ABSTRACT
Food spoilage is a routine challenge in food production. Saccharomyces cerevisiae is a major contaminating microorganism associated with fruit pulps and juices. Our study demonstrated the effect of a plasma corona discharge on S. cerevisiae viability, membrane permeability, and morphology when the cells were prepared in both dry and wet modes. The S. cerevisiae viability was examined as a function of the duration of plasma exposure, the sample's distance from the treating head, initial cell concentration, and yeast suspension volume. The results showed a linear correlation between the exposure duration and the CFU/mL in both dry and wet modes. When the initial yeast concentration was 106 CFU/mL, complete eradication in the dry and wet modes occurred after 45 and 240 s, respectively. Exposure of different initial concentrations of S. cerevisiae to plasma in dry (20 s) or wet (90 s) mode led to 2 to 3 orders of magnitude reduction. In both modes, there was total eradication when the initial cell concentration was about 103 CFU/mL. The cell-membrane permeability was examined using a flow cytometer and the fluorescent dye propidium iodide (PI). Plasma treatment in the dry mode for 30 and 45 s led to 51% and 76% PI-positive cells. Similar results were obtained in the wet mode but with a longer exposure for 120 and 240 s, respectively. Atmospheric plasma may provide disinfection technology for the food industry in a short process without heating.
ABSTRACT
Soil-borne pathogenic microorganisms are known to cause extensive crop losses. Agrobacterium tumefaciens, a member of the Proteobacteria, causes the neoplastic crown gall disease in plants. Plant protection is mainly based on toxic chemicals that are harmful to the environment. The use of cold atmospheric-pressure plasma is an attractive method for microbial eradication. Its antimicrobial mechanism includes the formation of large quantities of reactive oxygen species (ROS). The advantages of eradicating bacteria using cold plasma are not needed for chemicals, short treatment, and environmental temperatures. This study examined the impact of plasma corona discharge exposure on A. tumefaciens viability, membrane permeability, relative cell size, and ROS formation. The results showed that 90 s of plasma exposure led to a reduction by four orders of magnitude when the initial concentration was 1 × 107 CFU/mL and in a dry environment. When the initial concentration was 1 × 106 CFU/mL, 45 s of exposure resulted in total bacterial eradication. In a liquid environment, in an initial concentration of 2.02 × 106 CFU/mL, there was no complete bacterial eradication even at the most prolonged examined exposure (90 s). The influence of plasma treatment on the membrane permeability of A. tumefaciens, and their possible recovery, were analyzed using flow cytometer analysis using propidium iodide (PI). When the plasma-treated bacteria were suspended in Luria-Bertani (LB) (rich medium), the PI-positive count of the plasma-treated bacteria after two hours was 12 ± 3.9%. At the 24th hour, this percentage was only 1.74 ± 0.6%, as the control (0.7 ± 0.1%). These results may indicate the repair of the plasma-treated bacteria that were suspended in LB. At the 24th hour, the relative cell size of the treated bacteria shifted to the right, to ~3 × 104 forward side scatter (FSC), about 0.5-fold higher than the untreated cells. Measurement of the ROS showed that the intracellular fluorescence of the 90-s plasma-treated cells led to significant fluorescence formation of 32 relative fluorescence units (RFU)/cell (9 × 104 fold, compared to the nontreated cells). This study showed that cold plasma is a useful method for A. tumefaciens eradication. The eradication mechanism involves ROS generation, membrane permeability, and changes in cell size.
ABSTRACT
Crop contamination by soil-borne pathogenic microorganisms often leads to serious infection outbreaks. Plant protection requires disinfection of agricultural lands. The chemical and the physical disinfection procedures have several disadvantages, including an irreversible change in the soil ecosystem. Plasma, the "fourth state of matter" is defined as an ionized gas containing an equal number of negatively and positively charged particles. Cold-plasma technology with air or oxygen as the working gas generates reactive oxygen species, which are found to efficiently eradicate bacteria. In this study, we examined the effect of atmospheric plasma corona discharges on soil bacteria viability. Soil that was exposed to plasma for 60 s resulted in bacterial reduction by two orders of magnitude, from 1.1 × 105 to 2.3 × 103 cells g-1 soil. Exposure for a longer period of 5 min did not lead to further significant reduction in bacterial concentration (a final reduction of only 2.5 orders of magnitude). The bacterial viability was evaluated using a colorimetric assay based on the bacterial hydrogenases immediately after exposure and at selected times during 24 h. The result showed no recovery in the bacterial viability. Plasma discharged directly on bacteria that were isolated from the soil resulted in a reduction by four orders of magnitude in the bacterial concentration compared to untreated isolated bacteria: 2.6 × 10-3 and 1.7 × 10-7, respectively. The plasma-resistant bacteria were found to be related to the taxonomic phylum Firmicutes (98.5%) and comprised the taxonomic orders Bacillales (95%) and Clostridiales (2%). To our knowledge, this is the first study of soil bacteria eradication using plasma corona discharges.
ABSTRACT
Adiponectin is an adipose-derived hormone, with beneficial effects on insulin sensitivity and inflammation. The aim of this study was to clarify the autocrine/paracrine effects of globular adiponectin (gAd) administrated during differentiation on the function of the mature adipocytes. Experiments were performed on 3T3-L1 preadipocytes treated with gAd (10 nM), starting at an early stage of differentiation. gAd treatment during differentiation was without effect on mRNA expression of adiponectin and AdipoR2, but increased AdipoR1 expression. PPARgamma, perillipin and FABP4 mRNA expressions were lower in gAd-treated adipocytes, accompanied by a reduction in lipid accumulation. While mRNA expression of HSL was not affected by gAd compared to untreated adipocytes, both ATGL and FAS were reduced, indicating that gAd regulates both lipolysis and lipogenesis. PPARα, ACOX2 and UCPs mRNA expressions were not affected by gAd, indicating that the reduction in lipid content is not attributed to an increase in fatty-acid oxidation. In accord with the lower PPARγ expression, there was reduced Glut4 mRNA expression; however, insulin-induced PKB phosphorylation was enhanced by gAd, and glucose uptake was not altered. To investigate the effect of gAd on LPS-induced inflammatory response, cells were treated with gAd either during differentiation or 24 h before induction of inflammation in differentiated adipocytes. LPS-induced inflammatory response, as indicated by increase in IL6 and MCP-1 mRNA expression. gAd given through differentiation was much more effective in abrogating LPS-dependent cytokines production than gAd given to the mature adipocyte. In conclusion, elevated gAd at differentiation of 3T3-L1 cells leads to reduced lipid content, reduced lipid metabolism and high resistance to inflammation.
