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Astaxanthin (AST), functioning as an efficient antioxidant and pigment, is one of the most expensive additives in shrimp feeds. How to improve the uptake efficiency of dietary astaxanthin into farmed shrimp is of significance. The present study investigated the effects of lysophosphatidylcholine (LPC), an emulsifier, on dietary astaxanthin efficiency, growth performance, body color, body composition, as well as lipid metabolism of juvenile Pacific white shrimp (average initial body weight: 2.4 g). Three diets were prepared: control group, the AST group (supplemented with 0.02% AST), and the AST + LPC group (supplemented with 0.02% AST and 0.1% LPC). Each diet was fed to triplicate tanks, and each tank was stocked with 30 shrimp. The shrimp were fed four times daily for eight weeks. The AST supplementation improved the growth of white shrimp, while LPC further promoted the final weight of shrimp, but the whole-shrimp proximate composition and fatty acid composition were only slightly affected by AST and LPC. The LPC supplementation significantly increased the astaxanthin deposition in the muscle. The LPC supplementation significantly increased the shell yellowness of both raw and cooked shrimp compared to the AST group. Moreover, the dietary LPC increased the high-density lipoprotein-cholesterol content but decreased the low-density lipoprotein-cholesterol content in the serum, indicating the possible regulation of lipid and cholesterol transport. The addition of astaxanthin significantly up-regulated the expression of npc2 in the hepatopancreas compared to the control group, while the addition of LPC down-regulated the expression of mttp compared to the AST group. In conclusion, the LPC supplementation could facilitate the deposition of dietary astaxanthin into farmed shrimp and further enlarge the beneficial effects of dietary astaxanthin. LPC may also independently regulate shrimp body color and cholesterol transportation. This was the first investigation of the promoting effects of LPC on dietary astaxanthin efficiency. [ABSTRACT FROM AUTHOR]

Simple Summary: With the rapid development of aquaculture, searching for alternative lipid sources to fish oil has become an important task for the aqua-feed industry. Lard is regarded as a valuable alternative lipid source. Basa fish (Pangasius bocourti) offal oil also has a large annual production and high potential to be used in fish feeds. Evaluation of the efficacy of lard and Basa fish offal oil in fish feeds will provide useful information for their application in aquaculture. Both lard and Basa fish offal oil have high levels of saturated fatty acids and monounsaturated fatty acids. The "n-3 LC-PUFA sparing effects" of saturated fatty acids and monounsaturated fatty acids in fish feeds have been observed in many fish species. The present study was also aimed at validating the "n-3 LC-PUFA sparing effects" of saturated fatty acids and monounsaturated fatty acids in lard and Basa fish offal oil. Moreover, the effects of dietary lard and Basa fish offal oil on fish body composition and muscle quality were also evaluated. Lard (LD) and Basa fish offal oil (BFO) have similar fatty acid profiles, both containing high contents of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA). The present study aimed to investigate the efficacy of partial or complete replacement of marine fish oil (MFO, herring oil) by LD or BFO in the diets of tiger puffer. The control diet contained 49.1% crude protein and 9.28% crude lipid content including 6% added MFO. In other diets, 1/3, 2/3, and 3/3 of the added MFO was replaced by LD or BFO, respectively. Each diet was fed to triplicate tanks of juvenile fish (initial body weight, 13.88 g). A 46-day feeding trial was conducted in a flow-through seawater system. Each diet was fed to triplicate 200-L rectangular polyethylene tanks, each of which was stocked with 30 fish. Fish were fed to satiation three times a day. The complete replacement of added MFO (replacing 65% of the total crude lipid) had no adverse effects on fish growth performance in terms of survival (>94%), weight gain (360–398%), feed intake (2.37–3.04%), feed conversion ratio (0.84–1.02), and somatic indices. The dietary LD or BFO supplementation also had marginal effects on fish body proximate composition, biochemical parameters, muscle texture, and water-holding ability, as well as the hepatic expression of lipid metabolism-related genes. Partial (2/3) replacement of added MFO by LD or BFO did not significantly reduce the muscle n-3 LC-PUFA content, indicating the n-3 LC-PUFA sparing effects of SFA and MUFA in LD and BFO. In general, dietary LD or BFO reduced the peroxidation level and led to significant changes in the muscle volatile flavor compound profile, which were probably attributed to the change in fatty acid composition. The results of this study evidenced that LD and BFO are good potential lipid sources for tiger puffer feeds. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the effects of combined replacement of fishmeal (FM) and fish oil (FO) with poultry byproduct meal (PBM) and mixed oil (MO, poultry oil: coconut oil = 1 : 1) on growth performance, body composition and muscle quality of tiger puffer (Takifugu rubripes). Fish with an average initial body weight of 14.29 g were selected for the feeding experiment. FM accounting for 0%, 5%, and 10% of the diet was replaced by PBM. For each grade of FM replacement, 5% FO or MO was used as added oil. The six experimental diets were designated as FO-FM, MO-FM, FO-5PBM, MO-5PBM, FO-10PBM, and MO-10PBM, respectively. Each treatment was performed in triplicate with 30 fish per replicate. The feeding period was 45 days. There was no significant difference in growth performance among the groups. Dietary supplementation of both PBM and MO had marginal effects on whole-fish proximate composition, except that dietary MO supplementation significantly increased the liver moisture content. In serum, there were no significant differences in contents of triglyceride, total cholesterol, total bile acid, and protein carbonyl among groups, but the malondialdehyde content was reduced by MO. The fatty acid composition in fish mirrored those in the diets, but the omega-3 sparing effects of saturated and monounsaturated fatty acid in MO can still be observed. Dietary PBM and MO had marginal effects on free amino acid composition and texture of fish muscle, but exerted complicated effects on the muscle volatile flavor compound composition. In conclusion, combined fishmeal (10% of the diet) and fish oil (5% of the diet) replacement with poultry byproduct and mixed oil (poultry oil + coconut oil) had no adverse effects on the growth performance and body proximate composition of farmed tiger puffer. However, these replacements changed the muscle flavor compound profile. [ABSTRACT FROM AUTHOR]

This study characterized four cholesterol transport-related genes, namely, lcat, acat1, acat2, and mttp, in juvenile tiger puffer in terms of tissue distribution (eye, heart, brain, skin, liver, spleen, muscle, and intestine) and nutritional regulation. Three feeding trials were conducted: (i) using diets with different cholesterol levels (0.11%, 0.65%, 1.10%, 2.32%, and 4.59% of dry matter); (ii) using diets with different lipid levels (8.05%, 12.02%, and 16.36% of dry matter); and (iii) a 1-month starvation experiment with different sampling times (Days 1, 4, 9, 16, and 31). The lcat, acat1, and acat2 were the most abundantly expressed in the liver, while mttp was the most abundantly expressed in the intestine. The lcat had a medium expression level in the muscle and skin but the lowest expression level in the spleen and intestine. The acat1 and acat2 had similar tissue distribution patterns, except that acat2 had a lower expression level in the heart but a higher level in the intestine than acat1. The mttp had medium expression levels in the liver and spleen but had the lowest level in the eye and skin. Different dietary cholesterol levels did not significantly affect the expression of these cholesterol transport genes in the liver and intestine, except that higher cholesterol levels (1.10%–4.59%) significantly downregulated the acat2 expression in the liver. Dietary lipid levels also had a mild influence on the hepatic expression of these genes, except that the diet with 12.02% lipid resulted in significantly higher acat1 expression than the one with 16.36% lipid. The starvation time significantly affected the hepatic expression of these genes. Long-term starvation (16 and 31 days) generally upregulated the hepatic gene expression of lcat, acat1, and mttp but generally downregulated that of acat2. This study provided preliminary knowledge about the tissue distribution and nutritional regulation of cholesterol transport-related genes in marine teleost. [ABSTRACT FROM AUTHOR]

Acute hypoxia is a common stress in aquaculture, and causes energy deficiency, oxidative damage and death in fish. Many studies have confirmed that acute hypoxia activated hif1α expression, anaerobic glycolysis and antioxidant system in fish, but the effects of acute hypoxia on lipid and protein metabolism, organelle damage, and the functions of hif2α and hif3α in economic fishes have not been well evaluated. In the present study, turbot was exposed to acute hypoxia (2.0 ± 0.5 mg/L) for 6 h, 12 h, and 24 h, respectively. Then, the contents of hemoglobin (HB), metabolite, gene expressions of hifα isoforms, energy homeostasis, endoplasmic reticulum (ER) stress, and apoptosis were measured. The results suggested that turbot is intolerant to acute hypoxia and the asphyxiation point is about 1.5 mg/L. Acute hypoxia induced perk-mediated ER stress, and increased lipid peroxidation and liver injury in turbot. The blood HB level and liver vegfab expression were increased under hypoxia, which enhances oxygen transport. At hypoxia stress, hif3α, anaerobic glycolysis-related genes expression, and lactate content were increased in the liver, and glycogen was broken down to ensure ATP supply. Meanwhile, hif2α, lipid synthesis-related genes expression, and TG content were increased in the liver, but the lipid catabolism and protein synthesis were suppressed during hypoxia, which reduced the oxygen consumption and ROS generation. Our results systematically illustrate the metabolic and physiological changes under acute hypoxia in turbot, and provide important guidance to improve hypoxia tolerance in fish. [ABSTRACT FROM AUTHOR]

Simple Summary: Enteromorpha prolifera is a type of green marine macroalgae that can cause green tides due to eutrophication. However, aquafeeds demand for E. prolifera meal has gradually exceeded annual E. prolifera production. Therefore, novel extraction methods should be applied to increase the bioactive compounds of E. prolifera at the level of large-scale industrialization to support the efficient utilization of E. prolifera in aquafeeds. According to biological activities of E. prolifera, the growth, feed intake, lipid metabolism, antioxidant, immune parameters and stress resistance were investigated in Pacific white shrimp. It confirmed that bio-products of E. prolifera based on different processing procedures could be used to reduce the amount of E. prolifera in feed without compromising its biological function. It will helpful to improve the application of E. prolifera in aquaculture. A feeding trial was conducted to evaluate the biological activities of Enteromorpha prolifera bio-products in the diets of Pacific white shrimp (Litopenaeus vannamei). Bio-products of E. prolifera included E. prolifera meal, E. prolifera hydrolysate and E. prolifera polysaccharide, which was supplied using different processing procedures. The control diet was supplemented without any E. prolifera bio-products or dietary attractants. Experimental diets were formulated to contain 0.2% and 0.4% of E. prolifera hydrolysate (EPH0.2% and EPH0.4%), 0.03% of E. prolifera polysaccharide (EPP0.03%), 3% of E. prolifera meal (EPM3%), 0.1% of dimethyl-β-propiothetn (DMPT0.1%) and 1% of squid paste (SP1%). Shrimp (~8 g) were randomly distributed in 21 tanks and fed for 44 days. Feed intake showed that 3% of E. prolifera meal and 0.4% of E. prolifera hydrolysate in diets exhibited similar attractant effects as 0.1% of DMPT. Gross qualitative observation showed that the abundance of lipid droplets decreased in hepatopancreas of the EPH0.4% and EPM3% groups, which were supported by hepatopancreas triacylglycerol (TG), where significantly lower concentrations were observed in the EPH0.4% and EPM3% groups compared with the control group. Similarly, TG and low-density lipoprotein cholesterol in serum significantly decreased in the EPH0.4%, EPP0.03% and EPM3% groups compared to the control group. Supplemental E. prolifera bio-products resulted in significantly higher serum glutathione level of EPP0.03% or superoxide dismutase activities of EPH0.4% and EPM3%, but significantly lower serum malondialdehyde level of EPM3%. In addition, tnf-α expression in hepatopancreas was significantly down-regulated in shrimp fed the EPH0.2%, EPH0.4% and EPM3% diets. Based on survival analysis, E. prolifera bio-products improved the resistance of shrimp to hypoxic stresses. Thus, this study confirmed that bio-products of E. prolifera supplied using different processing procedures could be used to reduce the amount of E. prolifera in feed of shrimp without compromising their biological functions. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of imbalanced dietary BCAAs, especially Leu, on the growth and BCAA metabolism in turbot. A control diet was formulated by keeping optimum levels of Leu, Ile and Val. Four experimental diets were prepared by removing supplemental crystalline Leu (deficiency) or supplementing double the amount of Leu, Ile or Val (excess) in the control diet. The growth was not significantly decreased by an excess of Leu, Ile or Val. Fish fed an excess of any particular BCAA significantly increased its postprandial (2 and 6 h) concentration in the plasma, muscles, and liver, but did not decrease the other two BCAA concentrations. The expression of intestinal b0at1 was down-regulated by excessive dietary Leu, Ile or Val. For BCAA catabolism, the mRNA levels of bcat2 in the muscles as well as bckdha and bckdhb in the livers of the Leu-deficient group were the lowest among all the groups, but were up-regulated by excess dietary Leu, Ile or Val. In conclusion, in terms of growth, turbot had high plasticity to an excess of any particular BCAA. Meanwhile, the antagonistic effect caused by an excess of one BCAA were reflected in intestinal amino acid absorption and BCAA catabolism. [ABSTRACT FROM AUTHOR]

Simple Summary: The supply of fish oil, which provides long-chain polyunsaturated fatty acids (LC-PUFA) but is mostly wild-caught, is stagnating, resulting in limited availability and high prices. Therefore, many alternative lipid sources, mainly vegetable oils and rendered animal fats, are increasingly being applied in fish feeds. However, these lipid sources do not contain LC-PUFA, and therefore lower the fillet quality in terms of the LC-PUFA content. Feeding strategies such as alternate feeding between fish oil- and alternative oils-based diets have been applied to maintain the n-3 LC-PUFA contents in farmed fish. The present study validated the relevant efficacy. A 12-week feeding experiment was conducted to evaluate the effects of replacing fish oil (FO) with beef tallow (BT) on the fatty acid composition of farmed tiger puffer (Takifugu rubripes). Two replacement strategies were used: a standard Graded Dietary Replacement of FO with BT (GDR strategy) and Alternate Feeding between FO- and BT-based Diets (AFD strategy). The positive and negative control diets were formulated with 6% FO (FO-C group) or BT (BT-C group) as the sole added lipid source. In the GDR strategy, three experimental diets were formulated, with 25, 50 and 75% of the added FO in the FO-C diet replaced with BT, named 25BT, 50BT and 75BT, respectively. In the AFD strategy, alternated feeding patterns between the FO-C and BT-C diet—namely, 1, 2 and 3 weeks with BT-C followed by 1 week feeding with FO-C (1BT-1FO, 2BT-1FO and 3BT-1FO, respectively)—were applied. Each diet or feeding strategy was assigned to triplicate tanks. The results showed that dietary BT inclusion reduced the contents of long-chain polyunsaturated fatty acids (LC-PUFA) in both the muscle and liver (edible tissues for this species) of the experimental fish, and the liver displayed a more drastic decrease than the muscle. The LC-PUFA content linearly decreased with the decreasing dietary FO levels in the GDR strategy. However, in the AFD strategy, a linear relationship was not observed between the LC-PUFA content and the FO feeding duration. The 3BT-1FO treatment resulted in higher LC-PUFA content than 2BT-1FO. When comparing the two strategies with the same final FO administration level—namely, 50BT vs. 1BT-1FO, and in particular, 75BT vs. 3BT-1FO—the AFD strategy resulted in higher LC-PUFA contents in both the muscle and liver than the GDR strategy. In conclusion, when FO was replaced with BT in the diets, alternate feeding between FO- and BT-based diets resulted in a higher LC-PUFA content than the standard direct replacement. Three weeks of feeding with BT-C followed by one week of feeding with FO-C appeared to be a good alternate feeding pattern. This study provided a promising strategy of FO-sparing in fish farming when the LC-PUFA contents were maintained as high as possible. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the potential effect of graded levels of tryptophan on the growth, cannibalism, and 5-hydroxytryptpamine (5-TH) metabolism in pufferfish (Takifugu obscurus ♀ × Takifugu rubripes ♂). A 63-day feeding trial was performed wherein pufferfish were fed four diets. Three experimental diets were formulated with various levels of tryptophan based on the control diet. Four diets were named as T1, T2, T3, and T4, corresponding to 4.30, 7.80, 14.90, and 23.70 g kg−1 tryptophan of dry diet. Final body weight, weight gain, and specific growth rate were similar between the T1 and T4 groups, but exhibited a significantly increased trend compared to the T2 group. Although survival rate was not affected by various levels of dietary tryptophan, intraspecific cannibalism was significantly reduced in the group fed with highest level of tryptophan (T4). For free amino acid in brain, the concentration of tryptophan was the highest in the T3 group and the lowest in the T2 group, while phenylalanine, tyrosine, and methionine showed an opposite trend between those two groups. The levels of dietary tryptophan not only affected the expression of aromatic amino acid transporter TAT1, but also affected the expression of B0AT1, B0AT2, and 4F2hc in intestine, as well as B0AT1, y+LAT1, and LAT2 in brain. The activity of tryptophan hydroxylase (TPH) in serum increased with the increase of dietary tryptophan, and the expression of TPH1 in brain upregulated in the excessive tryptophan groups (T2, T3, and T4). MAO activity in serum as well as its gene expression in brain and intestine showed a decreased trend in the T4 group. In conclusion, excessive tryptophan (23.70 g kg−1 of dry diet, corresponding to 50.3 g kg−1 of dietary protein) in feed could mitigate cannibalistic behavior of pufferfish and promote the growth, and the reason for this effect might affect the metabolism of 5-TH in vivo. [ABSTRACT FROM AUTHOR]

The fatty acid accumulation in fish tissues are closely related to fish sex, yet the sex differences in fish fatty acids have not been fully known. This experiment investigated the sex differences in fatty acid composition of Chinese tongue sole (Cynoglossus semilaevis), which have typical sexual dimorphism and reversal. Three sexes, namely females, males and pseudomales, were involved in this study. Five batches of fish samples were collected from different fish farms in Shandong Province, China. These fish were identified to be at the gonadal development stage of III–IV. The disparity of fatty acid between sexes (female vs male and pseudomale vs. male) was evaluated by the distance coefficient (Dsex) and multi-variate similarity of percentages analysis (SIMPER). The highest Dsex values between females and males were found in gonad, followed by liver and eye. The fatty acids that differed between females and males were mainly DHA, 16:1n-7, 18:1n-9, and 18:0. The sex difference in DHA and 18:1n-9 contributed significantly to the overall sex difference in fatty acid of brain, eye, and gonad, whereas that in 16:1n-7 contributed significantly in heart, gill, liver, and muscle. The sex difference in 18:0 contributed significantly to the overall sex difference in fatty acid of heart, gill, gut, and skin. The sex differences in fatty acid between pseudomales and males were generally insignificant. In conclusion, at this gonadal development stage (III–IV), the sex differences in fatty acid between female and male Chinese tongue sole have emerged, mainly in the gonad. The testes had higher DHA levels but lower levels of 16:1n-7 and 18:1n-9 than ovaries. In addition, for pseudomales, the phenotypic sex more substantially determined the fatty acid composition than the genetic sex. [ABSTRACT FROM AUTHOR]

Cholesterol has been used as additive in fish feeds due to the reduced use of fish meal and fish oil. In order to evaluate the effects of dietary cholesterol supplementation (D-CHO-S) on fish physiology, a liver transcriptome analysis was performed following a feeding experiment on turbot and tiger puffer with different levels of dietary cholesterol. The control diet contained 30% fish meal (0% fish oil) without cholesterol supplementation, while the treatment diet was supplemented with 1.0% cholesterol (CHO-1.0). A total of 722 and 581 differentially expressed genes (DEG) between the dietary groups were observed in turbot and tiger puffer, respectively. These DEG were primarily enriched in signaling pathways related to steroid synthesis and lipid metabolism. In general, D-CHO-S downregulated the steroid synthesis in both turbot and tiger puffer. Msmo1, lss, dhcr24, and nsdhl might play key roles in the steroid synthesis in these two fish species. Gene expressions related to cholesterol transport (npc1l1, abca1, abcg1, abcg2, abcg5, abcg8, abcb11a, and abcb11b) in the liver and intestine were also extensively investigated by qRT-PCR. However, the results suggest that D-CHO-S rarely affected the cholesterol transport in both species. The protein-protein interaction (PPI) network constructed on steroid biosynthesis-related DEG showed that in turbot, Msmo1, Lss, Nsdhl, Ebp, Hsd17b7, Fdft1, and Dhcr7 had high intermediary centrality in the dietary regulation of steroid synthesis. In conclusion, in both turbot and tiger puffer, the supplementation of dietary cholesterol inhibits the steroid metabolism but does not affect the cholesterol transport. [ABSTRACT FROM AUTHOR]

Simple Summary: In some fish farming practices, cholesterol has been used as a feed additive for growth stimulation. Therefore, it is important to increase our knowledge and experience about dietary cholesterol effects on fish. This study provides useful information for the application of cholesterol in fish diets by analyzing the muscle lipidomics changes in turbot and tiger puffer in response to dietary cholesterol supplementation. Exogenous cholesterol has been supplemented into aqua-feeds due to the reduced proportions of fishmeal and fish oil. This study aimed to investigate the effects of dietary cholesterol supplementation on the muscle lipidomics of two marine fish species, turbot and tiger puffer. A 70-day feeding trial was conducted, where two low-fishmeal diets supplemented with 0 or 1% cholesterol were used. The lipidomic analysis with targeted tandem mass spectrometry showed that, in turbot, a total of 49 individual lipids exhibited significant differences in their abundance in response to dietary cholesterol, whereas the number was 30 for tiger puffer. Dietary cholesterol up-regulated the abundance of cholesterol and cholesterol ester in both species. In turbot, the dietary cholesterol also increased the abundance of triacylglycerol and acylcarnitine, whereas in tiger puffer, it primarily regulated the abundance of phospholipids and BMP. This was the first time the responses of marine fish muscle lipidomics to dietary cholesterol supplementation have been investigated. [ABSTRACT FROM AUTHOR]

Simple Summary: There have been reports on alternate feeding between diets based on different lipid sources, but how the intestinal microbiota of fish responds to alternate feeding strategies and its relevant roles in the host's health have not been reported. The present study provides novel results in this area. The main objective of the study was to assess how the intestinal microbiota of fish responds to alternate feeding between diets based on different lipid sources. Foremost, the juvenile turbot accepted alternate feeding between diets based on different lipid sources with no negative effects on survival or growth performance. Additionally, novel results were observed regarding the different bacterial compositions, microbial co-occurrence networks, and functional predictions among the different alternating feeding strategies. The results of this study indicate that a more comprehensive evaluation should be conducted from the perspective of intestinal microorganisms when a new feeding strategy is applied in aquaculture practices. These results will contribute to the nutritional regulation of aquatic animals from the perspective of microorganisms. A nine-week feeding trial was conducted to investigate changes in the intestinal microbiota of turbot in response to alternate feeding between terrestrially sourced oil (TSO)- and fish oil (FO)-based diets. The following three feeding strategies were designed: (1) continuous feeding with the FO-based diet (FO group); (2) weekly alternate feeding between soybean oil (SO)- and FO-based diets (SO/FO group); and (3) weekly alternate feeding between beef tallow (BT)- and FO-based diets (BT/FO group). An intestinal bacterial community analysis showed that alternate feeding reshaped the intestinal microbial composition. Higher species richness and diversity of the intestinal microbiota were observed in the alternate-feeding groups. A PCoA analysis showed that the samples clustered separately according to the feeding strategy, and among the three groups, the SO/FO group clustered relatively closer to the BT/FO group. The alternate feeding significantly decreased the abundance of Mycoplasma and selectively enriched specific microorganisms, including short-chain fatty acid (SCFA)-producing bacteria, digestive bacteria (Corynebacterium and Sphingomonas), and several potential pathogens (Desulfovibrio and Mycobacterium). Alternate feeding may maintain the intestinal microbiota balance by improving the connectivity of the ecological network and increasing the competitive interactions within the ecological network. The alternate feeding significantly upregulated the KEGG pathways of fatty acid and lipid metabolism, glycan biosynthesis, and amino acid metabolism in the intestinal microbiota. Meanwhile, the upregulation of the KEGG pathway of lipopolysaccharide biosynthesis indicates a potential risk for intestinal health. In conclusion, short-term alternate feeding between dietary lipid sources reshapes the intestinal microecology of the juvenile turbot, possibly resulting in both positive and negative effects. [ABSTRACT FROM AUTHOR]

Booming fish farming results in a relative shortage of fish oil (FO) supply, meaning that alternative oils are increasingly used in fish feeds, which leads to reduction of long-chain polyunsaturated fatty acids (LC-PUFAs) and other relevant changes in fish products. This study investigated the efficacy of an FO-finishing strategy in recovering the muscle quality of farmed tiger puffer. An eight-week feeding trial (growing-out period) was conducted with five experimental diets, in which graded levels (0 (control), 25, 50, 75, and 100%) of added FO were replaced by poultry oil (PO). Following the growing-out period was a four-week FO-finishing period, during which fish in all groups were fed the control diet. Dietary PO significantly decreased the muscle LC-PUFA content, whereas in general, the FO-finishing strategy recovered it to a level comparable with that of the group fed FO continuously. The recovery efficiency of EPA was higher than that of DHA. Dietary PO also led to changes of volatile flavor compounds in the muscle, such as butanol, pentenal, and hexenal, whereas the FO-finishing strategy mitigated the changes. In conclusion, the FO-finishing strategy is promising in recovering the LC-PUFA and volatile-flavor-compound composition in farmed tiger puffer after the feeding of PO-based diets. [ABSTRACT FROM AUTHOR]

The fish oil finishing (FOF) strategy, that is, re-feeding fish with fish oil (FO)-based diet after a certain period of feeding with alternative lipid source-based diets. On tiger puffer, the present study investigated the response of intestinal microbiota to FOF. Fish were fed four diets based on FO, soybean oil, palm oil and beef tallow as lipid sources, respectively, firstly for 50 days (growing-out period), and then fed the FO-based diet for 30 more days (FOF period). The results showed that dietary terrestrially sourced oils impaired the intestinal function in the growing-out period. However, the activities of amylase, trypsin and anti-oxidative enzymes (SOD, CAT, T-AOC), as well as gene expression of inflammatory cytokines (IL-1β, TNF-α, TGF-β) and tight junction protein (Claudin4, Claudin7, Claudin18, JAM, ZO-1) in the intestine were significantly recovered by FOF. The 16S rDNA sequencing analysis showed that FOF improved the similarity of bacterial community among the groups. The MetaStat analysis confirmed that FOF regulated the abundance of butyric acid-producing bacteria (Lachnospiraceae, Eubacterium, Butyricicoccus, Clostridium and Roseburia) and bacteria related to digestion and absorption (Sphingomonas, Romboutsia and Brevibacillus). In conclusion, FOF can recover the intestine function. The intestinal microbiota probably participated in and played a key role in the recovery process. [ABSTRACT FROM AUTHOR]

This study investigated the sex difference in fatty acid (FA) composition of six wild marine fish species, namely, Cleisthenes herzensteini, Platichthys bicoloratus, Pseudosciaena polyactics, Platycephalus indicus, Alosa sapidissima and Scomberomorus niphonius. The coefficient of distance value between sexes (Dsex) and multi‐variate similarity of percentages analysis (SIMPER) revealed universal existence of sex difference in FA composition, particularly in gonad, intestine and liver. Nonetheless, this sex difference was highly dependent on fish species. In general, DHA, 18:1n‐9, 16:1n‐7, 16:0 and EPA appeared to be the TOP FAs differentially abundant between sexes. [ABSTRACT FROM AUTHOR]

Myostatin (MSTN), a member of the transforming growth factor‐β (TGF‐β) superfamily, has retained the function of inhibiting muscle growth in fish. Turbot Scophthalmus maximus possesses two MSTN paralogs (MSTNa and MSTNb), but their respective response to dietary nutrients is still unclear. To clarify the effects of fish protein hydrolysate (FPH) on MSTNa and MSTNb, their mRNA levels were quantified in the muscle using the fasting–refeeding experiment and long‐term feeding trial. Gene expressions in various tissues showed that MSTNb but not MSTNa was expressed mostly in the muscle. After 2 weeks of starvation, MSTNa expression was up‐regulated in the muscle by refeeding FPH diet at 3 h and 6 h, while MSTNb expression was affected from 3 h to 3 days. Moreover, changes in MSTNb expression over time showed that significant reduction was observed at 3 h in fish refed optimal levels of FPH, but at 6 h in fish refed excessive levels of FPH. The expressions of two MSTN paralogs and related gene expressions of TGF‐β/SMAD‐dependent signalling pathway were not significantly altered by graded levels of FPH after 7 days of refeeding, while the expressions of MSTNb as well as the upstream FMODa and downstream SMAD4 were significantly affected after the long‐term feeding trial (8 weeks). In conclusion, FPH mainly modulated the expression of MSTNb in the muscle, whereas MSTNa was only affected in the early stage of refeeding. Furthermore, during the refeeding period, optimal levels of FPH in diets may regulate MSTNb more rapidly to initiate compensatory growth. [ABSTRACT FROM AUTHOR]

Peroxisome plays an important role in the modulation of lipid catabolism in fish. The present study investigated the tissue distribution of five peroxisomal fatty acid β‐oxidation genes, including vlcs, acox1, acox3, ehhadh and acaa1, in turbot and tiger puffer. Their expressions in response to different nutritional status (starvation and change in dietary lipid levels) were also studied. Both fish were fed diets with graded lipid levels (8%, 12% and 16%) for 9 weeks, followed by 1‐month of starvation. The five genes were highly expressed in the liver, intestine and pyloric caecum of turbot, and tiger puffer intestine. High‐lipid diets up‐regulated the expression of vlcs and acox3 in tiger puffer liver, whereas did not affect the expression of all genes investigated in turbot. During the starvation, expression of acox1, acox3 and acaa1 in turbot liver was increased firstly from day 0 to day 10, and then was decreased, while expression of vlcs in muscle, and acaa1 in muscle and subcutaneous adipose tissue was linearly increased. In tiger puffer liver, the expression of peroxisomal β‐oxidation genes increased linearly with starvation time, but the expression of acox3, ehhadh and acaa1 in the muscle firstly increased from day 0 to day 4–9, and thereafter decreased. In conclusion, the peroxisomal fatty acid β‐oxidation genes were highly expressed in the liver and intestine of both turbot and tiger puffer, and starvation could stimulate and high dietary lipid levels have less impact on their expression. [ABSTRACT FROM AUTHOR]

An 8-week feeding trial was conducted, where turbot were fed four experimental diets, containing different LPC levels (0%, 0.1%, 0.25%, and 0.5%, named LPC0, LPC0.1, LPC0.25, and LPC0.5, respectively). The intestinal morphology results showed that there were no widened lamina propria and mixed inflammatory cells in the LPC-supplemented groups. Dietary LPC remarkably decreased the expression of TLRs (TLR3, TLR8, TLR9, and TLR22), MyD88, and signaling molecules (NF-κB, JNK, and AP-1). Similarly, diets with LPC supplementation markedly depressed the gene expression of NF-κB and JNK signaling pathway downstream genes (TNF-α, IL-1β, Bax, Caspase9, and Caspase-3). Furthermore, dietary LPC modified the intestinal microbial profiles, increasing the relative abundance of short-chain fatty acids-producers, lactic acid bacteria, and digestive enzyme-producing bacteria. Predictive functions of intestinal microbiota showed that turbot fed LPC diets had a relatively higher abundance of functions, such as lipid metabolism and immune system, but a lower abundance of functions, such as metabolic diseases and immune system diseases. The activities of intestinal acid phosphatase and alkaline phosphatase were also increased by dietary LPC. In conclusion, LPC supplementation could regulate the intestinal mucosal barrier via the TLR signaling pathway and alter the intestinal microbiota profile of turbot fed high-lipid diets. [ABSTRACT FROM AUTHOR]

Booming fish farming results in relative shortage of fish oil (FO), making it urgent to explore alternative lipid sources. This study comprehensively investigated the efficacy of FO replacement with poultry oil (PO) in diets of tiger puffer (average initial body weight, 12.28 g). An 8-week feeding trial was conducted with experimental diets, in which graded levels (0, 25, 50, 75, and 100%, named FO-C, 25PO, 50PO, 75PO, and 100PO, respectively) of FO were replaced with PO. The feeding trial was conducted in a flow-through seawater system. Each diet was fed to triplicate tanks. The results showed that FO replacement with PO did not significantly affect the growth performance of tiger puffer. FO replacement with PO at 50-100% even slightly increased the growth. PO feeding also had marginal effects on fish body composition, except that it increased the liver moisture content. Dietary PO tended to decrease the serum cholesterol and malondialdehyde content but increase the bile acid content. Increasing levels of dietary PO linearly upregulated the hepatic mRNA expression of the cholesterol biosynthesis enzyme, 3-hydroxy-3-methylglutaryl-CoA reductase, whereas high levels of dietary PO significantly upregulated the expression of the critical regulatory enzyme of bile acid biosynthesis, cholesterol 7-alpha-hydroxylase. In conclusion, poultry oil is a good substitution for fish oil in the diets of tiger puffer. Poultry oil could replace 100% added fish oil in the diet of tiger puffer, without adverse effects on growth and body composition. [ABSTRACT FROM AUTHOR]

Throughout the evolution of fish nutrition science, the importance of dietary arachidonic acid (ARA, 20:4n‐6) has been somewhat overlooked, likely due to the polarizing research focus on docosahexaenoic (DHA, 22:6n‐3) and eicosapentaenoic (EPA, 20:5n‐3) acids. Nevertheless, especially during the past two decades, increasing attention has been paid to the requirements and functions of ARA in fish nutrition, and a number of relevant studies have now been conducted on a series of fish species, including both marine and freshwater species. Those studies revealed important and diverse functions of ARA in various physiological processes, such as growth, reproduction, stress resistance, pigmentation, immunity, lipid deposition and bone development. The results of the present review, in addition to providing a state‐of‐the‐art summary of current knowledge, also revealed the complexity of ARA functions in terms of dosages, species, development stage, sex and feeding period, as well as in terms of interaction between ARA and other nutritional or non‐nutritional factors. [ABSTRACT FROM AUTHOR]

Two 70-day feeding trials were conducted to reevaluate the necessity of cholesterol supplementation in diets of two important marine fish species, turbot and tiger puffer. Graded levels (0, 0.5, 1.0, 2.0, and 4.0% of dry matter) of cholesterol were added into the basal diet (with 30% fishmeal but no fish oil) to make five experimental diets, which were designated as control, CHO-0.5, CHO-1.0, CHO-2.0, and CHO-4.0, respectively. The dietary cholesterol concentration was measured to be 0.11%, 0.65%, 1.10%, 2.32%, and 4.59%, respectively. Each group had triplicate tanks, and each tank was stocked with 30 fish. The results showed that compared to the control, dietary cholesterol supplementation had no significant effect on the growth of both turbot and tiger puffer. Nevertheless, excess dietary cholesterol (CHO-2.0 and CHO-4.0) led to significant lower weight gain of tiger puffer compared to CHO-1.0. The feed intake decreased with increasing levels of dietary cholesterol. In general, the cholesterol supplementation decreased the crude lipid content in the liver, whereas 0.5% dietary cholesterol supplementation tended to increase the crude lipid content in the muscle of both species. Dietary cholesterol supplementation significantly regulated the lipid-related biochemical parameters in serum, liver, and muscle, and compared to turbot, tiger puffer lipid compositions had a higher buffering capacity in response to changes in dietary cholesterol level. Dietary cholesterol supplementation increased the 20:4n-6 content in the liver. Dietary cholesterol supplementation significantly downregulated the gene expression of HMG-COAr, upregulated the CYP7A1 expression, and downregulated the expression of lipid absorption and biosynthesis genes, but had no significant effect on gene expression of CPT1 and lipid transport-related genes (ApoA1, ApoA4, ApoB100, and ApoEα) in the liver. In conclusion, under the present experimental condition, dietary cholesterol supplementation had no significant effect on the growth of turbot and tiger puffer. In both species, dietary cholesterol supplementation significantly regulated the lipid accumulation and metabolism. [ABSTRACT FROM AUTHOR]

The relative shortage of fishmeal is challenging the sustainable development of aquaculture industry. A 56‐day feeding trial was conducted to evaluate the efficacy of fishmeal replacement by poultry by‐product meal (PBM) in juvenile obscure pufferfish. The basal diet with 45% (dry matter) fishmeal was used as the control diet, and in the four experimental diets PBM was added into the control diet to replace fishmeal at the level of 15% (PBM15), 30% (PBM30), 45% (PBM45) and 60% (PBM60) respectively. Each diet was fed to triplicate tanks, each of which was stocked with 30 fish. PBM30 did not significantly affect the growth performance, nutrient apparent digestibility coefficient, proximate composition, amino acid profile, muscle texture and serum biochemical indices (p > 0.05). PBM45 did not significantly (p > 0.05) affects the weight gain, but reduced the feed efficiency ratio (p < 0.05). PBM60 resulted in significantly reduced fish growth (p < 0.05). PBM‐supplemented diets resulted in lower contents of long chain‐polyunsaturated fatty acid in the liver, but higher deposition of C18:1n‐9 and C18:2n‐6 in experimental fish (p < 0.05). In conclusion, replacing 30% fishmeal with PBM in the diet did not compromise the growth performance, feed utilisation, health status and muscle texture of farmed obscure pufferfish. [ABSTRACT FROM AUTHOR]

Fatty acid-binding proteins (fabps) play important roles in lipid homeostasis. In the present study, 7 fabp isoforms, namely, fabp1, fabp2, fabp3, fabp4, fabp6, fabp7, and fabp10, in two marine teleosts, were characterized. In general, turbot and tiger puffer fabp genes showed high identity to their orthologs in other fish species and mammals, but tiger puffer Fabp6 shared the lowest identity to its known orthologs in zebrafish and human. The tissue distribution patterns of fabps were generally in accordance with their function features. However, tiger puffer fabps, in particular Fabp1, Fabp2, Fabp6 and Fabp7, may have functions distinct from other teleosts, as indicated by the phylogenetic tree and tissue distribution patterns. In both species, high dietary lipid levels downregulated the expression of fabp2, fabp3, fabp6, and fabp7a but tended to upregulate the fabp1 expression. Starvation downregulated the expression of most fabps in both fish species, but the downregulation of fabp expression in turbot was much more drastic and earlier compared to tiger puffer. Long-term (30-day) starvation increased the fabp7 expression in tiger puffer and tended to increase the fabp6 expression in turbot. Results of this study contribute to fish fabp physiology and its nutritional regulation. [ABSTRACT FROM AUTHOR]

A 56-day feeding trial was conducted in a flow-through seawater system to investigate the effects of lysophosphatidylcholine (LPC) on growth performance and lipid metabolism of turbot. Four experimental diets were prepared, differing only in the LPC supplementation, namely, 0 (LPC-0, control), 0.1 (LPC-0.1), 0.25 (LPC-0.25), and 0.5% (LPC-0.5) of dry matter. Each diet was randomly fed to triplicate tanks. LPC-0.1 and LPC-0.25 led to significantly higher weight gain than the control diet, and the highest weight gain was observed in LPC-0.1. Compared to the control group, the LPC-supplemented groups had higher survival and lower hepatosomatic index and viscerosomatic index. LPC-0.25 led to significantly lower contents of crude lipid and ash in whole fish. Dietary LPC supplementation led to a basic decrease in the lipid metabolism-related biochemical parameters in serum but had only very minor influence on the fatty acid composition in the liver and subcutaneous tissue around the fin (STF). High LPC levels upregulated the mRNA expression of BSAL and ApoEα in both the liver and STF. In conclusion, dietary LPC supplementation (0.1-0.25%) enhanced the growth, lowered the lipid accumulation in juvenile turbot, and significantly regulated the lipid metabolism. However, it seldom influenced the fatty acid composition. [ABSTRACT FROM AUTHOR]

The present study was aimed at screening suitable reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) in tiger puffer (Takifugu rubripes), an important aquaculture species in Asia and also a good model species for lipid research. Specifically, this reference gene screening was targeted at standardization of gene expression in different tissues (liver, muscle, brain, intestine, heart, eye, skin, and spleen) or under different nutritional conditions (starvation and different dietary lipid levels). Eight candidate reference genes (ribosomal protein L19 and L13 (RPL19 and RPL13), elongation factor-1 alpha (EF1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine guanine phosphoribosyl transferase1 (HPRT1), beta-2-Microglobulin (B2M), 18S ribosomal RNA (18SrRNA), and beta actin (ACTB)) were evaluated with four algorithms (geNorm, NormFinder, BestKeeper, and comparative ΔCt method). The results showed that different algorithms generated inconsistent results. Based on these findings, RPL19, EF1α, 18SrRNA, and RPL13 were relatively stable in different tissues of tiger puffer. During starvation conditions, ACTB/RPL19 was the best reference gene combination. Under different dietary lipid levels, ACTB/RPL13 was the most suitable reference gene combination. The present results will help researchers to obtain more accurate results in future qRT-PCR analysis in tiger puffer. [ABSTRACT FROM AUTHOR]

A 65‐day feeding trial was conducted to investigate the effects of different levels of dietary lipid on intestinal health of tiger puffer. Three experimental diets were formulated, differing in lipid level, that is, 90.3 (control‐suitable lipid, C‐SL), 130.3 (moderately high lipid, MHL) and 170.3 (extremely high lipid, EHL) g/kg dry matter. The results showed that with increasing dietary lipid levels, the activities of total antioxidant capacity, Na+ K+‐ATP, amylase and trypsin were significantly decreased by diets MHL and EHL. Compared with C‐SL, the mRNA expression of the intestinal tight junction proteins (Claudin14, Claudin18, junctional adhesion molecule‐A) were significantly down‐regulated and the serum diamine oxidase enzyme activity was significantly increased in the EHL group. Diet EHL up‐regulated the gene expression of the pro‐inflammatory cytokines, tumour necrosis factor‐α, interferon‐2, interleukin (IL)‐1β, IL‐8 and IL‐15, and down‐regulated that of anti‐inflammatory cytokine transforming growth factor ‐β. Sequencing of intestinal bacterial 16s rRNA V4 region showed that diet EHL significantly increased the Firmicutes/Bacteroidetes ratio, decreased the alpha diversity of intestinal microbiota and the relative abundance of some potential beneficial bacteria such as Lactobacillus, Sphingomonas and Thermus. These results showed that high level of dietary lipid (170.3 g/kg) had negative effects on function status, mucosal barrier and microbial communities of tiger puffer intestine, indicating that tiger puffer was sensitive to the dietary lipid level. [ABSTRACT FROM AUTHOR]

Use of terrestrially sourced oils (TSO) in the diet easily causes reduction in long‐chain polyunsaturated fatty acids (LC‐PUFA) in farmed fish. Alternate feeding between fish oil (FO)‐ and TSO‐based diets is a potential strategy for mitigating this LC‐PUFA reduction. The present study, with turbot, was aimed at comparing the fatty acid‐regulating effects of the alternate‐feeding strategy in different tissue types, namely, muscle, liver and subcutaneous tissue around the fin (STF). The control group were fed the FO‐based diet continuously for 13 weeks. Meanwhile, other groups were subjected to weekly alternation between the FO‐based diet and diets based on TSO, namely, linseed oil, soybean oil, rapeseed oil, palm oil and beef tallow, respectively. Each dietary group had triplicate tanks, and each tank was stocked with 35 fish. Tissue samples were collected at the end of Week 9 and 13. At Week 9, significant differences among dietary groups were observed in LC‐PUFA contents of all tissue types; at Week 13, the significant difference in muscle disappeared, but those in liver and STF remained. STF seemed more like a lipid storage site, with high contents of 18:1n‐9 and 18:3n‐3, but low LC‐PUFA contents. Among all the treatments, alternate feeding between FO‐ and beef tallow‐based diets resulted in the most similar fatty acid profile to the treatment with continuous FO feeding. This study added knowledge to the fatty acid composition of fish subcutaneous tissue, as well as to promising application of beef tallow in fish feed. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the effects of different dipeptides (lysine-leucine, lysine-glycine, and leucine-glycine) and free amino acids (lysine and leucine) on the growth, gene expression of intestinal peptide and amino acid transporters, and serum free amino acid concentrations in turbot. Fish (11.98 ± 0.03 g) were fed four experimental diets supplementing with crystalline amino acids (CAA), lysine-leucine (Lys-Leu), lysine-glycine (Lys-Gly), and leucine-glycine (Gly-Leu). Fish protein hydrolysate (FPH) containing a mixture of free amino acids and small peptides was designed as a positive control diet. There was no significant difference in the growth and feed utilization among three dipeptide diets (Lys-Leu, Lys-Gly, and Gly-Leu). Compared with the CAA group, feed efficiency ratio was significantly higher in the Lys-Leu and Lys-Gly groups, and protein efficiency ratio was significantly higher in the Lys-Leu group. For peptide transporter, oligopeptide transporter 1 (PepT1) mRNA level was not affected by dietary treatments. For amino acid transporters, lower expression of B0 neutral amino acid transporter 1 (B0AT1) and proton-coupled amino acid transporter 1 (PAT1) were observed in fish fed the dipeptide and FPH diets compared with the CAA diet. In conclusion, juvenile turbot fed Lys-Leu, Gly-Leu, and Lys-Gly had a similar growth performance, whereas lysine and leucine in the Lys-Leu form can be utilized more efficiently for feed utilization than those in free amino acid from. In addition, compared to free amino acids, dipeptides and fish protein hydrolysate in diets may down-regulate the expression of amino acid transporters but did not affect the expression of PepT1. [ABSTRACT FROM AUTHOR]

Keywords: arachidonic acid; characterization; cloning; Cynoglossus semilaevis; pmch2; sex dimorphism EN arachidonic acid characterization cloning Cynoglossus semilaevis pmch2 sex dimorphism 3472 3477 6 07/14/20 20200801 NES 200801 Melanin-concentrating hormone (MCH), originally discovered in the teleost pituitary, is a hypothalamic neuropeptide involved in the regulation of body colour in fish (Cerdá-Reverter & Canosa, 2009; Kawauchi, Kawazoe, Tsubokawa, Kishida, & Baker, 1983; Mizusawa, Kasagi, & Takahashi, 2018; Mizusawa, Kobayashi, Yamanome, Saito, & Takahashi, 2013; Mizusawa, Yamamura, Kasagi, Cerdá-Reverter, & Takahashi, 2018). The deduced Chinese tongue sole pMCH2 contains a 24-amino acid signal peptide (MISVSSVLITLVLSSVLSSHLVVVA) and a dibasic (RR) cleavage site near the C terminus that likely is used to generate the MCH peptides, a 23-amino acid peptide (NTELDMLRCMIGRVYRCWGTSN). The functions of MCH in fish reproduction and a potential gender difference in MCH action would merit investigation. Arachidonic acid, characterization, cloning, Cynoglossus semilaevis, pmch2, sex dimorphism. [Extracted from the article]

The tissue distribution pattern of lipid is highly diverse among different fish species. Tiger puffer has a special lipid storage pattern, storing lipid predominantly in liver. In order to better understand the lipid physiology in fish storing lipid in liver, the present study preliminarily investigated the tissue distribution of transcription for 29 lipid metabolism-related genes in tiger puffer, which are involved in lipogenesis, fatty acid oxidation, biosynthesis and hydrolysis of glycerides, lipid transport, and relevant transcription regulation. Samples of eight tissues, brain, eye, heart, spleen, liver, intestine, skin, and muscle, from fifteen juvenile tiger puffer were used in the qRT-PCR analysis. The intestine and brain had high transcription of lipogenic genes, whereas the liver and muscle had low expression levels. The intestine also had the highest transcription level of most apolipoproteins and lipid metabolism-related transcription factors. The transcription of fatty acid β-oxidation-related genes was low in the muscle. The peroxisomal fatty acid oxidation may dominate over mitochondrial β-oxidation in the liver and intestine of tiger puffer, and the MAG pathway probably predominates over the G3P pathway in re-acylation of absorbed lipids in the intestine. The intracellular glyceridases were highly transcribed in the brain, eye, and heart. In conclusion, in tiger puffer, the intestine could be a center of lipid metabolism whereas the liver is more likely a pure storage organ for lipid. The lipid metabolism in the muscle could also be inactive, possibly due to the very low level of intramuscular lipid. [ABSTRACT FROM AUTHOR]

Taurine (TAU) plays important roles in the metabolism of bile acids, cholesterol and lipids. However, little relevant information has been available in fish where TAU has been identified as a conditionally essential nutrient. The present study aimed to investigate the effects of dietary TAU on the metabolism of bile acids, cholesterol and lipids in tiger puffer, which is both an important aquaculture species and a good research model, having a unique lipid storage pattern. An 8-week feeding trial was conducted in a flow-through seawater system. Three experimental diets differed only in TAU level, that is, 1·7, 8·2 and 14·0 mg/kg. TAU supplementation increased the total bile acid content in liver but decreased the content in serum. TAU supplementation also increased the contents of total cholesterol and HDL-cholesterol in both liver and serum. The hepatic bile acid profile mainly includes taurocholic acid (94·48 %), taurochenodeoxycholic acid (4·17 %) and taurodeoxycholic acid (1·35 %), and the contents of all these conjugated bile acids were increased by dietary TAU. The hepatic lipidomics analysis showed that TAU tended to decrease the abundance of individual phospholipids and increase those of some individual TAG and ceramides. The hepatic mRNA expression study showed that TAU stimulated the biosynthesis of both bile acids and cholesterol, possibly via regulation of farnesoid X receptor and HDL metabolism. TAU also stimulated the hepatic expression of lipogenic genes. In conclusion, dietary TAU stimulated the hepatic biosynthesis of both bile acids and cholesterol and tended to regulate lipid metabolism in multiple ways. [ABSTRACT FROM AUTHOR]

The study aimed to investigate the effects of crystalline Lys and Leu, Lys‐Leu dipeptide and Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide, on growth, postprandial Lys and Leu concentrations, gene expression of peptide and amino acid (AA) transporters and related gene expression of protein synthesis pathway in turbot. Three diets (CAA, Di and Tri) contained Lys and Leu as free (Lys and Leu), dipeptide (Lys‐Leu) and tripeptide (Lys‐Leu‐Lys/Leu‐Lys‐Leu) forms, respectively. Improved growth was observed in the Di group compared with the CAA group. For peptide and AAs transporters, PepT1, B0AT1 and y+LAT2 mRNA levels were affected in proximal intestine by dietary treatments. Free Lys and Leu concentrations in the CAA group were significantly higher than that of the Di and Tri groups at 6 hr postfeeding in serum and at 2, 6 and 24 hr postfeeding in muscle. For protein synthesis pathway in muscle, Akt2, TOR and S6k1 gene expression were the highest in the Di group and the lowest in the CAA group, whereas MuRF1 relative expression was the highest in the Tri group. In conclusion, dietary Lys‐Leu dipeptide was utilized more efficiently than free Lys and Leu or Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide for turbot by regulating AAs transport, postprandial AAs concentration and the synthesis of muscle proteins. [ABSTRACT FROM AUTHOR]

The effects of fish protein hydrolysate (FPH) on growth, peptide and amino acid (AA) transporters, postprandial free AA and related gene expression of IGF‐1/AKT pathway were evaluated in turbot (Scophthalmus maximus). Three diets were formulated to contain the same low level of fishmeal; meanwhile 0, 45 and 180 g/kg FPH were, respectively, supplemented to the FF (FPH‐free), FL (FPH‐Low) and FH (FPH‐High) diets. Fish fed the FH diet improved the growth compared with the other groups. For peptide and AA transporters, PepT1, B0AT1, CAT1 and PAT1 mRNA levels in proximal or distal intestine decreased in fish fed the FH diet. The concentration of free total essential AAs in serum was higher in the FH treatment than that in the FF treatment at 2 and 6 hr after feeding. For IGF‐1/AKT pathway in muscle, IGF‐1, 4E‐BP1 and FoxO1 mRNA levels were the highest in the FH group, whereas IGF‐1R mRNA levels were the highest expression level in the FF group. In conclusion, down‐regulated AAs transport, alleviated the delayed postprandial peak of serum‐free AAs and increased muscle protein synthesis were observed to improve the growth when turbot was fed high FPH level diets containing a high plant protein. [ABSTRACT FROM AUTHOR]

Our previous studies have indicated that dietary arachidonic acid (ARA) significantly affects the gonadal steroidogenesis in the marine teleost tongue sole Cynoglossus semilaevis, and this effect was more positive in male fish than in female fish (Aquaculture, 468, 378–385). As a following up study, the present study was further aimed at investigating the possible mechanisms in the brain mediating the effects of dietary ARA on gonadal steroidogenesis. A 70‐day feed trial was repeated with two‐year‐old tongue sole, using three experimental diets with graded levels of ARA, 0.34%, 2.53%, and 9.63% of total fatty acids. Each diet was fed to triplicate groups of 23 fish (15 males and 8 females). The results confirmed the positive effect of dietary ARA on testosterone production in male fish. The concentration of gonadotrophin‐releasing hormone (GnRH) in serum responded to dietary ARA in a similar pattern with the testosterone concentration, but the concentration of gonadotrophin in serum was not affected by dietary ARA. The response of gene expression of PKCβ, ARRB1, ARRB2, ERK2 and ATF3 in the brain to dietary ARA was in good agreement with those of GnRH and testosterone, indicating the possible involvement of PKC‐ARRB‐ERK‐ATF3 pathway in signalling transduction of GnRH. However, the PI3K/Akt and TLR/NF‐κB pathways may not be directly involved in the regulation of GnRH metabolism by ARA. This is the first study reporting the possible involvement of PKC/MAPK pathways in regulation of reproductive endocrine processes by long‐chain polyunsaturated fatty acid in brain of marine fish. [ABSTRACT FROM AUTHOR]

A 74‐day feeding trial was conducted to investigate the efficacy of n‐6 polyunsaturated fatty acid (n‐6 PUFA) in diets for tiger puffer. Six experimental diets with graded levels of n‐6 PUFA (from 6.01 to 37.98 g/kg) replacing saturated fatty acids (SFAs) were used in this study. Increasing levels of n‐6 PUFA resulted in decreased weight gain (from 472.6% to 349.4%), feed efficiency and crude protein content in fish, but increased lipid accumulation in whole fish and liver. Liver samples from fish fed the lowest and highest level of n‐6 PUFA were subjected to transcriptomic assay. Analysis of the differentially expressed genes showed that high dietary levels of n‐6 PUFA tended to induce glycolysis and triacylglycerol biosynthesis, but reduce synthesis of amino acid and protein, and cause impairment to liver health. The mitochondrial DNA copy number, activity of mitochondrial respiratory chain complex I and glycogen content in the liver, which indicate energy supply status, significantly decreased with increasing levels of dietary n‐6 PUFA. In conclusion, high n‐6 PUFA/SFA ratios in diet led to reduced energy supply (by approximately 50%) and increased hepatic lipid accumulation (by 55%), which probably consequently negatively affect protein synthesis, liver health and growth of tiger puffer. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNA) have emerged as important regulators of lipid metabolism and have been shown to play multifaceted roles in controlling transcriptional gene regulation, but very little relevant information has been available in fish, especially in non-model fish species. With a feeding trial on a typical marine teleost tongue sole C. semilaevis followed by transcriptomic analysis, the present study investigated the possible involvement of lncRNA in hepatic mRNA expression in response to different levels of dietary DHA and EPA, which are two most important fatty acids for marine fish. An 80-day feeding trial was conducted in a flow-through seawater system, and in this trial three experimental diets differing basically in DHA/EPA ratio, i.e., 0.61 (D/E-0.61), 1.46 (D/E-1.46), and 2.75 (D/E-2.75), were randomly assigned to 9 tanks of experimental fish. A total of 124.04 G high quality genome-wide clean data about coding and non-coding transcripts was obtained in the analysis of hepatic transcriptome. Compared to diet D/E-0.61, D/E-1.46 up-regulated expression of 178 lncRNAs and 2629 mRNAs, and down-regulated that of 47 lncRNAs and 3059 mRNAs, while D/E-2.75 resulted in much less change in gene expression. The co-expression and co-localization analysis of differentially expressed (DE) lncRNA and mRNA among dietary groups were then conducted. The co-expressed DE lncRNA and mRNA were primarily enriched in GO terms such as Metabolic process, Intracellular organelle, Catalytic activity, and Oxidoreductase activity, as well as in KEGG pathways such as Ribosome and Oxidative phosphorylation. Overlap of co-expression and co-localization analysis, i.e., lncRNA–mRNA matches "XR_523541.1–solute carrier family 16, member 5 (slc16a5)" and "LNC_000285–bromodomain adjacent to zinc finger domain 2A (baz2a)," were observed in all inter-group comparisons, indicating that they might crucially mediate the effects of dietary DHA and EPA on hepatic gene expression in tongue sole. In conclusion, this was the first time in marine teleost to investigate the possible lncRNA–mRNA interactions in response to dietary fatty acids. The results provided novel knowledge of lncRNAs in non-model marine teleost, and will serve as important resources for future studies that further investigate the roles of lncRNAs in lipid metabolism of marine teleost. [ABSTRACT FROM AUTHOR]

A 74‐day feeding trial was conducted to evaluate the effects of dietary astaxanthin (ASTX) on antioxidative status, inflammatory response and microbiota in the intestine of tiger puffer. Four diets differed only in ASTX content, that is, 0 (Control), 50 (ASTX50), 100 (ASTX100) and 500 (ASTX500) mg/kg. Compared with Control, ASTX50 and ASTX100 significantly reduced the malondialdehyde content and spared the activities of antioxidative enzymes in hindgut; however, ASTX500 had higher activity and gene expression of antioxidative enzymes. Nuclear factor‐erythroid 2 p45‐related factor 2 (Nrf2) and Kelch‐like ECH‐associated protein 1 (Keap1) signalling were probably involved in the transcriptional regulation of antioxidative enzymes by dietary ASTX. Compared with Control and ASTX500, ASTX50 and ASTX100 decreased the leucocyte infiltration in the lamina propria of hindgut and down‐regulated the gene expression of tumour necrosis factor‐α (TNF‐α) and interleukin‐8 (IL‐8). ASTX markedly affected the overall structure of intestinal microbiota. The relative abundance of Arcobacter, which has harmful properties, was the lowest in ASTX100 but the highest in ASTX500. In conclusion, dietary ASTX at 50 or 100 mg/kg improved the antioxidative status, alleviated the inflammation and favourably modulated the microbiota in tiger puffer intestine, but excess ASTX exerted contrary effects. [ABSTRACT FROM AUTHOR]

The study was conducted to investigate the effects of taurine (Tau) alone or in combination with fish protein hydrolysate (FPH) on growth performance, the expression of Tau transporter (TauT) and metabolic profile in juvenile turbot. FM, FPH0, FPH0+T, FPH10 and FPH10+T diets, respectively, contained 300, 150, 150, 80, and 80 g/kg fishmeal. FPH10 and FPH10+T diets contained 62 g/kg FPH. FPH0+T and FPH10+T diets were, respectively, prepared by supplementing the FPH0 and FPH10 diet formulations with 8 g/kg Tau. Specific growth rate was the highest in FM group and the lowest in FPH10 group. TauT mRNA levels in fish fed Tau supplemented diets were significantly lower than that in Tau unsupplemented diets. NMR‐based metabolomics analysis showed that Tau contents in liver of FPH0+T and FPH10+T were significantly higher than that of FM, FPH0 and FPH10. In muscle, Tau contents were significantly decreased in the FPH10+T versus FPH0 and the FPH10+T versus FPH10 comparisons. In conclusion, 62 g/kg FPH to replace fishmeal may not affect Tau synthesis, transport and metabolism. However, Tau supplemented alone or in combination with a certain level of FPH could reduce the requirement for Tau synthesis and transport and increased Tau levels in muscle and liver. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract: The lipid‐regulating effects of astaxanthin (ASTX) have been widely reported in terrestrial animals. However, little relevant information has been available in fish although ASTX has been used as exogenous pigment and antioxidant in fish feed. A 74‐day feeding study was conducted to investigate the effects of dietary ASTX on lipid accumulation in the marine teleost tiger puffer. Four experimental diets differing only in ASTX supplementation, that is, 0 (control), 50 (ASTX50), 100 (ASTX100) and 500 (ASTX500) mg kg‐1, were randomly assigned to 12 tanks of juvenile tiger puffer. Compared to control, the liver lipid content in group ASTX50 was significantly higher, while those in groups ASTX100 and ASTX500 were lower. The muscle lipid contents in group ASTX500 were significantly higher compared to control. Group ASTX50 had the best growth performances, while diet ASTX500 seemed to have adverse effects. In the liver, compared to control, groups ASTX50 and ASTX100 showed significantly lower mRNA expressions of genes related to triglycerol synthesis and fatty acid synthesis, transport and uptake, but higher expressions of genes related to β‐oxidation and monoglycerol hydrolysis. In the muscle, compared to control, ASTX100 showed higher expressions of genes related to β‐oxidation. ASTX50 resulted in higher contents of saturated and monounsaturated fatty acids but lower contents of n‐3 polyunsaturated fatty acids in fish. In conclusion, astaxanthin in diets for tiger puffer differentially regulated the lipid accumulation in the liver and muscle, both in dose‐dependent manners. Excess dietary astaxanthin (500 mg/kg) had adverse effects on tiger puffer. [ABSTRACT FROM AUTHOR]

In this study, putative cDNA of three fatty acid transport protein ( FATP) isoforms, that is FATP1, FATP4 and FATP6, was cloned and characterized from the liver of Japanese seabass ( Lateolabrax japonicus), and their expression in response to diets with different arachidonic acid ( ARA) levels (0.05%, 0.22%, 0.37%, 0.60%, 1.38% and 2.32% of dry matter) was investigated through a feeding trial. Two subtypes of FATP1, that is FATP1a and FATP1b, were cloned for the first time. The Japanese seabass FATPs showed high identity to their orthologs in other fish species and mammals, but Japanese seabass FATP6 showed low identity to Japanese seabass FATP1 and FATP4. FATP1a gene was highly expressed in brain, liver and eye, whereas FATP1b had the highest gene expression in gill, followed by kidney, skin, eye, muscle and heart. FATP4 gene was primarily expressed in intestine, brain and eye. However, FATP6 had very low gene expression levels in almost all tissues. High levels of dietary ARA (0.60%~2.32%) enhanced the gene expressions of FATP1a and FATP4 in the intestine and the gene expression of FATP1a in the muscle, whereas the dietary ARA supplementation reduced the FATP1b mRNA expression in the liver. The gene expression of FATP1a, FATP4 and FATP6 in the liver, as well as the FATP4 gene expression in the muscle, was not significantly affected by dietary ARA levels. To our knowledge, this is the first study investigating the regulation of FATP gene expressions by dietary ARA. [ABSTRACT FROM AUTHOR]

A 1H NMR-based metabolomics approach was to explore the effect of ultrafiltered fish protein hydrolysate ( UF) levels on the liver and muscle metabolic profile of juvenile turbot ( Scophthalmus maximus L.). Fish protein hydrolysate ( FPH) was produced from by-products by enzymatic treatment, and UF was obtained by diluting FPH followed filtration. Fish were fed diets containing fish meal protein, which was, respectively, replaced by UF protein 0, 50, 100, 150 and 200 g kg−1 of dietary total protein ( UF-0, UF-5, UF-10, UF-15 and UF-20) for 68 days. OPLS- DA of liver and muscle showed that high levels of UF in diets may lead to corresponding tissue metabolites changes. In liver tissue, changing metabolites included dimethylamine, N,N-dimethylglycine, valine, isoleucine, leucine, arginine and alanine and were mainly involved in choline metabolism and amino acid metabolism. For muscle tissue, changing metabolites included lactate, alanine, proline, fumarate, tyrosine, histidine, cystathionine and taurine. And the metabolites were mainly involved in glycolysis and gluconeogenesis, proline metabolism and taurine metabolism. 1H NMR-based metabolomics is a useful approach to investigate different levels of UF on metabolic profile in liver and muscle tissues combined with the growth. [ABSTRACT FROM AUTHOR]

A 10-week feeding experiment was conducted to investigate the effects of different types of dietary protein hydrolysate ( PH) on growth performance, body composition, trypsin activity and serum transaminase of juvenile turbot. Four high plant protein diets contained different types of PH, fish PH ( FPH), yeast PH ( YPH), pig blood PH ( PBPH) and soy PH ( SPH), replacing 10% fishmeal of the basal diet. The basal diet with 30% fishmeal and no PH was used as the control diet (C). Each diet was assigned to triplicate groups of 30 fish. The specific growth rate ( SGR) was not significantly different between groups C and FPH, but groups C and FPH showed significantly higher SGR than other groups. The feed efficiency ratio ( FER) and protein efficiency ratio ( PER) were not significantly different among groups C, FPH and YPH, but groups PBPH and SPH showed significantly lower FER and PER than groups C and FPH. Group PBPH showed significantly higher hepatosomatic index than other groups except YPH. Fish fed YPH showed significantly lower whole-body protein content, but significantly higher whole-body lipid content than fish fed diets C, FPH and PBPH. The activities of serum GOT and GPT in group PBPH were higher than those in groups C, FPH and YPH. These results suggested that when used at a low level in high plant protein diets for juvenile turbot, FPH is a good alternative protein source and YPH also has the application potential, but PBPH and SPH can cause negative impacts on fish growth and health. [ABSTRACT FROM AUTHOR]

A 3-month feeding experiment was conducted in an in-door seawater system to investigate the effect of dietary vitamin E (Ve) on the sperm quality of turbot ( Scophthalmus maximus). D-α-tocopherol acetate was supplemented to the basal (control) diet (65.14 mg kg Ve) to obtain low and high levels of dietary Ve (244.60 mg kg, LVe; 721.60 mg kg, HVe). Compared with the control, sperm concentration was significantly increased in Ve-supplemented groups (LVe and HVe); while relative sperm volume and testis-somatic index were significantly increased in group HVe only. Sperm motility duration was significantly longer in group HVe than in the control, but no significant difference was observed in percent motility among groups. Sperm size, the uniformity of mitochondrial size, and the integrity of mitochondria cristae and plasma membrane were improved by dietary Ve, especially in HVe. The content of Ve in testis and liver as well as polyunsaturated fatty acids in sperm increased with dietary Ve. These results suggested that dietary Ve, especially at the high level (721.60 mg kg), significantly improved sperm concentration and motility duration and maintained normal sperm morphology of turbot. [ABSTRACT FROM AUTHOR]

Four experimental diets were fed to turbot to examine the effect of fish hydrolysate and ultra-filtered fish hydrolysate on growth performance, feed utilization and non-specific immune response. Fish hydrolysate was produced by enzymatic treatment and size fractionated using ultra-filtration ( UF). The permeate (molecular weight <1000 Da) after UF and the non-ultra-filtered fish hydrolysate ( NUF) were tested as feed ingredients. Diets UF1, UF2 contained 3.7%, 1.2% ultra-filtered fish hydrolysate to replace fish meal protein respectively. The diets UF1, NUF were identical in composition except that the molecular weight of fish hydrolysate in the diet. Fish meal was used in the control diet. All diets were made equal in protein, lipid and energy. Each experimental diet was fed to juvenile turbot (27.87 ± 0.04 g) in triplicate for 8 weeks. Results of this study indicate that the best overall growth and feed utilization of turbot juveniles were obtained with a diet containing higher dose of the small molecular weight compounds in fish hydrolysate. Acid phosphatase, alkaline phosphatase, lysozyme and superoxide dismutase activity in serum were not affected by diet. Total antioxidant capacity was improved with increasing level of low molecule weight fish hydrolysate ( UF1). [ABSTRACT FROM AUTHOR]

To compare the flavor components of shrimp Litopenaeus vannamei cultured in sea water and low salinity water, the chemical composition of muscle was analyzed, muscle extracts prepared, and their sensory components and sensory properties were assessed. Shrimp cultured in sea water had a higher content of crude protein, lower moisture, and a higher flesh pH compared to the low salinity samples ( P < 0.05), whereas no significant differences were found in crude lipid and ash. The free amino acid composition of muscle extracts and the amino acid composition of peptides from shrimp cultured in sea water and low salinity water were similar, that is, the former had a very high content of glycine, arginine, proline and alanine, and the latter a very high content of glutamate, glycine, arginine, proline and alanine. The major components of nucleotides were adenosine monophosphate and inosine monophosphate, and the major components of organic acids were acetic acid and malic acid. Extracts from shrimp cultured in sea water had enhanced umami, sweetness and overall flavor, and less of an earthy-musty taste compared to samples cultured in low salinity water ( P < 0.05). Aftertaste did not differ between the two shrimp extracts ( P > 0.05). [ABSTRACT FROM AUTHOR]