Effects of dietary 60 g kg−1 dried distiller's grains in least‐cost practical diets on production and gut allochthonous bacterial composition of cage‐cultured fish: comparison among fish species with different natural food habits

Dried distiller's grain (DDG) is considered as an alternative ingredient of dietary feed due to its high contents of protein, fibre and fat. In this study, 60 g kg^‐1 of DDG was used to feed grass carp (Ctenopharyngodon idella), bluntnose black bream (Megalobrama amblycephala), gibel carp (Carassius gibelio) and black carp (Mylopharyngodon piceus) for 8 weeks, and its effect on fish production and gut allochthonous microbiota was investigated for the development of a suitable fish feed high in nutrients and low in cost for polyculture freshwater fish. DDG supplementation resulted in the less weight gain and higher feed conversion ratio of black carp (P < 0.05), but had no significant effects on other fish or parameters. PCR–denaturing gradient gel electrophoresis (DGGE) analysis indicated that all four fish species had some common and unique bacteria in their digestive tracts, and the gut microbiota of bluntnose black and gibel carp fed the control diet and DDG diets were very similar (Cs > 91%); of them, the total counts of intestinal bacteria studied by qPCR increased in grass carp (P < 0.05) and depressed in black carp (P < 0.05) when fed dietary DDG. Thus, we assumed that dietary DDG modulated production and gut microbiota of fish in a host‐specific way.     

Effects of dietary fishmeal replacement with Spirulina platensis on the growth,feed utilization,digestion and physiological parameters in juvenile gibel carp (Carassis auratus gibelio var. CAS III)

A 60‐day growth experiment was conducted to investigate the effect of dietary replacement of fishmeal by Spirulina platensis on growth performance, digestion and physiological parameters in juvenile gibel carp (5.0 ± 1.0 g). Four diets were formulated to replace 0 (SP0), 50% (SP50), 75% (SP75) and 100% (SP100) of dietary fishmeal protein by S. platensis respectively. Other two diets based on SP100 were supplemented with 2% dicalcium phosphate (SP100 + P) or 0.28% coated lysine (SP100 + Lys). The results showed that no significant differences of specific growth rate (SGR), feed efficiency (FE), feeding rate (FR) and protein efficiency ratio (PER) were observed between SP0 group and the replacement groups (SP50, SP75 and SP100). FE and PER of fish‐fed SP100 + Lys diet were significantly higher than the fish‐fed SP100 diet (p < .05). Apparent digestibility coefficients (ADCs) of dry matter, protein, energy and phosphorus increased significantly with the increasing S. platensis inclusion. Meanwhile, fish from SP100 + P or SP100 + Lys group had higher ADCs of nutrients compared to the SP100 group (p < .05). Compared with SP0 diet, fish‐fed S. platensis diets had higher activities of plasma superoxide dismutase, hepatopancreas total antioxidant capacity, plasma alkaline phosphatase and plasma lysozyme, as well as the lower content of plasma malondialdehyde. The present results indicated that dietary fishmeal could be fully replaced by S. platensis without any negative effects on growth performance of gibel carp. Supplementation of lysine in the S. platensis replacement diet could further improve the feed utilization of gibel carp.     

Effects of natural dietary carotenoids on skin coloration of false Clownfish (Amphiprion ocellaris Cuvier, 1830)

This study was performed to evaluate the effect of dietary natural carotenoid sources on skin colour enhancement of false clownfish Amphiprion ocellaris. The juvenile fish (initial body wt. 0.30 g) were fed with four experimental diets including (a) commercial feed (reference diet), (b) moist feed, (c) sweet potato (potato diet) and (d) dried gut weed Enteromorpha sp. (gut weed diet). Sweet potato and gut weed were used to boost up β‐carotene levels in the diets. There was no significant difference in final weight (0.51 ± 0.02 – 0.61 ± 0.01 g) and length (2.80 ± 0.02 cm) of fishes among treatments (p > 0.05) after 8 weeks. The survival rate of the fish in all dietary treatment was greater than 89%. Principal component analysis results showed that fish fed potato and gut weed diets performed brighter colour in skin with more orangeness, body and accumulated β‐carotene levels were higher than those fish fed with reference diet. Indicator a* value for the redness of fish fed potato diet (16.18 ± 0.59) and gut weed (14.36 ± 2.14) was also higher than fish fed reference diet (10.92 ± 0.82). The result of this study provided key information for developing dietary colour enhancement of ornamental fish by using cost‐effective feed ingredients (potato and gut weed) as natural supplemental carotenoid sources.     

Formulated diet alters gut microbiota compositions in marine fish Nibea coibor and Nibea diacanthus

Nibea, an economical marine fish, is generally fed on trash fish (the low‐value fish), which can cause high feed costs and waste pollutions in high‐density aquaculture. To assess the effect of formulated diet on the gut microbiota in Nibea coibor and Nibea diacanthus, denaturing gradient gel electrophoresis, clone libraries analysis and Illumina sequencing of the 16S ribosomal RNA gene were used in this study. Two Nibea fishes were both dominated by Gammaproteobacteria (especially Photobacterium) and shared a set of gut microbiota, including Bacilli, Mollicutes, Alphaproteobacteria and Fusobacteriia. Statistical analyses revealed that formulated diet led to lower feed conversion ratio (p < 0.001), lower abundance of Vibrio (p = 0.040) and infectious diseases pathways (p = 0.001), higher abundance of polysaccharide‐degrading bacterium Cellvibrio (p = 0.006) in two Nibea species, with higher weight gain rate (p = 0.023) and microbial diversity (Shannon, p = 0.049 and Simpson, p = 0.044) and more carbohydrate metabolism (p = 0.020) observed in N. coibor. The distribution and correlation network of 17 potential short‐chain fatty acid producing bacteria were obtained and visualized in all treatment groups. The results reveal that formulated diet has beneficial effects on the gut microbial ecology in two Nibea fishes, which suggests the possibility of replacing trash fish diet with formulated diet in Nibea aquaculture.     

The effect of dietary chitin on the autochthonous gut bacteria of Atlantic cod ( Gadus morhua L.)

In the present study the impact on autochthonous (adherent) bacteria in proximal intestine (PI) and distal intestine (DI) of Atlantic cod ( Gadus morhua L.) was evaluated following feeding of a control diet and a diet supplemented with 5% chitin. The autochthonous gut bacteria were investigated using denaturing gradient gel electrophoresis (DGGE). Analysis of the microbiota associated with PI and DI of Atlantic cod indicate that dietary chitin modulate the intestinal bacterial community. For example, band 25 ( Escherichia coli–like), band 14 ( Anaerorhabdus furcosa–like) and band 29 (uncultured bacterium–like) in PI were depressed by dietary chitin (P < 0.05). The number of bands (23.7 ± 5.4) in DI of fish fed chitin was marginally higher than the control fish (16.7 ± 2.1) (P = 0.065), and the relative abundances of band 6 (swine faecal bacterium–like) were marginally stimulated by dietary chitin (P = 0.095). Furthermore, the present study reports several novel sequences not previously reported in the gastrointestinal tract of Atlantic cod. Whether the dietary effect of chitin on gut bacterial community has any positive effect of fish health merits further investigations.     

Impacts of diet on hindgut microbiota and short‐chain fatty acids in grass carp (Ctenopharyngodon idellus)

Diet is known to influence intestinal microbiota in fish, but the specifics of these impacts are still poorly understood. Different protein/fibre ratio diets may result in differing structures and activities of gut microbiota. We examined the hindgut microbiome of grass carp (Ctenopharyngodon idellus) fed three different diets: fish meal (FM, high protein – low fibre), Sudan grass (SG, high fibre – low protein) and compound feed (CF, intermediate). Microbial profiles of fish fed on FM were significantly different from profiles of fish fed CF and SG (F = 18.85, p < .01). Cetobacterium, known to be positively associated with protein digestion, was the dominant microbial group in FM samples (approximately 75.7%), while Lachnospiraceae and Erysipelotrichaceae, thought to be involved in fermentation of plant polysaccharides, were dominant in CF and SG samples (46.8% and 42.9% respectively). Network analyses indicated that the abundance of Lachnospiraceae and Erysipelotrichaceae was in a significantly positive correlation (r = .895, p = .001). Short‐chain fatty acid (SCFA) levels may indicate that the digestibility of diet by microbiota in the grass carp gut decreased from FM to SG (FM>CF>SG). Overall low SCFA levels indicate that hindgut fermentation probably provides a low proportion of energy requirements in grass carp.     

Effects of dietary supplementation of probiotics on the growth,activities of digestive and non‐specific immune enzymes in hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀)

Effects of Bacillus cereus BC‐01, Lactobacillus acidophilus LAG01, Clostridium butyricum CBG01 and their combinations as supplementation on the growth performance, digestive enzyme activities and serum non‐specific immunity of hybrid grouper (Epinephelus lanceolatus ♂×E. fuscoguttatus ♀) were assessed. Seven different diets, that is one control diet (basal feed without any probiotics, CT) and six treatment diets containing single B. cereus (Bs), L. acidophilus (Ls) and C. butyricum (Cs) at 1.0 × 10⁹ cfu/kg feed, and also their combinations in equal proportion at 1.0, 2.0 and 3.0 × 10⁹ cfu/kg feed (BLC1, BLC2 and BLC3) were prepared respectively. After 60‐day feeding trial, the final weight, specific growth rate,food consumption, food conversion efficiency and apparent digestibility coefficient of fish in Ls and BLC3 were significantly higher compared with the control (p < .05). The activities of pepsin and trypsin in the intestine of fish for Ls and BLC3 were significantly higher relative to the control (p < .05). Relative to controls, significantly enhanced amylase and lipase activities in proximal intestine except for Cs and BLC1 and lipase activities in distal intestine except for Cs were observed (p < .05). Meanwhile, activities of superoxide dismutase in the serum of fish for all treatments, lysozyme and catalase in Ls and BLC3, and glutathione peroxidase except for Cs were significantly enhanced (p < .05). Based on the above, dietary supplementation of single L. acidophilus at 1.0 × 10⁹ cfu/kg or combination of three strains at 3.0 × 10⁹ cfu/kg for hybrid grouper is recommended.     

Evaluation of live duckweed,Wolffia globosa as an allochthonous feed for Labeo rohita fry during nursery rearing

A 40 days feeding trial with Labeo rohita (rohu) fry was conducted in six outdoor cemented tanks (5m × 4m × 1 m; 6–8 cm soil base) to establish the relative impact of on‐farm grown live Wolffia globosa (a duckweed with crude protein of 28.57%) and a formulated iso‐nitrogenous feed (crude protein 28.37%) on growth performance, survival, health and overall quality of fingerlings. The experimental tanks were dried, limed at 250 kg/ha, filled with ground water and subsequently applied with cow dung (soaked overnight) and mustard oil cake at 1,000 kg/ha and 50 kg/ha respectively. The fry were stocked at 0.3 millions/ha in six prepared tanks, and fed live Wolffia (T1) and formulated feed (T2). Results of the study indicate better performance (p < 0.05) of fry in terms of final mean weight (7.83 g), length (8.82 cm), daily growth index (6.51 g %/day) and survival (89.5%) than those fed on formulated feed. Furthermore, significantly improved (p < 0.05) feed utilization parameters viz. apparent feed conversion ratio (AFCR), apparent protein conversion efficiency (APCE), apparent protein efficiency ratio (APER); haematological parameters viz. total erythrocyte count (TEC), total leucocyte count (TLC), packed cell volume (PCV), plasma glucose and protein of fry were also observed in T1 (live Wolffia). Overall findings from this study hints towards the possible utilization of on‐farm grown live Wolffia as a suitable nutrient source for fry rearing of L. rohita.     

Evaluation of growth,feed utilization efficiency and immune parameters in tiger shrimp (Penaeus monodon) fed diets supplemented with or diet fermented with gut bacterium Bacillus sp. DDKRC1. isolated from gut of Asian seabass (Lates calcarifer)

An experiment was conducted to assess the effect of supplementation of cellulolytic gut bacterium with plant protein‐based diet on growth, digestibility and digestive enzyme status of tiger shrimp, Penaeus monodon. Bacillus sp. DDKRC1. (JN641289), a potential cellulolytic bacterium, isolated from the gut of Asian seabass was used as feed supplement in this experiment. The shrimps of group I, II and III were fed with control diet (without bacterial supplement, D1), diet supplemented with live Bacillus sp. DDKRC1. at 2.94 × 10⁷ cfu per 100 g feed (D2) and diet fermented (48 hr) with same bacteria (D3) respectively. At the end of the 42 days experiment, shrimps of group II showed significantly better (p < .01) growth and protein efficiency ratio (PER) and lower (p < .01) feed conversion ratio (FCR) as compared to group I and III. Dry matter, cellulose, hemicellulose and lipid digestibilities were significantly (p < .01) higher in group II than other two groups. Cellulase, amylase and protease activities in the gastrointestinal (GI) tract were significantly (p < .01) higher in group II as compared to other two groups. Total heterotrophic count, amylolytic, cellulolytic and proteolytic bacterial counts in gut were significantly (p < .01) higher in group II as compared to other two groups. Haematological analysis indicated better (p < .05) immune response in groups II and III than control group. Diets D2 and D3 showed better effect on growth, digestibility, FCR, survival and immune response of P. monodon as compared to D1.     

The extracts of Angelica sinensis restore the digestive and absorptive capacity through improving antioxidant status in digestive organs of fish treated with trichlorfon

Firstly, a linoleic acid emulsion and fish hepatopancreas homogenate were incubated with ethoxyquin and the extracts of Angelica sinensis. The results demonstrated that ethoxyquin showed the strongest protective effects against lipid oxidation of all of the examined compounds (p < 0.05). However, ethyl acetate extract of Angelica sinensis at high concentrations showed a stronger effect on lipid oxidation than that of ethoxyquin (p < 0.05). Next, seven experimental diets that contained 0.0, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 g/kg of ethyl acetate extract of Angelica sinensis were fed to seven groups of carp (Cyprinus carpio var. Jian) respectively. After 60 days, carp were exposed to 2.4 mg trichlorfon/L in water for 4 days. The results displayed that trichlorfon exposure increased the contents of malonaldehyde and protein carbonyl in digestive organs and the activities of glutamate‐oxaloacetate and glutamate‐pyruvate transaminase in plasma, and decreased feed intake, the level of reduced glutathione, and the activities of trypsin, chymotrypsin, lipase, alpha‐amylase, Na⁺,K⁺‐ATPase, alkaline phosphatase, antisuperoxide anion, antihydroxyl radical, superoxide dismutase, glutathione reductase, and glutathione S‐transferase in digestive organs of carp (p < 0.05). Moreover, the dietary ethyl acetate extract of Angelica sinensis prevented the decrease in the above parameters in carp treated with trichlorfon (p < 0.05). These results revealed that the dietary ethyl acetate extract of Angelica sinensis could quench the trichlorfon‐induced structural and functional damage by improving the antioxidative capacity of the digestive organs of fish. Therefore, the extract of Angelica sinensis could be used as an inhibitor of trichlorfon stress in fish.     

Effects of dietary coated protease on growth performance,feed utilization,nutrient apparent digestibility,intestinal and hepatopancreas structure in juvenile Gibel carp (Carassius auratus gibelio)

This study was conducted to investigate the effects of dietary protease on growth performance, feed utilization, whole‐body proximate composition, nutrient digestibility, intestinal and hepatopancreas structure of juvenile Gibel carp, Carassius auratus gibelio (mean weight 8.08 ± 0.18 g). Six diets were prepared, including a positive control diet (dietary protein 350 g/kg, PC), one negative control diet (dietary protein 33 g/kg, NC) and four protease supplementations diets, which were 75, 150, 300 and 600 mg/kg protease NC diet. After 12 weeks of diet feeding in indoor recycle aquarium tanks, no significant difference (p > .05) was found on growth performance between fish fed diet with 75–600 mg/kg protease and the PC group. Compared with the fish fed the NC diet, the specific growth rate of fish fed 300 mg/kg protease increased significantly (p < .05), as well as protein efficiency ratios (p < .05), while feed conversion was the opposite (p < .05). The nutrient digestibility of crude protein and lipid was higher (p < .05) in fish fed 150 mg/kg protease diet than the PC diet. Whole‐body proximate composition of fish was not affected (p > .05) by the dietary treatment. Serum alkaline phosphatase and albumin were significantly affected by dietary protease (p < .05), while the content of total protein, glucose, triglyceride, total cholesterol, aspartate aminotransferase and alanine aminotransferase activities in serum was not affected (p > .05). Foregut muscular thickness was thinner (p < .05), when the fish fed diets supplementation of protease in 150 or 600 mg/kg diet than the NC diet. Protease activities in hepatopancreas and foregut were higher (p < .05), in the fish fed 150 or 300 mg/kg protease diet than the fish fed the PC diet, but those in the mid‐ and hindgut were not significantly affected (p > .05) by the dietary treatments. Based on the regression analysis of weight gain rate, the optimal dietary inclusion level of protease was 400 mg/kg in the diet for juvenile Carassius auratus gibelio.     

Development of attenuated erythromycin‐resistant Streptococcus agalactiae vaccine for tilapia (Oreochromis niloticus) culture

Streptococcus agalactiae is an important pathogen in fish, causing great losses of intensive tilapia farming. To develop a potential live attenuated vaccine, a re‐attenuated S. agalactiae (named TFJ‐ery) was developed from a natural low‐virulence S. agalactiae strain TFJ0901 through selection of resistance to erythromycin. The biological characteristics, virulence, stability and the immunization protective efficacy to tilapia of TFJ‐ery were determined. The results indicated that TFJ‐ery grew at a slower rate than TFJ0901. The capsule thickness of TFJ‐ery was significantly less (p < 0.05) than TFJ0901. When Nile tilapia were intraperitoneally (IP) injected with TFJ‐ery, the mortality of fish was decreased than that injected with TFJ0901. The RPS of fish immunized with TFJ‐ery at a dose of 5.0 × 10⁷ CFU was 95.00%, 93.02% and 100.00% at 4, 8 and 16 weeks post‐vaccination, respectively. ELISA results showed that the vaccinated fish produced significantly higher (p < 0.05) antibody titres compared to those of control at 2 or 4 weeks post‐vaccination. Taken together, our results suggest that erythromycin could be used to attenuate S. agalactiae, and TFJ‐ery is a potent attenuated vaccine candidate to protect tilapia against S. agalactiae infections.     

Activities of digestive enzymes in relation to ingested natural food in three carp species of the Himalayan River Ladhiya,northern India

The present investigation was carried out to study gut content and accompanying digestive enzyme profiles of the three carp species, golden mahseer Tor putitora, snow trout Schizothorax richardsonii and hill trout Barilius barila. The fishes were captured from the River Ladhiya, Champawat district, Uttarakhand, northern India. The digestive tract length was on average 1.8 and 2.3 times longer than the body, while members of Bacillariophyceae, Chlorophyceae, Cyanophyceae and Zygnematophyceae made up 95.15% and 99.63% of gut content in the golden mahseer and snow trout, respectively. In hill trout, the digestive tract being only half the body length, gut content was insects (72.73%), eggs of aquatic organisms (22.73%) and nematodes (4.55%). Estimated gut enzyme activities demonstrated that amylase activity was significantly (P < 0.05) higher in golden mahseer and snow trout compared to hill trout, while total protease, trypsin and chymotrypsin activities were all significantly (P < 0.05) higher in the hill trout compared to other species. A strong co‐relation was found among relative gut length, food composition and digestive enzyme activities of each species. This may guide the future formulation of feed to these important species of carps kept in farm.     

The obvious heterosis and genetic characters of intergeneric cross and backcross juveniles between blunt snout bream (Megalobrama amblycephala) and topmouth culter (Culter alburnus)

In this study, we used artificial insemination to generate hybrid groups of fish [MC‐F₁(MA♀×CA♂) and MC‐F₂(MC‐F₁♀×♂)] by intergeneric crosses of Megalobrama amblycephala (MA) and Culter alburnus (CA); sequential backcrosses [CAM‐B₁ (CA♀×MC‐F₁♂) and MCC‐B₁ (MC‐F₁♀×CA♂)] were also performed. All these hybrids showed high rates of fertilization, hatching and survival (p > 0.05). For genetic traits, compared with those of the M. amblycephala and C. alburnus parental lines (Table 1), the fertilization rate, hatching rate and 7‐day survival rate of MC‐F1(MA♀×CA♂), MC‐F2(MC‐F1♀×♂), CAM‐B1 (CA♀×MC‐F1♂) and MCC‐B1 (MC‐F1♀×CA♂) by artificial insemination exhibited similar high rates (p > 0.05). The morphology of the four hybrids MC‐F₁/F_2, CAM‐B₁ and MCC‐B₁ were intermediate between those of their parents. Compared with their parents of MA and CA, weight gain rate (WG), specific growth rate (SGR) and protein efficiency ratio (PER) of hybrids MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ were significantly (p < 0.05) increased and feed conversion ratio (FCR) was significantly (p < 0.05) decreased after 3 months feeding. Moreover, protein content of muscle for MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ was significantly (p < 0.05) higher and carbohydrate content of muscle was significantly (p < 0.05) lower than their parents. The females and males of the four hybrids had normal gonadal development. In this study, we successfully generated intergeneric and backcross hybridization lines with fertile potential among fish of the Cultrinae subfamily and these hybrids had obvious heterosis in terms of growth performance, feed utilization and muscle quality.     

Determination of grapefruit (Citrus paradisi) peel extract bio‐active substances and its application in Caspian white fish (Rutilus frisii kutum) diet: Growth,haemato‐biochemical parameters and intestinal morphology

In this study, total phenolic and flavonoid contents of grapefruit peel extract (GPE) were equal to 117.3 ± 0.3 µg of gallic acid/mg and 39.30 ± 0.1 µg of quercetin/mg respectively. Caspian white fish (n = 180, 4 ± 0.9 g body weight) was fed with supplemented diets, including 0, 6.25, 12.5 and 25 mg of GPE/kg for 60 days at 25 ± 1°C. The growth performance was markedly improved in fish fed with 25 mg/kg of GPE compared to others (p < 0.05). Moreover, fish fed with 25 mg/kg of GPE showed a significant increase in red blood cell (2.65·10⁶ cell/mm³), white blood cell (17.75·10³ cell/mm³), packed cell volume (48%) and haemoglobin concentrations (8.75 g/dl) compared to the control (p < 0.05). However, the highest alanine aminotransferase (140 U/L), alanine transaminase (14.5 U/L), and alkaline phosphatase (18.5 U/L) were observed in control group. Morphological analysis of intestine revealed the highest amount of villus width (8.4 µM), height (32.86 µM) and surface area (342.7 µM²) in fish fed with 25 mg/kg of GPE (p < 0.05). In conclusion, supplementing feed with GPE at 25 g/kg can improve growth performance and haemato‐biochemical parameters of Caspian white fish fry.     

Effect of the dietary phosphatidylcholine at different growth stages of Pacific white shrimps,Litopenaeus vannamei

This study investigated the effects of dietary phosphatidylcholine (PC) at different growth stages of Litopenaeus vannamei (L. vannamei) with an initial weight of 0.70 ± 0.00 g (juvenile) and 4.25 ± 0.00 g (subadult) for 8 weeks. Six isonitrogenous and isoenergetic diets containing PC (0, 1.7, 5.1, 6.8, 10.2 and 13.6 g/kg) were confected using formulated with redfish meal, dehulled soybean meal and peanut meal as protein source, fish oil and corn oil as fat source. The juveniles were reared in 0.3 m³ aquarium at the density of 40 in triplicates, as well as the subadults in 0.5 m³ at the density of 40 in triplicates. Results showed PC level had significant effects on weight gain rate (WGR), specific growth rate, feed conversion ratio in juvenile and subadult shrimps and hepatosomatic index in juvenile shrimps (p < 0.05). No significant differences were found in moisture, whole body protein and crude ash (p > 0.05). Crude fat of juveniles had no significant change (p > 0.05), but subadults increased significantly (p < 0.05). Both juveniles and subadults showed that triglyceride and low‐density lipoprotein presented a downward trend with PC elevating, but high‐density lipoprotein increased gradually. Cholesterol in juveniles presented upward, but downward in subadults (p < 0.05). Dietary PC also significantly reduced the aspartate transaminase in juveniles and alanine transaminase in subadults (p < 0.05). PC supplementation significantly improved the glutathione S‐transferase activity and reduced the content of malondialdehyde in the hepatopancreas (p < 0.05). PC supplementation significantly improved gill filament sodium‐potassium adenosine triphosphatase enzyme of juvenile shrimps (p < 0.05), but subadult shrimps were not influenced (p > 0.05). PC supplementation significantly elevated the content of hepatopancreas eicosapentaenoic acid, highly unsaturated fatty acid (HUFA) and n‐3HUFA (p < 0.05), but the content of linoleic acid significantly reduced (p < 0.05). The broken‐line and quadratic regression model based on WGR showed the optimum dietary PC requirement was 10.3 g/kg and 6.5 g/kg for juveniles and subadults, respectively.     

Microbial communities associated with early stages of intensively reared orange‐spotted grouper (Epinephelus coioides)

The gut microbiota plays key roles in the health and general welfare of fish larvae, the present study characterized the bacterial communities associated with grouper Epinephelus coioides larvae during a period of 22 days post hatch (DPH) in an intensive hatchery using both cultivation‐based and cultivation‐independent approaches. Both approaches confirmed that bacteria were present in the gut of larvae before and after the onset of exogenous feeding, and the number of cultiviable bacteria increased gradually from 2 DPH to 22 DPH. A more complex bacterial profile was present in larvae fed fertilizer oyster eggs for 4 days (8 DPH), probably as a result of the onset of exogenous feeding. Interestingly, similar internal microbiota were observed in larvae fed fertilized oyster eggs for 4 days (8 DPH) and rotifers for 2 weeks (22 DPH), although different microbial communities were present in the two feeds. This might suggest that the gut environment of E. coioides larvae selects for a common microbiota, which is more closely related with the rearing water than the two feeds. Therefore, bacterial community of the rearing water may play a critical role in the establishment of gut microbiota of fish larvae and more attention should be paid to its practical modulation by using probiotics. In addition, some potentially beneficial bacteria, such as Lactococcus spp., were the major components of the microbiota associated with fertilized oyster eggs, while these bacteria were not detected in larvae samples.     

Oral administration of Escherichia coli lipopolysaccharide enhances the immune system of striped catfish,Pangasianodon hypophthalmus (Sauvage)

Lipopolysaccharide (LPS), a component of Gram negative bacteria, was reported as important immunostimulant for fish. In this study, striped catfish were fed diets containing different Escherichia coli LPS concentrations (0%, 0.01% and 0.05%) for 2 weeks and then fed control feed (0% LPS) for 4 weeks. Plasma cortisol and glucose were rather low and did not differ significantly among treatments (P > 0.05). The respiratory burst activity, lysozyme, complement, total of antibody as well as mortality in fish challenged with Edwardsiella ictaluri were recorded every 2 weeks (W2, W4 and W6). The lysozyme activity significantly increased in fish treated with LPS (P < 0.05) in W2, W4 and W6. The highest values of respiratory burst activity were observed at week 4 in fish fed 0.01% LPS. There were significant differences in total of antibody between fish fed LPS (0.01%) and control in W2, W4. The challenge test with Edwardsiella ictaluri showed that fish fed 0.01% LPS had lower cumulative mortality (40%, 33% and 42%) compared with the fish fed 0.05% LPS (50%, 40% and 47%) and control fish (40%, 57% and 53%) in the three difference sampling times respectively. These results suggest that feed supplemented with 0.01% LPS could enhance immunity of striped catfish after 2 weeks of oral administration and fish could be protected against bacterial infection during the following 4 weeks.     

Effects of replacing fish meal with corn gluten meal on growth,feed utilization,nitrogen and phosphorus excretion and IGF‐I gene expression of juvenile Pseudobagrus ussuriensis

Seven isonitrogenous and isolipidic diets containing fish meal (FM) protein replaced by corn gluten meal (CGM) protein at 0% (the control, C0), 10% (C10), 20% (C20), 30% (C30), 40% (C40), 50% (C50) and 60% (C60) were fed to juvenile Pseudobagrus ussuriensis for 8‐weeks to evaluate the effects of FM protein replaced by CGM protein on growth, feed utilization, nitrogen (N) and phosphorus (P) excretion and IGF‐I gene expression of juvenile P. ussuriensis. The results showed that the replacement level up to 40% did not affect the weight gain, specific growth rate (SGR), feed intake and protein efficiency ratio, whereas these parameters were depressed by further replacement level. Apparent digestibility coefficients (ADC) of dry matter, crude protein significantly decreased, but ADC of phosphorus significantly increased with increasing dietary CGM levels (p < .05). Fish fed diets with FM protein replaced by CGM protein led to an increase in nitrogen excretion, but led to a reduction in phosphorus excretion. No significant differences were observed in alpha‐amylase and lipase activities of intestine (p > .05). The lowest pepsin activity was found in C60 group. Fish fed diet C40, C50 and C60 had significantly lower serum lysozyme activity compared with fish fed diet C0 (p < .05). The lowest plasma alkaline phosphatase activity and the highest plasma alanine aminotransferase and aspartate aminotransferase activities were observed in C60 group. Fish fed diet C60 had significantly lower hepatic IGF‐I gene expression compared with fish fed diet C10 (p < .05). Broken‐line model analysis based on SGR against the CGM substitution level indicated that the appropriate replacement level was 37.7%.