Evaluation of In Vitro Apparent Protein Digestibility by Shrimp Using Gut Enzyme Extracts

Knowledge of apparent protein digestibility (APD) is required for optimization of feed formulae for the production of marine penaeid shrimp. The purpose of this study was to evaluate an in vitro method for determining APD in marine penaeid shrimp using gut enzyme extracts. A high correlation (r² = 0.95) was shown between single‐ingredient APD values for fish meal diets using in vivo methodology and those derived from in vitro testing of ingredients. A second study showed positive correlation (r² = 0.71) between in vitro APD of selected purified and semipurified ingredients and their reported in vivo APDs. This correlation was much higher for purified ingredients (r² = 0.93) versus less‐refined ingredients (r² = 0.24). A third trial compared in vitro APD at three different enzyme extract pH values and showed that for most protein sources, APD was significantly highest (P < 0.05) at pH = 7.0 and lower at pH = 6.1 or 7.9, indicating a neutral pH optimum for this methodology.     

Response of Pacific White Shrimp,Litopenaeus vannamei,to Three Sources of Solvent Extracted Soybean Meal

Three sources of soybean meal (SBM) were biochemically characterized and evaluated in diets for juvenile Litopenaeus vannamei. Three biological techniques were utilized to evaluate the nutritional quality of the meals including an outdoor growth trial as well as determination of in vivo apparent digestibility and in vitro digestibility including pepsin digestibility (0.0002%), pH Stat degree of hydrolysis, and immobilized digestive enzyme assay (IDEA). The growth trial was conducted over a 10‐wk period using soybean meal as the primary protein source with each meal being included at 54–58% of the diet based on an equal protein inclusion. At the conclusion of the growth trial there were no significant differences between the test diets indicating equivalent biological availability under practical conditions. The in vivo digestibility trial utilized chromic oxide as the inert marker and 70:30 replacement techniques resulted in typical results. Apparent protein digestibility (APD) and apparent energy digestibility (AED) of the diets ranged from 66.8 to 80.2% and 65.4 to 74.7%, respectively. Ingredient digestibility values for protein and energy ranged from 78.8 to 93.5% and 60.5 to 85.0%, respectively. One of the three SBM evaluated had numerically lower protein digestibility and significantly lower energy digestibility than the other two meals .     

Prediction of culture performance of juvenile Litopenaeus vannamei by in vitro (pH-stat) degree of feed protein hydrolysis with species-specific enzymes

Rapid in vitro methods for measuring digestibility may be useful in analysing aqua feeds if the extent and limits of their application are clearly defined. The pH‐stat protein digestibility routine with shrimp hepatopancreas enzymes was previously related to apparent protein digestibility with juvenile Litopenaeus vannamei fed diets containing different protein ingredients. The potential of the method to predict culture performance of shrimp fed six commercial feeds (T3, T4, T5, T6, T7 and T8) with 350 g kg^?1 declared crude‐protein content was assessed. The consistency of results obtained using hepatopancreas enzyme extracts from either pond or clear water‐raised shrimp was further verified in terms of reproducibility and possible diet history effects upon in vitro outputs. Shrimps were previously acclimated and then maintained over 56 days (initial mean weight 3.28 g) on each diet in 500‐L tanks at 114 ind m^?2, clear water closed system with continuous renewal and mechanical filtering (50 μm), with four replicates per treatment. Feeds were offered four times daily (six days a week) delivered in trays at feeding rates ranging from 4.0% to 7.0% of stocked shrimp biomass. Feed was accessible to shrimp 4 h daily for 1‐h feeding period after which uneaten feed was recovered. Growth and survival were determined every 14 days from a sample of 16 individuals per tank. Water quality was monitored daily (pH, temperature and salinity) and managed by water back flushing filter cleaning every 7–10 days. Feeds were analysed for crude protein, gross energy, amino acids and pepsin digestibility. In vitro pH‐stat degree of protein hydrolysis (DH%) was determined for each feed using hepatopancreas enzyme extracts from experimental (clear water) or pond‐raised shrimp. Feeds resulted in significant differences in shrimp performance (P < 0.05) as seen by the differences in growth rates (0.56–0.98 g week^?1), final weight and feed conversion ratio (FCR). Shrimp performance and in vitro DH% with pond‐raised shrimp enzymes showed significant correlation (P < 0.05) for yield (R² = 0.72), growth rates (R² = 0.72–0.80) and FCR (R² = ?0.67). Other feed attributes (protein : energy ratio, amino acids, true protein, non‐protein nitrogen contents and in vitro pepsin digestibility) showed none or limited correlation with shrimp culture performance. Additional correlations were found between growth rates and methionine (R² = 0.73), FCR and histidine (R² = ?0.60), and DH% and methionine or methionine+cystine feed contents (R² = 0.67–0.92). pH‐stat assays with shrimp enzymes generated reproducible DH% results with either pond (CV ≤ 6.5%) or clear water (CV ≤ 8.5%) hepatopancreas enzyme sources. Moreover, correlations between shrimp growth rates and feed DH% were significant regardless of the enzyme origin (pond or clear water‐raised shrimp) and showed consistent R² values. Results suggest the feasibility of using standardized hepatopancreas enzyme extracts for in vitro protein digestibility.     

Effects of replacing soybean meal with rubber seed meal on digestive enzyme activity,nutrient digestibility and retention in tilapia (Oreochromis niloticus × Oreochromis aureus)

A feeding trial was conducted to evaluate the effects of replacing soybean meal (SBM) with rubber seed meal (RSM) on digestive enzyme activity, nutrient digestibility and retention in juvenile tilapia (Oreochromis niloticus × Oreochromis aureus). Five isonitrogenous and isoenergetic diets were formulated with 0 (control), 100, 200, 300 and 400 g kg⁻¹ RSM replacing graded levels of SBM respectively. Each diet was randomly assigned to triplicate groups of 30 fish (initial average weight 5.2 g) per aquarium in a rearing system maintained at 29 ± 1°C for 8 weeks. The hepatic protease and lipase activities gradually decreased with increasing dietary RSM level, but no significant differences were observed among the low inclusion level (0–200 g kg⁻¹) groups. The apparent digestibility coefficients of dry matter, crude protein, crude lipid and ash showed a similar trend as the hepatic protease and lipase activities. The retentions of protein and individual essential amino acid (except lysine, threonine and leucine) in fish fed diet with 200 g kg⁻¹ RSM were similar to those in fish fed the control diet. These results indicate that dietary RSM inclusion level up to 200 g kg⁻¹ did not markedly affect the digestive enzyme activity, nutrient digestibility and retention in tilapia, whereas these were depressed by the inclusion of 400 g kg⁻¹ RSM.     

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.     

An evaluation of replacing fish meal with fermented soybean meal in the diets of largemouth bass (Micropterus salmoides): Growth,nutrition utilization and intestinal histology

A 56‐day feeding trial was conducted to investigate the effects of replacing fish meal (FM) with soybean meal (SBM) and fermented soybean meal (FSBM) on growth performance, nutrition utilization and intestinal histology of largemouth bass. The basal diet contained 350 g/kg FM (control), and then, FM was replaced with SBM or FSBM of 15%, 30%, 45% and 60% respectively. The results showed that the specific growth rate of fish fed FSBM‐60, SBM‐45 and SBM‐60 diets significantly decreased, and the feed conversion ratio of SBM‐30, SBM‐45, SBM‐60, FSBM‐45 and FSBM‐60 groups increased when compared to the control group (p < .05). The apparent digestibility coefficients of dry matter and crude protein of SBM‐45, SBM‐60 and FSBM‐60 groups were significantly lower than those of the control group (p < .05), and the substitution of FM with SBM and FSBM (45% and 60%) significantly reduced the protein retention (p < .05). Serum total protein contents of SBM‐60 and FSBM‐60 groups and serum cholesterol contents of SBM‐45, SBM‐60 and FSBM‐60 groups were significantly lower than those of the control group (p < .05). The SBM‐30 and SBM‐60 groups showed significantly higher alanine aminotransferase activity than the control group (p < .05). The intestinal histology analysis resulted that the villus length of the SBM‐60 group and the villus width of the SBM‐45, SBM‐60 and FSBM‐60 groups decreased when compared to the control (p < .05). In conclusion, FSBM could replace 30% FM in diet of largemouth bass containing 350 g/kg FM, while the substitution level of FM with SBM was only 15%.     

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.     

Utilization of different carbohydrate sources and starch forms by bagrid catfish (Mystus nemurus) (Cuv & Val)

Two experiments were conducted to ascertain the utilization of different carbohydrate sources and different forms of starch present in bagrid catfish (Mystus nemurus) fry. The nutritional quality and starch forms of cornstarch, dextrin, broken rice, and sago flour were evaluated in a 4 × 2 factorial design. All diets were formulated to be isonitrogenous (400 g kg^?1 crude protein) and isolipidic (150 g kg^?1 crude lipid), fed to triplicate groups of bagrid catfish fry (initial weight 6.04 ± 0.04 g). Growth studies, enzymatic response and postprandial plasma glucose were observed. A two‐way anova (P > 0.05) analysis of the data obtained showed that the final body weight, specific growth rate (SGR) and food conversion ratio (FCR) were significantly affected by carbohydrate source, but not significantly affected by starch form. Fish fed broken rice and corn starch had similar SGR and FCR compared to fish fed with dextrin and sago flour. The postprandial plasma glucose trend in fish fed with corn starch and broken rice showed a gradual increase while sago flour and dextrin‐fed fish peaked at 2 and 3 h, respectively. Lipogenic enzyme (G6PDH) activity were more pronounced than gluconeogenic enzyme (ME) in M. nemurus irrespective of carbohydrate source and starch form. Therefore, we suggest that raw broken rice and corn starch are suitable carbohydrate sources in the M. nemurus diet.     

Evaluation of flash dried yeast as a nutritional supplement in plant‐based practical diets for Pacific white shrimp Litopenaeus vannamei

Growth and digestibility trials were conducted to evaluate the use of flash dried yeast (FDY) as a supplement in practical shrimp feeds. The six‐week growth trial evaluated the effects of FDY supplementation (0, 10, 20, 40 and 60 g kg^–1) in the plant‐based practical diet on the performances and proximate compositions of juvenile Pacific white shrimp, Litopenaeus vannamei. Final biomass, final mean weight, per cent weight gain (WG), feed conversion (FCR) and protein retention efficiency (PRE) of Pacific white shrimp were not significantly influenced when FDY was added up to 40 g kg^–1 of the diet. However, 60 g kg^–1 FDY supplementation significantly reduced growth, feed utilization and protein retention. Supplementation of FDY in the practical diets of Pacific white shrimp did not affect protein, moisture, lipid, crude fibre, and ash content of whole shrimp body. Apparent digestibility coefficients of energy, protein, and amino acid (AA) for FDY were determined using chromic oxide as an inert marker and the 70:30 replacement technique. The energy and protein digestibility of FDY were 38.20% and 53.47%, respectively, which were significantly lower than fishmeal (FM) and soybean meal (SBM) that were run at the same time. Most amino acid digestibility of FDY was lowest among the three ingredients tested. Results of this work indicated that energy, protein and amino acid digestibility of FDY are lower compared to SBM and FM, and high levels supplementation (≥60 g kg^–1) of FDY can cause negative effects to the growth response of Pacific white shrimp. Based on these results, further research regarding the effects of the low level inclusion of FDY in practical diets on immune responses of Pacific white shrimp is warranted.     

A comparative study on the apparent digestibility of selected feedstuffs in hybrid catfish (Clarias macrocephalus × Clarias gariepinus) and Nile tilapia (Oreochromis niloticus)

The apparent digestibility (AD) of dry matter (DM), organic matter (OM), crude protein (CP) and essential amino acids (EAA) in five selected feedstuffs was evaluated in hybrid catfish and Nile tilapia. The AD of DM, OM and CP in cassava leaf meal (CLM) was lower (P < 0.05) than in groundnut meal (GNM), soybean meal (SBM), sesame husk meal (SSH) and shrimp head meal (SHM). The AD of DM, OM and CP among GNM, SBM, SSH and SHM was not different (P > 0.05). The AD of most EAA was higher in SSH (P < 0.05) than in CLM, GNM, SBM and SHM. The AD of most EAA among CLM, GNM, SBM and SHM was not different (P > 0.05). The AD of DM and OM in CLM was higher (P < 0.05) in hybrid catfish than in Nile tilapia, while there was no difference in the AD of CP in CLM between fish species. The AD of DM, OM, CP and EAA among GNM, SBM, SSH and SHM did not differ between hybrid catfish and Nile tilapia. In conclusion, there were only small differences in the nutritional properties between the selected feedstuffs in both fish species. Most EAA in the selected feedstuffs were equally well utilized by hybrid catfish and Nile tilapia.     

Dietary supplementation of autolysed yeast enhances growth,liver functionality and intestinal morphology in African catfish

A feeding trial was conducted to evaluate the potential of dietary supplementation of autolysed brewer's yeast (AY) on African catfish. The catfish (22.5 ± 1.15 g/fish, 20 fish 33 L/tank) were fed with either of diets (390 g/kg crude protein, 140 g/kg lipid) supplemented with 0, 3, 6 or 10 g/kg AY (n = 3). After 49 days of feeding, the final body weight and metabolic growth rate of the catfish fed 3 g/kg AY (3‐AY) diet were higher than those fed the control diet (p < .05). The lowest level (p < .05) of alanine transaminase was detected in the blood of the catfish fed 3‐AY diet. The mid‐intestinal histology of the catfish revealed no significant difference (p > .05) in intestinal perimeter ratio. However, an elevated (p < .05) abundance of goblet cells and intraepithelial leucocytes were found in the intestine of catfish fed 3, 6 and 10 g/kg AY diets, with the highest level of abundance recorded in the mid‐intestine of the catfish fed 3‐AY diet. The results suggest that dietary 3 g/kg autolysed brewer's yeast supplementation improves growth performance of African catfish without deleterious effect on liver functionality and gut morphology.     

Comparative Characterization of Enzymatic Digestion from Fish and Soybean Meal from Simulated Digestive Process of Pacific Bluefin Tuna,Thunnus orientalis

The digestive process of the Pacific bluefin tuna (PBT), Thunnus orientalis, was simulated through two phases of in vitro digestion: acidic digestion with porcine pepsin, followed by alkaline digestion with pancreatic crude extract (PCE) obtained from the PBT to hydrolyze fish meal (FM) and soybean meal (SBM) as protein substrates. The crude protein from FM resulted in a lower degree of hydrolysis (73.3%) compared with SBM (79.2%). However, the resulting digested products showed that FM contained 35% more small peptides, with sizes <6.5 kDa than those from the starting material (>150 kDa). The SBM had an increase of only 1.3% in the similar peptide cut‐offs found after hydrolysis. These results suggested that FM appeared to be a better source of protein according to the amount of low‐molecular weight peptides. In addition, the proteolytic activity of PCE showed that 88.9% of its alkaline proteolytic activity corresponded to trypsin and 2.9% corresponded to chymotrypsin activity. The results shown here demonstrate that peptide sizes are important in identifying suitable protein sources for aquafeed production to reinforce the primary results obtained from the in vitro digestibility using the pH‐Stat system. These results also contribute to a better understanding of the digestibility process in aquatic organisms.     

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.     

Induction of meiotic gynogenesis in bagrid catfish (Pseudobagrus ussuriensis) with homologous sperm and its confirmation for female homogamety

The bagrid catfish, Pseudobagrus ussuriensis, exhibits significant sexual dimorphism in growth rate and body size with males growing faster than females. Therefore, an all‐male culture can dramatically increase the output and profitability of fishery products. According to the monosex breeding route, super‐male individuals’ acquirement by XY male sex reversal and artificial gynogenesis is the key step. An effective protocol to induce meiotic gynogenesis using homologous sperms has been developed in this study. The most effective UV irradiation for sperm genetic inactivation was found to be at a distance of 20 cm with 66 μW/cm² light intensity for 25 min. Optimal treatment for cold shock was 5 min post‐fertilization at 0‐4°C for 30 min, which gave the best survival rate of 13.65 ± 2.87%. The sex ratio in the meiotic gynogens showed a significant female‐biased deviation (p < .05); thirty meiogynogens and their parents were further analysed using a male‐specific AFLP marker, of which only the male parent produced a male‐specific DNA band of 412 bp. These results indicated the female homogametic XX/XY sex determination system in P. ussuriensis. The optimization of a protocol for the successful induction of meiogynogenesis in the bagrid catfish lays the basis for all‐male production and is useful in ascertaining the genetic sex determination system in this promising aquaculture species.     

Comparative effects of dietary supplementation with maggot meal and soybean meal in gibel carp (Carassius auratus gibelio) and darkbarbel catfish (Pelteobagrus vachelli): growth performance and antioxidant responses

A 6‐week growth trial was conducted to investigate the effect of dietary supplementation with maggot meal (MGM) and soybean meal (SBM) on the growth performance and antioxidant responses of gibel carp (GC) and darkbarbel catfish (DC). The basal diet was formulated to contain 114 g kg⁻¹ fish meal (FM) and 200 g kg⁻¹ SBM. The basal diet was supplemented with either 280 g kg⁻¹ FM (Control), 390 g kg⁻¹ MGM or 450 g kg⁻¹ SBM to obtain three isonitrogenous (crude protein: 380 g kg⁻¹) and isocaloric (gross energy: 16 kJ g⁻¹) diets. For GC, a significant decrease in specific growth rate (SGR) was only observed in fish fed the SBM diet compared with the control (P < 0.05). Principal components analysis (PCA) of GC showed a higher similarity in antioxidant response to dietary supplementation with MGM and SBM proteins between liver and intestine, but the DC did not. The present results suggest that supplementing 390 g kg⁻¹ MGM protein to basal diet cause an enhancement of the antioxidant capacity in GC, but supplementing 390 g kg⁻¹ MGM and 450 g kg⁻¹ SBM proteins to basal diets resulted in a significant attenuation of the antioxidant capacity in DC.     

Utilization of combined extruded soybean and corn gluten meals as feed ingredients for juvenile rainbow trout,Oncorhynchus mykiss diet

In this study, feeding experiment and subsequent digestibility trial were performed to investigate the utilization of extruded soybean meal (SBM) and corn gluten meal (CGM) as feed ingredients for juvenile rainbow trout. Plant ingredients have undergone extrusion at low temperature (100°C, LT) or high temperature (150°C, HT) for 30 s. Four isonitrogenous (44%, crude protein) and isolipidic (14%, crude lipid) diets were formulated. Control diet is fishmeal based while a combined (1:1) non‐extruded SBM and CGM for NE diet, LT SBM and LT CGM for LT diet and HT SBM and HT CGM for HT diet. Two hundred forty rainbow trout juveniles (7.8 g average body weight) were randomly divided into 12 rectangular 60 L glass aquaria and offered four different diets in triplicate. Fish were fed at satiation twice a day, six days a week for 12 weeks. Phosphorus in phytic acid level of extruded ingredients decreased through extrusion cooking. Final weight, weight gain, SGR and PER of fish fed HT diet were significantly (p < .05) higher than those fed with NE diet. Apparent digestibility coefficient for protein of LT and HT diets is significantly higher than of NE diet. The results of this study demonstrated that HT extruded SBM and CGM are suitable feed ingredients for rainbow trout diet without compromising fish growth, feed utilization and fish body composition.     

Amino acid availability and protein digestibility of several protein sources for Nile tilapia,Oreochromis niloticus

Apparent amino acid availability coefficients and protein digestibility of four animal products [fish meal (FM), meat and bone meal (MBM), poultry by‐product and feather meal] and four plant protein‐rich products [soybean meal (SBM), cottonseed meal‐28, cottonseed meal‐38 and corn gluten meal (CGM)] were determined for Nile tilapia, Oreochromis niloticus. Ingredients were incorporated to a practical reference diet at a 7 : 3 ratio (70% of reference diet and 30% of test ingredient). Chromic oxide was used as external digestibility marker. Among animal products poultry by‐product meal (PBM; 89.7%) and FM (88.6%) presented the highest apparent protein digestibility (APD) while MBM (78.4%) and feather meal (78.5%) presented the lowest APD. Among plant protein‐rich products CGM (91.4%) and SBM (92.4%) presented the highest APD values while cottonseed meal‐28 presented the lowest APD (78.6%). Average apparent amino acid availability of feed ingredients was similar to protein digestibility with 92.3%, 89.6%, 73.4%, 80.7%, 88.9%, 84.4%, 91.2% and 79.7% values for SBM, CGM, cottonseed meal‐28 and 38, FM, MBM, PBM and feather meal respectively. These results indicate that O. niloticus is able to utilize efficiently different feedstuffs.     

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.     

Apparent energy,dry matter and amino acid digestibility of differently sourced soybean meal fed to Pacific white shrimp Litopenaeus vannamei

Due to the variations in nutrient quality of soybean meal (SBM) that is a result of differences in production location and processing specifications, a study was conducted to determine the fluctuations in apparent digestibility coefficients of differently sourced SBM fed to Pacific white shrimps (Litopenaeus vannamei). Twenty‐four SBM‐based diets were formulated by mixing a basal diet and test ingredients on a dry matter basis (70:30 ratio), while 1% chromic oxide was used as the inert marker. The digestibility trial was carried out in a semi‐closed recirculation system with six replicate groups per treatment (mean shrimp weight of 10.2 g). Significant differences were observed for apparent dry matter, energy and protein digestibility coefficients (p < .05 was considered significant) among 24 sources of SBM and digestibility values ranged from 45% to 90%, 56% to 93% and 87% to 98%, respectively. Based on multivariate analysis, acid detergent fibre, neutral detergent fibre, lignin, raffinose and trypsin inhibitor were screened as the key chemical characteristics in SBM that influenced digestibility of nutrients in Pacific white shrimps. Variations in growth performances of shrimp were in line with the variations in apparent digestibility coefficients of SBM verifying the importance of digestibility data in shrimp feed formulations.     

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.