Effects of essential amino acids supplementation in a low-protein diet on growth performance, intestinal health and microbiota of juvenile blotched snakehead (Channa maculata).

Developing a low-protein feed is important for the sustainable advancement of aquaculture. The aim of this study was to investigate the effects of essential amino acid (EAA) supplementation in a low-protein diet on the growth, intestinal health, and microbiota of the juvenile blotched snakehead, Channa maculata in an 8-week trial conducted in a recirculating aquaculture system. Three isoenergetic diets were formulated to include a control group (48.66 % crude protein (CP), HP), a low protein group (42.54 % CP, LP), and a low protein supplementation EAA group (44.44 % CP, LP-AA). The results showed that significantly lower weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER), and feed efficiency ratio (FER) were observed in fish that were fed LP than in the HP and LP-AA groups (P < 0.05). The HP and LP-AA groups exhibited a significant increase in intestinal villus length, villus width, and muscular thickness compared to the LP group (P < 0.05). Additionally, the HP and LP-AA groups demonstrated significantly higher levels of intestinal total antioxidant capacity (T-AOC), catalase (CAT), and superoxide dismutase (SOD) and lower levels of malondialdehyde (MDA) compared to the LP group (P < 0.05). The apoptosis rate of intestinal cells in the LP group was significantly higher than those in the LP and HP groups (P < 0.05). The mRNA expression levels of superoxide dismutase (sod), nuclear factor kappa B p65 subunit (nfκb-p65), heat shock protein 70 (hsp70), and inhibitor of NF-κBα (iκba) in the intestine were significantly higher in the LP group than those in the HP and LP-AA groups (P < 0.05). The 16s RNA analysis indicated that EAA supplementation significantly increased the growth of Desulfovibrio and altered the intestinal microflora. The relative abundances of Firmicutes and Cyanobacteria were positively correlated with antioxidant parameters (CAT and T-AOC), whereas Desulfobacterota was negatively correlated with sod and T-AOC. The genera Bacillus, Bacteroides, and Rothia were associated with the favorable maintenance of gut health. In conclusion, dietary supplementation with EAAs to achieve a balanced amino acid profile could potentially reduce the dietary protein levels from 48.66 % to 44.44 % without adversely affecting the growth and intestinal health of juvenile blotched snakeheads.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues / Respostas fisiológicas de tambaqui (Colossoma macropomum) alimentado com dietas suplementadas com silagem de resíduos de pescado e vegetais

The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

A procura por insumos que substituam produtos de alto custo, como óleo e farinha de peixe, na fabricação de rações para uso na aquicultura é crescente, sendo necessário garantir a qualidade nutricional das dietas. A silagem produzida a partir de resíduos de pescado e de vegetais apresenta-se como uma alternativa de baixo custo e eficiente fonte proteica. O objetivo do presente estudo foi avaliar as respostas fisiológicas e bioquímicas de alevinos de tambaqui alimentados com quatro níveis de inclusão de silagem em ração comercial com 28% de proteína bruta, em dois períodos: 45 e 90 dias. Cada tratamento foi realizado em três repetições, com 10 tambaquis em cada caixa experimental de 100 L. Ao término de cada período estabelecido, amostras sanguíneas foram coletadas de cinco animais de cada repetição para determinação das variáveis hematológicas e bioquímicas. Peso, comprimento total e índices hepatossomático e lipossomático de espécimes alimentados com silagem não mostraram alterações significativas em ambos os períodos de avaliação, bem como os valores de hematócrito. A concentração de hemoglobina de tambaqui após 45 dias de alimentação aumentou quando foi fornecida dieta com inclusão de 20% de silagem. A contagem de eritrócitos, volume corpuscular médio e hemoglobina corpuscular média não apresentaram alterações entre os tratamentos, em ambos os períodos. As proteínas totais aumentaram significativamente no plasma de tambaquis que receberam dietas com inclusão de 5 e 10% de silagem, avaliados após 45 dias de alimentação. Evidenciou-se que os grupos com inclusão de silagem na dieta não apresentaram alterações significativas nos parâmetros avaliados, assim não sendo prejudiciais à higidez do tambaqui. Portanto, o uso da silagem como suplemento alimentar durante o cultivo do tambaqui é uma alternativa sustentável para produtores, por promover a redução do descarte de resíduos de pescado e da agricultura.

Phytogenic Bioactive Compounds Shape Fish Mucosal Immunity.

Aquaculture growth will unavoidably involve the implementation of innovative and sustainable production strategies, being functional feeds among the most promising ones. A wide spectrum of phytogenics, particularly those containing terpenes and organosulfur compounds, are increasingly studied in aquafeeds, due to their growth promoting, antimicrobial, immunostimulant, antioxidant, anti-inflammatory and sedative properties. This trend relies on the importance of the mucosal barrier in the fish defense. Establishing the phytogenics' mode of action in mucosal tissues is of importance for further use and safe administration. Although the impact of phytogenics upon fish mucosal immunity has been extensively approached, most of the studies fail in addressing the mechanisms underlying their pharmacological effects. Unstandardized testing as an extended practice also questions the reproducibility and safety of such studies, limiting the use of phytogenics at commercial scale. The information presented herein provides insight on the fish mucosal immune responses to phytogenics, suggesting their mode of action, and ultimately encouraging the practice of reliable and reproducible research for novel feed additives for aquafeeds. For proper screening, characterization and optimization of their mode of action, we encourage the evaluation of purified compounds using in vitro systems before moving forward to in vivo trials. The formulation of additives with combinations of compounds previously characterized is recommended to avoid bacterial resistance. To improve the delivery of phytogenics and overcome limitations associated to compounds volatility and susceptibility to degradation, the use of encapsulation is advisable. Besides, newer approaches and dedicated methodologies are needed to elucidate the phytogenics pharmacokinetics and mode of action in depth.

Exploring the garlic (Allium sativum) properties for fish aquaculture.

The aquaculture industry's rapid growth to meet commercial demand can trigger an outbreak of infectious diseases due to high-density farming. Antibiotic overuse and misuse in fish farming and its global health consequences have led to searching for more natural alternatives such as medicinal plants. In this sense, garlic (Allium sativum) has different bioactive compounds with biological properties for animal health. Among them are the ajoene, alliin, and allicin, which confer biological properties such as growth promotion, antimicrobial, antiviral, antioxidant, and antiparasitic. Ways to use garlic in aquaculture include oil, fresh mash, aqueous extract, and garlic powder. The powder presentation is the most used in aquaculture; it is generally applied by oral administration, adding to the feed, and the dose used ranges from 0.05 to 40 g/kg of feed. Garlic has been used in the aquaculture of different species such as rainbow trout (Oncorhynchus mykiss), spotted grouper (Epinephelus coioides), catfish (Clarias gariepinus), tilapia (Oreochromis niloticus), guppy fish (Poecilia reticulata), goldfish (Carassius auratus), and barramundi (Lates calcarifer). In addition to its properties, garlic's usage became popular, thanks to its low cost, easy incorporation into food, and little environmental impact. Therefore, its application can be an effective solution to combat diseases, improve organisms' health using natural supplies, and as an alternative to antibiotics. This review reports and discusses plant-derived products' beneficial properties, emphasizing garlic and its usages in fish aquaculture.

Digested soybean protein and taurine influence bile acid level, lipase activity, lipid digestibility, and growth performance of pompano (Trachinotus blochii).

This study aimed to examine the effects of dietary digested soybean protein (DSP) and taurine on bile acid (BA) level, lipase activity, lipid apparent digestibility coefficient (ADC), and growth performance of pompano (Trachinotus blochii). Five diets were formulated with fish meal (FM), defatted soybean meal (SBM), and the DSP as main dietary protein sources. The diets were denoted as follows: FMD (FM-based diet), SBMD (SBM-based diet), SBM+TD (SBM-based diet plus taurine), DSPD (DSP-based diet), and DSP+TD (DSP-based diet plus taurine). Fingerling pompano with an initial body weight (BW) of 21.4 g were stocked in 500-L tanks, with triplicate tanks per dietary treatment. For 8 weeks, the fish were hand-fed the experimental diets to apparent satiation twice daily. The results showed that the DSPD and DSP+TD groups had significantly higher final BW, weight gain, and specific growth rate, but lower feed conversion ratio, than the SBMD and SBM+TD groups, respectively (P < 0.05). There were no significant differences in growth and feed performances between fish fed DSP+TD and FMD. The gallbladder and anterior intestinal BA levels, anterior intestinal lipase activity, and lipid and protein ADCs were markedly increased in fish fed DSPD and DSP+TD compared to those fed SBMD (P < 0.05), and no significant differences were detected between the DSP+TD and FMD groups. The findings of the present study suggested that dietary DSP inclusion with taurine supplementation might effectively improve lipid digestion and this contributed to growth enhancement in pompano fed a soybean protein-based diet.

Tributyrin Plays an Important Role in Regulating the Growth and Health Status of Juvenile Blunt Snout Bream (Megalobrama amblycephala), as Evidenced by Pathological Examination.

The present study aimed to assess the role of tributyrin (TB) in regulating the growth and health status of juvenile blunt snout bream (Megalobrama amblycephala) through an 8-week feeding experiment. Six groups were fed experimental diets with added TB percentages of 0% (control group), 0.03%, 0.06%, 0.09%, 0.12% and 0.15%. The present results showed that TB supplementation in feed had some positive impacts on FW, WG, FCR and SGR, and the best results were found in the 0.06% TB group (P<0.05). However, TB supplementation in feed had no significant effects on SR, CF, VSI or whole-body composition (P>0.05). TB supplementation in feed increased antioxidant capacity and immunological capacity and attenuated the inflammatory response by increasing the activity of T-SOD, GPx, CAT and the levels of anti-inflammatory cytokines (IL-10 and TGF-ß) and decreasing the levels of MDA and anti-inflammatory cytokines (TNF-α) (P<0.05). Furthermore, TB supplementation improved immunity by increasing the levels of immunoglobulins (IgM and IgG), C3 and IFN-γ (P<0.05). Surprisingly, 0.06%-0.12% TB supplementation significantly increased the content of IL-1ß (P<0.05). However, TB supplementation in feed had no significant effects on the plasma content of GSH, HSP70, IL-8 and the activity of T-AOC (P>0.05). The possible mechanism was that TB activated PI3K/Akt/Nrf2 and inhibits the NF-κB signaling pathway, further regulating the mRNA levels of key genes with antioxidant capacity and the inflammatory response; for example, it increased the mRNA levels of Nrf2, Cu/Zn-SOD, HO-1, CAT, Akt, PI3K, GPx, IL-10, and TGF-ß and decreased the mRNA levels of NF-κB and TNF-α (P<0.05). In addition, 0.06%-0.15% TB supplementation significantly increased the mRNA levels of IL-1ß (P<0.05). TB supplementation in feed had no significant effects on the mRNA levels of HSP70, Mn-SOD and IL-8 (P>0.05). Evidence was presented that TB supplementation decreased the mortality rate caused by Aeromonas hydrophila challenge. In pathological examination, TB supplementation prevented hepatic and intestinal damage. Generally, TB supplementation improved the growth performance of juvenile blunt snout bream. Furthermore, TB supplementation activated PI3K/Akt/Nrf2 and inhibited the NF-κB signaling pathway, regulating health status and preventing hepatic and intestinal damage.

Comparative genomic analysis of human GLI2 locus using slowly evolving fish revealed the ancestral gnathostome set of early developmental enhancers.

BACKGROUND: The zinc finger-containing transcription factor Gli2, is a key mediator of Hedgehog (Hh) signaling and participates in embryonic patterning of various organs including the central nervous system (CNS) and limbs. Abnormal expression of Gli2 can impede the transcription of Hh target genes through disruption of proper balance between Gli2 and Gli3 functions. Therefore, delineation of enhancers that are required for complementary roles of Glis would allow the interrogation of those pathogenic variants that cause gene dysregulation, and a corresponding abnormal phenotype. Previously, we reported tissue-specific enhancers for Gli family including Gli2 through direct tetrapod-teleost comparisons. RESULTS: Here, we employed the sequence alignments of slowly evolving spotted gar and elephant shark and have identified six novel conserved noncoding elements in human GLI2 containing locus. Zebrafish-based transgenic assays revealed that combined action of these autonomous CNEs reflects many aspects of Gli2 specific endogenous transcriptional activity, including CNS and pectoral fins. CONCLUSION: Taken together with our previous findings, this study suggests that Hh-signaling controlled deployment of Gli2 activity in embryonic patterning arose in the common ancestor of gnathostomes. These GLI2 specific cis-regulatory modules will help to identify DNA variants that probably reside outside of coding intervals and are associated with congenital anomalies.

Properties of Carotenoids in Fish Fitness: A Review.

Carotenoids, one of the most common types of natural pigments, can influence the colors of living organisms. More than 750 kinds of carotenoids have been identified. Generally, carotenoids occur in organisms at low levels. However, the total amount of carotenoids in nature has been estimated to be more than 100 million tons. There are two major types of carotenoids: carotene (solely hydrocarbons that contain no oxygen) and xanthophyll (contains oxygen). Carotenoids are lipid-soluble pigments with conjugated double bonds that exhibit robust antioxidant activity. Many carotenoids, particularly astaxanthin (ASX), are known to improve the antioxidative state and immune system, resulting in providing disease resistance, growth performance, survival, and improved egg quality in farmed fish without exhibiting any cytotoxicity or side effects. ASX cooperatively and synergistically interacts with other antioxidants such as α-tocopherol, ascorbic acid, and glutathione located in the lipophilic hydrophobic compartments of fish tissue. Moreover, ASX can modulate gene expression accompanying alterations in signal transduction by regulating reactive oxygen species (ROS) production. Hence, carotenoids could be used as chemotherapeutic supplements for farmed fish. Carotenoids are regarded as ecologically friendly functional feed additives in the aquaculture industry.

A fine-scale multi-step approach to understand fish recruitment variability.

Recruitment is one of the dominant processes regulating fish population productivity. It is, however, notoriously difficult to predict, as it is the result of a complex multi-step process. Various fine-scale drivers might act on the pathway from adult population characteristics to spawning behaviour and egg production, and then to recruitment. Here, we provide a holistic analysis of the Northwest Atlantic mackerel recruitment process from 1982 to 2017 and exemplify why broad-scale recruitment-environment relationships could become unstable over time. Various demographic and environmental drivers had a synergetic effect on recruitment, but larval survival through a spatio-temporal match with prey was shown to be the key process. Recruitment was also mediated by maternal effects and a parent-offspring fitness trade-off due to the different feeding regimes of adults and larvae. A mismatch curtails the effects of high larval prey densities, so that despite the abundance of food in recent years, recruitment was relatively low and the pre-existing relationship with overall prey abundance broke down. Our results reaffirm major recruitment hypotheses and demonstrate the importance of fine-scale processes along the recruitment pathway, helping to improve recruitment predictions and potentially fisheries management.

Protection of Siganus oramin, rabbitfish, from heavy metal toxicity by the selenium-enriched seaweed Gracilaria lemaneiformis.

Seaweed is an inherently important entity in marine ecosystems. It is not only consumed by aquatic animals but also improves environmental quality in the mariculture. Seaweed is also part of the diet of human beings. The purpose of the present study was to evaluate the antagonism of selenium (Se)-enriched Gracilaria lemaneiformis against heavy metals, specifically, the potential of dietary Se-enriched Gracilaria to protect against heavy metal toxicity in rabbitfish (Siganus oramin). Growth rate, heavy metal (Se, Cd, Pb, Cu, Zn and Cr) concentrations, malondialdehyde (MDA), metallothionein (MT), and the activity of the antioxidants, glutathione peroxidase (GPX), catalase (CAT) and superoxide dismutase (SOD) were all assessed. The results showed that the total organic and inorganic Se concentration for the 250 mg L-1 Se-enriched Gracilaria was significantly higher than those of the 50 and 10 mg L-1 treatments after 3 days of enrichment. The mean total Se concentrations in Gracilaria were 42.5 µg g-1 in the 250 mg L-1 treatment, 13.5 µg g-1 in the 50 mg L-1 treatment and 2.5 µg g-1 in the 10 mg L-1 treatment, respectively. Organic Se accounts for 80-82% of total Se in Se-enriched Gracilaria. The Se concentration of rabbitfish fed Se-enriched Gracilaria was significantly higher than control. Furthermore, Se increased Cu and Zn absorption, and enhanced MT generation, and improved GPX, CAT, and SOD antioxidant activity, and decreased MDA concentrations and lipid peroxidation levels, all antagonistic to Cd, Pb and Cr. The effects of Se-enriched Gracilaria on waterborne Cd, Pb and Cr-induced toxicity occurred via both enzymatic and non-enzymatic antioxidative mechanisms in rabbitfish. Selenium had synergistic effects on Zn and Cu in rabbitfish. For the 50 mg L-1 Se-enriched Gracilaria treatment, the Se, Cu, Zn, and antagonistic Cd, Pb, Cr, and the antioxidant enzymes CAT, SOD, GPX activities, and MT concentrations in rabbitfish were higher than that with the 250 mg L-1 and 10 mg L-1 Se-enriched Gracilaria treatments. The 50 mg L-1 Se treatment of Gracilaria was deemed to be the optimum concentration to promote growth of rabbitfish. Therefore, the obtained results suggest Se-enriched Gracilaria can antagonize heavy metal toxicity, and is an advisable Se supplement to improve the edible safety of cultured animals.

The effects of exposure to crude oil or PAHs on fish swim bladder development and function.

The failure of the swim bladder to inflate during fish development is a common and sensitive response to exposure to petrochemicals. Here, we review potential mechanisms by which petrochemicals or their toxic components (polycyclic aromatic hydrocarbons; PAHs) may affect swim bladder inflation, particularly during early life stages. Surface films formed by oil can cause a physical barrier to primary inflation by air gulping, and are likely important during oil spills. The act of swimming to the surface for primary inflation can be arduous for some species, and may prevent inflation if this behavior is limited by toxic effects on vision or musculature. Some studies have noted altered gene expression in the swim bladder in response to PAHs, and Cytochrome P450 1A (CYP1A) can be induced in swim bladder or rete mirabile tissue, suggesting that PAHs can have direct effects on swim bladder development. Swim bladder inflation failure can also occur secondarily to the failure of other systems; cardiovascular impairment is the best elucidated of these mechanisms, but other mechanisms might include non-inflation as a sequela of disruption to thyroid signaling or cholesterol metabolism. Failed swim bladder inflation has the potential to lead to chronic sublethal effects that are as yet unstudied.

Effects of dietary asafoetida (Ferula sinkiangensis K. M. Shen) levels on feeding attraction activity, growth performance, healthiness, and digestive enzyme activity in juvenile Lateolabrax japonicus.

The study was to investigate effects of asafoetida (Ferula sinkiangensis K. M. Shen) powder on feeding attraction activity (FAA), growth performance, healthiness, and digestive enzyme activity of juvenile Lateolabrax japonicus. Six concentration levels (0, 5, 10, 15, 20, and 25 g/kg diets) were formulated for luring and feeding experiment. Results showed that asafoetida could stimulate the appetite of L. japonicus at the dietary levels from 10 to 25 g/kg; reduce the feed conversion ratio (FCR) and feed intake (FI) at 10-20 g/kg; increase the weight gain (WG) and specific growth rate (SGR) at 5-10 g/kg; increase the hepatosomatic index (HSI), body crude lipid content, serum total protein (TP) content, and lysozyme activity at 10-15 g/kg; decrease the moisture at 10-15 g/kg; and increase the serum total superoxide dismutase (T-SOD) activity at 5-15 g/kg, when compared with the control group (p < 0.05). Digestive enzyme activities including amylase (AMS) and trypsin in the intestine were significantly affected by the asafoetida powder (p < 0.05). Regression analyses between the FAA, FCR, WG, SGR, HSI, LZM, T-SOD, AMS, trypsin, and the dietary asafoetida powder levels showed that the optimal additional amount was 16.95, 13.65, 8.36, 8.15, 15.45, 9.94, 8.75, 11.77, and 7.07 g/kg, respectively, indicating that the optimal amount of asafoetida powder was located in 7.07-16.95 g/kg diet. However, combined with the significant difference analyses obtained from the current study, we would suggest the additive suitable dosage was 10 g/kg.

Effects of dietary n-3 highly unsaturated fatty acids levels on growth, lipid metabolism and innate immunity in juvenile golden pompano (Trachinotus ovatus).

To investigate the effects of dietary n-3 highly unsaturated fatty acids (HUFA) levels on growth, lipid metabolism and innate immunity in juvenile golden pompano Trachinotus ovatus, a marine carnivorous teleost, a total of 450 fish (average body weight: 14.84 g) were randomly distributed into 18 cages at sea, each dietary group with three cages and respectively fed six diets (D1-D6) with 2.30% (D1), 0.64% (D2), 1.00% (D3), 1.24% (D4), 1.73% (D5), or 2.10% (D6) n-3 HUFA. Here, D1 with fish oil as lipid source was set as control, while D2-D6 used a mixed vegetable oil as lipid source and supplemented with docosahexaenoic acid- (DHA) and eicosapentaenoic acid- (EPA) enriched oils to adjust the n-3 HUFA levels. After 8 weeks feeding, the daily growth coefficient (DGC), specific growth rate (SGR) and feed efficiency ratio (FER) showed no significant difference among the six dietary groups (P > 0.05). The levels of EPA and DHA in serum and liver increased with the dietary n-3 HUFA levels. The activity of total superoxide disumutase (T-SOD) in serum of fish fed D4 and D5 were significantly higher than that of the other groups, whereas the opposite was true for serum IL-1ß and IL-6 levels as well as liver malondialdehyde (MDA) content. The mRNA levels of genes related to hepatic lipid metabolism including sterol regulatory element-binding protein-1 (srebp-1), fatty acid binding protein 1 (fabp1), peroxisome proliferators-activated receptor alpha (pparα), elongase of very long-chain fatty acids 5 (elovl5) and fatty acyl desaturase 2 (fads2) were down-regulated in fish fed the diets with high n-3 HUFA levels, while those of apolipoprotein b 100 (aprob 100) and carnitine palmitoyl transferase 1 (cpt1) increased significantly as increasing n-3 HUFA levels up to 1.73% (D2-D5), but decreased in the 2.10% n-3 HUFA group (D6). In addition, the expression levels of genes related to innate immunity including interleukin-10 (il-10) and transforming growth factor ß1 (tgf-ß1) increased significantly when dietary n-3 HUFA increased from 0.64% to 1.73%, whereas the opposite was true for the expression levels of nuclear factor kappa-B (nf-κb), interleukin-1ß (il-1ß), interleukin-6 (il-6) and interleukin-8 (il-8). Overall, the results indicated that dietary n-3 HUFA at 1.24-1.73% (D4-D5) can effectively improve fatty acid profiles, lipid metabolism, antioxidant capacity and immune response of golden pompano.

Effects of lysine and leucine in free and different dipeptide forms on the growth, amino acid profile and transcription of intestinal peptide, and amino acid transporters in turbot (Scophthalmus maximus).

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.

Melanocortin-4 receptor regulation of reproductive function in black rockfish (Sebastes schlegelii).

Melanocortin-4 receptor (MC4R) is a G protein-coupled receptor with multiple functions in mammals. However, the functions of MC4R in fish have not been investigated extensively. The purpose of this study was to determine potential regulation of reproduction by the MC4R. We cloned the black rockfish MC4R and analyzed its tissue distribution and function. The results showed that black rockfish mc4r cDNA consisted of 981 nucleotides encoding a protein of 326 amino acids. The quantitative PCR data showed that mc4r mRNA was primarily expressed in the brain, gonad, stomach and intestine. In the brain, mc4r was found to be primarily located in the hypothalamus. Both α-MSH and ß-MSH increased gnih expression and decreased sgnrh and cgnrh expression (P < 0.05). α-MSH and ß-MSH had opposite effects on kisspeptin expression. In contrast, α-MSH and ß-MSH increased the expression of cyp11, cyp19, 3ß-hsd and star. In summary, our study shows that MC4R in black rockfish might regulate reproductive function and that the effects of α-MSH and ß-MSH might differ.

Evaluating the lucrative role of probiotics in the aquaculture using microscopic and biochemical techniques.

The present study focused to elucidate the promising role of probiotics in the fish culture, by employing microscopic and biochemical parameters to rationalize the possible outcomes. After preliminary feeding trial histopathological analysis revealed clear and visible changes in the intestinal and liver cells of CTRL (control group) compared to probiotics supplemented group, the bacterial inoculations resulted in shorten and swelled villi, while liver kupffer cells increased many folds in the CTRL. Moreover, there was clear hypertrophy and lamellar fusion in the gills tissues examined under microscope. Furthermore, intriguing results were obtained, such as a substantial increase in the weight gain, specific growth rate (%BW/day), FCE%, survival%, and improved secretions of amylase, cellulase, and protease activities. Likewise, high crude protein, fats content, while low moisture and total Ash was noticed in EXP I (Probiotics supplemented) compared to CTRL. The microscopic and biochemical analysis disclosed the positive role of probiotics on the fish health, thus we suggest its supplementations in the fish diet.

Duration of stunting impacts compensatory growth and carcass quality of farmed milkfish, Chanos chanos (Forsskal, 1775) under field conditions.

An 18-months field trial was performed to explore the effect of duration of stunting on growth, digestive enzymes and carcass quality in Chanos chanos. Milkfish fry (weight of 1.25 ± 0.03 g and length of 5.53 ± 0.03 cm) were stocked in earthen ponds of 0.02 ha, in triplicate, for different duration of stunting, viz., 4 months (Treatment-1; T4), 8 months (Treatment-2; T8) and 12 months (Treatment-3; T12) and a normal seed (Control; C) separately. In the stunting phase, fish were stocked at higher stocking density (0.2 million/ha) and fed de-oiled rice bran at sub-optimal level. Post-stunting or re-feeding phase commenced immediately after completion of respective stunting duration and fish were reared for the rest of the period to complete the total rearing period of 18 months. In post-stunting, fish stocking density was adjusted to (5000 pieces/ha) and fed at an optimum level (3%). At the end of stunting phase, the study found a significant reduction in growth, survival, digestive enzymes activity, except protease in the T4 group, and carcass nutrients composition of stunted fish. However, in the initial phase of post-stunting, T8 group exhibited an elevated specific growth rate (5.00 ± 0.092%/day), body weight gain (80.82 ± 1.28 g), amylase (0.585 ± 0.021 U/mg protein), protease (5.48 ± 0.13 U/mg protein), and lipase activity (7.92 ± 0.32 U/mg protein). All stunted fish groups displayed a compensatory growth response in post-stunting, but a complete growth compensation was observed in T8 group, which resulted in better feed conversion ratio (3.03 ± 0.04) feed efficiency ratio (0.33 ± 0.01), protein efficiency ratio (1.91 ± 0.03), survival (91.38 ± 0.07%) and digestive enzyme activities. Similarly, at the end of post-stunting, carcass analysis revealed a complete restoration of nutrients in stunted fish and significantly higher protein content in T8 group. Further, the study found lower meat and higher bone contents in normally reared fish than the post-stunted fish which revealed the carcass quality improvement in post-stunted fish thus indicates superiority of the stunting process over normal rearing. Overall, the study suggests that stunting of milkfish, for 8 months (T8), positively affects its growth, survival, digestive enzyme activities and carcass quality which in turn, shall help to overcome the contemporary challenges in milkfish culture.

Impacts of deepwater horizon oil on fish.

An explosion on the Deepwater Horizon (DWH) oil rig in 2010 lead to the largest marine oil spill to occur in US history, resulting in significant impacts to the ecosystems and organisms in the Northern Gulf of Mexico (GoM). The present review sought to summarize and discuss findings from the 50+ peer-reviewed publications reporting effects of DWH oil exposure on teleost fish, and concludes that oil toxicity is a multi-target, multi-organ syndrome with substantial species-specific sensitivity differences. Of the 15 species tested with characterized exposures, 20% show effects at concentrations <1 µg l-1 while 50% display effects at <8.6 µg l-1 ΣPAH50, concentrations well within the range of reported environmental levels during the spill. Cardiotoxic effects are among the most frequently reported endpoints in DWH oil exposure studies and are thought to have significant downstream effects on fitness and survival. However, additional and possibly cardio-toxic independent impacts on sensory function and behavior are reported at very low exposure concentrations (< 1 µg l-1 ∑PAH50) and are clearly deserving of further study. Available information about modes of action leading to different categories of effects are summarized in the present review. An overview of the literature illustrates that early life stages (ELS) are approximately 1-order of magnitude more sensitive than corresponding later life stages, but also illustrates that adults can be impacted at concentrations as low as 4 µg l-1 ΣPAH50. The majority of studies exploring DWH oil toxicity in fish are performed using acute exposures (1-2 days), mid-range test temperatures (26-28 °C) and measure effects at the molecular to organismal levels, leaving a pressing need for more long-term exposures, exposures at the upper and lower levels of GoM relevant temperatures, and studies investigating population level impacts.

Microbial Poly-3-Hydroxybutyrate (PHB) as a Feed Additive for Fishes and Piglets.

The large-scale use of petrochemical-based plastics is damaging our environment. Discarded plastics are harmful to both marine and land animals, sometimes causing death when ingested. Biodegradable plastics have gained attentions from the public and the academia to reduce environmental burdens. Poly-3-hydroxybutyrate (PHB), the simplest and the best-studied bioplastic member of the polyhydroxyalkanoate (PHA) family synthesized by many bacteria, has been studied as a feed additive for large yellow croaker fish and weaned piglets. The fish grow faster and gain more weight when 1% and 2% PHB is added as a feed additive, accompanied by increased survival rates. Weaned piglets are found to grow normally and showed no significant change in average daily weight gains, average daily feed intakes, feed efficiency, and organ developments when 0.5% PHB is added to the feed. It can therefore be concluded that biodegradable and biocompatible PHB is not harmful as a feed additive for marine large yellow croakers and sensitive weaned piglets. PHB therefore holds great promise as a plastic that combines biodegradability and biocompatibility with good tolerability as a feed supplement for animals.

Bioprocessed poultry by-product meals on growth, gut health and fatty acid synthesis of juvenile barramundi, Lates calcarifer (Bloch).

Poultry by-product meal (PBM) has been utilised as a substitute of fishmeal (FM) in many aquaculture species. However, little information is known regarding the use of bioprocessed PBM (BPBM) in aquaculture production. This study was undertaken to investigate whether replacing FM with BPBM improved growth performance, gut morphology and fatty acid synthesis of juvenile barramundi, Lates calcarifer. The PBM was bioprocessed by baker yeast, Saccharomyces cerevisae and Lactobacillus casei. The BPBM was used to replace FM at 75% and 100% (75BPBM and 100BPBM) contrasting against unprocessed PBM (75PBM and 100PBM) at the same levels and FM based diets as the control. Juvenile barramundi with a mean initial weight of 3.78±0.16 g were stocked at a density of 20 fish per tank. After the 42 days of study, the final weight, specific growth rate and feed conversion ratios of fish fed 75PBM and 75BPBM were not significantly different from the control. However, 100% supplementation diets of 100PBM and 100BPBM resulted in reduced performance in all growth and feed variables except total feed intake and survival. The hind gut microvillus density was significantly higher (P<0.05) in fish fed 75BPBM, whereas the microvillus diameter remained unaffected with the other experimental diets when compared to the control. A reduction in eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids of fish muscles led to a lower Σn-3/Σn-6 ratio in all dietary groups when compared to the control. The percentage of Σn-3 PUFAs decreased in 100% FM replacement diets of 100PBM and 100BPBM, while Σn-6 PUFAs increased when both bioprocessed and unprocessed PBM protein was increased in the diets. Fish fed bioprocessed diets had higher fatty acid hypocholesterolemic/hypercholesterolemic ratios (HH), indicating improved suitability for human consumption.