Host Species and Environment Shape the Gut Microbiota of Cohabiting Marine Bivalves.

Pacific oysters (Crassostrea gigas) and Mediterranean mussels (Mytilus galloprovincialis) are commercially important marine bivalves that frequently coexist and have overlapping feeding ecologies. Like other invertebrates, their gut microbiota is thought to play an important role in supporting their health and nutrition. Yet, little is known regarding the role of the host and environment in driving these communities. Here, bacterial assemblages were surveyed from seawater and gut aspirates of farmed C. gigas and co-occurring wild M. galloprovincialis in summer and winter using Illumina 16S rRNA gene sequencing. Unlike seawater, which was dominated by Pseudomonadata, bivalve samples largely consisted of Mycoplasmatota (Mollicutes) and accounted for >50% of the total OTU abundance. Despite large numbers of common (core) bacterial taxa, bivalve-specific species (OTUs) were also evident and predominantly associated with Mycoplasmataceae (notably Mycoplasma). An increase in diversity (though with varied taxonomic evenness) was observed in winter for both bivalves and was associated with changes in the abundance of core and bivalve-specific taxa, including several representing host-associated and environmental (free-living or particle-diet associated) organisms. Our findings highlight the contribution of the environment and the host in defining the composition of the gut microbiota in cohabiting, intergeneric bivalve populations.

Dietary selenium supplementation alleviates low salinity stress in the Pacific white shrimp Litopenaeus vannamei: growth, antioxidative capacity and hepatopancreas transcriptomic responses.

Se is an essential trace element associated with animal growth and antioxidant and metabolic processes. However, whether Se, especially organic Se with higher bioavailability, can alleviate the adverse effects of low salinity stress on marine economic crustacean species has not been investigated. Accordingly, juvenile Pacific white shrimp (Litopenaeus vannamei) were reared in two culture conditions (low and standard salinity) fed diets supplemented with increasing levels of l-selenomethionine (0·41, 0·84 and 1·14 mg/kg Se) for 56 d, resulting in four treatments: 0·41 mg/kg under standard seawater (salinity 31) and 0·41, 0·84 and 1·14 mg/kg Se under low salinity (salinity 3). The diet containing 0·84 mg/kg Se significantly improved the survival and weight gain of shrimp under low salinity stress and enhanced the antioxidant capacity of the hepatopancreas. The increased numbers of B and R cells may be a passive change in hepatopancreas histology in the 1·14 mg/kg Se group. Transcriptomic analysis found that l-selenomethionine was involved in the regulatory pathways of energy metabolism, retinol metabolism and steroid hormones. In conclusion, dietary supplementation with 0·84 mg/kg Se (twice the recommended level) effectively alleviated the effects of low salinity stress on L. vannamei by regulating antioxidant capacity, hormone regulation and energy metabolism.

Growth, Health, and Gut Microbiota of Female Pacific White Shrimp, Litopenaeus vannamei Broodstock Fed Different Phospholipid Sources.

Phospholipids have an important antioxidant effect on animals. The effects of different dietary phospholipid sources on the growth, antioxidant activity, immunity, and gut microbiota of female broodstock of Pacific white shrimp Litopenaeus vannamei were investigated. Four isoproteic and isolipid semi-purified diets containing 4% soybean lecithin (SL), egg yolk lecithin (EL), or krill oil (KO) and a control diet without phospholipid supplementation were fed to female broodstock of L. vannamei (34.7 ± 4.2 g) for 28 days. The growth performance, antioxidative capacity, and innate immunity of the female broodstock fed phospholipid supplemented diets were improved regardless of sources compared with the control shrimp. The effects on growth and antioxidant capacity in female shrimp fed the KO diet were highest. The innate immunity of female shrimp fed the EL and KO diets were significantly higher than shrimp fed the SL diet. Dietary phospholipid supplementation increased gut microbiota diversity and richness, and the Chao1 and ACE values in the KO group were significantly higher than in the control group. The richness of Proteobacteria, Photobacterium, and Vibrio decreased, whereas the richness of Firmicutes and Bacteroidetes increased in the shrimp fed the KO diet compared with the shrimp fed the SL and EL diets. The interactions of gut microbiota in shrimp fed the KO diet were the most complex, and the positive interaction was the largest among all the treatments. The functional genes of gut microbiota in shrimp fed the KO diet were significantly enriched in lipid metabolism and terpenoid/polyketide metabolism pathways. Spearman correlation analysis showed that Fusibacter had significantly positive correlations with antioxidant activity (total antioxidant capacity, superoxide dismutase, glutathione peroxidase), immune enzyme activity (phenoloxidase and lysozyme), and immune gene expression (C-type lectin 3, Caspase-1). All findings suggest that dietary phospholipids supplementation can improve the growth and health status of female L. vananmei broodstock. Krill oil is more beneficial in improving the antioxidant capacity and innate immunity than other dietary phospholipid sources. Furthermore, krill oil can help establish the intestinal immune barrier by increasing the richness of Fusibacter and promote the growth of female shrimp. Fusibacter may be involved in iron metabolism to improve the antioxidant capacity of female shrimp.

Combined toxic effects of thiamethoxam on intestinal flora, transcriptome and physiology of Pacific white shrimp Litopenaeus vannamei.

The environmental accumulation of thiamethoxam has increasingly become a risk for the health of aquatic animals, especially crustacean species in the same phylum as the target pests. The lack of knowledge on the toxicity of thiamethoxam to crustaceans motivates our research to study the acute and chronic toxicity of decapod crustaceans Litopenaeus vannamei, exposed to thiamethoxam. A 28-day chronic toxicity test followed a 96 h acute toxicity test. Thiamethoxam induced oxidative stress and decreased growth performance in shrimp. In addition, thiamethoxam has led to a substantial imbalance of the micro-ecosystem in the intestine. The composition of the intestinal flora changed significantly, and the balance of the interaction network in genera was broken. The competitive interaction of many bacteria becomes an unstable cooperative interaction. Transcriptomic analysis showed that the numbers of up- and down-regulated differentially expressed genes (DEGs) increased in a dose-dependent manner. These DEGs were significantly enriched in pathways related to detoxification, and the expression of most detoxification genes was upregulated. DEGs related to detoxification were positively correlated with Shimia and negatively correlated with Pseudoalteromonas. This study provides evidence for the first time on the toxic effects of thiamethoxam on the growth, biochemistry, intestinal flora, and transcriptome in crustaceans.

Effects and Mechanism of Different Phospholipid Diets on Ovary Development in Female Broodstock Pacific White Shrimp, Litopenaeus vannamei.

Research on nutrition and feed development for the broodstock of the Pacific white shrimp, Litopenaeus vannamei, is rare, and a poor broodstock quality is a critical factor restricting the seed supply in shrimp farming. As an essential nutrient for the gonadal development of L. vannamei, one control diet (no phospholipid) and three typical phospholipids (soybean lecithin, egg yolk lecithin, and krill oil) were evaluated in a semipurified diet of 4% phospholipid for a 28-day trial (initial weight 34.7 ± 4.2 g). Dietary phospholipid supplementation significantly promoted the ovarian maturation of female L. vannamei. Compared with soybean lecithin and egg yolk lecithin, krill oil showed the best positive results. Shrimp fed with a diet krill oil has obtained a significantly higher gonadosomatic index, yolk particle deposition, lipid accumulation, and estrogen secretion than from other sources. Ovary lipidomic analysis showed that the krill oil enriched the lipid composition of the ovary. The "glycerophospholipid metabolism" and "sphingolipid metabolism" pathways were significantly varied via topological pathway analysis. Genes and hub genes, with significantly different expression levels, were significantly enriched in the "fatty acid metabolism pathway," "glycerophospholipid metabolism," and "arachidonic acid metabolism" pathways by transcriptomic analysis. Correlation analysis of the transcriptome and lipidomics showed that the differential gene "hormone-sensitive lipase-like" (HSL) was positively correlated with various lipids [triglycerides (TG), phosphatidic acid (PA), phosphatidylserine (P), phosphatidylethanolamine (PE), glucosylceramide (GlcCer), phosphatidylglycerol (PG), and phosphatidylinositol (PI)] but was negatively correlated with diacylglycerol (DG), lysophosphatidylethanolamine (LPE), and sphingomyelin (SM). In conclusion, the dietary phospholipids, especially krill oil as a phospholipid source, can promote the development of L. vannamei ovaries by increasing the accumulation of nutrients such as triglycerides and sterols, and the secretion of estrogen or related hormones, such as estradiol and methylfarneside, by affecting the metabolism of glycerol phospholipids and some key fatty acids.

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 imidacloprid exposure on the biochemical responses, transcriptome, gut microbiota and growth performance of the Pacific white shrimp Litopenaeus vannamei.

The widespread use of neonicotinoid insecticides, such as imidacloprid, in agriculture is one of the key factors for the drop in the survival of invertebrates, including decapod crustaceans. However, there is currently a lack of comprehensive studies on the chronic toxicity mechanisms in decapod crustaceans. Here, the concentration-dependent effects of imidacloprid on the physiology and biochemistry, gut microbiota and transcriptome of L. vannamei , and the interaction between imidacloprid, gut microbiota and genes were studied. Imidacloprid caused oxidative stress, leading to reduced growth and to immunity and tissue damage in L. vannamei . Imidacloprid increased the gut pathogenic microbiota abundance and broke the steady state of the gut microbiota interaction network, resulting in microbiota function disorders. Chronic imidacloprid exposure induced overall transcriptome changes in L. vannamei . Specifically, imidacloprid caused a large number of differentially expressed genes (DEGs) to be significantly downregulated. The inhibition of autophagy-related pathways revealed the toxic process of imidacloprid to L. vannamei . The changes in phase I and II detoxification gene expression clarified the formation of a detoxification mechanism in L. vannamei . The disturbance of circadian rhythm (CLOCK) caused by imidacloprid is one of the reasons for the increase in gut pathogenic microbiota abundance.

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.

Acute acidification stress weakens the head kidney immune function of juvenile Lates calcarifer.

Acidized water environment can impact many physiological processes of aquatic animals. The response of the head kidney to acidification, especially the immune response, is of great significance to health. This study analyzed the histological and transcriptional changes under different acidification levels (C group, pH 8.1; P group, pH 7.4; E group, pH 3.5) in the short term (12 h, 36 h and 60 h) in the head kidney of juvenile L. calcarifer. The results showed that the acidification of the water environment caused tissue damage to the head kidney of L. calcarifer, and the damage appeared earlier and was stronger in the extreme pH group. The transcriptional response of L. calcarifer head kidney increased with the increase of acidification level. The two treatments transcriptional responses showed different trends in terms of time. After KEGG function enrichment, with the increase of stimulation time, the proportion of down-regulated pathways was increasing, and the types of pathway enrichment at different acidification levels were quite different at the initial stage. At 12 h, the first category in the P group with the most significant number of pathways was 'Metabolism', and the first category in the E group with the largest number of pathways was 'Human Diseases'. At 60 h, the enrichment pathways of the two groups were highly overlapping in immune-related pathways, which contained 26 common DEGs. They had a dominant expression pattern. In the P group, the expression level decreased with time. In the E group, the down-regulation degree of expression level at 12 h reached the level of the P group at 60 h, and the expression level remained low until 60 h. Through the correlation network, interferon regulatory factor 7 (IRF7), Tripartite motif containing-21 (TRIM21), Signal transducer and activator of transcription 1 (STAT1) and Signal transducer and activator of transcription 3 (STAT3) were found to have the most correlation with other genes. In this study, juvenile L. calcarifer showed different coping strategies to different levels of acute acidification stress, but all of them resulted in the extensive weakening of head kidney immune function.

Influences of dietary vitamin D3 on growth, antioxidant capacity, immunity and molting of Chinese mitten crab (Eriocheir sinensis) larvae.

This study investigates the effects of vitamin D3 (VD3) on growth performance, antioxidant capacity, immunity and molting of larval Chinese mitten crab Eriocheir sinensis. A total of 6,000 larvae (7.52 ± 0.10 mg) were fed with six isonitrogenous and isolipidic experimental diets with different levels of dietary VD3 (0, 3000, 6000, 9000, 12000 and 36000 IU/kg) respectively for 23 days. The highest survival and molting frequency were found in crabs fed 6000 IU/kg VD3. Weight gain, specific growth rate, and carapace growth significantly increased in crabs fed 3000 and 6000 IU/kg VD3 compared to the control. Broken-line analysis of molting frequency, weight gain and specific growth rate against dietary VD3 levels indicates that the optimal VD3 requirement for larval crabs is 4825-5918 IU/kg. The highest whole-body VD3 content occurred in the 12000 IU/kg VD3 group, and the 25-dihydroxy VD3 content decreased with the increase of dietary VD3. The malonaldehyde content was lower than the control. Moreover, the superoxide dismutase activity, glutathione peroxidase and total antioxidant capacity of crab fed 6000 IU/kg VD3 were significantly higher than in control. Crabs fed 9000 IU/kg showed the highest survival after 120 h of salinity stress, and the relative mRNA expressions indicate vitamin D receptor (VDR) is the important regulatory element in molting and innate immunity. The molting-related gene expressions showed that the response of crab to salinity was self-protective. This study would contribute to a new understanding of the molecular basis underlying molting and innate immunity regulation by vitamin D3 in E. sinensis.

Recovery from Hypersaline-Stress-Induced Immunity Damage and Intestinal-Microbiota Changes through Dietary ß-glucan Supplementation in Nile tilapia (Oreochromis niloticus).

Long-term exposure to hyperosmotic environments can induce severe immune damage and increase risk in tilapia breeding. As an effective immunoregulator, ß-glucan has attracted extensive attention in nutritional research and given rise to high expectations of improving health status and alleviating organismal damage in tilapia, Oreochromis niloticus, in brackish water. In this study, an 8-week cultivation experiment was conducted on tilapia fed a basal diet or diets with ß-glucan supplementation in freshwater (control) and brackish water. Growth performance, hematological aspects, immune cytokine expression, and the intestinal microbiota of tilapia were analyzed. The results indicated that supplementation with ß-glucan significantly reduced the enlarged spleen of tilapia resulting from hypersaline stress. Tilapia fed ß-glucan showed significantly-greater decreases in the red blood cell count, hematocrit, red cell distribution width, platelet count, and plateletcrit than those fed the basal diet. ß-glucan significantly decreased the high expression of immune-related genes in the spleen induced by hyperosmotic stress. In the intestine, the high migration inhibitory factor-2 (MIF-2) and IL-1ß gene expression induced by hypersaline stress was significantly reduced. ß-glucan supplementation also significantly increased the abundance of beneficial microbiota such as Lactobacillus, Phycicoccus, and Rikenellaceae. Therefore, dietary ß-glucan supplementation can significantly reduce spleen enlargement and improve immune function in tilapia in brackish water. ß-glucan intake can also optimize the intestinal microbiota of tilapia in brackish water and improve fish health.

Growth, Metabolite, Antioxidative Capacity, Transcriptome, and the Metabolome Response to Dietary Choline Chloride in Pacific White Shrimp Litopenaeus vannamei.

To determine the response of Pacific white shrimp Litopenaeus vannamei to different levels of dietary choline, juvenile white shrimp (1.75 ± 0.09 g) were fed six semi-purified diets supplemented with 0 (control), 2000, 4000, 6000, 8000, and 12,000 mg/kg choline chloride for eight weeks. Growth performance, whole-body composition, serum characteristics and hepatopancreatic antioxidant indexes were evaluated. Meanwhile, serum metabolome and hepatopancreas transcriptome were performed to examine the overall difference in metabolite and gene expression. The weight gain, survival, specific growth rate, condition factor and hepatosomatic index were not affected by dietary choline levels. The shrimp fed 6000 mg/kg dietary choline chloride gained the maximal whole-body crude protein, which was significantly higher than that of shrimp fed with 12,000 mg/kg dietary choline. Serum total cholesterol of shrimp fed 6000 mg/kg dietary choline was higher than that in shrimp fed 4000 mg/kg choline. Dietary choline significantly decreased malondialdehyde content, superoxide dismutase, and glutathione peroxidase activities in shrimp hepatopancreas. Compared with the shrimp fed 6000 mg/kg dietary choline chloride, the glycerophospholipid metabolism pathway was significantly enriched in the shrimp fed 0 mg/kg dietary choline chloride, and the choline content and bile salt-activated lipase-like expression were upregulated. The expression of trypsin-1-like in protein digestion and absorption pathway was significantly downregulated in the shrimp fed 12,000 mg/kg dietary choline chloride. Apolipoprotein D might be a potential biomarker in shrimp, and dietary choline played an important role in lipid metabolism, especially in the reduction of oxidative damage in L. vannamei. Based on the results of weight gain and degree of oxidative damage, 1082 mg/kg dietary choline could meet the growth requirement of L. vannamei, but 2822 mg/kg dietary choline was needed to reduce peroxidation damage.

A Comparative Study on Growth and Metabolism of Eriocheir sinensis Juveniles Under Chronically Low and High pH Stress.

This study elucidates the effects of chronic pH stress on the growth and metabolic response of juvenile Chinese mitten crab Eriocheir sinensis. Crabs were exposed under normal pH (control, pH = 8.0 ± 0.20), low pH (pH = 6.5 ± 0.20), and high pH (pH = 9.5 ± 0.20) in an 8-week trial. Both low and high pH suppressed weight gain but low pH had more adverse effects. No difference was observed on survival, crude lipid, and protein. Acidic stress significantly reduced protein efficiency. The malondialdehyde (MDA) content in hepatopancreas was highest at low pH. The superoxide dismutase (SOD) activity in hepatopancreas and total hemocyte counts (THC) in the stress groups were higher than that in the control. Crabs under high pH had the highest ACP and AKP activities, but there was no significant difference between the control and low pH groups. In the transcriptome analysis, 500.0M clean reads were obtained from the control, low pH, and high pH groups, and assembled into 83,025 transcripts. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were analyzed to obtain the significantly changed pathways involving differently expressed genes. Ten and eight pathways in metabolism were significantly changed in low pH vs control and high pH vs control groups, respectively. According to the reported functions of these pathways, most of them participated in carbohydrate metabolism. The metabolism pathway analysis indicates the increases of stress resistance, glucose metabolism, and molting activities under chronically pH stress. This study suggests that low pH has more negative impact on crab growth, and oxidative phosphorylation is the main source of energy source under low pH stress, while aerobic glycolysis supplies most energy under high pH stress.

Effects of dietary T-2 toxin on gut health and gut microbiota composition of the juvenile Chinese mitten crab (Eriocheir sinensis).

The current study aims to investigate the effects of dietary T-2 toxin on the intestinal health and microflora in the juvenile Chinese mitten crab (Eriocheir sinensis) with an initial weight 2.00 ± 0.05 g. Juvenile crabs were fed with experimental diets supplemented with T-2 toxin at 0 (control), 0.6 (T1 group), 2.5 (T2 group) and 5.0 (T3 group) mg/kg diet for 8 weeks. Dietary T-2 toxin increased the malondialdehyde (MDA) content and the expression of Kelch-like ECH-associated protein 1 (keap1) gene while the expression of cap 'n' collar isoform C (CncC) decreased in the intestine. The activities of glutathione peroxidase (GSH-Px) and total anti-oxidation capacity (T-AOC) in the intestine increased only in the lower dose of dietary T-2. Dietary T-2 toxin significantly increased the mRNA expression of caspase-3, caspase-8, Bax and mitogen-activated protein kinase (MAPK) genes and the ratio of Bax to Bcl-2 accompanied with a reduction of Bcl-2 expression. Furthermore, T-2 toxin decreased the mRNA levels of antimicrobial peptides (AMPs), peritrophic membrane (PM1 and PM2) and immune regulated nuclear transcription factors (Toll-like receptor: TLR, myeloid differentiation primary response gene 88: Myd88, relish and lipopolysaccharide-induced TNF-α factor: LITAF). The richness and diversity of the gut microbiota were also affected by dietary T-2 toxin in T3 group. The similar dominant phyla in the intestine of the Chinese mitten crab in the control and T3 groups were found including Bacteroidetes, Firmicutes, Tenericutes and Proteobacteria. Moreover, the inclusion of dietary T-2 toxin of 4.6 mg/kg significantly decreased the richness of Bacteroidetes and increased the richness of Firmicutes, Tenericutes and Proteobacteria in the intestine. At the genus level, Dysgonomonas and Romboutsia were more abundant in T3 group than those in the control. However, the abundances of Candidatus Bacilloplasma, Chryseobacterium and Streptococcus in T3 group were lower than those in the control. This study indicates that T-2 toxin could cause oxidative damage and immunosuppression, increase apoptosis and disturb composition of microbiota in the intestine of Chinese mitten crab.

Gemfibrozil improves lipid metabolism in Nile tilapia Oreochromis niloticus fed a high-carbohydrate diet through peroxisome proliferator activated receptor-α activation.

High carbohydrate diet (HCD) can induce lipid metabolism disorder, characterized by excessive lipid in farmed fish. Peroxisome proliferator activated receptor-α (PPARα) plays an important role in lipid homeostasis. In this study, we hypothesize that PPARα can improve lipid metabolism in fish fed HCD. Fish (3.03 ± 0.11 g) were fed with three diets: control (30% carbohydrate), HCD (45% carbohydrate) and HCG (HCD supplemented with 200 mg/kg gemfibrozil, an agonist of PPARα) for eight weeks. The fish fed HCG had higher growth rate and protein effiency than those fed the HCD diet, whereas the opposite trend was observed in feed conversion ratio, hepatosomatic index and mesenteric fat index. Additionally, fish fed HCG significantly decreased lipid accumulation in the whole body, liver and adipose tissues compared to those fed the HCD diet. Furthermore, fish in the HCG group significantly increased the mRNA and protein expression and protein dephosphorylation of PPARα. The HCG group also significantly increased the mRNA level of the downstream target genes of PPARα, whereas the opposite trend occured in the mRNA level of lipolysis-related genes compared to the HCD group. Besides, fish in the HCG group remarkably decreased the contents of alanine aminotransferase, aspartate aminotransferase and malondialdehyde, whereas the opposite occured in the activities of antioxidative enzymes and anti-inflammatory cytokine genes compared to the HCD group. This study indicates that gemfibrozil can improve lipid metabolism and maintain high antioxidant and anti-inflammatory capacity through activating PPARα in Nile tilapia fed a high carbohydrate diet.

Transcriptional analysis reveals physiological response to acute acidification stress of barramundi Lates calcarifer (Bloch) in coastal areas.

To understand the physiological response of estuarine fish to acidification, barramundi (Lates calcarifer) juveniles were exposed to acidified seawater in experimental conditions. The molecular response of barramundi to acidification stress was assessed by RNA-seq analysis. A total of 2188 genes were identified as differential expression genes. The gene ontology classification system and Kyoto Encyclopedia of Genes and Genomes database analysis showed that acidification caused differential expressions of genes and pathways in the gills of barramundi. Acidification had a great influence on the signal transduction pathway in cell process. Furthermore, we detected that numerous unigenes involved in the pathways associated with lipid metabolism, carbohydrate metabolism, amino acid metabolism, glycan biosynthesis and metabolism specific and non-specific immunity were changed. This study indicates that the physiological responses in barramundi especially the immune system and energy allocation correspond to the variation of environmental pH. This study reveals the necessity for assessment of the potential of estuarine fishes to cope with acidification of the environment and the need to develop strategies for fish conservation in coastal areas.

Sodium butyrate can improve intestinal integrity and immunity in juvenile Chinese mitten crab (Eriocheir sinensis) fed glycinin.

Butyrate is a fermentation byproduct of gut microbiota and is susceptible to chronic oxidative stress. This study investigates the mitigative effects of sodium butyrate (SBT) on growth inhibition and intestinal damage induced by glycinin in juvenile Chinese mitten crab (Eriocheir sinensis). All four experimental diets containing 80 g/kg glycinin were formulated with 0, 10, 20 and 40 g/kg SBT respectively. There was no glycinin or SBT in the control diet. Juvenile crabs (0.33 ± 0.01g) were respectively fed with these five diets for eight weeks. The diets with 10 and 20 g/kg SBT significantly improved the survival and weight gain of the crabs compared with those in the 0 g/kg SBT group, and showed no difference with the control group. The crabs fed diets containing glycinin without SBT had lower glutathione and glutathione peroxidase activities but higher malondialdehyde in the intestine than those in the control group. Moreover, dietary glycinin decreased the lysozyme and phenoloxidase activities and improved the level of histamine in the intestine compared with the control group, while the supplementation of SBT counteracted these negative effects. The addition of SBT could also restore the impaired immunity and morphological structure of the intestine. Dietary SBT could increase the mRNA expression of antimicrobial peptides genes (anti-lipopolysaccharide factor 1 and 2) and decrease the content of pro-inflammatory factor TNF-α. The SBT could restore the intestinal microbial community disorganized by glycinin. The abundance of pathogenic bacteria (Aeromonas, Vibrio and Pseudomonas) decreased significantly and the potential probiotic bacteria (Bacillus, Lactobacillus, Chitinibacter and Dysgonomonas) increased significantly in the 10 g/kg SBT group. This study suggests that sodium butyrate supplementation can mitigate the negative effects induced by glycinin such as growth inhibition, intestinal inflammation and reduction of beneficial flora in the gut.

Could the gut microbiota community in the coral trout Plectropomus leopardus (Lacepède, 1802) be affected by antibiotic bath administration?

Gut microbiota in fish plays an important role in the nutrient digestion, immune responses and disease resistance. To understand the effect of fluoroquinolone antibiotic bath administration on fish gut microbiota, the gut microbiota community in the coral trout Plectropomus leopardus (Lacepède, 1802) was studied after enrofloxacin bathing treatment at two concentrations (5 and 10 mg/L) and 0 mg/L as control. A total of 90 fish were used in this study, and three replicates were used for each treatment. After a 24-hr bath, the gut bacterial composition was analyzed using high-throughput Illumina sequencing. The results indicated that the richness, diversity and the dominant bacterial taxa of P. leopardus gut bacteria were not affected by enrofloxacin bathing (p > .05). Proteobacteria and Firmicutes were the dominant phyla, and Exiguobacterium, Citrobacter, Vibrio, Acinetobacter, Pseudomonas were the dominant genus. The findings in the present study provide an understanding on the relationship between fish gut bacteria community and antibiotic bath administration. The findings of this study are instructive on the antibiotic bath administration applied for the management of P. leopardus health in aquaculture.

Growth and health status of Pacific white shrimp, Litopenaeus vannamei, exposed to chronic water born cobalt.

Cobalt (Co) is an important component of vitamin B12, but is toxic to aquatic animals at a high level. In this study, the Pacific white shrimp, Litopenaeus vannamei were exposed to three Co concentrations (0, 100, and 1000 µg/L) for 4 weeks. The survival and condition factor in shrimp exposed to the Co treatments were not different from the control, but the shrimp exposed to 100 µg Co/L gained more weight than in other two groups, and the shrimp exposed to 1000 µg Co/L gained less weight than in other groups. The SOD and GSH-PX activities were higher in shrimp exposed to 100 µg Co/L, but lower in the shrimp exposed to 100 µg Co/L compared with the control, respectively. The MDA contents in the hepatopancreas decreased in the 100 µg Co/L, but increased in the 1000 µg Co/L. The serum lysozyme decreased with ambient cobalt, was lower in the shrimp exposed to 1000 µg Co/L than in other two groups. The expression of C-type lectin 3 was down-regulated by Co concentrations. The Toll and immune deficiency in shrimp exposed to 100 µg Co/L was higher than in other two groups. The mucin-1 was lower in the 1000 µg Co/L group than in other two groups, but mucin-2 and mucin-5AC were higher in the 1000 µg Co/L group than in the control. With increasing Co concentration, Shannon and Simpson indexes of the intestinal microbial communities were decreased. The abundance of pathogenic bacteria (Ruegeria and Vibrio) increased in both Co groups. This study indicates that chronic exposure to waterborne cobalt could affect growth, cause oxidative stress, stimulate the immune response, damage intestinal histology, and reshape intestinal microbiota community L. vannamei.

Toxic effect of chronic waterborne copper exposure on growth, immunity, anti-oxidative capacity and gut microbiota of Pacific white shrimp Litopenaeus vannamei.

Copper can be accumulated in water through excessive sewage discharge or residual algaecide to generate toxic effect to aquatic animals. In this study, the juvenile of Pacific white shrimp, Litopenaeus vannamei was exposed to 0 (control), 0.05, 0.1, 0.2, 0.5 or 1 mg Cu2+ L-1 for 30 days. Growth, immune function, anti-oxidative status and gut microbiota were evaluated. Weight gain and specific growth rate of L. vannamei were significantly decreased with the increase of ambient Cu2+. Enlarged lumen and ruptured cells were found in the hepatopancreas of shrimp in the 0.5 or 1 mg Cu2+ L-1 treatment. Total hemocyte counts of shrimp in 0.5 or 1 mg Cu2+ L-1 were significantly lower than in the control. The hemocyanin concentration was also significantly increased in 0.2 or 0.5 mg Cu2+ L-1. Lysozyme contents were reduced in shrimp when Cu2+ exceeded 0.2 mg L-1. Meanwhile, activities of superoxide dismutase and glutathione peroxidase were increased in the hepatopancreas and the activity of Na+-K+ ATPase was decreased in the gills with increasing Cu2+. The mRNA expressions of immune deficiency, toll-like receptor and caspase-3 were all significantly higher in the hepatopancreas in 0.05 mg Cu2+ L-1 than in the control. For the diversity of intestinal microbes, Bacteroidetes significantly decreased in 1 mg Cu2+ L-1 at the phylum level. KEGG pathway analysis demonstrates that 1 mg L-1 Cu2+ can significantly alter metabolism, cellular processes and environmental information processing. This study indicates that the concentration of 1 mg L-1 Cu can negatively impact growth, hemolymph immunity, anti-oxidative capacity and gut microbiota composition of L. vannamei.