Probiotic efficiency of Bacillus sp. in Labeo rohita challenged by Aeromonas hydrophila: assessment of stress profile,haemato‐biochemical parameters and immune responses

Fish are always susceptible to a wide variety of deadly pathogens which cause a huge loss in aquaculture industries. In this investigation, we have demonstrated the in vivo probiotic efficiency of Bacillus sp. MVF1 (GenBank Acc. No. KP256503) in Labeo rohita challenged with pathogenic strain of Aeromonas hydrophila (MTCC 1739). To check the probiotic potential of the selected bacterial strain, fish were divided into four groups: control, D1, D2 and D3. A total of 100 days (70 days probiotic feeding + 71th day sampling and 28 days challenged test + 29th day sampling) of feeding trial was conducted. To establish the probiotic potential of Bacillus sp. MVF1, certain haematological parameters (haemoglobin, total erythrocyte and leucocyte count), serum biochemical parameters (total protein, albumin and globulin), immune parameters (serum lysozyme and total IgM levels) and hepatic stress profile (malondialdehyde production, superoxide dismutase and catalase activity) have been measured. Our results demonstrated that red blood cell number, white blood cell number and haemoglobin content were much higher in D2 group fish compared to other groups and control fish. Similarly, total protein contents, albumin concentration, globulin concentration, lysozyme activity and IgM production were also recorded to be highest in D2 group fish. This finding clearly indicated the probiotic potential of Bacillus sp. MVF1 in L. rohita. Furthermore, our results also demonstrated that 1 × 10⁷ CFU g^?1 feed (D2) provides better immunity compared to 1 × 10⁵ (D1) and 1 × 10⁹ (D3). Due to beneficial effects, the bacterium Bacillus sp. MVF1 might be useful in aquaculture industries to reduce the disease susceptibility.     

Effects of Lactobacillus pentosus on the growth performance,digestive enzyme and disease resistance of white shrimp,Litopenaeus vannamei (Boone, 1931)

The objective of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) strains isolated from digestive tract of white shrimp Litopenaeus vannamei. Eighteen LAB colonies were isolated and one bacterium was found capable of producing three extracellular enzymes (protease, cellulose and lipase) simultaneously and exhibited antagonistic activity against shrimp pathogens (Vibrio vulnificus, V. rotiferianus and V. campbellii). The putative probiotic strain AS13 was identified as Lactobacillus pentosus based on 16S rRNA sequencing. The L. vannamei were fed diet containing 0 (control), 10⁶, 10⁷ and 10⁸ CFU g^?1 bacterial cells of AS13 for 28 days. The results showed that supplementation of L. pentosus significantly improved the growth performance and feed utilization in the treated groups over the control. Similarly, digestive enzyme activities were elevated in the intestines of treated groups. Moreover, feeding of supplemented diets containing AS13 significantly reduced the mortality rate caused by pathogenic Vibrio species (V. vulnificus, V. rotiferianus and V. campbellii). Our results indicated L. pentosus AS13 addition at 10⁷ CFU g^?1 can effectively enhance the growth performance, feed utilization, digestive enzymes and disease resistance of L. vannamei in the laboratory condition.     

Effects of diets supplemented with either individual or combined probiotics,Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, on the immune response and disease resistance of the mud crab,Scylla paramamosain (Estampador)

Two trials, including firstly, diets incorporating individual or combined species of probiotics, Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, were fed to the mud crab, Scylla paramamosian, for 28 days to evaluate the growth, immunity and disease resistance, and secondly, the optimal level of B. subtilis E20 in the diet by tracking the growth, immunity and disease resistance after 28 days of feeding were analysed. No significant differences in growth, total haemocyte counts, respiratory bursts, superoxide dismutase, or glutathione peroxidase were found in the two trials. Crabs fed the B. subtilis E20‐containing diet (D20) had significantly increased phenoloxidase (PO) activity, phagocytic activity (PA) and disease resistance compared with crabs fed the control and the mixed probiotics diet (MD). The mortality of crabs fed the D20 was significantly lower than that of crabs fed the L. plantarum 7‐40‐containing diet. L. plantarum 7‐40 had a great inhibitory effect on the growth of B. subtilis E20, which may have led to the decreased probiotic effect of the MD. An analysis of the optimal level of B. subtilis E20 in the diet showed that crabs fed the B. subtilis E20‐containing diet at a level of 10^9–10 cfu kg^?1 had better immune response of PO and PA, and disease resistance in the second trial.     

Optimal dietary alpha‐linolenic acid/linoleic acid ratio improved digestive and absorptive capacities and target of rapamycin gene expression of juvenile grass carp (Ctenopharyngodon idellus)

Growth performance, digestive and absorptive capacities and target of rapamycin (TOR), ribosomal protein S6 kinase 1 (S6K1) and eIF4E‐binding protein (4E‐BP) gene expression in the hepatopancreas and intestine of juvenile grass carp (Ctenopharyngodon idellus) fed graded ratios of dietary alpha‐linolenic acid/linoleic acid (ALA/LNA) (0.01, 0.34, 0.68, 1.03, 1.41, 1.76 and 2.15) for 60 days were investigated. The results showed that ALA/LNA ratio of 1.03 significantly improved (i) per cent weight gain (PWG) and feed efficiency, (ii) hepatopancreatic trypsin, chymotrypsin, lipase, amylase and intestinal creatine kinase (CK) activities, (iii) hepatopancreatic trypsinogen‐2 and chymotrypsinogen mRNA levels. Meanwhile, fish fed with ALA/LNA ratio of 0.68 significantly enhanced, (iv) Na⁺/K⁺‐ATPase and γ‐glutamyl transpeptidase activities in whole intestine, and alkaline phosphatase activities in the proximal intestine (PI) and distal intestine, (v) amylase, intestinal Na⁺/K⁺‐ATPase alpha‐subunit isoform 1, Na⁺/K⁺‐ATPase alpha‐subunit isoform 8 and CK mRNA abundances, (vi) TOR and S6K1 gene expression in the hepatopancreas and intestine of juvenile grass carp. Based on the quadratic regression analysis of PWG, cholecystokinin and leptin contents in the PI, optimal dietary ALA/LNA ratio of juvenile grass carp (8.78–72.00 g) was estimated to be 1.08, 1.19 and 1.05, respectively.     

Effect of cortisol and triiodothyronine bath treatments on the digestive enzyme profile and growth of Catla catla larvae during ontogenic development

Digestive enzyme profile is a good indicator of the nutritional and health status of the fish. The present investigation aims to evaluate the effect of exogenous bath treatment of hormones, cortisol and triiodothyronine, on the digestive enzyme activities and growth of carp Catla catla (Ham.) during ontogenic development. Catla larvae (4 days old) were given bath treatment with cortisol (hydrocortisone, 0.2 mg L^?1), 3,5,3′‐triiodothyronine (T3, 2.5 mg L^?1) and a combination of cortisol and T3 for 30 min. Digestive enzyme profile was recorded on every third day and was continued for 30 days. Larvae were fed with live food for initial 14 days and then weaned to mix feeding of live food and prepared diet. Significantly (P < 0.05) higher amylase, total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were found in the hormone‐treated groups compared to the control one during ontogenic development. Among the treated groups, amylase activity was highest in cortisol‐treated larvae. Total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were significantly (P < 0.05) higher in larvae exposed at combined treatment of cortisol and T3 compared to the other two groups in most sampling days. Average length, weight and specific growth rate of treated larvae were higher compared to the control one. The combined bath treatment of cortisol and triiodothyronine influenced the digestive enzyme activities of catla larvae and thereby enhanced the growth at early developmental stage. This helps the larvae to overcome the problems associated with early developmental stage.     

Effect of dietary administration of Psychrobacter sp. on the growth,feed utilization,digestive enzymes and immune responses of grouper Epinephelus coioides

A 60‐day feeding trial was conducted to assess the effect of incorporating dietary Psychrobacter sp., one predominant bacterium with antagonistic activity in the gut of fast‐growing grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune responses of E. coioides at regular monthly intervals. The fish were fed with control diet (without probiont) or probiotic diet supplemented with 1.0 × 10⁸ CFU g^?1Psychrobacter sp. No significant improvement in weight gain (WG) and specific growth ratio (SGR) were observed in the probiotic group, but a significant improvement in feed conversion ratio (FCR) was observed in fish fed the probiotic diet for 60 days. Although not statistically significant, enhanced hepatopancreatic protease and lipase activities and intestinal amylase activity were observed in probiotic group at day 60. A slightly increase in phagocytic activity and phagocytic index was observed in fish fed probiotic diet for 60 days. Serum lysozyme activity showed no significant differences between the probiotic group and the control group during the whole experiment period, while an increasing trend of superoxide dismutase (SOD) activity in the probiotic group was observed. Serum complement component 3 (C3) level of the probiotic group was higher than that of control in the whole feeding period, while complement component 4 (C4) level of the probiotic group was significantly higher than that of the control at day 60. The results of the current study firstly demonstrated potential for Psychrobacter sp. as a novel probiont to improve feed utilization, digestive enzymes and innate immunity in E. coioides.     

Protective effects of basil (Ocimum basilicum) ethanolic extract supplementation diets against experimental Aeromonas hydrophila infection in common carp (Cyprinus carpio)

This study evaluated the effects of basil (Ocimum basilicum) leaves extract supplement on growth, blood parameters and resistance to Aeromonas hydrophila in Cyprinus carpio fingerlings. Basil leaves were mixed thoroughly with feed at 0, 100, 200, 400, 800 and 1600 mg kg^?1 of diet and were fed in triplicate group for 2 months. At the end of this period, growth performance and survival were determined. Then fish were challenged intraperitoneally with A. hydrophila and mortalities were recorded up to 10 days post challenge. Results indicated that WBC, RBC, Ht, Hb, total protein, albumin and globulin were higher (P < 0.05) in fish fed diets containing O. basilicum compared with the control. Highest specific growth rate values were observed in 400 mg kg^?1 group. Feed conversion ratio was significantly lower (P < 0.05) in fingerlings fed 400 and 800 mg O. basilicum kg^?1 dry diet. After 10 days post challenge, total protein, WBC, RBC and Hb were significantly higher in 400 mg kg^?1 compared with other groups. These results reveal that a dietary O. basilicum leaves extract of 400 mg kg^?1 fed for 60 days leads to increased growth performance and survival rate as well as improved feeding efficiency in common carp fingerlings rendering them more resistant against infection by A. hydrophila.     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.     

Effects of Pediococcus pentosaceus supplementation on growth performance,intestinal microflora and disease resistance of white shrimp,Litopenaeus vannamei

Litopenaeus vannamei is economically important shrimp species in worldwide aquaculture. This study was conducted to assess the effect of different levels of probiotic Pediococcus pentosaceus (PP) on growth performance, feed utilization, digestive enzyme activity, intestinal microflora count and body composition of L. vannamei. Four diets containing different concentrations [0 (PP0), 10⁶ (PPI), 10⁷ (PPII) and 10⁸ (PPIII) CFU/g] of PP were formulated. After 8 weeks feeding trial, the experimental shrimps were challenged with Vibrio anguillarum and noted the surveillance. At the end of the feeding trial, the obtained results revealed a significant increase (p < .05) in final body weight, final length, weight gain (WG), survival rate, protease and amylase activities, lactobacillus sp. and Bacillus sp. intestinal count, total haemocyte counts (THC) and lysozyme activity in PPIII group when compared with the other groups. Similarly, WG, amylase activity, Bacillus sp. count, THC and lysozyme activity were significantly enhanced in case of PPII compared to the control group (p < .05). Interestingly, FCR and Vibrio sp. counts were significantly decreased in case of PPIII group when compared to the other groups (p < .05). Also, significant level of surveillance was noted in the challenging test with V. anguillarum. These results suggested that the P. pentosaceus improved the growth performance, digestive enzyme activity, immunity and tolerance against V. anguillarum of L. vannamei.     

Cross‐effects of dietary probiotic supplementation and rearing temperature on growth performance,digestive enzyme activities,cumulative mortality and innate immune response in seabass (Dicentrarchus labrax)

Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross‐effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 10⁸ CFU/kg diet), and tested against a non‐supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α‐amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.     

Effects of dietary phenylalanine on growth,digestive and brush border enzyme activities and antioxidant capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idella)

This study investigated the effects of phenylalanine on growth, digestive and absorptive ability and antioxidant status of young grass carp (Ctenopharyngodon idella). Young grass carp were fed diets containing 3.4 (basal diet), 6.1, 9.1, 11.5, 14.0 and 16.8 g phenylalanine kg^?1 diet with a fixed of 10.7 g tyrosine kg^?1 diet for 8 weeks. Percent weight gain (PWG), feed efficiency and feed intake of fish were the lowest in fish fed the basal diet (P < 0.05). Trypsin, lipase and amylase activities in the hepatopancreas, and antioxidants including glutathione contents and glutathione reducase activities in the hepatopancreas and intestine were all the highest in fish fed 11.5 g phenylalanine kg^?1 diet (P < 0.05). Trypsin, chymotrypsin and amylase activities in whole intestine, and creatine kinase, Na⁺, K⁺‐ATPase and alkaline phosphatase activities in the proximal intestine, and superoxide dismutase activities in the hepatopancreas and intestine were all the highest when phenylalanine at level of 9.1 g kg^?1 diet (P < 0.05). In conclusion, phenylalanine improved growth, digestive and absorptive ability, and antioxidant capacity of young grass carp. The phenylalanine requirement of young grass carp (256–629 g) based on PWG was 10.4 g kg^?1 diet or 3.44 g 100 g^?1 protein.     

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.     

Probiotic activity of bacteria associated with egg capsules of Concholepas concholepas (common name ‘Loco’)

Egg capsules of Concholepas concholepas were analysed microbiologically, showing the presence of bacteria inside the capsules. The bacteria were isolated and identified, and their probiotic potential on early larvae of C. concholepas was evaluated. From 32 capsules that were examined, 45 bacterial strains were isolated, of which 67% belonged to the genus Bacillus. Survival bioassays were carried out with all the isolated bacterial strains on C. concholepas larvae. It was found that 71% of the bacteria had positive effects on larval survival. This research determined that bacteria of the genera Bacillus and Vibrio present in the capsules have probiotic potential. The bacteria that led to the greatest larval survival were Bacillus pumilus, which may be used in the cultivation of C. concholepas, because one of the limiting factors for the cultivation of this species is larval survival; thus, we propose that the probiotic strains identified in this study will be used during the cultivation of C. concholepas to enhance the chances of larval survival.     

Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia

We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 10⁴ cfu mL^?1 (Treatment 1) and 1.0 ×10⁵ cfu mL^?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 10⁷ cfu g^?1 (Treatment 1) and 1.0 × 10⁸ cfu g^?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.     

Dietary Lactobacillus plantarum supplementation decreases tissue lead accumulation and alleviates lead toxicity in Nile tilapia (Oreochromis niloticus)

Pollution by the heavy metal lead (Pb) has become a threat to both aquaculture and food safety. In this study, the protective effects of a selected probiotic with good Pb binding capacity (Lactobacillus plantarumCCFM8661) against waterborne Pb exposure were evaluated in Nile tilapia (Oreochromis niloticus). One hundred and eighty fish were divided into four groups as control, Pb‐only, Pb‐plus‐CCFM8661 and CCFM8661‐only. Fish were exposed to a dose of waterborne Pb level at 1 mg L^?1 for 4 weeks, and the probiotic was administered at 10⁸ CFU g^?1 in fish diet twice daily. The results showed that dietary supplementation of CCFM8661 ameliorated the growth performance and prevented the death of Pb‐exposed fish. Cotreatment with Pb and CCFM8661 also effectively decreased Pb accumulation in kidney, liver, gonad, brain, gills and muscle. Moreover, the administration of this probiotic alleviated Pb‐induced oxidative stress, recovered digestive enzyme activities and blood δ‐aminolevulinic acid dehydratase activity, reversed alterations in innate immune status and decreased the frequencies of the nuclear abnormalities in peripheral blood erythrocytes of fish. These results suggested that CCFM8661 may be a novel dietary supplement against Pb toxicity at least in tilapia.     

Evaluation of Lactobacillus plantarum as a water additive on host associated microflora,growth, feed efficiency and immune response of giant freshwater prawn,Macrobrachium rosenbergii (de Man, 1879)

A 90‐day experiment was conducted by rearing 1020 prawn juveniles (0.54 ± 0.03 g) in water supplemented with three different concentrations of probiotic bacteria viz. T1 (10⁷ cfu L^?1), T2 (10⁸ cfu L^?1), T3 (10⁹ cfu L^?1) and the control (C) (unsupplemented water), to evaluate probiotic effect of Lactobacillus plantarum. In the present study, the growth parameters (WG%, SGR) and feed utilization parameters (FCR, PER) significantly improved (P < 0.05) in T3. The growth and feed utilization parameters though improved marginally in T1 and T2, the difference was not significant (P > 0.05) compared to the control. The gastro‐intestinal Lactobacillus sp. count increased significantly (P < 0.05) in all the treatment groups, whereas the decrease in harmful bacteria was significant (P < 0.05) in T3 compared to the control. Similarly, the Lactobacillus sp. count in culture water increased significantly (P < 0.05) in all the experimental groups, whereas the decrease in harmful bacteria was significant (P < 0.05) in T2 and T3. The immune parameters (THC, PO and RB activity) and clearance efficiency significantly improved (P < 0.05) in T3 with concurrent decrease (P < 0.05) in cumulative mortality against Aeromonas hydrophila challenge. However, water quality did not improved (P > 0.05) in any of the treatment groups. The results indicate that Lactobacillus plantarum at a minimum concentration of 10⁹ cfu L^?1 could be used as water additive to confer its probiotic effect in prawn, Macrobrachium rosenbergii. Moreover, future studies with higher probiotic concentrations should be conducted for its efficient commercial scale field application.     

Pathogenic threats and probiotic use in larviculture of the scallop,Pecten maximus

During a series of experiments, bacteriological elements in scallop larval rearing were investigated: larvae susceptibility to pathogens as a function of their age, and the use of probiotic bacteria during larviculture. Younger larvae (d5 PF) were highly more susceptible to pathogenic‐challenge than their older siblings, which were challenged at an older age (d15 PF). A challenge with 10⁴ CFU mL^?1 of V. pectenicida killed 100% of d5 PF larvae 7 days following challenge, yet killed only 9% of d15 PF larvae 9 days following challenge. Use of the probiotics Phaeobacter gallaeciensis, Alteromonas macleodii 0444 and Neptunomonas sp. 0536, provided for larger larvae, a high yield of competent larvae and, perhaps more importantly, protection against pathogen‐challenge similar to levels achieved from antibiotic use. When challenged with V. pectenicida, d29 survivals were 20.3%, 85.1% and 75.0% respectively for control (no probiotic), antibiotic treated, and ‘probiotic mix’ administered larvae. Use of potential probiotic Pseudoalteromonas sp. D41 appeared to hinder scallop larvae. Future use of probiotics in scallop larval rearing would benefit from combined use of P. gallaeciensis, A. macleodii 0444 and Neptunomonas sp. 0536.     

Effects of supplementation of decapod zoea to Artemia basal diet on fatty acid composition and digestive gland histology in common octopus (Octopus vulgaris) paralarvae

The present study aimed to evaluate the effect of the supplementation of different crab zoeas to enriched Artemia basal diet for Octopus vulgaris paralarvae during the first month of life. Paralarvae were fed using enriched Artemia nauplii alone and Artemia co‐fed either first zoea stages of Grapsus adscensionis or Plagusia depressa. The experiment was carried out over a period of 28 days, in 0.12 m³ tanks with a flow‐through rearing system. Growth in dry weight as well as mantle length and width were assessed weekly. Additionally, prey and paralarvae fatty acid composition and digestive gland (DG) histology were evaluated. Addition of low amounts of crab zoeas (approx. 100 indv. L^?1 day^?1) provided during critical life stages of O. vulgaris proved to be good enough to improve paralarvae growth and survival in comparison with those fed exclusively on enriched Artemia. These results were supported by the finding of a higher number of glycoprotein absorption vacuoles in the DG from paralarvae co‐fed crab zoeas, suggesting a higher feeding activity. In addition, fatty acid analysis of crab zoea showed that these are good sources of dietary arachidonic and eicosapentaenoic acids during the octopus planktonic life stage, whereas the low docosahexaenoic (DHA) content suggests the use of additional DHA sources or higher zoea densities to meet paralarvae nutritional demand to carry out a successful metamorphosis to benthic life.     

Larval feeding behaviour of angel fish Pterophyllum scalare (Cichlidae) fed copepod Eucyclops serrulatus and cladoceran Ceriodaphnia quadrangula

Three different live diets, Eucyclops serrulatus, Ceriodaphnia quadrangula and equal combination of E. serrulatus copepodid and C. quadrangula, were offered to angelfish (Pterophyllum scalare) larvae viz 1‐week, 2‐week and 3‐week old at prey densities of 2, 5 and 10 individuals mL^?1. Results showed that 1‐week‐old P. scalare larvae consumed E. serrulatus copepodid at a rate of 31.3–56.7 ind. h^?1, C. quadrangula at 8.0–12.0 ind. h^?1, and mixture of E. serrulatus and C. quadrangula at 20.7–40.7 ind. h^?1. For 2‐week‐ and 3‐week‐old larvae, consumption rate increased accordingly. The electivity indices (E) of P. scalare (1‐week‐old larvae) for E. serrulatus copepodid were +0.18, +0.23 and +0.22 at prey densities of 2, 5 and 10 ind. mL^?1 respectively. Tendency towards E. serrulatus copepodid consumption reduced by aging P. scalare as indicated by the E values for 2‐ and 3‐week‐old larvae. However, growth and survival of P. scalare larvae was greatest when fed on combination of copepod E. serrulatus and C. quadrangula.     

Deficiency of dietary niacin decreases digestion and absorption capacities via declining the digestive and brush border enzyme activities and downregulating those enzyme gene transcription related to TOR pathway of the hepatopancreas and intestine in young grass carp (Ctenopharyngodon idella)

To investigate the effects of niacin on growth, digestion and absorption capacity, and the potential mechanism for digestive and brush border enzyme activities, grass carp (Ctenopharyngodon idella) (256 ± 0.41 g) were fed diets containing 3.95 (basal diet group), 14.92, 24.98, 35.03, 44.97 and 55.01 mg niacin kg^?1 diet for 8 weeks. Results indicated that percentage weight gain (PWG), feed intake and feed efficiency were the lowest in basal group (P < 0.05). Similarly, niacin deficiency decreased hepatopancreas trypsin, chymotrypsin, lipase and amylase activities (P < 0.05), intestinal Na⁺, K⁺‐ATPase, alkaline phosphatase, γ‐glutamyl transpeptidase and creatine kinase (CK) activities, the cholecystokinin (CCK) content in proximal intestine (PI) and growth hormone content in serum (P < 0.05). Furthermore, niacin deficiency downregulated gene expression of hepatopancreas trypsinogen 1, trypsinogen 2, chymotrypsinogen and amylase, intestinal Na⁺, K⁺‐ATPase alpha subunit isoform 1, Na⁺, K⁺‐ATPase alpha subunit isoform 8 and CK, and target of rapamycin (TOR) and S6 kinase 1 (S6K1) of hepatopancreas and intestine (P < 0.05), whereas upregulated eIF4E‐binding protein (4EBP) gene expression (P < 0.05). The niacin requirement for young grass carp (256–689 g) based on PWG, hepatopancreas trypsin activity and Na⁺, K⁺‐ATPase in PI was 34.01, 35.10 and 42.08 mg kg^?1 diet, respectively.