Digestive capacity, intestinal morphology, and microflora of 1-year and 2-year old Atlantic cod (Gadus morhua) fed standard or bioprocessed soybean meal

The objectives of this work were to evaluate how dietary soybean meal (SBM) or a soy meal made by bioprocessing the SBM (BPSBM) to remove anti-nutritional factors affected hydrolytic capacity, amino acid absorption, intestinal morphology, and microflora along the intestinal tract of Atlantic cod at two life stages. Three fish meal based standard cod diets were formulated to contain no soy (FM control), 25% SBM, or 22% BPSBM. Prior to sampling the diets were fed to duplicate groups of 0.5 kg (1-year old) and 1.7 kg (2-year old) cod for a period of 3 months, and the groups reached 0.9 and 2.5 kg, respectively. Digesta was then sampled from different intestinal sections for analyses of trypsin and amylase activity as well as absorption of amino acids, nitrogen, and sulphur. Gastrointestinal sections were sampled for measurements of relative weight (kg^− 1 body weight), and tissues from these sections were sampled for analyses of brush border enzyme (alkaline phosphatase (ALP), leucine aminopeptidase (LAP), and maltase) activity and histological examination. Microflora was sampled from both digesta and the intestinal wall. The SBM diet stimulated relative growth of all gastrointestinal sections except the distal intestine in both age classes. Relative growth of the pyloric intestine was also stimulated by BPSBM. The pyloric caeca and the upper mid intestine were found to be the major sites for enzymatic hydrolysis of protein and starch and for amino acid absorption. Dietary SBM and BPSBM did not alter the activity of trypsin and LAP, but the activity of these enzymes in the proximal intestine was affected by age, being higher in 1-year old than in 2-year old cod. The rate of amino acid, nitrogen, and sulphur absorption along the gastrointestinal tract was not affected by SBM, but was slowed by BPSBM. Dietary SBM or BPSBM did not alter the morphology of the intestinal mucosa in any sections of the cod intestine. The distal-most structure of the intestine, a compartment with inlet and outlet (anus) valves, showed very high microbial colonisation in the mucosal brush border. Inclusion of SBM in the diet changed the intestinal microflora, increasing the population level of transient bacteria in the pyloric and mid intestine, but reducing the population level of adherent bacteria throughout the intestine. To conclude, Atlantic cod appeared to have a robust and flexible digestive system able to adjust to high dietary levels of soy protein meals.     

Digestive function and intestinal integrity in Atlantic salmon (Salmo salar) fed kernel meals and protein concentrates made from yellow or narrow-leafed lupins

This study assessed the effects of yellow lupin (Lupinus luteus) and narrow-leafed lupin (L. angustifolius) kernel meals and protein concentrates on the gastrointestinal integrity, capacity for digestive hydrolysis, and digestibility of nutrients in Atlantic salmon. A basal diet (FM) was made from fish meal, wheat, and fish oil. Six additional diets were formulated by replacing 30% of the FM diet with lupin kernel meal made from L. l. cv. Wodjil (LKM), L. a. cv. Belara (BKM), and L. a. cv. Myallie (MKM), lupin protein concentrates made from the same L. l. (LPC) and L. a. cv. M (MPC), or extracted soybean meal (SBM). All diets were extruded. Each diet was fed to three groups of 176 g salmon kept in 1 m² tanks with 5.6 °C saltwater for 3 weeks prior to sampling of blood, intestinal organs, digesta, and faeces. Inclusion of lupin meals in the diets resulted in harder and more condensed feed particles. Ulcer-like lesions were observed in the stomach of fish from all feeding groups, and this was worsened by lupin in the diet, but did not appear to be pellet hardness related. No consistent altered morphology was observed in the distal intestine (DI) of fish fed the FM and lupin diets, while the DI of fish fed SBM showed consistent and typical soybean meal-induced pathomorphological changes. Plasma cholesterol was higher when feeding MKM and LKM than when feeding FM, MPC, and LPC, with intermediate levels when feeding BKM and SBM. Feeding LKM and LPC resulted in a higher weight of the GIT when related to body weight. Trypsin activity and bile acid concentration were generally higher in digesta from the pyloric (PI) and mid (MI) intestine when feeding FM and lupin diets than when feeding SBM, while the opposite was seen for trypsin activity in digesta from DI. There were no effects of diet on leucine aminopeptidase (LAP) and maltase activity in PI and MI, but in DI the activity of these brush border enzymes were significantly lowered when feeding SBM. SBM in the diet resulted in watery faeces and lowered apparent digestibility of lipid, but this was not observed when feeding the lupin diets. To conclude, the tested lupin kernel meals and protein concentrates did not alter the intestinal function in Atlantic salmon when included at 30% of the diet. Dietary lupin was, however, involved in the worsening of ulcer-like gastric lesions.     

Effects of graded levels of standard soybean meal on intestinal structure, mucosal enzyme activities, and pancreatic response in Atlantic salmon (Salmo salar L.)

Duplicate tanks of c.280 g Atlantic salmon (Salmo salar L.) were fed for 60 days on diets in which fishmeal was substituted with graded levels of extracted soybean meal (SBM) comprising 0%, 10%, 15%, 20%, 25% or 35% of total protein. The effects on feed intake, growth, feed conversion, apparent digestibility and utilization of macronutrients and energy, pathohistological response of the distal intestine (DI), activities of digestive enzymes in the mid and distal intestinal mucosa, and faecal trypsin and plasma insulin concentrations were studied. A negative, dose‐dependent effect of SBM was observed on nearly all performance parameters with a notable exception of feed intake. The lowest SBM inclusion level of 10% resulted in moderate pathohistological changes in the DI. Each subsequent increase in SBM level increased the number of fish displaying severe changes. In contrast to the mid‐intestine (MI), all enzyme activities in the distal intestinal mucosa decreased dose‐dependently with increasing SBM inclusion. Faecal trypsin increased up to an SBM inclusion level of 20% and then levelled off. Plasma insulin increased from 0% to 15% SBM inclusion and then decreased. The results suggest that caution should be exercised in the use of even low levels of extracted SBM in salmon feeds.     

Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

Changes in immune and enzyme histochemical phenotypes of cells in the intestinal mucosa of Atlantic salmon,Salmo salar L., with soybean meal‐induced enteritis

Extracted soybean meal (SBM) in the diet for Atlantic salmon, Salmo salar L., causes an inflammatory response in the distal intestine. The morphological changes of the epithelial cells and a characterization of the inflammatory cell infiltrate of the distal intestinal mucosa were studied using a panel of enzyme and immunohistochemical markers. The salmon (average body weight 927 g) used in the study were fed either a fishmeal‐based diet (control diet) or a diet in which 30% of the fishmeal protein was replaced with SBM protein (SBM diet). In salmon fed SBM, there were markedly reduced enzyme reactivities in the distal intestinal epithelial cells, both in the brush border [5′‐nucleotidase (5′N), Mg2+‐ATPase, alkaline phosphatase (ALP) and leucine aminopeptidase (LAP)] and in the intracellular structures [alkaline and acid phosphatase, non‐specific esterase (NSE) and alanine aminopeptidase (AAP)]. There appeared to be an increased presence of cells of monocytic lineage, including macrophages, as well as neutrophilic granulocytes and immunoglobulin (Ig) M in the lamina propria of the SBM‐fed fish. The mid intestine showed little response to the diet. The results suggest that toxic/antigenic component(s) of SBM affect the differentiation of the distal intestinal epithelial cells and may help explain the reduced nutrient digestibilities previously reported in salmonids fed extracted SBM.     

Nutrient digestibilities and gut function in Atlantic salmon (Salmo salar) fed diets with cellulose or non-starch polysaccharides from soy

Duplicate groups of Atlantic salmon (Salmo salar), kept in saltwater, were fed fish meal based cold-pelleted diets where graded levels of native or extruded non-starch polysaccharides (NSP) from soybeans replaced cellulose, at a total NSP level of 100 g kg^− 1 for 28 days. The study also included a diet where defatted soybean meal (SBM) constituted the NSP at a level of 100 g kg^− 1 and a reference diet without NSP supplementation.The SBM diet resulted in a significant reduction in faecal dry matter content, apparent digestibilities of starch and organic material, and growth, and an increased faecal output of several elements (K, Na, Zn), compared to all the other diets. Morphological changes were only seen in the intestine of the fish fed the SBM, i.e. the diets holding purified soy-NSP did not induce enteritis. When compared to the diet without NSP, cellulose addition increased faecal dry matter, while inclusion of native soy-NSP reduced it. Dry matter in faeces and apparent digestibility of crude protein decreased in a linear manner, while the digestibility of starch and faecal output of K and Na increased linearly when native soy-NSP replaced graded levels of cellulose from 0 to 100 g kg^− 1 feed. When diets with 75 or 100 g kg^− 1 of native NSP and extruded soy-NSP were compared, fish fed native soy-NSP had reduced faecal dry matter, higher digestibility of starch, and increased faecal output of Cu, Fe, and K. Dry matter in faeces and faecal output of Cu was lower for the highest inclusion level, while digestibility of starch and faecal output of Mn and K were higher.In conclusion, soy-NSP was inert compared to the fish meal reference, with respect to nutrient digestibilities and intestinal pathologies, but affected faecal mineral excretion in Atlantic salmon.     

Pacific hake (Merluccius productus Ayres, 1855) hydrolysates as feed attractants for juvenile Chinook salmon (Oncorhynchus tshawytscha Walbaum, 1792)

Soybean meal (SBM) inclusion in salmonid diets can lower feed cost, but dramatically reduces growth and feed utilization, and increases mortality in juvenile chinook salmon Oncorhynchus tshawytscha, due to diminished diet palatability and/or other adverse physiological effects exerted by antinutritional factors in SBM. The objective of this study was to investigate whether commercial Antarctic krill meal Euphausia superba or hydrolysates enzymatically produced from Pacific hake Merluccius productus could reverse the negative palatability effects of SBM inclusion in juvenile chinook salmon diets. Diets without SBM or with SBM and no added feed attractant were used as positive and negative control diets respectively. Incorporation of 2% krill meal or Alcalase^®‐produced hydrolysates into SBM‐containing diets (20% of dry matter by isonitrogenous replacement of fishmeal) significantly (P < 0.05) increased feed intake, feed utilization, fish weight gain and thermal growth coefficient during a 5‐week trial. Nevertheless, the negative effects on fish performance incurred by dietary inclusion of 20% SBM could not be fully reversed, indicating that most of those effects were likely unrelated to palatability. This study demonstrates the potential for using Pacific hake hydrolysates as a dietary feed attractant for salmonid diets, and supports the need for further research to optimize its application for ideal fish performance.     

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.     

Effects of dietary sodium butyrate on growth,diet conversion,body chemical compositions and distal intestinal health in yellow drum (Nibea albiflora,Richardson)

The present study was conducted to investigate the effects of sodium butyrate (SB) on the physical barrier function, pro‐inflammatory response and possible underlying mechanisms in the distal intestine (DI) of yellow drum when fed a high‐SBM diet. Three iso‐proteic and iso‐lipidic diets were formulated with fish meal (FM, the control), 45% fish meal protein replaced by SBM (SBM) and 45% fish meal protein replaced by SBM but supplemented with 0.15% SB (SBM + SB). Fish were fed twice daily for 10 weeks. Results indicated that SB supplementation significantly increased the specific growth rate (SGR) and feed efficiency ratio (FER) and methionine content of muscle when compared with those of fish fed the SBM diet (p < .05). The morphologic histology results showed that SB dramatically improved the physical barrier structure, characterized as increases of fold height (FH) and muscular thickness (MT) (p < .05). RT‐qPCR data were accordant with morphologic histology results, in which claudin 3, claudin 4 and occludin were increased while claudin 7 and myosin light chain kinase (MLCK) mRNA expression levels were decreased (p < .05). Sodium butyrate also exerted anti‐inflammatory function, which may be attributed to the suppression of nucleus p65 protein expression. Results suggest SB can be incorporated in high‐SBM diets to ameliorate the negative consequences of alternative dietary ingredients on yellow drum physiology.     

Evaluation of the effect of commercially available plant and animal protein sources in diets for Atlantic salmon (Salmo salar L.): digestive and metabolic investigations

The present study investigated the effect of various alternative diet ingredients partially replacing fishmeal (FM) on digestive and metabolic parameters in Atlantic salmon (Salmo salar L.) post-smolts (initial body mass 305 ± 69 g) following 12 weeks of feeding. Experimental diets containing 20 % extracted sunflower (ESF), pea protein concentrate (PPC), soy protein concentrate (SPC), feather meal (FeM) and poultry by-product (PBY) were compared to a reference diet containing FM as the main protein source. For the different intestinal compartments trypsin, lipase, bile salts, dry matter and chyme-associated leucine aminopeptidase (LAP) were measured from the content and LAP was measured in the tissue. Selected metabolites were measured in plasma samples. In general, use of plant proteins resulted in low C-LAP activity, low plasma cholesterol and high plasma magnesium. The plasma levels of cholesterol and Mg reflecting were most likely reflections of the composition of the diet, while the LAP activity in chyme may indicate lower epithelial cell turnover. Other responses varied depending on the plant protein source. Results from the animal protein substitution also varied both between diets and compartments; however, both materials increased lipase activity in DI. FeM resulted in a significant increase in both total and specific LAP activities suggesting an attempt to increase the digestive capacity in response to low digestibility of the diet while PBY showed very little difference from the FM-fed control fish. The present trial indicates that 20 % PPC, SPC and PBY can partially replace FM in diets for Atlantic salmon. The qualities of ESF and FeM used in this trial show little promise as FM replacement at 20 % inclusion level.     

Fishmeal can be totally replaced by a mixture of rapeseed meal and Chlorella meal in diets for crucian carp (Carassius auratus gibelio)

This study was conducted to investigate the effects of dietary fishmeal (FM) replacement by a mixture of rapeseed meal and Chlorella meal (RCM) on growth performance, apparent digestibility coefficients (ADCs), digestive enzymatic activities and intestinal histology of crucian carp Carassius auratus gibelioi. Five isonitrogenous diets were formulated to replace 0% (RCM0), 25% (RCM25), 50% (RCM50), 75% (RCM75), and 100% (RCM100) of protein from fishmeal with RCM respectively. Each experimental diet was randomly assigned to triplicate groups with 25 juvenile fish (initial body weight: 1.77 ± 0.04 g) per fibreglass tank for 6 weeks. With increasing substitution levels, weight gain rate, specific growth rate, feed intake and protein efficiency ratio increased, but feed conversion rate decreased. Dietary RCM substitution improved lipid content of muscle, but had no significant effect on other proximate composition of muscle and liver. ADCs of dry matter, protein, lipid, energy and the majority of amino acids increased with increasing substitution level, and digestive enzyme activities (amylase, trypsin and lipase) in intestine showed the similar trend with ADCs. Dietary RCM substitution had no significant adverse effect on intestinal histology. This study indicated that FM protein could be completely replaced by mixed protein sources (RCM) in crucian carp diets.     

The effects of dietary inulin on growth performances,survival and digestive enzyme activities of common carp (Cyprinus carpio) fry

This study was conducted to investigate the effects of dietary inulin on growth performance, diet utilization, survival rate, carcass composition and digestive enzymes activities (amylase, lipase and protease) of carp (Cyprinus carpio) fry (0.55 ± 0.02 g). After acclimation, fish were allocated into 9 tanks (40 fish per tank) and triplicate fish groups were fed, control diet (0 g) or diets containing 5 g and 10 g inulin kg^?1 for 7 weeks. No significant effect on growth performance and diet utilization of fish fed inulin compared with the control group was observed. However, supplementation of inulin significantly increased survival rate and carcass lipid content, while carcass protein content significantly decreased. Dietary inulin had no significant effects on digestive lipase, protease and amylase activities.     

Effects of fructooligosaccharide on growth,immunity and intestinal microbiota of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

The effects of fructooligosaccharide (FOS) on growth performance, immunity and predominant autochthonous intestinal microbiota of shrimp (Litopenaeus vannamei) fed diets with fish meal (FM) partially replaced by soybean meal (SBM) were evaluated. After acclimation, shrimps (1.82 ± 0.01 g/kg) were allocated into 15 tanks (25 shrimps per tank) and fed five different diets including positive control diet (C0, containing 250 g/kg FM and 285 g/kg SBM), control diet (C, containing 125 g/kg FM, 439 g/kg SBM) and three experimental diets supplemented with 1.0 g/kg FOS (T1), 2.0 g/kg FOS (T2) and 4.0 g/kg FOS (T3) to control diet (C) respectively. Shrimps were fed diets to apparent satiation three times per day, and 15 shrimps from each aquarium were randomly sampled and analysed at the end of the 6‐week feeding trial. The results showed that FBW, WGR, SGR and SR decreased, while FCR and FI increased significantly in control (C) compared with positive control (C0). Besides, significantly decreased trypsase and lipase activities, and SOD, AKP and ACP activities were recorded in control (C) compared with positive control (C0). On the other hand, significantly improved SGR and decreased FCR were observed in groups T1, T2 and T3 compared with control (C). Moreover, lipase and amylase activities enhanced significantly in group T3 compared with the control (C), while GOT and GPT activities dropped significantly with the increment supplementation of FOS in diets. Compared with the control (C), SOD activity enhanced significantly and MDA level decreased significantly in groups T2 and T3, and improved AKP and ACP activities were observed in group T3. In addition, dietary FOS improved the microbial diversity, and suppressed several potential pathogens, such as Vibrio tubiashii, Vibrio parahaemolyticus and Photobacterium damselae‐like strains in the intestine of shrimp. Overall, these results proved FOS could relieve the side effects induced by SBM and supported the use of 2.0–4.0 g/kg FOS in shrimp diets with FM partially replaced by SBM.     

Effect of replacing fish meal with soybean meal on growth, feed utilization and carcass composition of cuneate drum (Nibea miichthioides)

The effect of replacing fish meal with soybean meal (SBM) in practical feeds for cuneate drum was evaluated in an 8-week net pen trial. The cuneate drum fingerlings (initial body weight 29.8 ± 1.3 g fish^− 1) were fed six isonitrogenous and isocaloric feeds containing 39% digestible protein and 16 MJ kg^− 1 digestible energy. The control feed was formulated to contain 40% herring meal, whereas in the other five feeds SBM was included at 11.3, 22.5, 33.8, 45.0 and 56.3% to replace 20, 40, 60, 80 or 100% of the fish meal. There were no significant differences in feed intake between fish fed the control feed and feeds in which SBM replaced 20 to 80% of the fish meal, but fish fed the fish meal free feed had higher feed intake than the other treatments. Weight gain linearly declined with the decrease of fish meal level. Final body weight (FBW) of fish fed the feeds in which SBM replaced 20% of the fish meal did not significantly differ from fish fed the control feed. Replacing 40 to 100% of the fish meal resulted in lower FBW and nitrogen retention efficiency (NRE), and higher feed conversion ratio (FCR) than those of fish fed the control feed. Fish fed the feeds in which SBM replaced 60 to 100% of the fish meal had lower condition factor and hepatosomatic index than those of fish fed the control feed. No significant differences in carcass protein content was found among the treatments, but fish fed the feeds in which SBM replaced 60 to 100% of the fish meal had higher moisture and lower lipid content in carcass than those of fish fed the control feed. Results of the present study appear to indicate that cuneate drum has a limited ability to utilize SBM as a protein source in practical feeds.     

Influence of poultry meal, meat meal or soybean meal inclusion on weight gain and production characteristics of Australian snapper Pagrus auratus

Two experiments were done to evaluate the effects of poultry meal (PM), meat meal (MM) or solvent-extracted soybean meal (SBM) inclusion on the performance of Australian snapper Pagrus auratus. In each experiment, test feeds were formulated with similar contents of digestible protein (DP) and digestible energy (DE) using previously determined digestibility coefficients for this species. In experiment 1, groups of snapper (initial weight 14 g) were fed 4 feeds containing 360, 480, 610 or 730 g kg⁻¹ PM; 3 feeds containing 345, 320 or 500 g kg⁻¹ MM; 3 feeds containing 420, 600 or 780 g kg⁻¹ SBM. In experiment 2, groups of snapper (initial weight 87 g) were fed 3 extruded test feeds that contained combinations of PM, MM, SBM or blood meal (BM) which replaced all but 600, 250 or 160 g kg⁻¹ of fishmeal in respective diet formulations. Both experiments included a proprietary extruded aquafeed (COM) to benchmark fish performance. In experiment 1, weight gain was highest in snapper fed feeds containing 360, 345 or 420 g kg⁻¹ of PM, MM or SBM, respectively, and was similar (P > 0.05) to snapper fed the COM feed. Nonetheless, weight gain and protein retention efficiency tended to decrease as the amount of each test ingredient was increased. Relative feed intake was not affected by the inclusion level of PM, MM or BM, but declined significantly in snapper fed diets containing 600 or 780 g kg⁻¹ SBM. Feeding behaviour indicated fish found these feeds unpalatable. In experiment 2, the harvest weight of snapper fed the 3 extruded test feeds was similar (P > 0.05), but lower than snapper fed the COM feed (i.e. 234 vs. 256 g). Feed conversion ratio (FCR) was best in snapper fed the COM feed (FCR = 1.53); however, the FCR of snapper fed feeds containing 160 (FCR = 1.66), 250 (FCR = 1.70) or 600 g kg⁻¹ fishmeal (FCR = 1.60) was not different (P > 0.05). Australian snapper will readily accept feeds containing high levels of PM, MM or SBM and feeds containing these ingredients will support rapid weight and protein gain with little affect on whole body composition. In combination, these feed ingredients were able to replace all but 160 g kg⁻¹ of fishmeal in an extruded test feed. As such, they serve as valuable alternatives to fishmeal and extend the manufacturing options available to aquafeed producers.     

Ontogenic changes in the digestive enzyme activities and the effect of different starvation duration on the digestive enzyme activities of larval red swamp crayfish (Procambarus clarkii)

This study was performed to determine the effect of starvation and delayed feeding on activities of digestive enzymes and alkaline phosphatase (ALP) of larval red swamp crayfish (Procambarus clarkii), so as to reveal the tolerance to prolonged starvation and the recovery of digestive enzymes after delayed feeding in larval and juvenile P. clarkii. In the control group, activities of trypsin and ALP increased significantly (p < .05) with day‐age and then kept constant at 24 days after hatching (DAH) and 10 DAH, respectively, whereas the activities of amylase and pepsin increased firstly then decreased with day‐age, and the activity of lipase increased firstly then decreased and then increased again during the development period of juvenile P. clarkii (1–31 DAH). In the group with continuous starvation (CS), activities of pepsin and lipase both decreased (p < .05) after fasting, and the activities of pepsin, lipase and trypsin in the groups with delayed feeding all increased (p < .05) and recover to the levels of the control group after food supply. However, the activity of amylase increased (p < .05) in the CS group, and it decreased to normal level after food supply. The ALP activity did not significantly (p > .05) vary after starvation, whereas it decreased in the groups with delayed feeding after 1 day of food supply, and then increased back to the level similar with the control group. Results from this study could provide information for diet preparation and feeding regime in larval and juvenile red swamp crayfish culture.     

Effects of fish origin probiotics on growth performance,immune response and intestinal health of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

This study was conducted to evaluate the application of four fish origin probiotics to relieve the side effects induced by SBM in shrimp. Shrimps were fed with high fish meal diet (C0, positive control containing 500 g/kg FM and 0 g/kg SBM), low fish meal diet (C, control containing 250 g/kg FM and 340 g/kg SBM) or the control diet (C) supplemented, respectively, with 1.0 × 10⁸ CFU/g Bacillus pumilus SE5 (T1), Psychrobacter sp. SE6 (T2), Enterococcus faecium MM4 (T3) or Bacillus claussi DE5 (T4). After 6 weeks of feeding, the growth performance, hepatopancreatic digestive enzymes, intestinal histological structure and immune parameters were determined. The results showed that compared with high FM group (C0), significantly lower WGR and SGR as well as higher FCR were observed in the low FM group (C) (p < .05). Compared with the control, significantly lower FCR was observed in treatment T1 (p < .05), but not in treatments T2–T4. Protease, amylase and lipase activities in all the treatments (T1‐T4) were significantly higher than the control (p < .05). GOT and GPT activities in the control were significantly lower than the C0 (p < .05), while higher MDA level was recorded in the control (p < .05). Meanwhile, higher GPT activities were observed in treatments T1‐T4 compared with the control (p < .05). Compared with the C0, lower SOD, ACP and AKP activities were observed in the control. Significantly improved SOD and AKP activities were observed in all probiotic feeding groups compared with the control (p < .05). The control diet led to significant reduction of intestinal wall thickness and villus height compared with the C0 (p < .05), while significantly higher intestinal wall thickness and villus height were exhibited in all the probiotic groups compared with the control (p < .05), except intestinal wall thickness in treatment T3. Thus, these results demonstrated that the four fish origin probiotic strains could relieve the side effects induced by high level of SBM in shrimp, while probiotic B. pumilus SE5 showed the best performance.     

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.     

Replacement of soya bean meal with cashew nut meal as an alternative protein source in the diet of tilapia,Oreochromis mossambicus

An experiment was conducted to replace soya bean meal (SBM) with cashew nut meal (CNM) in the diet of tilapia (Oreochromis mossambicus). Five isonitrogenous (30% CP) and isolipidic (6% CL) diets, CNM0 to CNM4, were prepared replacing SBM at 0%, 12.5%, 25%, 37.5% and 50% with CNM. The feeding trial was conducted in FRP digestibility tanks of 150 L capacity for 60 days. Twenty tilapia fry (20.56 ± 0.05 g) were stocked in each tank. The final weight, weight gain and weight gain% of tilapia that received the CNM4 diet were significantly higher (p < .01) among the treatments. The significantly higher nutrient utilization efficiency in terms of FCR, PER, PRE and LRE was obtained in the CNM4 group. Similarly, a significantly lower (p < .01) body moisture content and a significantly higher (p < .05) body protein content were also observed in the CNM4 diet‐fed group. In tilapia fed the CNM4 diet, the activity of both protease and amylase enzymes was significantly higher (p < .05), but the amylase:protease ratio (A:P) was significantly lower (p < .05). Protease, amylase and A:P were positively correlated with the nutrient digestibility coefficient. The serum protein, albumin and globulin of tilapia fed the CNM4 diet were significantly higher (p < .05) and the liver status enzymes did not vary (p > .05) among the treatments. It can be concluded from the study that 50% replacement of SBM with CNM in the diet is suitable for the enhancement of growth, nutrient utilization and health status of tilapia.     

Using Bacillus subtilis E20‐fermented soybean meal as replacement for fish meal in the diet of orange‐spotted grouper (Epinephelus coioides,Hamilton)

The potential of Bacillus subtilis E20‐fermented soybean meal (FSBM) as a partial alternative component of fish meal (FM) in fed diets of orange‐spotted grouper (Epinephelus coioides) was evaluated in this study. An FM‐based diet and seven diets containing 10%, 20% and 30% and 10%, 20%, 30% and 40% of FM replaced by soybean meal (SBM) and FSBM, respectively, were fed to grouper for 84 days to evaluate possible substitution levels of FM by tracking growth performance, digestive enzyme activity and morphological changes in the liver and distal intestine. No significant differences in survival and muscle composition of grouper were found between controls and treatments. Growth performance and feed efficiency of fish fed diets with FM replaced by FSBM up to 30% were not significantly different from controls, whereas significantly decreased growth performance and feed efficiency occurred with diets containing >20% of SBM. Based on the feed efficiency, the maximum substituted levels of FM by SBM and FSBM in grouper diets were 18.36% and 29.32%, respectively, based on broken‐line analyses. Histopathological changes in the liver and distal intestine, and significantly lower activity levels of digestive enzymes, including pepsin in the stomach and trypsin, chymotrypsin, amylase and lipase in the distal intestine, were found in fish fed a diet containing 30% of FM replaced by SBM. However, these parameters were improved by the substitution of FSBM. It is therefore believed that FSBM has great potential to be used as a protein source in grouper diets in partial replacement of FM.