Inactivation of Campylobacter jejuni with dielectric barrier discharge plasma using air and nitrogen gases

Air and nitrogen gas are commonly used feed gases for plasma generation and are economically useful in industrial applications. The two gases were compared in dielectric barrier discharge plasma for the inactivation of Campylobacter jejuni on an agar surface. Plasma treatment with nitrogen gas for 20 s did not yield any reduction (p>0.05) in viable cell count. However, a 0.8-log reduction (p<0.05) in colony-forming units (CFU) occurred when the nitrogen gas was supplemented with 2% (vol/vol) air. The use of air only, air supplemented with 2% (vol/vol) nitrogen, or oxygen only further decreased the viable cell counts by 0.7-1.7-log CFU (p<0.05). These results suggest that oxygen in plasma generation is critically important for the increased inactivation effect. Scanning electron microscopy analysis showed much cell debris including fragmented flagella in the sample exposed to air plasma, while no cell debris was found in the sample exposed to nitrogen plasma. In transmission electron microscopy analysis, many C. jejuni cells exposed to air plasma had truncated flagella with sharp bends, while the cells exposed to nitrogen plasma were normal, strongly suggesting that the air plasma can reduce the virulence of C. jejuni. A BacLight assay showed that air plasma damaged the cellular membrane (p<0.05), whereas nitrogen plasma did not after 5- or 20-s treatment. The damage to the membrane was consistent with the reduced viable cell count. Based on confocal microscopic analysis, the similar results were found by visualizing the fluorescent-dye-stained cells. In addition, the prolonged nitrogen plasma for 2 min also damaged many cellular membranes. This study shows that air, especially oxygen, is more effective and destructive than nitrogen and provides evidence that membrane damage may be a major mechanism for the inactivation of C. jejuni exposed to plasma.