Moderate salinities enhance growth performance of Nile tilapia (Oreochromis niloticus) fingerlings in the biofloc system

The biofloc technology (BFT) has recently gained attention as an economic and environmentally sustainable system for aquaculture. The use of BFT with salinized water could be used to minimize the negative effects of nitrogenous waste and improve the growth performance of Nile tilapia. This work evaluated the growth performance, survival, gill lesions, and fillet composition of tilapia fingerlings (Oreochromis niloticus) reared during 70 days with different salinities (0, 4, 8, 12, and 16 g/L) in a biofloc previously developed. The results indicated that the use of mature bioflocs alone was sufficient to avoid fingerling mortality due to nitrite peaks. Moreover, the salinized water, especially between 4 and 8 g/L (maximum points around 6 g/L), can be recommended in BFT to improve the growth performance of tilapia in the initial culture phase. The salinity level that was evaluated did not affect fillet composition nor the occurrence of gill lesions. The total ammonia nitrogen concentration was lower in freshwater than in salinized water (p < 0.05). Nitrite peaks in salinities of 12 and 16 g/L appeared later and were higher in comparison to other treatments. Considering minimal water exchange in BFT, the use of salinized water in this system may be environmentally viable.     

Alternate weekly exchanges of feeding regime affect the diversity of intestinal microbiota and immune status of Nile tilapia Oreochromis niloticus

The intestinal microbiota play an important role in the maintenance of the fish health. In this study, 150 Oreochromis niloticus were divided into five groups (G1–G5) and were subjected to alternate weekly exchange of feeding rate and frequency (3, 3, 1.5, 1.5 and 0%) and (2, 1, 2, 1 and 0 time), respectively, for 8 weeks. Enumeration of total intestinal bacteria revealed that the abundance of Lactobacillus was negatively correlated with those of Aeromonas, Pseudomonas and Edwardsiella. The abundance and proportion of Lactobacillus were affected by the change in feeding frequency. When fish were infected with Aeromonas hydrophila, survival rate significantly decreased in G5 (starved fish) compared with G1 (control group). The pro‐inflammatory cytokines, interleukin (IL)‐1β and tumour necrosis factor‐α, and anti‐inflammatory cytokine IL‐10 as well as serum antibacterial activity and respiratory burst activity were positively correlated with the proportion of Lactobacillus. The alternate weekly exchange of feeding regime did not alter the parametric morphology of the intestine, height and width of intestinal villi, number of goblet cells and width of muscle layer, except in G5. In conclusion, the alternate weekly exchange of feeding regime to starvation (G5), decrease in the feeding frequency to 1 time/day (G3) and decrease in feeding rate to 1.5% (G4) suppressed the immune status of fish, which became vulnerable to bacterial infection. Immune suppression was positively correlated with a decreased proportion of Lactobacillus spp.     

Combined effects of temperature and salinity on yolk utilization in Nile tilapia (Oreochromis niloticus)

The combined effects of temperature and salinity on the yolk utilization of sac fry in Nile tilapia (Oreochromis niloticus) were investigated using central composite experimental design and response surface approach. Based on the preliminary trials, temperature was determined to range from 22 to 34°C, and salinity ranging from 2 ppt to 10 ppt. The utilization was mensurated in terms of yolk sac volume. Results showed that the linear effects of temperature and salinity on the yolk utilization was significant (P < 0.01); the quadratic effects of and the interaction between the two factors were significant (P < 0.05); temperature was more important than salinity in influencing the yolk utilization. The model equation of yolk sac volume towards temperature and salinity was established. From those high R² values, the model had excellent goodness of fit to experimental data and could be applied for predictive purpose. What with the production cost, it is suggested that the temperature/salinity combination, i.e. 28–30°C/4–6 ppt, be employed during the period of sac fry rearing, in which the yolk utilization was on average 98.6%.     

Feed intake,growth and metabolism of Nile tilapia (Oreochromis niloticus) in relation to dissolved oxygen concentration

The objectives of the present study were to determine, for Nile tilapia of different body weights and fed to satiation, (1) the incipient dissolved oxygen (DO) concentration at which feed intake starts to level off and (2) the effect of DO on nitrogen and energy balances. Two successive experiments were conducted with two weight classes of male Nile tilapia (>200 g, Experiment 1, and <100 g, Experiment 2). Twelve aquaria were assigned to four DO levels in each experiment. Fish were fed to apparent satiation for 16 and 20 days in Experiments 1 and 2 respectively. The DO‐feed intake curve of fish<100 g appeared to level off at a DO of 3 mg L^?1, while that of fish>200 g continued to increase as DO increased from 2.6 to 6.0 mg L^?1. The latter curve suggests that the incipient DO for fish>200 g is about 5.5 mg L^?1. Fish tended to reduce the energy requirement for maintenance as DO declined. A DO reduction caused an increase in the apparent digestibility coefficient (ADC), but further declines in DO to levels below a critical level decreased ADC. The DO did not have a considerable effect on the metabolizable energy: digestible energy ratio of the diet.     

CD40 induces an antimicrobial response against the intracellular pathogen Streptococcus agalactiae in Nile tilapia,Oreochromis niloticus

Streptococcus agalactiae is a Gram‐positive facultative intracellular bacterium that leads to severe economic loss of tilapia worldwide. Previous studies demonstrated that CD40 contributes to host protection against intracellular injection. In this study, CD40 was characterized from Nile tilapia (Oreochromis niloticus), named OnCD40. Sequence analysis showed that open reading frame of OnCD40 was 933 bp, containing a single peptide, a transmembrane domain and four cysteine‐rich domains. The qRT‐PCR revealed that OnCD40 was expressed in all examined tissues with the most abundant ones in spleen and thymus. After S. agalactiae stimulation, the expression of OnCD40 was significantly induced in most of the detected organs. Moreover, OnCD40‐overexpressing fish elicited significant protection against subsequent S. agalactiae challenge; approximately 10000‐fold fewer bacteria were detected in spleen of OnCD40‐overexpressing fish in comparison with control fish. Thus, CD40 had protecting function in Nile tilapia against intracellular pathogens.     

Vitamin A affects haematology,growth and immune response of Nile tilapia (Oreochromis niloticus,L.), but has no protective effect against bacterial challenge or cold‐induced stress

Vitamin A (vitA) is an essential nutrient that acts as an endocrine regulator of several metabolic pathways, modulating normal growth and health status of animals. Although the importance of vitA for normal haematology and immune response is well documented for higher vertebrates, there is limited information on the physiological effects of vitA on fish. Therefore, we designed a 130‐day feeding trial to evaluate the effect of vitA supplementation on growth, haematology, immune function and resistance to experimental infection with Aeromonas hydrophila and cold‐induced stress. A group of 320 Nile tilapia fingerlings 7.49 ± 0.19 g weight (mean ± SD) were randomly stocked into 40 250 L‐aquaria and fed practical diets containing graded levels of vitA (0, 0.06, 0.12, 0.24, 0.48, 0.96, 1.92, 3.84 mg retinol (ROH) kg^?1 diet. Growth, haematology, plasma protein profile and immune response were significantly affected by vitA supplementation; however, no clear protective effect of vitA supplementation on disease and cold stress resistance were observed in this study. Clinical signs of vitA deficiency were: resting and abnormal swimming behaviour, exophthalmia, haemorrhages at the base of fins and on skin, serous fluids in abdominal cavity, neutropenia, reduction in red blood cell count, haematocrit and haemoglobin evolving to high mortality rates in a short period of time. A dietary level of vitA around 1.2 mg ROH kg^?1 may be required to prevent gross deficiency signs and promote proper growth and health status of Nile tilapia. VitA does not seem to have a pronounced effect on leucocyte differentiation, but clearly plays an important role on maintaining normal erythropoiesis.     

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.     

Growth performance,oxidative stress indices and hepatic carbohydrate metabolic enzymes activities of juvenile Nile tilapia,Oreochromis niloticus L., in response to dietary starch to protein ratios

The effect of various dietary starch to proteins ratios (STA/P) on growth performance, oxidative status and liver enzyme activities involved in intermediary metabolism in juvenile Nile tilapia was evaluated. Four isocaloric‐practical diets (12.73 MJ kg^?1 digestible energy) with increasing STA/CP ratios were formulated. These were designated D₀ (344 g crude protein (CP) and 163.5 g starch (STA) kg^?1), D₁ (310 g CP and 243 g STA kg^?1), D₂ (258 g CP and 322 g STA kg^?1) and D₃ (214 g CP and 401 g STA kg^?1). Each diet was fed to triplicate groups of 60 fish (2.7 g) for 45 days. Compared with the control diet (D₀), significantly (P < 0.05) depressed growth and feed efficiency were observed only in the groups fed on diet D₃. The activities of hepatic enzymes involved in glycolysis and lipogenesis pathways were significantly enhanced in groups fed on diet D₃ compared with other diets. A significant (P < 0.05) increase in catalase activity was detected only in groups fed on diet D₃. Similarly, a significant (P < 0.05) enhancement in superoxyde dismutase, glutathione S‐transferases and glutathione peroxidise was observed in groups fed on diets D₂ and D₃ compared with other diets. Results demonstrate the ability of juvenile Nile tilapia to spare protein by dietary carbohydrate.     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

Dualistic feeding pattern of Nile tilapia (Oreochromis niloticus,L.) reared under different self‐feeding system conditions

Nile tilapia (Oreochromis niloticus) is one of the most important cultured species worldwide. The research aims to clarify the feeding characteristics, such as daily feeding pattern of Nile tilapia using a self‐feeding technique. The feeding pattern was conducted under two rearing conditions, indoor treatments under a controlled light regime (LD 12:12) and constant water temperature (25°C), and outdoor treatments under natural conditions which consisted of duplicate trials with two periods each. The outdoor treatment was carried out from early summer through late autumn in Mie, Central Japan. Daily self‐feeding activity of Nile tilapia in indoor treatments was nearly daytime feeding pattern, synchronizing with the given photoperiod (24 hr). However, the self‐feeding activity in the outdoor experiments from early summer to early autumn was almost daytime feeding pattern, but it became less clear and shifted to a nighttime feeding profile in late autumn. The results revealed that Nile tilapia has a dualistic capacity for demand‐feeding both in light and dark phases. These results might have been caused by the seasonal change in light intensity and/or water temperature. Information obtained from the self‐feeding experiments enables us to identify the influence of environmental changes on the physiological condition of farmed fish through their expression of appetite.     

Effects of putative dietary probiotics from the gut of milkfish (Chanos chanos) on the growth performance and intestinal enzymatic activities of juvenile Nile tilapia (Oreochromis niloticus)

Enzymes synthesized by the gut microbiome are recognized as vital for assisting host metabolism. This study was conducted to screen and identify bacterial isolates with enzyme-producing abilities from the intestine of milkfish Chanos chanos. Overall, 114 isolates were collected, of which 58% showed clear zones indicating various enzyme activities via plate-based assays. Specifically, 65 isolates were amylolytic, 1 isolate was cellulolytic, and 18 isolates exhibited both activities. The two most promising isolates—CC8 and CC27—were further subjected to spectrophotometric enzyme assays to determine their amylase, cellulase and protease activities. Results showed that amylase, cellulase and protease activities (6.00 ± 0.01, 0.52 ± 0.02 and 18.62 ± 0.16 U mg^?1 protein, respectively) were higher in CC8 than in CC27. Using 16S rRNA gene sequencing, the isolates were putatively identified as Vibrio sp. and Bacillus cereus respectively. At a suspension (mL) to feed (g) ratio of 3:25, dietary supplementation of putative probiotics was carried out to evaluate the growth performance, survival and enzymatic activities of juvenile Nile tilapia Oreochromis niloticus. Results revealed that probiotic-supplemented fish exhibited better growth performance—final average body weight, weight gain, specific growth rate and feed conversion ratio—along with increased intestinal enzymatic activities compared with the control (p < 0.05). Survival rates ranged from 80% to 90% and were statistically similar in all treatments (p > 0.05). This study concludes that the application of Vibrio sp. CC8 and Bacillus cereus CC27, supplemented either in monostrain or multistrain preparation, can promote growth and enzymatic activities in juvenile Nile tilapia.     

Beneficial effects on growth,haematic indicators,immune status,antioxidant function and gut health in Juvenile Nile tilapia (Oreochromis niloticus) by dietary administration of a multi‐strain probiotic

The present study was conducted to administer a commercial multi‐strain probiotic (MP), Yilibao (Bacillus velezensis:Bacillus cereus:Lactobacillus casei = 2:2:1), in Nile tilapia, Oreochromis niloticus. In terms of aerobic Bacillus spp. counts, we produced five diets containing 0, 0.34, 1.68, 3.36 and 6.72 g/kg of MP dry product (Control, T1, T2, T3, T4). Seven hundred and fifty tilapia juveniles (13.26 ± 0.01 g) distributed into 25 tanks in five replications were fed their diet for eight weeks. Results showed that fish fed T3 and T4 diets displayed significantly higher final body weight and weight gain (p < .05). Further, all MP‐treated fish exhibited remarkably decreased plasma lipid profiles (cholesterol, triglycerides) (p < .05) and fish fed T3 and T4 diets displayed significantly higher plasma myeloperoxidase activity and complement C3 content (p < .05). Additionally, fish fed T3 and T4 diets exhibited significantly promoted total antioxidant capacity, glutathione peroxidase activity, glutathione level and declined malondialdehyde content in the plasma and/or liver (p < .05). Moreover, significantly elevated chymotrypsin activity, villus height and intraepithelial lymphocytes counts were found in fish fed T3 and T4 diets (p < .05). As fish fed T3 and T4 diets surpassed the other treatments in growth, immune–antioxidative status and gut health, the recommended dose for tilapia is 3.36–6.72 g/kg.     

Effects of ferulic acid on growth performance,immunity and antioxidant status in genetically improved farmed tilapia (Oreochromis niloticus) fed oxidized fish oil

A study was conducted to characterize the effects of dietary oxidized fish oil on the growth performance, immunity and antioxidant status of genetically improved farmed tilapia (Oreochromis niloticus) and to determine the role of ferulic acid on the oxidative damage induced by the oxidized fish oil. The tilapia (13.73 ± 0.31 g) were fed four experimental diets containing untreated (peroxide value, POV: 2.2 meq/kg) and highly oxidized (POV: 120.6 meq/kg) fish oil either with or without ferulic acid (0 or 400 mg/kg) supplementation for 12 weeks. From the results, the oxidized fish oil treatments increased antioxidant enzyme activities and MDA values but decreased the weight gain and the immunological parameters in tilapia. Meanwhile, the serum biochemical indices were significantly affected by the oxidized fish oil. Besides, the addition of ferulic acid partially counteracted the free radical‐induced damage and improved the health status of tilapia. In conclusion, the oxidized fish oil may induce oxidative stress, destroy liver, dysregulate lipid metabolism as well as reduce non‐specific immunity, and eventually result in growth inhibition of tilapia. The ferulic acid supplementation partially offset the negative effects of the oxidized fish oil on tilapia.     

Using stable isotope analysis to assess the relationship among dietary protein sources,growth, nutrient turnover and incorporation in Nile tilapia (Oreochromis niloticus)

Nile tilapia (1.20 g) were fed for 56 days in five treatments (FM100 containing only fish meal [FM] as a protein source; soybean meal (SBM) replace 25%, 50%, 75% and 100% of FM protein in the diets of FM75, FM50, FM25 and FM0, respectively) to assess the effects of dietary protein sources on growth, turnover half‐life of nitrogen and carbon stable isotopes (δ¹³C and δ¹⁵N) in muscle, isotope discrimination factors (Δ¹⁵N and Δ¹³C), and the relative contribution of protein sources to muscle growth. Results showed that the final body weight, weight gain and specific growth rate of fish fed diet FM25 were significantly higher than other treatments (p < .05). By day 35, fish under all treatments reached isotopic equilibrium, and the equilibrium values consistently increase as a function of stable isotope values in diets. Growth accounted for most isotopic changes in muscle and resulted in significantly shorter half‐lives of nitrogen and carbon in FM25 (p < .05). The Δ¹⁵N of FM25 and Δ¹³C of FM0 were significantly lower than other treatments (p < .05). The proportional contributions of SBM protein were significantly higher than their respective levels in FM75 and FM25 (p < .05), indicating a preferential incorporation of nitrogen from the SBM in muscle tissue.     

Relationship of RNA/DNA ratio to somatic growth of Nile tilapia juveniles (Oreochromis niloticus) under joint effects of temperature and salinity

Tilapia juveniles are a very important life stage, and reliably assessing their growth performance is of prime importance in aquaculture production. The suitability of the RNA/DNA as a bioindicator for growth evaluation in tilapia has not yet been reported. In this study, we examined suitability of RNA/DNA ratio for assessing growth of tilapia juveniles and variation in the ratio under the concurrent influences of temperature and salinity using the central composite design and response surface methodology. Results showed that under our experimental conditions, the synergistic effects of temperature and salinity on the RNA/DNA ratio were highly significant (P < 0.01), and the ratio varied with the two factors in a nonlinear fashion; the temperature × salinity interaction was detected at 1% significance level, and had a negative effect on the RNA/DNA ratio particularly at higher temperatures. Temperature was found to have a greater influence on RNA/DNA ratios than salinity. A model of the relation between RNA/DNA, temperature and salinity was developed, with an R² greater than 98%. The linear relationships of absolute growth, cumulative growth and specific growth of tilapia juveniles to RNA/DNA ratio all showed an R² of greater than 95%. Our results indicate that the RNA/DNA ratio was a responsive and accurate bioindicator for evaluating the growth performance of juvenile tilapia. These ratio‐based models are neither temperature‐ nor salinity‐dependent, and could be readily applicable to freshwater or brackish water aquaculture within temperature–salinity combinations ranging from 16 to 36°C and 0–16 ppt.     

The effect of dietary methionine concentrations on growth performance of juvenile Nile tilapia (Oreochromis niloticus) fed diets with two different digestible energy levels

A growth trial was conducted to examine the effect of dietary digestible energy (DE) content on methionine (Met) utilization and requirement in juvenile Nile tilapia (Oreochromis niloticus). Ten iso‐nitrogenous (288 g kg^?1 protein) practical diets, with two DE levels (10.9 MJ kg^?1; 12.4 MJ kg^?1) and five methionine supplementation levels (0, 1, 2, 4 and 6 g kg^?1), were hand‐fed twice daily to triplicate groups of Nile tilapia (initial body weight 8.95 ± 0.06 g) for 8 weeks. Weight gain (WG) and specific growth rate (SGR) increased significantly with increasing dietary methionine concentration at the same DE content (P < 0.001). At the same dietary methionine level, WG and SGR of fish fed high‐DE diets were significantly higher than that of fish fed low‐DE diets (P = 0.0001), although no interaction was found between dietary DE and methionine supplementation. Based on quadratic regression analysis between dietary methionine concentration and weight gain, optimal methionine requirement for maximum growth, expressed as g Met required kg^?1 diet (low‐ versus high‐DE diets), increased as diet DE concentration increased (7.34 versus 9.90 g kg^?1 diet, respectively; with cysteine 4.70 g kg^?1 diet). The results indicated that diet DE content affects methionine utilization and requirement in juvenile Nile tilapia, fish fed high‐DE diets required more methionine for maximum growth.     

Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.     

Molecular,biochemical and histopathological studies on the ameliorative effect of vitamin C on the renal and muscle tissues of Nile tilapia fish (Oreochromis niloticus) affected by the usage of engine oil

The present study was performed to evaluate the effect of vitamin C on the performance of Nile tilapia (Oreochromis niloticus) affected by the usage of engine oil. A total of 120 tilapia fish were randomly divided into four equal groups according to dietary treatments applied for 3 weeks: group I (control); group II was exposed to engine oil; group III was exposed to engine oil and supplemented with vitamin C: and group IV was fed only a diet supplemented with vitamin C. According to our results, the group exposed to engine oil (II) showed an induced elevation of kidney function parameters, an upregulation of CYP1A1 mRNA expression level, a reduction in antioxidant status (SOD, GSH) and severe abnormal histological and ultra‐structural changes in the head kidney, trunk kidney and muscle tissue. Supplementation of group III with vitamin C ameliorated the toxic effect induced by engine oil through improvement of kidney function parameters, elevation of the antioxidant status of tissue and improvement of the histological images of renal and muscle tissues. In conclusion, vitamin C has the ability to ameliorate engine oil oxidative stress toxicity in Nile tilapia.