Plasmodium berghei ANKA causes intestinal malaria associated with dysbiosis - PubMed (original) (raw)

Eiji Miyauchi 3, Shota Nakamura 4, Makoto Hirai 5, Kazutomo Suzue 1, Takashi Imai 1, Takahiro Nomura 6, Tadashi Handa 7, Hiroko Okada 1, Chikako Shimokawa 1, Risa Onishi 1, Alex Olia 1, Jun Hirata 1, Haruyoshi Tomita 6, Hiroshi Ohno 3, Toshihiro Horii 8, Hajime Hisaeda 1

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Plasmodium berghei ANKA causes intestinal malaria associated with dysbiosis

Tomoyo Taniguchi et al. Sci Rep. 2015.

Erratum in

Abstract

Gastrointestinal symptoms, such as abdominal pain and diarrhea, are frequently observed in patients with Plasmodium falciparum malaria. However, the correlation between malaria intestinal pathology and intestinal microbiota has not been investigated. In the present study, infection of C57BL/6 mice with P. berghei ANKA (PbA) caused intestinal pathological changes, such as detachment of epithelia in the small intestines and increased intestinal permeability, which correlated with development with experimental cerebral malaria (ECM). Notably, an apparent dysbiosis occurred, characterized by a reduction of Firmicutes and an increase in Proteobacteria. Furthermore, some genera of microbiota correlated with parasite growth and/or ECM development. By contrast, BALB/c mice are resistant to ECM and exhibit milder intestinal pathology and dysbiosis. These results indicate that the severity of cerebral and intestinal pathology coincides with the degree of alteration in microbiota. This is the first report demonstrating that malaria affects intestinal microbiota and causes dysbiosis.

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Figures

Figure 1

Figure 1. Differences in pathology between B6 and BALB/c mice infected with _Pb_A.

Parasitemia (a) and survival (b) were monitored in B6 and BALB/c mice infected with _Pb_A. Parasitemia values are means ± S.D. from 20 B6 and 15 BALB/c mice. (c,d) Mice were injected with Evans blue dye 9 days after infection. Uninfected mice were used as controls. Brains of mice (c) and amounts of EB per g of brain (d) are shown. Data are means ± S.D. from 5 mice. Three repeated experiments showed similar results. Statistical analysis was performed using the Mann–Whitney _U_-test. **p < 0.05.

Figure 2

Figure 2. Appearance of intestines in mice infected with _Pb_A.

Representative photographs of whole intestines (a) and the length of small intestines (b) colons (c) and cecum (d) from _Pb_A-infected B6 and BALB/c mice at 9 days after infection are shown. Uninfected mice were used as controls. Each symbol represents an individual mouse, and bars indicate the mean of 5–10 mice. Three repeated experiments showed similar results. Statistical analysis was performed using Student’s unpaired _t_-test. **p < 0.01, ***p < 0.001.

Figure 3

Figure 3. Histological analyses of the small intestine in mice infected with _Pb_A.

Small or large intestines from B6 and BALB/c mice that were uninfected or 9 days after infection were stained with H&E (a,g) or alcian blue and nuclear fast red (b) respectively. Detachment of the intestinal epithelia (arrow) and sequestration of red blood cells in blood vessels (arrowheads) were observed. Length of villus (c) and depth of crypt (d) in the small intestine were measured. Each symbol indicates the mean of 100 villi or crypts from an individual mouse. Bars indicate the means of more than 5 mice. Thickness of mucus (e) and depth of crypts (f) in the colon were measured. Each symbol indicates the mean of 30 to 80 areas of mucus or 15 to 50 crypts from an individual mouse. (g) Representative micrographs of H&E-stained intestinal sections from _Pb_A-infected B6 mice indicate the destruction of villi and microscopic hemorrhage. (h) FITC-dextran concentrations in serum were measured in B6 mice that were uninfected or at 9 days after infection with _Pb_A. Each symbol indicates an individual mouse. Bars indicate the means of 5 or 8 control or _Pb_A-infected mice, respectively, from one experiment representative of the three experiments performed. Scale bars = 100 μm. Statistical analysis was performed using Student’s unpaired _t_-test. **p < 0.01, ***p < 0.001.

Figure 4

Figure 4. Temporal changes in intestinal microbiota in mice infected with _Pb_A.

(a) Principal coordinate analyses based on weighted UniFrac distance of fecal microbiota from B6 (n = 10) and BALB/c (n = 5) mice. Each symbol represents an individual mouse at different time periods and discriminated by different colors as indicated. (b) Mean weighted UniFrac distances between before and after _Pb_A infection in B6 and BALB/c mice are shown. Values are means ± S.D. from 10 B6 and 5 BALB/c mice. Statistical analysis was performed using two-way ANOVA with Bonferroni’s post hoc test. **p < 0.01 compared with uninfected mice.

Figure 5

Figure 5. Temporal changes in intestinal bacterial components in mice infected with _Pb_A.

(a) Relative abundance of fecal bacterial phyla in 5 B6 and 5 BALB/c mice. The numbers indicate individual mice, and each bar represents the results at the indicated time periods. (b–d) The relative abundance of the family of Lactobacillaceae (b), Enterobacteriaceae (c), and Verrucomicrobiaceae (d) in B6 and BALB/c mice infected with _Pb_A. Values are the means ± S.D. from 10 B6 and 5 BALB/c mice. Statistical analysis was performed using Dunnett’s test. **p < 0.01, ***p < 0.001 compared with uninfected mice.

Figure 6

Figure 6. Correlation between genera of bacteria and parasite burden in mice infected with _Pb_A.

(a) Representative co-plotted parasitemia and abundance of genera in 10 B6 and 5 BALB/c mice at 4 to 9 days after infection are shown. n and r denote the number of plots and the correlation coefficient, respectively. (b) The genera are listed in order of decreasing strength of correlation. Bright yellow or green indicates a stronger positive or negative correlation, respectively. Statistical analyses were performed using Pearson’s correlation coefficient test. *p < 0.05, **p < 0.01, ***p < 0.001.

References

    1. World Health Organization. World malaria report 2014. 2–6 (World Health Organization, 2014).
    1. Riley E. M. & Stewart V. A. Immune mechanisms in malaria: new insights in vaccine development. Nat Med 19, 168–178, 10.1038/nm.3083 (2013). -DOI -PubMed
    1. Reisinger E. C., Fritzsche C., Krause R. & Krejs G. J. Diarrhea caused by primarily non-gastrointestinal infections. Nat Clin Pract Gastroenterol Hepatol 2, 216–222, ncpgasthep0167 (2005). -PMC -PubMed
    1. Prasad R. N. & Virk K. J. Malaria as a cause of diarrhoea–a review. P N G Med J 36, 337–341 (1993). -PubMed
    1. Olsson R. A. & Johnston E. H. Histopathologic changes and small-bowel absorption in falciparum malaria. Am J Trop Med Hyg 18, 355–359 (1969). -PubMed

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