Antiviral immunity in fish is not well understood. In mammals, Toll-like receptor (TLR) 3 is involved in double-stranded RNA recognition and host immune response activation. Here, we report the first identification of a rainbow trout TLR3 ortholog (rtTLR3), its genomic structure, and mRNA regulation. Six exons and five introns were identified from bacterial artificial chromosome (BAC) and expressed sequence tag (EST) sequencing, and this genomic organization is similar to mammalian and fish TLR3 genes. The putative 913 amino acid protein has a Toll/interleukin (IL)-1R (TIR) domain, a transmembrane domain, and leucine-rich repeats. In healthy trout, rtTLR3 is highly expressed in the liver, pyloric ceca, intestine, spleen, and anterior and trunk kidney tissues. To investigate whether rtTLR3 is involved in antiviral immunity, transcriptional regulation in vivo was examined by quantitative real-time polymerase chain reaction (PCR) after poly inosinic:cytidylic (I:C) and infectious hematopoietic necrosis virus (IHNV) treatments. TLR3 mRNA expression peaked 1 day after poly (I:C) injection of live animals, while the peak of gene expression after live IHNV challenge was observed on day 3. In vitro stimulation of rainbow trout anterior kidney leukocytes with poly (I:C) also enhanced rtTLR3 expression. Up-regulation was specific to viral challenge as there was no significant up-regulation of rtTLR3 mRNA levels in the spleen and a modest down-regulation in the anterior kidney after bath challenge with a gram-negative bacterial trout pathogen, Yersinia ruckeri. The sequence conservation of trout TLR3 and mRNA regulation after poly (I:C) or RNA virus exposures strongly suggest a role for trout TLR3 in antiviral immunity.
Background: Comparative genomics, through the integration of genetic maps from species of interest with whole genome sequences of other species, will facilitate the identification of genes affecting phenotypes of interest. The development of microsatellite markers from expressed sequence tags will serve to increase marker densities on current salmonid genetic maps and initiate in silico comparative maps with species whose genomes have been fully sequenced.
Abajo: se repite los tres en el resumen. This work studies the effect of high-level fish meal replacement with insect meal: YW meal (obtained from Tenebrio larvae fed a broiler diet), BSF meal (from hermetia larvae fed broilers diet), BSFm meal (obtained from hermetia larvae fed discard fish) on growth performance nutritive indices and in vitro digestibility of Dicentrarchus labrax juvenile. Three different insect meals were used: BSF meal from hermetia larvae fed broilers diet; BSF improve (BSFm) obtained from hermetia larvae fed discarded fish; YW meal obtained from the larvae of Tenebrio fed a broiler diet. Five diets were used, a control (C) diet and four experimental diets by replacing fishmeal with insect meal from BSF at 30% and 50% (BSF30 and BSF50) substitutions, BSFm at 50% substitution (BSF50 m) and YM at 50% substitution (YW50). Nutritional and growth indices worsened by including insect meal, especially for hermetia meal at 50% substitution, BSF50 and BSF50 m. The internal organs’ weight reflected the growth of the fish fed each experimental diet. No differences were found in fillet composition. Nevertheless, under our experimental condition, YW replacement obtained better results than both BSF diets.
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