Stable isotope analysis to quantify contributions of supplementary feed in Nile Tilapia Oreochromis niloticus (GIFT strain) aquaculture

Nile tilapia (Oreochromis niloticus) is currently one of the most farmed freshwater fish and contributes significantly to total global aquaculture production. The genetically improved strain of O. niloticus (GIFT) was introduced to Papua New Guinea (PNG) in 1999 to improve food and income security. The high cost and low availability of commercial fish feed hinder the growth of GIFT farming in PNG. Stable carbon and nitrogen isotopes were used to determine the role of supplementary and natural food sources in the diet of GIFT in pond‐based aquaculture. Two treatments were used: treatment 1 was daily feeding, and treatment 2 was weekly feeding, each with three replicates. Isotopic analysis of muscle tissue and all potential food sources showed that pellet feed contributed 7% to the growth of GIFT in daily‐fed ponds and 33% in the weekly‐fed ponds. Highly enriched δ¹⁵N values for chicken manure, compared to depleted values for GIFT and other natural food sources in both treatments, clearly indicate insignificant contributions of this input to production. After 90 days of cultivation, the average final body weight of GIFT receiving daily feed inputs was 134 g (average 19 cm), while for weekly‐fed it was 92 g (17 cm). The feed conversion ratio (FCR) was poor (6.4:1) in the daily‐fed GIFT ponds compared to a better, and preferable, FCR (1:1) in the weekly‐fed ponds. The findings of this study show that pelleted feed was not the major contributor to the growth of GIFT. Genetically improved farmed tilapia aquaculture should focus on enhancing natural food availability for fish production.     

Evaluation of hydrolysed rubber seed meal as a dietary protein source for Nile tilapia Oreochromis niloticus L.

This study evaluated the use of hydrolysed rubber seed meal as a locally grown alternative feed ingredient to reduce the use of fishmeal within formulated feeds for tilapia. Five experimental diets were prepared by formulating different inclusion levels of hydrolysed rubber seed meal (0%, 25%, 50%, 75% and 100% of plant‐based protein contribution). Dried and defatted rubber seed meal was hydrolysed by adding rumen digesta liquid to the ingredient. For digestibility measurement, chromic oxide was added to each diet as an inert marker. The digestibility trial was conducted over a 14‐day period by collecting faecal matter from the rearing tanks. In addition, a growth trial was conducted over a period of 50 days using four replicates per dietary treatment. In both experiments, all fish were fed three times daily to apparent satiation. At the end of growth trial, the inclusion level of hydrolysed rubber seed meal up to 50% of plant‐based protein contribution did not give any significant difference compared to the control diet in terms of feed efficiency, specific growth rate and protein retention parameters. In the digestibility study, fish fed diets containing hydrolysed rubber seed meal up to a 50% of plant‐based protein inclusion level did not give any significant difference compared to the control diet in terms of protein and dry matter apparent digestibility. Based on the result of this growth and digestibility study, up to a 50% of plant‐based protein contribution of hydrolysed rubber seed meal can be used within formulated tilapia feeds without any adverse effect.     

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.     

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.     

Utilization of Caridina nilotica (Roux) meal as a protein ingredient in feeds for Nile tilapia (Oreochromis niloticus)

The effects of replacing fish meal with Caridina nilotica as a protein ingredient on growth performance, nutrient utilization, carcass, proximate composition and economic benefits in Nile tilapia (Oreochromis niloticus) culture was evaluated. Replacement of the FM with C. nilotica was done at 25%, 50%, 75% and 100% (D25, D50, D75 and D100) and the substitution effects was compared with the control diet (D0, 0% C. nilotica). After 140 days of culture, the best growth performance, nutrient utilization and economic benefits occurred in fish groups fed diets with 25% C. nilotica inclusion. However, growth performance in fish fed diets D50 and D75 were comparable with the control (P > 0.05). At 100% substitution level of FM with C. nilotica, the growth performance and fish survival was lower than control. Protein and lipid contents in the fish and their digestibilities were highest in diet D25 and decreased with increasing levels of substitution of FM with C. nilotica. This study demonstrate that utilization of local protein sources (C. nilotica) can be effectively used to replace up to 75% of FM in the diets without compromising growth performance, survival, nutrient utilization and economic benefits in O. niloticus culture.     

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%.     

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.     

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.     

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.     

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.     

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.     

Physiological responses and HSP70 mRNA expression of GIFT strain of Nile tilapia (Oreochromis niloticus) under cold stress

An investigation on the physiological responses and HSP70 expression under cold stress was conducted on Genetically Improved Farmed Tilapia (GIFT) strain of Nile tilapia (Oreochromis niloticus). Tilapia were acclimated at 28°C as control and then the water temperature was reduced from 28°C to 15°C at a rate of 1°C h^?1, and serum biochemical indices, antioxidant enzymes and expression of HSP70 mRNA were analysed on days 0, 0.5, 1, 2, 3 and 5 after exposure to 15°C. The concentration of serum K, Na, Cl and Ca concentration showed instability during cold stress. Glucose rapidly increased on day 0 followed by a declining trend. Triglyceride, cholesterol, low density lipoprotein‐cholesterol signifi‐cantly decreased and high density lipoprotein‐cholesterol showed instability during cold stress. Aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase activities were prominently elevated under cold stress. Superoxide dismutase and catalase activities showed a remarkable rise on day 0.5 under cold stress, and gradually decreased thereafter. HSP70 mRNA level significantly increased both in liver and muscle under cold stress, especially on days 0.5 and 1. These results suggest that cold influences several physiological responses of tilapia, and the cold resistance and cold tolerance of tilapia will benefit from the enhancement of antioxidant enzymes activities and upregulated HSP70 mRNA expression.     

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.     

Evaluation of nutritive value of fermented de‐oiled physic nut,Jatropha curcas,seed meal for Nile tilapia Oreochromis niloticus fingerlings

Jatropha curcas seed meal was fermented with Bacillus licheniformis (LFJSM) and Bacillus pumilus (PFJSM) separately using the solid‐state fermentation. After fermentation, the crude protein and the total hydrolysed amino acid contents were increased in LFJSM and PFJSM, while fibre content, phytic acid, trypsin inhibitor and saponins were decreased. An 84‐day feeding experiment was designed for the nutritional evaluation of LFJSM and PFJSM in Nile tilapia. Seven isonitrogenous (305.47 g kg^?1 crude protein) and isocaloric (18.21 MJ kg^?1 gross energy) diets were formulated by replacing 0% (control diet), 25% (LFJSM‐25), 50% (LFJSM‐50), 75% (LFJSM‐75) and 25% (PFJSM‐25), 50% (PFJSM‐50) and 75% (PFJSM‐75) of protein from fish meal with LFJSM and PFJSM. The higher weight gain and the best feed conversion ratio were found in fish fed control diet, PFJSM‐25 and PFJSM‐50, which were insignificantly different. Apparent digestibility coefficient values of dry matter, crude protein, lipid and digestible energy reached a plateau in fish fed PFJSM‐25 and PFJSM‐50. No significant differences were found in haematocrit, haemoglobin, serum alternative complement pathway components, alanine aminotransferase and aspartate aminotransferase and triglycerides between the control diet, LFJSM‐25, LFJSM‐50, PFJSM‐25 and PFJSM‐50 treatment groups. Therefore, these findings suggest that up 50% of fish meal can be replaced by PFJSM in Nile tilapia diets.     

Performance of mono‐sex tilapia (Oreochromis niloticus) in rice field with different ditch size

The effects of ditch size on growth and production of mono‐sex tilapia, Oreochromis niloticus in rain fed concurrent rice–fish system were technically and economically evaluated for a period of 4 months. Three different ditch sizes were tried: 5%, 10% and 15% of the total cultivable rice field. The fish fingerlings stocking density (5000 ha^?1) and management practices were similar in all plots. The water temperature, dissolve oxygen, pH, Secchi disc visibility, ortho‐phosphate, ammonia nitrogen and alkalinity were found to be in the range of 31.5–31.9°C, 5.10–5.27 mg L^?1, 7.56–7.62, 34.81–35.41 cm, 0.88–0.96 mg L^?1, 0.014–0.021 mg L^?1 and 66.23–69.76 mg L^?1, respectively, and all the values were found suitable for tilapia fish farming. Sediment organic matter, available nitrogen and phosphorous were increased significantly (P < 0.05) in all plots after cultivating the fish. Larger ditch size (15% of the total land area) resulted in significantly (P < 0.05) higher specific growth rate, survival and fish production compared with the smaller (5%) and medium (10%) ditch sizes. However, significantly (P < 0.05) higher net income and benefit–cost ratio were obtained in medium ditch size with the values of 2.6% and 3.6% higher than the smaller and larger ditch sizes respectively. It is concluded that rice–fish farming in a rain fed ecosystem of Bangladesh with medium ditch size and stocking density of 5000 ha^?1 mono‐sex tilapia can achieve better economic return.     

A new approach to fishery waste revalorization to enhance Nile tilapia (Oreochromis niloticus) digestion process

The main goal of this research was to analyse in vitro compatibility of Nile tilapia (Oreochromis niloticus) digestive proteinases and enzymes recovered from species comprising fishery waste as Pleoticus muelleri, Artemesia longinaris and Patagonotothen ramsayi. It evaluated the use of exogenous enzymes as feed supplements to increase digestion efficiency in tilapia fingerlings and juveniles (3.5 ± 0.11 g and 11.6 ± 1.5 g, respectively). We successfully have obtained proteinases from fishery waste as source of exogenous enzymes. P. muelleri and A. longinaris enzymes had more activity of acid and alkaline proteinases than P. ramsayi. SDS‐PAGE gels demonstrated that Nile tilapia digestive proteinases keep their activity when combined with each exogenous proteinase. Exogenous enzymes varied in their ability to enhance hydrolysis of different feed ingredients. P. muelleri by‐products are the best candidates to be employed as feed supplements for tilapia juveniles. Enzymes from this by‐product did not affect the activity and integrity of fish digestive enzymes, improved the hydrolysis of different protein sources (fish meal, squid meal, shrimp meal and soybean meal), and maintained its activity after being exposed to high temperatures and acid pHs. Our findings are applicable to other places where O. niloticus is raised utilizing local fishery waste, and also to different cultured species.     

Methionine and lysine requirements for maintenance and efficiency of utilization for growth of two sizes of tilapia (Oreochromis niloticus)

Different ration levels were used to determine the digestible methionine (DMet) and lysine (DLys) maintenance requirements and the utilization efficiencies for gain above maintenance for two different sizes of tilapia (20.7 and 165 g), by feeding a soybean meal–based diet. Protein gain and amino acid (AA) gain (e.g. methionine, Met; lysine, Lys; R² = 0.98) were best‐fit linear functions of DMet and DLys intake in both fish size classes. Slopes of these regression lines showed that the DMet utilization efficiencies for growth were 0.76 and 0.55 for juvenile and adult fish, respectively. The DMet maintenance requirements were 3.12 and 16.5 mg BW(kg)^−0.7 day⁻¹ for juvenile and adult fish, respectively. The DLys utilization efficiencies for gain were 0.72 and 0.52, whereas the DLys maintenance requirements were 16.9 and 68.8 mg BW (kg)^−0.7 day⁻¹, for juvenile and adult fish, respectively. These results suggested that there was an obvious difference in the maintenance requirements and utilization efficiencies for gain above maintenance for DMet and DLys in two different sizes of tilapia. The AA maintenance needs increased as fish increased in size, being greater in adult fish than in juvenile; however, the AA utilization efficiencies for gain above maintenance decreased with the increment of fish size.     

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.