Two routes of infection with Photobacterium damselae subsp. piscicida are effective in the modulation of the transcription of immune related genes in Solea senegalensis

Abstract

The marine fish pathogen Photobacterium damselae subsp. piscicida (Phdp) is responsible for important disease outbreaks affecting cultured fish species including the flatfish Solea senegalensis. In the present work, transcription of iron metabolism related genes (TF, FERR-M, HP-1 and HAMP-1) as well as innate immune system components such as complement proteins (C3 and C7), lysozyme (LYS-G), TNF family (TNFα, TRAF-3), NCCRP-1 and heat shock protein encoding genes (HSP70, HSP90AA, HSP90AB and GP96) has been determined in the liver and kidney of S. senegalensis specimens after Phdp infection. Intraperitoneal injection (IP) and immersion (IM) routes have been used for infection. Fish developed specific antibodies in both cases, higher levels being detected in IP infected specimens. Both infection routes resulted in increased relative transcript levels of FERR-M, HP-1 and HAMP-1 genes and TF decreased relative transcription, conducting to lower iron availability for the pathogen. This response can be considered as a strategy to limit iron availability for Phdp, a pathogen capable to obtain iron from transferrin. Relative transcription of genes encoding lysozyme and complement factors C3 and C7 were also increased regardless the infection route; the liver was the main organ involved in the initial stages and the kidney in later stages of the infection. TNFα and TRAF-3 relative gene transcription increased 24h post-infection. TRAF-3 gene induction was detected 30 d post-infection, whilst no changes in TNFα were observed 72h or 30 d post-infection. NCCRP-1 changes were observed after IP infection in the liver and kidney; however, IM infection resulted only in slight changes in the kidney of infected fish. This different response observed maybe related to a lower number of invaded cells by the pathogen. Finally, changes in HSP90AB and GP96 have been detected after infection by both routes. Different late modulation has been observed in assayed genes depending on the route of infection. Thus, only LYS-G, TF, NCCRP-1, GP96 and HSP90AB gene transcription was modulated 30 d post-infection in the kidney of IM infected specimens; however, IP infected fish showed modulation in a higher number of genes both in liver and kidney tissues. The implications of these responses in resistance to infection by Phdp need to be elucidated.