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. 2022 Jul 27;11(15):2234.
doi: 10.3390/foods11152234.

Effects of Cinnamon Essential Oil on Oxidative Damage and Outer Membrane Protein Genes of Salmonella enteritidis Cells

Zhen Zhang  1 , Yuanyuan Zhao  1 , Xueqin Chen  1 , Wei Li  1 , Wen Li  1 , Jianming Du  1 , Li Wang  1
Affiliations

Effects of Cinnamon Essential Oil on Oxidative Damage and Outer Membrane Protein Genes of Salmonella enteritidis Cells

Zhen Zhang et al. Foods. .

Abstract

Salmonella is an important pathogen causing food poisoning. Food safety and health are the themes of today′s society. As a class of food-borne pathogens, Salmonella enteritidis had become one of the common zoonotic pathogens. Cinnamon essential oil (CEO) had been reported as an antibacterial agent, but there are few studies on its antibacterial mechanism. This study investigated the effects of CEO on oxidative damage and outer membrane protein genes of Salmonella enteritidis cells. First, the reactive oxygen species content in bacteria treated with different concentrations of cinnamon essential oil was determined by fluorescence spectrophotometry, and the effects of superoxide dismutase (SOD), catalase (CAT) and superoxide dismutase (SOD), and catalase (CAT) and peroxidase (POD) were determined by the kit method. The activity of POD and the content of malondialdehyde (MDA) were investigated to investigate the oxidative damage of CEO to Salmonella enteritidis cells. By analyzing the effect of CEO on the Salmonella enteritidis cell membrane’s outer membrane protein gene expression, the mechanism of CEO′s action on the Salmonella enteritidis cell membrane was preliminarily discussed. The results showed that CEO treatment had an obvious oxidative damaging effect on Salmonella enteritidis. Compared with the control group, the increase in CEO concentration caused a significant increase in the bacteria ROS content. The observation technique experiment found that with the increase in CEO concentration, the number of stained cells increased, which indicated that CEO treatment would increase the ROS level in the cells, and it would also increase with the increase in CEO concentration, thus causing the oxidation of cells and damage. In addition, CEO treatment also caused the disruption of the balance of the cellular antioxidant enzymes (SOD, CAT, POD) system, resulting in an increase in the content of MDA, a membrane lipid metabolite, and increased protein carbonylation, which ultimately inhibited the growth of Salmonella enteritidis. The measurement results of cell membrane protein gene expression levels showed that the Omp genes to be detected in Salmonella enteritidis were all positive, which indicated that Salmonella enteritidis carried these four genes. Compared with the control group, the relative expressions of OmpF, OmpA and OmpX in the CEO treatment group were significantly increased (p < 0.05), which proved that the cell function was disturbed. Therefore, the toxicity of CEO to Salmonella enteritidis could be attributed to the damage of the cell membrane and the induction of oxidative stress at the same time. It was speculated that the antibacterial mechanism of CEO was the result of multiple effects. This work was expected to provide a theoretical basis for the development of new natural food preservatives and the prevention and control of Salmonella enteritidis.

Keywords: Salmonella enteritidis; cinnamon essential oil; outer membrane protein; oxidative damage.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of CEO treatment on intracellular ROS in S. enteritidis. (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, 2 MIC treatments).
Figure 2
Figure 2
Effects of CEO treatment on intracellular O2−·in S. enteritidis. (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, 2 MIC treatments).
Figure 3
Figure 3
Fluorescence microscope observation of the effect of CEO on ROS in S. enteritidis.
Figure 4
Figure 4
Effect on the enzyme activity of SOD in Salmonella enteritidis by CEO (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, and 2 MIC treatments).
Figure 5
Figure 5
Effect on the enzyme activity of CAT in Salmonella enteritidis by CEO (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, and 2 MIC treatments).
Figure 6
Figure 6
Effect on the enzyme activity of POD in Salmonella enteritidis by CEO. (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, and 2 MIC treatments).
Figure 7
Figure 7
Effect on the concentration of MDA in Salmonella enteritidis by CEO (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, and 2 MIC treatments).
Figure 8
Figure 8
Effects of CEO treatment on protein carbonylation content in S. enteritidis (note: a–c indicate significant differences (p < 0.05) among 1/2MIC, MIC, 2 MIC treatments).
Figure 9
Figure 9
Transcriptional analysis of the membrane protein genes in Salmonella enteritidis after CEO treatment (note: a–c indicate significant differences (p < 0.05) among 1/2 MIC, MIC, and 2 MIC treatments).
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