Aurantiochytrium sp. meal as DHA source in Nile tilapia diet,part I: Growth performance and body composition

This study evaluated the inclusion of Aurantiochytrium sp. (ALL‐G‐RICH?), a source of docosahexaenoic acid (22:6 n‐3, DHA), in the diet of Nile tilapia and its effect on growth performance indexes and body composition. Fish (initial mean weight 8.35 ± 0.80 g) were fed different dietary inclusion levels of ALL‐G‐RICH?: 0.00, 0.05, 0.10, 0.20, and 0.40 g/kg and a control diet using cod liver oil (CLO), to provide DHA content comparable to the inclusion of 0.10 g/kg ALL‐G‐RICH?. Although there was no significant effect (p > 0.05) on weight gain, specific growth rate, feed conversion, protein retention rate, and proximal body composition, the inclusion of ALL‐G‐RICH? in the Nile tilapia diet influenced positively the fatty acid profile in the body, resulting in a high DHA concentration. CLO‐fed fish accumulated significantly more DHA compared to those fed 0.10 g/kg ALL‐G‐RICH? (p < 0.05). A digestibility trial was also performed for ALL‐G‐RICH with 65.86 g mean‐initial‐weight fish. The digestibility was high for DHA (96.10%); however, it was low for palmitic acid (70.81%). The results show that the inclusion of up to 0.40 g/kg ALL‐G‐RICH? can be used in Nile tilapia diets without impairing growth performance.     

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.     

Dietary ratios of n‐3/n‐6 fatty acids do not affect growth of Nile tilapia at optimal temperatures (28°C) nor at temperatures that simulate the onset of winter (22°C)

Nile tilapia (Oreochromis niloticus) juveniles were fed diets containing 13 g/kg total polyunsaturated fatty acids (PUFAs) at different n‐3/n‐6 dietary ratios (0.2, 0.5, 0.8, 1.3 and 2.9) for 56 days, at 28°C. Subsequently, fish were submitted to a winter‐onset simulation (22°C) for 33 days. PUFA n‐3/n‐6 dietary ratios did not affect fish growth at either temperature. At 28°C, tilapia body fat composition increased with decreasing dietary PUFA n‐3/n‐6. Winter‐onset simulation significantly changed feed intake. The lowest dietary n‐3/n‐6 ratio resulted in the highest feed intake. At both temperatures, body concentrations of α‐linolenic acid, docosahexaenoic acid, eicosatrienoic acid and docosapentaenoic acid decreased as dietary n‐3/n‐6 decreased. Body concentrations of eicosapentaenoic acid (EPA, 20:5 n‐3) increased with decreasing concentrations of dietary EPA. The n‐6 fatty acids with the highest concentrations in tilapia bodies were linoleic acid and arachidonic acid (ARA, 20:4 n‐6). At 28°C, SREBP1 gene expression was upregulated in tilapia fed the lowest n‐3/n‐6 diet compared to tilapia fed the highest n‐3/n‐6 ratio diet. Our results demonstrate that a dietary PUFA of 13 g/kg, regardless of the n‐3/n‐6 ratio, can promote weight gains of 2.65 g/fish per day at 28°C and 2.35 g/fish per day at 22°C.     

Availability of minerals in rendered meat and bone meal for Nile tilapia: Preliminary observations

A digestibility trial was conducted with 180 Nile tilapia (32.65 ± 4.52 g) fed a reference diet containing 350 g/kg crude protein and 4,500 kcal/kg gross energy. Five different meat and bone meals (MBMs) were evaluated. The test diets compound by 70% of reference diet and 30% of each MBM were elaborated and classified according to ash content: MBM‐430, MBM‐400, MBM‐370, MBM‐340 and MBM‐310. The apparent availability coefficient (AAC) of minerals and the apparent digestibility coefficient (ADC) of dry matter (DM) were subjected to one‐way analysis of variance (p < .05). Differences among ADC and AAC were determined by Tukey's multiple range test (p < .05). When appropriate, a polynomial regression analysis was performed. Linear correlation analyses between MBM chemical composition and the AAC and ADC were performed (p < .01). Higher values of ADC_DM, AAC_Ca and AAC_P were observed in MBM‐310, while low values were observed in MBM‐430. Quadratic effects of ash, P and Ca contents on AAC_Ca, AAC_P and ADC_DM were observed. The AAC_Ca, AAC_P and ADC_DM indicated a negative correlation with ash, P and Ca contents. MBM‐310 exhibited the highest mineral AAC, which could be used as a mineral source for 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.     

Yeast extract on growth,nutrient utilization and haemato‐immunological responses of Nile tilapia

Dietary supplementation (0, 10, 20, 40, 60, 80 g kg^?1) of a nucleotide source (NuPro^®) was evaluated on growth, nutrient utilization, haemato‐immunological responses and resistance to experimental infection of Nile tilapia. Fish (2.63 ± 0.63 g) were fed experimental diets to satiation three times a day for 75 days. Feed intake increased linearly with increasing levels of NuPro^® promoting higher weight gain (P < 0.05). Survival, feed conversion and protein retention were not affected. Following the growth trial, fish were challenged with Aeromonas hydrophila by intraperitoneal injection. Survival and innate immune responses (phagocytosis and lysozyme activity) were not significantly affected. However, among the haematological variables, thrombocyte, leucocyte and monocyte counts increased linearly (P < 0.05). The indirect measurement of digestibility in juvenile Nile tilapia (122.32 ± 11 g) indicated a linear decrease in protein (1.41%), dry matter (5.72%) and energy (4.10%) digestibility coefficients as dietary supplementation increased (P < 0.05). NuPro^® showed to be a beneficial additive to be supplemented in diets for juvenile Nile tilapia, providing 32.8% increase in feed intake and 28.8% in weight gain, as well as in thrombocyte, leucocyte and monocyte counts in blood, possibly indicating a more efficient defence response in the highest dietary supplementation levels tested.     

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 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.     

Effects of biofloc technology on reproduction and ovarian recrudescence in Nile tilapia

The objectives of this study were to evaluate the reproductive cycle, morphological changes of ovary and mobilization of energy reserves in Nile tilapia reared with biofloc technology (BFT). In general, the growth and reproductive performance were highly similar between BFT and Control system (clear water). Difference between the systems was found in the hepatosomatic index (using mixed‐effects models), which suggested that BFT can alter the energy mobilization in the post‐spawning period. The absolute and relative fecundity, fertilization rate, number of larvae produced per female, gonadosomatic index, proportion of oogenesis cells, number of post‐ovulatory and atretic follicles were similar between the two systems. We also did not detect a reduction in the reproductive cycle length in Nile tilapia reared in BFT. Because there was no evidence of the negative effects of BFT on Nile tilapia reproduction, we concluded that BFT might be used for breeder stocking of this species.     

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.     

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%.     

Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia

We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 10⁴ cfu mL^?1 (Treatment 1) and 1.0 ×10⁵ cfu mL^?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 10⁷ cfu g^?1 (Treatment 1) and 1.0 × 10⁸ cfu g^?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.     

Orange peel fragment improves antioxidant capacity and haematological profile of Nile tilapia subjected to heat/dissolved oxygen‐induced stress

This study evaluated the ability of orange peel fragment (OPF) to act as a functional feedstuff, influencing growth, haematological profile, and antioxidant enzyme activity of Nile tilapia subjected heat/dissolved oxygen‐induced stress (HDOIS). A group of 440 male Nile tilapia (31.7 g ± 0.34) was randomly distributed in 40 250‐L aquaria (11 fish/tank) and fed five practical diets with graded levels of OPF at 0%, 0.2%, 0.4%, 0.6%, and 0.8% for 70 days. The diets were formulated to contain 30% crude protein and 18 MJ/kg crude energy. After the feeding period, growth performance was evaluated and six fish per treatment were sampled for haematological profile and antioxidant enzyme activity, before and after HDOIS. Then, fish were subjected to HDOIS (32°C/2.3 mg/L dissolved oxygen) for three days and the same haematological profile and antioxidant enzyme activity were determined. There was no effect of OPF on the haematological profile, either before or after HDOIS. The polynomial regression model was used to express the relationship between antioxidant enzymes activity and OPF supplementation level. The maximum activity of superoxide dismutase, catalase, and glutathione peroxidase was reached at 0.66%, 0.63%, and 0.68% of OPF respectively. Results of the present study suggest that a dietary supplementation level of 0.63%–0.68% of orange peel fragment was appropriate to maintain Nile tilapia haematological profile and improve its antioxidant capacity under HDOIS.     

Dietary lipid sources affect the performance of Nile tilapia at optimal and cold,suboptimal temperatures

Five sources of dietary fatty acids (fish, linseed, sunflower, olive and coconut oils) were evaluated in juvenile Nile tilapia in two trials: at optimal (28°C) and suboptimal (22°C) temperatures lasting 9 and 12 weeks, respectively. At 28°C, there was no clear effect of dietary source on fish growth, but at 22°C, the highest daily weight gain occurred in fish fed sunflower, linseed and fish oil. Feed efficiency and apparent net protein utilization increased as the amount of unsaturated fatty acids, especially n‐3 polyunsaturated fatty acids (PUFA), in the diet increased. Coconut oil, which is rich in saturated fatty acids (SFA), led to the worst growth results, especially at 22°C, with the lowest weight gain, feed intake and feed utilization by tilapia. The body fatty acid profile, in % of total fatty acids, was dependent on diet composition. However, for all treatments, PUFA body content increased with the decrease in temperature, but SFA and monounsaturated fatty acids remained the primary contributors to the body profile. Either fish oil or vegetable oil may be used as sources of dietary fatty acids for Nile tilapia, but at suboptimal temperatures, a dietary source containing more PUFA and less SFA improves performance.     

Earthworm,Eisenia fetida,bedding meal as potential cheap fishmeal replacement ingredient for semi‐intensive farming of Nile Tilapia,Oreochromis niloticus

The study amalgamated earthworm and agro‐industrial wastes through vermicomposting and then evaluated the potential of the bedding (mixture of Eisenia fetida and vermicompost) to replace fishmeal in semi‐intensive farming of Oreochromis niloticus. The bedding was used to substitute fishmeal at inclusion rates of 100, 60, 30 and 0% (D100, D60, D30 and control D0). In triplicates, the four homogeneous diets were fed to quadruplicate groups of 30 g O. niloticus for 112 days. There was no significant difference (p > .05) in mortalities, average length gain and FCR among all tests. Nevertheless, diet D0 had significantly (p < .05) superior amino acid profile, low fibre content and fish carcass crude protein (63.2 ± 0.72% dry matter). Subsequently, D30 and D0 produced fish with significantly higher (p < .05) mean weight gain (256.03 ± 0.4 g) and biomass (369,136 g) respectively. On to the contrary, diet D100 had significantly higher (p < .05) crude lipids content (9.4 ± 0.6% dry matter), economic returns and profit index than the control diet due to the comparatively low cost of producing the earthworm bedding. This simple biotechnology can commercially be upscaled to sustainably produce cheap and nutritious fish feed capable of increasing yields and maximizing profits.     

Dietary addition of the essential oil from Lippia alba to Nile tilapia and its effect after inoculation with Aeromonas spp.

The objective of this study was to evaluate the effect of dietary addition of the essential oil from Lippia alba (EOLA) on growth performance, biochemical and haematological variables and survival after Aeromonas spp. infection of Nile tilapia juveniles (Oreochromis niloticus). Five diets were evaluated with increasing levels of EOLA (0.0 control; 0.25; 0.50; 1.0; and 2.0 ml/kg diet). After 45 days of feeding, the fish were infected with Aeromonas spp. followed by a 14‐day period of observation. There was no mortality during growth assessment. The addition of 2.0 ml EOLA/kg diet improved feed conversion ratio and condition factor and increased lysozyme activity and haematocrit (Hct), and decreased plasma globulin levels compared to the control group. The survival of fish infected with Aeromonas spp. was higher in those fed with 2.0 ml EOLA/kg diet than in fish fed with 0.0–0.5 ml EOLA/kg diet. We concluded that the inclusion of 2.0 ml EOLA/kg diet is recommended for Nile tilapia juveniles, because it improved feed conversion, Hct and immunological activity, did not provoke metabolic changes, and increased survival after Aeromonas spp. infection.     

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.     

The effect of plant protein‐based diets on apparent nutrient digestibility,growth response,egesta quantity,postprandial ammonia excretion rate and serum quality of Nile tilapia

The study assessed the effect of oilseed meal mixtures on the biological value, faecal matter output, ammonia excretion rates and serum biochemistry of Nile tilapia over 63 days. The experimental diets (PPB 1, PPB 2 and PPB 3) were formulated using four selected oilseed meals that were mixed in different protein proportions to contribute 80% of total dietary crude protein. In each diet, either one or two of the oilseed meals were the dominant protein contributor. A commercial tilapia feed was used as the control diet (CTRL). Sex‐reversed Nile tilapia fingerlings (35 g) were stocked at 20 fish per tank in a recirculation system and fed at 3% body weight of their respective diets. At the end of the study, ADCs of dry matter, crude lipid and ash were significantly (p < .05) lower in all the plant‐based diets compared with the control diet. Fish fed the control diet had significantly higher weight gain and SGR compared with the plant‐based diets. Egesta output was 127% higher in the plant‐based diets compared with the control diet. All dietary treatments exhibited a similar trend in changes in ammonia nitrogen. Nevertheless, serum metabolites levels indicated no significant differences among treatments. Although the PPBs did not affect ADC of protein and serum profile, fish growth was reduced while faecal output increased.     

Aurantiochytrium sp. meal can replace fish oil in practical diets for the juvenile Pacific white shrimp

This study evaluated the use of microalgae (Aurantiochytrium sp.) meal as a substitute for fish oil in the diet of juvenile Pacific white shrimp (Litopenaeus vannamei) reared in a clear‐water system. Dietary treatments at five replacement levels (0%, 25%, 50%, 75%, 100%) were performed in triplicate. After 46 days, only a slight difference in shrimp final weight was observed among treatments (0.61 g). An increase in final weight was observed with replacement of up to 50% fish oil for microalgae meal, while the optimal percentage of replacement estimated was 44.7%. Feed conversion rate (FCR) of animals fed a diet of Aurantiochytrium sp.meal to replace up to 50% fish oil decreased, and the optimal percentage of replacement estimated was 49.3%. The fatty acids profile in shrimp muscle tissue demonstrated an increase in docosahexaenoic acid (DHA) from 10.03% to 14.28% with increased replacement of fish oil by microalgae meal in the diet. Therefore, the partial replacement of fish oil by microalgae meal resulted in improved shrimp growth and FCR, and total replacement of dietary fish oil had no negative effects on these parameters. In addition, inclusion of microalgae meal raises the level of DHA in shrimp muscle.