Effects of synbiotic containing Lactobacillus plantarum 7–40 and galactooligosaccharide on the growth performance of white shrimp,Litopenaeus vannamei

This study aimed to develop a synbiotic combination with probiotic, Lactobacillus plantarum 7–40 and one of three prebiotics, fructooligosaccharide (FOS), galactooligosaccharide (GOS) and mannan oligosaccharide (MOS). The best in vitro growth was observed when probiotic was cultured in the medium containing either FOS or GOS as the sole of carbon source. The analysis of enzyme activity revealed that GOS induced the highest activities of protease and β‐galactosidase of probiotic. Based on the findings, probiotic + GOS were selected as synbiotic to evaluate if it could promote the growth of white shrimp, Litopenaeus vannamei. For this, four diets, including a basal diet with no GOS or probiotic (control), 0.4% GOS (PRE), 10⁸ CFU probiotic kg^?1 (PRO) and 0.4% GOS in combination with 10⁸ CFU probiotic kg^?1 (SYN), were fed to shrimp for 60 days, and then the growth performance, intestinal microbiota (including total Vibrio counts, VBCs; and lactic acid bacteria, LAB) and digestive enzyme (including protease, leu‐aminopeptidase and β‐galactosidase) were evaluated. The weight gain (WG) of shrimp fed the PRO did not significantly differ from those of control (p > .05). Shrimp fed the SYN had significantly higher WG compared with the other treatments (p < .05). In addition, the SYN‐fed shrimp had significantly higher LAB and protease, leu‐aminopeptidase and β‐galactosidase activity (p < .05). The lowest presumptive Vibrio count (VBC) was also observed in intestines of SYN‐fed shrimp. Therefore, we suggested that Lac. plantarum 7–40+ GOS can be used as a synergistic synbiotic for shrimp culture.     

Effects of dietary supplementation with β‐glucan and synbiotics on growth,haemolymph chemistry,and intestinal microbiota and morphology in the Pacific white shrimp

The goal of this study was to investigate the effects of dietary supplementation with β‐glucan and microencapsulated probiotics (Bacillus subtilis or Pediococcus acidilactici) on growth performance, body composition, haemolymph constituents, and intestinal morphology and microbiota of the Pacific white shrimp Litopenaeus vannamei. Four treatment diets [basal diet (C), β‐glucan‐containing diet (β‐glu), β‐glucan plus B. subtilis‐containing diet (β‐glu+Bs), and β‐glucan plus P. acidilactici‐containing diet (β‐glu+Pa)] were fed to L. vannamei for 90 days. Shrimp fed the β‐glu and β‐glu+Pa diets exhibited similar growth performance and body protein content, which were significantly higher than those of shrimp fed the control diet (P < 0.05). No significant differences in haemolymph triglyceride, cholesterol, protein, haemolymph urea nitrogen or chloride were detected among the experimental diets. However, dietary β‐glucan alone increased the haemolymph glucose level and osmolarity (P < 0.05). Synbiotic supplementation had greater effects on intestinal microbiota and morphology than dietary β‐glucan alone. For example, β‐glu+Bs increased the number of intestinal lactic acid bacteria and decreased the number of Vibrio spp. (P < 0.05), and β‐glu+Pa increased the height of intestinal villi.     

Intestinal microbiota of white shrimp,Litopenaeus vannamei,fed diets containing Bacillus subtilis E20‐fermented soybean meal (FSBM) or an antimicrobial peptide derived from B. subtilis E20‐FSBM

The purpose of the present study is to profile the microfloral structure in the intestines of the white shrimp, Litopenaeus vannamei, after being fed a control diet, in comparison with the diets containing Bacillus subtilis E20‐fermented soybean meal (FSBM) or an antimicrobial peptide (AMP) isolated from B. subtilis E20‐FSBM (diets designated FSBMD and AMPD, respectively) for 60 days. Metagenomic data showed detection of eight phyla, 20 classes, 40 orders, 68 families and 96 genera. Despite no statistically significant difference, an evaluation of microbial diversity recorded higher species richness, Shannon–Weaver diversity index and evenness in the AMPD and FSBMD groups, compared to the control diet group. Venn diagrams showed that 58 of the operational taxonomic units (OTUs) were shared among the control, FSBMD and AMPD groups, but additional beneficial microbes were only found in the FSBMD and AMPD groups. In addition to the augmentation of beneficial bacteria in the FSBMD and AMPD groups, the abundance of potential pathogens, Vibrio and Flavobacterium, was lower in the gut of shrimp fed the FSBMD and AMPD. The results clearly suggest that the FSBMD and AMPD have the capability to change the microfloral structure of shrimp intestines and could be used for disease prevention in shrimp aquaculture.     

Dietary nucleotide‐rich yeast supplementation improves growth,innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary nucleotide (NT)‐rich yeast supplementation on growth, innate immunity and intestinal morphology in Pacific white shrimp (Litopenaeus vannamei). Four isonitrogenous and isolipidic practical diets were formulated to contain 0 (control), 10, 30 and 50 g/kg of NT‐rich yeast, respectively. A total of 480 shrimp with an average initial body weight of 1.86 ± 0.02 g were randomly allocated into four groups, with four replicates per group and 30 shrimp each replicate. The results indicated that shrimp fed the diet containing 50 g/kg NT‐rich yeast had significantly higher weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) than those fed the control diet, and the lowest feed conversion ratio (FCR) was observed in the shrimp fed the 50 g/kg NT‐rich yeast supplemental diet. However, there was no significant difference in survival among all treatments. The crude protein of whole shrimp in the 50 g/kg NT‐rich yeast group was higher than that in the control group. Total protein, triglyceride concentrations, the activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly influenced by the dietary NT‐rich yeast supplementation. The activities of serum phenoloxidase (PO) and lysozyme (LZM) of shrimp fed the diet containing 50 g/kg NT‐rich yeast were higher than those in shrimp fed the other diets. Relative expressions of alp and lzm significantly upregulated in the 30 g/kg NT‐rich yeast group compared to the control group. The intestinal fold height and fold width in the 30 g/kg NT‐rich yeast group were significantly higher than those fed the control diet; and the highest microvillus height occurred in the shrimp fed the 50 g/kg NT‐rich yeast diet. In summary, dietary 30–50 g/kg NT‐rich yeast supplementation promotes growth performance, enhances innate immunity and improves intestinal morphology of Litopenaeus vannamei.     

Litopenaeus vannamei fed diets with different replacement levels of fish meal by fish silage: A molecular approach on intestinal microbiota

We investigated the intestinal microbiota in shrimp fed diets with five replacement levels (0, 25%, 50%, 75% and 100%) of fish meal by fish silage. An 8‐week feeding trail was conducted in 15 fibreglass tanks (400 L volume, three replications for each diet) with 50 shrimps (initial weight 0.26 ± 0.03 g) per tank. After 8‐week trial, shrimp fed diets with low replacement levels (0% and 25%) of fish meal by fish silage had significantly enhanced growth performance (p < 0.05). Based on the Illumina sequencing of 15 samples, a total of 5,778 OTUs were obtained and the number of OTUs in each sample ranged from 2,346 to 3,491. OTUs were identified into 23 phyla, and Proteobacteria was the most abundant. At family and genus levels, family Vibrionaceae and genus Photobacterium were more abundant in high replacement level groups (50%, 75% and 100%) than those in low replacement level groups (0% and 25%), which may be related to the worse growth performance of high replacement level groups. However, dietary different levels of fish silage did not make significant difference on intestinal bacterial richness, diversity and metabolism functions (p > 0.05), which indicated the limited impact of dietary fish silage on intestinal microbiota.     

Suitable dietary chitosan improves the growth performance,survival and immune function of tiger shrimp,Penaeus monodon

Different levels of dietary chitosan on growth performance, survival and stress tolerance to air exposure was studied in tiger shrimp, Penaeus monodon. Shrimp (mean initial wet weight about 1.16 g) were fed with six different diets (C0, C0.05, C0.1, C0.2, C0.3 and C0.4) containing six level of chitosan (0%, 0.05%, 0.1%, 0.2%, 0.3% and 0.4% respectively) in triplicate for 60 days. Growth performance [final body wet weight (FBW); weight gain (WG); biomass gain (BG)] of shrimp fed chitosan‐containing diets were higher (P < 0.05) than that of shrimp fed the basal diet, shrimp fed C0.1 diet showed the highest value of growth performance. Survival of shrimp in C0.1 and C0.2 diet groups were higher (P < 0.05) than that of shrimp in C0, C0.05 and C0.4 diet groups but without statistical difference (P > 0.05) in shrimp fed C0.3 diet group. Whole body and muscle lipid contents decreased with increasing dietary chitosan levels. Plasma total cholesterol and triglyceride contents of shrimp fed C0 diet was significantly higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities of shrimp fed C0 diet were higher than those of shrimp fed chitosan‐containing diets. Digestive gland malondialdehyde (MDA) and carbonyl protein contents of shrimp fed chitosan‐containing diets were lower (P < 0.05) than that of shrimp fed C0 diet. Total haemocyte count of shrimp fed C0 diet was lower (P < 0.05) than that of shrimp fed chitosan‐containing diets. On the contrary, the haemolymph clotting time of shrimp fed C0 diet was higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. In conclusion, all results suggested that dietary intake containing 0.1% and 0.2% chitosan enhanced the growth of shrimp, whereas a higher level than 0.3% and 0.4% decreased growth of shrimp. Second‐degree polynomial regression analysis of WG and BG indicated that the optimum supplement of dietary chitosan level should be 0.19–0.21%.     

Opportunistic pathogens are abundant in the gut of cultured giant spiny frog (Paa spinosa)

Pathogen infection poses a serious threat to the survival and commercial quality of cultured Paa spinosa, which provide protection as a substitute for wild P. spinosa. The gut microbiota plays vital roles in host health and immunity. To provide guidance for preventing intestinal diseases of artificial P. spinosa culture, we compared gut microbiota compositions of wild and cultured P. spinosa using high‐throughput sequencing. A total of 11,526 operational taxonomic units (OTUs) were identified from 14,043 sequences from each sample. Cetobacterium, PW3 Bacteroides and some unidentified species from Bacteroidales, Rikenellaceae and Clostridiales were significantly increased in the gut microbiota from cultured P. spinosa, whereas Faecalibacterium and unidentified bacteria from Ruminococcaceae were significantly decreased in the gut microbiota from cultured P. spinosa. According to the gut microbiota composition, we hypothesized that the cultured P. spinosa in Jing'an would exhibit a higher risk of pathogenic infection than those in Cili. These results provided a method to forecast the pathogenic infection risk of cultured P. spinosa, which could guide the artificial culture of and prevent diseases in P. spinosa through gut microbiota.     

Fishmeal levels can be successfully reduced in white shrimp (Litopenaeus vannamei) if supplemented with DL‐Methionine (DL‐Met) or DL‐Methionyl‐DL‐Methionine (Met‐Met)

The main objective of this study was to evaluate the effect of methionine supplementation when reducing fishmeal levels in diets for white shrimp (Litopenaeus vannamei). Tested diets consisted of a positive control with 260 g/kg fishmeal (D1), two negative controls with 100 g/kg fishmeal and no amino acid (AA) supplementation (D2) or supplemented with lysine but not methionine (D3), and four additional diets with 100g/kg fishmeal supplemented with increasing levels of DL‐Met (1.0, 2.0 or 3.0 g/kg) (D4, D5, D6) or Met‐Met (1.0 g/kg) (D7). Each diet was fed to four groups of 30 shrimp for 8 weeks at a daily rate of 70 g/kg body weight. Reduction in fishmeal from 260 g/kg down to 100 g/kg did not significantly affect survival rate, feed conversion ratio (FCR), protein efficiency ratio (PER) or protein retention efficiency (PR%) of white shrimp. However, growth performance (final body weight, FBW; weight gain, WG; specific growth rate, SGR) was reduced when dietary fishmeal level was reduced from 260 g/kg (D1) to 100 g/kg without methionine supplementation (D2). The growth performance (FBW, WG and SGR) of shrimp was significantly increased by supplementation of the 100 g/kg fishmeal diet with increasing levels of DL‐Met (p < .05). Same performance as positive control (D1) was achieved with diets containing 100 g/kg fishmeal and supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. The highest values of growth performance (FBW, WG and SGR) were found in shrimp fed D6 and D7 diets, which were significantly higher than those of shrimp fed D2 and D3 diets (p < .05) but without statistical differences with shrimp fed D1, D4 and D5 diets (p > .05). The highest values of whole‐body and muscle protein contents were found in shrimp fed D1 diet, which were significantly higher than those of shrimp fed all other diets (p < .05). The highest value of intestinal tract proteolytic enzyme activity was found in shrimp fed Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) and 3 g/kg DL‐Met‐supplemented diet (D6) (p < .05). The highest values of apparent digestibility coefficients (ADCs) of dry matter and crude protein were found in Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) (p < .05). Shrimp fed the D1 diet showed the highest value of total essential amino acid (EAA) and was significantly higher than shrimp fed D2–D3 (p < .05) but without significant difference with shrimp fed D4–D7 (p > .05). In conclusion, results showed that same performance can be achieved with diets containing 260 or 100 g/kg fishmeal supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. Moreover, supplementation of limiting methionine in low‐fishmeal diets seems to improve the digestive proteolytic activity, improving digestibility of dry matter and protein, and eventually to promote growth of juvenile white shrimp in fishmeal reduction diets.     

Influence of different dietary carbohydrate sources on the growth and intestinal microbiota of Litopenaeus vannamei at low salinity

Because of the increasing market potential of Pacific white shrimp (Litopenaeus vannamei) and limited availability of coastal areas for production, culture of L. vannamei at low salinity is a growing trend throughout the world. Dietary manipulation could improve the growth performance of L. vannamei at low salinity. This study reared L. vannamei with glucose, sucrose and corn starch as dietary carbohydrate sources, respectively, at a low salinity. The results indicated that the sucrose and glucose feeding groups showed better growth performance. An Illumina‐based sequencing method was used to examine the intestinal bacterial composition and the results indicated that Proteobacteria were the most prevalent members, but abundance of Actinobacteria decreased while Firmicutes increased in the corn starch‐fed group. Furthermore, bacteria related to complex carbohydrate degradation were in lower abundance, whereas the abundance of opportunistic pathogenic bacteria increased in corn starch‐fed group than the other two groups, suggesting that the diet imposes selective pressure on the intestinal microbiota. Complex carbohydrates were not the ideal energy sources for L. vannamei at low salinity because the host has higher energy demand in the stressful conditions while the complex carbohydrate degradation efficiency of the gut microbiota in L. vannamei is limited.     

Effects of dietary astaxanthin on growth,antioxidant capacity and gene expression in Pacific white shrimp Litopenaeus vannamei

Pacific white shrimp Litopenaeus vannamei (1050 individuals with initial weight of 1.01 ± 0.001 g) were fed either control diet or one of six dietary astaxanthin (AX) concentration (25, 50, 75, 100, 125 and 150 mg kg⁻¹) diets for 56 days in 35 tanks (30 shrimp per tank). After 56 days of culture, shrimp‐fed AX125 and AX150 diets had higher (P < 0.05) weight gain, specific growth rate, total antioxidant status and lower (P < 0.05) superoxide dismutase (SOD), catalase (CAT) than shrimp fed control diet. After low dissolved oxygen stress for 1 h, survival rate of shrimp fed AX75, AX100, AX125 and AX150 diets was higher (P < 0.05) than that of shrimp fed control diet. Hypoxia inducible factor‐1α (HIF‐1α), cytosolic manganese superoxide dismutase (cMnSOD) and CAT mRNA expression levels of shrimp fed seven diets were significantly down‐regulated under hypoxia than under normoxia, but their expression levels were higher under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet. About 70‐kDa heat‐shock protein (Hsp70) mRNA expression level of shrimp fed seven diets was significantly up‐regulated under hypoxia than under normoxia, but its expression level was lower under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet.     

Improvement of growth performance,water quality and disease resistance against Vibrio harveyi of postlarval whiteleg shrimp (Litopenaeus vannamei) by administration of mixed microencapsulated Bacillus probiotics

The objective of this study was to investigate the effects of mixed Bacillus on growth, water quality and disease resistance against Vibrio harveyi in whiteleg shrimp (Litopenaeus vannamei). Postlarval shrimp (PL₃₀) were fed with (a) a basal diet (the control), (b) a diet containing mixed freeze‐dried Bacillus probiotics (FB) and (c) addition of mixed microencapsulated Bacillus probiotics (MB) in culture water. Addition of FB and MB probiotics improved (p < .05) growth, feed efficiency, survival and culture water quality (ammonia and nitrite) compared to the control group although there was no difference (p > .05) between the two treated groups. Bacillus numbers in gastrointestinal tracts and culture water of FB‐ and MB‐administrated shrimp were higher (p < .05) than in the control. After a 30‐day culture, shrimp were infected with V. harveyi and monitored for 10 days. A significant reduction (p < .05) in cumulative mortality was observed in FB‐ and MB‐supplemented shrimp (43.24% and 45.05%, respectively), compared to the control (63.06%). This finding demonstrated that administration of microencapsulated probiotics was as effective as freeze‐dried probiotics for improving growth, feed efficiency, survival, Bacillus in gastrointestinal tracts, water quality (ammonia and nitrite) and conferring disease resistance to V. harveyi.     

Debaryomyces hansenii L2‐enriched diet enhances the immunity status,gene expression and intestine functionality in gilthead seabream (Sparus aurata L.)

This study evaluates the effects of the dietary administration of the live yeast Debaryomyces hansenii strain L2 on the immune responses of gilthead seabream for 4 weeks. Cellular immune parameters were measured from serum and head‐kidney leucocytes respectively. The expression levels of immune‐associated genes were quantified by real‐time polymerase chain reaction. In addition, the profile of intestinal microbiota was studied by denaturing gradient gel electrophoresis. The results showed that seabream fed a diet containing D. hansenii had significantly increased cellular immune parameters. The yeast‐supplemented diet up‐regulated the expression of most seabream genes at week 2 and down‐regulated all of them at week 4, except in the head‐kidney. Finally, a reduction in the diversity of the intestinal microbiota was detected in those specimens receiving the yeast‐supplemented diet. These results support the idea that the live yeast D. hansenii strain L2 stimulates the immune system of gilthead seabream.     

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 polysaccharides extracts of algae Ulva rigida on growth,antioxidant, immune response and resistance of shrimp,Litopenaeus vannamei against Photobacterium damselae

This study was carried out to investigate the effect of water‐soluble polysaccharides extract of algae Ulva rigida (WPU) as dietary supplement on growth performance, antioxidant enzyme activity, lysozyme and phenoloxidase activity, and resistance of shrimp (Litopenaeus vannamei) subjected to bacterial infection with Photobacterium damselae. Three replicate groups of shrimp (1.0 g) were fed four diets containing four levels, 0 or control, 0.5, 1 and 1.5 g/kg of WPU for 8 weeks over the growth trial. Thereafter, 30 shrimps from each dietary treatment were infected with bacteria P. damselae to evaluate disease resistance of infected shrimp. The results of this study showed that WPU was effective as a growth promoter for L. vannamei. The best growth rate was observed in shrimp fed 1.5 g/kg of WPU diet. Regarding antioxidant defences, the diets supplemented with three levels of WPU stimulated glutathione peroxidase and catalase activates in experimental shrimps. MDA content of L. vannamei‐fed diet containing WPU 1.5 and WPU 1.0 was lower than WPU 0 and WPU 0. 5 diets. Also, lysozyme and phenoloxidase activities of shrimp receiving WPU at 1.0 and 1.5 level were significantly higher than those fed WPU 0 and WPU 0.5 diets. In addition, using WPU extract in all diets decreased mortality in L. vannamei in a dose‐dependent manner after challenge with P. damselae. These results suggest that incorporation of water‐soluble polysaccharides from green algae U. rigida at 1.5 g/kg doses improves growth and antioxidant activity and enhances the immune responses in shrimp L. vannamei.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

Comparison of L‐lysine·HCl and L‐lysine sulphate in the feed of Penaeus monodon and re‐evaluation of dietary lysine requirement for P. monodon

Two trials were conducted to compare L‐lysine HCl and L‐lysine sulphate regarding its availability to Penaeus monodon, and further evaluate the optimum dietary lysine requirement. In experiment 1, five experimental diets were formulated (D1, D2, D3, D4 and D5), a basal diet (D1), aimed at a low‐lysine concentration (2.22% dry matter), with lysine concentration of the other four diets increasing in two 0.25% L‐lysine intervals from either L‐lysine HCl (D2 and D3) or L‐lysine sulphate (D4 and D5). Each diet was fed at a restricted rate to three groups of 40 shrimp for 74 days. The highest values of growth performance (weight gain, WG; specific growth rate, SGR) and survival were observed with shrimp fed the L‐lysine HCl diet. Feed efficiency (FE) of shrimp fed D2 was significantly higher than that of shrimp fed D1 and D5 (P < 0.05), but without significant difference with shrimp fed D3 and D4 (P > 0.05). In experiment 2, six diets (d1, d2, d3, d4, d5 and d6) were formulated with six graded levels of lysine (2.21%, 2.41%, 2.59%, 2.87%, 3.11% and 3.29% of diet). Each diet was randomly assigned to triplicate groups of 40 shrimp for 74 days. WG, SGR and survival increased increasing levels of lysine up to 2.41% of diet and reached an apparent plateau. Broken‐line model analysis on WG and SGR indicated that the optimum dietary lysine level for optimal growth of shrimp was 2.37% of diet, corresponding to 5.78% of dietary protein. In conclusion, results of this trial suggest that L‐lysine HCl is superior to L‐lysine sulphate when fed to Penaeus monodon and optimal growth can be obtained at lysine levels corresponding to 2.37% of diet, or 5.78% of dietary protein in this specie.     

Growth performance,haematological parameters,antioxidant status and salinity stress tolerance of juvenile Pacific white shrimp (Litopenaeus vannamei) fed different levels of dietary myo‐inositol

A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.     

Histopathological changes in giant freshwater prawn Macrobrachium rosenbergii (de Man 1879) fed with probiotic Bacillus licheniformis upon challenge with Vibrio alginolyticus

The effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the histopathological changes in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g) challenged with known pathogenic strain of Vibrio alginolyticus are reported. Two isocaloric basal diets supplemented with probiotic bacteria B. licheniformis (1.0 × 10⁹ cfu/g feed) and other without probiotic supplementation were fed to the M. rosenbergii juveniles for 45 days. The histological observations revealed no significant changes in the hepatopancreas and gut tissues of both the experimental and the control groups which indicate that the present bacterium is a safe candidate probiont for the host. Prawns were challenged with V. alginolyticus after 45 days of feeding with probiotic diet. The histopathological studies of the hepatopancreas revealed that M. rosenbergii fed with probiotic‐supplemented diet showed less changes as compared to the prawns fed with control diet on second and fourth day of post‐experimental challenge with V. alginolyticus. Histopathological observations revealed that the gills of the prawns fed with control diet were severely affected in comparison to the prawns fed with probiotic‐supplemented diet after challenging with V. alginolyticus. Results from this study revealed the improved protection by dietary incorporation of B. licheniformis in reducing the histopathological manifestations due to V. alginolyticus infection in freshwater prawn.     

Short‐term effects of dietary soybean meal and lactic acid bacteria on the intestinal morphology and microbiota of Atlantic salmon (Salmo salar)

The aim of this study was to evaluate the role of the intestinal microbiota in soybean meal enteritis. Three groups of Atlantic salmon (Salmo salar) were fed for 35 days with different diets: a control diet (CD) containing 510 g kg^?1 fishmeal, diet 1 (D1) containing 378 g kg^?1 of soybean meal and diet 2 (D2) containing D1 supplemented with two viable lactic acid bacteria (LAB). As expected, the fish fed with D1 showed clear signs of distal intestinal inflammation during the study compared with the fish fed CD. For the fish fed with D2, the addition of LAB diminished the inflammation at day 28, but did not abolish it. Microbiological analysis demonstrated that specific bacterial groups were not correlated with the development of enteritis, but were correlated with the three diets. Microbacterium, Pseudomonas, Lactococcus lactis sp. cremoris and Aeromonas VIa were correlated with the CD, Aeromonas VIb and Sporosarcina aquimarina were correlated with D1, and Alcaligenes, Acinetobacter, L. lactis sp. lactis and Carnobacterium maltaromaticum were correlated with D2. Shewanella was not affected by the diet and was present in all fish intestines. Our study suggests that LAB may modulate intestinal inflammation; however, the role of the microbiota in the aetiology of enteritis in fish still requires further study.     

Effects of dietary herbal formulae combined by Astragalus polysaccharides,chlorogenic acid and allicin in different combinations and proportions on growth performance,non‐specific immunity,antioxidant status,vibriosis resistance and damage indexes of Litopenaeus vannamei

In this study, a feeding trial followed by a challenge test was performed to evaluate effects of six herbal formulae which were different combinations of Astragalus polysaccharides (APS), chlorogenic acid (CGA) and allicin on growth performance, non‐specific immune response, antioxidant capacity, disease resistance and biomolecule damage of white shrimp Litopenaeus vannamei. Shrimp were fed seven diets, control diet (basal diet with no herbal formulae) and six herbal formula diets (G1–G6, basal diet supplemented with 0.1% APS + 0.05% allicin, 0.1% APS + 0.1% allicin, 0.1% CGA + 0.05% allicin, 0.1% CGA + 0. 1% allicin, 0.1% APS + 0.1% CGA and 0.1% APS + 0.1% CGA + 0.05% allicin respectively), for 21 days. After that, shrimp were challenged with Vibrio harveyi and then the cumulative mortality of shrimp was recorded for 7 days post challenge. The results showed that there were no significant differences in growth performance among all groups, while the non‐specific immune responses and antioxidant indexes were significantly improved (p < .05) in shrimp fed herbal formula diets when compared to the control. Meanwhile, the lowest cumulative mortality was observed in shrimp fed herbal formula diets with 0.1% APS, 0.1% CGA and 0.05% allicin supplementation after V. harveyi challenge. Additionally, herbal formulae could not cause biomolecule damage to the hepatopancreas of shrimp. In conclusion, these results indicated that synergistic effect of APS, CGA and allicin helped to boost immunity, antioxidant capacity and disease resistance of shrimp without biomolecule damage.