Dietary krill hydrolysates affect the expression of growth‐related genes in juvenile turbot (Scophthalmus maximus L.)

A 10‐week feeding experiment in indoor flow‐through seawater system was conducted to investigate the effects of dietary krill hydrolysate on the expression of growth‐related genes in juvenile turbot (Scophthalmus maximus L.; initial body weight 9.45 ± 0.01 g). Three isonitrogenous and isolipidic experimental diets containing high plant protein were formulated to contain 0 (control), 50 g/kg (LKH) and 100 g/kg (HKH) krill protein hydrolysate (KH) to replace fishmeal, respectively. Triplicate groups of 30 fish were fed for 10 weeks to apparent satiation twice daily. At the end of the feeding trial, the mRNA expressions of insulin‐like growth factor (IGF‐1) gene in liver, peptide transporters (PepT1) gene in pyloric caeca and proximal intestine and neuropeptide Y (NPY) gene in brain in all groups were determined. IGF‐1, PepT1 and NPY expression levels in HKH group were significantly increased compared with those of LKH and control (p < 0.05), which was consistent with the SGR, feed efficiency, PER and PPV. These results indicated that dietary 100 g/kg krill hydrolysate could improve growth performance and upregulate the mRNA expression of IGF‐1, PepT1 and NPY genes in juvenile turbot.     

Edwardsiellosis in farmed turbot,Scophthalmus maximus (L.), associated with an unusual variant of Edwardsiella tarda: a clinical,aetiological and histopathological study

During 2005 and 2010, a survey of edwardsiellosis on eight turbot, Scophthalmus maximus (L.), farms was conducted in China. This report presents the detailed results of the study on this disease. Diseased turbot displayed two distinct types of gross signs: black discoloration of the dorsal skin on the posterior portion of the body; and red cutaneous foci on the ventral side. Internally, the most pronounced clinical signs in all fish examined were enlarged kidneys. The causal agent of the disease was finally proved to be one species of bacterium that was identified as Edwardsiella tarda by physiological and biochemical tests, API 32E and 16S ribosomal RNA sequence analysis. It is noteworthy that unlike the commonly described E. tarda strains, the isolates in this study were non‐motile strains without flagella. A histopathological study revealed that E. tarda infection was systemic in turbot and that kidney showed the most significant pathological changes, including acute focal necrosis, an influx of macrophages and formation of granuloma. The most common histopathological characteristics of this disease are the proliferation of macrophage in various organs and formation of granuloma. In addition, this article also gave background information on the disease and presented the results of virulence tests with the E. tarda strain identified in this study.     

Fish protein hydrolysate in diets of turbot affects muscle fibre morphometry,and the expression of muscle growth‐related genes

The study was conducted to investigate the effects of fish protein hydrolysate (FPH) in diets for turbot on growth performance, muscle fibre morphometry, and the expression of muscle growth‐related genes. A control diet (FPH0) contained 0 g/kg FPH, and four experimental diets were formulated replacing fishmeal with FPH at levels of 45 (FPH4.5), 120 (FPH12), 180 (FPH18) and 300 (FPH30) g/kg. Fish fed the FPH12 and FPH18 diets had higher specific growth rate (SGR) than fish fed the FPH0 diet (p < .05), and a quadratic regression was found between SGR and dietary FPH level (p = .001, R² = .677). Cross‐section area (CSA) and the length of sarcomere in the FPH12 group increased compared with the control group (p < .05), and a quadratic regression was observed between CSA and dietary FPH level (p = .006, R² = .574) and between sarcomere length and dietary FPH level (p = .018, R² = .788). An appropriate level of FPH down‐regulated myostatin 2 gene expression and up‐regulated proliferating cell nuclear antigen gene expression, while the expression of myogenic regulatory factors was not affected by dietary treatments (p > .05). To conclude, an appropriate level of FPH may improve muscle growth by regulating the expression of muscle growth‐related genes, and muscle microstructure and ultrastructure.     

Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

Differential expression of lipid metabolism‐related genes and miRNAs in Ctenopharyngodon idella liver in relation to fatty liver induced by high non‐protein energy diets

To explore the mechanism of fatty liver formation induced by high non‐protein energy diets in grass carp (Ctenopharyngodon idella), basal diet and high‐energy diets were fed to juvenile grass carp for 9 weeks. The experimental groups fed on high‐energy diets which included a high‐lipid diet (H‐LIP), a high‐carbohydrate diet (H‐CHO) and a high‐lipid and carbohydrate diet (H‐CL). The control group fed on basal diet. Growth performance, liver fat accumulation, serum biochemical indexes and the expression levels of lipid metabolism‐related genes (SREBP‐1, PPARγ, FAS, ACC1, and LPL) and miRNAs (miR‐33, miR‐122, and miR‐370) were examined at the end of the feeding trial. There were no significant differences in growth rate and feed efficiency among the four groups. However, significant increase in mesenteric and liver fat contents, and lipid droplets in the liver was induced by high‐lipid and high‐carbohydrate diets. There were significant differences in serum biochemical indicators such as AST/ALT, GLB, TG and TP, and liver fatty acid composition between the control and experimental groups. The expression levels of SREBP‐1, PPARγ, FAS, ACC1 and LPL were upregulated, while CPT‐1 was downregulated with the high‐energy treatments. Additionally, the expression levels of miR‐33, miR‐122 and miR‐370 in the liver were higher in the three high‐energy treatments than those in the control (P < 0.05). The results suggest that modifications of lipid metabolism‐related genes and miRNAs may be involved in fatty liver formation induced by high non‐protein energy diets in grass carp.     

Genotyping‐by‐sequencing for construction of a new genetic linkage map and QTL analysis of growth‐related traits in Pacific bluefin tuna

Pacific bluefin tuna (Thunnus orientalis) has high market value, but its wild populations have decreased in recent years. The broodstock of Pacific bluefin tuna that were hatched artificially and reared under aquaculture conditions is beginning to be used for production. The creation of broodstock with commercially valuable traits, such as rapid growth, is therefore of great interest. Genetic linkage map‐based identification of markers associated with quantitative trait loci (QTLs) facilitates marker‐assisted selection (MAS) breeding and allows efficient genetic improvement of broodstock. Single nucleotide polymorphism (SNP)‐based genetic linkage map construction using the genotyping‐by‐sequencing method can expand the number of mapped markers and help identify growth‐related QTLs. In this study, we constructed sex‐specific maps for 24 linkage groups consisting of 677 SNP and 651 microsatellite markers. The total lengths of 93 progenies in the mapping population followed normal distribution, with an average length of 9.4 mm. We performed composite interval mapping in the mapping population. QTL analysis revealed one significant QTL in LG10 on the female linkage map. The genetic linkage map—the second such map generated for Pacific bluefin tuna—and the growth‐related QTLs detected in this study will be useful for tuna aquaculture MAS programs.     

Comparative genomic insights into the taxonomy of Edwardsiella tarda isolated from different hosts: Marine,freshwater and migratory fish

Edwardsiella tarda, a Gram‐negative member of the family Enterobacteriaceae, has been isolated from many animal species worldwide, especially fish species. Its broad host range indicates the diversity in taxonomy, which attracted the attention of many researchers. Here, we added genome of E. tarda strain isolated from freshwater fish to comparative genomics study for the first time. We sequenced and assembled the genome of E. tarda ASE201307 which was isolated from freshwater Asian swamp eel. ASE201307 genome contained a single circular chromosome of 3.68M with G+C 57.09% content. Comparative genomics including SNP calling, synteny block, Core/Pan genes analysis and phylogeny analysis was conducted among ASE201307 and other Edwardsiella strains isolated from different fish species. Results of SNP analysis and synteny block demonstrated the close relative of ASE201307, FL95.01 and DT which were all isolated from freshwater fish. In further analysis heat map of dispensable genes and phylogenetic tree, all E. tarda strains were divided into two groups. One was isolated from freshwater fish and the other was isolated from marine/migratory fish. Based on all studies above, we proposed the living environment of hosts as a new taxonomic character and divided E. tarda isolated from diseased fish into freshwater group and marine/migratory group.     

Non‐invasive fast real‐time PCR assay for detection of the enteric parasite Enteromyxum scophthalmi in cultured turbot (Scophthalmus maximus L.)

Enteromyxum scophthalmi is a myxozoan parasite that causes severe parasitic diseases in cultured turbot affecting mainly the intestine of the host. It is characterized by producing acute enteritis, starvation and eventually death. Current diagnosis of E. scopthalmi use traditional techniques, based on the identification of the morphology of the parasite. These techniques take extended time to be carried out and do not favour the adoption of control measure at turbot farms and require the sacrifice of fish. This study develops a fast real‐time PCR molecular tool for the detection of E. scophthalmi in infected farmed turbot. This methodology is applicable for routine controls on the farm at every stage of the parasite infection. Results of the study demonstrate the robustness, specificity, efficiency and reliability of the technique. In addition, this study also provides a non‐invasive procedure of sampling through swaps. This allows control, prevention and diagnosis of the parasite infection at turbot farms while maintaining the welfare of the cultivated fish and avoiding sacrifice of the fish sampled.     

Effect of dietary supplementation of palm fruit extracts on the transcriptomes of growth,antioxidant enzyme and immune‐related genes in common carp (Cyprinus carpio) fingerlings

This study investigates the effects of date palm extracts [DPE] (Phoenix dactylifera L. Arecaceae) on growth, immune function and antioxidant system in common carp fingerlings (Cyprinus carpio). One hundred and twenty fish (4.06 ± 0.13 g) were divided into six groups fed on control diet or diets containing 200 mL kg^?1 DPE for 8 weeks. At the end of feeding trial, the expression of different genes was measured. Selected genes were grouped into three categories: growth factor genes in brain and liver [growth hormone (GH), insulin‐like growth factors I and II (IGF‐I and IGF‐II), antioxidant‐related genes in liver [glutathione S‐transferase‐alpha (GST‐α), glutathione reductase (GR) and glutathione peroxidase genes (GPX)] and immune‐related genes in head kidney [interleukin‐8 (IL‐8), interleukin‐10 (IL‐10) and transforming growth factor‐beta (TGF‐β) genes]. The relative expression of the growth‐related genes in fish fed DPE showed no significant increase compared to control group (P > 0.05). On the other hand, DPE altered the expression of genes encoding antioxidants enzymes in liver of fingerlings which was statistically significant with respect to the control samples in case GPX (P < 0.05). Also, DPE caused remarkable increases in the expression of the immune‐related genes (IL‐8, IL‐10 and TGF‐β) analysed on head kidney of common carp fingerlings compared to the control group (P < 0.05). In conclusion, it can be suggested that administration of DPE in early stages of common carp culture can promote immune efficacy and increase the antioxidant activity.     

Effects of four feeding stimulants in high plant‐based diets on feed intake,growth performance,serum biochemical parameters,digestive enzyme activities and appetite‐related genes expression of juvenile GIFT tilapia (Oreochromis sp.)

The present research evaluated the effects of four feeding stimulants on feed intake (FI), growth performance, body composition, serum biochemical parameters, digestive enzyme activities and appetite‐related gene expression for juvenile GIFT tilapia. Five experimental high plant‐based diets (with 0, 0.4 g kg^?1 dimethyl‐β‐propiothetin (DMPT), 0.6 g kg^?1 dimethylthetin (DMT), 1.8 g kg^?1 tryptophan (Trp) and 6 g kg^?1 betaine (Bet), respectively supplemented) were fed to juvenile GIFT tilapia (Oreochromis sp.) (3.34 ± 0.01 g) for 8 weeks. The results showed that FI was significantly increased by Trp and DMT. The highest specific growth rate (SGR) and protein efficiency ratio (PER) was observed for fish fed Bet diet, followed by DMT. Serum triglyceride was significantly increased by DMPT. Fish fed Bet diet showed significantly higher intestinal protease, liver protease and intestinal amylase activities. The neuropeptide Y (NPY) mRNA expression in brain was significantly improved by Trp, and ghrelin mRNA expression in stomach was significantly increased by DMT. It is concluded that supplementation of Trp or DMT could act as an effective feeding stimulants (FS) in high plant‐based diets for juvenile GIFT tilapia, which might be associated with the relatively higher orexigenic (NPY or ghrelin) genes expression, and supplementation of Bet could improve SGR, PER and reduce feed conversion ratio (FCR) of juvenile GIFT tilapia.     

The study of antioxidant enzymes and immune‐related genes expression in common carp (Cyprinus carpio) fingerlings fed different prebiotics

The aim of the present study was to study the effects of different prebiotics (galacto‐, fructooligosaccharide and inulin) on immune response and oxidative stress of common carp (Cyprinus carpio) fingerlings at the molecular level. A total number of 240 fish (13.85 ± 0.85 g) were supplied and randomly stocked in twelve fiberglass tanks (20 specimens per tank). Fish were fed a basal formulated diet (Control) or basal diet supplemented with equal level (2%) of different prebiotics (four treatments repeated in triplicated) for 8 weeks. At the end of feeding trial, the expression of immune‐related genes (interleukin 1 beta [IL‐1β], IL‐8, IL‐10, lysozyme [LYZ], tumour necrosis factor alpha [TNF‐α] and transforming growth factor beta [TGF‐β]) were determined in head kidney and intestine tissues and the expression of antioxidant‐related genes (glutathione S‐transferase [GST‐α], glutathione reductase [GR] and glutathione peroxidase genes [GPX]) were studied in intestine. The results revealed that dietary administration of prebiotics modulated the expression of immune‐related genes and the degree of expression was affected by the type of prebiotics and the organ that was used for analyses. Also, evaluation of antioxidant genes expression showed that GSTα and GR expression levels increased as a result of feeding common carp with the prebiotics. According to these findings, it can be concluded that feeding on different prebiotics had altered effects on the expression of immune and antioxidant‐related genes.     

Occurrence of multiple antibiotic resistance and genotypic characterization in Edwardsiella tarda isolated from cage‐cultured hybrid red tilapia (Oreochromis sp.) in the Ping River,Northern Thailand

Edwardsiella tarda is a pathogen that causes edwardsiellosis in aquatic animals. The emergence of multiple antibiotic‐resistant strains makes antibiotic treatment difficult. This study aimed to investigate the antibiotic susceptibility patterns and the genotypic characterization of E. tarda isolated from cage‐cultured red tilapia in Thailand. A total of 30 isolates were identified as E. tarda using biochemical and molecular analysis. The disc diffusion method for testing antibiotic susceptibility showed all the isolates were resistant to colistin sulphate and oxolinic acid. High levels of resistance to amoxicillin, ampicillin, ceftazidime, oxytetracycline and sulphamethoxazole/trimethoprim were observed as well. The multiple antibiotic resistance index ranged from 0.25 to 0.92, indicating that these isolates had been exposed to high risk sources of contamination where antibiotics were commonly used. All the isolates carried the bla_TEM gene based on polymerase chain reaction (PCR). The tetA and sul3 genes were detected in 90% (27/30) and 26.7% (8/30) of the isolates respectively. Nine different genetic groups of isolates were obtained using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC‐PCR). A correlation between genetic types and multiple antibiotic‐resistant patterns was found. These results highlight the potential risks of multiple antibiotic‐resistant isolates for humans and the environment.     

Identification of quantitative trait loci for growth‐related traits in the swimming crab Portunus trituberculatus

The swimming crab (Portunus trituberculatus) is an economically important species in Asian aquaculture. Implementing growth‐related traits of P. trituberculatus into genetic breeding programmes is an ongoing effort. We used a previously published genetic linkage map of P. trituberculatus, containing 55 simple sequence repeat (SSR) markers and 172 amplified fragment length polymorphism (AFLP) techniques to map quantitative trait loci (QTL) for growth‐related traits in a single full sibling F₂ family. Ten growth‐related traits were measured for QTL mapping. Composite interval mapping identified 9 QTL on the female map and 16 on the male map. Individual QTL with additive effects explained 11–38% of the phenotypic variance for various traits using the female parent's map, and from 1% to 21% using the male parent's map. Two QTL explaining a large percentage of variation in body weight were detected on chromosome 17 on the female map, and on chromosome 16 on the male map, and contributed 38% and 18% of the phenotypic variance respectively. This is the first study to report the detection and positioning of major QTL affecting growth in a true crab species (Brachyura). The mapping of growth‐related QTL in this study raises the possibility of improving the growth of P. trituberculatus through marker‐assisted selection.     

1H NMR‐based metabolomics studies on the effect of size‐fractionated fish protein hydrolysate,fish meal and plant protein in diet for juvenile turbot (Scophthalmus maximus L.)

This study was designed to evaluate changes in the metabolic profile of liver and muscle of turbot (Scophthalmus maximus L.) fed fishmeal‐based diet, diets containing size‐fractionated fish protein hydrolysate and plant protein‐based diet using ¹H NMR‐based metabolomics approach combined with the growth. Fish protein hydrolysate (FPH) was obtained by enzymatic treatment, permeate fraction was obtained as UF by ultrafiltered step, and retentate fraction was retained as RF. FM diet contained fish meal used as a single protein source. Four other diets (PP, UF, FPH and RF) contained 180 g kg^?1 diet fish meal. 54, 55 and 55 g kg^?1 dry diet UF, FPH and RF were supplemented to UF, FPH and RF diets. All diets were formulated to be isolipidic and isonitrogenous fed to five triplicate groups of turbot (16.05 ± 0.03 g) for 68 days. O‐PLS‐DA in FM versus UF, FM versus FPH, FM versus RF and FM versus PP resulted in a reliable model for muscle and liver tissue, while O‐PLS‐DA in UF versus FPH and UF versus RF only showed metabolites changes in liver tissue. Results indicated that metabolite changes among the different treatments were consistent with the growth tendency.     

Evaluating the most suitable nonlinear growth model for turbot (Scophthalmus maximus) in aquaculture 2 (weight application): Multi‐criteria model selection and growth prediction

Seeking the most suitable model to describe the growth of turbot, we analysed growth data of two different turbot (Scophthalmus maximus) strains reared communally in a recirculating aquaculture system. We fitted 10 different nonlinear growth models to individual weight gain data (n = 2,010) during the grow‐out phase. Analyses were carried out for each strain, for sexes within strains and for a pooled data set containing both strains and sexes. To assess the model performance, three different criteria are used. Further, a growth‐simulation was performed to evaluate the shape of the generated curve. This way we could assess the capability of the models to predict future growth. The 3‐parametric Gompertz model achieved the best fit in 42.9% of all cases tested and the lowest Bayesian information criterion in 100% of cases. The model produced realistically shaped curves and asymptotic values matching the biological attributes of the species. In contrast, 5‐parametric functions projected unrealistically shaped curves and predicted improbable mature sizes. Our results show that increasing number of parameters do not lead to increasing goodness of fit, but tend to result in overfitting, and demonstrate the advantages of the 3‐parametric Gompertz model for describing the growth of turbot.     

Genetic parameters for growth‐related traits and survival with age in the Kuruma shrimp Marsupenaeus japonicus

Genetic parameters were estimated for growth‐related traits and survival of the Kuruma shrimp Marsupenaeus japonicus based on 66 families, including 30 paternal half‐sib families, which were obtained using artificial insemination of two females by each male. The variance components for growth‐related trait and survival were estimated using a single‐trait animal model and a sir‐dam model, respectively, and genetic parameters were estimated using the restricted maximum likelihood method. The coefficients of variation for growth‐related traits such as body length (BL), abdominal length (AL), and body weight (BW) were 9.36%–22.44%, 10.61%–21.92%, and 26.47%–58.33%, respectively, at different ages (45, 75, 105, 135, and 165 days). The corresponding heritability estimates for each growth trait were 0.1545–0.1951, 0.1672–0.1905, and 0.1596–0.1934, respectively, all of which were found to be at moderate levels and increased with age. The heritability of survival on day 165 was low (0.003). The genetic correlations among growth traits were positive and high (0.7316–0.9896) at the different ages, suggesting that selection to improve any single growth trait will cause positive responses in other growth traits examined in M. japonicus. The genetic correlations between growth traits and survival were also positive but low (0.005–0.087), which indicated that only selecting for growth traits may not cause a positive correlated response in the survival of the core population. According to the above results, we suggested that growth and survival traits should be taken as improving targets of breeding in M. japonicus. These results provided reference data for selective breeding and multitrait selection of M. japonicus.