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Fig. 4
Effects of EcN on AIEC colonization
(A) Fish were added to 400 mL water containing 5 × 106 CFU/mL of only AIEC or 1:1 combination of AIEC and EcN for 6 h and then moved to fresh water for 18 h (24 hpi), 42 h (48 hpi), or 66 h (72 hpi). AIEC colonization in zebrafish intestine is shown on the y axis and is compiled from multiple experiments. Data are represented as mean +/− SEM from multiple experiments. Each dot represents the data from one fish. Total colonization per intestine was calculated after plating serial dilutions of intestinal homogenates 24 h postinfection. Statistical significance indicated above the data was determined by Student’s t test. ∗∗∗p < 0.0001, ∗∗p = 0.0019, and NS = nonsignificant.
(B) Fish were added to 400 mL water containing 5 × 106 CFU/mL of only AIEC or 1:1 combination of AIEC and EcN for 6 h and then moved to fresh water for up to 10 days with daily water changes. Three fish were sacrificed per day. The graph illustrates the significant decrease in AIEC colonization in AIEC and EcN (1:1) co-infected zebrafish compared with AIEC-infected zebrafish. The graph also illustrates successful EcN colonization in the zebrafish intestine over time. Error bars indicate standard deviation. Data shown are from three experiments.
(C) Representative H&E staining (5X) of zebrafish intestinal sections. Villus structure in AIEC-only and AIEC and EcN (1:1)-infected zebrafish were compared with the control zebrafish 24 hpi (i), 48 hpi (ii), and 72 hpi (iii). The H&E figures are representative of three independent experiments.