Effects of bioflocs with different C/N ratios on growth,immunological parameters,antioxidants and culture water quality in Opsariichthys kaopingensis Dybowski

Biofloc technology (BFT) is a new ecological aquaculture technology that is intended not only to eradicate pollutants and elevate feed utilization but also to enhance immunity and antioxidant activity in aquatic animals. A 28‐day feeding trial was implemented to evaluate the effects of promoted bioflocs on the water quality, growth performance, immunological parameters and antioxidant status of Opsariichthys kaopingensis Dybowski juveniles in low exchange culture tanks. Three hundred sixty healthy O. kaopingensis juveniles (7.1 ± 0.02 g) were irregularly distributed among 12 tanks. Four C:N ratios in triplicate tanks were tried: C/N = 10.8:1 with a commercial diet (control), C/N = 15:1, C/N = 20:1 and C/N = 25:1. Anhydrous glucose (99.97%) was added to the commercial diet to maintain elevated carbon ratio in the three treatments. Uninterrupted 24‐hr aeration was supplied during the test by using an air‐stone connected to an air pump to develop the bioflocs in the water column. The results indicated that BFT significantly reduced total ammonia nitrogen (TAN), nitrite (NO2^?‐N), total phosphorus (TP) concentrations and water transparency (Secchi depth) in C/N 15, C/N 20 and C/N 25 (p < .05), whereas the chemical oxygen demand (COD) and nitrate nitrogen (NO3^?‐N) concentrations gradually increased over time. In addition, weight gain, specific growth rate (SGR), survival and protein efficiency ratio (PER) were significantly elevated in C/N 20, while food conversion rate (FCR) was considerably decreased (p < .05) in C/N 20, compared with the control. Significant increases in lysozyme (LSZ), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) and alkaline phosphatase (AKP) activities of O. kaopingensis were found in the C/N 20 group after a 28‐day feeding trial (p < .05). Comparing the antioxidant capacity of O. kaopingensis in gills, brains, kidneys, hepatopancreas, intestines and serum of juveniles from the four experimental groups, the activity of superoxide dismutase (SOD), total antioxidant capacity (T‐AOC), catalase (CAT) activity and glutathione peroxidase (GSH‐PX) activity of juveniles in the C/N 20 group were significant higher (p < .05), and the content of malondialdehyde (MDA) was considerably lower, than in the control. Overall, these findings suggest that BFT not only can enhance O. kaopingensis growth performance and strengthen antioxidant status but also can upgrade immune response, ensuring the sustainable development of aquaculture.     

Effects of dietary soy isoflavones on growth,antioxidant status,immune response and resistance of juvenile grass carp (Ctenopharyngodon idella) to Aeromonas hydrophila challenge

The current study was conducted to evaluate the effect of dietary soy isoflavones (SI) on growth performance, antioxidant status, immune response and resistance to Aeromonas hydrophila in juvenile grass carp (Ctenopharyngodon idella). Six diets were formulated to contain 0 (control), 10, 50, 100, 500 or 1,000 mg SI per kg feed. Each diet was randomly allotted to triplicate net cages, and each net cage was stocked with 30 fish. The fish were fed one of the experimental diets to satiation twice per day for 60 days. The results showed that the WGR and DGC of the 500 mg/kg SI‐supplemented group were significantly higher than those of the non‐SI‐supplemented group (p < .05). Serum LZM and IgM activities in the SI‐supplemented groups were improved compared to the control group. SOD and GSH‐Px levels of fish fed the diet containing 500 mg/kg SI were significantly enhanced compared to those of fish fed the control diet (p < .05). Additionally, serum CAT, GSH‐Px and AKP activities in 50, 100 and 500 mg/kg SI‐supplemented groups were significantly higher than that in the control group (p < .05). The expression of most immune‐related genes (including IFN‐γ2, TNF‐α, M‐CSF2, IL‐6, IL‐12p40 and IL‐4) was significantly affected by dietary supplementation of SI. The group fed with 500 mg/kg SI had the highest 7‐day cumulative survival rate after challenge test (p < .05). The current results revealed that dietary inclusion of SI could improve the immune response and resistance against A. hydrophila and the supplementation level is suggested to be 500 mg/kg diet.     

Synergistic effect of sodium selenite and Pediococcus acidilactici on growth,intestinal bacterial counts,selenium bioavailability,hepatic enzymes and non‐specific immune response in rainbow trout (Oncorhynchus mykiss)

Lactic acid bacteria (LAB) have a crucial role in inorganic selenium metabolism as well as their known desirable effects on fish. In this study, the synergistic effects of dietary sodium selenite and Pediococcus acidilactici on growth performance, intestinal bacterial counts, selenium bioavailability, hepatic antioxidant enzyme thioredoxin reductase activity and hepatic glycolytic enzyme activity that is hexokinase, phosphofructokinase and pyruvate kinase, and non‐specific immune response such as serum lysozyme and complements C₃, C₄ and ACH₅₀ activity in rainbow trout (Oncorhynchus mykiss) were investigated. Thus, a 3 × 3 factorial design experiment was conducted with nine purified diets including three levels of sodium selenite (0, 1 and 2 mg/kg) and three levels of P. acidilactici (0, 7 and 9 log CFU/g). After 8 weeks of feeding, weight gain and specific growth rate were increased by increasing dietary sodium selenite and P. acidilactici levels compared to control (p < .05), whereas feed conversion ratio and condition factor was decreased by increasing dietary sodium selenite and P. acidilactici amounts in comparison with control (p < .05). Survival rate was not significantly affected among the experimental treatments (p > .05). Total cultivable bacterial populations after 4 and 8 weeks of the feeding trial were not significantly different among the dietary treatments, while LAB levels were higher in P. acidilactici‐fed groups than in control and selenium‐fed groups (p < .05). Selenocysteine, methylselenocysteine and selenomethionine levels in the intestine of rainbow trout were increased by increasing the sodium selenite and P. acidilactici levels (p < .05), and selenocysteine was found the most selenium species in the trout intestine. The quantity of total selenium in the whole body, intestine, blood, liver and muscle of rainbow trout were increased by increasing the amounts of sodium selenite and P. acidilactici compared to control (p < .05). Hepatic thioredoxin reductase and hexokinase activity were increased by increasing dietary selenium and P. acidilactici levels in comparison to control (p < .05), whereas phosphofructokinase and pyruvate kinase activity in the liver of rainbow trout were not significantly different between the dietary treatments. Serum lysozyme, complements C₃, C₄ and ACH₅₀ activity were enhanced by increasing dietary selenium and P. acidilactici levels compared to control (p < .05). The most synergistic effects of dietary supplements on growth and metabolism of rainbow trout were obtained at 2 mg/kg sodium selenite and 7 log CFU/g P. acidilactici. The findings revealed the synergistic effect of dietary selenium and P. acidilactici on growth and metabolism in rainbow trout (O. mykiss).     

Evaluation of some haemolymph biochemical properties and F‐cell prevalence in hepatopancreas of white leg shrimp (Litopenaeus vanammei) after fed diets containing apple cider vinegar and propionic acid

The present study aimed to evaluate the effect of dietary ACV (apple cider vinegar) and PA (propionic acid) supplementation on some biochemical properties of haemolymph and fibrillar cell (F‐cell) number in the hepatopancreas of shrimp. Five different diets supplemented with 0%, 1%, 2%, 4% ACV and 0.5% PA diets were fed to Litopenaeus vanammei. Some biochemical parameters of haemolymph (cortisol, alkaline phosphatase, albumin, urea, uric acid, creatinine, phosphorus, total hemocyte count) and prevalence of F‐cell in hepatopancreas were examined after 60 days feeding trial. The albumin concentration and total hemocyte count in 2% and 4% ACV and PA groups were significantly higher than those of other groups (p < .05). Moreover, the highest phosphorus concentration was detected in shrimp fed 4% ACV and PA experimental diets (p < .05). No significant differences in haemolymph cortisol, urea, uric acid, creatinine, alkaline phosphatase and tubule area were observed among the experimental groups (p > .05). 2% ACV group showed more F‐cell number in hepatopancreas than other groups while the reduction in F‐cell number was observed in the PA group. Based on these observations, the tested ACV and PA diets remarkably exhibited positive impacts on nutritional value and they may be a potential for adding to shrimp diet.     

Zingiber officinale and Glycyrrhiza glabra,individually or in combination,reduce heavy metal accumulation and improve growth performance and immune status in Nile tilapia,Oreochromis niloticus

This study evaluated the impact of dietary ginger and liquorice supplementation on growth performance, physiological and histopathological profiles and heavy metal accumulation in Nile tilapia fingerlings. Fish (n = 1,800, 17.5 ± 0.11 g BW) were randomly distributed into four treatment groups in triplicates and received no supplementation (control group), 5 ml aqueous ginger extract/kg feed (ginger group), 4 ml aqueous liquorice extract/kg feed (liquorice group) or 2.5 ml ginger plus 2 ml liquorice aqueous extracts/kg feed (mix group). The ginger‐liquorice mix supply improved the growth performance and feed efficiency (p < .05), increased the haematocrit and haemoglobin (p < .05), leucocytes (p = .108), neutrophils (p = .054), serum total protein (p < .05), albumin (p = .011) and globulin (p = .094) but decreased (p < .05) the blood urea nitrogen and creatinine than feeding liquorice or ginger lonely compared to the control. Heavy metal loads in pond water induced lamellar telangiectasis of gills and necrosis with sloughing of intestinal villi tips. These detrimental effects were alleviated, and the intestinal villus length (p = .041) and crypt depth (p = .069) were increased with liquorice supply. In all treatment groups, heavy metal contents in fish flesh were lower compared to the control. Thus, using ginger and/or liquorice aqueous extracts can decrease heavy metal accumulation in the fish flesh and exert positive effects on growth performance, metabolic profile and the intestinal and gill morphology of Nile tilapia.     

Effects of γ‐aminobutyric acid supplementation on the growth performance,serum biochemical indices and antioxidant status of pharaoh cuttlefish,Sepia pharaonis

This study evaluated the effects of dietary γ‐aminobutyric acid (GABA) on the growth performance, serum biochemical indices and antioxidant status of pharaoh cuttlefish, Sepia pharaonis. Cuttlefish were cultured in open‐culturing cement pool systems for 8 weeks. Six practical diets supplemented with graded levels of GABA (0, 20, 40, 60, 80 and 100 mg/kg) were formulated. Each diet was randomly assigned to triplicate groups of 60 cuttlefish (mean weight: 10.33 g), the cuttlefish were fed two times per day to apparent satiation. The results showed that the specific growth rate (SGR), weight gain (WG) and feed efficiency (FE) significantly increased with dietary GABA supplementation (p < .05). The survival rate (SR) and protein content in muscle significantly increased when 58.9 mg/kg GABA supplied. Moreover, the nitric oxide (NO) content and acid phosphatase (ACP) activity in serum were significantly increased with dietary GABA supplementation (p < .05), while the activity of aspartate aminotransferase (AST) in serum decreased significantly when supplied with GABA at 58.9 mg/kg (p < .05). In addition, dietary GABA improved antioxidation activity by significantly increasing the activities of superoxide dismutase (SOD) and catalase (CAT) but decreasing malondialdehyde (MDA) levels in the liver and gill (p < .05). On the basis of the quadratic regression analysis of FE, the optimum content of dietary GABA in S. pharaonis was estimated to be 55.3 mg/kg. The findings of this study demonstrated that dietary GABA had a positive effect on the growth performance, serum biochemical indices and antioxidant status of S. pharaonis.     

Effects of different levels of dietary protein on the growth performance,nutrient utilization,digestive enzymes and physiological status of white shrimp,Litopenaeus vannamei juveniles reared in inland saline water

A 60‐day feeding trial was conducted to illustrate the effect of dietary protein levels on the growth and physio‐metabolic responses of juvenile Litopenaeus vannamei reared in inland saline water (ISW). Six isoenergetic (15 MJ/kg) and isolipidic (60 g/kg) diets with graded level of crude protein viz., 240 (T240), 260 (T260), 280 (T280), 300 (T300), 320 (T320) and 340 (T340) g/kg diet were formulated. Significantly higher (p < .05) weight gain (%), specific growth rate, with lower food conversion ratio were found in T320 and T340 groups. The protein utilizing efficiency and whole‐body protein content were significantly higher (p < .05) in the T320 group. Trypsin activity increased with the increasing dietary CP level but amylase activity decreased with the increasing dietary CP level. Transaminase enzymes, haemolymph protein and haemocyanin were elevated in T320 and T340 groups. The lactate dehydrogenase and malate dehydrogenase activities were significantly higher (p < .05) in the T240 group. Shrimp of T240 group had the lowest (p < .05) glycogen and total haemocyte count with highest (p < .05) haemolymph glucose and antioxidant enzymes activities than the other groups. Based on the results, feeding 320 g CP/kg is found to be optimum for supporting maximum growth and health status of L. vannamei reared in ISW at 8 g/L salinity. The finding of the present study will help in developing a low‐cost feed for L. vannamei reared in ISW.     

Clostridium butyricum as probiotic for promoting growth performance,feed utilization,gut health and microbiota community of tilapia (Oreochromis niloticus × O. aureus)

The feeding trial was conducted to evaluate the potentials of Clostridium butyricum in the diet of tilapia. Fish (~14 g) were fed with basal diet supplemented with 0 (Control), 0.5 (C‐1), 1 (C‐2), 2 (C‐3), 4 (C‐4) and 8 (C‐5) g/kg commercial probiotic‐containing C. butyricum (1.5 × 10⁸ CFU/g) for 8 weeks. The results showed that weight gain significantly increased, and feed conversion ratio decreased in the C‐2, C‐3 and C‐4 groups (p < .05). The protein retention (except C‐1 group), lipid retention and apparent digestibility coefficient (ADC) of dry matter in probiotic supplementation groups were significantly enhanced, and ADC of protein in the C‐4 group was also improved (p < .05). The supplementation of probiotic significantly increased villus height in anterior intestines and reduced the numbers of intestinal Escherichia coli (p < .05). High‐throughput sequencing showed that top three phyla namely Planctomycetes in all probiotic‐containing groups, Proteobacteria in the C‐1 and C‐2 groups and Chloroflexi in the C‐3 group had higher level than the NC group. The cumulative mortality was reduced by dietary probiotic after challenging with Aeromonas hydrophila (p < .05). In conclusion, C. butyricum can be supplemented at 1–2 g/kg feed for promoting the growth, feed utilization, gut health and microbiota of tilapia.     

Influences of dietary antimicrobial peptide APSH‐07 on the growth performance,immune response and vibriosis resistance of abalone Haliotis discus hannai Ino

A 115‐day feeding trial and subsequently a 10‐day challenge test with Vibrio parahaemolyticus were conducted to investigate the effects of dietary antimicrobial peptide APSH‐07 on growth performance, immune response, antioxidative status and vibriosis resistance of abalone Haliotis discus hannai (initial body weight: 2.06 ± 0.01 g; initial shell length: 25.42 ± 0.18 mm). Four artificial diets were designed with 0 (artificial diet control), 7.5, 15.0 and 22.5 mg/kg of APSH‐07, respectively. The brown alga Laminaria japonica was used as the live food control. Results showed that the specific growth rates of abalone in the groups with 7.5 and 15.0 mg/kg dietary APSH‐07 were significantly higher than those in the controls (p < .05). The total haemocyte counts and respiratory burst activity in haemolymph of abalone in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the groups with 0 and 22.5 mg/kg of dietary APSH‐07 (p < .05). The gene expression levels of Mn‐superoxide dismutase and thioredoxin peroxidase 2, and the activities of glutathione peroxidase in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the other groups (p < .05). Cumulative mortality of abalone after the challenge test was significantly decreased in the group with 7.5 mg/kg of dietary APSH‐07 supplementation. Supplementation of 22.5 mg/kg dietary APSH‐07 significantly increased the cumulative mortality. In conclusion, 7.5 mg/kg of dietary APSH‐07 supplementation had the better growth performance, higher antioxidation, immune and disease resistance capacity of abalone. Excessive supplementation of dietary antimicrobial peptide APSH‐07 (22.5 mg/kg) had significantly negative effects. Further studies are needed to determine the optimal level of dietary APSH‐07 supplementation for abalone.     

The supplementation of dietary selenium yeast and green tea‐derived polyphenols improves antioxidant capacity and immune response in juvenile Wuchang bream under ammonia stress

In order to investigate protective effects of dietary selenium yeast and green tea‐derived polyphenols on ammonia stress, healthy Wuchang bream juveniles (3.24 ± 0.08 g) were fed basal diets and three treatment diets supplemented with 0.50 mg/kg selenium yeast, 50 mg/kg green tea‐derived polyphenols, and combination of 0.50 mg/kg selenium yeast and 50 mg/kg green tea‐derived polyphenols, respectively. After a 60‐day feeding, fish were exposed to 22.5 mg/L total ammonia nitrogen for 48 hr. The results showed that total antioxidant capacity, catalase and glutathione peroxidase in the serum were higher in three treatment groups than in control group before and after ammonia exposure (p < .05), while serum cortisol was lower in three treatment groups (p < .05). Similarly, the content and mRNA levels of splenic interleukin 1β, tumour necrosis factor α and immunoglobulin M showed significant increases in three treatment groups in relative to the controls (p < .05). Splenic lesions for three treatment groups were less severe, and the healing process of the lesions was faster than the control group after ammonia exposure. These findings demonstrated that single and combined supplementation of selenium yeast and green tea‐derived polyphenols could enhance antioxidant and immune responses to alleviate oxidative stress and damage by ammonia. Overall, the combination of selenium yeast and green tea‐derived polyphenols has a potential use as a preventive or therapeutic measure in ammonia‐exposed fish.     

Effect of supplementation of dietary fibre concentrates on biochemical parameters,stress response,immune response and skin mucus of jundiá (Rhamdia quelen)

The aim of this study was to assess the effects of different dietary fibre concentrates (DFC: Mucilage = MG; pectin = PN or β‐glucan+mannan = βg+M), on biochemical parameters, stress and immune response and skin mucus of jundiá (Rhamdia quelen). The fish (7.16 ± 0.06 g) were fed with Control diet (0 g/kg of DFC); diet supplemented with 5 g/kg of commercial prebiotic (CP 5) or diets supplemented with 5 or 10 g/kg of MG; PN or βg+M. After 8 weeks of the feeding trials, biochemical parameters (cholesterol, glucose, albumin and total protein), cortisol, immunoglobulin IgM and mucoproteins of skin mucus were assessed. Results demonstrated that supplementation with PN increased cholesterol levels (p<.05). After application of the stressor, most fish, except those in the PN and 10 g/kg MG groups, showed significant increases (p<.05) in cholesterol, glucose and albumin levels. The jundiás showed no difference in cortisol levels after application of the stressor (p>.05). IgM levels were significantly high in fish supplemented with DFC (p<.05). However, the concentration of mucoproteins in skin mucus was not influenced in the different treatments (p>.05). The results showed that supplementation with DFC promoted beneficial effects on the metabolism of jundiá.     

Effects of dietary zinc level on performance,zinc status,tissue composition and enzyme activities of juvenile Siberian sturgeon,Acipenser baerii (Brandt 1869)

This study conducted to evaluate the effect of dietary zinc (Zn) levels on feed utilization, tissue Zn composition and serum enzyme activities of juvenile Siberian sturgeon, Acipenser baerii. Five isoenergetic and isonitrogenous semi‐purified diets were formulated with increasing Zn sulphate (ZnSO₄.5H₂O) level to provide the actual Zn values of 14.7 (control), 20.8, 27.3, 37.7 and 46.4 mg/kg diet. Each diet was assigned to three groups of 20 experimental Siberian sturgeons with uniform size (initial weight of 26.52 ± 0.94 g) for a period of 8‐week feeding trial. Results showed that growth performance and muscle protein content were significantly increased with increasing dietary Zn level up to 27.3 mg/kg (p < .05), beyond which they remained significantly unchanged (p > .05). Muscle lipid content significantly declined with increasing dietary Zn level. While muscle and serum Zn contents were not significantly changed among treatments (p > .05), liver Zn content tended to rise with increasing dietary Zn supplementation. Alkaline phosphatase, superoxide dismutase and glutathione peroxidase activities were also raised with increasing dietary Zn level. The adequate amount of dietary Zn requirements for the Siberian sturgeon was estimated to be 28.24 mg/kg based on the relative growth rate and 34.60 mg/kg based on the liver Zn content.     

Dietary supplementation of Tribulus terrestris extract improves growth and reproductive performances of the male Nile tilapia (Oreochromis niloticus)

Effects of dietary supplementation of Tribulus terrestris (TT) and 17α‐methyl testosterone (MT) on growth performance and reproductive efficiency of male Nile tilapia, Oreochromis niloticus (N = 75, initial weight 60 ± 3.251 g), were evaluated. Five experimental treatments in triplicate were fed on basal diet (30% crude protein) (control group, CG), basal diet containing 60 mg MT (MT group) and basal diet containing 250, 500 and 750 mg/kg diet TT (TT250, TT500 and TT750 groups), respectively, for consecutive 45 days. Results revealed that the fish received TT in diet showed better growth performance (final body weight, weight gain, feed conversion ratio, specific growth rate, testes weight and gonadosomatic index) compared with MT and CG (p < .05), as well as for haemoglobin content and red blood cell count (p < .05). The highest level of testosterone, 11‐keto testosterone, sperm concentration and vitality was recorded in TT750 followed by TT500, TT250 and MT groups, while the lowest value was recorded in the CG (p < .05). The highest seminiferous tubule diameter was recorded in TT750 followed by TT500, TT250 and CG, while the lowest diameter was recorded in the MT group (p < .05). Fish received TT showed normal dilated seminiferous tubule filled with large amount of free sperms within their lumen in a dose related effect. In conclusion, male Nile tilapia dietary supplementation with different inclusions of Tribulus terrestris extract specially 500 and 750 mg/kg diet improves growth performance, health condition, semen quality and reproductive efficiency without any harmful effect on water quality.     

Effects of dietary marine microalgae,Tetraselmis suecica,on production,gene expression,protein markers and bacterial count of Pacific white shrimp Litopenaeus vannamei

The marine flagellated Chlorophyta Tetraselmis suecica is among the most important live food species in marine aquaculture. In the present study, the effects of dietary supplementation of dried marine microalgae, Tetraselmis suecica, on growth performance; feed utilization; chemical composition; gene expression of superoxide dismutase (SOD), glutathione peroxidase (GPx) and insulin‐like growth factor 2 (IGF‐II) gene of Pacific white shrimp, Litopenaeus vannamei; muscle protein polymorphism; and microbial count were assessed and evaluated. Three hundred and sixty L. vannamei (postlarvae) Pls (0.124 ± 0.002 g) were randomly stocked into 40‐L glass aquaria (30 shrimp/aquarium) and fed three times daily four tested diets: a basal diet (control), diet incorporated with 2.5 g kg^?1 dried T. suecica (T1), 5 g kg^?1 dried T. suecica (T2) and 7.5 g kg^?1 dried T. suecica (T3) in triplicates, for 90 days. At the end of the trial, the survival rate (SR) of L. vannamei fed diets supplemented with different levels of T. suecica was significantly (p < .05) higher than the control diet. The highest weight gain and specific growth rate and the best feed conversion ratio were recorded on L. vannamei fed a diet supplemented with a 7.5 g/kg dried T. suecica. The highest protein, lipid and ash contents were obtained in L. vannamei fed the diet containing 7.5 g/kg T. suecica, when compared with the remaining tested diets. The gene expression of antioxidant genes SOD and GPx was the lowest in the T3 group in comparison with the control group. Meanwhile, expression level of IGF‐II was higher in the T2 group. The total heterotrophic bacterial count was significantly (p < .05) increased with the cumulative T. suecica level, while no significant (p > .05) differences were found in the total Vibrio count among treatments. Overall, the present results have shown that the diet supplemented with the highest inclusion level of dried T. suecica resulted in improved growth and nutrient utilization.     

The antioxidant status,apoptosis, intercellular integrity and immune function of grass carp (Ctenopharyngodon idella) head kidney and spleen fed different levels of pyridoxine

This study was carried out to investigate the effects of dietary pyridoxine (PN) on the antioxidant status, apoptosis, intercellular integrity and immune function of head kidney and spleen in young grass carp (Ctenopharyngodon idella). Results showed that compared with the optimal PN level, (a) PN deficiency decreased antioxidant enzyme activities and down‐regulated the mRNA levels of antioxidant enzymes, NF‐E2‐related factor 2 (Nrf2), myeloid cell leukaemia‐1 (Mcl‐1) and tight junction proteins, whereas it increased the contents of reactive oxygen species (ROS), malondialdehyde (MDA) and protein carbonyl (PC), and up‐regulated the mRNA levels of cysteinyl aspartic acid‐protease (caspase), Fas ligand (FasL), p38 mitogen‐activated protein kinase (p38MAPK) and myosin light chain kinase (MLCK) in the head kidney and spleen of fish (p < .05). (b) PN deficiency decreased lysozyme (LZ) and acid phosphatase (ACP) activities, and complement contents, and down‐regulated the mRNA levels of antibacterial peptides, anti‐inflammatory cytokines, inhibitor of κBα (IκBα) and target of rapamycin (TOR), whereas it up‐regulated the mRNA levels of pro‐inflammatory cytokines and nuclear factor kappa B p65 (NF‐κBp65) in the head kidney and spleen of fish (p < .05). (c) Dietary PN requirements of grass carp based on the malondialdehyde content and lysozyme activity were estimated as 4.97 and 4.99 mg/kg diet, respectively.     

Supplementation of heat‐inactivated Bacillus clausii DE5 in diets for grouper,Epinephelus coioides,improves feed utilization,intestinal and systemic immune responses and not growth performance

In recent years, more and more attentions have been paid to the development and application of probiotics in aquaculture, and viable probiotics have been extensively studied, while rare information was available about inactivated probiotics in aquaculture. Therefore, in this study, a feeding trial was designed to investigate the effect of dietary supplementation of heat‐inactivated probiotic Bacillus clausii DE5 on growth performance, immune response and key immune genes expression in head kidney and intestine in grouper Epinephelus coioides. Fish were fed for 60 days with control diet (C) and two experimental diets containing 1.0 × 10⁸ CFU/g live (T1) and heat‐inactivated (T2) B. clausii DE5, respectively. The probiotic treatments did not affect the final weight, weight gain (WG) and specific growth rate (SGR) of E. coioides at days 30 and 60 (p > .05), while both heat‐inactivated and live B. clausii DE5 significantly decreased the feed intake and feed conversion ratio (FCR) at day 60 (p < .05). Serum lysozyme activity and complement C3 level in the two probiotic treatments were significantly higher than those in the control (p < .05). The lysozyme activity and complement C3 level at day 60 were significantly higher than those at day 30 (p < .05), while no significant interaction effect between diet and administration date was observed. Moreover, the heat‐inactivated B. clausii DE5 significantly improved the expression of TLR5, pro‐inflammatory cytokines (IL‐8 and IL‐1β) and TGF‐β1 in head kidney and intestine (p < .05), while the live probiotic did not show any significant effect on the expression of these key immune‐related genes in head kidney and intestine. These results indicate that dietary supplementation of heat‐inactivated B. clausii DE5 effectively improved feed utilization and both the local and systemic immune responses of E. coioides.     

Effect of dietary soybean lecithin on growth parameters,digestive enzyme activity,antioxidative status and mucosal immune responses of common carp (Cyprinus carpio)

Common carp, Cyprinus carpio (13.54 ± 0.48 g), were fed soybean lecithin‐incorporated diets for 8 weeks. After the feeding trials, the different groups of fishes were assessed by survival, growth parameters, digestive enzymes, antioxidant enzyme activities, mucosal immune parameters, etc. The lecithin‐incorporated diets (10–30 g kg^–1) helped to increase the weight gain significantly (p < .05) compared with control groups; activity of digestive enzymes was also significantly (p < .05) higher compared with experimental groups. The antioxidant enzymes including catalase and serum superoxide dismutase were significantly (p < .05) higher in the C. carpio fed lecithin‐incorporated feed than the ones fed the control feed. The serum thiobarbituric acid‐reactive substances and serum glutathione peroxidase activity did not differ significantly (p > .05) between control and experimental groups. The positive correlation supported that antibacterial activity was significantly (p < .05) increased with increasing concentration of lecithin in the diets. Among the different mucosal immune parameters, lysozyme, protease and esterase activities were significantly (p < .05) increased with lecithin‐incorporated diets. The results of this study confirm that 20 and 30 g kg^–1 lecithin inclusion in diets helped to enhance the activities of digestive enzymes, antioxidant enzymes, antibacterial factors and mucosal immune parameters in C. carpio.     

The optimal dietary protein level of juvenile silver sillago Sillago sihama at three dietary lipid levels

A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg.