakers, and the transport of the cells through the bridge was monitored using optical and microbiological techniques. It is shown that Escherichia coli top10 (Invitrogen, Carlsbad, CA, USA) and bioluminescent E. coli YMC10 with a plasmid (pJE202) containing Vibrio fischeri genes can survive the exposure to an EHD liquid bridge set-up and the cells are drawn toward the anode due to their negative surface charge. Dielectrophoresis and hydrostatic forces are likely to be the cause for their transport in the opposite direction which was observed as well, but to a much lesser extent. Most E. coli YMC10 bacteria which passed the EHD bridge exhibited increased luminescent activity after 24 h. This can be explained by two likely mechanisms: nutrient limitation in the heavier inoculated vials and a 'survival of the strongest' mechanism.

We report on the infrared emission of aqueous bridges under the application of high dc voltage (‘floating water bridge’) over the range between 400 and 2500 cm−1 (4.0–10.3 µm). Comparison with bulk water of the same temperature reveals an additional broad peak at ∼2200 cm−1 as well as water vapour emission lines. Two complementary explanations are presented for the broad peak: first, a cooperative proton transfer comprising an orientational motion along the direction of conduction is suggested. Second, the electrolysis-less current flow is explained by a proton/defect-proton band mechanism, which is in line with the cooperative proton transfer. The water vapour emissions occur due to collision ionization of space charges with micro- and nano-droplets which are electrosprayed from the liquid/gas interface.

Here we demonstrate the proof-of-principle that a coaxial stub resonator can be used to detect early stages of biofilm formation. After promising field tests using a stub resonator with a stainless steel inner conductor as sensitive element, the sensitivity of the system was improved by using a resonator of shorter physical length, implying higher resonance frequencies (and by that a higher frequency range of operation) and improved sensitivity towards dispersion. In addition, the space between inner and outer conductor was filled up with glass beads, thereby exploiting the larger surface area available for biofilm formation.Analysis of the biofilm and the stub resonator signal, both as function of time, indicates that the sensor allows detection of early stages of biofilm formation. In addition, the sensor signal clearly discriminates between the first stages of biofilm formation (characterized by separated, individual spots of bacterial growth on the glass beads) and the presence of a nearly homogeneous biofilm later on in time. Model simulations based on the transmission line theory predict a shift of the sensor response in the same direction and order of magnitude as observed in the biofouling experiments, thereby confirming the operating principle of the sensor.

Syntrophobacter fumaroxidans strain MPOBT is the best-studied species of the genus Syntrophobacter. The species is of interest because of its anaerobic syntrophic lifestyle, its in-volvement in the conversion of propionate to acetate, H2 and CO2 during the overall degra-dation of organic matter, and its release of products that serve as substrates for other microor-ganisms. The strain is able to ferment fumarate in pure culture to CO2 and succinate, and is also able to grow as a sulfate reducer with propionate as an electron donor. This is the first complete genome sequence of a member of the genus Syntrophobacter and a member genus in the family Syntrophobacteraceae. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 4,990,251 bp long genome with its 4,098 protein-coding and 81 RNA genes is a part of the Microbial Genome Program (MGP) and the Genomes to Life (GTL) Program project.