f Novel decontamination technologies, including cold low-pressure plasma and blue light (400 nm), are promising alternatives to conventional surface decontamination methods. However, the standardization of the assessment of such sterilization processes remains to be accomplished. Bacterial endospores of the genera Bacillus and Geobacillus are frequently used as biological indicators (BIs) of sterility. Ensuring standardized and reproducible BIs for reliable testing procedures is a significant problem in industrial settings. In this study, an electrically driven spray deposition device was developed, allowing fast, reproducible, and homogeneous preparation of Bacillus subtilis 168 spore monolayers on glass surfaces. A detailed description of the structural design as well as the operating principle of the spraying device is given. The reproducible formation of spore monolayers of up to 5 ؋ 10 7 spores per sample was verified by scanning electron microscopy. Surface inactivation studies revealed that monolayered spores were inactivated by UV-C (254 nm), low-pressure argon plasma (500 W, 10 Pa, 100 standard cubic cm per min), and blue light (400 nm) significantly faster than multilayered spores were. We have thus succeeded in the uniform preparation of reproducible, highly concentrated spore monolayers with the potential to generate BIs for a variety of nonpenetrating surface decontamination techniques. Microbial contamination on surfaces is a recurring problem within health, pharmaceutical, and food industry sectors (1, 2, 3). Thus, decontamination is a crucial step to ensure the sterility of food processing equipment, minimize spread of pathogens, and prevent the transmission of nosocomial infections (4). Common decontamination and disinfection procedures that are widely used for microbial inactivation include high temperatures, chemicals, or ionizing radiation (reviewed in reference 5). In order to ensure the efficiency and to validate the continuous functionality of a disinfection or sterilization procedure, biological testing standards are required. Bacterial spores are frequently used as a biological indicator (BI) of sterility, primarily because bacterial spores exhibit elevated resistance to chemical and physical methods of sterilization (6-11). Hence, a process that achieves full spore inactivation ensures complete elimination of other contaminating microorganisms.Variations in the performance of a BI have been reported repeatedly (12, 13). Besides variations in the intrinsic resistance properties of the microorganisms conferred, for instance, by variations in genetic traits or alteration of sporulation conditions (14), extrinsic factors also may affect the performance of BIs and, subsequently, the accurate determination of spore resistance and inactivation. For example, the sterilization results may be altered by poor choices of the carrier material for spore deposition (13,15) and, in particular, the BI manufacturing procedure (16). The method by which spores are mounted on carriers also is extremely impo...

A special focus area of planetary protection is the monitoring, control, and reduction of microbial contaminations that are detected on spacecraft components and hardware during and after assembly. In this study, wild-type spores of Bacillus pumilus SAFR-032 (a persistent spacecraft assembly facility isolate) and the laboratory model organism B. subtilis 168 were used to study the effects of low-pressure plasma, with hydrogen alone and in combination with oxygen and evaporated hydrogen peroxide as a process gas, on spore survival, which was determined by a colony formation assay. Spores of B. pumilus SAFR-032 and B. subtilis 168 were deposited with an aseptic technique onto the surface of stainless steel screws to simulate a spore-contaminated spacecraft hardware component, and were subsequently exposed to different plasmas and hydrogen peroxide conditions in a very high frequency capacitively coupled plasma reactor (VHF-CCP) to reduce the spore burden. Spores of the spacecraft isolate B. pumilus SAFR-032 were significantly more resistant to plasma treatment than spores of B. subtilis 168. The use of low-pressure plasma with an additional treatment of evaporated hydrogen peroxide also led to an enhanced spore inactivation that surpassed either single treatment when applied alone, which indicates the potential application of this method as a fast and suitable way to reduce spore-contaminated spacecraft hardware components for planetary protection purposes.

A transient spark micro-discharge in nitrogen is investigated between two sharpened electrodes at a pressure of 0.5 bar. The plasma parameters (gas temperature, electron density and reduced electric field) are determined using optical emission spectroscopy (OES) and numerical simulations. The gas temperature of 3500 ± 100 K is determined by the comparison of the measured and simulated rotational distributions of the photoemission spectra of neutral molecular nitrogen N 2 (C-B,0-0). Both direct and stepwise electron impact excitation are considered in the collision-radiative model. The rate constants for electron impact excitation processes are calculated for different electric field values using the electron velocity distribution function, which is simulated by solving the Boltzmann equation. The applied broadband echelle spectrometer is absolutely calibrated in a spectral range of 200 nm to 800 nm, using two standard light sources, a deuterium lamp and a tungsten ribbon lamp, which are certificated by the Physikalisch-Technische Bundesanstalt (PTB), Germany. With the aid of this absolutely calibrated echelle spectrometer and a microwave atmospheric plasma source operated in a nitrogen flow, the intensified charge-coupled device (ICCD) camera, provided with an in-house made optical arrangement for simultaneous two-wavelength diagnostic is calibrated. The spatial resolution of this diagnostic system under the studied plasma conditions amounts to 13 µm. The accurate examination of the experimental results allows determining the dominant process of electron impact excitation of molecular nitrogen ion from ionic ground state. Applying the chosen excitation model of the nitrogen photoemission, the spatially resolved reduced electric field and the electron density are determined. This is done by using the inverse Abel transformation of the absolute intensities of molecular nitrogen bands N 2 (C-B,0-0) and N + 2 (B-X,0-0), which were measured with the calibrated ICCD camera. The measured electric current of the micro-discharge is compared with the calculated one using the measured plasma parameters and a good coincidence is established.

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The results of a Multipole Resonance Probe (MRP) are compared to a Langmuir probe in measuring the electron density in Ar, H2, N2, and O2 mixtures. The MRP was designed for measurements in industry processes, i.e., coating or etching. To evaluate a possible influence on the MRP measurement due to molecular gases, different plasmas with increasing molecular gas content in a double inductively coupled plasma at 5 Pa and 10 Pa at 500 W are used. The determined electron densities from the MRP and the Langmuir probe slightly differ in H2 and N2 diluted argon plasmas, but diverge significantly with oxygen. In pure molecular gas plasmas, electron densities measured with the MRP are always higher than those measured with the Langmuir Probe, in particular, in oxygen containing mixtures. The differences can be attributed to etching of the tungsten wire in the Ar:O2 mixtures and rf distortion in the pure molecular discharges. The influence of a non-Maxwellian electron energy distribution function, negative ions or secondary electron emission seems to be of no or only minor importance.

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The development of new sterilization methods is still a major topic. The need for new techniques arises from the development of new instruments and the usage of different materials. Especially in the case of plastics with their beneficial properties, for example, in the field of implantology, plasma sterilization is seen as a promising alternative to the standard methods. However, 50 years after the first patent and although low‐pressure plasmas show excellent inactivation performance (>log 6 reduction), only one commercial system is available on the market for a distinct application. We will give a short review about known plasma sterilization mechanisms, the different plasma sterilization systems in use, analyze possible challenges for an industrial process and comment on possible solutions for a broader acceptance and utilization of low‐pressure plasma sterilization.