Stochastic Assembly Increases the Complexity and Stability of Shrimp Gut Microbiota During Aquaculture Progression.

The gut microbiota of aquaculture species contributes to their food metabolism and regulates their health, which has been shown to vary during aquaculture progression of their hosts. However, limited research has examined the outcomes and mechanisms of these changes in the gut microbiota of hosts. Here, Kuruma shrimps from the beginning, middle, and late stages of aquaculture progression (about a time duration of 2 months between each stage) were collected and variations in the gut microbiota of Kuruma shrimp during the whole aquaculture process were examined. High-throughput sequencing demonstrated increases in the diversity and richness of the shrimp gut microbiota with aquaculture progression. In addition, the gut microbiota composition differed among cultural stages, with enrichment of Firmicutes, RF39, and Megamonas and a reduction in Proteobacteria in the mid-stage. Notably, only very few taxa were persistent in the shrimp gut microbiota during the whole aquaculture progression, while the number of taxa that specific to the end of aquaculture was high. Network analysis revealed increasing complexity of the shrimp gut microbiota during aquaculture progression. Moreover, the shrimp gut microbiota became significantly more stable towards the end of aquaculture. According to the results of neutral community model, contribution of stochastic processes for shaping the shrimp gut microbiota was elevated along the aquaculture progression. This study showed substantial variations in shrimp gut microbiota during aquaculture progression and explored the underlying mechanisms regulating these changes.

Effects of chronic dietary hexavalent chromium on bioaccumulation and immune responses in the sea cucumber Apostichopus japonicus.

Sea cucumbers Apostichopus japonicus (3.54 ± 0.01 g of wet weight) were exposed to five concentrations of dietary hexavalent chromium (Cr6+) [0 (control), 100, 200, 400, and 800 mg Cr6+/kg dry weight] amended with K2Cr2O7 for 30 days. The bioaccumulation and immune responses [antioxidant enzymes: superoxide dismutase (SOD) and catalase (CAT); hydrolytic enzymes: acid phosphatase (ACP) and alkaline phosphatase (AKP)] of sea cucumbers were subsequently evaluated. This study found that the order of Cr accumulation in the experimental tissues was respiratory tree > intestine > body wall. Significantly lower SOD activities occurred in the 400 mg/kg group compared to that in the control group. Higher dietary Cr6+ exposure (400 and 800 mg Cr6+/kg dry weight) did not negatively alter the CAT activities, but significantly inhibited CAT activities in 100 mg/kg group, compared to control group. ACP activities in groups 200, 400 and 800 mg/kg were significantly lower than those in control group, while no significant differences occurred in AKP activities among groups. The present study provides important information into the bioaccumulation and immune responses of the sea cucumber A. japonicus in response to chronic dietary Cr6+ exposure.

Effects of chronic prometryn exposure on antioxidative status, intestinal morphology, and microbiota in sea cucumber (Apostichopus japonicus).

Prometryn is an occasional triazine herbicide used in aquaculture to kill algae. However, deposition of prometryn at the bottom of the pond poses a potential threat to aquatic animals, especially benthos, such as the sea cucumber. This study investigated the toxic effects of prometryn oral exposure on antioxidants, and the intestinal histomorphology and microbiome of sea cucumbers. Results showed that the accumulation of prometryn in the intestine, respiratory tree, and body wall decreased sequentially under the same level. Severe pathological damages were observed in the intestines of sea cucumbers fed with 0.080 and 1.595 g/kg prometryn (measured concentration). Moreover, hydrogen peroxide (H2O2) and malondialdehyde (MDA) concentrations were significantly increased in prometryn treatment groups compared to the control group (P < 0.05), while the catalase (CAT) activity was significantly decreased (P < 0.05) in the coelomic fluid of treatment groups. At the phylum level, the abundance of Proteobacteria was significantly higher in the 0.080 g/kg treatment group than in the control group. In addition, prometryn exposure reduced the diversity of intestinal microflora in sea cucumbers. In conclusion, these results suggest that prometryn has potential toxicity to sea cucumber. Therefore, the harm of prometryn deposited in the sediment to aquatic animals must be a concern in aquaculture.

Synergistic Effects of Dietary Selenomethionine and Vitamin C on the Immunity, Antioxidant Status, and Intestinal Microbiota in Sea Cucumber (Apostichopus japonicus).

A 30-day feeding trial was carried out to investigate the interactive effects of dietary selenium (selenomethionine) and vitamin C (Vc) in Apostichopus japonicus. Two selenium (0 and 5 mg/kg) and three vitamin C (0, 5000, and 10,000 mg/kg) combined groups of feed were formulated (Designated as LSeLVc, LSeMVc, LSeHVc, HSeLVc, HSeMVc and HSeHVc, respectively) and fed the sea cucumbers. Our results showed no significant effects on the growth-related parameters in sea cucumber (P > 0.05). Furthermore, the reciprocal action between Se and Vc had significant (P < 0.05) effects on Se accumulation in the respiratory tree and intestines. Also, the lysozyme, glutathione peroxidase activity, and the relative expression levels such as LZM, GPX, Hsp70, and Hsp90 in different tissues were significantly increased in the group of sea cucumber fed diet with 5 mg Se in combination with 5000 mg Vc compared with the control group (P < 0.05). However, MDA and H2O2 contents in the body wall were significantly reduced in the HSeHVc group (P < 0.05). In addition, analysis of intestinal flora revealed that Haloferula abundance was highest in the LSeMVc group than other treatment groups, and Vibrio abundance was decreased with combined Se and Vc supplement. Finally, the species diversity of the gut microbial community of sea cucumber in HSeMVc group was lower than those in other treatment groups. The results showed that the interaction of selenium and vitamin C had positive effects on improving the immune status, antioxidant capacity, and digestive ability of A. japonicus.

The effect of a multi-strain probiotic on growth performance, non-specific immune response, and intestinal health of juvenile turbot, Scophthalmus maximus L.

Probiotic is well known because of its health benefit on the host, including improve growth, treat disease, and enhance immunity. Currently, probiotic has been widely used in aquaculture. However, there is little information about the effect of probiotic on turbot. Therefore, an effort was made to explore the effect of a multi-strain probiotic on growth performance, non-specific immune response, and intestinal health of juvenile turbot, Scophthalmus maximus L. One hundred eighty juvenile turbot (20.04 ± 0.23 g) were randomly divided into three groups (T0, T1, T2), and fed diet were formulated to contain 0%, 1%, and 5% multi-strain probiotic, respectively. Sixty days after the feeding experiment, the growth performance, body composition, enzyme activities, and intestinal microorganism of turbot were analyzed. T2 and T1 showed better growth performance and significant higher (P < 0.05) enzyme activities than T0 (except lysozyme). Moreover, the IV (intestinal villus), IW (intestinal wall), and GC (goblet cell) were well modulated in probiotic treatments. Furthermore, Lactobacillus was found colonized in the intestine of the group fed with 5% multi-strain probiotic. These results suggested adding dietary multi-strain probiotic could positively affect for turbot aquaculture.

Influences of dietary selenomethionine exposure on tissue accumulation, blood biochemical profiles, gene expression and intestinal microbiota of Carassius auratus.

A 30-days feeding trail was conducted to determine the sensitivity of Carassius auratus to the toxicological effects of elevated dietary Selenomethionine (Se-Met). C. auratus averaging 23.56 ±â€¯1.82 g were exposed to four Se-Met concentrations (mg Se/kg): 0 (Se-Met0), 5 (Se-Met5), 10 (Se-Met10) and 20 (Se-Met20) to estimate the effects on tissue selenium (Se) accumulation, blood biochemical profiles, transcript expression and intestinal microbiota. Se accumulated in the kidney, liver and muscle in a dose-dependent manner and followed this order: kidney > liver > muscle, the highest accumulation were obtained in kidney of Se-Met20 diet after 30 days of feeding. Serum contents of alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) in fish exposed to Se-Met20 group was significantly highest among Se-Met exposure groups. Hydrogen peroxide (H2O2) concentrations in liver were affected by dietary Se-Met exposures. Liver contents of total antioxidant capacity (TAC), catalase (CAT), glutathione peroxidase (GPx) and malondialdehyde (MDA) in fish exposure to Se-Met5 group was significantly highest among Se-Met exposure groups. Growth hormone receptor (GHR), insulin-like growth factor 1 (IGF-1) and antioxidant enzyme related genes including glutathione peroxidase (GPx), catalase (CAT) and glutathione S-transferase (GST) expression in liver were down-regulated with the concentration of Se-Met exposure groups. The results of high-throughput sequencing showed that gut microbial communities and hierarchy cluster heatmap analysis were significantly affected by Se-Met exposure. The abundances of Cetobacterium and Vibrio increased while fish exposed to Se-Met20 group. The abundance of Ralstonia increased when the Se-Met exposure dose reached 10 mg Se kg-1. The results suggested that the exposure to elevated dietary Se-Met may result toxic effects in C. auratus.

The Effects of Sub-lethal Dietary Mercury on Growth Performance, Bioaccumulation, and Activities of Antioxidant Enzymes in Sea Cucumber, Apostichopus japonicus.

Triplicate groups of sea cucumbers (4.83 ± 0.15 g) were exposed to one of the four nominal concentrations of dietary mercury [0 (control), 67.6, 338, and 676 mg/kg dry weight, and actually total mercury were 17.55, 87.00, 275.50, 468.50 mg/kg, respectively; Table 3] for 21 days. Mercury accumulation in the intestine showed the greatest mercury burden (77.96 ± 1.20 mg Hg/kg tissue wet weight basis). However, survival rate (SR) was not affected. Body weight gain after the 676 mg Hg/kg treatment was significantly lower than the control group. The feed conversion rate of the 676 mg Hg/kg treatment group was significantly higher than the control group. Additionally, the superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) of sea cucumbers decreased as the mercury dose increased. SOD, T-AOC and alkaline phosphatase of the 676 mg Hg/kg treatment group were significantly lower than the control group. However, there were no significant differences between the four groups in acid phosphatase and catalase (CAT) activity.

Effects of dietary cadmium on growth, antioxidants and bioaccumulation of sea cucumber (Apostichopus japonicus) and influence of dietary vitamin C supplementation.

The effects of dietary cadmium (Cd) supplementation on growth, antioxidant capacity and accumulation of Cd in tissues (body wall, digestive tracts, and respiratory tree) of sea cucumber, Apostichopus japonicus, exposed to sub-chronic concentrations (0, 10, 50, 100, and 500mg Cd/kg dry weight) of Cd were investigated. In addition, the potential protective effects of vitamin C (L-ascorbic acid, AsA) against the effects of Cd on sea cucumbers were investigated. Sea cucumbers were exposed to dietary Cd for 30 days, after which another group of healthy sea cucumbers was supplied diet supplemented with mixed Cd and AsA for another 30 days. Cd exposure for 30 days resulted in increased Cd accumulation in tissues of sea cucumbers with exposure time and concentration. The order of Cd accumulation in organs was digestive tracts>respiratory tree>body wall. On day 30, the body weight gain (BWG) and specific growth rate (SGR) decreased significantly (P<0.05) in the 500mg Cd/kg treatment. Superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) activity and catalase (CAT) activity in the coelomic fluid of sea cucumbers decreased with increasing dietary Cd concentration, but malondialdehyde (MDA) content in the coelomic fluid increased. Providing diet supplemented with Cd and AsA indicated that although sea cucumbers exhibited signs of Cd toxicity, no death occurred in response to 50mg Cd/kg for 30 days. Based on these findings, five treatments were provided: 50mg Cd/kg+0mg AsA/kg, 50mg Cd/kg+ 3000mg AsA/kg, 50mg Cd/kg+ 5000mg AsA/kg, 50mg Cd/kg+10,000mg AsA/kg, and 50mg Cd/kg+15,000mg AsA/kg. The BWG and SGR of sea cucumbers fed the AsA supplemented diet mixed with Cd increased. Additionally, MDA levels in coelomic fluid were negatively correlated with dietary AsA levels, while antioxidant capacities (SOD, GSH-Px and CAT) were positively correlated with dietary AsA levels. Moreover, Cd accumulation in tissues decreased in response to dietary AsA supplementation of treatments. Overall, antioxidant capacity and bioaccumulation in sea cucumber was found to decrease and be induced in response to Cd, but vitamin C mitigated these effects, with 5000mg AsA/kg providing the optimum protection against 50mg/kg Cd.

The effects of dietary lead on growth, bioaccumulation and antioxidant capacity in sea cucumber, Apostichopus japonicus.

Three different diets amended with lead nitrate [Pb(NO3)2] (100, 500 and 1000mg Pb/kg dry weight) and a Pb-free control diet (1.03mg Pb/kg dry weight) were fed to sea cucumber (Apostichopus japonicus) for 30 days. The patterns of Pb accumulation over time were determined in various tissues (body wall, intestine and respiratory tree), as well as growth performance and antioxidant enzymes activities. Pb accumulation in body wall and intestine increased with time in all dietary Pb treatments. When fed the highest Pb diet, the body wall exhibited the greatest Pb burden (16.37mg Pb/kg tissue wet weight), while Pb content in the intestine (2.68mg Pb/kg tissue wet weight) and the respiratory tree (1.78mg Pb/kg tissue wet weight) were lower than Pb content in the body wall by day 30. The body weight gain (BWG), specific growth rate (SGR) and survival rate (SR) had not been affected by 30 days oral administration of Pb supplemented diet. However, the antioxidant enzymes activities [superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px)] of test groups were lower than control group in body wall and malondialdehyde (MDA) concentration in the body wall was opposite after 30 days in sea cucumbers. In summary, this work reports toxic effects in sea cucumber, A. japonicus, after dietary exposure to Pb.

Effects of dietary vitamin C supplementation on lead-treated sea cucumbers, Apostichopus japonicus.

This study aimed to determine the effects of dietary supplementation with vitamin C (l-ascorbic acid, AsA) on lead (Pb) accumulation and toxicity in sea cucumber, Apostichopus japonicus. Three hundred sea cucumbers (10.02±0.02g) fed a basal diet containing 100mg Pb/kg were subjected to 5 levels of l-ascorbate-2-polyphosphate (LAPP) supplementation to achieve 5 AsA treatment levels of 0, 2727, 4630, 9171, 13,893mg AsA/kg. After 30 days, specific growth rate (SGR) and body weight gain (BWG) of supplementation groups were significantly (P<0.05) higher than the control group. Pb contents of the intestine significantly (P<0.05) decreased from 20 days to 30 days in all supplementation groups. Pb contents of the respiratory tree in sea cucumbers in the 2727 and 4630mg AsA/kg treatment groups significantly (P<0.05) decreased from 10 days to 30 days. Superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) activity and total antioxidant capacity (T-AOC) of the sea cucumber body wall in supplementation groups increased after 30 days. Malondialdehyde (MDA) content of the body wall significantly (P<0.05) declined with increasing AsA level. Therefore, it can be demonstrated that vitamin C could mitigate the effects of Pb on sea cucumber and the optimum levels ranged from 2727mg AsA/kg to 4630mg AsA/kg when Pb levels were 100mg/kg.

Alterations of digestive enzyme activities, intestinal morphology and microbiota in juvenile paddlefish, Polyodon spathula, fed dietary probiotics.

The effects of dietary supplementation of probiotics on digestive enzymes activities, intestinal morphology and microbiota in juvenile paddlefish (Polyodon spathula) were studied. A total of 400 fish were reared in two cages and fed with a basal diet (control group, CG) or diet supplemented with commercial probiotics (treatment group, TG) for 80 days. Enzymes activities analysis indicated that protease and α-amylase activities increased (P < 0.01 or P < 0.05) in TG. Light microscopy observation demonstrated the decrease of wall thickness and muscularis thickness in foregut (P < 0.01), the increase of those in hindgut (P < 0.05), the increase of folds height in foregut (P < 0.01) and midgut in TG (P < 0.05). DGGE results of PCR-amplified 16S rRNA confirmed that the richness and diversity of intestinal microbial species increased in TG. The similarity between the commercial bacteria product and intestinal microbiota of TG were higher than the microbiota from CG. The quantities of bacterium, Firmicutes, Proteobacteria, Bacteroidetes, Fusobacteria, present an increasing trend from foregut to hindgut both in two groups. To our knowledge, this is the first in vivo study to reveal the effect of dietary probiotics on intestinal digestive enzymes activities, morphology and microbiota in paddlefish.

Effects of palm oil blended with oxidized fish oil on growth performances, hematology, and several immune parameters in juvenile Japanese sea bass, Lateolabrax japonicas.

A 60-day feeding trial was conducted to determine the effects of palm oil blended with oxidized and non-oxidized fish oil on growth performances, hematology, and non-specific immune response in juvenile Japanese sea bass, Lateolabrax japonicas. Japanese sea bass (1.73 ± 0.01 g) were fed seven experimental diets containing 100 g/kg of dietary lipid in forms of palm oil (10P), fish oil (10F), fish oil blended with palm oil at different ratios, 6:4 (6F4P) and 4:6 (4F6P), oxidized fish oil (10OF), and oxidized fish oil blended with palm oil at different ratios, 6:4 (6OF4P) and 4:6 (4OF6P). After the feeding trial, the following results were illustrated. No significant effects were observed in survival, feed conversion ratio, condition factor, and hematocrit after feeding with experimental diets for 60 days. The relatively higher specific growth rate and hematology were observed in 6F4P. Furthermore, both palm oil and oxidized fish oil acted as a negatively on serum lysozyme activity (P < 0.05). This study suggested that a ration of 6F4P is recommended as an innocuous ratio for Japanese sea bass. Furthermore, according to the present investigation, palm oil seems to have the ability to improve the protein efficiency when added to oxidized fish diets as well as a positive trend to the growth performance (P > 0.05).

Enhancement of non-specific immune response in sea cucumber (Apostichopus japonicus) by Astragalus membranaceus and its polysaccharides.

In this study, the immunostimulatory effect of oral administration of different preparations (conventional fine powder [CP] and superfine powder [SP]) of Astragalus membranaceus root or its polysaccharides (APS) in sea cucumber (Apostichopus japonicus) was investigated. Sea cucumbers with an average initial weight of 49.3 +/- 5.65 g were fed with a diet containing 3% CP or SP or 0.3% APS over a period of 60 days. The non-specific humoral (phenoloxidase, lysozyme and agglutination titer) and cellular (phagocytic capacity and reactive oxygen species) responses were determined and compared with controls (no supplement) after 20, 40 and 60 days of feeding. Variation in the levels of responses was evident among different supplements. SP and APS significantly enhanced most of the immune parameters tested. Among the humoral responses, lysozyme activity significantly increased after feeding with SP-supplemented diet for 20, 40 or 60 days. Furthermore, lectin titer showed significant enhancement after 20 and 60 days of feeding with APS-supplemented diet. Significant increase in the production of reactive oxygen species was evident for all three supplements after 20 days of feeding, but no significant change in serum phenoloxidase activity was observed for any of the three supplements over the three different periods. Overall, significant modulation of the cellular responses was only noticed after 20 days of feeding with SP- or APS-supplemented diet. After 60 days, these two groups also exhibited a decrease in the cumulative symptom rates compared to the controls when challenged with Vibrio splendidus. These results indicated that dietary intake containing A. membranaceus root or its polysaccharides could enhance the immune responses of A. japonicus and improve its resistance to infection by V. splendidus.