Effects of dietary arachidonic acid (ARA) on immunity, growth and fatty acids of Apostichopus japonicus.

As an important aquaculture species, improving the immunity of cultured Apostichopus japonicus (A. japonicus) is vital for its health in aquaculture farming. It has been shown that ARA is an important metabolite for A. japonicus infected by Vibrio splendidus. In this study, we aimed to determine the effects of dietary exogenous ARA on healthy sea cucumber cultures, including assessments of immunity, growth, and fatty acid content. Five experimental diets containing 0.01%, 0.29%, 0.46%, 0.70%, and 1.09% ARA were tested. The specific growth rate (SGR) of sea cucumbers did not be significantly affected by exogenous ARA diet groups. The results showed that dietary ARA between 0.49 and 1.09% notably improved the survival rate of sea cucumbers infected by Vibrio splendidus compared with the control group without exogenous ARA. The results also showed the effects of dietary ARA on immune-related genes, enzymes, and oxidation indices; most of the exogenous ARA significantly upregulated the mRNA expression of the genes NFκB, TLR, TLR3, TRAF6, Toll, and MyD88. The activities of ACP, AKP, and lysozyme increased in the 0.49-1.09% ARA groups, especially the dietary 0.49% ARA group. The SOD1 and NOS activities were enhanced by dietary ARA between 0.29 and 0.70%. Compared with the control, the MDA content increased, but the 0.49% ARA-diet group had a lower MDA content. Based on these data, 0.49-0.70% ARA significantly enhanced immunity in cultured A. japonicus. Exogenous 0.49% and 0.70% ARA also elevated the ARA, total PUFA and n-6 PUFA in the body walls. In conclusion, the appropriate exogenous ARA (0.49%-0.70%) in diets could improve immunity and fatty acid content considerably. The results provide basic evidence that ARA can serve a useful immune enhancer for A. japonicus aquaculture.

The arachidonic acid and its metabolism pathway play important roles for Apostichopus japonicus infected by Vibrio splendens.

Improving the immune ability and guiding healthy culture for sea cucumber by purposefully screening the significant differential metabolites when Apostichopus japonicus (A. japonicus) is infected by pathogens is important. In this study, 35 types of significant differential metabolites appeared when A. japonicus were infected by Vibrio splendens (VSI group) compared with the control A. japonicus group (CK group) by using liquid chromatography-mass spectrometry (LC-MS/MS)-based untargeted metabolomics. Based on that finding, the 10 types of key metabolic pathways were analyzed by MetPA. The "arachidonic acid (ARA) metabolism" pathway, which was screened by three elevated biomarkers: ARA, prostaglandin F2α and 2-arachidonoyl glycerol, had an important impact on immune stress in A. japonicus. Due to the similar changes in several metabolites in its metabolic pathway, the ARA metabolic pathway was selected for further study. The activities of ACP, AKP and lysozyme, which are important innate immune-related enzymes, the survival rates of A. japonicus infected with V. splendidus and the relative content of ARA in the body wall detected by GC-MS were all upregulated significantly by exogenous daily 0.60% and 1.09% ARA consumption over a short period of approximately 7 days. These results demonstrated that ARA and its metabolic pathway indeed played important roles in the immunity of A. japonicus infected by the pathogen. The findings also provide novel insights for the effects of metabolites in A. japonicum healthy culture.

High production of d-psicose from d-fructose by immobilized whole recombinant Bacillus subtilis cells expressing d-psicose 3-epimerase from Agrobacterium tumefaciens.

d-Psicose 3-epimerase (DPEase) can catalyze the isomerization of d-fructose to be rare sugar d-psicose, which has wide application prospects in the food and medical fields. In this study, the DPEase gene from Agrobacterium tumefaciens was constructed into plasmid pMA5, and was successfully expressed in the host Bacillus subtilis WB600 (B. subtilis). After optimization of the fermentation conditions, whole recombinant B. subtilis WB600/pMA5-At-DEPase(O) cells produced d-psicose from d-fructose with a conversion rate of 29.01 ± 0.19%, which could be used for the efficient synthesis of d-psicose. To further improve the whole recombinant B. subtilis application, B. subtilis cells were immobilized onto a gel bead biocatalyst by Ca-alginate. After optimization of the biotransformation conditions, the conversion rate of the immobilized biocatalyst reached 20.74 ± 0.39%, which was lower than the free cells. However, the results showed that the immobilized biocatalyst had higher thermal/pH stability and storability, and the gel beads could be recycled for at least six batches. The results showed that the amount of d-psicose generated reached 32.83 ± 2.56 g/L with the immobilized biocatalyst after six times biotransformation, whereas the free cells produced only approximately 10.44 ± 0.07 g/L. The results showed that immobilized recombinant B. subtilis cells are promising to use for the efficient synthesis of d-psicose.