Effects of sepsis and its treatment measures on intestinal flora structure in critical care patients

Xiao-Juan Yang, Dan Liu, Hong-Yan Ren, Xiao-Ya Zhang, Jun Zhang, Xiao-Jun Yang, Xiao-Juan Yang, Dan Liu, Hong-Yan Ren, Xiao-Ya Zhang, Jun Zhang, Xiao-Jun Yang

Abstract

Background: Sepsis is a common disease in intensive care units, with high morbidity and mortality. Intestinal microecology plays a vital part in the development and progression of this disease, possibly because sepsis and its treatment cause specific changes in the composition of the intestinal flora.

Aim: To investigate the characteristics of intestinal flora disturbance in sepsis patients treated with antibiotics.

Methods: In this prospective comparative study, we enrolled ten patients with sepsis (sepsis group), hospitalized in the Department of Critical Care Medicine of the General Hospital, Ningxia Medical University, China (a class IIIa general hospital) from February 2017 to June 2017; ten patients without sepsis hospitalized in the same period (non-sepsis group) and ten healthy individuals (control group) were also enrolled. Fecal samples collected from the three groups were subjected to 16S rRNA gene sequencing and the intestinal flora diversity, structure, and composition were determined. Additionally, the dynamics of the intestinal flora diversity, structure, and composition in sepsis patients were investigated via 16S rRNA gene sequencing of samples collected 0 d, 3 d, and 7 d after admittance to the intensive care unit. Correlations between the serum levels of procalcitonin, endotoxin, diamine oxidase, and D-lactic acid and the intestinal flora composition of sepsis patients were also investigated.

Results: Compared with the healthy control group, sepsis and non-sepsis patients showed reduced intestinal flora α-diversity and a distinct flora structure, with Firmicutes as the dominant phylum, and significantly decreased proportions of Bacteroidetes, as well as Prevotella and Lachnospira, among other genera. Of note, the proportion of Enterococcus was significantly increased in the intestinal tract of sepsis patients. Interestingly, the α-diversity in the sepsis group decreased gradually, from days 1 to 7 of treatment. However, pairwise comparisons showed that both the diversity and structure of the intestinal flora were not significantly different considering the three different time points studied. Curiously, the serum levels of procalcitonin, endotoxin, diamine oxidase, and D-lactic acid in sepsis patients correlated with the prevalence of various bacterial genera. For example, the prevalence of Ruminococcus was positively correlated with serum procalcitonin, endotoxins, and diamine oxidase; similarly, the prevalence of Roseburia was positively correlated with serum procalcitonin, endotoxins, and D-lactic acid.

Conclusion: Sepsis patients in intensive care units show dysbiosis, lasting for at least 1 wk.

Keywords: 16S rRNA gene sequencing; Dynamic changes; Intestinal barrier index; Intestinal flora; Procalcitonin; Sepsis.

Conflict of interest statement

Conflict-of-interest statement: Yang XJ reports grants from Ministry of Science and Technology of the People's Republic of China, during the conduct of the study. Other authors have nothing to disclose.

©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.

Figures

Figure 1
Figure 1
α-diversity indexes of fecal microbiota in normal individuals, non-sepsis patients, and sepsis patients. A: ACE estimator; B: Chao 1 estimator; C: Shannon index; D: Simpson index.
Figure 2
Figure 2
Principal coordinate analysis of intestinal flora among normal individuals, sepsis patients, and non-sepsis patients. Principal coordinate analysis was plotted based on the Bray–Curtis dissimilarity between samples. The ellipses highlight the clustering of the fecal microbiomes according to groups (red: Healthy control group; blue: Non-sepsis group; green: Day 1 of sepsis).
Figure 3
Figure 3
Composition and comparison of the fecal microbiota among normal individuals, sepsis patients, and non-sepsis patients at the phylum level. A: Composition of the fecal microbiota at the phylum level among three groups; B: Comparison of the fecal microbiota at the phylum level among three groups. The average abundance of each phylum is depicted as the mean ± SE. P values were calculated using the Kruskal–Wallis rank-sum test: aP < 0.05, bP < 0.01.
Figure 4
Figure 4
Key phylotypes showing significant differences among normal individuals, sepsis patients, and non-sepsis patients. A: Comparison of the fecal microbiota at the genus level among the three groups. The average abundance of each genus is depicted as the mean ± SE. P values were calculated using the Kruskal–Wallis rank-sum test: aP < 0.05; bP < 0.01, cP < 0.001; B: Heatmap representing the relative abundance of differential operational taxonomic units (OTUs) among the three groups. The key OTUs whose relative abundance was above 1%, as per random forest-based models are displayed in the “figure”.
Figure 5
Figure 5
α-diversity indexes of the fecal microbiota in sepsis patients on days 1, 3, and 7 after admission to the intensive care unit vs healthy individuals. A: ACE estimator; B: Chao 1 estimator; C: Shannon index; D: Simpson index.
Figure 6
Figure 6
Principal coordinate analysis of the intestinal flora in sepsis patients on days 1, 3, and 7 after admission to the intensive care unit versus healthy individuals. Principal coordinate analysis was plotted based on the Bray–Curtis dissimilarity between samples. The ellipses highlight the clustering of the fecal microbiomes according to the groups (red: Healthy control group; blue: Sepsis patients on day 1; green: Sepsis patients on day 3; yellow: Sepsis patients on day 7).
Figure 7
Figure 7
Linear discriminant analysis scores indicating significant differences in the fecal microbiota between the sepsis patients within 1 wk after admission to the intensive care unit and the normal control group. Red: Enriched taxa in the healthy control group; blue: Enriched taxa in sepsis patients on day 1; green: Enriched taxa in sepsis patients on day 3; yellow: Enriched taxa in sepsis patients on day 7.
Figure 8
Figure 8
Correlation between clinical indicators and abundance of fecal microbiota in sepsis patients. Heatmap showing partial Spearman’s correlation coefficients among 30 genera and clinical indexes. Connecting lines represent the correlation coefficient values above 0.4 (red, positive correlation) or below -0.4 (blue, negative correlation). Solid lines represent P ≤ 0.01. Dotted lines represent 0.01 < P ≤ 0.05. The intensity of shading in the circles is proportional to the magnitude of the association. In the figure, “day” indicates the collection time of stool samples from sepsis patients after admission to the intensive care unit (ICU) (days 1, 3, and 7 after admission to the ICU). D-Lac: d-lactic acid; PCT: Procalcitonin; DAO: Diamine oxidase.

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