Figure 1.
Physiologic and 16S Changes Related to LPS Instillation Directly into Lungs of Mice.
A) Body weight changed following LPS instillation, P<0.01 from 24 hours through 96 hours versus baseline. B) Change in total cells in the bronchoalveolar (BAL) fluid following LPS instillation, P<0.05 from 48 and 72 hours versus baseline. C) Cecal bacterial load following LPS instillation, P<0.05 at 24 hours versus baseline. D) Blood bacterial load following LPS instillation, P<0.05 at 4, 48, and 72 hours versus baseline. E) Total bacterial load in the BAL fluid following LPS instillation. F) Sourcetracker analysis of the baseline BAL similarity to the blood bacterial microbiota from 4 to 168 hours post LPS instillation.
Figure 2.
Changes in Lung, Blood, and Cecum Following LPS Instillation in Mice Treated and Not Treated with Antibiotics.
A) Total cells in the BAL following LPS instillation, P<0.05 at 24 hours for the neomycin group versus LPS alone group. B) Total PMN in the BAL fluid following LPS instillation, P<0.05 at 24 hours for the neomycin group versus LPS alone group. C) Percentage of PMNs in the BAL fluid following LPS instillation. D) Total bacterial load in the cecum following LPS instillation. No significant difference was found between groups (P>0.05) E) Total bacterial load in the BAL following LPS instillation, P<0.0001 for the LPS alone group compared with neomycin or streptomycin group. F) Total bacterial load in the blood following LPS instillation, P<0.05 at 4 hours for the LPS alone group compared with neomycin or streptomycin group.
Figure 3.
Overall Shannon Diversity by Group.
Overall Shannon diversity measures for all samples in the study, P<0.05 for the first three groups versus the extraction negative and negative controls.
Figure 4.
Shannon Diversity by Treatment Group and Location.
A) Shannon diversity by time and treatment group in the cecum. B) Shannon diversity by time and treatment group within the BAL, P<0.05 at 4 hours for the LPS group. C) Shannon diversity by time and treatment group within the blood, P<0.05 at baseline for the neomycin group.
Figure 5.
Constrained Principle Coordinate Analysis (PCoA) of the Cecum microbiota.
A) PCoA analysis of the time course experiment, PERMANOVA, P<0.05. Red are the baseline control mice (n = 5), green are all the PBS timepoints combined (4–168 hours), and blue represents the LPS groups. The labels in the LPS group represent the central point for each timepoint within the LPS group B) PCoA analysis of the antibiotic experiment, PERMANOVA, P<0.05. Neomycin baseline is in grey, neomycin at 4 hours is in blue, and neomycin at 24 hours is in red. Streptomycin baseline is in yellow, streptomycin at 4 hours is in purple, and streptomycin at 24 hours is in green. The labels represent the central point for each cluster.
Figure 6.
Prominent OTU correlations to Blood Bacterial Load.
A) Linear regression of bacterial load in the blood and Phyllobacteriaceae (OTU 40) relative abundance, Y = 0.0037X – 1.2 (R2 = 0.65). B) Result on Phyllobacteriaceae relative abundance after antibiotic treatment and amount in the control samples, P<0.05 for Phyllobacteriaceae against all groups. C) Overall total reads detected in the blood for Phyllobacteriaceae with either PBS or LPS instillation. Overall P-value for the row factor (LPS versus PBS) was 0.015 and the overall P-value for the column factor (time points) was 0.0025. Post hoc analysis showed no significant difference between the groups. D) Phyllobacteriaceae after 4 weeks of either Streptomycin or Neomycin pre-treatment.