Effects of dietary thiamin (vitamin B1) on the growth performance, serum biochemical factors, immune response, and antioxidant activity of great sturgeon (Huso huso) juveniles.

The present research evaluated the positive effects of dietary thiamin (vitamin B1) levels on the growth performance, serum biochemistry factors, immune response, and antioxidant activity of great sturgeon (Huso huso) juveniles. Thiamin was included in diets with levels of 0 (control, T0), 7 (T7), 15 (T15), and 25 (T25) mg/kg diet. Measurements of thiamin levels in diets indicated that they contained 1.80 (T0), 8.02 (T7), 16.2 (T15), and 26.6 (T25) mg thiamin/kg feed. Sturgeon juveniles (240 individuals) with average weight of 44.8 ± 1.96 g were distributed into 12 tanks, and fed with the experimental diets for 8 weeks. Final weight, body weight gain (%), specific growth rate, and feed conversion ratio (FCR) of great sturgeon were significantly influenced by dietary thiamin levels, and the maximum fish performance (P < 0.05) was obtained at a level of 15 mg/kg diet. The trypsin, chymotrypsin, creatine kinase, lipase, α-amylase, and alkaline phosphatase activities were notably (P < 0.05) affected by the dietary thiamin levels. The glucose content was not significantly (P > 0.05) different among the experimental treatments. Diets supplemented with thiamine increased significantly (P < 0.05) triglyceride, cholesterol, and total protein levels accompanied with significant (P < 0.05) decreases in aminotransferase aspartate and alanine aminotransferase activities. Serum antioxidant enzymes were remarkably (P < 0.05) higher, while serum malondialdehyde was significantly (P < 0.05) lower in the thiamin-treated fish compared with the control group. Total immunoglobulin, lysozyme, and ACH50 values were significantly (P < 0.05) higher in fish fed with thiamin-supplemented diets than in the control group. The results of the present study demonstrated that dietary thiamin have an important role in enhancing the growth performance, immune response, and antioxidant activity of great sturgeon. Based on the regression fitting curve of final weight, weight gain, specific growth rate, and FCR values, the optimal level of thiamin is found to be 15.0-17.5 mg/kg diet.

Effects of dietary chitosan oligosaccharide on the growth, intestinal microbiota and immunity of juvenile red claw crayfish (Cherax quadricarinatus).

This study aimed to evaluate the potential benefits of chitosan oligosaccharide (COS) on red claw crayfish (Cherax quadricarinatus) and explore its underlying mechanisms. The crayfish were randomly divided into six groups, and the diets were supplemented with COS at levels of 0 (C0), 0.2 (C1), 0.4 (C2), 0.6 (C3), 0.8 (C4), and 1 (C5) g kg-1. Treatment with COS significantly improved the growth performance of the crayfish with a higher weight gain rate (WGR) and specific growth rate (SGR) in the C2 group compared to the C0 group. Additionally, the content of crude protein in the crayfish muscles in the C1 group was significantly higher than that of the C0 group. Regarding non-specific immunity, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and alkaline phosphatase (AKP), and the levels of expression of the genes related to immunity (SOD; anti-lipopolysaccharide factor [ALF]; thioredoxin1 [Trx1]; C-type lysozyme, [C-LZM]; and GSH-Px) in the hepatopancreas and hemolymph increased significantly (P < 0.05) after supplementation with 0.4 g kg-1 of COS, while the content of malondialdehyde (MDA) decreased (P < 0.05). The survival rate of C. quadricarinatus increased (P < 0.05) in the C2, C3, C4, and C5 groups after the challenge with Aeromonas hydrophila. This study found that COS has the potential to modulate the composition of the intestinal microbiota and significantly reduce the abundance of species of the phylum Proteobacteria and the genera Aeromonas and Vibrio in the gut of C. quadricarinatus, while the abundance of bacteria in the phylum Firmicutes and the genus Candidatus_Hepatoplasma improved significantly. This study suggests that the inclusion of COS in the diet of C. quadricarinatus can enhance growth, boost immunity, and increase resistance to infection with A. hydrophila, especially when supplemented at 0.4-0.8 g kg-1.

Role of dietary tea polyphenols on growth performance and gut health benefits in juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂).

The present study aimed to evaluate the effects of dietary TPs on growth performance, intestinal digestion, microflora and immunity in juvenile hybrid sturgeon. A total of 450 fish (97.20 ± 0.18 g) were randomly divided into a standard diet (TP-0) or four treatments consisting of a standard diet supplemented with four concentrations of TPs (mg/kg): 100 (TP-100), 300 (TP-300), 500 (TP-500), and 1000 (TP-1000) for 56 days. The TP-300 significantly increased weight gain rate (WGR) and specific growth rate (SGR) (p < 0.05), and TP-1000 significantly increased the feed conversion ratio (FCR) (p < 0.05). TP-300 and TP-500 significantly increased intestinal trypsin, amylase, and lipase activities (p < 0.05). Besides, TP-300 significantly enhanced total antioxidant capacity (T-AOC) and the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) and decreased malondialdehyde (MDA) content (p < 0.05). Moreover, TP-300 decreased the expression levels of tumor necrosis factor-alpha (TNF-α), interleukin 8 (IL-8), and interleukin 1ß(IL-1ß) compared with TP-0 and TP-1000 (p < 0.05). In addition, the intestinal microbiota diversity in the TP-300 group was observably higher, the dominant microbiota was Bacteroidota, Cyanobacteria, Proteobacteria and Firmicutes at the phylum level, Enterobacteriaceae, Nostocaceae and Clostridiaceae at the family level. The relative abundances of potential probiotics including Rhodobacteraceae and potential pathogens especially Clostridiaceae were the highest, and lowest, respectively. In conclusion, TP-300 altered the abundance of microbial taxa, resulting in enhancing the intestinal digestion, antioxidant status and non-specific immunity to improve the growth performance in juvenile hybrid sturgeon.

Effect of dietary Bacillus subtilis supplement on Cd toxicokinetics and Cd-induced immune and antioxidant impairment of Procambarus clarkii.

Cadmium (Cd), a non-biodegradable contaminant in freshwater ecosystems, can pose a serious threat to aquatic animals at high levels. In this study, the Cd toxicokinetics and the immune and antioxidant defense were explored in Procambarus clarkii exposed to different levels of Cd (0, 0.1, 1.0 mg Cd/L) or treated with 1.0 mg Cd/L and dietary Bacillus subtilis supplementation (1 × 107 cfu/g). Results from the 21-day uptake and depuration experiment revealed that Cd exposure elicited a dose- and time-dependent uptake in all crayfish tissues, and the rank order of Cd concentration was gill > hepatopancreas > exoskeleton > muscle. The one-compartment model demonstrated that gills had the highest uptake rate (ku) value after Cd aqueous exposure and the ku and elimination rate (kd) values in gill, hepatopancreas, and exoskeleton of the group with 1.0 mg Cd/L were higher than those of the group at alow Cd concentration (0.1 mg Cd/L). However, B. subtilis could decrease Cd ku and increase Cd kd in hepatopancreas, resulting in the reduction of bioconcentration factors (BCF), steady-state concentrations (Css), and biological half-life (Tb1/2). A positive correlation was found between aqueous Cd concentration and the severity of hepatopancreas histopathological injury, while B. subtilis could ameliorate the pathological damage in the high Cd group. Similarly, aqueous exposure to Cd elevated malonaldehyde (MDA) content and suppressed the activities of lysozyme (LZM), acid phosphatase (ACP) in hepatopancreas and alkaline phosphatase (AKP) in hemolymph. The activities of superoxide dismutase (SOD) and catalase (CAT) in hepatopancreas were also inhibited. Nevertheless, they were all recovered with the dietary addition of B. subtilis. In conclusion, our results indicated that exposure to Cd significantly increased Cd accumulation and toxic damages in crayfish hepatopancreas, while dietary administration of B. subtilis to crayfish significantly decreased Cd accumulation and improved the immune and antioxidant defense, leading to the prevention in toxic effects of Cd.

Traditional Chinese herb formulas in diet enhance the non-specific immune responses of yellow catfish (Pelteobagrus fulvidraco) and resistance against Aeromonas hydrophila.

The effects of a traditional Chinese herbal mixture (TCHM) composed of Glycyrrhiza uralensis, Astragalus membranaceus, Rheum palmatum, Catsia tora and Lonicera japonica on immune response and disease resistance of yellow catfish (Pelteobagrus fulvidraco) were studied. Fish were fed diets containing 0% (control), 1.0%, 3.0% or 5.0% TCHM (w/w) for 28 d. Immune parameters including cytokine genes interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and Immunoglobulin M (IgM), acid phosphatase (ACP), alkaline phosphatase (AKP), lysozyme (LZM), catalase (CAT), superoxide dismutase (SOD) and immunoglobulin M (IgM) were measured during the test period. After 28 d of feeding, fish were infected with Aeromonas hydrophila, and mortality was recorded. The TCHM-supplementation diet stimulated ACP, AKP, LZM, CAT, SOD, and IgM activity in serum and induced IL-1ß, TNF-α, and IgM mRNA expression in the spleen. All TCHM groups showed reduced mortality after A. hydrophila infection compared to the control group. These results suggest that the TCHM-supplemented diet can improve fish immunity and disease resistance against A. hydrophila.

Effects of dietary tea tree oil on the growth, physiological and non-specific immunity response in the giant freshwater prawn (Macrobrachium rosenbergii) under high ammonia stress.

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.

Effects of dietary trehalose on growth, trehalose content, non-specific immunity, gene expression and desiccation resistance of juvenile red claw crayfish (Cherax quadricarinatus).

This study was performed to investigate the effects of dietary trehalose on growth, muscle composition, non-specific immune responses, gene expression and desiccation resistance of juvenile red claw crayfish (Cherax quadricarinatus). A total of 540 (body weight of 0.41 ± 0.05) crayfish were randomly divided into six groups for a feeding experiment. Six diets with trehalose levels at 0 (Diet 1), 1 (Diet 2), 2 (Diet 3), 5 (Diet 4), 10 (Diet 5) and 15 (Diet 6) g kg-1 were prepared to feed juvenile red claw crayfish for 8 weeks. The results showed that the weight gain rate (WGR) and specific growth rate (SGR) of crayfish in Diet 4, Diet 5 and Diet 6 groups were significantly improved compared with the control group (Diet 1). Muscle crude protein contents of crayfish fed Diet 4, Diet 5 and Diet 6 were significantly higher than those of the control group. The activities of superoxide dismutase (SOD) and alkaline phosphatase (AKP) in hepatopancreas and hemolymph of crayfish for Diet 4, Diet 5, and Diet 6 groups were significantly increased while malondialdehyde (MDA) content was significantly reduced when compared with the control. The total antioxidant capacity (T-AOC), catalase (CAT) and glutathione peroxidase (GPx) activities in the hepatopancreas and hemolymph of crayfish fed Diet 5 and Diet 6 were significantly higher than those in the control group. However, acid phosphatase (ACP) activity was not significantly different among all experimental groups. The hepatopancreas and intestine trehalose contents of crayfish showed an upward trend with the increase of dietary trehalose levels. Compared with the control group, supplementation of 5-15 g kg-1 trehalose in the feed up-regulated the expression levels of GPx, C-type lysozyme (C-LZM), antilipolysacchride factor (ALF), facilitated trehalose transporter homolog isoform X2 (Tret1-2) and facilitated trehalose transporter isoform X4 (Tret1-4) mRNA. In addition, supplementation of 5-15 g kg-1 trehalose in the feed could improve the survival rate of red claw crayfish under desiccation stress. These results suggested that supplementation of 5-15 g kg-1 trehalose in feed could significantly improve the growth performance, muscle protein, non-specific immunity and desiccation resistance of juvenile red claw crayfish.

Therapeutic effect and immune mechanism of chitosan-gentamicin conjugate on Pacific white shrimp (Litopenaeus vannamei) infected with Vibrio parahaemolyticus.

To explore the disease resistance mechanism of chitosan conjugates, chitosan-gentamicin conjugate (CS-GT) was synthesized and systematically characterized, the immune mechanism of CS-GT on Litopenaeus vannamei infected with Vibrio parahaemolyticus was further explored. The results showed that imine groups in CS-GT were effectively reduced. Dietary supplementation of CS-GT can significantly increase the survival rate, total hemocyte counts, the antioxidant and immune related enzyme activity levels of shrimps (P < 0.05), which are all dose-dependent under the experimental conditions. In addition, CS-GT can protect the hepatopancreas from invading bacteria and alleviate inflammation. Particularly, CS-GT promotes the expressions of legumain (LGMN), lysosomal acid lipase (LIPA) and Niemann-Pick type C2 (NPC2) up-regulated. It is speculated that CS-GT may stimulate the lysosome to phagocytose pathogens more effectively. In conclusions, shrimps fed with CS-GT can produce immune response via lysosome and greatly improve the disease resistance to Vibrio parahaemolyticus.

Dietary ß-glucan (MacroGard®) improves innate immune responses and disease resistance in Nile tilapia regardless of the administration period.

The effects of dietary ß-glucan on innate immune responses have been shown in a number of different vertebrate species. However, there is conflicting information about the period of administration (shorter vs. longer), and it is also unclear to what extent ß-glucan's effects can be observed post-treatment in fish. Thus, we fed Nile tilapia for 0 (control group; 45 days of control diet), 15 (30 days of control followed by 15 days of ß-glucan), 30 (15 days of control followed by 30 days of ß-glucan) or 45 days with a diet containing 0.1% of ß-glucan (MacroGard®). We evaluated the growth performance at the end of the ß-glucan feeding trial and the innate immune function immediately after the feeding trial and 7 and 14 days post-feeding trial. In addition, at day 10 post-feeding trial, we assessed the tilapia's resistance against a bacterial infection. No significant differences were observed in growth performance between the groups; however, fish fed with ß-glucan for 30 and 45 days had higher (approx. 8%) relative weight gain compared to the control. Regardless of the administration period, fish fed with ß-glucan had higher innate immune responses immediately after the feeding trial such as lysozyme activity in plasma, liver and intestine and respiratory burst compared to the control, and in general these differences were gradually reduced over the withdrawal period (up to 14 days). No differences were observed in the plasma hemolytic activity of the complement or myeloperoxidase activity in plasma or intestine. Moreover, fish from the control group had early mortalities (2 vs. 4-5 days post-infection, respectively) and a lower survival rate (60 vs. 80%, respectively) compared to fish fed with ß-glucan for 15 or 30 days, and, interestingly, fish fed for 45 days with ß-glucan had no mortality. This study indicates that regardless of the administration period (i.e., 15 up to 45 days), the ß-glucan improved the innate immune responses and the tilapia's resistance to disease, and this protection could be observed up to 10 days post-feeding trial, adding in vivo evidence that ß-glucan may contribute to a trained innate immunity. Additionally, we showed that a longer period of administration did not cause immunosuppression as previously hypothesized but promoted further growth and immune performance. These findings are relevant to the aquaculture industry and demonstrate that a longer ß-glucan feeding protocol may be considered to achieve better results.

Black soldier fly (Hermetia illucens) larvae meal in diets of European seabass: Effects on antioxidative capacity, non-specific immunity, transcriptomic responses, and resistance to the challenge with Vibrio alginolyticus.

Black soldier fly larvae meal (BSFLM) has been successfully demonstrated as a promising fish meal (FM) replacer in diets of several fish species including European seabass (Dicentrarchus labrax). However, its impacts on antioxidant capacity, and immune responses of treated fish are still poorly understood. A 60-day feeding trial was carried out to evaluate the effects of partial substitution of FM with different levels of dry BSFLM on the antioxidative status, non-specific immunity, transcriptomic responses, and resistance of European seabass to the challenge with Vibrio alginolyticus. Four isoproteic (45%) and isolipidic diets were formulated by replacing 0.0%, 25%, 35%, and 50% of the dietary FM. Each diet was randomly assigned to four fish groups (in triplicates) (initial mean body weight, 12.1 ± 0.21 g) (20 fish per aquarium) (n = 240). Fish were fed three times daily to the apparent satiation. At the end of the feeding trial, serum antioxidant biomarkers such as malondialdehyde levels, and catalase, superoxide dismutase, and glutathione peroxidase enzyme activities were significantly increased in all BSFLM groups in a dose-dependent manner compared to the control group (P < 0.05). The non-specific immune indices, including phagocytic activity, phagocytic index, serum lysozyme and respiratory burst activities were significantly elevated in BSFLM groups compared to those in the control group (P < 0.05). Significant upregulation of the mRNA expression levels of hepatic heat shock protein 70, interleukin-1beta and interleukin-10 genes were observed in all BSFLM groups compared to the control group (P < 0.05). Additionally, after the challenge with V. alginolyticus, the relative percent of survival was significantly elevated in fish groups fed on diets containing graded levels of BSFLM over the control group (P < 0.05). Conclusively, the present study suggests the potential efficacy of partial replacement of dietary FM protein for up to 50% by BSFLM without negative effects on fish health with possible potentiation of the antioxidative status, and the immune responses of the European seabass.

The Synergistic Effects of Combined Use of Mentha longifolia, Thymus carmanicus, and Trachyspermum copticum on Growth Performance, Feed Utilization, and Expression of Key Immune Genes in Rainbow Trout (Oncorhynchus mykiss).

Medicinal plants exhibit remarkable positive effects on different aspects of fish physiology. This study aimed to evaluate the possible impact of a combination of plants (Mentha longifolia, Thymus carmanicus, and Trachyspermum copticum) on growth performance, immune responses and key immune gene expression of rainbow trout. For this purpose, four diets were designed, including zero, 0.25, 0.5, and 1% of a mixture of plants per kg of diet, representing dietary treatments of control, T1, T2, and T3, respectively. Two hundred forty fish (weighing 23.11 ± 0.57 g) were fed 3% of body weight twice a day for 45 days. The results showed that growth parameters of weight gain (except for T1) and FCR were significantly improved in fish receiving all levels of plants, with T3 showing the best growth results. Digestive enzymes activities were notably increased in T1 and T2 compared to the control. Stress biomarkers (glucose and cortisol) were significantly decreased in T1 and T2, while T3 was not significantly different from the control. Immunological responses were significantly improved in T2, while T1 andT3 did not show a statistical difference in terms of lysozyme activity. Catalase activity was noticeably decreased in T1, although superoxide dismutase and malondialdehyde were highest in T2. Immune-related genes were significantly up-regulated in T3 compared to other treatments. Also, antioxidant enzyme coding genes were strongly up-regulated in T2 and T3. Overall, the present results suggest that 1% inclusion of the mixture of M. longifolia, T. carmanicus, and T. copticum (T2) can be used to improve the growth and immunity of rainbow trout.

Micro-algal astaxanthin could improve the antioxidant capability, immunity and ammonia resistance of juvenile Chinese mitten crab, Eriocheir sinensis.

Green alga Haematococcus pluvialis is an important source of natural astaxanthin (Ast), which have been shown to be beneficial for the color formulation, survival, antioxidation, immunity and stress resistance of many crustacean. This study was conducted to investigate the effects of dietary supplementation of H. pluvialis meal on growth, antioxidant status, ammonia resistance, color parameters, and carotenoids composition of juvenile Chinese mitten crab Eriocheir sinensis. Five diets were formulated to contain 0, 30, 60, 90 and 120 mg/kg dry diets of natural Ast (defined as Diet 1-5) using H. pluvialis meal as astaxanthin source. The results showed that: (1) Although all treatments with Ast supplementation had the relatively higher growth performance and survival than the control (Diet 1 treatment), no significant differences were found on growth performance, feed conversion ratio and hepatosomatic index among all treatments. (2) The highest total antioxidant capacity (T-AOC) in hepatopancreas and hemolymph were observed in Diet 4 and 3 treatments respectively, while the lowest malondialdehyde (MDA) contents in hepatopancreas and hemolymph were also found in these two treatments. Furthermore, the significantly positive relationships were detected on acid phosphatase (ACP) activities and dietary Ast contents for hepatopancreas and hemolymph. (3) Diet 3 treatment had the highest mRNA levels of EsLecA, EsTrx, and EsPrx6 in hepatopancreas, while both Diet 3 and 4 treatments reached the peaks for mRNA expression levels of EsMyd88 and EsHc, respectively. (4) The stress test with ammonia-N indicated Diet 1 treatment had the highest mortality among all treatments, and the lowest mortality was found on Diet 3 treatment during the stress test. (5) Dietary Ast significantly improved the redness (a*) of carapace and hepatopancreas, which were consistent with the Ast contents in these tissues from the different treatments. Ast concentrations in carapace reached the plateau for Diet 3 treatment while hepatopancreatic Ast concentration kept increasing with elevating dietary Ast contents. In conclusion, natural astaxanthin could enhance the antioxidative capability, non-specific immunity, tissue Ast contents and stress resistance to ammonia-N, and these results suggested the optimal diet micro-algal astaxanthin was around 60 mg/kg for juvenile E. sinensis.

Modulation of growth performance, non-specific immunity, intestinal morphology, the response to hypoxia stress and resistance to Aeromonas hydrophila of grass carp (Ctenopharyngodon idella) by dietary supplementation of a multi-strain probiotic.

The present study was conducted to evaluate a multi-strain probiotic (MP) on growth performance, immune and antioxidant function, response to hypoxia stress and resistance to Aeromonas hydrophila of grass carp (Ctenopharyngodon idella). Based on the viable cell counts of aerobic Bacillus spp., six experimental diets with MP supplemented at 0, 0.34, 1.68, 3.36, 6.72, 10.1 g kg-1 were formulated and 900 juveniles (7.30 ± 0.01 g) were equally distributed into 30 aquaria with respective diet for 60 days. Results showed that fish with 0.34-1.68 g kg-1 MP had better growth and feed utilization. Further, plasma total protein, albumin and high-density lipoprotein were remarkably increased with dietary MP at >1.68 g kg-1. Dietary MP supplementation at 6.72-10.1 g kg-1 strikingly elevated plasma myeloperoxidase activity and complement C3 content. For fish with MP at 1.68 and 6.72-10.1 g kg-1, their liver malondialdehyde and glutathione peroxidase were remarkably declined and promoted. After hypoxia stress, fish with 3.36-6.72 g kg-1 MP showed significantly higher respiratory burst activity. Challenge test by A. hydrophila confirmed the protection effects of MP through the decreased cumulative mortality rates. For intestinal histomorphology and enzymatic analyses, fish with 1.68 g kg-1 MP displayed significantly higher intestinal villi height, goblet cells and alkaline phosphatase activity. In conclusion, dietary MP supplementation at 1.68 g kg-1 could promote growth, intestinal morphology and antioxidant capacity, while enhancing host immunity requires higher dosages of MP. Broken-line analysis of weight gain revealed that 1.34 g kg-1 is the optimum dosage for the growth of grass carp.

Effects of dietary n-3 highly unsaturated fatty acids on growth, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu).

This study was conducted to investigate the effects of dietary n-3 highly unsaturated fatty acids (n-3 HUFA) on growth performance, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu). Six isoproteic and isolipidic experimental diets were formulated with graded levels of n-3 HUFA (0.65, 1.00, 1.35, 1.70, 2.05 and 2.40% of dry matter, respectively), and the 0.65% group was used as control group. Each diet was randomly allocated to triplicate groups of fish in 1000 L fiberglass tank, and each tank was stocked with 40 fish (initial weight 12.06 ± 0.01 g) for 8 weeks. Results showed that feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI) and condition factor (CF) were all not significantly affected by dietary n-3 HUFA levels (P > 0.05). Weight gain (WG) and specific growth rate (SGR) in 1.35% group were significantly higher than those in 2.40% group (P < 0.05). Crude lipid of body in 1.00% group was significantly lower than that in 1.70% and 2.40% groups (P < 0.05). Liver and muscle fatty acid profiles reflected that of diets. Before challenge with Vibrio harveyi, the activity of serum superoxide dismutase (SOD), catalase (CAT) and content of complement 3 (C3) in 1.35% and 1.70% groups significantly higher than those of control group (P < 0.05). After challenge with Vibrio harveyi, serum CAT, glutathione peroxidase (GSH-PX), lysozyme (LZM) and C3 all increased sharply, while SOD showed the opposite trend. Before challenge with Vibrio harveyi, the expression levels of intestine toll-like receptor 22 (TLR22) and myeloid differentiation factor 88 (MyD88) mRNA in 2.40% group were significantly increased, and the expression levels of tumour necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) mRNA in 2.05% group were significantly higher than those in 1.00% and 1.35% groups (P < 0.05). In addition, the TLR22 and IL-1ß mRNA levels in kidney of 1.70% group were significantly lower than those in control group (P < 0.05). After challenge with Vibrio harveyi, the expression level of MyD88 mRNA in intestine of 1.35% group was significantly higher than that in 1.00% group and from 1.70% to 2.40% groups (P < 0.05), while TNF-α and IL-1ß obtained minimum values in 1.70% group. In the kidney, the interleukin 10 (IL10) mRNA expression was significantly higher in 1.70% group than that in other groups, while the IL-1ß expression in 1.70% group was on the contrary and significantly lower than that in 2.40% group (P < 0.05). Results of this study suggested that moderate dietary n-3 HUFA (1.47%-1.70% HUFA) could improve the growth performance, non-specific immunity and inhibit the inflammatory response of hybrid grouper.

Non-specific immunity promotion in response to garlic extract supplemented diets in female Guppy (Poecilia reticulata).

In the present study, the effect of aqueous garlic (Allium sativum) extract on skin mucus immune parameters of Poecilia reticulata was evaluated. A total of 240 P. reticulate juveniles, weighing 0.013 ± 0.001 g, were randomly stocked in 12 experimental glass tanks at a density of 30 fish per tank. The fish were fed with diets supplemented with garlic extract at the concentrations of 0, 0.10, 0.15, and 0.20 ml kg-1 of diet three times a day at a ratio of 2.5% of their body weight for 80 days. Results of the study showed that, lysozyme activity increased significantly in the skin mucus of garlic-treated Guppy (P < 0.05). Values of ACH50 significantly increased (P < 0.05) in fish by increasing concentration of garlic extract from 0 to 0.15 ml kg-1 in diet. ACH50 also showed a significant declining trend (P < 0.05) by increasing concentration of garlic extract from 0.15 to 0.20 ml kg-1. Total Ig content was enhanced significantly in 0.15 and 0.20 ml kg-1 treatments as well. ALP activity significantly increased in garlic-fed treatments compared to the control. Inclusion of garlic extract in diet was found to have no significant effect on final body weight and weight gain. Results revealed that, administration of 0.15 mL of garlic extract per kg feed is suggested to obtain optimal skin mucus immunity in P. reticulata.

Study on Schizochytrium sp. improving the growth performance and non-specific immunity of golden pompano (Trachinotus ovatus) while not affecting the antioxidant capacity.

A feeding experiment was conducted to determine the effects of Schizochytrium sp. on growth performance, antioxidant capacity and non-specific immunity in golden pompano (Trachinotus ovatus).Two diets were formulated with or without Schizochytrium sp. supplemented (D1:0% and D2: 3%) to feed fish for 8 weeks. Results showed that growth performance, feed intake and survival rate increased significantly with Schizochytrium sp. supplemented (P < 0.05). Feed coefficient rate (FCR) of golden pompano fed the diet supplemented with Schizochytrium sp. was significantly lower than that of fish fed the control diet (P < 0.05). No significant differences were found in antioxidant capacity both in transcriptional level, including nclear factor erythroid-2-related factor-2 (Nrf2), Kelch-like-ECH-associated protein (keap1), catalase (CAT), glutathione peroxidase (GSH-PX) and heme oxygenase 1 (HO-1) and enzyme activity, such as total antioxidant capacity (T-AOC), malondialdehyde (MDA) and superoxide dismutase (SOD) (P > 0.05). Gut amylase and lipase were significantly higher in dietary Schizochytrium sp. supplemented treatment than that in control group (P < 0.05). The relative peroxisome proliferator-activated receptor-α (PPARα) expression level in liver was significantly higher in Schizochytrium sp supplemented treatment than that in control one (P < 0.05). The mRNA expression of myeloid differentiation factor 88 (MyD88), IL-1R-associated kinases 4 (IRAK4), interferon regulating Factor 3 (IRF3), interferon regulating Factor 3(IRF7) and heat shock protein 70 (HSP70) were significantly lower in Schizochytrium sp. supplemented treatment than that in control one (P < 0.05). In Schizochytrium sp. supplemented diet, golden pompano had significantly longer villi length than that in control diet (P < 0.05); muscle thickness in Schizochytrium sp. supplemented diet was thicker than that in control one (P < 0.05) and there were more goblet cells in Schizochytrium sp. treatment (P < 0.05). After the rearing trial, an air exposure trial was conducted. Results showed that the air-exposure mortality (AEM) and mRNA expression level of Nrf2, keap1, CAT, GSH-PX and HO-1 showed no significant difference (P > 0.05). These results indicated that dietary Schizochytrium sp. improved the growth performance and non-specific immunity of golden pompano while made no difference to antioxidant capacity.

Effects of dietary supplementation with icariin on growth performance, antioxidant capacity and non-specific immunity of Chinese mitten crab (Eriocheir sinensis).

We investigated the effects of icariin (ICA) on growth performance, antioxidant capacity and non-specific immunity in Chinese mitten crab (Eriocheir sinensis). A total of 200 healthy crabs (average weight: 33.58 ±â€¯0.05 g) were randomly assigned to four treatments with five replicates, each with ten individuals per pool. There were four dietary treatments: the control group (fed with the basal diet), the ICA 50 group, the ICA100 group, and the ICA 200 group (fed with the basal diet supplemented with 50, 100, and 200 mg/kg ICA, respectively). These diets were provided for 8 weeks. Results indicated that ICA100 crabs had higher weight gain (WG), specific growth rate (SGR) and survival rate (SR) than the controls. Protein carbonyl content (PCC) and malondialdehyde (MDA) concentrations in the haemolymph and hepatopancreas of ICA100 crabs were significantly lower than in the control group, while the superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were significantly higher. The activities of PO, LZM, ACP and AKP were significantly enhanced with ICA supplementation at 50 and 100 mg/kg, yet decreased subsequently at 200 mg/kg. Furthermore, supplementation of 100 mg/kg ICA up-regulated the mRNA expression of prophenoloxidase (proPO), catalase (CAT), mitochondrial manganese superoxide dismutase (mtMnSOD), thioredoxin-1 (Trx1) and peroxiredoxin 6 (Prx6), while the mRNA expression of toll like receptors (TLRs), NF-κB-like transcription factor Relish and lipopolysaccharide-induced TNF-α factor (LITAF) were down-regulated in the hepatopancreas (P < 0.05). These findings indicate that dietary ICA supplementation at an optimum dose of 100 mg/kg may be effective in improving growth performance, antioxidant capability and non-specific immunity of Chinese mitten crab.

Polysaccharide fraction from the Indian mistletoe, Dendrophthoe falcata (L.f.) Ettingsh enhances innate immunity and disease resistance in Oreochromis niloticus (Linn.).

The polysaccharide fraction (PF) isolated from the hemiparasitic mistletoe, Dendrophthoe falcata (L.f.) Ettingsh (DF) leaves was tested for its immunostimulatory properties in Oreochromis niloticus (Linn.). Different groups of experimental fish were fed for 1, 2 or 3 weeks with three different doses [low (0.01%), mid (0.1%), or high (1%)] of D. falcata polysaccharide fraction (DFPF) - supplemented diet. After every feeding regimen, the fish were assessed for non-specific immunological parameters, immune related gene expression and disease protection. The DFPF treated groups showed significant (P < 0.05) enhancement of non-specific immune parameters. Significant (P < 0.05) upregulation of lysozyme and TNF-α gene expression was observed in DFPF treated groups. In pathogen challenge studies using Aeromonas hydrophila, the DFPF treated groups displayed significant (P < 0.05) decrease in percentage mortality and the consequent increase in relative percent survival (RPS). Supplementation of 1% DFPF in the feed for a week conferred the maximum protection against the virulent pathogen challenge, recording a RPS of 100. These results suggest that DFPF has the potential to be used as an immunostimulating feed additive in finfish aquaculture.

Dietary L-Tryptophan potentiates non-specific immunity in Labeo rohita fingerlings reared under elevated temperature.

At present environmental scenario global climate change is a reality and its affect all living organism including fish. The aquatic ecosystem is the most affected system as it is the biggest sink for global warming and elevated temperature and obviously affects all the aquatic life forms. With this hypothesis an experiment was conducted to evaluate the effect of elevated temperature on Labeo rohita fingerlings and potential of dietary L-tryptophan (TRP) in mitigating the effects elevated temperature and enhancing the non-specific immunity. Seven hundred and twenty fishes were randomly distributed in three different thermal groups each with three replicates. The thermal groups were ambient temperature (26 °C), 34 and 38 °C. Then each thermal groups were fed with four different formulated diets containing 0. 0.36%, 0.72% and 1.44% TRP. The effect of dietary TRP supplementation was studied on stress responses, such as cortisol, blood glucose, histopathological changes in liver and kidney and immuno-hematological changes such as red blood cell count (RBC), haemoglobin (Hb), white blood cell count (WBC), lysozyme, nitroblue tetrazolium (NBT), total serum protein, albumin, globulin and albumin-globulin ratio. Subsequently the treated fish were subjected to challenge test with Aeromonas hydrophila. In the present study, primary stress markers were noticeably (p < 0.01) elevated with temperature stress and levels were reduced with nutritional supplementation of TRP. Similarly, immuno-hematological parameters were altered with the exposure of temeparture stress and got improved with dietary TRP supplementation. Results of the present study suggest that dietary supplementation of 1.44% tryptophan has definitive role in the mitigation of temperature stress and gives protection against bacterial infection to L. rohita.

Effects of alpha-ketoglutarate (AKG) on growth performance and non-specific immunity of juvenile red drum fed diets with low or adequate phosphorus levels.

This study evaluated the effects of α-ketoglutarate (AKG) on the growth performance, body, and skeletal composition, as well as non-specific immunity of juvenile red drum (Sciaenops ocellatus) fed adequate or low-phosphorus diets. A 2 × 2 factorial design was arranged with two levels of total phosphorus (1.0 or 1.6%) and two levels (0 or 1%) of AKG. Each diet was fed to red drum in four replicate 110-L aquaria (15 fish/aquarium) twice daily for 7 weeks. The results indicated that the low-P diets significantly (P < 0.05) decreased weight gain, feed efficiency, and condition factor of red drum, but increased moisture content and decreased ash content of whole body, as well as decreased ash content of skeletal tissue and decreased plasma lysozyme. Supplementation of AKG significantly improved the fillet yield and plasma lysozyme and tended to improve neutrophil oxidative radical production (P = 0.097). Based on these results, phosphorus level had the greatest effect on growth performance and non-specific immunity of red drum, while AKG supplementation had limited positive effects on immunological responses and fillet yield of juvenile red drum.