The effect of plant‐based diet and suboptimal environmental conditions on digestive function and diet‐induced enteropathy in rainbow trout (Oncorhynchus mykiss)

This experiment investigated intestinal enteropathy and digestive function of rainbow trout challenged with soybean meal‐based diet (SBM) at optimal or suboptimal environments created by normal or reduced water flow, respectively. Oxygen level remained above 7 mg L^‐1 for optimal environment and between 4 and 5 mg L^‐1 for suboptimal environment. Triplicate groups of fish (mean body weight 74 g) were fed fishmeal‐based diet (FM) or SBM at optimal environment in period 1 (28 days). In period 2 (42 days), fish were subjected to a change from FM to SBM or remained on the same diet as used in period 1. The fish were also exposed to change from optimal to suboptimal environment or remained under optimal conditions. The fish subjected to change from FM to SBM, regardless of their environment, showed similar degree of enteropathy from day 14. Lipid and starch digestibility was lower in SBM‐fed fish at suboptimal environment compared to fish fed the same diet at optimal environment. Crude protein digestibility, however, was highest in SBM‐fed fish at suboptimal environment throughout period 2. In conclusion, in SBM‐fed rainbow trout, exposure to suboptimal environment did not change the degree of enteropathy; however, lipid and starch digestibility were further reduced.     

Phytase and sodium diformate supplementation in a plant‐based diet improves protein and mineral utilization in rainbow trout (Oncorhynchus mykiss)

A basal isonitrogenous and isoenergetic plant‐based diet (Control) was supplemented with either 10 g kg^?1 sodium diformate (NaDF), 4000 FTU kg^?1 phytase (Phy) or a combination of both additives (NaDF + Phy). Three hundred juvenile rainbow trout with an average weight of 120 g were randomly distributed into 12 fibreglass tanks (300 L). After 65 days of trial, fish fed diets containing phytase, NaDF or the combination of both additives showed a higher growth rate (P < 0.05) compared to fish fed Control diet. NaDF increased feed intake (P = 0.032), while phytase inclusion resulted in a better feed conversion ratio (P < 0.0001) and a higher N retention efficiency (P = 0.02) compared with the Control. Apparent digestibility of P, Ca, Mg and Zn was improved by the use of phytase (P < 0.005) as well as P, Ca and Mg retention efficiency in fish (P < 0.0001). Using 4000 FTU kg^?1 phytase in plant‐based diets resulted in a 13% and 50% reduction in N and P loadings, respectively. The use of NaDF in combination with phytase in a plant‐based diet for rainbow trout resulted in a higher weight gain than that when NaDF was used alone.     

Digestibility, growth and nutrient retention in rainbow trout (Oncorhynchus mykiss) fed diets extruded at two different temperatures

Digestibility and utilization of a fishmeal‐based diet extruded at 103 or 137 °C were examined. Each of the diets was fed to 0.5‐kg rainbow trout in nine tanks supplied with freshwater. Specific growth rate was significantly (P < 0.05) elevated for trout fed the feed extruded at 137 °C. The apparent digestibilities of protein and energy, feed conversion ratio, retentions of nitrogen and energy were not significantly affected by extrusion temperature. Digestibility of ash was significantly reduced in the diets produced at 137 °C.     

Digestibility, growth and excretion response of rainbow trout (Oncorhynchus mykiss Walbaum) to feeds of different ingredient particle sizes

The effects of feed ingredient particle size on apparent digestibility, growth and excretion responses of rainbow trout were investigated in pilot production systems. Particle size of the feed ingredients (ground using a hammer mill with 0.6 and 3.0 mm screen sizes) had no significant effect (P > 0.05, d.f. = 1/6) on the apparent digestibility of dry matter, phosphorus and crude protein in cold‐extruded diets. Feed ingredient particle size had an impact on the feed conversion ratio over an 85‐day feeding trial when key ingredients (78.65% of diet) were ground using a pulverizer and a hammer mill with 3.0 mm screen size and heat extruded. No significant differences (P > 0.05, d.f. = 1/12) were observed in the excretion ratios of TAN (total ammonia nitrogen), TKN (total Kjeldahl nitrogen), TP (total phosphorus), BOD₅ (5‐day biochemical oxygen demand) and TSS (total suspended solids) among three feed types, which included a wide ingredient particle size distribution.     

Supplementing rainbow trout (Oncorhynchus mykiss) broodstock diets with choline and methionine improves growth in offspring

Nutritional programming represents the mechanism through which broodstock feeding strategies have lifelong effects on the offspring generation. The objective of this study was to determine whether supplementing rainbow trout, Oncorhynchus mykiss broodstock diets with choline and methionine affects offspring growth performance. Experimental diets were produced by top-dressing a commercial broodstock diet with (a) 12,000 ppm methionine, (b) 7,400 ppm choline, (c) 12,000 ppm methionine and 7,400 ppm choline, or (d) water (control). Six rainbow trout families were fed experimental diets beginning 18 months post-hatch through spawning; the offspring consumed a commercial diet. The broodstock diet did not affect maternal body weight, body condition, egg size, or egg yield (p > .05). Eggs from choline-treated broodstock contained 10% more choline than control eggs (p < .05). Offspring from choline-treated broodstock were smaller than controls at 146 days post-hatch (dph) (p < .05). At the final harvest (439 dph), offspring from the control broodstock weighed 12–18% less than offspring from broodstock consuming the supplemented diets (p < .05). However, there was a significant diet-by-family interaction on offspring growth (p < .05); supplemented diets improved performance in three of the six broodstock families. These findings indicate that supplementing broodstock diets with methionine and choline causes a nutritional programming effect that benefits growth in offspring.     

Using Near Infrared Reflectance Spectroscopy (NIRS) to predict the protein and energy digestibility of lupin kernel meals when fed to rainbow trout,Oncorhynchus mykiss

This study examined the potential of using near infrared spectroscopy (NIRS) to predict the nutrient composition, energy density and the digestible protein and digestible energy values of lupin kernel meals when fed to rainbow trout. A series of 136 lupin kernel meals were assessed for their protein and energy digestibilities using the diet‐substitution approach in a series of 10 experiments over a 6‐year period from 2002 to 2008. Two reference diets were also included in each experiment. Minimal variance in the digestibility parameters of both reference diets was observed among the experiments ensuring that there was a high degree of robustness in the across‐experiment evaluations. The same lupin kernel meal samples were also scanned using a diode array near infrared spectrophotometer (DA‐NIRS). The spectra obtained by the DA‐NIRS were chemometrically calibrated against both the chemical composition and the digestible value data using multivariate analysis software. The cross validation tests used in this study provide a valid indication of the potential to predict the nutrient composition, energy value and digestible protein and energy values of the lupin kernel meals as used in diets for rainbow trout. That the standard errors of cross validation (SECV) of the parameters investigated were generally commensurate with the cross trial variation seen in the reference sample indicating robust calibrations for the two target parameters of digestible protein and digestible energy. Therefore this study demonstrates that within one raw material type that not only does significant variability in the digestible value of the raw materials exist, but that it is possible to use NIRS technology to provide rapid estimates of the digestible value of those raw materials in near real‐time.     

An in vitro method to determine phosphorus digestibility of rainbow trout Oncorhynchus mykiss (Walbaum) feed ingredients

An in vitro method was developed to assess the digestibility of phosphorus in 12 plant and animal feed ingredients for rainbow trout Oncorhynchus mykiss (Walbaum). The method simulates the gastrointestinal tract of the rainbow trout with regard to pH and gastrointestinal enzymes. Phosphorus solubility was measured after acid digestion (pH 3) with and without gastric enzymes, after alkaline digestion (pH 9) with and without intestinal enzymes, and after a two-step process involving acid and alkaline digestion. Commercially available digestive enzymes from mammals were compared with digestive enzymes from rainbow trout. Correlating in vitro digestibility with in vivo digestibility showed that acid digestion with both commercial enzymes ( r ²=0.98, P  < 0.05) and trout enzymes ( r ²=0.94, P  < 0.05) predicted the in vivo digestibility of animal feed ingredients. Alkaline digestion with both enzyme systems (commercial r ²=0.79; trout r ²=0.74, P  < 0.05) or without ( r ²=0.82, P  < 0.05) enzymes predicted the in vivo digestibility of ingredients from animal byproducts but not those from plant products. The in vitro digestibility with two enzyme steps (acid and alkaline) predicted in vivo digestibility of plant and animal ingredients ( r ²=0.79 for commercial enzymes and r ²=0.74 for trout enzymes) better than did one-step acid or alkaline digestion.     

Aqueous fractionation improves the nutritional value of wheat distillers grains for rainbow trout (Oncorhynchus mykiss)

Fractionation of wheat wet distillers grains (WWDG) was performed to reduce non‐starch polysaccharides (NSP) and increase protein content. WWDG was collected from two ethanol plants, and protein was extracted by the addition of water, autoclaving to precipitate protein and drying in a forced air oven at 55 °C to create WWDG–protein concentrate (WWDG–PC). This process increased crude protein (CP) and decreased NSP in the WWDG–PC (P < 0.05). Dry matter and energy digestibility of WWDG–PC in rainbow trout were increased by fractionation (P < 0.05). A 56‐day growth trial was performed to determine the effect of feeding WWDG–PC on growth performance of rainbow trout. Rainbow trout (n = 18/tank; body weight = 231 g; 3 tanks/treatment) were fed diets containing 0, 75, 150, 225 or 300 g kg^?1 WWDG–PC. All diets were formulated to contain 386 g kg^?1 digestible CP and 17.6 MJ kg^?1 digestible energy. There was no significant linear or quadratic relationship between inclusion rate and growth, feed intake or feed conversion, indicating that at inclusion levels of up to 300 g kg^?1 of WDDG–PC does not reduce the growth performance of rainbow trout.     

Effect of microbial phytase on apparent nutrient digestibility of barley, canola meal, wheat and wheat middlings, measured in vivo using rainbow trout (Oncorhynchus mykiss)

Microbial phytase (Natuphos 5000G) was supplemented into barley, canola meal, wheat and wheat middlings at 500 FTU kg^–1 diet to test the effectiveness of the phytase on digestibility of crude protein (CP), gross energy (GE) and minerals for rainbow trout. A total of 180 rainbow trout (initial mean body weight 223.8 ± 17.9 g) were stocked into ten 40-L fibreglass digestibility tanks with 18 fish per tank. Duplicate tanks were assigned to a reference diet and a single tank was assigned randomly to each of the eight diets made from these four ingredients supplemented with and without phytase. The collection of faeces lasted for 2 weeks. Faeces collected in each week represented a replicate and they were analysed separately. The mean apparent digestibility coefficients (average of four ingredients) in phytase supplemented and nonsupplemented ingredients were: CP, 97.3% and 96.5%; GE, 62.8% and 53.1%; calcium (Ca), 38.3% and –5.3%; magnesium (Mg), 72.4% and 50.3%; manganese (Mn), 31.0% and –4.2%; total-phosphorus (total-P), 71.1% and 47.6%; phytate-phosphorus (phytate-P), 87.5% and 22.3%; copper (Cu), 38.3% and 26.4%; potassium (K), 97.1% and 97.0%; sulphur (S), 90.2% and 88.6%; zinc (Zn), 12.3% and –16.6%. A two-tailed t -test indicated that phytase supplementation significantly ( P  < 0.05) improved the digestibility of Ca, Mg, Mn, total-P, phytate-P, and GE. The efficacy of phytase on nutrient digestibility also depended on the type of ingredient as measured in rainbow trout.     

Performance and immunological responses of rainbow trout (Oncorhynchus mykiss) fed bioprocessed plant‐based proteins

Marine‐derived fish meal (FM) is a traditional component of commercial aquaculture feeds for many farmed fish species. Modern bioprocessing technologies have been developed to produce high‐protein plant‐based ingredients for aquafeeds to further reduce FM and other conventional animal protein sources. A 90‐day feeding trial using juvenile rainbow trout (Oncorhynchus mykiss) (27.9 ± 1.4 g; mean ± SE) was completed to evaluate growth, feed efficiency, general health and immunological responses to diets containing experimental plant‐based protein sources and reduced FM. Trout were fed one of four dietary treatments: experimental bioprocessed soy protein concentrate (BSPC), commercially available enzymatically hydrolysed soy protein concentrate (CSPC), experimental bioprocessed barley protein concentrate (BBPC) and a FM control (FMC). At trial termination, there were no significant differences in relative growth (RG) or specific growth rate (SGR) between fish fed either FMC or BSPC (p > .05). Fish fed either CSPC or BBPC resulted in significantly lower RG and SGR than the FMC (p < .05). Significant respiratory burst analysis differences were observed among treatments at day 90 (p < .01), for fish fed either FMC or CSPC diets. This assessment of bioprocessed plant‐based protein ingredients facilitates the characterization and incorporation of bioprocessed soy and barley protein as the industry continues to evaluate FM replacements in rainbow trout feeds.     

Replacement of fishmeal by canola protein concentrate in diets fed to rainbow trout (Oncorhynchus mykiss)

A dephytinized protein concentrate prepared from canola seed (CPC) was assessed for nutrient digestibility and performance in rainbow trout (Oncorhynchus mykiss). The apparent digestibility coefficients of CPC were: dry matter, 817 g kg^?1; crude protein, 899 g kg^?1; gross energy, 861 g kg^?1; arginine, 945 g kg^?1; lysine, 935 g kg^?1; methionine, 954 g kg^?1; threonine, 893 g kg^?1. A 9‐week performance trial assessed 7 diets. Fishmeal provided 940 g kg^?1 of the protein in the control diet. Test diets consisted of CPC or water‐washed CPC replacing 500 and 750 g kg^?1 of fishmeal protein; and CPC plus an attractant replacing 500 and 750 g kg^?1 of fishmeal protein. No significant differences in performance were observed (P > 0.05). A subsequent 9‐week performance trial evaluated the effect of adding CPC into compound diets containing fishmeal/soybean meal/corn gluten meal. Five diets were prepared: fishmeal provided 670 g kg^?1 of the protein in the control diet, in the remaining diets CPC was incorporated into commercial‐like trout diets at 100, 200 and 300 g kg^?1 replacement of fishmeal protein, the fifth diet included an attractant in the 300 g kg^?1 replacement diet. No significant differences in performance were obtained (P > 0.05). These studies show that dephytinized canola protein concentrate has potential to replace substantial levels of fishmeal in diets for carnivorous fish without compromising performance.     

Effect of enzyme supplementation to dehulled lupin-based diets on growth, feed efficiency, nutrient digestibility and carcass composition of rainbow trout, Oncorhynchus mykiss (Walbaum)

High inclusion levels of dehulled lupin (Lupinus angustifolius) in salmonid diets significantly decrease growth rates. This may be caused by the high concentrations of non‐starch polysaccharides including oligosaccharide (OS) in lupin. The antinutritive effects of OS have not yet been fully investigated in fish. The objective of this study was to determine the effect of enzyme supplementation of dehulled lupin‐based diets on the fish performance. There were two control diets: a fish meal‐based diet with no plant protein (FM) and a diet that contained 50% dehulled lupin (L). Four experimental diets based on diet L and containing four different exogenous enzyme supplements were used: diet L(E) (Energex^™); diet L(B) (Bio‐Feed^™ Pro); diet L(α) (Alpha galactosidase^™); and diet L(Mix), which contained all the enzymes. Fish were randomly stocked into tanks in duplicate groups of 38 fish, 16.58±0.169 (SE) g, and were fed twice a day for 6 weeks. The supplemented enzymes did not improve weight gain in fish fed lupin‐based diets. However, mixed enzyme significantly improved Protein Efficiency Ratio (PER). Apparent digestibility of DM, CP and GE significantly improved in fish‐fed L(E) diet. None of the supplemented enzymes affected digestive tract indices or carcass composition. Surprisingly, weight gain was significantly higher in fish‐fed L(α), L(E) and L(Mix) diets as compared with FM diet. Feed intake was significantly higher in fish‐fed L, L(α) and L(E) diets compared with the FM diet. It is concluded that storing of lupin kernel under a suitable condition may have partially hidden the positive effects of exogenous enzymes through activating the endogenous enzymes.     

The degree of carotenoid esterification influences the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

The absorption of astaxanthin from diets (30 mg kg^?1 inclusion) supplemented with either unesterified astaxanthin; isolated astaxanthin monoesters, diesters or a cell‐free carotenoid extract from Haematococcus pluvialis were studied in rainbow trout (>200 g). No significant differences (P > 0.05) were recorded in the apparent digestibility coefficients (ADC) (≈60–65%) between astaxanthin sources. However, following consumption of a single meal, peak serum astaxanthin levels at 32 h (≈1.0–1.6 μg mL^?1) were significantly higher (P < 0.05) in fish fed unesterified astaxanthin and astaxanthin monoester, compared to fish fed astaxanthin diester and the cell free extract. However, no significant differences (P > 0.05) were recorded in serum astaxanthin uptake rates between sources of astaxanthin. Results suggest that the extent of carotenoid esterification negatively influences the peak serum levels of astaxanthin in rainbow trout.     

Nutritional responses in rainbow trout (Oncorhynchus mykiss) fed diets with different physical qualities at stable or variable environmental conditions

Rainbow trout with initial body weight 1144 g were fed two diets with high (feed A) or low (feed B) water stability for 6 weeks. During the last 2 weeks either stable or fluctuating oxygen saturation, salinity and temperature was introduced. High water stability of the feed was associated with harder pellets, less dust formation and less broken pellets compared to feed with low water stability. During the first 4 weeks, feed intake was 23% higher in trout fed diet B than in those fed diet A. Fluctuating environment resulted in a severe drop in feed intake for both dietary groups. At termination of the trial, stomachs of trout fed feed B contained mashed pellets and free water and oil. Stomachs of trout fed feed A contained more intact pellets and little fluid. The apparent digestibility of protein, starch, dry matter and energy was highest in feed A. The apparent digestibility of P and Zn was highest at stable environment, and the absorption of P was highest from feed A. In conclusion, physical quality affected the nutritional value of the feeds, and differences in nutritional value at stable or fluctuating environment appeared to be related to feed intake.     

Replacement of fish meal with two fermented soybean meals in diets for rainbow trout (Oncorhynchus mykiss)

The present study investigated the effect of fish meal (FM) replacement with fermented soybean meal (FSM) on growth and feed utilization of rainbow trout. Two FSM products, FSM1 (more fermentation with more small peptide and acid than FSM2) and FSM2 were used to replace 20%, 40% and 60% of FM in control diet (250 g/kg FM), respectively (FSM1‐20, FSM1‐40, FSM1‐60, FSM2‐20, FSM2‐40 and FSM2‐60). Then the seven diets were fed to rainbow trout (18.1 g) for 8 weeks. Weight gain (WG), feed conversion ratio (FCR) and digestibility of crude protein and dry matter showed no significant difference among the groups of FSM1‐20, FSM1‐40, FSM2‐20, FSM2‐40 and the control, but WG significantly decreased and FCR increased when 60% FM was replaced by both FSMs (p < .05). The replacement of 40%, 60% FM resulted in lower villus height than the control (p < .05), and intestinal protease activity was lower in FSM2‐40, FSM2‐60 and FSM1‐60 groups than the control group (p < .05). In addition, the activity of alkaline phosphatase and superoxide dismutase increased with increasing levels of FSM (p < .05). In conclusion, dietary fish meal could be replaced by 40% with both FSMs without adverse effects on growth and feed utilization of rainbow trout based on an eight weeks feeding trial.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

Substitution of fish meal by sesame oil cake (Sesamum indicum L.) in the diet of rainbow trout (Oncorhynchus mykiss W.)

The present study evaluated the nutritional value of sesame oil cake (SOC) in rainbow trout fry (initial body weight of 1.42 g) in a growth trial performed for 45 feeding days at 15 ± 1 °C. A series of five isonitrogenous (380 g digestible protein kg^?1 dry matter (DM)) and isoenergetic (18.1 MJ digestible energy kg^?1 DM) diets were formulated in which the digestible SOC protein progressively replaced 0%, 13%, 26%, 39% and 52% of the digestible protein of a high quality fish meal (D0‐D52). Growth rate of fry significantly improved in fish fed SOC diets compared to the fishmeal control diet (D0) whatever the SOC inclusion level. This positive effect on growth was mainly related to a marked improvement of voluntary feed intake. The decrease of feed efficiency observed with increasing SOC was entirely explained by the reduction of DM and energy digestibilities with SOC incorporation. Nitrogen retention efficiency (nitrogen gain/nitrogen intake) was high (40–41%) and significantly reduced only in fish fed D52 (37%). Our results suggest that SOC can be a suitable protein source for a carnivorous fish and replace at least half of the fishmeal protein (without amino acid supplementation) without growth reduction in rainbow trout fry.     

Dietary formic acid enhances apparent digestibility of minerals in rainbow trout, Oncorhynchus mykiss (Walbaum)

The effect of dietary formic acid on the availability of phosphorus (P) from a fishmeal-based diet by rainbow trout, Oncorhynchus mykiss (Walbaum), reared in brackish water (5–6 g L⁻¹) was determined. Diets containing a low amount of P (6.0 mg P g⁻¹ dry matter) were acidified with 0, 4 and 10 mL kg⁻¹ formic acid and fed to trout (average weight, 520 g) for 4 weeks. The measured pH of the diets were 6.3, 5.8 and 5.3, respectively. The intestinal solubility of P and the digestibility of P were measured by stripping the faecal matter from the fish. The apparent digestibility coefficient (ADC) of P significantly ( P < 0.05) increased from 69.5% to 75.0% of the basal diet in fish fed diets containing 10 mL kg⁻¹ formic acid. The solubility of P in the intestine was highly variable within each treatment, and the differences were not significant. The pH of intestinal content increased with the increase in dietary formic acid concentration. The ADC of magnesium and calcium also showed a significant ( P < 0.05) increase with the acidification of diet by formic acid.     

Dietary modulation of digestive enzymes by the administration of feed additives to rainbow trout,Oncorhynchus mykiss Walbaum

The aim of this study was to assess the metabolic and digestive enzyme activities in rainbow trout, Oncorhynchus mykiss Walbaum fed dietary supplements. The biometric indices and the following enzymes activities were measured: acid protease and pepsin from the stomach homogenates, alkaline phosphatase (AP) from the small intestine and the brush border membrane, total proteolytic enzymes and trypsin from the small intestine and liver/pancreas. Dietary garlic induced a higher pepsin activity in the stomach than lipopolysaccharide (LPS) and ginger. Conversely, dietary ginger and LPS showed a significant difference (P > 0.05) in acid protease activity compared to the controls. There was not any significant difference observed between treated groups and the controls for hepato‐pancreas proteolytic enzyme activity, except for trypsin. AP activity increased significantly with dietary garlic, ginger and LPS compared to the controls.     

Identification of a novel gill-specific calpain from rainbow trout (Oncorhynchus mykiss)

Calpains are calcium-dependent neutral proteases responsible for many cellular functions. The two forms of calpain ubiquitously expressed in mammalian tissues are known as μ-calpain and m-calpain. We report here the identification of a novel calpain that is similar to but distinct from the μ- and m-calpains in rainbow trout. The cDNA of the novel gene is 2623 bp in length with a single open reading frame. The predicted protein (676 amino acids) contains the conserved calpain characteristic domains that include: domain I (pro peptide), II (cysteine catalytic site), III (electrostatic switch), and IV (calmodulin-like) with five Ca²⁺-binding EF hands. Northern blot and RT-PCR analyses demonstrated that the novel calpain gene is predominantly expressed in rainbow trout gills. Comparison of the novel protein with the ubiquitously expressed calpains and several mammalian tissue-specific calpains revealed that the novel calpain is an orthologue of the mammalian digestive tract specific calpain (calpain 9).