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.

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.

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.

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.

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.

T-2 toxin in the diet suppresses growth and induces immunotoxicity in juvenile Chinese mitten crab (Eriocheir sinensis).

The T-2 toxin is a trichothecene mycotoxin and is highly toxic to aquatic animals, but little is known on its toxic effect in crustaceans. In the present study, the crab juveniles were fed with diets containing four levels of T-2 toxin: 0 (control), 0.6 (T1), 2.5 (T2) and 5.0 (T3) mg/kg diet for 56 days to evaluate its impact on the juvenile of Chinese mitten crab (Eriocheir sinensis). The crabs fed the T-2 toxin diets had significantly lower weight gain and specific growth rate than those fed the control diet. Moreover, crab survival in T3 group was obviously lower than that in the control. Oxidative stress occurred in all the treatment groups as indicated by higher activities of total superoxide dismutase, glutathione peroxidase, and total antioxidant capacity than those in the control. The total hemocyte count, respiratory burst, phenoloxidase in the hemolymph, and phenoloxidase, acid phosphatase and alkaline phosphatase in the hepatopancreas of crabs fed T-2 toxin were significantly lower than those in the control. The transcriptional expressions of lipopolysaccharide-induced TNF-alpha factor, relish, and the apoptosis genes in the hepatopancreas were induced by dietary T-2 toxin. The genes related to detoxication including cytochrome P450 gene superfamily and glutathione S transferase were induced in low concentration, then decreased in high concentration. Dietary T-2 toxin damaged the hepatopancreas structure, especially as seen in the detached basal membrane of hepatopancreatic tubules. This study indicates that dietary T-2 toxin can reduce growth performance, deteriorate health status and cause hepatopancreas dysfunction in crabs.

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.

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.

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.

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.

Dietary supplementation of selenium yeast enhances the antioxidant capacity and immune response of juvenile Eriocheir Sinensis under nitrite stress.

This study elucidates the response to nitrite stress and the effect of dietary selenium supplements on the growth, antioxidant activity, immunity and transcriptome of juvenile Chinese mitten crab Eriocheir sinensis. In the control group, the crabs were fed the diet without selenium supplementation and there was no nitrite addition to the water. In the test group, the crabs were fed diets with three levels of selenium 0 (N1), 0.5 (N2) and 1.0 (N3) mg/kg in the water containing 2 mg/L NO2N as a stress factor for eight weeks. Feed conversion ratio (FCR) was improved by adding dietary selenium. There was no significant difference in specific growth rate and weight gain between N1 and the control groups, or among different selenium levels in the test group. The superoxide dismutase (SOD) activity was significantly lower, but malondialdehyde (MDA) was higher in the N1 group than those in the serum and hepatopancreas of the control group. The activities of SOD, glutathione peroxidase (GPx) and acid phosphatase increased at the medium level of selenium but decreased as the level of dietary selenium increased to 1.0 mg/kg. The serum lysozyme (LZM) activity increased but the MDA content in both serum and hepatopancreas decreased with the increase of selenium levels. The total clean reads of the crabs in the control group, N1 and N3 groups reached 390.7M and were assembled into 106 471 transcripts. Compared with the control group, 1196 gene were significantly expressed (588-up and 608-down) in the N1 group under nitrite stress. Between the N1 and N3 groups, the expression of 1537 genes (751-up and 786-down) were significantly different. KEGG pathway analysis reveals that 11 and 19 pathways were significantly different between N1 and control and between N3 and N1 groups, respectively. Transcriptome results demonstrate that nutrient metabolism is much more active in crabs fed additional selenium under nitrite stress. This study indicates that dietary selenium can improve both antioxidant capacity and immune response and alter the protein and carbohydrate metabolism of E. sinensis under nitrite stress.

Dietary non-protein energy source regulates antioxidant status and immune response of barramundi (Lates calcarifer).

This study evaluates the effects of different dietary sources of non-protein energy on growth performance, histological structure, antioxidant status and immune response of barramundi Lates calcarifer. Fish were fed with isoenergetic diets (18 kJ/g) with two types of non-protein energy in the experimental groups and a regular diet was used as the control for 56 days. The specific growth rate and survival of fish were not significantly different between experimental diets. Hepatic histology did not reveal significant differences between dietary treatments at cellular level. The activity of most antioxidant enzymes in the lipid group significantly increased, and the antioxidant capacity in the carbohydrate group was significantly higher than that in other treatments. In the TOR pathway, LST8 homolog (mLST8) expression in the high lipid group was downregulated, and the mechanistic target of rapamycin (mTOR) expression in the high carbohydrate group was downregulated and eIF4E expression was upregulated. The C-reactive protein (CRP) expression in the high lipid and high carbohydrate groups was upregulated. The expression levels of heat shock protein genes in the high lipid group and the high carbohydrate group were significantly downregulated. This study indicates that the lipid diet have less effect in fish immunity but is more suitable as a non-protein ingredient for energy supply for barramundi.

Effects of glycinin and ß-conglycinin on growth performance and intestinal health in juvenile Chinese mitten crabs (Eriocheir sinensis).

This study investigates the effects of two soybean antigens (glycinin and ß-conglycinin) as an antinutritional substance in the diet on the growth, digestive ability, intestinal health and microbiota of juvenile Chinese mitten crabs (Eriocheir sinensis). The isonitrogenous and isolipidic diets contained two soybean antigens at two levels each (70 and 140 g/kg ß-conglycinin, 80 and 160 g/kg glycinin) and a control diet without ß-conglycinin or glycinin supplementation, and were used respectively to feed juvenile E. sinensis for seven weeks. Dietary inclusion of either glycinin or ß-conglycinin significantly reduced crab survival and weight gain. The crabs fed diets containing soybean antigens had higher malondialdehyde concentrations and lower catalase activities in the intestine than those in the control. The activities of trypsin and amylase in the intestine were suppressed by dietary ß-conglycinin and glycinin. Dietary glycinin or ß-conglycinin impaired the immunity and morphological structure of intestine, especially the peritrophic membrane. The mRNA expression of constitutive and inducible immune responsive genes (lipopolysaccharide-induced TNF-α factor and interleukin-2 enhancer-binding factor 2) increased while the mRNA expression of the main genes related to the structural integrity peritrophic membrane (peritrophin-like gene and peritrophic 2) significantly decreased in the groups with soybean antigen addition. Soybean antigen could also change the intestinal microbial community. The abundance of pathogenic bacteria (Ochrobactrum, Burkholderia and Pseudomonas) increased significantly in both soybean antigen groups. Although pathogenic bacteria Vibrio were up-regulated in the glycinin group, the abundance of Dysgonomonas that degraded lignocellulose and ameliorated the gut environment decreased in the glycinin group. This study indicates that existence of soybean antigens (glycinin or ß-conglycinin) could induce gut inflammation, reshape the community of gut microbiota, and cause digestive dysfunction, ultimately leading to impaired growth in crabs.

Beneficial effects of dietary ß-glucan on growth and health status of Pacific white shrimp Litopenaeus vannamei at low salinity.

An 8-week trial was conducted to evaluate the effect of dietary ß-glucan supplement (0, 0.01%, 0.02%, or 0.04%) on growth and health of Pacific white shrimp Litopenaeus vannamei at low salinity of 3 practical salinity unit (psu). The L. vannamei fed 0.02% and 0.04% ß-glucan gained more weight and showed higher activities of protease, amylase, superoxide dismutase, and glutathione peroxidase in the intestine than in the control (0% ß-glucan). The L. vannamei fed 0.04% ß-glucan had a higher condition factor than those fed the control diet. Amylase activity in the hepatopancreas of L. vannamei fed 0.02% ß-glucan was higher than those fed the control diet. Dietary ß-glucan supplement increased the mRNA expressions of Toll-like receptor, myostatin, immune deficiency or heat shock protein 70, but decreased the mRNA expressions of tumor necrosis factor-α and C-type lectin 3 in both hepatopancreas and intestine. The response of intestine microbiota in L. vannamei fed 0.04% ß-glucan was further compared to the control. The 0.04% ß-glucan supplement reduced richness and diversity of the intestinal microbial community as indicated by the low values of Chao1 estimator, ACE estimator, Simpson index and Shannon diversity index. Abundances of Bacillus, Chitinibacter, Geobacillus and Vibrio in the intestine increased, while Flavobacterium, Microbacterium and Mycobacterium decreased significantly in L. vannamei fed 0.04% ß-glucan compared to the control. This study indicates that dietary ß-glucan supplement at 0.02%-0.04% can significantly improve digestibility, antioxidant capacity and immunity in L. vannamei, and thus improve growth performance and survival at low salinity. These beneficial effects of ß-glucan probably are related to the dominance of probiotics over potential pathogens in the intestine.

Intestinal bacterial signatures of the "cotton shrimp-like" disease explain the change of growth performance and immune responses in Pacific white shrimp (Litopenaeus vannamei).

Imbalance of intestinal microbiota has been recognized in aquatic animals infected with various diseases. However, the signature of intestinal bacteria of the "cotton shrimp-like" disease in the Pacific white shrimp Litopenaeus vannamei remains unknown. This study investigates the composition, diversity, microbial-mediated function and interspecies interaction of intestinal microbiota on shrimp with different health status using 16S rRNA gene high-throughput sequencing. Meanwhile, the growth performance and the mRNA expression of innate immune gene in hepatopancreas were also investigated. The growth performance and the mRNA expression of innate immune genes (e.g., crustin, toll, and immune deficiency genes) in the hepatopancreas were significantly decreased in diseased shrimp compared with healthy shrimp. Bacteria of the family Rickettsiaceae and genus Tenacibaculum were exclusively enriched and significantly increased in diseased shrimp, respectively, whereas, the Actinobacteria class dramatically deceased. The diseased shrimp exhibited higher ACE and Chao1 indices and lower complexity of intestinal interspecies interaction than healthy shrimp. Microbial-mediated functions predicted by PICRUSt showed that 83% KEGG pathway including nutrient absorption and digestion significantly increased in diseased shrimp. This study provides an overview on the interplay among the "cotton shrimp-like" disease, intestinal microbiota, growth performance and host immune responses from an ecological perspective.

Effect of single and combined immunostimulants on growth, anti-oxidation activity, non-specific immunity and resistance to Aeromonas hydrophila in Chinese mitten crab (Eriocheir sinensis).

This study evaluates the effect of dietary supplementation of immunostimulants on the Chinese mitten crab (Eriocheir sinensis) with a single administration of mannan oligosaccharide (MOS), or its combination with either ß-glucan or with inulin for 8 weeks. Four diets included an untreated control diet (C), MOS alone (3 g kg-1, M), MOS with ß-glucan (3 g kg -1 MOS + 1.5 g kg -1 ß-glucan, MB), and MOS with inulin (3 g kg -1 MOS + 10 g kg -1 inulin, MI). The weight gain and specific growth rate of the crabs fed M, MB, and MI diets were improved by lowing feed conversion ratio. The growth and feed utilization of the crabs fed the MB diet were improved compared with the other three groups. The crabs fed the M, MB and MI diets showed a higher intestinal trypsin activity than that in the M and control groups. The highest trypsin activity in the hepatopancreas was observed in the MB group. Crabs fed M, MB and MI diets increased antioxidant system-related enzyme activities, but reduced malondialdehyde. The highest activities of alkaline phosphatase, acid phosphatase, lysozyme and phenol oxidase in the gut and the respiratory burst of the crabs were found in the MB group. The MB diet promoted the mRNA expression of E. sinensis immune genes (ES-PT, ES-Relish, ES-LITAF, p38MAPK and Crustin) compared with the control. After 3 days of infection with Aeromonas hydrophila, the highest survival of crabs was also found in the MB group. This study indicates that the combination of MOS with ß-glucan or with inulin can improve growth, antioxidant capacity, non-specific immunity and disease resistance in E. sinensis.

Arginine supplementation improves growth, antioxidant capacity, immunity and disease resistance of juvenile Chinese mitten crab, Eriocheir sinensis.

To investigate the effects of arginine (Arg) on the growth, antioxidant capacity, immunity and disease resistance of juvenile Chinese mitten crab, three diets containing Arg levels at 1.72% (control), 2.73% and 3.72% were formulated and fed to Chinese mitten crab (0.22 ±â€¯0.03 g) for eight weeks. The weight gain, ecdysterone and growth hormone in the serum, relative expression of insulin-like growth factor 2 in the hepatopancreas significantly increased in crabs fed the 2.73% and 3.72% Arg diets. The protein and lipid contents significantly increased in crabs fed the 3.72% Arg diet. The feed conversion ratios in crabs fed the diets with Arg additions were lower than in the control. Arg supplementation also enhanced the antioxidative capacity by increasing the activities of superoxide dismutase, catalase and the relative expression of Kelch-like ECH-associated protein 1 gene in the hepatopancreas, which subsequently decreased malondialdehyde content in the hepatopancreas. Besides, Arg also decreased nitric oxide content in the serum and the activity of nitric oxide synthetase in the hepatopancreas. The relative mRNA levels of crustin, relish, lysozyme and cryptocyanin genes were significantly upregulated by Arg supplementation. The activities of acid phosphatase and alkaline phosphatase in the serum significantly increased in crabs fed the 3.72% Arg diet than those in the control. Similarly, the relative mRNA levels of crustin, cryptocyanin and proPO genes were significantly upregulated in crabs fed the 2.73% Arg diet after lipopolysaccharide challenge, and in crabs fed the 3.72% Arg diet after the Poly (I:C) challenge. The crabs fed the 2.73% and 3.72% Arg diets had higher survival rate after bacterial infection than those fed the control diet. This study indicates that the addition of Arg to the diet at 2.7-3.7% can improve the growth, survival, antioxidant capacity, immunity and disease resistance in juvenile Chinese mitten crab.

Forskolin reduces fat accumulation in Nile tilapia (Oreochromis niloticus) through stimulating lipolysis and beta-oxidation.

High fat diets are commonly used in aquaculture to reduce feed cost in Nile tilapia, but impair its lipid homeostasis. This study evaluated the role of forskolin on reducing fat accumulation in Nile tilapia (Oreochromis niloticus) by using in vitro and in vivo experiments. The use of 50 µM forskolin in vitro increased free fatty acid and glycerol release, but decreased triglyceride in adipocytes and hepatocytes. The adipose triglyceride lipase (ATGL), protein kinase cAMP-dependent type I regulatory subunit alpha (PKAR I) and other genes related to ß-oxidation (peroxisome proliferator activated receptor alpha, PPARα and carnitine O-palmitoyltransferase 1, CPT1) were significantly up-regulated. After feeding a high-fat diet for six weeks, O. niloticus were fed with 0 (control), 0.5 and 1.5 mg/kg forskolin for two weeks to determine whether forskolin could reduce fat accumulation in vivo. Fish fed the two levels of forskolin decreased significantly the hepatosomatic and mesenteric fat indices. The total lipid in the whole fish and liver together with the serum glycerol content were lower in fish fed on forskolin than in the control. The fish fed on forskolin diets exhibited smaller areas of lipid droplets in adipose and liver tissues. Lipolysis related genes (ATGL, hormone-sensitive lipase, HSL; monoacylglycerol lipase, MGL; and protein kinase cAMP-activated catalytic subunit, PKAC) and ß-oxidation genes (PPARα; fatty acid binding protein 1, FABP1; and CPT1) in the adipose were up-regulated. Similarly, in the liver lipolysis genes such as ATGL and PKAR I and ß-oxidation genes (PPARα, FABP1, CPT1 and acyl-CoA oxidase, ACO) showed an increasing trend with the increase of forskolin doses. This study indicates that forskolin can reduce fat accumulation in the adipose and liver by stimulating lipolysis and ß-oxidation in O. niloticus.

Nutrition and water temperature regulate the expression of heat-shock proteins in golden pompano larvae (Trachinotus ovata, Limmaeus 1758).

Understanding fish larval development is of a great interest for aquaculture production efficiency. Identifying possible indicators of fish larvae stress could improve the production and limit the mortality rate that larval stage is subjected to. Heat-shock proteins (HSPs) and heat-shock factors (HSFs) are well known as indicators of response to many kinds of stressor (e.g., environmental, morphological, or pathological changes). In this study, golden pompano larvae were raised at different temperatures (23 °C, 26 °C, and 29 °C), as well as three different diets (Artemia nauplii unenriched, Artemia nauplii enriched with Nannochloropsis sp., and Artemia nauplii enriched with Algamac 3080), and the expression of HSP60, HSP70, HSF1, HSP2, and GRP94 were monitored. While stress genes were widely expressed in the larval tissues, HSP60 and HSP70 were principally from the gills and heart; HSF1 principally from the muscle, brain, and heart; and GRP94 principally from the head kidney and spleen. Golden pompano larvae were found to be more sensitive to thermal changes at later larval stage, and 29 °C was showed to likely be the best condition for golden pompano larval development. Nannochloropsis sp.-enriched Artemia nauplii treatment was found to be the most appropriate feed type with moderate relative expressions of HSP60, HSP70, HSF1, HSF2, and GRP94.