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Antibacterial properties of cinnamaldehyde against Aeromonas hydrophila were assayed in this study. To investigate the action mechanisms of cinnamaldehyde against A. hydrophila, we examined the antibacterial activity, bacterial membrane permeability, and ultrastructure of A. hydrophila cells treated with cinnamaldehyde. The results showed that the minimum inhibitory concentration (MIC) value of cinnamaldehyde against A. hydrophila NJ-35 was found to be 0.039 mg/mL. The trends of the growth curve of A. hydrophila treated with different concentrations of cinnamaldehyde (from 1/4 MIC to 2 MIC) were different, and there was a significant difference in the growth curve of different groups of treatment. There were significant differences in the K+ concentration among all treatment groups from 1 h to 5 h after incubation compared with that of the control. The highest K+ concentration was observed in the 1 MIC group of cinnamaldehyde. The ultrastructure of A. hydrophila cells treated with cinnamaldehyde was destroyed, and the morphology changed. These results indicated that cinnamaldehyde could inhibit the growth of A. hydrophila, increase bacterial membrane permeability, and damage cell membrane integrity, resulting in leakage of the A. hydrophila cell contents.

We evaluated the effect of dietary supplementation with Moringa oleifera leaf extract on the resistance to Aeromonas hydrophila infection in crucian carp. The fish were randomly divided into five groups: the basal diet, the basal diet supplied with 0.25% (0.25 M), 0.5% (0.5 M), 0.75% (0.75 M) and 1.0% M. oleifera leaf extract (1.0 M) for 8 weeks. The growth, antioxidant capabilities, related immune genes as well as resistance to A. hydrophila infection were determined. The results showed that compared with the control group, the weight gain, specific growth rate in the group of 0.5% M. oleifera leaf extract, serum superoxide dismutase (SOD), albumin (ALB) and glutathione peroxidase (GSH-Px), the gene expression of hepatopancreas BTB and CNC homolog 1 (Bach1), NF-E2-related factor 2 (Nrf2), peroxidases (PRX) and NADPH oxidase (NOX) in the group of 0.5%-1.0% M. oleifera leaf extract increased, while feed conversion ratio, serum cortisol, red blood cell (RBC), alanine aminotransferase (ALT), malonaldehyde (MDA) decreased in the group of 0.5%-1.0% M. oleifera leaf extract before the stress. After the infection, the group of 0.5% or 0.75% M. oleifera leaf extract also could improve the serum ALB, hepatopancreas Kelch-like-ECH-associated protein 1 (Keap1), Bach1, Nrf2, TOR, PRX and NOX and reduce cortisol compared with the control group. In summary, this study suggested that 0.5% M. oleifera leaf extract inclusion increased the growth performance, even had positive effects on physiological and immune function, and enhanced resistance against pathogenic infections in crucian carp. The optimum level of M. oleifera leaf extract for crucian carp was estimated to be 0.35%-0.48% based on polynomial comparison with FCR and SGR.

A 70-day feeding trial was conducted to ascertain the effects of threonine on immune response of juvenile oriental river prawn (Macrobrachium nipponense). Six isonitrogen and isolipidic feeds were formulated according to levels of dietary threonine (0.35%, 0.79%, 1.18%, 1.67%, 2.08% and 2.48% respectively). The juvenile prawns were divided into six groups with four replicates, and stocked into 24 tanks with 50 prawns per tank (initial weight 0.20 ± 0.02 g). The results showed a significant increasing trend of final body weight, specific growth rate, protein efficiency ratio, and weight gain rate when threonine levels increased to 1.67% (P 0.05). However, feed intake, feed conversion ratio, and whole-body lipid composition significantly decreased as threonine levels in the feed increased up to 1.67% (P 0.05). Moreover, haemolymph N-urea content was significantly lowest at 1.67% threonine level (P 0.05), whereas glucose was highest at 0.79% followed by 1.67% of threonine levels in the feeds. Aspartate aminotransferase (AST) enzyme activities were significantly decreased by an imbalance (except 1.67%) of threonine in the feed (P 0.05). Activities of Alanine aminotransferase (ALT) and albumen (ALB) were not significantly affected by threonine in the feed (P 0.05). Excessive dietary threonine level (2.48%) significantly activated haemolymph catalase (CAT) activity (P 0.05), whereas malondialdehyde (MDA) content was significantly affected by deficient (0.35% and 0.79%) dietary threonine levels (P 0.05). Inducible nitric oxide synthase (iNOS) activity and haemolymph complement component 4 (C4) content were significantly decreased by deficient levels of threonine in the feed (P 0.05). Excess threonine concentration significantly down-regulated Toll, Dorsal, Relish, and heat shock protein 60 (Hsp60) gene expressions in the hepatopancreas of M. nipponense (P 0.05), while all genes were significantly up-regulated by the optimal (1.67%) threonine level (P 0.05). The threonine level at which maximum specific growth rate of M. nipponense occurred was estimated by second degree polynomial regression analysis as 1.65% of threonine level, equivalent to 4.44% dry weight bases of protein in the feed.

This study aimed to investigate the effects of dietary tea tree oil (TTO) on the performance, intestinal antioxidant capacity, and non-specific immunity after ammonia nitrogen stress in Macrobrachium rosenbergii. Six experimental diets were formulated with 0, 25, 50, 100, 200, 400 mg/kg TTO, respectively. A total of 900 prawns (average initial weight, 0.39 ± 0.01 g) were randomly assigned to 6 groups in triplicate in 18 tanks. After an 8-week feeding trial, 20 prawns from each tank were changed with 20 mg/L ammonia stress for 24 h. The results showed that 100 mg/kg TTO significantly increased prawns performance and survival rate compared with the control group. Moreover, 100 and 200 mg/kg TTO significantly improved intestinal antioxidant capabilities by increasing SOD enzyme activities and decreasing MDA levels. In addition, the prawns fed with 100 mg/kg TTO diet showed the highest survival rate under ammonia stress. After ammonia stress, the group of 100 mg/kg TTO significantly improved antioxidant capacity by increasing hemolymph respiratory burst activity, as well as intestinal anti-superoxide anion activity and SOD. Coincidentally, 100 mg/kg TTO significantly upregulated the intestinal relative expression of antioxidant-related genes (peroxiredoxin-5). Further, it was found that 100 mg/kg TTO activated the toll-dorsal pathway in prawns, which performed the similar function as the classic NF-κB pathway by upregulating the TNF-α and IL-1. Finally, 100 mg/kg TTO increased the levels of iNOS activities and NO contents after ammonia stress and enhanced non-specific immunity. The results indicated that 100 mg/kg TTO could significantly improve the M. rosenbergii performance, antioxidant capacity and ammonia stress resistance. We suggested that the mechanisms may be attributed to that TTO enhanced the antioxidant capacity and non-specific immunity of M. rosenbergii via the NF-κB signal pathway.

The juvenile carps (Cyprinus carpio) were fed diets with four protein sources (15% and 30% glyphosate-resistant genetically modified (GM) named GM 15 and GM 30, respectively, and 15% and 30% non-genetically modified (NGM) soybean named NGM 15 and NGM 30) for 180 days. Results showed that alkaline phosphatase (ALP) activity for the GM30 group was significantly lower than that of the NGM30 group. The activity of glutathione peroxidase (GSH-Px) in the hepatopancreas of carp for the GM30 group was significantly higher than that of the NGM15 group (P0.05). This study indicates that it may aggravate the damage degree of intestinal epithelial cells of carp and more easily cause liver cell damage in the short term when the amount of GM soybean in the feeds was 30%. Therefore, higher glyphosate-resistant GM soybean may have adverse effects on the carp’s serum, intestinal, and hepatopancreas and considerably reduce the hepatopancreatic carp’s antioxidant capacity.

The use of Clostridium butyricum in crustacean aquaculture for anti-abiotic stress is yet unknown. Feeds were formulated containing 0, 125, 250, 500, and 1000 mg/kg Clostridium butyricum (2 × 10

Lipids are essential nutrients for organisms, and high-fat feeds for shrimp may cause oxidative stress. This study evaluated the effects of feeding high fat in the diet on the growth, antioxidant, immunity, and liver fat accumulation of Macrobrachium rosenbergii post-larvae. Five groups with an initial body weight of 0.0084 ± 0.001 g were fed five isonitrogenous and isoenergetic diets (47.01% crude protein and 18.40 kJ/g gross energy) containing 8%, 10%, 12%, 14% and 16% (named L8, L10, L12, L14 and L16) lipid for 8 weeks, respectively. The results showed that the weight gain rate (WGR) and specific growth rate (SGR) of L8 group were significantly higher than those of L10, L12, L14 and L16 group (P 0.05), and the feed coefficient (FCR) of L8 group was significantly lower than that of other groups (P 0.05). With the increase of dietary fat level, the content of MDA and the activity of SOD increased significantly, and the activities of T-AOC and CAT decreased significantly (P 0.05). HE staining clearly revealed the occurrence of hepatocyte swelling, hepatocyte vacuolization and nucleus displacement to the peripheral cell vacuolization in the L16 group, and hepatic lipid accumulation was further observed in the L14 and L16 group by Oil red O staining. In addition, high-fat diet significantly upregulated the expression of Dorsal, Relish and IκBα mRNA, and also upregulated the expression of fat synthesis-related genes FAS, ACC, DGAT and fat transport-related gene FABP (P 0.05), and significantly downregulated the expression of fat metabolism-related genes AMPK and CPT-1 (P 0.05) compared to that of the L8 group. In conclusion, this study showed that feeding a high-fat diet could induce oxidative stress, inhibit growth performance, alter antioxidant capacity, cause hepatic fat deposition and affect the immune system of M. rosenbergii post-larvae.

This study explored the effects of different hypotonic stress levels on antioxidant capacity, microbial composition, and gene expression of Macrobrachium rosenbergii post-larvae. The salinity of the control group was 15 ‰ (S15), and the hypotonic stress groups included three levels of 10 ‰ (S10), 8 ‰ (S8), and 6 ‰ (S6). Different hypotonic stress levels caused oxidative damage in post-larvae, evidenced by decreased superoxide dismutase (SOD) and anti-superoxide anion free radical (ASAFR). They increased malondialdehyde (MDA), nitric oxide (NO), and inducible nitric oxide synthase (iNOS) levels. Microbiological analysis exhibited that different hypotonic stress levels significantly changed microbial composition and diversity. The microbial composition in the water environment where post-larvae living was different from post-larvae. The pathogenic bacteria, including Vibrio and Flavobacterium, were abundant in S6. Transcriptome analysis showed 2, 7967, 297 DEGs, including 1, 3564, 27 up-regulated genes and 1, 4403, 270 down-regulated genes in S10, S8, and S6 groups, respectively. KEGG enrichment results showed that immune and glucose metabolism-related pathways were enriched significantly. Correlation network analysis demonstrated close interactions among antioxidant parameters, microbes, and differentially-expressed genes. In conclusion, hypotonic stress reduced the antioxidant capacity, caused oxidative damage, and altered microbial composition in M. rosenbergii post-larvae. Moreover, when the salinity is below 8 ‰, hypotonic stress impairs the immune system of M. rosenbergii post-larvae.

A feeding trial was conducted to evaluate the effects of the fishmeal level in the diet of the oriental river prawn, Macrobrachium nipponense, on growth performance and gene expression related to protein and lipid metabolism. Six isonitrogenous (41%) and isolipid (8.5%) diets were designed with decreasing fishmeal (FM) levels from 35% to 30%, 25%, 20%, 15%, and 10%. Each diet was fed to three tanks of 50 prawns for 9 weeks. Decreasing FM levels in the diet from 35 to 25% had no adverse effects on growth performance and feed utilization. However, growth performance and feed utilization were significantly decreased when the FM level in the diet was decreased to below 20%. FM reduction in the diet of oriental river prawn had no effect on survival and whole-body composition with the exception of lipid content. Decreasing the FM level to below 25% upregulated the relative gene expression of TOR, ATF4, and ATF3 in the hepatopancreas. The mRNA levels of ATF4 and ATF3 were upregulated in the muscle of those fed the diet with 25% FM. Reduction of the FM level in the diet to less than 20% increased the relative expression of fatty acid synthesis genes such as FAS, ACC, and G6PDH, and increased the relative gene expression of CPT1. Taken together, a 25% FM level in the diet was found to be optimal for M. nipponense.

The present study was aimed to evaluate the effects of the cyclophosphamide (CY) exposure (Control, 0.032, 0.32, 1.0, 1.6 and 3.2 mg/mL) on the damage in the peripheral blood leukocytes of blunt snout bream for 24 h, which including cell viability, apoptosis, lactate dehydrogenase (LDH) release, mitochondrial membrane potential (Δѱm), ROS, antioxidant enzyme activity and the relative mRNA levels of apoptosis. Results showed that cell viability and Δѱm effects of CY were greatly reduced, and occurred in a dose-dependent manner. CY exposure (0.32–3.2 mg/mL) significantly increased the LDH release and induced apoptosis accompanied by ΔΨm disruption and ROS generation compared to the control. The cellular ROS was significantly increased with increase of CY level from 0.032 mg/mL to 1 mg/mL and the plateau occurred at 0.32 mg/mL. Additionally CY exposure led to oxidative stress as evidenced by significantly the decrease of SOD and CAT and increase of MDA concentration after treating cells with 3.2 mg/mL of CY. Besides, the relative mRNA levels of caspase-3 in the dose of 0.032, 0.32 mg/mL CY, caspase-9 and interleukins-1β (IL-1β) in the dose of 0.32 mg/mL CY, tumor necrosis factor-alpha (TNF-α) in the dose of 0.032 mg/mL CY significantly higher than that of the control. In conclusion, 0.32–3.2 mg/mL CY could lead to cytotoxic effect, inflammatory response and induce the apoptosis of the peripheral blood leukocyte of Megalobrama amblycephala.

Lipids work as essential energy sources for organisms. However, prawns fed on high-fat diets suffer from oxidative stress, whose potential mechanisms are poorly understood. The present study aimed to explore the regulation mechanism of oxidative stress induced by high fat and the amelioration by vitamin E (VE) of oxidative stress.

Agricultural nonpoint source pollution is a critical issue that causes eutrophication in water bodies, due to the application of fertilizers. The traditional rice-fish coculture systems may develop a novel sustainable agriculture. The present study was undertaken to understand the ecological mechanisms of integrated fish farming especially water chemistry, dynamics of plankton, and benthic populations and their community compositions. In the rice-fish coculture system (RF), the total nitrogen in the water was higher compared to the rice monoculture system (RM), but significantly lower than in conventional fertilized rice farming. Dissolved oxygen (DO) presented a significant “Month x Treatment” interaction (p

p65 is one of the important subunits of the inflammation-related transcription factor NF-κB. In the present study, we cloned and identified the p65 from Megalobrama amblycephala (Mnp65) by homologous cloning and RACE technique. The full-length Mnp65 cDNA consisted of 2331 bp, and included one open reading frame encoding a 604-amino acid putative protein. The protein sequence included a DNA binding motif, a well conserved N-terminal Rel-homology domain (RHD), and a C-terminal IG-like plexins transcription (IPT). Mnp65 was closely related with the other p65 proteins of Cypriniformes and clearly distinct from that of Perciformes and Salmoniformes in terms of sequence homology. Mnp65 homodimer may interact with IκBα in the IPT domain based on the predicted 3D structure of IκBα/Mnp65 complex. Mnp65 was ubiquitously expressed in M. amblycephala tissues, and the highest levels were detected in muscle and liver. Intragastric infection with Aeromonas hydrophila caused respiratory burst and cytokine storm from 8 h to 48 h, showing significantly higher level of respiratory burst activities and significantly high cytokines levels, such as TNF-α, IL-1β, IL-6, IL-8 etc., compared to 0 h. In addition, the bacterial challenge downregulated the IkBα, and upregulated Mnp65 and TNF-α in the liver. IkBα-Mnp65 was regulated by the negative feedback of cytokine storm, to increase IkBα and decrease Mnp65. Then cytokine storm was relieved at 96 h. Finally, severe intestinal inflammation was observed from 24 h to 48 h after infection, characterized by extensive villous necrosis, epithelial hyperplasia and lymphocyte infiltration, all of which were relieved at 96 h. Taken together, Mnp65 plays a crucial role in the physiological response of teleost fish to bacterial infection.

In order to study the effects of Moringa oleifera leaf extract on Macrobrachium rosenbergii under high ammonia exposure, freshwater prawns were randomly divided into five groups: a control group was fed with basal diet, and four treatment groups fed with basal diet supplemented with 0.25%, 0.5% and 1.0% M. oleifera leaf extract and 0.025% Enrofloxacin for 60 days, respectively. Then, freshwater prawns were exposed to high ammonia stress for 72 h and Vibro anguillarum infection. The growth, antioxidant capabilities, related immune genes as well as resistance to infection by V. anguillarum were determined. The results showed that compared with the control group, the weight gain, specific growth rate and protein efficiency rate, haemolymph catalase (CAT), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) increased while feed conversion ratio, haemolymph aspartate aminotransferase, alanine aminotransferase, nitrogen oxide (NO), hepatopancreas heat shock proteins (HSP70), immune deficiency (IMD) expression levels decreased in the group of 0.5% M. oleifera leaf extract before the stress. After ammonia stress, the group of 0.5% M. oleifera leaf extract also could improve the haemolymph SOD, glutathione peroxidase, NO, iNOS, hepatopancreas HSP70 expression levels and reduce haemolymph CAT, hepatopancreas peroxiredoxin 5 and NF kappa B inhibitor alpha expression level compared with the control group. The rate of mortality of the prawns challenged with V. anguillarum was lower in the supplemented groups in comparison with the control group with the lowest being in the group of 0.5% M. oleifera leaf extract. Antioxidant activities as well as biochemical parameters in the enrofloxacin group (0.025%E) were not significantly enhanced both pre and post challenge in comparison with the M. oleifera leaf extract groups, showing the superiority of the natural herb over the synthetic antibiotic. In summary, this study suggested that at an inclusion rate of 0.5%, M. oleifera leaf extract could increase the growth performance, even has positive effects on physiological and immune function and prevents high ammonia stress in the Freshwater prawn, M.rosenbergii.

To investigate the regulatory effect of miR-34a on the sirtuin 1(sirt1) / forkhead box O 1 (foxo1) axis in Megalobrama amblycephala under high carbohydrate metabolism, M. amblycephala fish were fed either a control or a high glucose diet for eight weeks. Metabolic models of the high glucose group, normal glucose group, miR-34a inhibition group, and miR-34a overexpression group were constructed using miR-34a inhibitor and miR-34a mimic injection (where appropriate) in M. amblycephala. The results showed that the hepatic expression of SIRT1 was decreased by the high glucose diet after eight weeks, which promoted the expression of foxo1a. Additionally, hepatic pepck and g6pase expression were upregulated, thereby promoting gluconeogenesis. The upregulation of pparα increased the expression of lipid metabolic enzymes such as fas, lps, and acc; therefore, excessive glycogen was deposited as lipid in the adipose tissue. Furthermore, we found that SIRT1 expression increased following miR-34a inhibitor injection, activating foxo1a expression levels, reducing hepatic gluconeogenesis, decreasing hepatic glycogen accumulation, and accelerating hepatic fat decomposition and fatty acid synthesis. In a nutrient metabolic homeostatic state, injection of miR-34a mimics promoted the expression of foxo1a and pepeck, enhanced fat metabolism and fatty acid synthesis, and inhibited SIRT1 protein expression. These results revealed that miR-34a mediates regulation of metabolic enzymes related to hepatic glucose and lipid metabolism by targeting SIRT1/FoxO1 in M. amblycephala under high carbohydrate diet.

This study was aimed to investigate the facilitation of anthraquinone extract on growth performance, immunity, and antioxidant capacity of the oriental river prawn (Macrobrachium nipponense), and whether it could ameliorate the hyperthermia stress. A 12-week rearing experiment was conducted with 0, 125, 250, 500, and 1000 mg kg−1 anthraquinone extract from Rheum officinale Bail-supplemented diets (AE0, AE125, AE250, AE500, and AE1000), and followed a 48-h thermal stress with 32℃ incubation. Results indicate AE250 and AE500 significantly improved the growth performance and feed utilization, the optimum level was estimated to be 251.22 mg kg−1 based on the regression analysis of special growth ratio (SGR). Meanwhile, AE250 and AE500 improved antioxidant enzyme activity and immune-related protein concentration of iNOS-NO signaling. Under thermal stress, AE250 and AE500 improved the heat tolerance, and Toll–Relish signaling was active to the resistance. These results indicate anthraquinone extract could be used as an effective immunostimulant to improve growth performance, physiological balance and protect organism form environmental hyperthermia stress. This may provide insights for immunostimulant development in aquaculture production.

The aroma and taste of Procambarus clarkii (Girard, 1852) fed with different dietary protein were investigated by E-tongue and gas chromatography-ion migration spectrometry (GC-IMS) after cooking. The results showed that dietary protein sources had no significant growth performance. Nevertheless, significantly higher richness taste was observed in animal protein group. The inosine-5'-monophosphate content in animal protein group was significantly higher than that in plant protein group. Twelve aldehydes, eleven alcohols, six ketones, three esters, and two acids were identified in the muscle using GC-IMS. 2-Propanol (monomer), 3-octanol (monomer), 3-furanmethanol (dimer), 2-methyl-1-pentanol (monomer), heptanal (monomer), and allylacetic acid (monomer) changed significantly between dietary animal protein and plant protein. These results suggested that dietary plant and animal protein have a similar effect on the growth performance. For the flavor, the crayfish fed with animal protein had higher volatiles and IMP contents, which might contribute to higher richness.

This study was aimed to investigate the facilitation of anthraquinone extract on growth performance, immunity, and antioxidant capacity of the oriental river prawn (Macrobrachium nipponense), and whether it could ameliorate the hyperthermia stress. A 12-week rearing experiment was conducted with 0, 125, 250, 500, and 1000 mg kg(−1) anthraquinone extract from Rheum officinale Bail-supplemented diets (AE0, AE125, AE250, AE500, and AE1000), and followed a 48-h thermal stress with 32℃ incubation. Results indicate AE250 and AE500 significantly improved the growth performance and feed utilization, the optimum level was estimated to be 251.22 mg kg(−1) based on the regression analysis of special growth ratio (SGR). Meanwhile, AE250 and AE500 improved antioxidant enzyme activity and immune-related protein concentration of iNOS-NO signaling. Under thermal stress, AE250 and AE500 improved the heat tolerance, and Toll–Relish signaling was active to the resistance. These results indicate anthraquinone extract could be used as an effective immunostimulant to improve growth performance, physiological balance and protect organism form environmental hyperthermia stress. This may provide insights for immunostimulant development in aquaculture production.

The study was conducted to evaluate the effects of fish oil replacement by vegetable oils on growth performance, histology, and antioxidant capacity of Macrobrachium rosenbergii. Three isonitrogenous and isoenergetic diets were formulated with different lipid sources included. DFO diet contained 6% fish oil, whereas DSO and DRO diets included 6% soybean oil and rapeseed oil (RO) as alternatives for fish oil, respectively. Prawns were fed thrice daily for 8 weeks. The results showed that prawns in DFO group showed significantly lower final weight, weight gain ratio, and specific growth rate (SGR), but higher feed intake and feed coefficient ratio than those in DSO and DRO groups. In hepatocellular ultrastructure, malformed and atrophic nucleus and higher apoptosis ratio were observed in DFO group. In addition, levels of haemolymph proinflammatory cytokines, activities of anti-superoxide anion, inducible-type NO-synthase (iNOS) and content of nitric oxide, and hepatopancreas NF-κB signal pathway gene expression in DFO group increased markedly compared to those of DSO and DRO groups. The results suggested that vegetable oils, such as soybean oil and RO might be the better lipid sources in diets for Macrobrachium rosenbergii than fish oil, it may be attributed to modified oxidative status induced by NF-κB-NO signal pathway.

Nitrite (NO2−) can cause oxidative stress in aquatic animal when it accumulates in the organism, resulting in different toxic effects on fish. In the present study, we investigated the effects of nitrite exposure on the antioxidant enzymes and glutathione system in the liver of Bighead carp (Aristichthys nobilis). Fish [Initial average weight: (180.05 ± 0.092) g] were exposed to 48.634 mg/L nitrite for 96 h, and a subsequent 96 h for the recovery test. Fish livers were collected to assay antioxidant enzymes activity, hepatic structure and expression of genes after 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 96 h of exposure and12 h, 24 h, 48 h, 72 h, 96 h of recovery. The results showed that the activity of glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), and glutathione reductase (GR) increased significantly in the early stages of nitrite exposure. The study also showed that nitrite significantly up-regulated the mRNA levels of glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), and glutathione reductase (GR) after 6, 48, and 72 h of exposure respectively. Nitrite also increased the formation of malondialdehyde (MDA), oxidized glutathione (GSSG), and the activity of catalase (CAT). Nitrite was observed to reduce the activity of superoxide dismutase (SOD) and the level of glutathione (GSH). In the recovery test, GSH and the GSSG recovered but did not return to pre-stress levels. The results suggested that the glutathione system played important roles in nitrite-induced oxidative stress in fish. The bighead carp responds to oxidative stress by enhancing the activity of GSH-Px, GST, GR and up-regulating the expression level of GSH-Px, GST, GR, a whilst simultaneously maintaining the dynamic balance of GSH/GSSG. CAT was also indispensable. They could reduce the degree of lipid peroxidation, and ultimately protect the body from oxidative damage.

Nitrite (NO2−) contamination of water can severely impact the health of aquatic life and is a major concern for commercial aquaculture. In order to study the effect of nitrite on Aristichthys nobilis, we investigated the oxygen-carrying capacity, NF-κB/HIF-1α pathway, and the gill tissue structure under nitrite stress. In the current study, bighead carp (initial weight 180.05 ± 0.092 g) were exposed to nitrite (48.634 mg/L) for 96 h and then for 96 h recovery test. After nitrite exposure for 6 h, hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) mRNA expression increased significantly in the gill of bighead carp (P

Heat shock proteins (HSPs) play critical roles in the process of anti-stress and immunity and are implicated in autoimmune diseases. In order to understand the comparative stress responses of HSP60 and HSP90β under intermittent thermal stress and Aeromonas hydrophila infection, we cloned their full-length cDNAs from Megalobrama amblycephala liver, predicted their secondary and tertiary structure, and examined their tissue-specific expression patterns. The full length of HSP60 and HSP90β cDNAs indicated that they included all signature sequences of corresponding protein families. They showed high homology to their counterparts in other species, and were consistent with the known classification of fishes based on phylogenetic analysis. HSP60 showed the highest expression in head-kidney, brain, and gill, while HSP90β presented higher in hindgut, liver, and brain. Significant mRNA expression differences were determined between HSP60 and HSP90β in tissues of bladder, liver, heart, and gill. During thermal stress and recovery phase, the highest expression of them were observed at the first recovery for 2 d and 1 d, respectively. The expression between them were extremely significant difference during the first recovery and second stress period. After A. hydrophila infection, their expressions were extremely significantly upregulated. The significant upregulation and rapid response indicated that they were sensitive to thermal stress and bacterial challenge. This study demonstrated that HSP60 and HSP90β might participate in innate immune and environmental responses of M. amblycephala. It indicated that they could be used as biomarkers to test the stress caused by local aquaculture environment.

Microbial colonization is vital for physiological equilibrium in animals. However, the impact of maternal and environmental microbes on microbial succession in the early developmental stages of Macrobrachium rosenbergii remains elusive. In this study, the effects of maternal and environmental microbes on the embryonic and larval microbiota of M. rosenbergii were evaluated by high-throughput sequencing. The results showed that Proteobacteria and Firmicutes were the dominant phyla in the intestine, gonads, and hepatopancreases of maternal prawn. In addition, Actinobacteria was dominant in the intestine while Actinobacteria, Bacteroidetes, and Acidobacteria were dominant in gonads of maternal prawn. During the embryonic stages, Proteobacteria, Actinobacteria, and Bacteroidetes became the dominant phyla. In post-larval stages, Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes tended to dominate. In the water, Proteobacteria, Actinobacteria, and Bacteroidetes were the dominant phyla at 7, 14, and 21 dph water. Maternal microbes prominently impacted the microbial composition during the embryonic stages. Specifically, microbial colonization during embryonic stages was directly related to the maternal hepatopancreas according to source-tracking models. When the post-larvae developed to 7 days, the high contribution to the larval microbiota mimicked the environment. These results indicated that microbial colonization in embryonic and post-larval stages was attributed to the maternal and environmental microbe community, respectively. This study provides a theoretical basis for microbial community manipulation to promote prawn growth and physiological health in aquaculture.

To investigate effects of different temperatures and protein levels on growth performance, physiological response and expression of immune-related genes of M. nipponense. A 3 × 3 factorial design feeding trail was conducted with three levels of water temperature (20 °C, 25 °C and 30 °C) and three levels of dietary protein (P34, P38 and P42) for 8 weeks. The results indicated that weight gain rate (WGR) was significantly affected by water temperature and their interaction. WGR at 25 °C was significantly higher than that at 20 °C and 30 °C. WGR of prawns fed with 38% protein diets was higher than that of the other two treatments at 20 °C. Alanine aminotransferase (ALT) and complement component 4 (C4) were significantly influenced by water temperature and their interaction. Superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and inducible nitric oxide synthase (iNOS) were significantly affected by their interaction. SOD, CAT, MDA and NO were increased with increasing dietary protein level at 20 °C. P42 diets positively increased the levels of SOD, CAT, NO, iNOS and reduced the level of MDA at 25 °C. However, P34 diets positively increased the levels of SOD, CAT, NO and iNOS at 30 °C. The interaction significantly affected Toll, Myd88, Dorsal, IMD, Relish, HSP60 and HSP70 mRNA levels. P38 diets significantly up-regulated the Toll, Myd88, Dorsal, IMD and Relish mRNA levels at 20 °C. Also, P42 diets positively increased their mRNA levels at 25 °C, while negatively down-regulated at 30 °C. Our study indicated that based on WGR, the optimal dietary protein requirement of shrimp is 38%, 42% and 34% at 20 °C, 25 °C and 30 °C, respectively.

The objective of this study was to analyze the target genes and regulatory function of miR-34a in Megalobrama amblycephala using second-generation high-throughput sequencing and bioinformatic tools. Functional enrichment analysis was performed by gene ontology. MiR-34a and target gene expression levels were measured in M. amblycephala fed normal and high-carbohydrate diets. The results revealed that miR-34a was highly conserved in several species, and miR-34a of M. amblycephala has a close evolutionary relationship to that of zebrafish and common carp. miRanda, TargetScan, RNAhybrid predicted 5,185, 6,282 and 2,168 target genes, respectively, and 645 target genes were in common. According to annotation information, the target genes were enriched in phosphate metabolism, glycerophospholipid metabolism, Golgi vesicle transport, cell division, and other biological processes (P

This study evaluated the mechanisms governing insulin resistance, glucose metabolism and lipogenesis in juvenile fish fed with graded levels of dietary arginine. The results showed that, compared with the control group (0.87%), 2.31% dietary arginine level resulted in the upregulation of the relative gene expression of IRS-1, PI3K and Akt in the insulin signaling pathway, while 2.70% dietary arginine level led to inhibition of these genes. 1.62% dietary arginine level upregulated glycolysis by increasing GK mRNA level; 2.70% dietary arginine level upregulated gluconeogenesis and resulted in high plasma glucose content by increasing PEPCK and G6P mRNA level. Furthermore, 2.70% dietary arginine level significantly lowered GLUT2 and increased PK mRNA levels. 1.62% dietary arginine level significantly upregulated ACC, FAS and G6PDH mRNA levels in the fat synthesis pathway and resulted in high plasma TG content. These results indicate that 1.62% dietary arginine level improves glycolysis and fatty acid synthesis in juvenile blunt snout bream. However, 2.70% dietary arginine level results in high plasma glucose, which could lead to negative feedback of insulin resistance, including inhibition of IRS-1 mRNA levels and activation of gluconeogenesis-related gene expression. This mechanism seems to be different from mammals at the molecular level.