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. 2021 Sep 24:12:737626.
doi: 10.3389/fmicb.2021.737626. eCollection 2021.

Inhibition of Quorum Sensing and Biofilm Formation of Esculetin on Aeromonas Hydrophila

Bing Sun  1   2 , Huaizhi Luo  1 , Huan Jiang  1 , Zhennan Wang  1 , Aiqun Jia  2
Affiliations

Inhibition of Quorum Sensing and Biofilm Formation of Esculetin on Aeromonas Hydrophila

Bing Sun et al. Front Microbiol. .

Abstract

Quorum sensing (QS) and biofilm formation inhibition activity of esculetin on Aeromonas hydrophila SHAe 115 were evaluated. Exposure to esculetin at 25, 50, and 100μg/ml significantly inhibited the production of protease and hemolysin, the formation of biofilms and attenuated the swarming motility of A. hydrophila SHAe 115. Biofilm forming inhibition was also observed through confocal laser scanning microscopy and scanning electron microscope. Quantitative real-time PCR analysis indicated that genes positively related to QS and biofilm formation were downregulated to varying degrees, while gene (litR) negatively related to biofilm formation was significantly upregulated. The phenotypic results were in good agreement with gene expression levels. These results indicated that esculetin would be a potential QS inhibitor for A. hydrophila.

Keywords: Aeromonas hydrophila SHAe 115; biofilm; esculetin; quantitative real-time PCR; quorum sensing.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Effect of esculetin at sub-MICs (25, 50, and 100μg/ml) on growth of Aeromonas hydrophila SHAe 115. The data represent the mean values of experiments performed in triplicates. Data are presented as the absorbance of mean±SD.
Figure 2
Figure 2
Effect of esculetin at sub-MICs (25, 50, and 100μg/ml) on hemolysin production of A. hydrophila SHAe 115. Data are presented as the absorbance of mean±SD of three independent experiments. ***p<0.001 compared to the DMSO control group by one-way ANOVA.
Figure 3
Figure 3
Effect of esculetin at sub-MICs (25, 50, and 100μg/ml) on protease activity of A. hydrophila SHAe 115. Data are presented as the absorbance of mean±SD of three independent experiments. ***p<0.001 compared to the DMSO control group by one-way ANOVA.
Figure 4
Figure 4
Effect of esculetin at sub-MICs (25, 50, and 100μg/ml) on swarming motility of A. hydrophila SHAe 115. (A) DMSO, (B) 25 μg/ml, (C) 50 μg/ml, (D) 100μg/ml, (E) Swarming zone diameter. Data are presented as the swarming diameter of mean±SD of three independent experiments. ***p<0.001 compared to the DMSO control group by one-way ANOVA.
Figure 5
Figure 5
Effect of esculetin on biofilm formation. (A) Quantitative analysis of biofilm biomass, (B) SEM images, and (C) CLSM images of A. hydrophila SHAe 115 biofilms treated with (a) DMSO, (b) 25μg/ml, (c) 50μg/ml, and (d) 100μg/ml of esculetin. Data are presented as the inhibition rate of mean±SD of three independent experiments. ***p<0.001 compared to the DMSO control group by one-way ANOVA.
Figure 6
Figure 6
Effect of esculetin at 100μg/ml on the expression of QS-related genes in A. hydrophila SHAe 115. Data are presented as the expression fold changes of mean±SD of three independent experiments. *p<0.05, **p<0.01, and ***p<0.001 compared to the DMSO control group by t-test.
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