High water temperature triggers early sexual maturation in the juvenile red spotted grouper Epinephelus akaara: Via regulation of reproduction-related hormones in the brain-pituitary-gonadal axis.

The red spotted grouper Epinephelus akaara is a marine species of economic importance and also at risk of extinction. This study investigated the effects of high water temperature on the growth and maturation of juvenile E. akaara females. From 160-420 days post-hatching (dph), the fish were maintained under natural water temperature (NT) and a constant high-water temperature (HT). From 240 dph, both the total length and body weight in the HT group were greater than in NT group. After 360 dph, the gonadosomatic index was also increased in the HT group compared to NT group. Mature oocytes were only observed in the HT group at 330, 360, and 390 dph. Both kiss1 and kiss2 levels increased at 240 and 270 dph in both groups; however, they were greater in the HT group at 240 dph. Similarly, gpr54 levels after 360 dph were greater in the HT group, suggesting that kisspeptin is related to maturation via its receptor gpr54. Levels of fshß and lhß were greater in the HT group after 360 dph. Estradiol-17ß (E2) levels after 160 dph (except 300 dph) were greater in the HT group than in the NT group, suggesting that the higher E2 levels trigger maturation, and is related to increased fshß and lhß. This study provides evidence that high water temperature is effective in accelerating growth and triggering early maturation of juvenile E. akaara, via regulating gpr54, fshß, lhß, and E2 levels.

Hydrolysable tannin improves growth performance and liver health of largemouth bass (Micropterus salmoides) fed high soybean meal diets.

This study provided evidence that the inclusion of hydrolysable tannin (HT) in high soybean meal (SBM) diets improved growth performance and glycolipid metabolism of largemouth bass (Micropterus salmoides). In vivo, various levels of HT were added to high SBM diets and fed to largemouth bass (initial weight: 6.00 ± 0.03 g) for 56 days. Results showed that a high level of SBM led to the reduction in growth performance, as evidenced by decreased weight gain rate and impaired hepatic function. Dietary supplementation with HT (1.0 g/kg) improved growth performance of largemouth bass, accompanied by the enhancements in hepatic antioxidant capacity and glycolipid metabolism. In vitro, HT facilitated glucose utilization in hepatocytes and positively influenced the modulation of crucial genes within the PI3K/Akt signaling pathway. Conversely, administration of LY294002 (a PI3K inhibitor) reversed the detrimental effects observed in hepatocytes exposed to high glucose levels. Overall, incorporating HT (1.0 g/kg) into the diet enhanced liver health and improved the absorption and utilization of SBM in largemouth bass, potentially achieved through modulation of the PI3K/Akt signaling pathway.

Bentonite-supplemented diets improved fish performance ammonia excretion haemato-biochemical analyses immunity antioxidants and histological characteristics of European seabass Dicentrarchus labrax.

The purpose of this research was to examine the potential effects of bentonite (BN) supplemented diets on growth, feed utilization, blood biochemistry, and histomorphology of Dicentrarchus labrax. Six treatments in triplicate were tested: B0, B0.5, B1.0, B1.5, B3.0, and B4.5, which represented fish groups fed diets supplemented with 0, 0.5, 1, 1.5, 3, and 4.5% BN, respectively. For 84 days, juveniles' seabass (initial weight = 32.73 g) were fed diets containing 46% protein, three times daily at 3% of body weight. With a 5% daily water exchange, underground seawater (32 ppt) was used. Findings revealed significant improvements in water quality (TAN and NH3), growth (FW, WG and SGR) and feed utilization (FCR, PER and PPV) in fish fed BN-supplemented diets, with the best values in favor of the B1.5 group. Additional enhancements in kidney function indicators (urea and uric acid) and liver enzymes were observed in fish of the BN-treated groups along with a decrease in cholesterol level in the B1.5 group. Further improvements in fish innate immunity (hemoglobin, red blood cells, glucose, total protein, globulin, and immunoglobulin IgM), antioxidant activity (total antioxidative capacity and catalase), and decreased cortisol levels in fish of the BN-treated groups. Histological examinations of the anterior and posterior intestines and liver in groups B1.5 and B3 revealed the healthiest organs. This study recommends BN at a concentration of 1.5% as a feed additive in the Dicentrarchus labrax diet.

Dietary lipid sources affect growth performance, lipid deposition, antioxidant capacity and inflammatory response of largemouth bass (Micropterus salmoides).

The present study explored the effects of different lipid sources on growth performance, lipid deposition, antioxidant capacity, inflammatory response and disease resistance of largemouth bass (Micropterus salmoides). Four isonitrogenous (crude protein 50.46 %) and isolipidic (crude lipid 11.12 %) diets were formulated to contain 7 % of different oil sources including fish oil (FO) (control), soybean oil (SO), linseed oil (LO) and coconut oil (CO). Largemouth bass with initial body weight of 36.0 ± 0.2 g were randomly distributed into 12 tanks, with 30 fish per tank and 3 tanks per treatment. The fish were fed with the experiment diets twice daily for 8 weeks. The results indicated that the weight gain of largemouth bass fed the FO diet was significantly higher than that of fish fed the LO and CO diets. The liver crude lipid content in FO group was significantly higher than other groups, while the highest liver triglyceride content was showed in SO group and the lowest was detected in LO group. At transcriptional level, expression of lipogenesis related genes (pparγ, srebp1, fas, acc, dgat1 and dgat2) in the SO and CO group were significantly higher than the FO group. However, the expression of lipolysis and fatty acids oxidation related genes (pparα, cpt1, and aco) in vegetable oils groups were significantly higher than the FO group. As to the antioxidant capacity, vegetable oils significantly reduced the malondialdehyde content of largemouth bass. Total antioxidant capacity in the SO and LO groups were significantly increased compared with the FO group. Catalase in the LO group was significantly increased compared with the FO group. Furthermore, the ER stress related genes, such as grp78, atf6α, atf6ß, chop and xbp1 were significantly enhanced in the vegetable oil groups compared with the FO group. The activity of serum lysozyme in vegetable oil groups were significantly higher than in FO group. Additionally, the relative expression of non-specific immune related genes, including tlr2, mapk11, mapk13, mapk14, rela, tgf-ß1, tnfα, 5lox, il-1ß and il10, were all significantly increased in SO and CO groups compared to the other groups. In conclusion, based on the indexes including growth performance, lipid deposition, antioxidant capacity and inflammatory response, SO and LO could be alternative oil sources for largemouth bass.

Estradiol-17ß and bisphenol A affect growth and mineralization in early life stages of seabass.

Natural and synthetic estrogens are contaminants present in aquatic ecosystems. They can have significant consequences on the estrogen-sensitive functions of organisms, including skeletal development and growth of vertebrate larvae. Synthetic polyphenols represent a group of environmental xenoestrogens capable of binding the receptors for the natural hormone estradiol-17ß (E2). To better understand how (xeno-)estrogens can affect the skeleton in fish species with high ecological and commercial interest, 16 days post-hatch larvae of the seabass were experimentally exposed for 7 days to E2 and Bisphenol A (BPA), both used at the regulatory concentration of surface water quality (E2: 0.4 ng.L-1, BPA: 1.6 µg.L-1) or at a concentration 100 times higher. Skeletal mineralization levels were evaluated using Alizarin red staining, and expression of several genes playing key roles in growth, skeletogenesis and estrogen signaling pathways was assessed by qPCR. Our results show that E2 exerts an overall negative effect on skeletal mineralization at the environmental concentration of 0.4 ng.L-1, correlated with an increase in the expression of genes associated only with osteoblast bone cells. Both BPA exposures inhibited mineralization with less severe effects and modified bone homeostasis by regulating the expression of gene encoding osteoblasts and osteoclasts markers. Our results demonstrate that environmental E2 exposure inhibits larval growth and has an additional inhibitory effect on skeleton mineralization while both BPA exposures have marginal inhibitory effect on skeletal mineralization. All exposures have significant effects on transcriptional levels of genes involved in the skeletal development of seabass larvae.

A study of the potential effect of yellow mealworm (Tenebrio molitor) substitution for fish meal on growth, immune and antioxidant capacity in juvenile largemouth bass (Micropterus salmoides).

This study aimed to evaluate the effects of partial replacement of fish meal (FM) with yellow mealworm (Tenebrio molitor, TM) on the growth performance, food utilization and intestinal immune response of juvenile largemouth bass (Micropterus salmoides). Seven diets containing increasing levels of TM (FM substitution) were designed (approximately 0% (0%), 4% (11.1%), 8.1% (22.2%), 12.2% (33.3%), 16.3% (44.4%), 20.4% (55.5%), and 24.5% (66.6%), designated TM0, TM11, TM22, TM33, TM44, TM55, and TM66, respectively). 420 fish were randomly selected and placed in 21 cages (1 m*1 m*1 m, 7 treatments for triplicate, 20 fish per cage). Fish (initial weight 6.25 ± 0.03 g) were fed seven isonitrogenous (47%) and isocaloric (19 MJ kg-1) diets to satiety twice daily for 8 weeks. Compared to the control group (TM0), TM11 showed no significant difference in the weight gain rate (WGR), specific growth rate (SGR) or feed conversion ratio (FCR), while all other TM inclusion groups presented different degrees of decline. There was no significant difference in the whole-body composition among all groups (P > 0.05). Plasma total protein (TP), triglyceride (TG) and albumin (ALB) contents were significantly decreased in TM55 and TM66 (P < 0.05). The highest plasma aspartate transaminase (AST) activity was observed in TM66 (P < 0.05). TM33, TM44 and TM55 showed the lowest activities of plasma alanine amiotransferase (ALT) and alkaline phosphatase (ALP) (P < 0.05). Moreover, increased mRNA levels of superoxide dismutase (SOD) and catalase (CAT) were measured in the TM11 to TM55 groups, while intestinal SOD activity peaked in TM11 (P < 0.05). With the exception of TM11, the other TM inclusion groups showed significant inhibition of the relative expression of RelA, C3 and TNF-α (P < 0.05). All experimental groups exhibited lower expression of IL-10 than TM0 (P < 0.05). The TM11 group showed significantly upregulated expression of IL-1ß and TGF-ß (P < 0.05). In addition, TLR2 expression was increased in TM11 and TM22 (P < 0.05). Considering enzyme activities and immune-related gene expression, TM supplementation levels should not exceed 4% (TM11).

The influence of 17ß-oestradiol on lymphopoiesis and immune system ontogenesis in juvenile sea bass, Dicentrarchus labrax.

The female sex steroid 17ß-oestradiol (E2) is involved in the regulation of numerous physiological functions, including the immune system development and performance. The role of oestrogens during ontogenesis is, however, not well studied. In rodents and fish, thymus maturation appears to be oestrogen-dependent. Nevertheless, little is known about the function of oestrogen in immune system development. To further the understanding of the role of oestrogens in fish immune system ontogenesis, fingerlings of European sea bass (Dicentrarchus labrax) were exposed for 30 days to 20 ng E2·L-1, at two ages tightly related to thymic maturation, i.e., 60 or 90 days post hatch (dph). The expression of nuclear and membrane oestrogen receptors was measured in the thymus and spleen, and the expression of several T cell-related gene markers was studied in both immune organs, as well as in the liver. Waterborne E2-exposure at 20.2 ± 2.1 (S.E.) ng·L-1 was confirmed by radioimmunoassay, leading to significantly higher E2-contents in the liver of exposed fish. The majority of gene markers presented age-dependent dynamics in at least one of the organs, confirming thymus maturation, but also suggesting a critical ontogenetic window for the implementation of liver resident γδ and αß T cells. The oestrogen receptors, however, remained unchanged over the age and treatment comparisons with the exception of esr2b, which was modulated by E2 in the younger cohort and increased its expression with age in the thymus of the older cohort, as did the membrane oestrogen receptor gpera. These results confirm that oestrogen-signalling is involved in thymus maturation in European sea bass, as it is in mammals. This suggests that esr2b and gpera play key roles during thymus ontogenesis, particularly during medulla maturation. In contrast, the spleen expressed low or non-detectable levels of oestrogen receptors. The E2-exposure decreased the expression of tcrγ in the liver in the cohort exposed from 93 to 122 dph, but not the expression of any other immune-related gene analysed. These results indicate that the proliferation/migration of these innate-like T cell populations is oestrogen-sensitive. In regard to the apparent prominent role of oestrogen-signalling in the late thymus maturation stage, the thymic differentiation of the corresponding subpopulations of T cells might be regulated by oestrogen. To the best of our knowledge, this is the first study investigating the dynamics of both nuclear and membrane oestrogen receptors in specific immune organs in a teleost fish at very early stages of immune system development as well as to examine thymic function in sea bass after an exposure to E2 during ontogenesis.

Dietary valine levels affect growth, protein utilisation, immunity and antioxidant status in juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

A 6-week growth trial was conducted to evaluate the influences of dietary valine (Val) levels on growth, protein utilisation, immunity, antioxidant status and gut micromorphology of juvenile hybrid groupers. Seven isoenergetic, isoproteic and isolipidic diets were formulated to contain graded Val levels (1·21, 1·32, 1·45, 1·58, 1·69, 1·82 and 1·94 %, DM basis). Each experimental diet was hand-fed to triplicate groups of twelve hybrid grouper juveniles. Results showed that weight gain percentage (WG%), protein productive value, protein efficiency ratio, and feed efficiency were increased as dietary Val level increased, reaching a peak value at 1·58 % dietary Val. The quadratic regression analysis of WG% against dietary Val levels indicated that the optimum dietary Val requirement for hybrid groupers was estimated to be 1·56 %. Gut micromorphology and expression of growth hormone in pituitary, insulin-like growth factor 1, target of rapamycin and S6 kinase 1 in liver were significantly affected by dietary Val levels. In serum, fish fed 1·58 % dietary Val had higher superoxide dismutase, catalase, lysozyme activities and IgM concentrations than fish fed other dietary Val levels. Fish fed 1·58 % dietary Val had higher expression of NF-E2-related factor 2 in head kidney than fish fed other dietary Val levels. Generally, the optimum dietary Val requirement for maximal growth of hybrid groupers was estimated to be 1·56 % of DM, corresponding to 3·16 % of dietary protein, and dietary Val levels affected growth, protein utilisation, immunity and antioxidant status in hybrid groupers.

Total replacement of fishmeal with poultry by-product meal affected the growth, muscle quality, histological structure, antioxidant capacity and immune response of juvenile barramundi, Lates calcarifer.

The present study investigates if the total replacement of dietary fishmeal (FM) with poultry by-product meal (PBM), supplemented with methionine influences the muscle fatty acids composition, normal gut morphology, histological traits of the liver, muscle, and gill, liver enzymes, immune and antioxidant response, and stress-related gene in juvenile barramundi, Lates calcarifer in relation to growth and feed utilization. Barramundi (3.58±0.01g) were randomly distributed into six 300 L seawater recirculating tanks (25 fish/tank) and fed two formulated isonitrogenous and isolipidic diets for 6 weeks. The control diet had FM as the sole animal protein source, whereas other test diet had only PBM as an animal protein source. Dietary PBM affected the fish performance and feed utilization. Regarding muscle fatty acid profile, total saturated fatty acids and monounsaturated fatty acids elevated while total PUFA particularly n-3 LC-PUFA and EPA decreased in PBM fed fish than control diet fed fish. Liver, muscle, gill, and intestinal histology showed no obvious alteration in control diet fed fish, however, more lipid droplets and hepatic vacuolization in the liver, necrotic myotome in muscle, hyperplasia in secondary lamellae in gill and short and broken folds in the intestine were observed in PBM fed fish. Similar to light microscopy observation of intestinal morphology, the transmission electron microscopy (TEM) analysis revealed shorter and smaller microvilli in fish fed PBM. Histopathological alterations in the liver of PBM fed fish were further associated with the elevated levels of aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH) and the significant upregulation of stress-related genes, HSP70 and HSP90. Also, a negative influence on lysozyme activity, and antioxidant enzymatic activities were recorded in fish fed PBM. Overall, it can be concluded that a total substitution of FM protein by methionine supplemented PBM negatively influenced the growth performance, liver health, histological traits of different organs, immune and antioxidant response, and expression of stress-related genes in juvenile barramundi.

Dietary vitamin A deficiency reduces growth performance, immune function of intestine, and alters tight junction proteins of intestine for juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

This study was carried out to investigate the effects of dietary vitamin A (VA) on growth performance, antioxidant capacity, digestion, intestinal immune response, and mRNA expression of intestinal tight junction proteins for juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Six isonitrogenous and isolipidic experimental diets were formulated to obtain VA levels (317, 1136, 2038, 4142, 7715, 15204 IU/kg diet, respectively). The triplicate groups of fish (average weight of 9.01 ± 0.27 g) were fed twice daily (8:00 and 16:00) for 7 weeks. Based on the broken-line analysis model of WG and LYZ activity, the dietary VA requirement of hybrid grouper were estimated to be 2688.58 and 4096.36 IU/kg diet. The results showed that VA deficiency or excess could reduce Weight gain, specific growth rate, and protein efficiency ratio, and increase feed conversion ratio and hepatosomatic index (P < 0.05). In addition, VA deficiency could reduce the serum activities of acid phosphatase (ACP), superoxide dismutase, and total antioxidant capacity and increase the malondialdehyde content (P < 0.05). VA deficiency also could reduce intestinal activities of ACP, alkaline phosphatase, lysozyme, complement 3, complement 4 contents, and activities of alpha-amylase, lipase, and trypsin (P < 0.05). Meanwhile, VA deficiency could reduce villus height in proximal intestine (PI) and mid intestine (MI), as well as muscle thickness in PI and distal intestine (DI) (P < 0.05). Moreover, VA deficiency could down-regulated antimicrobial peptides (ß-defensin, Hepcidin [not in MI and DI], Epinecidin), anti-inflammatory cytokines (interleukin 10 and transforming growth factor ß1 [not in DI]), tight junction proteins (occluding and claudin3) mRNA levels in the PI, MI and DI, and up-regulated pro-inflammatory cytokines (tumor necrosis factor α [not in MI] and interleukin 1ß [not in MI]), signaling molecules c-Rel and p65 (P < 0.05). Collectively, VA deficiency could reduce growth performance because of a negative effect on intestinal health by depressing digestive abilities, intestinal morphology, immunity and tight junction function in the intestine.

Assessment of dietary supplementation with galactomannan oligosaccharides and phytogenics on gut microbiota of European sea bass (Dicentrarchus Labrax) fed low fishmeal and fish oil based diet.

There is an increasing interest from the aquafeed industry in functional feeds containing selected additives that improve fish growth performance and health status. Functional feed additives include probiotics, prebiotics, organic acids, and phytogenics (substances derived from plants and their extracts). This study evaluated the effects of dietary inclusion of a mucilage extract rich in galactomannan oligosaccharides (GMOS), a mixture of garlic and labiatae-plants oils (PHYTO), and a combination of them (GMOSPHYTO), on gut microbiota composition of European sea bass (Dicentrarchus labrax) fed with a low fishmeal (FM) and fish oil (FO) diet. Three experimental diets and a control diet (plant-based formulation with 10% FM and 6% FO) were tested in a 63-days feeding trial. To analyze the microbiota associated to feeds and the intestinal autochthonous (mucosa-adhered) and allochthonous (transient) microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and QIIME2 pipeline were used. Metabarcoding analysis of feed-associated bacteria showed that the microbial communities of control (CTRL) feed deeply differed from those of experimental diets. The number of reads was significantly lower in CTRL feed than in other feeds. The OTU (operational taxonomic unit) number was instead similar between the feeds, ranging from 42 to 50 OTUs. The variation of resident gut microbiota induced by diet was lower than the variation of transient intestinal microbiota, because feedstuffs are a major source of allochthonous bacteria, which can temporarily integrate into the gut transient microbiome. However, the composition of transient bacterial communities was not simply a mirror of feed-borne bacteria. Indeed, the microbial profile of feeds was different from both faecal and mucosa profiles. Our findings suggest that the dietary inclusion of GMOS (0.5%) and PHYTO (0.02%) in a low FM and FO diet induces changes in gut microbiota composition of European sea bass. However, if on allochthonous microbiota the combined inclusion of GMOS and PHYTO showed an antagonistic effect on bactericidal activity against Vibrionales, at mucosa level, only GMOSPHYTO diet increased the relative abundance of Bacteroidales, Lactobacillales, and Clostridiales resident bacterial orders. The main beneficial effects of GMOS and PHYTO on gut microbiota are the reduction of coliforms and Vibrionales bacteria, which include several potentially pathogenic species for fish, and the enrichment of gut microbiota composition with butyrate producer taxa. Therefore, these functional ingredients have a great potential to be used as health-promoting agents in the farming of European sea bass and other marine fish.

Inter-individual variability in freshwater tolerance is related to transcript level differences in gill and posterior kidney of European sea bass.

Acclimation to low salinities is a vital physiological challenge for euryhaline fish as the European sea bass Dicentrarchus labrax. This species undertakes seasonal migrations towards lagoons and estuaries where a wide range of salinity variations occur along the year. We have previously reported intraspecific differences in freshwater tolerance, with an average 30% mortality rate. In this study, we bring new evidence of mechanisms underlying freshwater tolerance in sea bass at gill and kidney levels. In fresh water (FW), intraspecific differences in mRNA expression levels of several ion transporters and prolactin receptors were measured. We showed that the branchial Cl-/HCO3- anion transporter (slc26a6c) was over-expressed in freshwater intolerant fish, probably as a compensatory response to low blood chloride levels and potential metabolic alkalosis. Moreover, prolactin receptor a (prlra) and Na+/Cl- cotransporter (ncc1) but not ncc-2a expression seemed to be slightly increased and highly variable between individuals in freshwater intolerant fish. In the posterior kidney, freshwater intolerant fish exhibited differential expression levels of slc26 anion transporters and Na+/K+/2Cl- cotransporter 1b (nkcc1b). Lower expression levels of prolactin receptors (prlra, prlrb) were measured in posterior kidney which probably contributes to the failure in ion reuptake at the kidney level. Freshwater intolerance seems to be a consequence of renal failure of ion reabsorption, which is not sufficiently compensated at the branchial level.

Gene networks and toxicity/detoxification pathways in juvenile largemouth bass (Micropterus salmoides) liver induced by acute lead stress.

Lead (Pb) is a nonessential heavy metal that can be bioconcentrated to highly toxic levels in the environment. To understand the genes and toxicity/detoxification pathways of juvenile largemouth bass, liver transcriptomes were investigated in this fish after acute 96 h Pb exposure (Pb nitrate 0, 17.8, or 100 mg/L). Acute Pb exposure induced an immune response and apoptosis pathway activation in the liver. A number of transcripts related to complement and coagulation cascades were significantly increased. Up- and downregulated genes were significantly enriched in numerous pathways, including the natural killer cell-mediated cytotoxicity pathway, the Jak-STAT and P53 signaling pathways, cancer and apoptosis. These genes included Bid，Bcl-2, JNK, and PI3K (17.8 mg/L) and PI3K, AKT, PPARδ, RAS, MMPs, c-Jun p53, and PD-L1 (100 mg/L). Comprehensive analysis of liver transcriptomic data revealed numerous pathways associated with the immune system and carcinogenesis, especially pathways related to apoptosis and systemic lupus erythematosus.

Effects of dietary n-3 highly unsaturated fatty acids on growth, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu).

This study was conducted to investigate the effects of dietary n-3 highly unsaturated fatty acids (n-3 HUFA) on growth performance, non-specific immunity, expression of some immune-related genes and resistance to Vibrio harveyi in juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ Epinephelus lanceolatu). Six isoproteic and isolipidic experimental diets were formulated with graded levels of n-3 HUFA (0.65, 1.00, 1.35, 1.70, 2.05 and 2.40% of dry matter, respectively), and the 0.65% group was used as control group. Each diet was randomly allocated to triplicate groups of fish in 1000 L fiberglass tank, and each tank was stocked with 40 fish (initial weight 12.06 ± 0.01 g) for 8 weeks. Results showed that feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI) and condition factor (CF) were all not significantly affected by dietary n-3 HUFA levels (P > 0.05). Weight gain (WG) and specific growth rate (SGR) in 1.35% group were significantly higher than those in 2.40% group (P < 0.05). Crude lipid of body in 1.00% group was significantly lower than that in 1.70% and 2.40% groups (P < 0.05). Liver and muscle fatty acid profiles reflected that of diets. Before challenge with Vibrio harveyi, the activity of serum superoxide dismutase (SOD), catalase (CAT) and content of complement 3 (C3) in 1.35% and 1.70% groups significantly higher than those of control group (P < 0.05). After challenge with Vibrio harveyi, serum CAT, glutathione peroxidase (GSH-PX), lysozyme (LZM) and C3 all increased sharply, while SOD showed the opposite trend. Before challenge with Vibrio harveyi, the expression levels of intestine toll-like receptor 22 (TLR22) and myeloid differentiation factor 88 (MyD88) mRNA in 2.40% group were significantly increased, and the expression levels of tumour necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) mRNA in 2.05% group were significantly higher than those in 1.00% and 1.35% groups (P < 0.05). In addition, the TLR22 and IL-1ß mRNA levels in kidney of 1.70% group were significantly lower than those in control group (P < 0.05). After challenge with Vibrio harveyi, the expression level of MyD88 mRNA in intestine of 1.35% group was significantly higher than that in 1.00% group and from 1.70% to 2.40% groups (P < 0.05), while TNF-α and IL-1ß obtained minimum values in 1.70% group. In the kidney, the interleukin 10 (IL10) mRNA expression was significantly higher in 1.70% group than that in other groups, while the IL-1ß expression in 1.70% group was on the contrary and significantly lower than that in 2.40% group (P < 0.05). Results of this study suggested that moderate dietary n-3 HUFA (1.47%-1.70% HUFA) could improve the growth performance, non-specific immunity and inhibit the inflammatory response of hybrid grouper.

Efficacy of purified nucleotide supplements on the growth performance and immunity of hybrid striped bass Morone chrysops x Morone saxatilis.

Fishmeal is being increasingly replaced in aquatic animal diets with alternative plant protein feedstuffs such as soybean meal which have lower concentrations of nucleotides; therefore, supplemental sources of exogenous nucleotides in diets could become increasingly important. A 9-week feeding trial was conducted with triplicate groups of juvenile hybrid striped bass (average initial body weight ± standard deviation, 5.6 ± 0.1 g) to determine the effects of supplementing single purified nucleotides on the growth performance and immune parameters. The basal diet, which utilized menhaden fishmeal (25%) and soybean meal (75%) as protein sources, contained 44% protein, 10% lipid and an estimated digestible energy level of 3.5 kcal g-1. Single additions of 5'- adenosine monophosphate (AMP), 5'- uridine monophosphate (UMP), 5'- cytidine monophosphate (CMP), 5'- guanosine monophosphate (GMP), and 5'- inosine monophosphate (IMP) disodium salts (Chem-Impex International, Wood Dale, Illinois, USA) were evaluated with each nucleotide added to the basal diet at 0.5% of dry weight at the expense of cellulose. A positive control diet in this trial was a diet containing 5'- AMP from Sigma-Aldrich also supplemented at 0.5% by weight. Results showed significantly (P < 0.05) improved weight gain between fish fed AMP-supplemented diets and the basal diet. No statistical significance (P > 0.05) was detected in whole-body proximate composition and protein retention of fish fed any of the dietary treatments. The respiratory burst of whole blood phagocytes also was significantly (P < 0.05) higher in fish fed the AMP Sigma diet compared to the other dietary treatments. Dietary IMP and AMP both significantly (P < 0.05) enhanced the capacity of isolated phagocytes to generate extracellular superoxide anion compared to all other dietary treatments. No significant differences were seen in other innate immune parameters such as plasma lysozyme, total plasma protein, and total immunoglobulin. The ability of isolated B lymphocytes to proliferate prompted by the presence of lipopolysaccharides was significantly (P < 0.05) different among dietary treatments with the highest simulation index observed in fish fed the diets containing AMP Sigma and UMP; however, it was not significantly different from that of fish fed the basal diet. Based on all the measured responses, it is concluded that AMP at 0.5% of diet had the most positive influence on growth performance and innate immunostimulation of hybrid striped bass.

Effects of dietary yeast hydrolysate on the growth, antioxidant response, immune response and disease resistance of largemouth bass (Micropterus salmoides).

A 56-day growth trial was conducted to investigate the effects of dietary yeast hydrolysate on the growth performance, antioxidation, immune response and resistance against Aeromonas hydrophila in largemouth bass. Four experimental diets were prepared with yeast hydrolysate levels of 0% (Y0), 1.5% (Y1.5), 3.0% (Y3.0) and 4.5% (Y4.5). Each diet was randomly assigned to triplicate 150-L tanks and each tank was stocked with 30 largemouth bass (initial body weight, IBW = 7.71 ±â€¯0.02 g). A challenge test was carried out after the feeding trial by injecting A. hydrophila intraperitoneally for 4-day observation. The results showed that the FBW and WGR in Y1.5 group were significantly higher than those in Y0 group (P < 0.05) and the feed conversion ratio (FCR) got the lowest value in Y1.5 group. And the hydrolysate supplement significantly increased the 4-day cumulative survival rate after the bacterial challenge (P < 0.05). The plasma malondialdehyde was lower in the yeast hydrolysate supplement groups in both pre- and post-challenge test (P < 0.05), while the plasma C3 increased (P < 0.05). In post-challenge test, the plasma superoxide dismutase (SOD) and catalase (CAT) activities increased in the Y1.5 and Y3.0 groups respectively (P < 0.05), and plasma lysozyme in Y1.5 group and the plasma IgM in Y3.0 group were higher than those in others respectively (P < 0.05). For the q-PCR results, in post-challenge test, the hepatic hep2 expression level in Y1.5 and Y4.5 groups were both significantly higher than those in others (P < 0.05), as well as il-8 in Y3.0 group. The spleen hif-1alpha and tgf-beta1 expression levels in Y4.5 group were all significantly lower than those in others (P < 0.05), while the gilt was significantly higher (P < 0.05) in the post-challenge test. And the expression levels of spleen tnf-alpah1 in Y1.5 and Y3.0 groups and il-8 in Y3.0 group were all significantly higher than those in other groups (P < 0.05) in the post-challenge test. The head kidney gilt expression level was significantly higher in the yeast hydrolysate supplement groups compared with the Y0 group (P < 0.05), and the head kidney il-8 expression level in Y1.5 group was significant higher than those in other groups in post-challenge test (P < 0.05). The present results indicated dietary yeast hydrolysate improved the antioxidant ability and enhanced the immune response of largemouth bass without negative effect on growth. And 1.5% or 3.0% of dietary yeast hydrolysate was recommended for largemouth bass based on the present results.

Dietary phytogenics and galactomannan oligosaccharides in low fish meal and fish oil-based diets for European sea bass (Dicentrarchus labrax) juveniles: Effects on gut health and implications on in vivo gut bacterial translocation.

European sea bass were fed four low FM/FO (10%/6%) diets containing galactomannan oligosaccharides (GMOS), a mixture of garlic oil and labiatae plants oils (PHYTO), or a combination of both functional products (GMOSPHYTO) for 63 days before exposing the fish to an intestinal Vibrio anguillarum infection combined with crowding stress. In order to evaluate functional diets efficacy in terms of gut health maintenance, structural, cellular, and immune intestinal status were evaluated by optical and electron microscopy and gene expression analyses. A semi-automated software was adapted to determine variations in goblet cell area and mucosal mucus coverage during the challenge test. Feeding with functional diets did not affect growth performance; however, PHYTO and GMOS dietary inclusion reduced European sea bass susceptibility to V. anguillarum after 7 days of challenge testing. Rectum (post-ileorectal valve) showed longer (p = 0.001) folds than posterior gut (pre-ileorectal valve), whereas posterior gut had thicker submucosa (p = 0.001) and higher mucus coverage as a result of an increased cell density than rectum. Functional diets did not affect mucosal fold length or the grade of granulocytes and lymphocytes infiltration in either intestinal segment. However, the posterior gut fold area covered by goblet cells was smaller in fish fed GMOS (F = 14.53; p = 0.001) and PHYTO (F = 5.52; p = 0.019) than for the other diets. PHYTO (F = 3.95; p = 0.049) reduced posterior gut goblet cell size and increased rodlet cell density (F = 3.604; p = 0.068). Dietary GMOS reduced submucosal thickness (F = 51.31; p = 0.001) and increased rodlet cell density (F = 3.604; p = 0.068) in rectum. Structural TEM analyses revealed a normal intestinal morphological pattern, but the use of GMOS increased rectum microvilli length, whereas the use of PHYTO increased (p≤0.10) Ocln, N-Cad and Cad-17 posterior gut gene expression. After bacterial intestinal inoculation, posterior gut of fish fed PHYTO responded in a more controlled and belated way in terms of goblet cell size and mucus coverage in comparison to other treatments. For rectum, the pattern of response was similar for all dietary treatments, however fish fed GMOS maintained goblet cell size along the challenge test.

Characterization of the European Sea Bass (Dicentrarchus labrax) Gonadal Transcriptome During Sexual Development.

The European sea bass is one of the most important cultured fish in Europe and has a marked sexual growth dimorphism in favor of females. It is a gonochoristic species with polygenic sex determination, where a combination between still undifferentiated genetic factors and environmental temperature determines sex ratios. The molecular mechanisms responsible for gonadal sex differentiation are still unknown. Here, we sampled fish during the gonadal developmental period (110 to 350 days post fertilization, dpf), and performed a comprehensive transcriptomic study by using a species-specific microarray. This analysis uncovered sex-specific gonadal transcriptomic profiles at each stage of development, identifying larger number of differentially expressed genes in ovaries when compared to testis. The expression patterns of 54 reproduction-related genes were analyzed. We found that hsd17ß10 is a reliable marker of early ovarian differentiation. Further, three genes, pdgfb, snx1, and nfy, not previously related to fish sex differentiation, were tightly associated with testis development in the sea bass. Regarding signaling pathways, lysine degradation, bladder cancer, and NOD-like receptor signaling were enriched for ovarian development while eight pathways including basal transcription factors and steroid biosynthesis were enriched for testis development. Analysis of the transcription factor abundance showed an earlier increase in females than in males. Our results show that, although many players in the sex differentiation pathways are conserved among species, there are peculiarities in gene expression worth exploring. The genes identified in this study illustrate the diversity of players involved in fish sex differentiation and can become potential biomarkers for the management of sex ratios in the European sea bass and perhaps other cultured species.

Advanced puberty triggered by bi-weekly changes in reproductive factors during the photolabile period in a male teleost fish, Dicentrarchus labrax L.

This study evaluated the impact of continuous light (LL) within the photolabile period on advanced puberty in juvenile male European sea bass. The exposure to an LL regime for 1 month, from August 15 to September 15 (LLa/s), was compared to a constant simulated natural photoperiod (NP) and constant continuous light conditions year-round (LLy). Somatic growth, hormone plasma levels, rates of testicular maturation and spermiation, as well as the mRNA levels of some reproductive genes were analyzed. Our results demonstrated that both LLa/s and LLy treatments, which include LL exposure during the photolabile period, were highly effective in inhibiting the gametogenesis process that affects testicular development, and clearly reduced the early sexual maturation of males. Exposure to an LL photoperiod affected body weight and length of juvenile fish during early gametogenesis and throughout the first year of life. Interestingly, LL induced bi-weekly changes in some reproductive factors affecting Gnrh1 and Gnrh2 content in the brain, and also reduced pituitary fshß expression and plasmatic levels of 11-KT, E2, Fsh throughout early gametogenesis. We suggest that low levels of E2 in early September in the LL groups, which would be concomitant with the reduced number of spermatogonial mitoses in these groups, might indicate a putative role for estrogens in spermatogonial proliferation during the early gonadal development of this species. Furthermore, a significant decrease in amh expression was observed, coinciding with low plasma levels of 11-KT under LL regimes, which is consistent with the idea that this growth factor may be crucial for the progress of spermatogenesis in male sea bass.

Assessing the long-term effect of exposure to dispersant-treated oil on fish health using hypoxia tolerance and temperature susceptibility as ecologically relevant biomarkers.

The ecological and economic importance of fish act as a brake on the development of chemical dispersants as operational instruments following oil spills. Although a valuable and consistent body of knowledge exists, its use in spill response is limited. The objective of the present study was to increase current knowledge base to facilitate the translation of published data into information of operational value. Thus we investigated the dose-response relationship between dispersant-treated oil exposure and ecologically relevant consequences by combining laboratory and field experiments. Effects were examined over almost a year using juveniles of the slowly growing, commercially important European sea bass (Dicentrarchus labrax). A reliable interpretation of biomarker responses requires a complete knowledge of the factors likely to affect them. Interpopulational variability is of particular importance in environmental impact assessment because biomarker responses from a population collected in an impacted area are classically compared with those collected in a clean site. Our study revealed no effect of the exposure to dispersant-treated oil on fish hypoxia tolerance and temperature susceptibility at 1 and 11 mo post exposure. Similarly, no effect of the exposure was observed on the ability of the fish to cope with environmental contingencies in the field, regardless of the dose tested. Thus we feel confident to suggest that a 48-h exposure to chemically treated oil does not affect the ability of sea bass to cope with mild environmental contingencies. Finally, investigation of interpopulation variability revealed large differences in both hypoxia tolerance and temperature susceptibility among the 2 populations tested, suggesting that this variability may blur the interpretation of population comparisons as classically practiced in impact assessment. Environ Toxicol Chem 2019;38:210-221. © 2018 SETAC.