The effect of insect meal as a feed ingredient on survival,growth, and metabolic and antioxidant response of juvenile prawn Palaemon adspersus (Rathke, 1837)

For successful prawn aquaculture, feeds should be based on readily consumed ingredients that promote survival and optimal growth performance. This pilot study investigates the rearing of juvenile Baltic prawn Palaemon adspersus. Two feeding trials were carried out for 60 days; both incorporated insect meals, the first one in fishmeal‐based diets, whereas the second one in plant meal ones. Insect meals derived from larvae of Tenebrio molitor (TM), Hermetia illucens (HI) and Musca domestica (MD) were tested as feed ingredients. This study indicated that the inclusion of HI in fishmeal diets resulted in significantly higher growth performance and survival of the prawns, whereas the MD diet led to similarly high growth performance reducing significantly their survival. Growth performance was not affected by the insect inclusion in the plant‐based diets, but survival was higher in the TM and HI inclusion diets. The inclusion of TM and HI resulted in higher protein and energy content of the prawns’ muscle when incorporated in fishmeal and plant meal diets respectively. No significant differences were observed in the activities of hepatopancreas’ amino acid‐catabolizing enzymes. Concluding, the combinations fishmeal–HI and plant meal–TM can be used for the successful rearing of Baltic prawn.     

Incorporation of dietary nitrogen from fish meal and pea meal (Pisum sativum) in muscle tissue of Pacific white shrimp (Litopenaeus vannamei) fed low protein compound diets

Stable isotope analyses were applied to explore the relative dietary nitrogen contributions from fish meal and pea meal (Pisum sativum) to muscle tissue of Pacific white shrimp postlarvae (141 ± 31 mg) fed low protein diets having different proportions of both ingredients as the sole dietary protein sources. A negative control diet was formulated to contain 100% pea meal and six more isoproteic diets to have decreasing levels of pea meal‐derived nitrogen: 95%, 85%, 70%, 55%, 40% and 0% of the initial level. Growth rates were negatively correlated to dietary pea protein inclusion due to progressive essential amino acid deficiencies (sulphur amino acids, threonine, lysine, histidine). The nitrogen turnover rate significantly increased in muscle tissue of shrimps fed diets having high levels of pea meal; however, contrary to observations from a previous study using soy protein, the relative contributions of dietary nitrogen from pea meal to shrimp muscle tissue were equal or higher than expected contributions established by the dietary formulations. Results highlight the effectiveness of stable isotope analysis in assessing the nutritional contributions of alternative ingredients for aquaculture feeds and the potential suitability of pea as a source of protein (provided the diets are nutritionally balanced).     

Apparent digestibility of animal,plant and microbial ingredients for Pacific white shrimp Litopenaeus vannamei

A digestibility trial was conducted to determine apparent digestibility coefficients of dry matter, protein, energy and amino acids of animal, plant and microbial ingredients for Pacific white shrimp, Litopenaeus vannamei. The tested ingredients included traditional soybean meal (TSBM), PepsoyGen soybean meal (PSBM), NutriVance soybean meal (NSBM), fish meal (FM), poultry meal (PM), squid hydrolysis (SQH), scallop hydrolysis (SCH), flash dried yeast (FDY), two batches of Ulva meal (UMF and UMS) and bacteria biomass (BB). A basal diet was formulated and produced along with the experimental diets which included 300 g/kg of each ingredient and 700 g/kg of the basal diet. Juvenile shrimp (initial mean weight: 12 g, six shrimp/tank, n = 3) were stocked in a recirculation system. Apparent dry matter, protein and energy digestibility coefficients ranged from ?40.11% to 78.51%, 15.17% to 97.03% and 13.33% to 82.56% among different protein sources, respectively. In general, protein and energy digestibilities in soy sources (77.6% to 97.03% and 62.77% to 82.56%, respectively) are higher than the tested animal protein (51.39% to 71.41% and 45.29% to 69.77%, respectively) and single‐cell protein sources (15.17% to 53.47% and 13.33% to 40.39%, respectively). Among the three soybean sources, TSBM showed highest protein and energy digestibility. Apparent individual amino acid digestibility coefficients were also variable among different types of ingredients, and there was a reasonable correspondence to protein digestibility. The most digestible feed ingredients for Pacific white shrimp in this study were conventional soybean meal (SBM) and NutriVance soybean meal (NSBM), which indicated that these ingredients are good protein and amino acid sources for Pacific white shrimp. Resultant digestibility data may provide useful information to commercial shrimp feed industry.     

Apparent digestibility coefficients of selected feed ingredients for juvenile snakehead,Ophiocephalus argus

Apparent digestibility coefficients (ADCs) of dry matter, crude protein, crude lipid, gross energy, phosphorus and amino acids in Peruvian fish meal, poultry by‐product meal, meat and bone meal, spray‐dried blood meal, hydrolysed feather meal, corn gluten meal, soybean meal, peanut meal, cottonseed meal and rapeseed meal were determined for juvenile snakehead (Ophiocephalus argus) with initial mean body weight of 78.1 g. A reference diet and test diets that consisted of a 70 : 30 mixture of the reference diet to test ingredient were used with 5 g kg^?1 Cr₂O₃ as an external indicator. Fish meal, poultry by‐product meal and corn gluten meal had higher ADCs of dry matter, crude protein, and gross energy among ingredients tested. Dry matter ADCs ranged 61.9–81.5% for animal ingredients and corn gluten meal and ranged 52.2–68.0% for soybean meal, peanut meal, cottonseed meal and rapeseed meal. Energy ADCs of ingredients followed similar trends to differences in dry matter digestibility. Protein ADCs of animal and plant ingredients ranged 73.6–92.8% and 75.3–85.6%, respectively. Amino acid ADCs generally reflected protein digestibility. Lipid ADCs were relatively high for the ingredients tested. Phosphorus ADCs of animal and plant ingredients ranged 39.5–65.2% and 38.7–57.1%, respectively.     

Digestibility of nutrients and energy in ingredients for bullfrogs during different phases of development

The ability of frogs to digest dietary nutrients changes with growth, with the animals becoming more or less demanding. The objective of this study was to determine the apparent digestibility coefficients of protein, energy and ether extract of 14 ingredients used in bullfrog diets (spray‐dried blood meal, hydrolysed feather meal, poultry by‐product meal, red blood cell concentrate, tilapia by‐product meal, sardine fishmeal, salmon fishmeal, meat and bone meal, corn gluten meal, soybean meal, corn, wheat bran, soy protein concentrate and soybean oil). A total of 2,325 bullfrogs (Lithobates catesbeianus) in different phases of development were used: early phase (mean weight of 30–50 g), growth phase (80–110 g) and finishing phase (150–200 g). Faeces were collected using the method of dissection. Marked differences in digestibility of the ingredients were observed between the different phases of frog development. Among the ingredients studied, salmon fishmeal and corn gluten meal showed good utilization of the protein (78.9% and 86.7%, respectively) and energy fraction (89.4% and 83.3%, respectively). The salmon fishmeal, poultry by‐product meal, sardine fishmeal, soy protein concentrate, wheat bran and soybean oil exhibited good ether extract utilization (81.2%–92.8%), recommending their use in bullfrog diets.     

Assessment of the effect of plant inhibitors on digestive proteases of nile tilapia using in vitro assays

This study was conducted to evaluate the effect that feeding on differently treated soybean meals may have on digestive protease activity of the Nile tilapia (Oreochromis niloticus). Untreated full fat soybean (USB), heated fullfat soybean (HSB), soaked fullfat soybean (SSB) and commercial defatted soybean meal (DSB) totally replaced fish meal (FM) in test diets (30% crude protein) fed to fish juveniles (mixed sexes) for 30 days. After this period, total activity of digestive proteases was determined in each group and the ability of such proteases to hydrolyse the different protein sources and formulated feeds was tested in vitro. Significant differences were observed between treatments, with lower values obtained for fish fed on USB, suggesting a modification in the composition of secreted proteases. In addition, an inhibitory effect produced by wheat bran included in the test diets on protease activity of fish was also shown. The usefulness of using in vitro assays for a preliminary assessment of the nutritive value of aquafeed ingredients is discussed.     

Growth,tissue metabolites and digestive functionality in Sparus aurata juveniles fed different levels of macroalgae,Gracilaria cornea and Ulva rigida

The aim of this study was to evaluate the inclusion of two macroalgae as dietary ingredients on growth, body composition, intermediate metabolism, intestinal enzymes and gut histology of Sparus aurata juveniles. Six isocaloric and isonitrogenous diets incorporating 5%, 15% or 25% of Gracilaria cornea or Ulva rigida meal were tested against a control diet. Duplicate groups of fifteen fish (14 g) were fed the experimental diets for 70 days. Growth performance showed an inverse linear relationship with the inclusion level of Gracilaria. Feed conversion ratio increased significantly in fish fed 25% Gracilaria. Ulva presented a quadratic relationship with the lowest SGR at 15% of inclusion. Fish fed 25% Ulva exhibited significantly lower lipid content than those fed control diet. Overall, metabolites did not show differences with specimens fed control diet, except for plasmatic glucose and muscle lactate. Intestinal proteolytic activity was reduced with the macroalgae inclusion level, but all fish showed the same enzyme pattern. The histological study revealed no negative effect on liver and intestine structure by the macroalgae inclusion. The present work demonstrated that these ingredients can be used in practical diets for sea bream, but a more comprehensive research is needed to determine the optimum dietary level in a longer feeding trial.     

Potential bioavailability of protein and lipids in feed ingredients for the three‐spot cichlid Amphilophus trimaculatus: An in vitro assessment

For the commercial culture of the three‐spot cichlid (Amphilophus trimaculatus), it is important to have available a set of protein and lipid ingredients suitable for feed manufacturing, which allow to optimize the culture at affordable costs. The in vitro digestibility was evaluated with pH‐stat technique, using stomach and intestine multi‐enzymatic extracts from A. trimaculatus juveniles. The digestion of protein ingredients was taken place both in acid and in alkaline conditions, quantifying the total of free amino acids at the end of the hydrolysis, whereas the digestion of lipid ingredients was taken place only in alkaline conditions. The digestive enzymes of this species showed a better affinity to raw materials of animal origin such as beef meal, fish meal and premium chicken meal, which presented better digestibility. The best lipid ingredient that could be used is the fish oil. However, it is possible to use ingredients from the plant sources, such as soy or cereals, and vegetable oils such as olive, corn or soy oil. It is necessary to complement this study in vitro with the digestibility analysis in vivo, in order to determine the inclusion percentages in feeds and costs and therefore to develop an efficient feed formulation for the aquaculture of A. trimaculatus.     

In vivo and in vitro protein digestibility and growth performance of animal and no‐conventional plant ingredients for river prawn Macrobrachium americanum (Caridea: Palaemonidae)

The objectives of this study were to evaluate the effects of non‐traditional plant and animal protein for river prawn M. americanum on in vivo versus in vitro digestibility in addition to growth performance. In this study, ten ingredients were used: coconut meal, jackfruit meal, soybean meal, poultry by‐product meal, squid meal and fish meal. In vivo studies were conducted using the zeolite marker method, whereas in vitro digestibility was determined by the pH‐stat method using river prawn hepatopancreas enzymes. According to the findings, river prawns digest animals more efficiently than plant ingredients, with squid and fish meal showing a tendency of higher values. Fish meal (53.66%), squid meal (48.52%) and jackfruit meal (42.21%) were found to have a digestibility greater than 40% in in vivo and in vitro methods, thus suggesting that they are the best ingredients used in this study. This also validates their inclusion in practical diets. The correlation of digestibility of in vivo and in vitro was low (R² = .6749). The highest daily weight gain was recorded in the diets containing jackfruit, coconut and squid. Given that the potential of no‐conventional plant ingredients available regionally was significantly higher, this underscores the need to conduct further research to validate their inclusion in practical diets.     

In vitro protein digestibility of different grow‐out stages of spotted rose snapper (Lutjanus guttatus,Steindachner, 1869)

The aim of this study was to compare in vitro protein digestibility between two groups of fish, at early (21 g) and late stages (400 g) of spotted rose snapper Lutjanus guttatus, to evaluate the degree of hydrolysis (DH) and total amino acid release (TAAR) using crude extracts from stomach, pyloric caeca and intestine of 13 protein ingredients including marine, animal and plant meals. Degree of hydrolysis and TAAR were measured by a pH‐Stat method, and the PAGE‐Zymogram was also used as complementary technique. Differences in DH were found between both grow‐out stages mainly in the alkaline hydrolysis phase. Fish and squid meals (marine sources) had the highest DH and TAAR, followed by porcine meat and poultry meal by‐products from recycling sources, and soybean and canola meals (plant sources), which represent better protein sources for use in practical diets. Stomach zymograms showed two pepsin isoforms in both grow‐out stages. Pyloric caeca and intestine zymograms showed five bands with proteolytic activity in the early grow‐out stage, whereas four additional bands were found in late grow‐out stage. Alkaline proteases were identified as serine and metalloproteases. Thus, L. guttatus presents an ontogenetically differentiated digestive enzyme pattern that modifies the DH and TAAR of different protein sources.     

Effect of fungal fermentation on apparent digestibility coefficient for dry matter,crude protein and amino acids of various plant protein sources in Penaeus vannamei

Four commercial solvent‐extracted oilseed meals/cakes, viz. soybean meal (SBM), groundnut oil cake (GNC), rapeseed meal (RSM) and sunflower oil cake (SFC), were fermented with fungus Aspergillus niger, and its effect on apparent digestibility coefficient (ADC) was studied in Penaeus vannamei. Reference diet and eight experimental diets containing 700 g/kg reference diet and 300 g/kg test ingredient with 5 g/kg chromic oxide were formulated. Each diet was randomly allotted to three tanks containing ten shrimps. Shrimps were adapted to the experimental diets for a week, and faeces were collected using Falcon tube from second week onwards. The ADC of all the ingredients significantly (p < .05) increased with fermentation and the increase being higher in SBM (78.46%–91.71%) for dry matter and in SFC (71.51%–87.02%) for protein. Analysis of variance showed that the ADC of both dry matter and protein significantly (p < .05) differed in treatments (p = <.001) and ingredients (p = <.001). The average ADC of ingredients was ranked as SBM > GNC > RSM > SFC. The most digestible essential amino acid (EAA) in fermented ingredients was methionine in SBM, arginine in GNC, valine in RSM and histidine in SFC. A better improvement in amino acid digestibility was observed in fermented SFC. Results indicated that P. vannamei efficiently digests fermented ingredients compared to unfermented ones.     

Advances in the study of activity additivity of supplemented proteases to improve digestion of feed protein by Penaeus vannamei

In vitro assays used porcine or bovine trypsin as models of exogenous enzymes to determine functioning in the presence of enzymatic extracts from the digestive gland of whiteleg shrimp Penaeus vannamei. Using electrophoresis and zymograms, when enzymes from the shrimp were mixed in the absence of protein substrate, they hydrolysed the trypsin from bovine or porcine origin. Porcine or bovine trypsin, when mixed with shrimp enzymes in pH‐stat assays in the presence of shrimp commercial feed, fish meal, or casein, there was added activity to hydrolyse the protein substrate. Hydrolysis of protein substrate was twofold to threefold stronger if exogenous enzymes were added. Results suggest that porcine or bovine trypsin could be used as feed supplements for whiteleg shrimp P. vannamei to enhance hydrolysis of proteins in feeds, because the commercial enzymes contributed to the hydrolysis of the protein in the three substrates in the presence of shrimp enzymes.     

Growth and economic analysis of freshwater prawn,Macrobrachium rosenbergii (de Man), produced with feeds substituting sunflower cake for fish meal,soya bean meal and mustard oil cake

A feeding trial with Macrobrachium rosenbergii was carried out to test the effectiveness of sunflower cake (SFC) as a replacement for conventional protein sources. Four isonitrogenous (~30% crude protein) and isocaloric (388–402 kcal) diets were formulated. Diet‐1 was a control made with conventional ingredients including fish meal, soya bean meal and mustard oil cake. Diet‐2, Diet‐3 and Diet‐4 were made with 30%, 40% and 50% SFC, respectively, replacing the usual protein sources used in Diet‐1. After 100 days, diets with SFC inclusion showed favourable growth and economic performance trends compared to control diet‐1, but without significant differences (p > .05). Better production (572.89 ± 62.99 kg/ha) was observed with Diet‐4 followed by Diet‐2, Diet‐3 and Diet‐1. Diet‐4 revealed the best feed conversion ratio (2.56 ± 0.24), specific growth rate (1.18 ± 0.05) and protein efficiency ratio (1.14 ± 0.10) among these diets. Whole carcass composition analysis revealed highest crude protein (18.19%) and lipid (4.43%) content in prawns fed Diet‐3 followed by Diet‐4. Highest total income, gross margin and benefit cost ratio (BCR) per hectare from prawns were US$5,759, US$2,623 and 1.84, respectively, found in Diet‐4. This study demonstrates that 30%–50% inclusion of SFC has favourable impacts on productivity and profitability compared with controls, with 50% SFC inclusion as the most cost‐effective diet for prawns without apparent compromise in growth, flesh quality or economics. The performances of the diets suggest that SFC may be a suitable substitute for fishmeal, soya bean meal and mustard oil cake in prawn feed. The effectiveness of higher SFC concentrations should also be investigated.     

Evaluation of Three In Vitro Enzyme Assays for Estimating Protein Digestibility in the Pacific White Shrimp Penaeus vannamei1

Abstract— In vitro enzyme assays are rapid, inexpensive techniques for estimating protein digestibility of feed ingredients. Three assays—the Lazo single-enzyme assay with porcine trypsin; the Hsu multi-enzyme assay with porcine trypsin, α-chymotrypsin, and peptidase; and the Satterlee multi-enzyme assay with porcine trypsin, α-chymotrypsin, peptidase, and bacterial protease—were used to estimate relative protein digestibility (RPD) of selected feed ingredients used in diets for the Pacific white shrimp Penaeus vannamei. Ingredients tested were casein, gelatin, rice bran, shrimp meal, soybean meal, wheat gluten, and six varieties of fish meal. A highly significant, inverse, linear relationship existed between final pH in each of the enzyme assays and in vivo apparent protein digestibility of the ingredient in P. vannamei feeding trials. Ranking of RPD coefficients obtained in the Lazo single-enzyme assay followed closely the ranking of published in vivo apparent protein digestibility coefficients for the same ingredients in P. vannamei digestibility trials. Results indicated that a single-enzyme assay with porcine trypsin identified differences in protein digestibility of feedstuffs in vitro as well as the two multi-enzyme assays tested. The porcine-trypsin, single-enzyme assay appears to be a potentially useful tool for evaluating protein quality and relative protein digestibility of feedstuffs for P. vannamei.     

Evaluation and characterization of a novel probiotic Lactobacillus pentosus PL11 isolated from Japanese eel (Anguilla japonica) for its use in aquaculture

In the present study, a potential Lactobacilli probiotics were isolated from Japanese eels (Anguilla japonica) and characterized and evaluated for their possible use in eel farming. Sixteen Lactobacilli were isolated from intestines of Japanese eels, using selective media. The lactobacilli strains (represented as PL1 to PL16) were screened by their ability to produce digestive enzyme. Among these, three strains (PL11, PL13 and PL16) producing four digestive enzymes (amylase, cellulase, protease and phytase) simultaneously were characterized further using API ZYM kit. From these, PL11 (Lactobacillu (L.) pentosus) was identified as potential probiotics candidate producing 15 enzymes among 20 tested. Further examination of biological activities of PL11 revealed tolerance against pH, artificial bile juice and antibacterial activity against several fish pathogenic bacteria. The in vitro competitive exclusion assay also revealed 88.4% reduction in adhesion of fish pathogen (Edwardsiella tarda) by PL11 to host intestinal mucus. In vitro incubation of Japanese eel foregut with Baclight‐labelled PL11 showed colonization of the enterocyte surface by confocal and scanning electron microscopy. In summary, PL11 isolated from eels could serve as a potential probiotics with acid and bile tolerance, production of digestive enzymes, antibacterial activity and inhibition of fish pathogen adhesion to intestinal mucus.     

Bioavailability of arginine from Indian mustard protein concentrate and meal compared with that of a soy protein concentrate in rainbow trout (Oncorhynchys mykiss)

Relative bioavailability (RBV) of arginine (Arg) from Indian mustard protein concentrate (IMC, 62% crude protein) and Indian mustard meal (IMM, 42% crude protein), and a commercially available soy protein concentrate (SPC, 57% crude protein) was compared with that of crystalline L‐arginine (L‐Arg) in rainbow trout. A basal diet highly deficient in Arg (1.23%) was formulated. Eight other isoproteic and isoenergetic diets were formulated to contain ~1.35% and ~1.5% Arg by adding increasing amount of IMC, IMM, SPC and L‐Arg. The experimental diets were fed for 16 weeks using a standard protocol. Growth rate, weight gain (g fish^?1) and protein (PD, g fish^?1) and lipid (LD, g fish^?1) deposition were increased linearly with increasing level of Arg from all ingredients. Arg availability from protein‐bound sources were equal or higher than those from L‐Arg. RBV of Arg from IMC, IMM and SPC were ranged from 100% and 123% than that from L‐Arg (assumed as 100% bio‐available). Among the ingredients, only the RBV of Arg from IMC was significantly higher than those from SPC (P < 0.05). The findings suggest that the RBV of Arg from IMC and IMM are very good and comparable to that of the commercial SPC used in this study.     

Amino acid digestibility of animal protein ingredients for bullfrogs in different phases of post‐metamorphic development

The development of diets that economically and ecologically meet the requirements of bullfrogs (Lithobates catesbeianus) is important for frog farming. Therefore, the objective of this study was to determine the apparent digestibility coefficients for essential amino acids of eight animal feed ingredients (poultry byproduct meal, meat and bone meal, salmon byproduct meal, tilapia byproduct meal, spray‐dried blood meal, powered red blood cells, feather meal and sardine meal) used in the diets of bullfrogs. Additionally, the limiting digestible amino acids were determined for each ingredient. Bullfrogs in different phases of post‐metamorphic development were used: 810 animals in the early phase (mean weight: 40 ± 10 g), 405 animals in the growing phase (95 ± 15 g) and 243 animals in the finishing phase (175 ± 25 g). Poultry byproduct meal, salmon byproduct meal and blood meal exhibited the largest number of high apparent digestibility coefficients for essential amino acids (>70%) in the early, growing and finishing phases, respectively. The animal protein ingredients were well digested by the bullfrogs and could be used in practical diets for this species, respecting the limits of inclusion and digestibility values for each developmental phase.     

Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings

Nine isonitrogenous (35% crude protein approximately) and isocaloric (18.37 kJ g^?1) experimental diets (RLL20–BCFL40) were formulated with either raw or treated (inoculated with fish intestinal bacteria) Leucaena leucocephala leaf meal at 20%, 30% and 40% levels replacing other ingredients partially from a fish meal based reference diet (RD). Two specific strains of fish intestinal bacteria, Bacillus subtilis (isolated from Cyprinus carpio) and B. circulans (isolated from Oreochromis mossambicus) having extracellular cellulolytic and amylolytic activities, were used to inoculate Leucaena leaf meal for 15 days at 37°C. The crude fibre, cellulose and hemicellulose contents and the antinutritional factors, tannin, phytic acid and mimosine in the leaf meal decreased due to inoculation. However, free amino acids and fatty acids increased in the treated leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a RD. Both the inclusion level and type of Leucaena leaf meal in diets significantly affected the growth performance of rohu. Fish fed diets containing inoculated Leucaena leaf meal performed better in comparison with those with the RD. On the basis of growth response, feed conversion ratio, protein efficiency ratio and apparent net protein utilization, diet formulated with 30%Leucaena leaf meal inoculated with B. circulans resulted in the best performance of rohu fingerlings followed by diet with 40%B. subtilis inoculated Leucaena leaf meal. The apparent protein digestibility (APD) was better in fish fed diets containing B. circulans inoculated leaf meal. An increasing level of raw Leucaena leaf meal was associated with a decrease in the carcass protein content of rohu fingerlings. The activity of α‐amylase increased with the increasing level of treated leaf meal in diets. Cellulase activity increased with increasing level of inclusion of raw leaf meal, and was comparatively lower in fish fed diets with treated leaf meal. Activities of protease and lipase were higher in fish fed the RD. The results showed that it is possible to incorporate Leucaena leaf meal inoculated with enzyme‐producing fish intestinal bacteria in carp diets up to 40% level of inclusion.     

Evaluation of three non‐genetically modified soybean cultivars as ingredients and a yeast‐based additive as a supplement in practical diets for Pacific white shrimp Litopenaeus vannamei

Two growth trials and a physiology assessment were conducted to evaluate three non‐genetically modified (GM) soybean cultivars as ingredients in practical diets for Pacific white shrimp, Litopenaeus vannamei. In addition, a commercially available fermented yeast product was evaluated as dietary supplement. For the growth trials (46 and 35 days, respectively, for trials 1 and 2), the basal diet was primarily composed of soybean meal (SBM), fishmeal (FM), whole wheat, corn protein concentrate, poultry meal (PM, pet food grade) and corn starch. Non‐GM cultivars were processed with novel methodologies to produce Navita? ingredients (N1, N2 and N3) which were incorporated at low (L) or high (H) levels into the experimental diets, in partial replacement of FM and full replacement of conventional SBM. The last two formulations incorporated the fermented yeast for a total of nine experimental diets (Table 1 ). Results from the growth trials indicate that shrimp fed diet 5 (HN2) exhibited significantly lower (p < .05) weight gain as compared to shrimp fed diets 1 and 2 (basal and LN1, respectively) in trial 1, as well as compared to animals fed diets 1, 2, 3 and 8 (basal, LN1, HN1 and basal + yeast) in trial 2. The feed conversion ratio significantly increased for shrimp fed diet 5, in contrast with shrimp fed diets 1, 2, 3 and 8 in trials 1 and 2, as well as compared to shrimp fed diets diet 6 (LN3) in trial 2. For the physiological assessment (stress and immune responses), only the effects of diets 1, 3, 8 and 9 (basal, HN1, basal + yeast and HN1 + yeast, respectively) were investigated. Granular cell counts were significantly higher for shrimp fed the yeast‐containing diets. Haemolymph glucose and haemolymph packed cell volume were significantly reduced for shrimp fed diets 3, 8 and 9. No significant differences were observed in total haemocyte counts, hyaline cells counts, semi‐granular cells counts, haemolymph protein, haemocyte phagocytic capacity and haemocyte respiratory burst activity. Results of this work indicate that selective soy breeding technology coupled with novel processing options has the potential to increase the nutritional value of conventional SBM for shrimp feeds. Trends on immune responses were more difficult to elucidate possibly due to the limited length of the feeding trial.     

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller's dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 ± 0.5 °C, 32.2 ± 0.4‰) with juvenile L. vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg^− 1 CP; 8.03 g kg^− 1 lipid; 2.01 kcal g^− 1) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 °C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx²), n = 26. Best in vitro APD predictions were obtained at 25 °C, 4 U: 80 mg CP both for ingredients (R² = 0.86; P = 0.001) and test diets (R² = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD − determined APD) for corn gluten meal, feather meal, poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD > 80% showed higher prediction precision (R² = 0.98; P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R² = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information.