Effects of dietary fish oil replacement with blended vegetable oils on growth,lipid metabolism and antioxidant capacity of subadult swimming crab Portunus trituberculatus

A 120‐day trial was conducted to assess the effects of dietary fish oil replacement with vegetable oils on growth, lipid metabolism and antioxidant capacity of subadult swimming crab Portunus trituberculatus. Five isonitrogenous and isolipidic diets were formulated to replace 0, 250, 500, 750 and 1000g/kg of fish oil with a mixture of soybean and rapeseed oil (defined as D1–D5), and each treatment had 30 replicate crabs. Dietary fish oil replacement had no significant effects on growth of the crabs, while the D3 had the highest hepatosomatic index and total lipids in hepatopancreas. The triglyceride and lipase activities in hepatopancreas increased significantly with increasing dietary fish oil replacement. The D4 had the highest levels of alkaline phosphatase (ALP) and acid phosphatase (ACP) in the hepatopancreas, as well as the haemolymph ALP, ACP and peroxidase. The highest levels of haemolymph total antioxidant capacity, catalase and malondialdehyde were detected in D1. Total n‐3 polyunsaturated fatty acids levels in hepatopancreas decreased significantly with increasing dietary fish oil replacement. In conclusion, dietary fish oil replacement had no significant effects on growth of P. trituberculatus, and 500g/kg of fish oil replacement could improve antioxidant capacity, but excessive replacement level will enhance lipid accumulation and peroxidation in the hepatopancreas.     

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 astaxanthin on the growth,innate immunity and antioxidant defence system of Paramisgurnus dabryanus

The effects of astaxanthin supplementationon the growth, innate immunity and antioxidant defence system of loach (Paramisgurnus dabryanus) were investigated in this study. A total of 450 fish (initial average weights of 3.00 ± 0.10 g) were fed five diets with graded levels of astaxanthin (0.00, 50.00, 100.00, 150.00 and 200.00 mg/kg) for 56 days. The results showed that astaxanthin supplementation significantly stimulated the growth (FBW, WG, FER, HSI), innate immunity (AKP activity in the hepatopancreas, intestinal tract, muscle and skin and LZM activity in the hepatopancreas, intestinal tract and skin), antioxidant ability (T‐SOD, GSH‐PX and CAT activities and T‐AOC, GSH and MDA contents in the hepatopancreas, intestinal tract, muscle and skin) of loach. Moreover, dietary astaxanthin supplementation significantly up‐regulated the relative mRNA levels of Nrf2 and Maf, and down‐regulated the relative mRNA level of Keap1 in the hepatopancreas when the supplementation levels of astaxanthin were 50–200 mg/kg. In conclusion, the diets with 100–151.06 mg/kg astaxanthin supplementation were optimal for loach, which based on the growth, immunity and antioxidant‐related indicators, and the astaxanthin supplementation regulated the antioxidant ability partially referring to Keap1‐Nrf2 signallings.     

Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process

We have developed a two-stage growth one-step process for cultivation of Haematococcus using a self-designed system that mimics an open pond in the natural environment. The characteristics of this process are green vegetative cell growth and cysts transformation and pigment accumulation that proceed spontaneously and successively in one open photobioreactor. Four strains of Haematococcus (H. pluvialis 26; H. pluvialis 30; H. pluvialis 34; H. pluvialis WZ) were cultured in this imitation system for a duration of 12 days. The changes in cell density and medium pH were closely monitored, and the astaxanthin content and yield of the four Haematococcus strains were measured at the end of 12 days of cultivation. Two of the strains, H. pluvialis 26 and H. pluvialis WZ, were selected as strains suitable for mass culture, resulting in the astaxanthin yield of 51.06 and 40.25 mg L^− 1 which are equivalent to 2.79 and 2.50% of their dry biomass respectively. Based on the laboratory work, 6 batch cultures of H. pluvialis WZ were conducted successfully to produce astaxanthin in two 100 m² open race-way pond by two-stage growth one-step process. The astaxanthin content ranged from 1.61 to 2.48 g 100 g^− 1dry wt., with average astaxanthin content of 2.10 g 100 g^− 1dry wt. Compared with the one-stage production of astaxanthin based on continuous culture, the superiority of our process is that it can accumulate much more astaxanthin in red cysts. Compared with two-stage production of astaxanthin, the advantage of our process is that it does not need to divide the production process into two parts using two bioreactors. The presented work demonstrates the feasibility for producing astaxanthin from Haematococcus using a two-stage growth one-step process in open pond, culture systems that have been successfully used for Spirulina and Chlorella mass culture. The future of Haematococcus astaxanthin production has been also discussed.     

Changes of carotenoids contents and analysis of astaxanthin geometrical isomerization in Haematococcus pluvialis under outdoor high light conditions

In this study, carotenoid contents of Haematococcus pluvialis during outdoor high light cultivation were measured, moreover, changes of astaxanthin geometrical isomers and biotic factors which may affect isomerization were investigated. During the incubation, contents of both astaxanthin and its precursors (zeaxanthin and canthaxanthin) increased over time, whereas the relative content of lutein decreased. Contents of all astaxanthin isomers increased, while the proportion of different astaxanthin geometrical isomers fluctuated during the incubation. All‐trans‐astaxanthin of total astaxanthin (T/A) was higher during the astaxanthin accumulation phase than that in the cell transformation phase, which was in contrast to results of 13‐cis‐astaxanthin of total astaxanthin (13C/A) and 9‐cis‐astaxanthin of total astaxanthin (9C/A). Farnesyl diphosphate synthase (trans (2E,6E)‐FPPS, EC2.5.1.10) and geranylgeranyl diphosphate synthase (GGPS, EC2.5.1.29), the key proteins involved in geometrical isomerization, decreased during the astaxanthin accumulation phase. Moreover, the presence of cis (2Z,6E)‐FPPS was firstly confirmed in H. pluvialis by HPLC‐MS/MS shotgun method. The results indicated the biotic factor (trans‐FPPS, cis‐FPPS and GGPS) may play an important role in astaxanthin geometrical isomerization of H. pluvialis, but not a crucial role. This study would help in optimizing the regulation of astaxanthin geometrical isomers in H. pluvialis, with great significance in theory and production.     

Effects of dietary vitamin E and astaxanthin on growth,skin colour and antioxidative capacity of large yellow croaker Larimichthys crocea

A 10‐week feeding trial was conducted to evaluate the effect of dietary vitamin E and astaxanthin on growth performance, skin colour and antioxidative capacity of large yellow croaker Larimichthys crocea. Six practical diets were formulated in a 2 × 3 factorial design to supplement with two levels of astaxanthin (25 and 50 mg/kg) and three levels of vitamin E (0, 120 and 800 mg/kg). The results showed that both the highest final body weight and specific growth rate were found in fish fed diets with 120 mg/kg vitamin E supplementation. No significant differences were found in survival rate, feed conversion ratio and protein efficiency ratio among all the treatments (p > .05). Skin lightness (L*) was not significantly affected by dietary treatments (p > .05). Ventral skin redness (a*) of fish fed diet with 25 mg/kg astaxanthin and 0 mg/kg vitamin E supplementation was significantly lower than that of fish fed with other diets. Yellowness (b*) and carotenoid contents both in the dorsal and in the ventral skin were found to be significantly increased with increasing dietary astaxanthin or vitamin E (p < .05), but no significant interactions were found (p > .05). The vitamin E content in liver reflected the dietary vitamin E content. Level of vitamin E content in fish fed diets with 800 mg/kg vitamin E supplementation was significantly higher than that in fish fed with the other diets (p < .05). Liver superoxide dismutase activity and thiobarbituric acid reactive substance levels were found to be decreased with increasing dietary astaxanthin and vitamin E levels, respectively. Levels of reduced glutathione in the liver were found to be increased with increasing dietary vitamin E contents. The total antioxidative capacity in the liver was found to be decreased with increasing dietary vitamin E or astaxanthin contents. In conclusion, adequate dietary vitamin E can improve the growth of large yellow croaker, and the supplementation of astaxanthin and vitamin E benefited the skin coloration and antioxidative capacity of large yellow croaker.     

Effect of dietary leucine on growth performance,hemolymph and hepatopancreas enzyme activities of swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to determine the dietary leucine requirement for juvenile swimming crabs reared in cement pools. Six isonitrogenous and isolipidic practical diets (430 g/kg crude protein and 70 g/kg crude lipid) were formulated to contain graded leucine levels which ranged from 16.7 to 26.7 g/kg (dry weight). Each diet was randomly assigned to triplicate groups of 60 juvenile swimming crabs (initial average weight 3.75 ± 0.12 g) that were stocked in rectangle plastic baskets. The results of the present study indicated that dietary leucine levels significantly influenced weight gain (WG) and specific growth ratio (SGR) (p < .05), crab fed the diet containing 22.7 g/kg leucine had significantly higher WG and SGR than those fed the other diets. Feed efficiency and protein efficiency ratio were not significantly affected by the dietary leucine levels (p > .05). Total protein, cholesterol, triglyceride and glucose in serum were significantly affected by the dietary leucine levels. Aspartate aminotransferase (AST) and alanine aminotransferase activities in hemolymph, AST and superoxide dismutase activities in hepatopancreas were significantly affected by dietary leucine levels; moreover, crab fed the 16.7 g/kg leucine diet had higher malondialdehyde in hemolymph and hepatopancreas than those fed the other diets. Crab fed the diet containing 24.9 g/kg leucine had higher phenoloxidase activity in hemolymph than those fed the other diets. Based on two‐slope broken‐line model of SGR against dietary leucine levels, the optimal dietary leucine requirement for growth was estimated to be 22.1 g/kg of the dry diet (corresponding to 51.4 g/kg of dietary protein on a dry weight basis). In summary, findings of this study indicated that dietary leucine could improve growth performance and antioxidant status.     

Effects of Chinese herbal medicines mixture on growth performance digestive enzyme activity immune response of juvenile Japanese seabass,Lateolabrax japonicus

A single factorial experiment was conducted to investigate the effects of Chinese herbal medicines mixture (CHMM) on growth performance, digestive enzyme activity and immune response of Japanese seabass, Lateolabrax japonicus (initial weight 5.01 ± 0.32 g). The fish were fed diets containing six levels of CHMM (0, 4, 8, 12, 16 and 20 g/kg) for 4 weeks. The results showed that the weight gain rate and specific growth rate (SGR) enhanced significantly in fish fed diet containing 8 g/kg CHMM (p < .05), while the feed conversion ratio (FCR) in the 4 and 8 g/kg CHMM groups reduced significantly compared with the control (p < .05). The body crude protein levels in 8 and 16 g/kg groups were significantly higher than the control (p < .05). The pepsin, amylase and lipase activities elevated significantly in the stomach of fish fed 8 g/kg CHMM, while the erepsin and lipase in the intestine of fish in 12 and 16 g/kg groups were higher significantly than the control (p < .05). The lysozyme, alkaline phosphatase, acid phosphatase (ACP), total antioxidative capacity activities in serum of fish fed 12 g/kg CHMM were higher significantly than those in the control (p < .05), while the total superoxide dismutase (SOD), total antioxidative capacity, catalase, alkaline phosphatase (AKP) and ACP activities in hepatopancreas of fish in 12 g/kg group were all significantly higher than those in the control (p < .05). Regression analysis showed that the relationships between dietary CHMM levels and either FCR, SGR, erepsin, pepsin or lysozyme activities were best expressed by quadratic or cubic regression equations, and the optimal inclusion levels are 11.4, 10.7, 10.7, 8.4 and 10.5 g/kg for maximum FCR, SGR, erepsin, pepsin and lysozyme activities, respectively. Under the present experimental condition, the optimal supplementary level of CHMM in the diet of Japanese seabass is 8–12 g/kg.     

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.     

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.     

Effect of lactic acid on enrofloxacin pharmacokinetics in Eriocheir sinensis (Chinese mitten crab)

As a representative acidifier, lactic acid (LA) is widely used in the diets of aquatic animals. LA is the main supporter of energy metabolism and may be associated with drug metabolism. This study was designed to explore the pharmacokinetics of enrofloxacin (ENR) in Eriocheir sinensis (Chinese mitten crab) fed diets containing 0.1%, 0.2% and 0.3% LA (designated groups LA1, LA2 and LA3 respectively). The concentrations of ENR in the haemolymph, hepatopancreas and muscle were determined by HPLC after a single oral dose of 10 mg/kg. Our results showed that LA had a significant effect on the peak ENR concentrations in all the tissues (p < 0.05) by one‐way ANOVA analysis. There was a trend that C_max (peak concentration) of ENR was elevated with LA levels increasing up to 0.3% in haemolymph, hepatopancreas, muscle and T_max (time‐point of the peak concentration of the drug), t_1/2β (elimination half‐life) and AUC_(0‐∞) of ENR were shortened. Taken together, 0.3% LA might be effective in improving ENR pharmacokinetics in E. sinensis. Furthermore, it can be speculated that the enhanced biotransformation of ENR in the hepatopancreas mediated by LA is responsible for the differences in the pharmacokinetics of ENR in E. sinensis.     

Effects of dietary astaxanthin on growth,blood biochemistry,antioxidant, immune and inflammatory response in lipopolysaccharide‐challenged Channa argus

The present study investigated the effects of dietary astaxanthin on the growth, blood biochemical, antioxidant, immune and inflammatory response in lipopolysaccharide‐challenged Channa argus. A total of 0, 50, 100 and 200 mg/kg of astaxanthin were added to the basal die for 56 days. After the feeding experiment, each group was subjected to a lipopolysaccharide challenge (except for the Control group). The results showed that adding astaxanthin to the diet can significantly increase the weight gain and specific growth rate and decrease the feed conversion ratio of C. argus; the highest weight gain, specific growth rate and minimum feed conversion ratio occurred in the 100 mg/kg group. Furthermore, dietary astaxanthin supplementation can alleviate the negative effects of lipopolysaccharides by increasing the levels of alkaline phosphatase, lysozyme, complement 3, complement 4, total serum protein, albumin, globulin, superoxide dismutase, catalase and glutathione peroxidase and decrease the serum cortisol, aspartate aminotransferase, alanine aminotransferase, glutathione peroxidase, and malondialdehyde. Dietary astaxanthin supplementation also can decrease the relative expression of inflammatory genes (nuclear factor κB, interleukin‐1, interleukin‐8 and tumour necrosis factor‐α) in the liver, spleen, kidney and intestine. To summarise, dietary astaxanthin addition can improve the growth performance and attenuate the negative effects of lipopolysaccharide challenge in C. argus. The optimal amount of astaxanthin is 100 mg/kg.     

Hypoxia‐induced changes in survival,immune response and antioxidant status of the Pacific white shrimp (Litopenaeus vannamei) fed with graded levels of dietary myo‐inositol

A 3‐hr experiment was conducted to investigate the effects of dietary myo‐inositol (MI) supplementation on survival, immune response and antioxidant abilities in Litopenaeus vannamei under acute hypoxia stress. Six practical diets were formulated with supplementation of graded levels (control group 0, 0.1, 0.2, 0.4, 0.8 and 1.6 g/kg dry diet) of MI and were randomly assigned to triplicate groups of L. vannamei (mean weight 0.40 ± 0.00 g) for 8 weeks. Ten healthy shrimp (final mean weight approximately 11–14 g) randomly selected from each tank were exposed to hypoxia stress after feeding trial. After 3‐hr acute hypoxia stress, survival of shrimp fed MI‐supplemented diets (except 0.1 and 0.4 g/kg diets) was significantly increased compared with the control group. Shrimp fed control diet had lower activities of alkaline phosphatase (AKP), acid phosphatase (ACP), total antioxidant capacity (T‐AOC) and glutathione peroxidase (GPX), and higher malondialdehyde (MDA) and protein carbonyl (PC) contents in hepatopancreas than those fed the MI‐supplemented diets. In addition, mRNA expression levels of heat shock protein 70 (Hsp70), catalase (CAT) and penaeidin were significantly differentially regulated in hepatopancreas. In summary, dietary MI supplementation may have a positive effect on improving resistance to acute hypoxia stress of L. vannamei.     

Effects of fishmeal replacement with cottonseed meal protein hydrolysate on growth,digestion and intestinal histology of juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

A 60‐day feeding experiment was conducted to evaluate the effects of fishmeal (FM) replacement with cottonseed meal protein hydrolysate (CPH) on growth, digestion and intestinal histology of juvenile Chinese soft‐shelled turtle (Pelodiscus sinensis). Five diets were formulated to replace 0, 50, 100, and 150 g/kg fishmeal protein by CPH (CPH0, CPH5, CPH10, CPH15) and CPH15L (CPH15 with micro capsule‐L‐lysine). Weight gain, feed conversion rate and protein efficiency ratio showed no significant differences compared to control group (p > .05). The highest feed intake indicated in CPH15 (p < .05). The composition of whole‐body varied slightly in each groups (p > .05). The trypsin activity significantly elevated when dietary fishmeal protein was replaced by CPH at 30–90 g/kg (p < .05). A significantly higher lipase activities in CPH5 than control group (p < .05). The CPH5‐10 groups showed higher villus height than the other groups (p < .05). The microvillus length in turtles with CPH showed a significant increasing length (p < .05). The results indicated that replacing up to 90 g/kg of dietary fishmeal protein with CPH did not hamper growth or reduce feed intake of turtles. Moreover, CPH replaced 60 g/kg FMP can increase intestine digestive enzymes activities and improve intestinal development.     

Effects of γ‐aminobutyric acid supplementation on the growth performance,serum biochemical indices and antioxidant status of pharaoh cuttlefish,Sepia pharaonis

This study evaluated the effects of dietary γ‐aminobutyric acid (GABA) on the growth performance, serum biochemical indices and antioxidant status of pharaoh cuttlefish, Sepia pharaonis. Cuttlefish were cultured in open‐culturing cement pool systems for 8 weeks. Six practical diets supplemented with graded levels of GABA (0, 20, 40, 60, 80 and 100 mg/kg) were formulated. Each diet was randomly assigned to triplicate groups of 60 cuttlefish (mean weight: 10.33 g), the cuttlefish were fed two times per day to apparent satiation. The results showed that the specific growth rate (SGR), weight gain (WG) and feed efficiency (FE) significantly increased with dietary GABA supplementation (p < .05). The survival rate (SR) and protein content in muscle significantly increased when 58.9 mg/kg GABA supplied. Moreover, the nitric oxide (NO) content and acid phosphatase (ACP) activity in serum were significantly increased with dietary GABA supplementation (p < .05), while the activity of aspartate aminotransferase (AST) in serum decreased significantly when supplied with GABA at 58.9 mg/kg (p < .05). In addition, dietary GABA improved antioxidation activity by significantly increasing the activities of superoxide dismutase (SOD) and catalase (CAT) but decreasing malondialdehyde (MDA) levels in the liver and gill (p < .05). On the basis of the quadratic regression analysis of FE, the optimum content of dietary GABA in S. pharaonis was estimated to be 55.3 mg/kg. The findings of this study demonstrated that dietary GABA had a positive effect on the growth performance, serum biochemical indices and antioxidant status of S. pharaonis.     

Effects of different dietary vitamin E and astaxanthin levels on growth,fatty acid composition and coloration of the juvenile swimming crab,Portunus trituberculatus

Six isonitrogenous and isolipidic diets were formulated containing two astaxanthin (AX; 0 and 90 mg/kg) and three vitamin E (VE) levels (0, 25 and 50 mg/kg). There were three replicates (18 crabs per replicate) for each treatment. Juvenile swimming crab (initial weight 31.65 ± 0.06 g/crab) were fed different diets for 8 weeks. After the feeding trial, growth performance was not significantly affected by the different treatments. Crabs fed with AX‐supplemented diets showed more redness. Whole body 22:6n‐3, 22:4n‐6 and 20:5n‐3 levels increased with the dietary addition of AX (P = .009, P = .002 and p = .042, respectively). The malondialdehyde (MDA) concentrations of fresh/frozen hepatopancreas and frozen muscle were significantly decreased by the dietary AX supplementation (p < .001, p = .010 and p = 0.011, respectively). These findings provide evidence that dietary AX has an ability to improve the redness of the shell and reduce the MDA concentrations of tissues. Furthermore, there is no strong interactive relationship between dietary VE and AX on the coloration and fatty acid concentrations for the swimming crab.     

Development of In situ hybridization and real‐time PCR assays for the detection of Hepatospora eriocheir,a microsporidian pathogen in the Chinese mitten crab Eriocheir sinensis

A microsporidian parasite, Hepatospora eriocheir, is an emerging pathogen for the Chinese mitten crab Eriocheir sinensis. Currently, there is scant information about the way it transmits infection in the crustacean of commercial importance, including its pathogenesis, propagation and infection route in vivo. In this study, chromogenic in situ hybridization (ISH) and quantitative real‐time PCR (qPCR) assays were developed to address this pressing need, and we provided an advance in the detection methods available. Pathogens can be seen in situ with associated lesions using ISH. Positive hybridization signals were noted inside the epithelial cells of the hepatopancreas, and putative free parasite spores were observed within the tubule lumen, which were associated with lesions detected by electron microscopy and haematoxylin and eosin (H&E) analysis. qPCR allows the determination of parasite loads in infected tissues, which is important for understanding disease progression and transmission. The hepatopancreas displayed the biggest statistical copy numbers among different tissues of infected crabs, confirming a tissue‐specific pathogen infection characteristic. The qPCR assay also proved to be suitable for the diagnosis of asymptomatic carrier crabs. Combination of the two methods could facilitate the study of H. eriocheir infection mechanism in E. sinensis, enhance the early diagnosis of the pathogen and improve the management of microsporidian diseases in commercial crustaceans.     

Effects of different dietary lipid sources on growth performance,antioxidant enzyme activities and biochemical composition of juvenile swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to evaluate the effects of dietary lipid sources on growth performance, antioxidant enzyme activities and biochemical composition of juvenile swimming crab Portunus trituberculatus of initial weight 2.34 ± 0.08 g. Four different diets were formulated to contain fish oil (FO), soybean oil (SO), linseed oil (LO) or palm oil (PO). The highest final body weight, weight gain, specific growth rate and molting frequency were observed in crabs fed the FO diet. Crabs fed the SO diet showed higher glutathione peroxidase, superoxide dismutase (SOD) and catalase (CAT) activities in both serum and hepatopancreas than those fed the FO diet. The lowest malondialdehyde concentration in hepatopancreas and serum were occurred at crabs fed the SO diet. Crabs fed the LO diet had significantly higher SOD and CAT activities in hepatopancreas compared with those fed the FO diet. Crabs fed the PO diet had the highest activities of fatty acid synthase and carnitine palmitoyltransferase 1 activities in hepatopancreas among all treatments. Fatty acid compositions both in hepatopancreas and muscle reflected those of diets. Overall, these findings demonstrated that physiological–biochemical characteristics and lipid metabolism were significantly regulated by different dietary lipid sources. Moreover, dietary SO and LO supplementation could improve antioxidant ability.