Effects of prebiotics on nutrient digestibility of a soybean‐meal‐based diet by red drum Sciaenops ocellatus (Linnaeus)

The present study examined the effects of four prebiotic compounds on nutrient and energy digestibility of soybean‐meal‐based diets by red drum (Sciaenops ocellatus). The experimental diets contained 40% crude protein of which approximately half was provided by soybean meal with the remainder from menhaden fish meal. The four prebiotics GroBiotic^®‐A (a mixture of partially autolysed brewers yeast, dairy ingredient components and dried fermentation products), mannanoligosaccharide (MOS), galacto‐oligosaccharide (GOS) and inulin were individually added to the basal diet at 1% by weight. A diet with all its protein provided by menhaden fish meal was also included as a control. This control diet had the highest apparent digestibility coefficient (ADC) values – 87% for protein, 87% for lipid, 78% for organic matter and 83% for energy. The basal soybean‐meal‐based diet supplemented with GroBiotic^®‐A, GOS and MOS had significantly (P<0.05) increased protein (82%, 82%, 82% respectively) and organic matter ADC values (69%, 64%, 66% respectively), compared with the basal diet (69% for protein and 49% for organic matter). However, the lipid ADC values were significantly decreased for fish fed with the diets supplemented with inulin, GOS and MOS (63%, 61%, 61% respectively) compared with the basal diet (77%) but not for those fed GroBiotic^®‐A (82%). Energy ADC values were also increased in fish fed with the GroBiotic‐A^®, GOS and MOS diets (73%, 70%, 72%), compared with the basal diet (57%); however, fish fed with the inulin diet had an energy ADC value (54%) similar to that of fish fed with the basal diet. Thus, the present study is the first to demonstrate that nutrient and energy digestibility of soybean‐meal‐based diets by red drum can be enhanced with prebiotic supplementation.     

Effects of the Prebiotics GroBiotic®‐A and Inulin on the Intestinal Microbiota of Red Drum,Sciaenops ocellatus

Two separate feeding trials examined the effects of dietary supplementation of the prebiotics GroBiotic^®‐A and inulin on growth performance and gastrointestinal tract microbiota of the red drum, Sciaenops ocellatus. In the first feeding trial, fish meal‐based diets without prebiotics or supplemented with either GroBiotic^®‐A or inulin at 1% of dry weight were fed to triplicate groups of juvenile red drum (initial weight of 2.6 g) in 110‐L aquaria operated as a brackish water (7 ppt) recirculating system for 8 wk. In the second feeding trial, soybean meal/fish meal‐based diets supplemented with either GroBiotic^®‐A or inulin at 1% of dry weight were fed to triplicate groups of red drum (initial weight of 15.8 g) in 110‐L aquaria operated as either a common recirculating water system or closed system with individual biofilters (independent aquaria) for 6 wk. Supplementation of the prebiotics in either feeding trial did not alter weight gain, feed efficiency ratio, or protein efficiency ratio of red drum fed the various diets. In the second feeding trial, the culture system significantly affected weight gain, feed efficiency ratio, and protein efficiency ratio although there were no effects of dietary treatments on fish performance or whole‐body protein, lipid, moisture, or ash. Denaturing gradient gel electrophoresis (DGGE) analysis of the gastrointestinal tract microbial community showed no effect of the dietary prebiotics as the microbial community appeared to be dominated by a single organism with very low diversity when compared with other livestock and fish species. DGGE of the microbial community in the biofilters of the independent aquariums showed a diverse microbial community that was not affected by the dietary prebiotics.     

Nutrition of red drum,Sciaenops ocellatus L.: An additional evaluation of the effects of soya‐based diets and supplemental prebiotic

We assessed the effects of a fishmeal (FM)‐ and two soya‐based diets [without or with prebiotic (GroBiotic^® ‐A; GBA) supplementation] on the production performance, non‐specific immunity, blood parameters and gut microbiota diversity of red drum, Sciaenops ocellatus. All diets were formulated to contain 450 g/kg crude protein (CP) and 120 g/kg lipid. A combination of soya bean meal and soya protein concentrate (SP) was used to replace 88% of the FM in the FM‐based diet (REF) originating the first soya‐based diet (SOY). The second soya‐based diet (SOY + GBA) was designed identically to SOY except for the supplementation of GBA (20 g/kg) substituting for wheat flour. After feeding the experimental diets to triplicate groups of 25 (~3.4 g/fish) and 90 (~4.1 g/fish) red drum juveniles for 9 (Trial I) and 16 weeks (Trial II), our results showed that (i) red drum was highly sensitive to the soya‐based diets in the first week of feeding; (ii) replacing 88% of FM with SP reduced the production performance of red drum without affecting intestinal micromorphology; (iii) whole‐blood leucocyte oxidative radical production was unaffected while lower haematocrit and higher plasma lysozyme activity were found in fish fed REF; (iv) plasma osmolality was unaffected while lower glucose and higher lactate levels were observed in fish fed SOY and SOY + GBA; (v) 16S rRNA gene‐based diversity of fish gut microbiota was affected both by SP and by GBA. This study expands the information on red drum responses to soya‐based diets and prebiotic supplementation.     

Improved digestibility of plant ingredient‐based diets for European seabass (Dicentrarchus labrax) with exogenous enzyme supplementation

This study evaluated the effect of supplementation of a plant ingredients‐based diet with two levels of commercially digestive enzyme products: Synergen? and Natugrain^® on the apparent digestibility coefficients (ADCs) of nutrients and on digestive enzyme activities in European seabass (Dicentrarchus labrax) juveniles. A control diet was formulated with 450 g/kg CP (200 g/kg from fish meal + 800 g/kg from plant protein) and 180 g/kg CL (fish oil), and four other diets were made similar to the control but supplemented with 0.2 g/kg or 0.4 g/kg of Synergen? and Natugrain^®. Digestive enzyme activities were not affected by dietary treatments. Nevertheless, chyme pH along the intestine of fish fed the Natugrain^®‐supplemented diets was reduced. The ADC of nutrients increased with exogenous digestive enzyme supplementation, with the highest ADC values being observed for the 0.4 g/kg Natugrain^® diet. The ADC of dry matter, protein, lipids, energy and phosphorus in this diet were 26%, 6%, 4%, 16% and 63% higher, respectively, than in the control diet. Overall, diet supplementation with exogenous enzyme complexes seems to have high potential for improving the utilization of plant feedstuff‐rich diets for seabass juveniles. This study revealed that Natugrain^® was more effective than Synergen? in improving the ADC of nutrients and energy in European seabass juveniles.     

The effect of exogenous enzyme supplementation on growth performance and digestive enzyme activities in Oreochromis mossambicus fed kikuyu‐based diets

The utilization of plant‐based diets in fish is limited by the presence of anti‐nutrients, which bind to dietary nutrients and reduce their bioavailability. Exogenous enzyme supplementation may alleviate the effects of anti‐nutrients and improve feed utilization. In this study, a commercial multi‐enzyme Natuzyme50^® was added to a kikuyu‐based diet at a rate of 0 (control), 0.25, 0.5, 0.75 and 1.00 g kg^?1 DM. Each diet was fed to triplicate groups of Oreochromis mossambicus for 60 days. All fish fed diets containing Natuzyme50^® had higher (P < 0.05) growth performance compared with those fed the control diet. Fish fed the diet containing 0.50 g kg^?1 had the best growth performance, highest protein digestibility and the highest levels of digestive enzyme activities. When Natuzyme50^® was supplemented above 0.50 g kg^?1 in the diet, growth performance and enzyme activities decreased. The improved growth performance observed with Natuzyme50^® supplementation was attributed to the presence of enzymes such as cellulase, xylanase and phytase in the cocktail that are not naturally produced by fish. The presence of these enzymes released bound nutrients that would have not been available to the fish. Natuzyme50^® supplementation also increased (P < 0.05) the activities of endogenous enzymes, improving the efficacy of the digestive process. The optimal dietary level of Natuzyme50^® for optimal growth performance in O. mossambicus was 0.62 g kg^?1 DM feed. Cost benefit analysis indicate that adding up to 0.5 g Natuzyme50^® kg^?1 in the diet results in higher profits as the increased growth attained offset the additional cost of Natuzyme50^®.     

Effects of dietary mannan oligosaccharide on the survival,growth, immunity and digestive enzyme activity of freshwater crayfish,Cherax destructor Clark (1936)

The aim of this experiment was to determine the effects of dietary supplementation of Bio‐Mos^® as a source of mannan oligosaccharide (MOS) on the growth, survival, immunity and digestive enzyme activity of freshwater crayfish, Cherax destructor (yabby). A basal diet and another diet with 0.4% of Bio‐Mos^® supplemented to the basal diet were formulated. Each diet was provided to eight yabbies (35.14 ± 0.48 g, initial weight) replicated 3 times in individual 250‐L plastic cylindrical tanks for a duration of 56 days. Each tank was provided with an independent recirculating water system filtered with a biological filtration. Growth parameters such as weight, specific growth rate and average weekly gain of Bio‐Mos^®‐fed yabbies were higher (P < 0.05) than yabbies fed the control diet. Total haemocyte count, granular cells and semi‐granular cells were also higher in the yabbies fed Bio‐Mos^® diet. Protease activity was higher in hepatopancreas, and amylase activity was higher in the guts of yabbies fed Bio‐Mos^® diet. The results implies that supplementation of MOS through Bio‐Mos^® can effectively improve growth and immunological condition of yabbies.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

Effects of dietary administration of Macsumsuk® on growth and stress to low salinity and low dissolved oxygen in whiteleg shrimp,Litopenaeus vannamei (Boone, 1931) juveniles

An 8‐week study was conducted to explore the results of Macsumsuk^® as a feed additive on the stress tolerance and growth of Litopenaeus vannamei in 15 culture tanks of 36 L each. Three hundred shrimp averaging 0.1 ± 0.01 g were fed with five isonitrogenous (48.38 ± 0.38% CP) diets (in triplicate groups) containing kaolinite (Macsumsuk^®) at 0%, 0.3%, 0.6%, 1.2% and 2.4%, namely Mk₀, Mk_0.3, Mk_0.6, Mk_1.2 and Mk_2.4. Specific growth rate (SGR) and weight gain (WG) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diet Mk₀ (p < .05). However, SGR and WG of shrimp fed diets Mk_0.6, Mk_1.2 and Mk_2.4 were not significantly different. Protein efficiency ratio (PER) and feed efficiency (FE) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diets Mk₀, Mk₀.₃ and Mk_0.6. Furthermore, the survival of shrimp fed diet Mk_2.4 was significantly lower than that of shrimp fed diet Mk_0.6 (p < .05). Cumulative mortality of shrimp fed diet Mk_1.2 was significantly lower than that of shrimp fed diet Mk₀ at 1–1.5 hr post‐stress to low dissolved oxygen (from 6.1 mg/L to 2.9 mg/L) and 4–5 hr post‐stress to low salinity (from 32‰ to 1‰) (p < .05). The optimum dietary Macsumsuk^® level for juvenile L. vannamei was determined as 1.97% by the polynomial regression analysis of weight gain.     

A preliminary study on the effects of dietary supplementation of brewers yeast and nucleotides, singularly or in combination, on juvenile red drum (Sciaenops ocellatus)

This study was designed to evaluate the effects of dietary supplementation with brewers yeast (Brewtech^®) and nucleotides (Optimûn), either singularly or in combination, on red drum growth, body composition, stress responses and possible resistance to Amyloodinium ocellatum infection. Juvenile red drum (～1 g initially) fed practical fish‐meal‐based diets for 6 weeks had similar weight gain regardless of whether the diet was unsupplemented (basal) or supplemented with brewers yeast (2% of diet), nucleotides (Optimûn at 0.2% of diet) or both brewers yeast (2% of diet) and nucleotides (Optimûn at 0.2% of diet). Dietary effects on hepatosomatic index, intraperitoneal fat ratio or whole‐body composition were not significant, except that fish fed the singular nucleotide supplement had a significantly higher whole‐body lipid content compared with fish fed the basal diet. No significant dietary effects on cortisol response were observed after a 15 min confinement stress test perhaps because of the extreme variation among individual fish. Subsequently, in situ challenge by co‐habitation with A. ocellatum caused 100% mortality regardless of dietary treatment within a 48 h period. It is concluded that dietary supplementation of brewers yeast and nucleotides did not promote enhanced growth or improved cortisol response and resistance to uncontrolled amyloodiniosis in juvenile red drum, at least at the tested dosages.     

Growth, reproductive performance, muscle and egg composition in grass carp, Ctenopharyngodon idella (Valenciennes), fed hydrilla or formulated diets with varying protein levels

Growth, reproductive performance, muscle and egg composition were investigated in grass carp, Ctenopharyngodon idella (Valenciennnes), fed hydrilla or formulated diets with varying protein levels. Five experimental diets, with varying levels (20%, 25%, 30%, 35% and 40%) of crude protein (CP), were used. One of the fish groups was fed hydrilla. Fish (44.1±0.3 cm; 913±9 g) were stocked (20 tank^?1) in outdoor concrete tanks (20 × 10 × 1.5 m) in duplicate, and fed to satiation, twice daily, at 09:00 and 17:00 hours for the experimental duration of 360 days. High (P<0.05) weight gain was recorded in fish fed 30% and 35% CP diets. However, values for gonadosomatic index (GSI), egg diameter, relative fecundity (eggs kg^?1 body weight), fertilizability and hatchability (%) were comparable (P>0.05) in fish at ≥25% of dietary protein intake. Hydrilla‐fed fish exhibited lower (P<0.05) values for the measured parameters. Crude protein content in muscle increased with dietary protein level. Highest (P<0.05) muscle protein was obtained in fish fed 35% CP diet. Muscle fat was comparable (P>0.05) among fish receiving formulated diets. Ash content was not significantly (P>0.05) different among fish of different dietary groups. Moisture content in fish fed formulated diets, with the exception of 20% CP diet, did not vary significantly (P>0.05). Eggs of fish fed formulated diets contained higher CP and fat contents than those of hydrilla‐fed fish. High (P<0.05) moisture content was noted in the eggs of hydrilla‐fed fish. Ctenopharyngodon idella fed formulated diet, with a minimum of 25% CP, showed better reproductive performance than those fed hydrilla.     

Evaluation of dietary natural mineral materials as an antibiotic replacer on growth performance,non‐specific immune responses and disease resistance in rainbow trout,Oncorhynchus mykiss

We evaluated the effects of some dietary natural mineral materials as an antibiotic replacer based on growth performance, non‐specific immune responses and disease resistance in juvenile and subadult rainbow trout, Oncorhynchus mykiss. First experiment, juvenile rainbow trout averaging 2.7 ± 0.02 g (mean ± SD) were fed one of the six experimental diets; a basal commercial diet as a control (CON), CON with oxytetracycline (OTC), with yellow loess (YL), with Macsumsuk^® (MS), with Song‐Gang^® stone (SG) and with barley stone (BS) at 0.4% of each diet. At the end of 8‐week feeding trial, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) of fish fed YL diet were significantly higher than those of fish fed CON diet. Non‐specific immune responses such as superoxide dismutase (SOD), myeloperoxidase (MPO), lysozyme (LYS) activity and oxidative radical production of fish fed YL diet were higher than those of fish fed CON diet. At the end of 15 days of challenge test with Aeromonas salmonicida, average cumulative survival rate of fish fed YL diet was significantly higher than that of fish fed BS and CON diets. However, there were no significant differences among fish fed YL, SG and OTC diets. Second experiment, subadult rainbow trout averaging 261.5 ± 3.5 g (mean ± SD) were fed one of the four experimental diets for 22 weeks: CON, and CON with OTC, YL or SG at 0.4% of each diet. At the end of feeding, growth performance of fish fed SG and YL diets was significantly higher than that of fish fed CON diet. Non‐specific immune responses in terms of SOD, MPO, LYS and NBT of fish fed SG and YL diets were significantly higher than those of fish fed CON diet. However, there were no significant differences among the fish fed YL, SG and OTC diets. The results indicate that dietary yellow loess or Song‐gang^® stone at 0.4% of diet could replace oxytetracycline in juvenile and subadult rainbow trout.     

Combined effect of dietary protein and phytase levels on growth performance,feed utilization,and nutrients digestibility of African catfish,Clarias gariepinus (B.), reared in earthen ponds

The present study was based on a 2 × 4 factorial design with two levels of dietary protein (33% or 37% CP) and four phytase levels (0.0, 750, 1,000, and 1,250 FTU/kg diet). African catfish, Clarias gariepinus (B.) (10.7 ± 0.4 g), were distributed into earthen ponds (100 m² surface area per each) at a density of 4.5 fish per m². Fish were fed on the experimental diets up to satiation twice a day for 3 months. Another experiment with the same treatments was conducted in 45-L tanks for 2 months to evaluate nutrient retention and digestibility. The growth and production of African catfish fed phytase-enriched diets were higher than those fed the control diet. The highest fish performance and production was observed with fish fed 37% CP enriched with a 1,200 FTU/kg diet of phytase. Feed intake was significantly affected by supplemental phytase alone, where it increased significantly as dietary phytase increased at both protein levels, resulting in similar FCR values (1.31–1.46). Additionally, the maximum values of protein efficiency ratio, protein retention, and phosphorus retention were obtained at 33% CP with phytase levels of 750–1,000 FTU/kg diet. It is also noticed that organic matter, protein, and phosphorus were more digestible in fish fed a 37% CP diet with high phytase levels. These results suggest that the optimum performance and production of African catfish were observed at a 37.0% CP diet enriched with a phytase level of 1,200 FTU/kg diet.     

Optimum Dietary Protein Level and Protein‐to‐energy Ratio for Growth of Juvenile Parrot Fish,Oplegnathus fasciatus

An 8‐wk feeding trial was conducted to estimate the optimum dietary protein level and protein‐to‐energy (P/E) ratio in juvenile parrot fish, Oplegnathus fasciatus. Eight experimental diets were formulated with two energy levels and four protein levels for each energy level. Diets containing crude protein (CP) at 35, 40, 45, and 50% had either 12.5 or 14.6 kJ/g of energy. Fish averaging 7.1 ± 0.06 g (mean ± SD) were fed one of the experimental diets for 8 wk. At the end of the feeding trial, weight gain (WG) of fish fed 45 and 50% CP in the 12.5 kJ/g diet was significantly higher than fish fed the 35% CP diet (P < 0.05). WG of the fish fed 45 and 50% CP in the 14.6 kJ/g diet was significantly higher than fish fed the 35 and 40% CP diets (P < 0.05). Fish fed the 14.6 kJ/g diet had a higher WG compared with fish fed the 12.5 kJ/g diet at all CP levels. Feed efficiency (FE) and specific growth rate (SGR) showed a similar trend to the WG. WG, FE, and SGR improved with increasing dietary protein levels up to 45% and remained constant at 50% CP for both energy levels. However, protein efficiency ratio was negatively related to dietary protein levels. The results suggested that the optimum level of protein and the optimum P/E ratio for juvenile parrot fish should be 45% and 31.1 mg protein/kJ, respectively, in a diet containing 14.6 kJ/g energy.     

Immune Response and Resistance to Stress and Edwardsiella ictaluri Challenge in Channel Catfish, Ictalurus punctatus, Fed Diets Containing Commercial Whole-Cell Yeast or Yeast Subcomponents

Dietary supplementation of yeast or yeast subcomponents (YYS) as commercial preparations of β‐glucan (MacroGard^®; Biotec‐Mackzymal, Tromsø, Norway; and Betagard A^®; Aqua‐In‐Tech, Inc., Seattle, WA, USA), mannan oligosaccharide (Bio‐Mos? Aqua Grade; Alltech, Nicholasville, KY, USA), or whole‐cell Saccharomyces cerevisiae (Levucell SB20^®; Lallemand Animal Nutrition, Milwaukee, WI, USA) at the manufacturer’s recommended levels was evaluated on the physiological performance of juvenile channel catfish, Ictalurus punctatus. Fish were fed YYS diets for 4 wk, followed by 2 wk of control diet. Fish were sampled at the end of each feeding period (4 and 6 wk) to measure hematological and immune parameters and growth and to determine the effects of dietary β‐glucan on resistance to Edwardsiella ictaluri infection and to low‐water stress (6 wk). Supplementation of YYS in diets did not affect growth performance, hematology, or immune function. Survival from E. ictaluri infection was from 5 to 17.5% higher in fish fed YYS diets than in the control group, but the increases were not significant. Some improvement in stress resistance was observed in YYS‐fed catfish after exposure to low‐water stress. Stress reduction in fish fed diets supplemented with yeast subcomponents has been reported previously, but thus far, no explanation has been proposed for this effect. The present study and the previously published research suggest that dietary YYS supplementation does not appear to improve resistance of channel catfish to E. ictaluri.     

Salt in the Food and Water as a Supportive Therapy for Ichthyophthirius multifiliis Infestation on Silver Catfish, Rhamdia quelen, Fingerlings

This study verified the efficacy of common salt (NaCl) in the food and water as a supportive therapy for Ichthyophthirius multifiliis and growth of silver catfish, Rhamdia quelen, fingerlings. After appearance of the first white spots following experimental exposure, indicating infestation by this parasite, fish were transferred to eighteen 40‐L polypropylene tanks and kept for 30 d. Experimental diets were prepared with five different levels of salt supplementation: 0.0, 1.2, 2.5, 5.0, and 6.0% for 30 d. In addition, another group of fish were fed a diet without salt supplementation but exposed to 4 g of salt/L of water. Fingerlings were collected at different times for analyses of whole‐body Na⁺, K⁺, and Cl^?. Dietary salt supplementation provoked a transitory increase in whole‐body Na⁺, K⁺, and Cl^? in infested catfish fingerlings, but the best stabilization of these ion levels was observed in specimens exposed to salt in the water. Silver catfish fed a diet containing 1.2% salt showed higher weight and biomass after 30 d compared to those fed with 6% salt. The use of 4 g of salt/L of water improved survival of infested silver catfish, but dietary salt supplementation was ineffective as a therapy for ichthyophthiriasis.     

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.     

Supplementation with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa) in low fish meal diets for the white shrimp,Litopenaeus vannamei

This work evaluated the performance of Litopenaeus vannamei to low fish meal diets supplemented with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa). A basal diet with 150.0 g kg^?1 of anchovy fish meal was designed. Two positive control diets were formulated to reduce fish meal at 50% and 100% with 1.0 and 2.0 g kg^?1 of MERA? Met_Ca (calcium salt with 84% HMTBa activity), respectively. Two nearly equivalent diets acted as negative controls, without HMTBa supplementation. A total of 50 clear‐water tanks of 500 L were stocked with 2.22 ± 0.19 g shrimp under 70 animals m^?2. Shrimp survival (92.3 ± 5.1% and 81.4 ± 8.0%), yield (808 ± 12 and 946 ± 17 g m^?2) and FCR (2.17 ± 0.19 and 3.12 ± 0.37) showed no differences among diets after 72 or 96 days, respectively. A significantly higher shrimp body weight and weekly growth were observed for those fed with the basal diet or diets supplemented with HMTBa compared with non‐supplemented ones. This study has shown that L. vannamei growth, body weight, survival, yield and FCR were supported by HMTBa supplementation when 150.0 g kg^?1 of fish meal was replaced by soybean meal and other ingredients, at 50% and 100%.     

Effects of dietary supplementation of a commercial prebiotic Previda® on survival,growth, immune responses and gut microbiota of Pacific white shrimp,Litopenaeus vannamei

A 35‐day feeding trial was conducted to evaluate growth, bacterial populations of the gastrointestinal tract (GIT) and immune responses of Litopenaeus vannamei fed diets containing the commercial prebiotic Previda^®. Diets were formulated to contain Previda^® at 0, 0.2, 0.5, 1.0 or 1.6 g kg^?1 by weight. At the end of the study, differences in weight gain and survival among treatments were not significant (P > 0.05), but denaturing gradient gel electrophoresis analysis revealed that the microbial communities in the GIT changed significantly with the inclusion of dietary Previda^® at different levels. Previda^® was therefore able to selectively modify the microbial communities in the shrimp's GIT. Although individual bacterial species were not identified, enteric populations in shrimp fed the prebiotic at similar levels of inclusion were genetically similar. In addition, shrimp fed Previda^® at 1.6 g kg^?1 responded significantly (P < 0.05) better immunologically with respect to hemocyte phagocytic capacity, haemolymph protein, hyaline cell counts and haemolymph glucose compared with shrimp fed the basal diet. Although shrimp were not exposed to virulent pathogens in this study, the observed upregulation of some of imm‐une responses upon prebiotic supplementation indicates that an improved outcome of such challenges may be anticipated in Previda^®‐fed shrimp under commercial conditions.     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

Effect of dietary supplementation of inulin and W. cibaria on haemato‐immunological parameters of hybrid surubim (Pseudoplatystoma sp)

The dietary supplementation of prebiotics, probiotics and symbiotic in hybrid surubins (a Pseudoplatystoma corruscans and P. fasciatum cross) was evaluated for the effects on their autochthonous intestinal microbiota and on haematological and immunological parameters. A total of 160 fish were divided into four treatment groups with four replicates each. The treatment groups were fed with the following diets for 15 days: control diet without supplementation; 0.5% inulin (prebiotic) supplementation; Weissella cibaria (CPQBA 001‐10 DRM 02) (7.87 ± 0.2 log CFU g^?1) supplementation; or 0.5% inulin and W. cibaria supplementation (symbiotic group). The midgut intestines of the fish with the symbiotic diet supplementation had higher concentrations of lactic acid bacteria (7.07 ± 1.11 log CFU g^?1) and low levels of Vibrio spp (1.90 ± 0.60 log CFU g^?1) and Pseudomonas spp (2.23 ± 1.48 log CFU g^?1). In addition, increased erythrocytes and reduced circulating neutrophils were observed in this group. No differences in blood glucose, serum protein or lysozyme levels were detected between treatment groups. However, a higher concentration of total immunoglobulin was observed in fish fed with the probiotic and symbiotic diets. The addition of 0.5% inulin (prebiotic) thus W. cibaria (probiotic) to the diet of Pseudoplatystoma hybrid surubins reduce the number of pathogenic bacteria and stimulate the beneficial intestinal microbiota and may possibly alter their immune defence system.