UNASSIGNED: Oxygen concentration is an essential water quality parameter for aquaculture systems. Recently, supersaturated dissolved oxygen (DO) has been widely used in aquaculture systems to prevent oxygen depletion; however, the long-term effects of supersaturated DO exposure on aquatic animals have not been studied. In this study, we examined the effects of supersaturated DO on the growth, survival, and gene expression of Pacific white shrimp (Litopenaeus vannamei).
UNASSIGNED: Specific pathogen-free shrimp with a body weight of 8.22 ± 0.03 g were randomly assigned to two groups with four replicates at a density of 15 shrimps per tank. Shrimp were cultivated in recirculating tanks containing 50 L of 15 ppt seawater in each replicate. Oxygen was supplied at 5 mg/L to the control tanks using an air microbubble generator and at 15 mg/L to the treatment tanks using a pure oxygen microbubble generator. Shrimp were fed commercial feed pellets containing 39% protein at 4% of their body weight per day for 30 days. Average daily growth (ADG) and feed conversion ratio (FCR) were determined on days 15 and 30. Shrimp molting was measured every day. Individual hemolymph samples were obtained and analyzed for total hemocyte count, differential hemocyte count, and expression of growth- and immune-related genes at the end of the experiment.
UNASSIGNED: Long-term exposure to supersaturated DO significantly affected shrimp growth. After 30 days of supersaturated DO treatment, the final weight and ADG were 14.73 ± 0.16 g and 0.22 ± 0.04, respectively. Shrimp treated with normal aeration showed significantly lower weight (12.13 ± 0.13 g) and ADG (0.13 ± 0.00) compared with the control group. FCR was 1.55 ± 0.04 in the treatment group and 2.51 ± 0.09 in the control group. Notably, the shrimp molting count was 1.55-fold higher in the supersaturated DO treatment than in the supersaturated DO treatment. The expression of growth-related genes, such as alpha-amylase, cathepsin L, and chitotriosidase, was 1.40-, 1.48-, and 1.35-fold higher, respectively, after supersaturated DO treatment. Moreover, the treatment increased the expression of anti-lipopolysaccharide factor, crustin, penaeidin3, and heat shock protein 70 genes by 1.23-, 2.07-, 4.20-, and 679.04-fold, respectively, compared to the controls.
UNASSIGNED: Supersaturated DO increased growth and ADG production and decreased FCR. Furthermore, enhanced immune-related gene expression by supersaturated DO may improve shrimp health and reduce disease risk during cultivation.

Fried shrimp are popular for their attractive organoleptic and nutritional qualities. However, consumers are more concerned about the safety of fried foods. To investigate the formation of advanced glycation end products (AGEs) in fried shrimp and provide pretreatment guidance for producing low-AGEs fried pacific white shrimp were treated with seven pretreatment methods before frying. The AGEs contents, physicochemical indicators, and their correlations in the fried shrimps\' interior, surface, and batter layer were analyzed. Results indicated that pretreatment methods influenced both Maillard and oxidation reactions by altering the basic compositions, which controlled the formation of AGEs. The highest and lowest AGEs contents were obtained in shelled shrimp with exscinded back and whole shrimp, respectively. The batter-coated treatment reduced the AGEs contents in samples but increased the oil content. Correlation analysis showed that lipid oxidation was the decisive chemical reaction to the formation of AGEs by promoting the generation of dicarbonyl compounds and their combination with free amino acids. Conclusively, the whole shrimp was suitable for producing fried shrimp with low AGEs, oil content, and desirable color.

Shrimp has been known for its delicacy, but it undergoes rapid deterioration induced by biochemical and microbiological reactions. Melanosis is a major cause of discoloration associated with consumer rejection. All ethanolic extracts from different leaves including soursop, noni, and Jik leaves were dechlorophyllized via the \"Green\" sedimentation method before being used. The inhibitory activity against polyphenoloxidase (PPO) from Pacific white shrimp (Litopeneous vannamei) and the copper-chelating properties of varying extracts were compared. Soursop leaf extract (SLE) showed higher PPO inhibitory activity and copper-chelating ability than others (p < 0.05). Based on LC-MS, aempferol-3-O-rutinoside was identified as the most abundant compound, followed by catechin and neocholorigenic acid. The efficacy of SLE at different levels (0.25-1%) for inhibiting melanosis and preserving the quality of Pacific white shrimp was evaluated during refrigerated storage at 4 °C for 12 days in comparison with that of a 1.25% sodium metabisulfite (SMS)-treated sample. SLE at a level of 1% effectively retarded melanosis and bacterial growth, in which the total viable count did not exceed the microbial limit within 12 days. In addition, 1% SLE treatment impeded autolysis, reduced protein degradation and decomposition, and minimized lipid oxidation, as witnessed by the lower increases in pH, TVB-N, and TBARS values. Sensory evaluation indicated higher likeness scores and overall acceptability for SLE-1% and SMS-1.25% shrimps than those of the control and other samples. Therefore, SLE could be used as a natural alternative that effectively lowered the melanosis and quality loss of shrimp during refrigerated storage.

An 8-week feeding trial was conducted in Pacific white shrimp (Litopenaeus vannamei) to evaluate the effects of dietary choline supplementation on choline transport and metabolism, hepatopancreas histological structure and fatty acid profile, and regulation of lipid metabolism. Six isonitrogenous and isolipidic diets were formulated to contain different choline levels of 2.91 (basal diet), 3.85, 4.67, 6.55, 10.70 and 18.90 g/kg, respectively. A total of 960 shrimp (initial weight, 1.38 ± 0.01 g) were distributed randomly into twenty-four 250-L cylindrical fiber-glass tanks, with each diet assigned randomly to 4 replicate tanks. The results indicated that dietary choline significantly promoted the deposition of choline, betaine and carnitine (P < 0.05). The diameters and areas of R cells, total lipid and triglyceride contents in hepatopancreas, and triglyceride and non-esterified fatty acid contents in hemolymph were negatively correlated with dietary choline level. The contents of functional fatty acids in hepatopancreas, the activity of acetyl-CoA carboxylase (Acc), and the mRNA expression of fas, srebp and acc were highest in shrimp fed the diet containing 4.67 g/kg choline, and significantly higher than those fed the diet containing 2.91 g/kg, the lowest level of choline (P < 0.05). The number of R cells, content of very low-density lipoprotein (VLDL), activities of carnitine palmitoyl-transferase (Cpt1), lipoprotein lipase and hepatic lipase, and the mRNA expression levels of cpt1, fabp, fatp, ldlr, and ampk in hepatopancreas increased significantly as dietary choline increased (P < 0.05). In addition, hepatopancreas mRNA expression levels of ctl1, ctl2, oct1, badh, bhmt, ck, cept, and cct were generally up-regulated as dietary choline level increased (P < 0.01). In conclusion, dietary choline promoted the deposition of choline and its metabolites by up-regulating genes related to choline transport and metabolism. Moreover, appropriate dietary choline level promoted the development of hepatopancreas R cells and maintained the normal accumulation of lipids required for development, while high dietary choline not only promoted hepatopancreas lipid export by enhancing VLDL synthesis, but also promoted fatty acid β-oxidation and inhibited de novo fatty acid synthesis by activating the Ampk/Srebp signaling pathway. These findings provided further insight and understanding of the mechanisms by which dietary choline regulated lipid metabolism in L. vannamei.

Vibriosis is caused by some pathogenic Vibrio and produces significant mortality in Pacific white shrimp Penaeus (Litopenaeus) vannamei larvae in commercial hatcheries. Acute hepatopancreatic necrosis disease (AHPND) is an emerging vibriosis affecting shrimp-producing countries worldwide. Zoea 2 syndrome is another type of vibriosis that affects the early stages of P. vannamei larvae. Although the pathogenesis of AHPND and zoea 2 syndrome is well known, there is scarce information about microbial composition and biomarkers of P.vannamei larvae affected by AHPND, and there is no study of the microbiome of larvae affected by zoea 2 syndrome. In this work, we characterized the microbiome of P. vannamei larvae collected from 12 commercial hatchery tanks by high-throughput sequencing. Seven tanks were affected by AHPND, and five tanks were affected by zoea 2 syndrome. Subsequently, all samples were selected for sequencing of the V3-V4 region of the16S rRNA gene. Similarity analysis using the beta diversity index revealed significant differences in the larval bacterial communities between disease conditions, particularly when Vibrio was analyzed. Linear discriminant analysis with effect size determined specific microbial signatures for AHPND and zoea 2 syndrome. Sneathiella, Cyclobacterium, Haliea, Lewinella, among other genera, were abundant in AHPND-affected larvae. Meanwhile, Vibrio, Spongiimonas, Meridianimaribacter, Tenacibaculum, among other genera, were significantly abundant in larvae affected by zoea 2 syndrome. The bacterial network at the phylum level for larvae collected from tanks affected by AHPND showed greater complexity and connectivity than in samples collected from tanks affected by zoea 2 syndrome. The bacterial connections inter Vibrio genera were higher in larvae from tanks affected by zoea 2 syndrome, also presenting other connections between the genera Vibrio and Catenococcus. The identification of specific biomarkers found in this study could be useful for understanding the microbial dynamics during different types of vibriosis.

To characterize the cold tolerance mechanism of the Pacific white shrimp (Litopenaeus vannamei), we performed single-cell RNA sequencing (scRNA-seq) of ∼5185 hepatopancreas cells from cold-tolerant (Lv-T) and common (Lv-C) L. vannamei at preferred and low temperatures (28°C and 10°C, respectively). The cells fell into 10 clusters and 4 cell types: embryonic, resorptive, blister-like, and fibrillar. We identified differentially expressed genes between Lv-T and Lv-C, which were mainly associated with the terms \"immune system,\" \"cytoskeleton,\" \"antioxidant system,\" \"digestive enzyme,\" and \"detoxification,\" as well as the pathways \"metabolic pathways of oxidative phosphorylation,\" \"metabolism of xenobiotics by cytochrome P450,\" \"chemical carcinogenesis,\" \"drug metabolism-cytochrome P450,\" and \"fatty acid metabolism.\" Reconstruction of fibrillar cell trajectories showed that, under low temperature stress, hepatopancreas cells had two distinct fates, cell fate 1 and cell fate 2. Cell fate 1 was mainly involved in signal transduction and sensory organ development. Cell fate 2 was mainly involved in metabolic processes. This study preliminarily clarifies the molecular mechanisms underlying cold tolerance in L. vannamei, which will be useful for the breeding of shrimp with greater cold tolerance.

A simple bioassay that quantifies feed intake as an estimation of relative attractability of feeds containing different ingredients in the Pacific white shrimp Litopenaeus vannamei is described. Fish meal (FM), fish protein hydrolysate (FPH), squid meal (SqM) and casein (CN) were assessed at the same dietary level for their relative influence on feed intake rates of Litopenaeus vannamei. A bland diet containing 92% whole wheat grain meal, 6% diatomaceous earth and 2% alginate with a known low attractability was used as the standard control or base diet. Ingredients were added to the bland base control diet at a level of 3% as fed. Shrimp were stocked into 80 L glass tanks (n= 20 per tank) in a recirculating aquaculture system. Tanks were randomly assigned to one of five diet treatments (3tanks/treatment). Experiments measuring the attractability of each feed were conducted twice daily at 0900 hours and 1330 hours over a five day period. For each experiment, 40 feed pellets (ca. 1 g) corresponding to the assigned treatment were provided to each tank. To calculate the rate of feed intake, pellets remaining in each tank were counted at six minute intervals for a seventy-two minute period. Differences in rate of feed intake among diets were evaluated using Cox Regression Analysis. This attractability assay required only small amounts of ingredients and incorporated ingredients into a bland feed, which significantly reduces the influence from other ingredients or compound in the pellets. All of the test protein ingredients, especially SqM, in the feeds significantly increased the feed consumption rate. The diet containing SqM was consumed at a significantly higher rate than those containing casein and FM but not FPH. FPH and CN containing diets were not significantly different but consumed at a higher rate than the diet containing FM. Results of these trials indicate that the presence of certain ingredients can increase feed intake, thereby increasing nutrient availability of the diets. This reported method to determine consumption of diets containing certain ingredients can be considered as a valid method to estimate attractability.

This study aimed to investigate melanosis, quality attributes, and bacterial growth of freeze-chilled Pacific white shrimp (Litopenaeus vannamei) during 6 days of chilled storage, as well as the preservative effects of tea polyphenol on shrimp. The results showed that freeze-chilled storage retarded the growth of bacteria and the accumulation of putrescine in shrimp. The growth of spoilage bacteria Photobacterium and Shewanella were inhibited. However, freeze-chilled storage aggravated melanosis and lipid oxidation. The total volatile basic nitrogen (TVB-N) slightly accumulated in the thawed shrimp. The incorporation of tea polyphenol preserved freeze-chilled shrimp. Melanosis and lipid oxidation of shrimp were alleviated. The accumulation of biogenic amines, TVB-N, hypoxanthine riboside, and hypoxanthine were retarded. Meanwhile, the growth of spoilage bacteria Pseudoalteromonas, Photobacterium, Psychrobacter, and Carnobacterium were inhibited. Based on sensory analysis, the shelf-life of chilled, freeze-chilled, and freeze-chilled tea polyphenol shrimp were 4 days, 3 days, and 6 days, respectively.

This study was performed as part of developing a functional feed ingredient for juvenile Pacific white shrimp (Litopenaeus vannamei). Here we assess the effects of dietary inclusion of a Black Soldier Fly Ingredient (BSFI) from defatted black soldier fly (Hermetia illucens) larvae meal on growth performance, tolerance to salinity stress, and disease resistance when challenged with Vibrio parahaemolyticus or a strain of white spot syndrome virus (WSSV). A control diet was used for comparison with three test diets including 4.5, 7.5, and 10.5% of BSFI (BSFI4.5, BSFI7.5, and BSFI10.5). After 28 days, all diets with BSFI had improved weight gain, feed conversion ratio (FCR) and specific growth rate (SGR) compared to control. Indeed, SGR was significantly improved from inclusion of 4.5% in the diet, whilst FCR was significantly improved at 7.5% (p < 0.05). During the growth trial, survival was not affected by diet. Shrimp health performance was not significantly affected by the diets across the disease and salinity challenges. Overall, the results indicate that the inclusion of BSFI from H. illucens improves the performance of juvenile L. vannamei.

The present study aims to investigate the effects of dietary synbiotics supplementation methods on growth, feed utilization, hepatopancreas and intestinal histology, non-specific immunity and microbiota community of Pacific white shrimp (Litopenaeus vannamei). A control diet was designed to contain 18% fish meal (CON), and then 3 g kg-1 synbiotics (Bioture, consisting of Bacillus subtilis, Saccharomyces cerevisiae, β-glucan and mannan oligosaccharide, etc) was supplemented to the control diet with three methods, directly adding in diets for pelleting (DAP), spraying diets after pelleting at once (SDA), spraying diets before feeding every day (SDE). Shrimp with initial body weight of 1.5 ± 0.12 g were fed one of the four diets for 56 days. The results showed that dietary synbiotics significantly increased the weight gain (WG), apparent digestibility coefficient (ADC) of crude protein (CP) and dry matter (DM), hepatopancreatic protease activity and decreased feed conversion ratio (FCR) (P < 0.05). Among the three synbiotics-added diets, SDE group showed the best growth with significantly higher WG than DAP group (P < 0.05). Serum activities of total superoxide dismutase, catalase, acid phosphatase, lysozyme and alkaline phosphatase of synbiotics-added groups were significantly higher, and serum malondialdehyde level was significantly lower than those of the control (P < 0.05). The intestinal villus width and villus number were also increased by the supplementation of synbiotics. The cumulative mortality was reduced in the three synbiotics-added groups after challenging with Vibrio parahaemolyticus (P < 0.05), and SDE group showed a significantly lower mortality than the control and DAP groups (P < 0.05). In intestinal microbiota composition, the abundance of Lactococcus tended to increase and Vibro tended to decreased in SDA and SDE groups. In conclusion, dietary synbiotics improved the growth, feed utilization, intestine health and non-specific immunity of Pacific white shrimp, and spraying synbiotics on diet presented better performance than adding synbiotics in diet for pelleting.