Efficacy of nutritional selenium nanoparticles on growth performance, immune response, antioxidant capacity, expression of growth and immune-related genes, and post-stress recovery in juvenile Sobaity seabream (Sparidentex hasta).

This study evaluated the impacts of nano-Se on the growth, immunity, antioxidant capacity, physiological parameters, gene expression, and stress resistance of fingerling Sobaity seabream (Sparidentex hasta). The fish with an average weight of 21.5 ± 0.1 g were divided into four treatment groups in triplicates that received one of the test diets supplemented with varying levels of nano-Se: 0 (control), 0.5 (Se-0.5), 1 (Se-1), and 2 (Se-2) mg/Kg for 60 days. The results showed that final weight, weight gain rate, specific growth rate, feed intake, and feed conversion ratio improved with significant linear and quadratic trends (P < 0.05) in response to nano-Se-supplemented diets, and the best values were measured in the Se-2 group. Superoxide dismutase activity level remained unaffected among the four groups (P > 0.05). Catalase activity increased in nano-Se-supplemented groups, with the highest level measured in fish fed the Se-0.5 diet. Glutathione peroxidase activity levels were not significantly different between the control and nano-Se groups, but the lowest malondialdehyde concentration was detected in the Se-2 group. Nano-Se had no marked effect on total plasma Ig levels; however, the highest lysozyme activity and alternative complement activity (ACH50) were observed in the Se-0.5 and Se-2 groups, respectively. No significant differences (P > 0.05) were observed in plasma total protein, albumin, globulin, triglyceride, and thyroid hormone (T3 and T4) contents among the groups. However, the lowest cholesterol and low-density lipoprotein values and the highest high-density lipoprotein concentration were measured in the Se-2 group. The Se-0.5 and Se-1 groups exhibited significantly lower levels of aspartate aminotransferase activity, and the lowest alkaline phosphatase activity level was detected in the Se-1 group. The expression level of insulin-like growth factor I gene in all nano-Se-fed groups was significantly higher than the control. Also, the expression of interleukin-1ß and lysozyme genes was significantly upregulated in nano-Se-supplemented groups, with the highest values in the Se-2 group. Following acute crowding stress, plasma cortisol and lactate levels at all post-stress time intervals were not significantly different among the experimental groups. Fish fed the Se-0.5 and Se-2 diets tended to have lower plasma glucose concentrations than other groups. In conclusion, dietary nano-Se at 2 mg/kg is recommended to promote growth performance and enhance antioxidant and immune parameters in Sobaity juveniles.

Dietary Selenium Nanoparticles Improved Growth and Health Indices in Asian Seabass (Lates calcarifer) Juveniles Reared in High Saline Water.

A 60-day study was carried out to determine the effect of dietary selenium nanoparticles (SeNP) on growth, digestive enzymes, and health status of Asian seabass (Lates calcarifer, 46.5 ± 0.2 g) juveniles reared in high saline water (48 ppt). Five levels of SeNP were added to a basal diet (45% protein, 15% lipid), including control (0), 0.5 (SeNP0.5), 1.0 (SeNP1), 2 (SeNP2), and 4 (SeNP4) mg SeNP kg-1 diet. Fish were stocked into fifteen 2,000 L tanks (50 fish tank-1) filled with 1,800 L sand-filtered seawater (26.5 ± 1.5°C, 48.0 ± 0.2 ppt) in a flow-through system. Each dietary treatment was performed in three replicates. The growth rate positively increased in both linear and quadratic trends with increasing dietary SeNP level (P < 0.05). The liver Se concentration increased with increasing SeNP in diet (P < 0.05). Gut total protease, trypsin, chymotrypsin, alkaline phosphatase (ALP), lipase, and α-amylase activities were significantly enhanced in the SeNP4 group compared to the other treatments (P < 0.05). Antioxidant capacity improved in fish-fed SeNP2 and SeNP4 diets regarding catalase and superoxide dismutase activities and the liver glutathione content. Serum lysozyme and hemolytic activities and white blood cells' respiratory burst activity in the control were lower than in fish-fed SeNP-supplemented diets. Serum total protein, globulin, and globulin/albumin ratio in fish-fed SeNP1, SeNP2, and SeNP4 diets were higher than the other groups (P < 0.05). The interleukin-10 and granulocyte-macrophage colony-stimulating factor genes' relative transcription levels in the gut of fish-fed SeNP4 were higher than the other groups. Serum cholesterol, triglycerides, ALP, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase significantly decreased in fish with increasing SeNP content in the diet. In conclusion, supplementing the diet with 4 mg kg-1, SeNP was recommended to improve growth and health indices in L. calcarifer juveniles reared in high saline water (48 ppt).

Effect of killed autogenous polyvalent vaccines against Vibrio harveyi, V. alginolyticus and Streptococcus iniae on survival and immunogenicity of Asian seabass (Latescalcarifer).

Vibriosis and Streptococcosis are the most important bacterial diseases that infect Asian seabass (Lates calcarifer) in various stages of its life cycle. Vaccination is a cost-effective strategy to prevent the occurrence of infectious diseases and increase sustainability in the aquaculture industry. This study was aimed to develop and evaluate a killed polyvalent vaccine against Vibrio harveyi, V. alginolyticus and Streptococcus iniae, delivered by intraperitoneal injection in Asian seabass. The fish were divided into three groups with 60 fish in triplicate: I) a control group injected with phosphate-buffered saline (PBS), II) a group vaccinated by polyvalent vaccine (V. alginolyticus + V. harveyi + S. iniae) and III) a group vaccinated with the same polyvalent vaccine plus an oral booster. Immunological parameters and antibody titer were measured before and at three, five-, and eight-weeks post-vaccination. The efficacy of the killed vaccine was assessed five weeks post-vaccination by challenging with each isolate separately. The vaccinated groups had higher survival rate than control group. The highest relative percentage survival rate, 85.71 ± 3.57 % was observed in group III when challenged with V. harveyi. The vaccinated fish produced significantly higher antibody titers against V. alginolyticus, V. harveyi and S. iniae than the control group (P < 0.05). Non-specific immune parameters were significantly enhanced in the vaccinated groups, especially group III, compared to the control. The results demonstrated that the administration of a killed polyvalent vaccine can effectively protect Asian seabass against V. alginolyticus, V. harveyi and S. iniae.

Larval rearing and ontogeny of digestive enzyme activities in yellowfin seabream (Acanthopagrus latus, Houttuyn 1782).

The present research was conducted to provide insight into digestive larval capacity in Acanthopagrus latus larvae from hatching up to 30 days after hatching (DAH). Newly hatched larvae were stocked into six 300-L cylindrical polyethylene tanks at a density of larvae 50 larvae/L and reared by means of the green water system using Nannochloropsis oculata (0.5 × 106/mL). After mouth opening, larvae were fed with rotifers (5-16 individual/mL) from 2 to 20 DAH; then, Artemia nauplii (0.5-3.0 individuals/mL) were offered to larvae from 18 to 30 DAH, meanwhile a commercial microdiet was offered to larvae from 25 to 30 DAH. Larval performance in terms of growth and survival, and the assessment of the activity of selected digestive enzymes ontogeny of digestive enzymes activities was evaluated in larvae sampled at 0 (hatching), 7, 15, 22 and 30 DAH. Larvae showed an exponential growth characterized by two different growth stanzas, a first one characterized by slow growth rates comprised between hatching to 15 DAH (4.7 ± 0.2 mm), followed by a period of faster growth rates between 16 and 30 DAH (7.5 ± 0.6 mm). The activities of the brush border (alkaline phosphatase, ALP) and cytosolic (leucine-alanine peptidase, LAP) enzymes, as well as those of the pancreatic ones like total alkaline proteases, bile salt-activated lipase and α-amylase were detected from the mouth opening stage. Total activities of pancreatic and gastric enzymes increased with larval growth showing an enhancement of digestive capacities with larval age and size. The intestinal maturation in A. latus as assessed by the ratio of AP to LAP did not occur as expected by end of the first month of life suggesting the complete establishment of digestive luminal processes may take place at older ages. This study related to the growth patterns and ontogenic changes in activity of pancreatic, gastric and intestinal enzymes in A. latus and their nutritional regulation may be considered as the first step for improving the larviculture, as well as assessing and refining the nutritional requirements during the larval and early juvenile stages of this sparid species.

Effects of dietary fern (Adiantum capillus-veneris) leaves powder on serum and mucus antioxidant defence, immunological responses, antimicrobial activity and growth performance of common carp (Cyprinus carpio) juveniles.

A 56-day research was performed to examine the influence of graded levels (0 (control), 0.5, 1 and 2%) of Fern (Adiantum capillus-veneris) leaves powder (FLP) in diet on immune competence and growth of common carp (Cyprinus carpio, initial weight = 20 g). The serum total immunoglobulins content and lysozyme activity in the 1 and 2% FLP groups remarkably increased compared to the other groups (P < 0.05). The skin mucosal lysozyme activity enhanced with increasing dietary FLP level in a dose-response manner. Fish fed on the FLP-supplemented diets had higher skin-mucosal superoxide dismutase activity than the control (P < 0.05). However, serum antioxidant enzymes were not affected by dietary fern (P > 0.05). The serum bactericidal activity against human and fish pathogens increased with enhancing the FLP level in diet against Staphylococcus aureus, Escherichia coli (EHEC ATCC 43895), Escherichia coli (CI), Pseudomonas aeruginosa, Klebsiella pneumonia and Aeromonas hydrophila. The serum antibacterial activity against Yersinia ruckeri in the 2% FLP group was higher than the other treatments. Furthermore, the serum bactericidal activity against P. aeruginosa (ATCC 27853) only observed in fish fed on the 1 and 2% FLP-supplemented diets. The skin mucosal bactericidal activity and inhibitory effects increased with enhancing the FLP level in diet against E. coli, K. pneumonia, Y. ruckeri and A. hydrophila in a dose response manner. Moreover, the skin mucosal bactericidal activity against S. aureus only observed in fish fed on 1 and 2% FLP-supplemented diets. The weight gain values in the 1 and 2% FLP groups were higher than the other treatments (P < 0.05). Feed conversion ratio (FCR) improved with increasing FLP level in diet in a dose-response manner (P < 0.05). By considering serum and mucosal bactericidal activities against different pathogenic bacteria, the supplementation of 2% FLP in diet is recommended for C. carpio during grow-out phase.

Humoral and skin mucosal immune parameters, intestinal immune related genes expression and antioxidant defense in rainbow trout (Oncorhynchus mykiss) fed olive (Olea europea L.) waste.

A six-week feeding trial was carried out to evaluate the effects of inclusion of dietary olive waste cake (OWC, 0, 0.5, 2.5 and 5 g kg-1 diet) on performance, antioxidant condition and immune responses of rainbow trout (Oncorhynchus mykiss) (2.5 ± 0.1 g). Supplementing diet with 2.5 and 5 g OWC kg-1 diet significantly enhanced serum and mucosal lysozyme activity in fish. Regarding mucosal immunity, fish fed 2.5 and 5 g OWC kg-1 diets had higher skin mucus total Ig concentrations than other groups. In relation to antioxidant status, those in 2.5 g OWC kg-1 and the control groups exhibited the highest and the least liver superoxide dismutase and glutathione peroxidase activities, respectively. Furthermore, the activity of liver glutathione S transferase in fish fed 2.5 and 5 g OWC kg-1 diets was higher than the other treatments. In respect to gut cytokines gene expression, our findings demonstrated dietary OWC did not influence interlukines-1ß and 10 genes expression, but relative expression of IL8 gene gradually up-regulated with increasing dietary OWC level. Moreover, fish fed 0.5 g OWC kg-1 and the control diets had the highest and the lowest gut tumor necrosis factor-α gene expression values, respectively. The relative expression of transforming growth factor-ß significantly down-regulated in gut of fish fed 2.5 and 5 g OWC kg-1 diets compared to other groups. Supplementing diet with OWC pronouncedly improved growth and feed conversion ratio in fish compared to the control. Overall, the findings of this study suggested that inclusion of 2.5 g OWC kg-1 diet can improve growth rate, oxidative stress status, humoral and skin mucosal immune responses in O. mykiss fingerlings and it can be considered as a functional feed additive for this species.

Enhanced mucosal immune responses, immune related genes and growth performance in common carp (Cyprinus carpio) juveniles fed dietary Pediococcus acidilactici MA18/5M and raffinose.

The present study was conducted to evaluate the effects of dietary Pediococcus acidilactici (PA) and raffinose (RF) alone or in combination on growth performance, mucosal immune factors and immune related genes expression in common carp (Cyprinus carpio) juveniles. Fish with initial weight of 10.0 ±â€¯2.5 g were fed the following experimental diets for 60 days: control (without supplementation), prebiotic (2 g RF kg-1 diet), probiotic (6 × 108 CFU g-1PA) and synbiotic (2 g RF kg-1+ 6 × 108 CFU g-1PA). Carp fed synbiotic and probiotic diets had the highest (19.53 ±â€¯0.16) and the lowest (18.05 ±â€¯0.65) final body weight, respectively and the other experimental groups showed intermediate values. Singular administration of PA or in combination with 2 g RF kg-1 significantly increased skin mucus total immunoglobulin (Ig) and protein compared other groups, meanwhile, values of skin mucus protease activity enhanced by dietary immunostimulants administration in comparison with the control (P < 0.05). The expression of gene encoding lysozyme in skin pronouncedly increased by supplementing diets with singular or mixture of PA and RF; however, the expression of intestinal lysozyme gene as well as tumor necrosis factor-α genes expression in skin and intestine were not affected by administrating different immunostimulants (P > 0.05). The highest growth performance was noticed in fish fed synbiotic (P < 0.05). Overall, the combination of 2 g RF kg-1 with 6 × 108 CFU g-1PA is recommended for improving immunological responses of C. carpio juveniles.

Enrichment of rainbow trout (Oncorhynchus mykiss) fingerlings diet with microbial lysozyme: Effects on growth performance, serum and skin mucus immune parameters.

A two-month study was conducted to determine the influence of different levels of microbial lysozyme (LZ) contents (0, 0.5, 1.0, and 1.5 g kg-1 of diet) on growth performance, serum and skin mucus immune parameters as well as intestinal immune-related genes expression in rainbow trout Oncorhynchus mykiss fingerlings (5.5 ±â€¯0.1 g). Growth performance and feed utilization were not affected significantly by dietary LZ. Fish fed LZ-supplemented diets had higher serum total immunoglobulins concentration than the control group. In addition, fish fed 1.5 g LZ kg-1 diet had the highest skin mucosal total protein and immunoglobulin contents compared to other experimental groups. Furthermore, skin mucosal lysozyme and alkaline phosphatase activities as well as intestinal tumor necrosis factor-α and interlukine-1ß relative genes expression were higher in fish fed 1.0 and 1.5 g LZ kg-1 diets than the other groups. Overall, the present results clearly showed that LZ powder can be considered as a potential immunostimulant in O. mykiss fingerlings, but in the long term period it may result in negative effects on intestinal health as a consequence of inducing pro-inflammatory cytokines gene expression in the intestine.

Combined effects of dietary low molecular weight sodium alginate and Pediococcus acidilactici MA18/5M on growth performance, haematological and innate immune responses of Asian sea bass (Lates calcalifer) juveniles.

Six weeks feeding trial was conducted to examine the effects of dietary administration of low molecular weight sodium alginate (LMWSA) and Pediococcus acidilactici MA 18/5M (PA) on humoral and mucosal immune responses, haematological parameters and growth performance, of Lates calcarifer juveniles. Fish (12.0 ±â€¯0.2 g) were fed experimental diets as follows: Control (diet 1, basal diet), 5 g kg-1 LMWSA (diet 2), 10 g kg-1 LMWSA (diet 3), 0.9 × 107 CFU g-1 PA (diet 4), 5 g kg-1 LMWSA + 0.9 × 107 CFU g-1 PA (Diet 5), and 10 g kg-1 LMWSA + 0.9 × 107 CFU g-1 PA (Diet 6). Results indicated a significant (P < 0.05) increase in innate immune parameters including serum lysozyme, bactericidal, hemolytic and respiratory burst activities as well as mucosal immune responses including lysozyme and bactericidal activities, when diet was supplemented with immunostimulants. Moreover, the combined effects of LMWSA with PA resulted in more pronounced immunological responses compared to the control and singular administration. Red and white blood cell counts significantly increased with either singular or combined administration of LMWSA and PA compared with the control group (P < 0.05). The singular administration of PA and combined supplementation of 5 g kg-1 LMWSA with PA significantly increased growth performance and feed intake compared with other experimental groups (P < 0.05). These results indicated that combined administration of LMWSA and PA can be considered as beneficial feed additive and immunostimulant in L. calcarifer juveniles.