4,577
Views
4
CrossRef citations to date
0
Altmetric
Research Paper/Report

Removal of the cecum affects intestinal fermentation, enteric bacterial community structure, and acute colitis in mice

, , ORCID Icon &
Pages 218-235 | Received 12 Jun 2017, Accepted 17 Nov 2017, Published online: 13 Mar 2018

Figures & data

Figure 1. The experiment was designed as four factor factorial experiment with two levels of time (7 and 14 days post-inoculation [p.i.]), two levels of cecum (i.e. mice that underwent sham [Cec-] or cecectomy [Cec+] surgery), two levels of inflammation (mice that were orally administered phosphate buffered saline [CR-] or Citrobacter rodentium [CR+]), and two levels of butyrate (i.e. mice were administered phosphate buffered saline [But-] or butyrate [But+] via enemas every second day p.i.). Thus, 16 mice were included in each replicate, and four replicates were conducted on separate occasions equaling 64 mice in total.

Figure 1. The experiment was designed as four factor factorial experiment with two levels of time (7 and 14 days post-inoculation [p.i.]), two levels of cecum (i.e. mice that underwent sham [Cec-] or cecectomy [Cec+] surgery), two levels of inflammation (mice that were orally administered phosphate buffered saline [CR-] or Citrobacter rodentium [CR+]), and two levels of butyrate (i.e. mice were administered phosphate buffered saline [But-] or butyrate [But+] via enemas every second day p.i.). Thus, 16 mice were included in each replicate, and four replicates were conducted on separate occasions equaling 64 mice in total.

Figure 2. Concentration of: (A) total short chain fatty acids; (B) acetic acid; (C) butyric acid; (D) isobutyric acid; (E) valeric acid; (F) isovaleric acid; (G) propionic acid; and (H) caproic acid in feces from mice that received sham (Cec-) or cecectomy (Cec+) surgery and were orally administered PBS (CR-) or Citrobacter rodentium (CR+). Fatty acids were measured in feces collected prior to surgery (pre-surgery), between surgery and inoculation (post-surgery), and after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = 10 to 15 mice/treatment). *P ≤ 0.050, ***P ≤ 0.001.

Figure 2. Concentration of: (A) total short chain fatty acids; (B) acetic acid; (C) butyric acid; (D) isobutyric acid; (E) valeric acid; (F) isovaleric acid; (G) propionic acid; and (H) caproic acid in feces from mice that received sham (Cec-) or cecectomy (Cec+) surgery and were orally administered PBS (CR-) or Citrobacter rodentium (CR+). Fatty acids were measured in feces collected prior to surgery (pre-surgery), between surgery and inoculation (post-surgery), and after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = 10 to 15 mice/treatment). *P ≤ 0.050, ***P ≤ 0.001.

Figure 3. Alpha diversity of bacterial communities in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Vertical lines associated with histogram bars represent standard error of the means (n = six to eight mice/treatment).

Figure 3. Alpha diversity of bacterial communities in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Vertical lines associated with histogram bars represent standard error of the means (n = six to eight mice/treatment).

Figure 4. Principal component analysis based on Bray-Curtis dissimilarity in bacterial communities in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries 14 days post-inoculation.

Figure 4. Principal component analysis based on Bray-Curtis dissimilarity in bacterial communities in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries 14 days post-inoculation.

Figure 5. Relative abundance of bacteria genera within different families in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally adminstered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Bars indicate the mean (n = four mice/treatment).

Figure 5. Relative abundance of bacteria genera within different families in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally adminstered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Bars indicate the mean (n = four mice/treatment).

Figure 6. Relative abundance of genera within the Lachnospiraceae family in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Data represents the mean (n = four mice/treatment).

Figure 6. Relative abundance of genera within the Lachnospiraceae family in the proximal (A) and distal (B) colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Data represents the mean (n = four mice/treatment).

Figure 7. Relative abundance of genera within the Ruminococcaceae family in the proximal (A) and distal (B) colon of mice that received sham (Cec -) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Data represents the mean (n = four mice/treatment).

Figure 7. Relative abundance of genera within the Ruminococcaceae family in the proximal (A) and distal (B) colon of mice that received sham (Cec -) or cecectomy (Cec+) surgeries and were orally administered PBS (CR-) or Citrobacter rodentium (CR+) 14 days post-inoculation. Data represents the mean (n = four mice/treatment).

Figure 8. (A) Densities of viable C. rodentium (log CFU/g) in feces as determined by dilution plating for mice that received sham (Cec -) or cecectomy (Cec +) surgeries, were orally administered Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Vertical lines associated with markers indicate standard error of the means (n = four mice/treatment). (B-E) Visualization of C. rodentium in distal colon tissues at peak infection (i.e. 7 days post-inoculation); (B) Cec- and But-; (C) Cec- and But+; (D) Cec+ and But-; (E) Cec+ and But+. Cell nuclei are stained with DAPI (blue), total bacteria are stained with Alexa-488 (green) and γ-Proteobacteria (C. rodentium) are stained with Alexa-594 (red). Scale bar = 50 µm.

Figure 8. (A) Densities of viable C. rodentium (log CFU/g) in feces as determined by dilution plating for mice that received sham (Cec -) or cecectomy (Cec +) surgeries, were orally administered Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Vertical lines associated with markers indicate standard error of the means (n = four mice/treatment). (B-E) Visualization of C. rodentium in distal colon tissues at peak infection (i.e. 7 days post-inoculation); (B) Cec- and But-; (C) Cec- and But+; (D) Cec+ and But-; (E) Cec+ and But+. Cell nuclei are stained with DAPI (blue), total bacteria are stained with Alexa-488 (green) and γ-Proteobacteria (C. rodentium) are stained with Alexa-594 (red). Scale bar = 50 µm.

Figure 9. (A) Temporal change in body weight, and (B) change in body weight at 13 days after administration of Citrobacter rodentium or PBS. Mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or C. rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Vertical lines associated with markers or histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050.

Figure 9. (A) Temporal change in body weight, and (B) change in body weight at 13 days after administration of Citrobacter rodentium or PBS. Mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or C. rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Vertical lines associated with markers or histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050.

Figure 10. Histopathological scores of spleen tissues from mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010. The maximum possible score is 12.

Figure 10. Histopathological scores of spleen tissues from mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010. The maximum possible score is 12.

Figure 11. Histopathological scores of tissue sections from the (A) proximal and (B) distal colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010.

Figure 11. Histopathological scores of tissue sections from the (A) proximal and (B) distal colon of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS. Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010.

Figure 12. Expression of mRNA transcripts which code for: (A) Ifnγ; (B) Tnfα; (C) Il17a; (D) Il10; (E) Tgfβ; (F) Relmβ; (G) Muc2; (H) βd1; (I) Ffar2; and (J) Ffar3 in proximal colonic tissues of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS (p.i.). Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010.

Figure 12. Expression of mRNA transcripts which code for: (A) Ifnγ; (B) Tnfα; (C) Il17a; (D) Il10; (E) Tgfβ; (F) Relmβ; (G) Muc2; (H) βd1; (I) Ffar2; and (J) Ffar3 in proximal colonic tissues of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS (p.i.). Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010.

Figure 13. Expression of mRNA transcripts which code for: (A) Ifnγ; (B) Tnfα; (C) Il17a; (D) Il10; (E) Tgfβ; (F) Relmβ; (G) Muc2; (H) βd1; (I) Ffar2; and (J) Ffar3 in distal colonic tissues of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS (p.i.). Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010, ***P ≤ 0.001.

Figure 13. Expression of mRNA transcripts which code for: (A) Ifnγ; (B) Tnfα; (C) Il17a; (D) Il10; (E) Tgfβ; (F) Relmβ; (G) Muc2; (H) βd1; (I) Ffar2; and (J) Ffar3 in distal colonic tissues of mice that received sham (Cec-) or cecectomy (Cec+) surgeries, were orally administered PBS (CR-) or Citrobacter rodentium (CR+), and received enemas containing PBS (But-) or butyrate (But+). Data is shown 7 and 14 days after administration of C. rodentium or PBS (p.i.). Vertical lines associated with histogram bars represent standard error of the means (n = three to five mice/treatment). *P ≤ 0.050, **P ≤ 0.010, ***P ≤ 0.001.
Supplemental material

1408763_supp.pdf

Download PDF (396.7 KB)

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.