The present study had three goals: (i) to evaluate the relative quantities of aerosolized Bacillus atrophaeus spores deposited on the vertical, horizontal top, and horizontal bottom surfaces in a chamber; (ii) to assess the relative recoveries of the aerosolized spores from glass and stainless steel surfaces with a polyester swab and a macrofoam sponge wipe; and (iii) to estimate the relative recovery efficiencies of aerosolized B. atrophaeus spores and Pantoea agglomerans using a foam spatula at several different bacterial loads by aerosol distribution on glass surfaces. The majority of spores were collected from the bottom horizontal surface regardless of which swab type and extraction protocol were used. Swabbing with a macrofoam sponge wipe was more efficient in recovering spores from surfaces contaminated with high bioaerosol concentrations than swabbing with a polyester swab. B. atrophaeus spores and P. agglomerans culturable cells were detected on glass surfaces using foam spatulas when the theoretical surface bacterial loads were 2.88 × 10 4 CFU and 8.09 × 10 6 CFU per 100-cm 2 area, respectively. The median recovery efficiency from the surfaces using foam spatulas was equal to 9.9% for B. atrophaeus spores when the recovery was calculated relative to the theoretical surface spore load. Using a foam spatula permits reliable sampling of spores on the bioaerosol-exposed surfaces in a wide measuring range. The culturable P. agglomerans cells were recovered with a median efficiency of 0.001%, but staining the swab extracts with fluorescent dyes allowed us to observe that the viable cell numbers were higher by 1.83 log units than culturable organisms. However, additional work is needed to improve the analysis of the foam extracts in order to decrease the limit of detection of Bacillus spores and Gram-negative bacteria on contaminated surfaces.
1 April 2015 in International Biodeterioration & Biodegradation
This study was designed to evaluate the measuring range and lowest limit of detection of Bacillus endospores in the ambient room air when the Sartorius MD8 sampler, and two different culture methods for bacterial enumeration were used. Different concentrations of bioaerosol were generated inside the test chamber filled with either the high-efficiency particulate air (HEPA)-filtered air or with the ambient room air. The detection of endospores in the HEPA-filtered air was achievable: (1) when they were aerosolized at a concentration above 7.56 × 103 CFU/m3 and analyzed with spread plate method, and (2) when they were aerosolized at a concentration above 4.00 × 102 CFU/m3 and analyzed with pour plate method. The detection of endospores in the ambient room air was possible: (1) when they were aerosolized at a concentration above 9.1 × 103 CFU/m3 and analyzed with spread plate method, and (2) when they were aerosolized at a concentration above 5.6 × 102 CFU/m3 and analyzed with pour plate method. The microorganisms present in the ambient room air interfere with precise quantification of Bacillus endospores when their concentration is relatively low. The results of this study may be helpful in critical assessment of the results obtained from monitoring the air for bacterial endospores.
The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig TM3 cells. The cells were exposed to a moderate electric field (80 quasi-rectangular shape pulses, 60 ns pulse width, and an electric field of 14 kV/cm). The putative disturbances were recorded over 24 h. After exposure to the nanosecond pulsed electric field, a 19% increase in cell diameter, a loss of microvilli, and a 70% reduction in cell adhesion were observed. Some cells showed the nonapoptotic externalization of phosphatidylserine through the pores in the plasma membrane. The cell proportion in the subG1 phase increased by 8% at the expense of the S and G2/M phases, and the DNA was fragmented in a small proportion of the cells. The membrane mitochondrial potential and superoxide content decreased by 37% and 23%, respectively. Microarray’s transcriptome analysis demonstrated a negative transient effect on the expression of genes involved in oxidative phosphorylation, DNA repair, cell proliferation, and the overexpression of plasma membrane proteins. We conclude that nanosecond pulsed electric field affected the physiology and gene expression of TM3 cells transiently, with a noticeable heterogeneity of cellular responses.