Intestinal microbiota has become an integral component of the fish, and plays a key role in hostmetabolism, immunity and health maintenance. However, information on the immune responses aftervaccine administration in relation to the intestinal microbiota is absent in fish. The present study focusedon the effect ofa new recombinant Aeromonas hydrophilu vaccine (Aera) by using a novel functionalized,single»walled carbon nanotubes (SWCNTs) as a deliveiy vehicle on the intestinal inicrobiota of grass carp(Ctennpharyngodon idellaj through the bath immunization, and further explored the immunologicalresponses in intestine, kidney and spleen. By performing deep sequencing, a total of 81,979 valid readsand 609 OTUs obtained from 4 intestine samples were analyzed. We detected I41 genuses, most of whichbelonged to Firmicutes, Fusobacteria and Proteobacteria. Of note, the quantity of Aeromonas in libraryAera (after 6 h Aera vaccine pretreatment, fish were transferred to tanks without SWCNTs-Aera for 28 d)and Aera-GD (6 h Aera vaccine pretreated prior to the group injected by A. Iiydrophiluj was declined 6.5%and 14.6% compared with the control, respectively. Moreover. the expression of seven immune»relatedgenes (IFN-I, TNF-<1, CRP, IL-8, lgM, MHC I and CD8m) in the intestine, kidney and spleen of Aeratreated fish was significantly enhanced, which indicated that a better tissue immune response in grasscarp was induced by the SWCNTs-Aera vaccine. Therefore, a new recombinant SWCNT-Aera vaccine mayrepresent potentially efficient and immunological role in grass carp intestine to resist A. hydropliilainfection.d
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