Effect of dietary eucalyptol on stress markers,enzyme activities and immune indicators in serum and haematological characteristics of common carp (Cyprinus carpio) exposed to toxic concentration of ambient copper

In the present study, protective effects of dietary eucalyptol were investigated against copper toxicity in common carp (Cyprinus carpio). The fish were fed with diets supplemented with 0 (control), 0.5% and 1% eucalyptol for 14 days before exposure to 0.25 mg/L ambient copper for 7 days. Blood samples were taken from the fish before and 1, 3 and 7 days after copper challenge. Before copper challenge, eucalyptol significantly decrease serum aspartate aminotransferase (AST) activity, but increase blood WBC, RBC, hematocrit and haemoglobin, serum lysozyme, total immunoglobulin (Ig) compared with the control. Copper exposure led to stress, tissue damage and immunosuppression. After the copper challenge, significant elevation in serum cortisol, glucose, alanine aminotransferase (ALT) and AST, but decrease in blood RBC and WBC, and serum lysozyme, complement (ACH50) and total immunoglobulin (Ig) was observed. Eucalyptol administration significantly mitigated the increase in serum cortisol, glucose, ALT and AST, and decrease in WBC, ACH50, lysozyme and total Ig. Dietary 1% eucalyptol had significantly higher protective effects than 0.5% in the case of serum AST, lysozyme, total Ig and ACH50. In conclusion, eucalyptol has beneficial health effects on common carp and enables to mitigate copper‐induced stress, tissue damage, and immunosuppression. Eucalyptol at the dietary 1% level is recommended to protect common carp against adverse effects of copper toxicity.     

Anaesthetic efficacy and biochemical effects of 1,8‐cineole in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792)

The aim of the present study was to investigate the anaesthetic efficacy and biochemical effects of 1,8‐cineole (cineole) in rainbow trout (Oncorhynchus mykiss). The fish were exposed to 200, 300, 400, 500, 600 and 800 μl/L cineole and time of induction of anaesthesia and recovery from anaesthesia were recorded. Thereafter, the fish haematological and biochemical responses to anaesthesia with different concentrations of cineole were studies. Moreover, the haematological and biochemical response of fish anaesthetized for 300 s with either cineole (283 μl/L) or eugenol (25 μl/L) were compared. Cineole at the concentrations of 200–800 μl/L induced stages 2, 3 and 4 anaesthesia within 109‐29.3, 226‐59 and 418‐117 s respectively. Increase in anaesthesia induction time led to higher stress responses and enzymes’ activity characterized by elevation in red blood cell (RBC), white blood cell (WBC), blood haematocrit and haemoglobin, and plasma cortisol, glucose and lactate levels, and aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase and creatine phosphokinase (CPK) activities. However, cineole concentrations had no significant effects on plasma alanine aminotransferase (ALT) activity and ions levels. Comparison of cineole with eugenol showed that cineole showed less change in blood RBC and plasma AST, ALP, CPK, cortisol and glucose levels compared to eugenol. In conclusion, cineole is efficient to anaesthetize trout at concentrations of 200–800 μl/L. Increase in cineole concentration shortens anaesthesia induction time, stress response and probably tissue damage. The concentrations of 600–800 μl/L cineol is recommended for rapid sampling as it causes the least stress and enzymatic responses. The present results suggest that cineole causes slightly lower side effects in trout compared to eugenol.     

A comparative study on the effects of a pesticide (cypermethrin) and two metals (copper, lead) to serum biochemistry of Nile tilapia, Oreochromis niloticus

The present study was designed to compare the responses in freshwater fish Oreochromis niloticus exposed to a synthetic pyrethroid, cypermethrin (CYP); an essential metal, copper (Cu); and a nonessential metal, lead (Pb). Fish were exposed to 0.05 μg/l CYP, 0.05 mg/l Cu, and 0.05 mg/l Pb for 4 and 21 days, and the alterations in serum enzyme activities, metabolite, and ion levels were determined. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities increased in response to CYP, Cu, and Pb exposures at both exposure periods. While elevations in alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activities and in cholesterol level were observed in pesticide-exposed fish at 4 and 21 days, they increased in Cu- and Pb-exposed fish at 21 days. Although metal-exposed fish showed increases in cortisol and glucose levels at 4 days followed by a return to control levels at the end of the exposure period, their levels elevated in pesticide-exposed fish at both exposure periods. Total protein levels decreased in Pb- and pesticide-exposed fish at 21 days. Na⁺ and Cl⁻ levels decreased in pesticide-exposed fish at both exposure periods and in Cu- and Pb-exposed fish at 21 days. The exposures of pesticide and metals caused an elevation in K⁺ level at the end of the exposure period. The present study showed that observed alterations in all serum biochemical parameters of fish-treated pesticide were higher than those in fish exposed to metals.     

Effects of Neutral Phytase Supplementation on Biochemical Parameters in Grass Carp,Ctenopharyngodon idellus,and Gibel Carp,Carassius auratus gibelio,Fed Different Levels of Monocalcium Phosphate

Two 8‐wk studies were conducted to evaluate the effects of neutral phytase supplementation on hemato‐biochemical status, liver biochemical parameter, and intestinal digestive enzyme activity of grass carp, Ctenopharyngodon idellus, and gibel carp, Carassius auratus gibelio, fed with different levels of monocalcium phosphate (MCP). The control diet was prepared with 2% MCP but without phytase (P2.0). The other three experimental diets were prepared with the addition of 1.5, 1.0, and 0.5% MCP, respectively, when supplemented with 500 U/kg neutral phytase in each diet and designated as PP1.5, PP1.0, and PP0.5, respectively. The results indicated that the serum alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate transaminase (AST) activities, as well as the albumin (ALB) content were increased in grass carp (P < 0.05) and gibel carp (P > 0.05) fed with phytase‐supplemented diets. Meanwhile, the serum cholesterol, high‐density lipoprotein, and total protein contents of the two species of fish were increased in comparison to the control. In addition, dietary phytase inclusion did not significantly affect hepatic ALP, ALT, and AST activities in the two species of carp fed with different levels of MCP. Amylase activity increased in foregut and hindgut of both species when fed with the phytase‐supplemented diets while lipase activity was reduced in the foregut and hindgut in both fish. This study suggests that neutral phytase supplementation increases serum ALP, ALT, and AST activities but does not notably affect these enzyme activities in the liver of the two species of carp when fed different levels of MCP. On the other hand, amylase activity increased while lipase activity was reduced in the intestine of the species of carp fed with phytase‐supplemented diets.     

Dietary supplementation with taurine improves ability to resist ammonia stress in hybrid snakehead (Channa maculatus♀ × Channa argus♂)

This study determined the effect of dietary supplementation with taurine on plasma biochemical indices, blood cell apoptosis rate, survival rate and expression of catalase (CAT), glutathione reductase (GR) and heat‐shock protein 70 (HSP70) gene in juvenile hybrid snakehead under ammonia stress. Six diets were formulated to contain 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g/kg taurine. Each diet was fed to triplicate groups of fish in cylindrical tanks. After 8 weeks of feeding, 20 fish per tank were exposed to ammonia stress (total ammonia nitrogen = 200 mg/L) for 48 hr. The results showed that, after ammonia stress, plasma alkaline phosphatase (ALP), triacylglycerol (TG) levels and survival rate significantly decreased (p < 0.05) while plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN), respiratory burst, blood cell apoptosis rate, hepatic CAT, GR and HSP70 mRNA expression levels significantly increased (p < 0.05). On the other hand, dietary supplementation with taurine significantly reduced levels of BUN, respiratory burst and blood cell apoptosis rate. Supplemented groups significantly increased relative mRNA expression of hepatic CAT, GR and HSP70 as well as increased survival rate (p < 0.05). These results indicated that dietary supplementation with taurine improved ability to resist ammonia stress in hybrid snakehead.     

Determination of normal values of some blood serum enzymes in <Emphasis Type="Italic">Acipenser stellatus</Emphasis> Pallas

Hematological studies on fishes have assumed greater significance due to the increasing emphasis on pisciculture and greater awareness of the pollution of natural water resources. Such studies have generally been used as an effective and sensitive index to monitor physiological and pathological changes in fishes, especially in the management of endangered species. This study was undertaken to establish a reference range for six important blood serum enzymes. Serum samples of 40 Acipenser stellatus (20 female and 20 male) were analyzed, and serum enzyme values were determined. The reference ranges for the total samples and also for each sex were determined. The activities of measured enzymes in serum were: aspartate aminotransferase (AST) 265.60 ± 56.55 IU/l, alanine aminotransferase (ALT) 5.65 ± 1.18 IU/l, acid phosphatase (ACP) 15.63 ± 2.59 IU/l, lactate dehydrogenase (LDH) 2007.15 ± 521.97 IU/l, creatine kinase (CK) 6,596.05 ± 1,807.19 IU/l, and alkaline phosphatase (ALP) 69.05 ± 13.04 IU/l. There were significant differences between male and female fish in terms of AST and CK activity (P < 0.05). These may be related to the season of sampling and changing physiological cycles during spawning during which period the sexual hormones and stress are more obvious. However, there were no differences in the activity of ALP, ACP, LDH, and ALT between sexes. The correlations between measured enzymes were also determined.     

Effect of date palm (Phoenix dactylifera) seed extract as a dietary supplementation on growth performance immunological haematological biochemical parameters of common carp

This study aimed to assess the effect of date palm (Phoenix dactylifera) seed (DPS) extract at 0.5%, 1%, 2% and 4% as a dietary supplement over 60 days, on the growth and immune‐physiology of common carp (Cyprinus carpio) fingerlings. The weight gain in fish fed with 0.5% DPS was significantly greater than other treatments and fish fed a control diet with no DPS (p < 0.05). With increasing DPS dosage above 0.5%, weight gain and growth rate gradually declined. Feed conversion ratio in fish fed 0.5% DPS was lower than other treatments, but not significantly so, except for the highest DPS concentration (4%). With respect to immunophysiological variables, fish treated with 0.5% DPS leucocyte, neutrophil, lymphocyte and monocyte counts that were generally equivalent to fish on a control diet, but significantly lower than fish treated with greater concentrations of DPS (p < 0.05). Chemilumincene response and lysozyme activity, as well as levels of total protein and albumin, were higher in all treated groups than the control. The highest levels of iron and high density lipoprotein (HDL) were observed in fish fed with 0.5% DPS, while the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP), glucose, cholesterol, triglyceride, low density lipoprotein (LDH), thyrotropin, thyroxine and triiodothyronine were lower level in fish fed with 0.5% DPS than in all other treatments. In aquaculture practice, use of 0.5% DPS in commercial carp feed can improve the growth rate and condition of fish.     

Changes in metabolic enzymes,cortisol and glucose concentrations of Beluga (<Emphasis Type="Italic">Huso huso</Emphasis>) exposed to dietary methylmercury

In this paper, effects of dietary methylmercury (MeHg) on several blood biochemical parameters including GLU (glucose), LDH (lactate dehydrogenase), AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase) and cortisol were investigated in the Beluga sturgeon (Huso huso). Beluga juveniles were fed for 32 days on four diets containing MeHg (control: 0.04 mg kg⁻¹; low: 0.76 mg kg⁻¹; medium: 7.88 mg kg⁻¹; and high 16.22 mg kg⁻¹ treatment). Significant increases (P < 0.05) were observed in all biochemical parameters, except ALP levels, which decreased significantly (P < 0.05) compared to the control group with either dose- or time-dependent effects. These results suggest that long-term dietary MeHg exposure may affect metabolic enzyme activity and glucose levels in Belugas. These findings provide useful information for environmental and fishery officials to apply in future decisions for managing fish resources in Caspian Sea.     

Effects of long-term silymarin oral supplementation on the blood biochemical profile of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Silymarin, an extract from “milk thistle” (Silybum marianum) plant is traditionally used as herbal medicine. The present study was conducted to investigate the clinical effects and possible side effects of silymarin on biochemical blood parameters of rainbow trout (Oncorhynchus mykiss). Fishes were treated with 0 (control), 100, 400, and 800 mg of silymarin per kg of food during 4 weeks. Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine kinase (CK), glucose, total protein, creatinine, triglyceride, cholesterol, urea, uric acid and liver cellular total antioxidant, and protein content were measured after 7, 14, and 28 days of silymarin treatment. The results showed that oral administration of silymarin in fish significantly reduced plasma glucose and cholesterol levels and relatively increased plasma total protein and globulin concentrations (P < 0.05). Increasing plasma albumin levels indicate the important role of albumin in drug transportation in circulatory system of fish. Silymarin also stabilized cellular membrane structure and regulated the levels of AST, ALT, ALP, CK, and LDH activity. In conclusion, on the basis of these results, oral administration of silymarin up to 400 mg per 1 kg of food has no side effect on blood biochemical and clinical parameters of fishes. However, oral administration of 800 mg/kg- of silymarin caused cytotoxicity and modifications in blood biochemical parameters of fish.     

Effects of water temperatures and dietary protein levels on growth,body composition and blood biochemistry of juvenile GIFT tilapia (Oreochromis niloticus)

This study was carried out to evaluate the effects of water temperature (WT) and dietary protein levels on growth, body composition and blood biochemistry of GIFT tilapia (Oreochromis niloticus; initial average body weight: 38.75 ± 0.61g, n = 20). The fish were fed with six diets contained graded levels of protein (209.6, 251.0, 302.4, 354.3, 401.9 and 456.8 g/kg) and raised in each of WT (22°C, 28°C and 34°C) for 8 weeks. Results showed that the growth, feed utilization and protein efficiency were significantly increased with the increase of dietary protein level and followed by the latter platform at each WT. The fish raised at 28 and 34°C showed similar growth performance, and their values were significantly higher than the fish raised at 22°C (p < 0.05). The lipid contents in whole body were decreases with the increase of dietary protein level at each WT. However, the whole‐body protein, ash and moisture contents were not affected by dietary protein level or WT (p > 0.05). The values of serum biochemical indices (ALP, ALT, AST and TCHO) were all decreased with the increase of dietary protein. The optimal dietary protein requirement for GIFT tilapia to achieve maximal growth performance is 374.4, 301.7 and 304.9 g protein/kg diet at 22, 28 and 34°C, respectively.     

Beneficial effects of thyme (Thymus vulgaris) extract on oxytetracycline‐induced stress response,immunosuppression, oxidative stress and enzymatic changes in rainbow trout (Oncorhynchus mykiss)

The aim of the present study was to investigate the effects of thyme extract (TE) and oxytetracycline (OTC) on rainbow trout (Oncorhynchus mykiss) stress response, immune parameters, oxidative stress and enzymatic changes. The fish were assigned into eight treatments receiving diets with OTC (0 and 2.5 g/kg) and TE (0, 5, 10 and 20 g/kg) for 2 weeks. Thereafter, serum characteristics and gut oxidative status were evaluated. OTC significantly increased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and cortisol and glucose levels; however, TE significantly suppressed these changes. OTC significantly decreased, but TE significantly increased gut catalase (CAT) and glutathione peroxidase (GPx) activities. OTC significantly decreased gut superoxide dismutase (SOD) and increased gut glutathione‐s‐transferase (GST) activities, malondialdehyde (MDA) content and blood monocyte percentage; TE significantly mitigated these changes. TE significantly increased serum total protein, total Ig, lysozyme and ACH50, and blood WBC; however, OTC significantly decreased serum lysozyme and ACH50 activities and blood WBC and neutrophil percentage. In conclusion, OTC administration causes immunosuppression, which might be related to deterioration of fish health. TE augments trout innate immune and antioxidant status and is capable to counteract OTC‐induced health and immune deterioration.     

Effects of dietary nucleotides supplementation on rainbow trout (Oncorhynchus mykiss) performance and acute stress response

This experiment was conducted to examine the effect of dietary nucleotides (NT) on fish performance and acute stress response on fingerling rainbow trout (23 g ± 0.01, mean weight ± SEM). Five experimental diets according to different levels of supplemented nucleotides (0, 0.05, 0.1, 0.15, and 0.2%) were assayed on experimental fish for 8 weeks. Growth, hematological parameters (hematocrit, hemoglobin, erythrocyte, lymphocyte, and neutrophil count), serum proteins (globulin, albumin), and plasma enzymatic activity (alkaline phosphatase, ALP; aspartate transaminase, AST; lactate dehydrogenase, LDH; alanine transaminase, ALT) were assayed. At the end of feeding trial, fish fed the control and 0.2% diets were subjected to handling and crowding stress. Modulatory effects of nucleotides on acute stress response (cortisol and glucose) and plasma electrolytes (Na⁺, Cl⁻, K⁺, and Ca²⁺) were studied. The percentage of body weight gain (WG) and feed efficiency (FE) of fish were better when the fish were fed 0.15–0.2% diets. Fish fed the nucleotide-supplemented diets tended to have lower levels of serum enzymes including ALP, AST, LDH, and ALT. Plasma cortisol levels of fish fed on 0.2% diet under handling and crowding stress were significantly lower than fish fed the control diet at all post-stress time intervals. In our study, fish fed nucleotide-supplemented diet had significantly lower concentrations of glucose compared to those fed the basal diet. The concentrations of sodium, chloride, calcium, and potassium of fish fed the control diet were significantly lower than in fish fed nucleotide-supplemented diet. Dietary nucleotides administration seems to promote growth and to enhance resistance against handling and crowding stress in fingerling rainbow trout.     

Non‐specific immune responses and intestinal immunity of common carp (Cyprinus carpio) fed Jujube (Ziziphus jujube) fruit extract

This study investigated the effects of Jujube (Ziziphus jujube) fruit extract (JFE) as an excellent source of nutrients and phytochemicals on the innate immune responses and expression of genes involved in intestinal immunity of common carp (Cyprinus carpio) fingerlings. Three hundred healthy carps (10.78 ± 0.05 g) fingerlings were randomly allocated to 12 fibre glass tanks (300L; 15 fish per tank) and fed for 8 weeks with experimental diets which contained different levels of JFE (0%, 0.25%, 0.5%, or 1%). Thereafter, serum non‐specific immune parameters [total immunoglobulin (Ig) as well as alternative complement (ACH50) and lysozyme activity (SL)] and expression of genes involved in intestinal immunity (lyz, tnf‐alpha, il1b, il8, il10, and tgfb) were measured. The results revealed a significant elevation of Ig level in the JFE‐fed fish (p < 0.05) over the control fish. However, there was no significant difference among treated groups in case of serum total Ig. The highest levels of SL and ACH50 were observed in fish fed 1% of JFE. Nonetheless, no significant difference was noticed regarding SL and ACH50 of control and other supplemented groups (0.25% and 0.5% JFE). Also, expression of lyz, tnf‐alpha, and il1b genes increased significantly (p < 0.05) in JFE‐fed fish, in a dose‐dependent manner. On the other hand, significant (p < 0.05) down‐regulation of il110 and tgfb were noticed in treated groups. The present findings suggested that extraction of Jujube, Z. jujube fruit, possesses beneficial effects on immune responses of common carp.     

Plasma biochemical responses of the omnivorous crucian carp (<Emphasis Type="Italic">Carassius auratus</Emphasis>) to crude cyanobacterial extracts

Healthy crucian carp (Carassius auratus) were treated by intraperitoneal (i.p.) injection of crude cyanobacterial extracts at two doses, 50 and 200 μg MC-LR equiv kg⁻¹ BW. High mortality (100%) was observed within 60 h post injection in the high-dose group. In the treated fish, activities of four plasma enzymes, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), all showed substantial increases, with both dose and time-dependent effects. These increases of enzyme activity indicate severe impairment occurred in the liver of crucian carp over time. Plasma concentrations of energy-related biomolecules including glucose (GLU), cholesterol (CHO), triglyceride (TG), and total protein (TP) showed marked changes in the high-dose group, possibly a nutritional imbalance correlated with the liver injury caused by intraperitoneal exposure to crude cyanobacterial extracts.     

Dietary microbial levan ameliorates stress and augments immunity in Cyprinus carpio fry (Linnaeus, 1758) exposed to sublethal toxicity of fipronil

A 45‐day feeding trial was conducted to study the stress ameliorating and immunomodulatory role of microbial levan in Cyprinus carpio fry exposed to sublethal dose (1/10th LC₅₀) of fipronil [(±)‐5‐amino‐1‐(2,6‐dichloro‐α,α,α‐trifluoro‐p‐tolyl)‐4‐trifluoromethylsulfinylpyrazole‐3‐carbonitrile]. Two hundred and twenty‐five fry were randomly distributed in five treatments in triplicates. Four purified diets were prepared with graded levels of microbial levan. Five different treatment groups were levan control L₀P₀ (basalfeed + 0% levan without exposure to pesticide); pesticide control L₀P₁ (basalfeed + 0% levan with exposure to pesticide); L_0.25P₁ (basalfeed + 0.25% levan with exposure to pesticide); L_0.50P₁ (basalfeed + 0.50% levan with exposure to pesticide) and L_0.75P₁ (basalfeed + 0.75% levan with exposure to pesticide). Lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and fructose‐1,6‐diphosphatase (FDPase) activites were significantly (P < 0.05) increased, whereas alkaline phosphatase (ALP), adenosine triphosphatase (ATPase) and acetyl choline esterase (AchE) activities were significantly (P < 0.05) reduced in higher levan‐fed groups. RBC, haemoglobin and WBC counts were significantly (P < 0.05) increased in the levan‐fed groups. Similar trends were also observed for the total serum protein, globulin, NBT and lysozyme activities. Blood glucose and serum cortisol exhibited a third order polynomial relationship with increasing level of dietary levan. Overall result showed stress ameliorating, immunostimulating and protective role of microbial levan against fipronil‐induced stress in C. carpio fry at 0.75% level of dietary levan supplementation.     

Effect of dietary sodium butyrate on growth performance,enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus

The present study was conducted to investigate the effects of dietary sodium butyrate on growth performance, intestine enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus. A basal diet was supplemented with sodium butyrate at 0.0 (control), 0.25, 0.5, 1, 2 and 4 g/kg feed for 8 weeks. The final body weight, weight gain (WG), specific growth rate (SGR) and condition factor (CF) increased with increasing dietary sodium butyrate up to 2.0 g/kg, and thereafter declined, while feed conversion ratio exhibited an opposite trend. Compared with the control, the 2.0 g/kg sodium butyrate group had higher condition factor (CF) significantly (p < 0.05). Whole body ash decreased with increasing dietary sodium butyrate level, with the lowest whole body ash content in 4.0 g/kg sodium butyrate (p < 0.05). Compared with the control, the 2.0 g/kg sodium butyrate group had significantly increased plasma glucose, cholesterol, albumin level, A/G ratio, ALT and AST contents (p < 0.05), while significantly decreased plasma ALT/AST ratio (p < 0.05). As for the intestinal digestive and brush border enzymes activities, compared with the control, the 2.0 and 4.0 g/kg sodium butyrate groups had significantly increased intestinal protease, amylase, AKP and Na⁺‐K⁺‐ATPase activities (p < 0.05), respectively. The relative level of intestinal CDX2 mRNA of fish significantly increased with dietary sodium butyrate level. Compared with the control, the 2.0 g/kg sodium butyrate groups had significantly increased the expression of intestinal CDX2 and CREB mRNA (p < 0.05), respectively. In conclusion, these results suggested that the optimum sodium butyrate level for juvenile golden pompano could be 2.0 g/kg of the diet.     

Effect of dietary vitamin B6 supplementation on growth and intestinal microflora of juvenile golden pompano (Trachinotus ovatus)

In order to investigate the dietary vitamin B₆ (VB₆) requirement for juvenile golden pompano, an experiment of six different diets with six dietary VB₆ levels (0, 3.75, 7.47, 10.6, 13.7 and 18.5 mg/kg) was conducted. The results indicated that the content of dietary VB₆ significantly increased weight gain rate (WGR), specific growth rate (SGR) and feed conversion ratio (FCR; p < 0.05). Golden pompano fed diet containing 7.47 mg/kg VB₆ achieved the maximum ALT, AST, GPx, hepatic VB₆ concentrations as well as the minimum MDA, however, the ALT, AST, GPx and hepatic VB₆ concentrations decreased while MDA increased with a further increase in dieatry VB₆. The highest values of GR and POD occurred at the 10.6 and 13.7 mg/kg dietary VB₆ (p < 0.05) respectively. A diet supplemented with 7.47 mg/kg VB₆ increased intestinal Na⁺, K⁺‐ATPase, Chymotrypsin, γ‐glutamyl transpeptidase, creatine kinase and amylase activities (p < 0.05). The relative abundance of Tenericutes and Bacteroidetes decreased while Proteobacteria and Firmicutes improved with an increase in dietary VB₆ levels up to 7.47 mg/kg. Quadratic regression analysis on WGR, hepatic VB₆ concentrations, AST and ALT indicated that the optimum dietary VB₆ levels for juvenile golden pompano were 8.84–9.28 mg/kg.     

Effects of dietary fish oil substitution with linseed oil on growth,muscle fatty acid and metabolism of tilapia (Oreochromis niloticus)

This study was conducted to evaluate the effect of linseed oil (LO) replacing different levels of fish oil (FO) on growth, muscle fatty acid composition and metabolism of gift tilapia (Oreochromis niloticus) (mean initial weight 22 ± 0.5 g) in indoor recycle aquarium tanks for 8 weeks. Fish fed the diet with 50% of the oil as LO had higher final body weight (FWG), specific growth rate (SGR) and protein efficiency ratio (PER) than the other groups (P < 0.05). Hepatopancreas lipid content of fish fed 50% LO was lower than the other groups. Total n‐3 and n‐6 PUFA contents in the dorsal muscle and superoxide dismutase (SOD) activity in serum increased with increasing dietary LO level. Fish fed 50% LO had higher alanine transaminase (ALT), aspartate transaminase (AST) and lipoprotein lipase (LPL) activities in hepatopancreas and total antioxidant capacity (T‐AOC) and alkaline phosphatase (AKP) activities in serum than the other groups (P < 0.05). However, malate dehydrogenase (MDH) activities and malondialdehyde (MDA) contents in hepatopancreas were lower than other groups (P < 0.05) with a 50% substitution level. Results of this study indicated that LO could substitute <50% FO without influencing the growth of tilapia. The higher substitution levels of LO induced negative influences on growth, feed utilization and antioxidant ability of tilapia, but could promote DHA synthesis in tilapia muscle.     

Effects of butaphosphan and cyanocobalamin combination on plasma immune and biochemical parameters of Olive flounder (Paralichthys olivaceus) subjected to crowding stress

The study evaluated the effects of intramuscular injection of butaphosphan and cyanocobalamin combination (BPC) on the plasma levels of cortisol, cytokines, and the activities of selected enzymes in Oliveflounder exposed to crowding stress. Three groups of fish (n = 12 per group) were kept in different glass tanks with stocking densities of 46 kg/m³(group BP and SCH) and 15 kg/m³ (group SCL). Group BP was treated with 0.5 ml of BPC. While groupSCH and SCL were treated with 0.5 ml of saline. Blood was collected a day after injection (T1), and weekly for two consecutive weeks (T2 and T3). The plasma levels of interleukin (IL) ‐1β, IL‐6, cortisol, lysozyme, and the activities of aspartate aminotransferase (AST) and alanine transaminase (ALT)were determined using an ELISA kit. The statistical significance of differences was assessed using ANOVAfollowed by Tukey‐HSD test (P < 0.05 considered significant).A significantbody weight gain was measured in the BP and SCL groupscompared with SCH at T3. The activities of AST (at T1, T2, and T3) and ALT (at T1 and T2) were declined significantly (P < 0.05) following treatment of overcrowded fish with BPC than saline. Similarly, BPCcaused a significant reduction in the levels of cortisol and IL‐1β at T2 and T3.This is the first report of the effect of the combination of butaphosphan and cyanocobalamin on plasma immune and biochemical parametersin fish. Therefore, the combination could be beneficial in preventing overcrowding‐induced immune suppression and tissue damage in Olive flounder.     

Effects of dietary mannanase on growth,metabolism and non‐specific immunity of Tilapia (Oreochromis niloticus)

An experiment was designed to assess the effects of a commercial β‐mannanase on performance and immunity of tilapia fed plant‐based diets. A basal diet was supplemented with 0.0 (control), 0.5 and 1.0 g β‐mannase kg⁻¹ to formulate three experimental diets. Each treatment contained 4 tanks with 30 fish per tank. Trial lasted 8 weeks. Our results demonstrated that β‐mannanase addition (0.5 and 1.0 g kg⁻¹) improved significantly (P < 0.05) the final weight, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) compared with the control (0.0 g kg⁻¹). There were no significant differences in feed intake (FI) and survival rate (SR) among the 3 dietary treatments (P > 0.05). β‐mannanase supplementation also led to an increase (P < 0.05) in amylase, trypsin and Na⁺K⁺‐ATPase activities in intestine, and an decrease (P < 0.05) in aspartate transaminase (AST) and alanine transaminase (ALT) activities in serum compared with the control. However, dietary enzyme supplementation had no significant effect on the serum triacylglycerol (TG), cholesterol (CHO), high density lipoprotein cholesterol (HDL‐C), low density lipoprotein cholesterol (LDL‐C) and very low density lipoprotein cholesterol (VLDL‐C) (P > 0.05). Moreover, the dietary β‐mannanase supplementation groups exhibited an increase in the total leukocyte counts (WBC), differential leukocyte counts, respiratory burst activity, lysozyme activity and superoxide dismutase (SOD) activity compared to the controls (P < 0.05). In conclusion, β‐mannanase addition to tilapia diets improved feed utilization and non‐specific immunity resulting in improvements in growth performance.