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). Microbial contamination on surfaces is a recurring problem within health, pharmaceutical, and food industry sectors ( 1, 2, 3). 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. 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. The reproducible formation of spore monolayers of up to 5 × 10 7 spores per sample was verified by scanning electron microscopy. A detailed description of the structural design as well as the operating principle of the spraying device is given. 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. Ensuring standardized and reproducible BIs for reliable testing procedures is a significant problem in industrial settings. Bacterial endospores of the genera Bacillus and Geobacillus are frequently used as biological indicators (BIs) of sterility. However, the standardization of the assessment of such sterilization processes remains to be accomplished. Novel decontamination technologies, including cold low-pressure plasma and blue light (400 nm), are promising alternatives to conventional surface decontamination methods.
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