Fig 1.
Effect of microalgae on intestinal inflammation.
(A-G) Lateral view of 9 dpf Tg(BACmpo:GFP)i114 larvae after four days of feeding with the different diets; fishmeal (100FM), soybean meal (50SBM), and soybean meal + microalgae: 50SBM+Ch, 50SBM+Ts, 50SBM+A3Ng, 50SBM+Pt, or 50SBM+No. Black arrowheads indicate neutrophils. (H) The amount of intestinal neutrophils was quantified by immunohistochemistry against GFP. At least 25 larvae per diet were analyzed in each three different experiments. Statistical analysis was conducted using a non-parametric one-way ANOVA. ***P < 0.0001. Red bars represent the mean, and gray bars represent standard deviation.
Fig 2.
Effect of microalgae on fish response to Edwarsiella tarda infection.
Tg(BACmpo:GFP)i114 larvae fed with experimental or control diets were challenged with E. tarda for 5 h. Mortality was monitored every 12 h until 96 hour post challenge (hpc). Unchallenged larvae were subjected to the same feeding strategy and monitored during the same period. The graph represents the result obtained from three independent experiments. Statistical analysis was performed using survival curve analysis with Log-rank test against the 100FM and 50SBM diets. *P < 0.05, **P < 0.01, ***P < 0.001. Solid lines represent challenged larvae and dotted lines represent control (unchallenged) larvae.
Fig 3.
Microalgae improve neutrophil behavior after infection.
(A) Assay strategy. (B) Accumulated mortality in challenged (continuous line) and unchallenged (doted line) Tg(BACmpo:GFP)i114 larvae fed with 100FM or 50SBM with and without Tetraselmis sp. (Ts). Mortality was monitored every 12 h until 60 hours post challenge (hpc). (C) Amount of intestinal neutrophils before challenge (T0), and at 60 hpc (T60 Challenge). At least 25 larvae per condition were analyzed in each three independent experiments. Statistical analysis was conducted using a non-parametric Mann-Whitney test. *P < 0.5, ***P < 0.001. Red bars represent the mean, and gray bars represent standard deviation.
Table 1.
Ingredients and nutrient composition of control and experimental diets.
Table 2.
Control and experimental diets.