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. 2024 Apr 3;12(4):730.
doi: 10.3390/microorganisms12040730.

Inhibitory Effect of Lactococcus and Enterococcus faecalis on Citrobacter Colitis in Mice

Ullah Naveed  1   2   3 , Chenxi Jiang  1   2   3 , Qingsong Yan  1   2   3 , Yupeng Wu  1   2   3 , Jinhui Zhao  1   2   3 , Bowen Zhang  1   2   3 , Junhong Xing  1   2   3 , Tianming Niu  1   2   3 , Chunwei Shi  1   2   3 , Chunfeng Wang  1   2   3
Affiliations

Inhibitory Effect of Lactococcus and Enterococcus faecalis on Citrobacter Colitis in Mice

Ullah Naveed et al. Microorganisms. .

Abstract

Probiotics are beneficial for intestinal diseases. Research shows that probiotics can regulate intestinal microbiota and alleviate inflammation. Little research has been done on the effects of probiotics on colitis in mice. The purpose of this study was to investigate the inhibitory effect of the strains isolated and screened from the feces of healthy piglets on the enteritis of rocitrobacter. The compound ratio of isolated Lactobacillus L9 and Enterococcus faecalis L16 was determined, and the optimal compound ratio was selected according to acid production tests and bacteriostatic tests in vitro. The results showed that when the ratio of Lactobacillus L9 to Enterococcus faecalis L16 was 4:1, the pH value was the lowest, and the antibacterial diameter was the largest. Then, in animal experiments, flow cytometry was used to detect the number of T lymphocytes in the spleen and mesenteric lymph nodes of mice immunized with complex lactic acid bacteria. The results showed that the number of T lymphocytes in the spleen and mesenteric lymph nodes of mice immunized with complex lactic acid bacteria significantly increased, which could improve the cellular immunity of mice. The microbiota in mouse feces were sequenced and analyzed, and the results showed that compound lactic acid bacteria could increase the diversity of mouse microbiota. It stabilized the intestinal microbiota structure of mice and resisted the damage of pathogenic bacteria. The combination of lactic acid bacteria was determined to inhibit the intestinal colitis induced by Citrobacter, improve the cellular immune response of the body, and promote the growth of animals.

Keywords: Citrobacter rodentium; Lactobacillus combination; cellular immunity; intestinal microbiota.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Acid production and bacteriostatic activity of the Lactobacillus combination. 1:1, 1:2, 1:3, 1:4, 4:1, 3:1, and 2:1 in the figure indicate the ratios of L9 to L16. The compound acid production (A) and compound bacteriostatic activity (B). Data are represented as the mean ± SEM. Statistically significant differences are indicated (* p <0.05; ** p < 0.01; *** p < 0.001), and the line above the column marks the two groups with differences.
Figure 2
Figure 2
Changes in the body weight of mice after treatment (* p < 0.05).
Figure 3
Figure 3
Changes in the feed intake of mice after treatment.
Figure 4
Figure 4
Total antioxidant capacity in serum. Data are presented as the mean ± SEM of two experiments (n = 12 mice per group) (* p < 0.05).
Figure 5
Figure 5
Effects of the Lactobacillus combination on the body weight of mice after challenge (* p < 0.05).
Figure 6
Figure 6
Changes in the number of C. rodentium in faeces (*** p < 0.001).
Figure 7
Figure 7
Changes in the levels of the cytokines IFN-γ and IL-10. The content of IFN-γ in the serum of mice (A) and the content of IL-10 in the serum of mice (B). Data are presented as the mean ± SEM of two experiments (n = 12 mice per group) (* p < 0.05).
Figure 8
Figure 8
The histomorphological changes in the mouse colon and duodenum. Representative haematoxylin and eosin-stained sections of the colon (magnification: 400×). Pathological sections of the colon of mice in the PBS group (A), compound Lactobacillus group (B), and blank group (C).
Figure 9
Figure 9
T lymphocytes in the spleen and MLN were detected by flow cytometry. Results of flow cytometry in the spleen of mice in each group (A), statistical analysis of CD4+T lymphocytes and CD8+T lymphocytes in the spleen of mice in each group (B), MLN flow cytometry detection results of mice in each group (C), and statistical analysis of CD4+T lymphocytes and CD8+T lymphocytes in MLN of mice in each group (D). Data are presented as the mean ± SEM of two experiments (n = 3 mice per group) (* p < 0.05).
Figure 10
Figure 10
S-sequencing data of intestinal microflora of the mice in each group. Beta diversity analysis (A), species diversity analysis (B), analysis of the relative abundance of species (C), and LEfSe analysis at the genus level (D).
Figure 11
Figure 11
The function of intestinal flora in mice before and after infection with C. rodentium was compared.
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