FIGURE SUMMARY
Title

Pichia pastoris composition expressed aerolysin mutant of Aeromonas veronii as an oral vaccine evaluated in zebrafish (Danio rerio)

Authors
Yao, Y.Y., Zhang, Q.S., Liu, S.B., Yang, H.W., Chen, X.Y., Yang, Y.L., Gao, C.C., Ran, C., Teame, T., Zhang, Z., Zhou, Z.G.
Source
Full text @ Mar Life Sci Technol
Fig. 1

Validation of the recombinant P. pastoris GS115-NTaer strain and safety of the antigen. A PCR analysis of P. pastoris genome and WB analysis of proteins secreted into medium. B Hemolysis test on blood plate. C Hemolysis rate of different concentrations of aerolysin Hmaer and its mutant NTaer. In two-group comparisons, significant differences were marked with ** (P < 0.01) or **** (P < 0.0001)
Fig. 2

Growth performance of zebrafish fed with control diet, GS115 or GS115-NTaer vaccine. A Initial body weight. B Final body weight. C Weight gain (%). D Feed conversion rate (FCR). E Survival rate (%). Data represent the means (± SD) of six replicates of each group, significant differences would be marked with different letter when P < 0.05
Fig. 3

Effect of vaccine on gut epidermal physical barrier and immunological response. A–D Relative expression of genes associated with tight junctions. E–G Content of lysozyme (LYZ), complement C3 and C4 in gut. H–N Profile of immune-related gene expression post vaccination. Data represent the means (± SD) of six replicates of each group, groups with different letters indicate significant differences (P < 0.05), P between 0.05 and 0.1 was considered trend
Fig. 4

Profile of immune-related gene expression in spleen post vaccination and antibody response of immunized zebrafish. A–G Relative expression of immune-related gene expression in spleen. H Serum IgM content. I Intestinal IgZ content. Data represent the means (± SD) of six replicates of each group, groups with different letters indicate significant differences (P < 0.05). P between 0.05 and 0.1 was considered trend
Fig. 5

Vaccination improved intestinal microbiota. The relative bacterial abundance at the phylum (A) and genus level (B). C PCoA of the intestinal microbiota on OTU level. D The ratio of (Firmicutes + Fusobacteria + Bacteroidetes)/Proteobacteria. Groups with different letters indicate significant differences (P < 0.05)
Fig. 6

The survival curves of zebrafish after challenged by A. veronii
Fig. 7

Gene expression of germ-free zebrafish. Gene expression induced by directly immersed germ-free zebrafish with GS115 or GS115-NTaer strain (A) and by intestinal microbiota caused by control, GS115, or GS115-NTaer vaccine diet (B). Groups with different letters indicate significant differences (P < 0.05)
Fig. 8

Survival rate of GF zebrafish. A Survival curve of GF zebrafish which directly immersed with GS115 or GS115-NTaer strain after challenged with A. veronii, the untreated group was used as control. B Survival curve of germ-free zebrafish which immersed with intestinal microbiota induced by control, GS115, or GS115-NTaer vaccine diet after challenged with A. veronii. Groups with different letters indicate significant differences (P < 0.05), P between 0.05 and 0.1 was considered trend
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Mar Life Sci Technol
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