Gamma-aminobutyric acid enhances hypoxia tolerance of juvenile Chinese mitten crab (Eriocheir sinensis) by regulating respiratory metabolism and alleviating neural excitotoxicity.

With climate change and intensive aquaculture development, environmental hypoxia in aquaculture water has become a common challenge for many aquatic species. Therefore, it is crucial to improve the hypoxic tolerance of animals through nutritional strategies. This study explored the positive role of dietary gamma-aminobutyric acid (GABA) supplementation in enhancing hypoxia tolerance of juvenile Eriocheir sinensis through respiratory regulation and alleviation of hypoxia-induced neural excitotoxicity. Acute hypoxia stress significantly up-regulated the mRNA expression level of hypoxia-inducible factor 1α, oxygen consumption rate and anaerobic respiratory metabolism-related enzyme activities. On the other hand, aerobic respiratory metabolism-related enzyme activities were significantly decreased. However, dietary GABA supplementation remodeled the respiratory metabolism pattern of juvenile crabs exposed to hypoxia stress. In addition, acute hypoxic stress significantly increased the contents of free glutamate and GABA in the nervous tissue. The expression levels of N-Methyl-d-aspartate-related receptor genes and calcium-dependent degradation enzyme-related genes were significantly up-regulated. Similarly, neuronal apoptosis rates, expression levels of apoptosis-related genes, and vesicular glutamate transporter genes were also significantly increased. The high-affinity neuronal glutamate transporter decreased significantly in the crabs exposed to hypoxia stress. However, dietary GABA supplementation could effectively prevent acute hypoxia stress-induced neural excitotoxicity. Furthermore, dietary GABA could significantly improve the redox status in vivo exposed to hypoxia stress. In conclusion, acute hypoxia stress can affect respiratory metabolism and redox state and induce neural excitotoxicity in juvenile E. sinensis. GABA supplementation could improve hypoxia tolerance through multiple physiological regulation pathways.

Myo-inositol improves growth performance and regulates lipid metabolism of juvenile Chinese mitten crab (Eriocheir sinensis) fed different percentage of lipid.

This study evaluated the effects of dietary myo-inositol (MI) on growth performance, antioxidant status and lipid metabolism of juvenile Chinese mitten crab (Eriocheir sinensis) fed different percentage of lipid. Crabs (4·58 (sem 0·05) g) were fed four diets including a normal lipid diet (N, containing 7 % lipid and 0 mg/kg MI), N with MI supplementation (N + MI, containing 7 % lipid and 1600 mg/kg MI), a high lipid diet (H, containing 13 % lipid and 0 mg/kg MI) and H with MI supplementation (H + MI, containing 13 % lipid and 1600 mg/kg MI) for 8 weeks. The H + MI group showed higher weight gain and specific growth rate than those in the H group. The dietary MI could improve the lipid accumulations in the whole body, hepatopancreas and muscle as a result of feeding on the high dietary lipid (13 %) in crabs. Besides, the crabs fed the H + MI diets increased the activities of antioxidant enzymes but reduced the malondialdehyde content in hepatopancreas compared with those fed the H diets. Moreover, dietary MI enhanced the expression of genes involved in lipid oxidation and exportation, yet reduced lipid absorption and synthesis genes expression in the hepatopancreas of crabs fed the H diet, which might be related to the activation of inositol 1,4,5-trisphosphate receptor (IP3R)/calmodulin-dependent protein kinase kinase-ß (CaMKKß)/adenosine 5'-monophosphate-activated protein kinase (AMPK) signalling pathway. This study demonstrates that MI could increase lipid utilisation and reduce lipid deposition in the hepatopancreas of E. sinensis fed a high lipid diet through IP3R/CaMKKß/AMPK activation. This work provides new insights into the function of MI in the diet of crustaceans.

Impact of Dietary Vitamin D3 Supplementation on Growth, Molting, Antioxidant Capability, and Immunity of Juvenile Chinese Mitten Crabs (Eriocheir sinensis) by Metabolites and Vitamin D Receptor.

Vitamin D3 (vit-D3), as an indispensable and fat-soluble nutrient, is associated with skeletal mineralization and health in mammals. However, such associations have not been well studied in economically important crustaceans. Six levels of vit-D3 with isonitrogenous and isolipidic diets were used to feed Eriocheir sinensis. The range of optimal vit-D3 requirements is 5685.43-10,000 IU/kg based on growth. The crabs fed 9000 IU/kg vit-D3 showed the best growth performance. This vit-D3 dose significantly increased antioxidant capacity in the hepatopancreas and intestine and was optimal for molting and innate immunity via quantitative polymerase chain reaction analysis. Transcriptomics analyses indicate that vit-D3 could alter protein processing in the endoplasmic reticulum, steroid biosynthesis, and antigen processing and presentation. As shown by the enzyme-linked immunosorbent assay, vit-D3 could improve vitamin D receptor, retinoic acid receptor, and C-type lectins concentrations. The 1α,25-dihydroxy vit-D3 content in serum was significantly higher in 3000-9000 IU/kg vit-D3. The study suggests that dietary vit-D3 and its metabolites can regulate molting and innate immunity in crabs.