Therapeutic effects of fumaric acid on proteomic expression and gut microbiota composition in Pacific white shrimp (Penaeus vannamei) infected with Ecytonucleospora hepatopenaei (EHP).

Recently, microsporidiosis caused by a microsporidian [Ecytonucleospora (Enterocytozoon) hepatopenaei, EHP] has been found to seriously impact the global shrimp industry. The aim of this study was to evaluate the therapeutic effects of fumaric acid (FA) in EHP-infected Pacific white shrimp (Penaeus vannamei). In the first 2 groups, non-EHP-infected shrimp were fed FA-supplemented (10 g/kg diet) or normal feed (CM+ and CM-, respectively). The other 2 groups of EHP-infected shrimp were also fed FA-supplemented or normal feed (EM+ and EM-, respectively). All the experimental groups were fed for 7 days, and the hepatopancreas and intestine of the shrimp were sampled at 0, 1, 3 and 7 days after application (DAAs). The copy number of EHP in the hepatopancreas of the EM+ shrimp was significantly lower than that in the hepatopancreas of the EM- shrimp at 3 and 7 DAAs (P<0.01). Histopathological investigation revealed that the hepatopancreas of EM+ shrimp began healing from microsporidiosis at 3 DAA and had almost completely recovered at 7 DAA. Proteomic analysis also revealed that the levels of immune-related proteins, such as β-1,3-glucan-binding proteins, the tumor suppressor TP53, and protein disulfide isomerase A3, were elevated in the hepatopancreas of the CM+ shrimp compared with those in the control shrimp. Microbiome analyses from both LC‒MS/MS data and next-generation sequencing (NGS) of the shrimp intestine revealed that FA supplementation strongly affected the bacterial community in the shrimp gut. Based on the results from this study in the hepatopancreas of shrimp fed a diet of 10 g/kg FA for 7 days, FA strongly affected EHP proliferation; simultaneously, it increased the levels of several key molecules involved in oxidative stress, cellular stress and pattern recognition without harmful negative side effects; and effectively influenced the gut microbiota. This research is the first to show the effectiveness of FA in promoting shrimp health in the context of microsporidiosis in Pacific white shrimp and could be further applied in the global shrimp aquaculture industry.