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The Effects of Immunostimulation Through Dietary Manipulation in the Rainbow Trout; Evaluation of Mucosal Immunity

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Abstract

Immunostimulant-containing diets are commonly used in aquaculture to enhance the resistance of cultured fish to disease and stress. Although widespread in use, there have been conflicting results published, and surprisingly little is known about the regulation of immune response-related genes in tissues key to mucosal immunity induced by immunostimulant dietary feeding. Using a salmonid-specific microarray platform enriched with immune-related genes and in situ hybridization, we investigated dietary acclimation in two organs relevant to mucosal immunity, the gills and the intestine, in the rainbow trout (Oncorhynchus mykiss). Immunostimulant diets significantly changed gene expression profiles and gene distribution in a tissue-specific manner: genes and functional Gene Ontology categories involved in immunity were differently expressed at portals of entry where significant changes in genes and functional groups related to remodeling processes and antigen presentation were observed. Furthermore, genes involved in chemotaxis, cell differentiation, antigen-presenting capacity and tissue remodeling were localized in both organs.

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Acknowledgments

This work was supported by Plan Nacional AGL2005-12134 from the Ministerio de Educación y Ciencia, España, and the pre-doctoral fellowship AP20047208 from the Ministerio de Educación y Ciencia.

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Authors and Affiliations

  1. Unitat de Fisiologia Animal, Departament de Biologia Cellular, Fisiologia i d’Immunologia, Facultat de Ciencies, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain

    Carmen Doñate, Joan Carles Balasch, Agnes Callol, Lluis Tort & Simon MacKenzie

  2. INRA, SCRIBE UR1037, IFR 140, Campus de Beaulieu, 35000, Rennes, France

    Julien Bobe

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  1. Carmen Doñate
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  2. Joan Carles Balasch
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  3. Agnes Callol
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  4. Julien Bobe
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  5. Lluis Tort
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  6. Simon MacKenzie
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Correspondence to Simon MacKenzie.

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Additional file 1

Total number of differentially expressed genes. Differentially expressed genes in fish fed with immunostimulant diet (t Student, p < 0.01). a Gills. b Intestine (FI fold induction, FR fold repression) (GIF 17 kb)

High resolution image file (TIFF 141 kb)

Additional file 2

Primers used for Q-PCR (DOC 38 kb)

Additional file 3

Functional Gene Ontology categories. Gene Ontology analysis in gills (a) and intestine (b) that discriminate between control fish and fish fed with immunostimulant diet for 4 weeks. Functional categories compared pairwise by the sums of ranks of differentially expressed genes (p < 0.01) Significance determined with Student’s t test (p < 0.05; DOC 63 kb)

Additional file 4

Differentially expressed genes in gills. Differentially expressed genes in the gills of fish fed with immunostimulant diet. Differential expression was analyzed with Student’s t test (p < 0.01). Values are expressed as fold difference, FD. Harvester links are provided (DOC 179 kb)

Additional file 5

Differentially expressed genes in intestine. Differentially expressed genes in the intestine of fish fed with immunostimulant diet. Differential expression was analyzed with Student’s t test (p < 0.01). Values are expressed as fold difference, FD. Harvester links are provided (DOC 795 kb)

Additional file 6

Representative photos of several fish at ×20 magnification illustrating the localization by in situ hybridization of mRNA expression in gills and intestine in fish fed with control diet and fish fed with immunostimulant diet. Positive reaction shows blue. a Pyrin-2 expression in gills in control fish and in fish fed with immunodiet. Positive hybridizations are localized across primary and secondary lamellae. Stronger hybridization signals were observed in epithelial cells at the extreme of the secondary lamellae (arrowheads). b MMP9-1 expression in gills fed with control diet. Positive cells are distributed across primary and secondary lamellae, more abundant in inter-lamellar cells (arrow; PL primary lamellae, SL secondary lamellae, GS gill support, asterisk: mucous cells, CS cartilaginous support, C capillary). C: MLF-1 expression in intestine of trout fed with control diet. Strong signal is present in the epithelium near the lamina propia (arrowhead) (L: lumen, LP: lamina propia, asterisk: mucous cells) (GIF 303 kb)

High resolution image file (TIFF 4.5 mb)

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Doñate, C., Balasch, J.C., Callol, A. et al. The Effects of Immunostimulation Through Dietary Manipulation in the Rainbow Trout; Evaluation of Mucosal Immunity. Mar Biotechnol 12, 88–99 (2010). https://doi.org/10.1007/s10126-009-9203-4

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