Astrid H. Paulitsch-Fuchs

Alternating electric fields combined with activated carbon for disinfection of Gram negative and Gram positive bacteria in fluidized bed electrode system.

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.

Aqueous phenol and ethylene glycol solutions in electrohydrodynamic liquid bridging

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.

Effect of granular activated carbon concentration on the content of organic matter and salt, influencing E. coli activity and survival in fluidized bed disinfection reactor

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.

Behavioral study of selected microorganisms in an aqueous electrohydrodynamic liquid bridge

An aqueous electrohydrodynamic (EHD) floating liquid bridge is a unique environment for studying the influence of protonic currents (mA cm−2) in strong DC electric fields (kV cm−1) on the behavior of microorganisms. It forms in between two beakers filled with water when high-voltage is applied to these beakers. We recently discovered that exposure to this bridge has a stimulating effect on Escherichia coli.. In this work we show that the survival is due to a natural Faraday cage effect of the cell wall of these microorganisms using a simple 2D model. We further confirm this hypothesis by measuring and simulating the behavior of Bacillus subtilis subtilis, Neochloris oleoabundans, Saccharomyces cerevisiae and THP-1 monocytes. Their behavior matches the predictions of the model: cells without a natural Faraday cage like algae and monocytes are mostly killed and weakened, whereas yeast and Bacillus subtilis subtilis survive. The effect of the natural Faraday cage is twofold: First, it diverts the current from passing through the cell (and thereby killing it); secondly, because it is protonic it maintains the osmotic pressure in the cell wall, thereby mitigating cytolysis which would normally occur due to the low osmotic pressure of the surrounding medium. The method presented provides the basis for selective disinfection of solutions containing different microorganisms.

Different binarization processes validated against manual counts of fluorescent bacterial cells

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 (200× and 400×), staining procedures (DNA (Propidium Iodide) and RNA (FISH)) and substrates (polycarbonate filter or glass).

Microsatellite Typing of Catheter-Associated Candida albicans Strains

Candida albicans is the most common pathogenic fungus and occurs frequently in the digestive tract (Bernhardt, 1998; Doskey, 2004). Vaginal candidiasis (Mohanty et al. 2007; Paulitsch et al., 2006; Sobel, 2007) is also a wide spread problem. This species can become invasive, causing infections on many different sites in patients with severe underlying diseases (Marol & Yukesoy, 2008; Odds et al., 2007).

New staining methods for yeast like fungi under special consideration of human pathogenic fungi

A new method for in-cellular staining of yeast like fungi with Oregon Green and SYTOX Green is presented enabling their detection as well as the observation of cellular details via confocal laser scanning microscopy. Fluorochromes play an important role in many scientific disciplines including medicine, cell biology and botany. For the visualisation of fungal cell walls Calcofluor White is the flourochrome of choice. The necessity of an UV laser for its excitation makes it unpracticable for daily use. Safranin O, DAPI, 2NBDG, Ethidium Bromide and Acridin-orange are commonly used stains for nuclei in fugal microscopy. The attention was given to the possibility of using the differences in staining patterns to distinguish certain pathogenic yeast species e.g. Candida albicans and Candida krusei. Our results show that high quality microscopy of yeast like organisms can readily be achieved by the use of two suitable fluorochromes.