Effects of dietary vitamin D3 supplementation on the growth performance, tissue Ca and P concentrations, antioxidant capacity, immune response and lipid metabolism in Litopenaeus vannamei larvae.

An 8-week feeding trial was conducted to investigate the effects of dietary vitamin D3 supplementation on the growth performance, tissue Ca and P concentrations, antioxidant capacity, immune response and lipid metabolism in Litopenaeus vannamei larvae. A total of 720 shrimp (initial weight 0·50 ± 0·01 g) were randomly distributed into six treatments, each of which had three duplicates of forty shrimp per duplicate. Six isonitrogenous and isolipidic diets were formulated to contain graded vitamin D3 (0·18, 0·23, 0·27, 0·48, 0·57 and 0·98 mg/kg of vitamin D3, measured) supplementation levels. The results revealed that L. vannamei fed diet containing 0·48 mg/kg of vitamin D3 achieved the best growth performance. Compared with the control group, supplementing 0·48 mg/kg of vitamin D3 significantly increased (P < 0·05) the activities of catalase, total antioxidative capacity, alkaline phosphatase and acid phosphatase in serum and hepatopancreas. Expression levels of antioxidant and immune-related genes were synchronously increased (P < 0·05). Carapace P and Ca concentrations were increased (P < 0·05) with the increased vitamin D3 supplementation levels. Further analysis of lipid metabolism-related genes expression showed that shrimp fed 0·48 mg of vitamin D3 per kg diet showed the highest value in the expression of lipid synthesis-related genes, while shrimp fed 0·98 mg of vitamin D3 per kg diet showed the highest value in the expression of lipolysis-related genes. In conclusion, the results of present study indicated that dietary supplementation of 0·48 mg/kg of vitamin D3 could increase Ca and P concentrations, improve antioxidant capacity and immune response, and influence lipid metabolism in L. vannamei.

Excess iron supplementation induced hepatopancreas lipolysis, destroyed intestinal function in Pacific white shrimp Litopenaeus vannamei.

So far, the adverse effects of excess Fe in shrimp have been ignored for years as it was thought that extra Fe supplementation was not needed in the practical diets. Nowadays, Fe concentration in commercial shrimp feed from feed enterprises could be around 301.34-545.5 mg/kg, which is mainly due to the fish meal containing up to 1500 mg/kg Fe. Therefore, the purpose of this experiment was to investigate the effects of Fe supplementation on the growth performance, tissue Fe deposition, hepatopancreas lipid metabolism, intestinal function in L. vannamei. The results showed that although growth performance was not influenced by the dietary Fe supplementation, excess Fe supplementation (955.00 mg/kg) significantly increased hepatopancreas Fe deposition and induced lipolysis. Moreover, excess Fe supplementation impaired intestinal immune function and disrupted microbiota homeostasis. These findings might provide partial theoretical evidence for the effect of dietary Fe supplementation on physiological metabolism in L. vannamei.

Dietary vitamin K3 activates mitophagy, improves antioxidant capacity, immunity and affects glucose metabolism in Litopenaeus vannamei.

An 8-week feeding experiment was conducted to appraise the influence of dietary vitamin K3 on the growth performance, antioxidant capacities, immune responses, mitophagy and glucose metabolism in Litopenaeus vannamei. Six diets containing graded dietary vitamin K3 (0.40(control), 9.97, 20.29, 39.06, 79.81 and 156.02 mg kg-1 of vitamin K3, respectively) levels were formulated. A total of 900 shrimp with 0.90 g initial weight were randomly assigned to six diets with three replications. Our results revealed that diets supplemented with 9.97-156.02 mg kg-1 vitamin K3 didn't affect the growth performance in L. vannamei. In general, compared with the control group, 39.06 mg kg-1 vitamin K3 group significantly increased (P < 0.05) the total antioxidative capacity, and the activities of catalase, glutathione, nitric oxide synthase, alkaline phosphatase and acid phosphatase in serum and hepatopancreas. 39.06 mg kg-1 vitamin K3 group significantly decreased (P < 0.05) the malondialdehyde in serum and hepatopancreas. The mRNA levels of antioxidant and immune related genes were increased synchronously (P < 0.05). In addition, 39.06 mg kg-1 vitamin K3 group increased glycogen content and levels of mitophagy (pink1, ampkα, parkin, lc3, atg13, atg12) genes. Expression levels of glucose transport related gene (glut1), glycolysis related genes (hk, pfk), glycogen synthesis related genes (gsk-3ß, gys), insulin-like peptides (ILPs)/AKT/PI3K pathway related genes (insr, irsl, akt, pi3k, pdpk1) were increased in the hepatopancreas of 39.06 mg kg-1 vitamin K3 group. In conclusion, the present results indicated that although dietary supplementing vitamin K3 had no influence on the growth performance, 39.06 mg kg-1 vitamin K3 could activate ampkα/pink1/parkin mediated mitophagy, improve antioxidant capacity and immune response. Moreover, vitamin K3 could trigger ILPs/AKT/PI3K signaling pathways and influence glucose metabolism in L. vannamei. This finding would help to advance the field of vitamin K3 nutrition and guide the development of future crustacean feeds.

Litopenaeus vannamei BMAL1 Is a Critical Mediator Regulating the Expression of Glucose Transporters and Can Be Suppressed by Constant Darkness.

Aryl hydrocarbon receptor nuclear translocator-like protein 1 (BMAL1) is a core circadian transcription factor that controls the 24-h cycle of physiological processes. In shrimp, the role of BMAL1 in the regulating glucose metabolism remains unclear. Firstly, we observed that the daily profile of BMAL1, GLUT1 and SGLT1 expression were synchronized in the intestine and the hepatopancreas of Litopenaeus vannamei. Then we examined the effects of BMAL1 on the gene expression of glucose transporter type 1 (SGLT1) and sodium-glucose cotransporter 1 (GLUT1) in vivo and in vitro. BMAL1 in L. vannamei shares 70.91-96.35% of sequence identities with other shrimp species and possesses the conserved helix-loop-helix domain and polyadenylation site domain. The in vitro dual-luciferase reporter assay and in vivo RNA interference experiment demonstrated that BMAL1 exerted a positive regulation effect on the expression of glucose transporters in L. vannamei. Moreover, we conducted an eight-week treatment to investigate whether light/dark cycle change would influence growth performance, and gene expression of BMAL1, GLUT1 and SGLT1 in L. vannamei. Our result showed that compared with natural light treatment, constant darkness (24-h darkness) significantly decreased (p < 0.05) serum glucose concentration, and suppressed (p < 0.05) the gene expression of BMAL1, GLUT1 and SGLT1 in the hepatopancreas and the intestine. Growth performance and survival rate were also decreased (p < 0.05) by constant darkness treatment. Our result identified BMAL1 as a critical mediator regulating the expression of glucose transporters, which could be suppressed by constant darkness in L. vannamei. It would be quite interesting to explore the mechanism of dark/light cycles on glucose transport and metabolism in L. vannamei, which might provide a feeding strategy for improving carbohydrate utilization in the future.

Transcriptome Analysis of the Hepatopancreas in the Litopenaeus vannamei Responding to the Lead Stress.

Lead (Pb) is one of the most hazardous pollutants and toxic heavy metal in marine environment. The molecular mechanisms of Pb toxicity in aquatic organism are not well understood. In this study, hepatopancreas transcriptome of Litopenaeus vannamei (L. vannamei) was characterized by a comparison between control and Pb exposure samples using RNA-Seq approach. Hepatopancreas morphology of L. vannamei was also assessed. The result reveals that compared with the control group, an increase in the number of B cells was observed following Pb exposure in L. vannamei. Transcriptome data showed that a total of 1593 genes were recognized to be differentially expressed including 1278 up-regulated and 315 down-regulated genes. These genes were mainly associated with energy metabolism, cell apoptosis, exogenous microbial infection, cell junction, and cell adhesion. Fifteen ribosomal protein genes (RPS3, RPS13, RPSA, RPL11, RPS2, RPL8, RPS23, RPL3, RPL5, RPS6, RPS4X, RPS18, RPL19, RPL9, RPL6) were identified as the common hubs of protein-protein interaction (PPI) networks, as well as part of modules of the PPI network. Besides ribosomal protein, we identified differential expression genes (DEGs) including GAPDH, EEF1A1, HSPA8, UBC, and EEF1G as the common hubs of PPI networks. These findings may have important implications for understanding the adverse biological effects of Pb and its toxic mechanisms, as yet not clearly defined, and provide potential biomarkers of Pb exposure in hepatopancreas of L. vannamei, which might be useful for monitoring aquatic environments and assessing the health of the marine ecosystem.

Vibrio parahaemolyticus Infection Influenced Trace Element Homeostasis, Impaired Antioxidant Function, and Induced Inflammation Response in Litopenaeus vannamei.

Vibrio parahaemolyticus (V. parahaemolyticus) caused huge diseases and economic losses in shrimp aquaculture. Understanding the infection mechanism might help develop new strategies for controlling pathogen outbreak. Redistribution of trace element homeostasis, accompanied by impairment of antioxidant status and immune response, was observed during various infections. Accordingly, we hypothesized that V. parahaemolyticus infection might influence trace element homeostasis, impair antioxidant function, and induce inflammation response in shrimp. In the present study, the aim of this study was to investigate the influence of V. parahaemolyticus infection on trace element homeostasis, antioxidant status, and inflammation response in Litopenaeus vannamei (L. vannamei). The results showed that compared with the control group, V. parahaemolyticus infection significantly increased (P < 0.05) intestinal V. parahaemolyticus number, serum copper (Cu) concentration at 24, 48, and 72 h and significantly increased (P < 0.05) serum zinc (Zn), iron (Fe), and manganese (Mn) concentrations at 24 h but decreased (P < 0.05) at 72 h. The intestinal gene expressions of metal transporters ZIP13, CTR1, and MT1 were significantly decreased at 24, 48, and 72 h, and DMT1 was significantly decreased at 48 h and 72 h in the infection group. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were suppressed at 48 h and 72 h, and the malondialdehyde (MDA) content was increased at 24, 48, and 72 h in the infection group; the pro-inflammatory genes including necrosis factor-α (TNF-α), lipopolysaccharide-induced TNF-α factor (LITAF), and Ras-related protein Rab6A (RAB6A) were significantly upregulated at 48 and 72 h in the infection group. These results suggest that V. parahaemolyticus infection influenced trace element homeostasis, impaired antioxidant function, and induced inflammation response in L. vannamei, which might help understand the infection mechanism. The results provide a better understanding of the L. vannamei and V. parahaemolyticus interactions and may deliver the basis for further research in preventing the bacterial diseases.

New insight into the molecular basis of chromium exposure of Litopenaeus vannamei by transcriptome analysis.

Heavy metal pollution arising from agricultural and industrial activities poses a significant threat to the aquatic environment, especially the increasing levels of chromium (Cr) that is exacerbating marine pollution. Given the economic importance of the Pacific white shrimp Litopenaeus vannamei (L. vannamei), understanding the impact of marine Cr pollution is deemed to be significant. In this study, we used the transcriptome sequencing (RNA-seq) technique to characterize the molecular mechanism of Cr exposure in L. vannamei. Gene ontology enrichment analysis showed substrate-specific and ion transport-related functions were mainly influenced by Cr exposure. We further identified genes involved in protein digestion and absorption (PEPT1, BAT1, MDU1), chemical carcinogenesis (GST and UGTs), ABC transporters (ABCC2), apoptosis (CAPN1, CASP10, PARP), implying the potentially Cr disintoxication mechanisms in L. vannamei. Genes within pancreatic secretion (ALT, LDH), lysosome (CTSL and HEXB), and peroxisome (ACOX1, ECI2, NUDT12) pathways implied the potentially Cr toxicity mechanisms in L. vannamei.