Adequate supply of dietary taurine stimulates expression of molecular markers of growth and protein turnover in juvenile barramundi (Lates calcarifer).

A trial was conducted to investigate the effect of dietary taurine (Tau) supply on the plasma amino acid composition and hepatic expression of several genes in juvenile barramundi (Lates calcarifer) after feeding. Triplicate tanks of fish (average weight, 89.3 g) were fed diets containing either a deficient (1 g kg-1), adequate (8 g kg-1) or excessive (19 g kg-1) level of dietary Tau. Liver tissues collected before feeding, and at 2- and 4-h post-feeding, were analysed for expression of genes involved in pathways of sulphur amino acid turnover, Tau biosynthesis and transport, target of rapamycin (TOR) signalling, the somatotropic axis and protein turnover. The treatment had no significant effect on the profiles of any amino acid in plasma collected over time after feeding, other than Tau and glycine. The expression profile of cystine and Tau synthetic genes suggested an effect of Tau excess on the metabolism of cystine. Markers of two pathways of Tau biosynthesis appear to be active in this species, providing proof that this species possesses the ability to synthesise Tau from SAA precursors. A marker for the regulation of Tau transport and homeostasis was shown to be directly regulated by Tau availability, whilst a link between adequate supply of Tau and TOR pathway-mediated growth stimulation was also apparent. An observed depression in expression of genes of the somatotropic axis, coupled with upregulation of the proteolytic and TOR-suppressing genes, in response to excessive Tau supply in the diet, signalled that excessive Tau may not be conducive to optimal growth in this species.

Endogenous biosynthesis of n-3 long-chain PUFA in Atlantic salmon.

A more efficient utilisation of marine-derived sources of dietary n-3 long-chain PUFA (n-3 LC PUFA) in cultured Atlantic salmon (Salmo salar L.) could be achieved by nutritional strategies that maximise endogenous n-3 LC PUFA synthesis. The objective of the present study was to quantify the extent of n-3 LC PUFA biosynthesis and the resultant effect on fillet nutritional quality in large fish. Four diets were manufactured, providing altered levels of dietary n-3 substrate, namely, 18 : 3n-3, and end products, namely, 20 : 5n-3 and 22 : 6n-3. After 283 d of feeding, fish grew in excess of 3000 g and no differences in growth performance or biometrical parameters were recorded. An analysis of fatty acid composition and in vivo metabolism revealed that endogenous production of n-3 LC PUFA in fish fed a diet containing no added fish oil resulted in fillet levels of n-3 LC PUFA comparable with fish fed a diet with added fish oil. However, this result was not consistent among all treatments. Another major finding of this study was the presence of abundant dietary n-3 substrate, with the addition of dietary n-3 end product (i.e. fish oil) served to increase final fillet levels of n-3 LC PUFA. Specifically, preferential ß-oxidation of dietary C18 n-3 PUFA resulted in conservation of n-3 LC PUFA from catabolism. Ultimately, this study highlights the potential for endogenous synthesis of n-3 LC PUFA to, partially, support a substantial reduction in the amount of dietary fish oil in diets for Atlantic salmon reared in seawater.

Endogenous production of n-3 long-chain PUFA from first feeding and the influence of dietary linoleic acid and the α-linolenic:linoleic ratio in Atlantic salmon (Salmo salar).

Atlantic salmon (Salmo salar) possess enzymes required for the endogenous biosynthesis of n-3 long-chain PUFA (LC-PUFA), EPA and DHA, from α-linolenic acid (ALA). Linoleic acid (LA) competes with ALA for LC-PUFA biosynthesis enzymes leading to the production of n-6 LC-PUFA, including arachidonic acid (ARA). We aimed to quantify the endogenous production of EPA and DHA from ALA in salmon fed from first feeding on diets that contain no EPA and DHA and to determine the influence of dietary LA and ALA:LA ratio on LC-PUFA production. Salmon were fed from first feeding for 22 weeks with three diets formulated with linseed and sunflower oils to provide ALA:LA ratios of approximately 3:1, 1:1 and 1:3. Endogenous production of n-3 LC-PUFA was 5·9, 4·4 and 2·8 mg per g fish and that of n-6 LC-PUFA was 0·2, 0·5 and 1·4 mg per g fish in salmon fed diets with ALA:LA ratios of 3:1, 1:1 and 1:3, respectively. The ratio of n-3:n-6 LC-PUFA production decreased from 27·4 to 2·0, and DHA:EPA ratio increased and EPA:ARA and DHA:ARA ratios decreased, as dietary ALA:LA ratio decreased. In conclusion, with a dietary ALA:LA ratio of 1, salmon fry/parr produced about 28 µg n-3 LC-PUFA per g fish per d, with a DHA:EPA ratio of 3·4. Production of n-3 LC-PUFA exceeded that of n-6 LC-PUFA by almost 9-fold. Reducing the dietary ALA:LA ratio reduced n-3 LC-PUFA production and EPA:ARA and DHA:ARA ratios but increased n-6 LC-PUFA production and DHA:EPA ratio.

Coping with climatic extremes: Dietary fat content decreased the thermal resilience of barramundi (Lates calcarifer).

Aquatic organisms, including important cultured species, are forced to contend with acute changes in water temperature as the frequency and intensity of extreme weather events worsen. Acute temperature spikes are likely to threaten aquaculture species, but dietary intervention may play an important protective role. Increasing the concentration of macronutrients, for example dietary fat content, may improve the thermal resilience of aquaculture species, however, this remains unexplored. To evaluate this hypothesis, we used two commercially available diets (20% versus 10% crude fat) to examine if dietary fat content improves the growth performance of juvenile barramundi (Lates calcarifer) while increasing their resilience to acute thermal stress. Fish were fed their assigned diets for 28-days before assessing the upper thermal tolerance (CTMAX) and the thermal sensitivity of swimming performance (UCRIT) and metabolism. We found that feeding fish a high fat diet resulted in heavier fish, but did not affect the thermal sensitivity of swimming performance or metabolism over an 18 °C temperature range (from 20 to 38 °C). Thermal tolerance was compromised in fish fed the high fat diet by 0.48 °C, showing significantly lower CTMAX. Together, these results suggest that while a high fat diet increases juvenile L. calcarifer growth, it does not benefit physiological performance across a range of relevant water temperatures and may even reduce fish tolerance of extreme water temperatures. These data may have implications for aquaculture production in a warming world, where episodic extremes of temperature are likely to become more frequent.

Impact of dietary starch on extrahepatic tissue lipid metabolism in farmed European (Dicentrarchus labrax) and Asian seabass (Lates calcarifer).

In aquaculture, there is high interest in substituting marine-derived with vegetable-based ingredients as energy source. Farmed carnivorous fish under high carbohydrate diets tend to increase adiposity but it remains unclear if this happens by increased lipid retention/accumulation, promotion of lipogenic pathways, or both. In order to determine the response of extrahepatic tissue to dietary starch, European (Dicentrarchus labrax) and Asian (Lates calcarifer) seabass were fed a control (low starch; LS) or experimental (high starch; HS) diet, for at least 21 days and then transferred for 6 days to saltwater enriched with deuterated water 2H2O. Incorporation of 2H-labelling follows well-defined metabolic steps, and analysis of triacylglycerols (TAG) 2H-enrichment by 2HNMR allowed evaluation of de novo lipogenesis (DNL) in muscle and visceral adipose tissue (VAT). Fractional synthetic rates for TAG-bound fatty acids and glycerol were quantified separately providing a detailed lipogenic profile. The FA profile differed substantially between muscle and VAT in both species, but their lipogenic fluxes revealed even greater differences. In European seabass, HS promoted DNL of TAG-bound FA, in muscle and VAT. High 2H-enrichment also found in muscle TAG-bound glycerol was indicative of its role on lipid cycling. In Asian seabass, HS had no effect on muscle FA composition and lipogenic flux, with no 2H-enriched TAG being detected. VAT on the other hand revealed a strong enhancement of DNL in HS-fed fish along with high TAG-bound glycerol cycling. This study consolidated the use of 2H2O as tracer for fish lipid metabolism in different tissues, under different dietary conditions and suitable to use in different fish models.

An analysis of partial efficiencies of energy utilisation of different macronutrients by barramundi (Lates calcarifer) shows that starch restricts protein utilisation in carnivorous fish.

This study examined the effect of including different dietary proportions of starch, protein and lipid, in diets balanced for digestible energy, on the utilisation efficiencies of dietary energy by barramundi (Lates calcarifer). Each diet was fed at one of three ration levels (satiety, 80 % of initial satiety and 60 % of initial satiety) for a 42-d period. Fish performance measures (weight gain, feed intake and feed conversion ratio) were all affected by dietary energy source. The efficiency of energy utilisation was significantly reduced in fish fed the starch diet relative to the other diets, but there were no significant effects between the other macronutrients. This reduction in efficiency of utilisation was derived from a multifactorial change in both protein and lipid utilisation. The rate of protein utilisation deteriorated as the amount of starch included in the diet increased. Lipid utilisation was most dramatically affected by inclusion levels of lipid in the diet, with diets low in lipid producing component lipid utilisation rates well above 1·3, which indicates substantial lipid synthesis from other energy sources. However, the energetic cost of lipid gain was as low as 0·65 kJ per kJ of lipid deposited, indicating that barramundi very efficiently store energy in the form of lipid, particularly from dietary starch energy. This study defines how the utilisation efficiency of dietary digestible energy by barramundi is influenced by the macronutrient source providing that energy, and that the inclusion of starch causes problems with protein utilisation in this species.

Atlantic Salmon (Salmo salar L.) Gastrointestinal Microbial Community Dynamics in Relation to Digesta Properties and Diet.

To better understand salmon GI tract microbial community dynamics in relation to diet, a feeding trial was performed utilising diets with different proportions of fish meal, protein, lipid and energy levels. Salmon gut dysfunction has been associated with the occurrence of casts, or an empty hind gut. A categorical scoring system describing expressed digesta consistency was evaluated in relation to GI tract community structure. Faster growing fish generally had lower faecal scores while the diet cohorts showed minor differences in faecal score though the overall lowest scores were observed with a low protein, low energy diet. The GI tract bacterial communities were highly dynamic over time with the low protein, low energy diet associated with the most divergent community structure. This included transiently increased abundance of anaerobic (Bacteroidia and Clostridia) during January and February, and facultatively anaerobic (lactic acid bacteria) taxa from February onwards. The digesta had enriched populations of these groups in relation to faecal cast samples. The majority of samples (60-86 %) across all diet cohorts were eventually dominated by the genus Aliivibrio. The results suggest that an interaction between time of sampling and diet is most strongly related to community structure. Digesta categorization revealed microbes involved with metabolism of diet components change progressively over time and could be a useful system to assess feeding responses.

Defining the allometric relationship between size and individual fatty acid turnover in barramundi Lates calcarifer.

An experiment was conducted with barramundi (Asian seabass; Lates calcarifer) to examine the allometric scaling effect of individual fatty acids. Six treatment size classes of fish were deprived of food for 21days (Treatment A, 10.5±0.13g; Treatment B, 19.2±0.11g; Treatment C, 28.3±0.05g; Treatment D, 122.4±0.10g; Treatment E, 217.6±0.36g; Treatment F, 443.7±1.48g; mean±SD) with each treatment comprising of fifteen fish, in triplicate. The assessment of somatic losses of whole-body energy and lipid were consistent with previous studies, validating the methodology to be extended to individual fatty acids. Live-weight (LW) exponent values were determined to be 0.817±0.010 for energy and 0.895±0.007 for lipid. There were significant differences among the fatty acids ranging from 0.687±0.005 for 20:5n-3 (eicosapentaenoic acid) and 0.954±0.008 for 18:1n-9 (oleic acid). The LW exponent values were applied to existing fatty acid intake and deposition data of barramundi fed with either 100% fish oil or 100% poultry oil. From this the maintenance requirement for each fatty acid was determined. The metabolic demands for maintenance and growth were then iteratively determined for fish over a range of size classes. Application of these exponent values to varying levels of fatty acid intake demonstrated that the biggest driver in the utilisation of fatty acids in this species is deposition demand and despite their reputed importance, the long-chain polyunsaturated fatty acids had nominal to no maintenance requirement.

A comparative analysis of the response of the hepatic transcriptome to dietary docosahexaenoic acid in Atlantic salmon (Salmo salar) post-smolts.

BACKGROUND: The present study aimed to explore the impact of dietary docosahexaenoic acid (DHA) on aspects of the metabolism of Atlantic salmon (Salmo salar). The effects of diets containing increasing levels of DHA (1 g kg(-1), 3 g kg(-1), 6 g kg(-1), 10 g kg(-1) and 13 g kg(-1)) on the liver transcriptome of post-smolt salmon was examined to elucidate patterns of gene expression and responses of specific metabolic pathways. Total RNA was isolated from the liver of individual fish and analyzed using a custom gene expression 44K feature Atlantic salmon oligo-microarray. RESULTS: The expression of up to 911 unique annotated genes was significantly affected by dietary DHA inclusion relative to a low DHA reference diet. Analysis of a total of 797 unique genes were found with a significant linear correlation between expression level and dietary DHA. Gene-Set Enrichment Analysis (GSEA) identified a range of pathways that were significantly affected by dietary DHA content. CONCLUSIONS: Pathways that showed a significant response to dietary DHA level included those for long-chain polyunsaturated fatty acid biosynthesis, fatty acid elongation, steroid biosynthesis, glycan biosynthesis, protein export and protein processing in the endoplasmic reticulum. These findings suggest that in addition to clear roles in influencing lipid metabolic pathways, DHA might also have key functional roles in other pathways distinct from lipid metabolism.

Rapid effects of essential fatty acid deficiency on growth and development parameters and transcription of key fatty acid metabolism genes in juvenile barramundi (Lates calcarifer).

Barramundi (Lates calcarifer), a catadromous teleost of significant and growing commercial importance, are reported to have limited fatty acid bioconversion capability and therefore require preformed long-chain PUFA (LC-PUFA) as dietary essential fatty acid (EFA). In this study, the response of juvenile barramundi (47·0 g/fish initial weight) fed isolipidic and isoenergetic diets with 8·2% added oil was tested. The experimental test diets were either devoid of fish oil (FO), and thus with no n-3 LC-PUFA (FO FREE diet), or with a low inclusion of FO (FO LOW diet). These were compared against a control diet containing only FO (FO CTRL diet) as the added lipid source, over an 8-week period. Interim samples and measurements were taken fortnightly during the trial in order to define the aetiology of the onset and progression of EFA deficiency. After 2 weeks, the fish fed the FO FREE and FO LOW diets had significantly lower live-weights, and after 8 weeks significant differences were detected for all performance parameters. The fish fed the FO FREE diet also had a significantly higher incidence of external abnormalities. The transcription of several genes involved in fatty acid metabolism was affected after 2 weeks of feeding, showing a rapid nutritional regulation. This experiment documents the aetiology of the onset and the progression of EFA deficiency in juvenile barramundi and demonstrates that such deficiencies can be detected within 2 weeks in juvenile fish.

Interactions between dietary docosahexaenoic acid and other long-chain polyunsaturated fatty acids on performance and fatty acid retention in post-smolt Atlantic salmon (Salmo salar).

A study with varying dietary inclusion levels (1, 5, 10, 15 and 20 g kg(-1)) of docosahexaenoic acid (DHA; 22:6n-3) was conducted with post-smolt (111 ± 2.6 g; mean ± S.) Atlantic salmon (Salmo salar) over a 9-week period. In addition to the series of DHA inclusion levels, the study included further diets that had DHA at 10 g kg(-1) in combination with either eicosapentaenoic acid (EPA; 20:5n-3) or arachidonic acid (ARA; 20:4n-6), both also included at 10 g kg(-1). An additional treatment with both EPA and DHA included at 5 g kg(-1) (total of 10 g kg(-1) long-chain polyunsaturated fatty acids, LC-PUFA) was also included. After a 9-week feeding period, fish were weighed, and carcass, blood and tissue samples collected. A minor improvement in growth was seen with increasing inclusion of DHA. However, the addition of EPA further improved growth response while addition of ARA had no effect on growth. As with most lipid studies, the fatty acid composition of the whole body lipids generally reflected that of the diets. However, there were notable exceptions to this, and these implicate some interactions among the different LC-PUFA in terms of the fatty acid biochemistry in this species. At very low inclusion levels, DHA retention was substantially higher (~250%) than that at all other inclusion levels (31-58%). The inclusion of EPA in the diet also had a positive effect on the retention efficiency of DHA. However, EPA retention was highly variable and at low DHA inclusion levels there was a net loss of EPA as this fatty acid was most likely elongated to produce DHA, consistent with increased DHA retention with additional EPA in the diet. Retention of DPA (22:5n-3) was high at low levels of DHA, but diminished with increasing DHA inclusion, similar to that seen with DHA retention. The addition of EPA to the diet resulted in a substantial increase in the efficiency of DPA retention; the inclusion of ARA had the opposite effect. Retention of ARA was unaffected by DHA inclusion, but the addition of either EPA or ARA to the diet resulted in a substantial reduction in the efficiency of ARA retention. No effects of dietary treatment were noted on the retention of either linolenic (18:3n-3) or linoleic (18:2n-6) acids. When the total n-3 LC-PUFA content of the diet was the same but consisted of either DHA alone or as a combination of EPA plus DHA, the performance effects were similar.

Postprandial molecular responses in the liver of the barramundi, Lates calcarifer.

The regulation of gene expression by nutrients is an important mechanism governing energy storage and growth in most animals, including fish. At present, very few genes that regulate intermediary metabolism have been identified in barramundi, nor is there any understanding of their nutritional regulation. In this study, a partial barramundi liver transcriptome was assembled from next-generation sequencing data and published barramundi EST sequences. A large number of putative metabolism genes were identified in barramundi, and the changes in the expression of 24 key metabolic regulators of nutritional pathways were investigated in barramundi liver over a time series immediately after a meal of a nutritionally optimised diet for this species. Plasma glucose and free amino acid levels showed a mild postprandial elevation which peaked 2 h after feeding, and had returned to basal levels within 4 or 8 h, respectively. Significant activation or repression of metabolic nuclear receptor regulator genes were observed, in combination with activation of glycolytic and lipogenic pathways, repression of the final step of gluconeogenesis and activation of the Akt-mTOR pathway. Strong correlations were identified between a number of different metabolic genes, and the coordinated co-regulation of these genes may underlie the ability of this fish to utilise dietary nutrients. Overall, these data clearly demonstrate a number of unique postprandial responses in barramundi compared with other fish species and provide a critical step in defining the response to different dietary nutrient sources.

Nutrient and Energy Apparent Digestibility of Protein-Based Feed Ingredients and Effect of the Dietary Factors on Growth Performance and Feed Utilization of Sobaity Seabream, Sparidentex hasta.

Two separate feeding trials were undertaken to benchmark a series of commercial diets and determine the nutrient and energy apparent digestibility coefficients of a variety of protein-based feed ingredients when fed to sobaity seabream, Sparidentex hasta. In Experiment 1, triplicate groups of fish (initial body weight: 330.5 ± 2.6 g) were fed with one of three locally available diets containing crude protein (CP) levels ranging from 44 to 46% of dry matter (DM), each with ~12% crude fat. Fish grew at around 3.2 g day-1 with a specific growth rate (SGR) of 0.7% day-1. Both the feed conversion ratio (FCR) and protein efficiency ratio (PER) were significantly better in fish fed diets, which contained the highest (46.4%) crude protein level. Overall, the data from these preliminary studies suggest that the best performance by sobaity seabream was obtained with a diet containing 46% crude protein, 20 MJ/kg, and a protein-to-energy ratio of 23 mg/kJ. In Experiment 2, fish with an initial body weight of 319 ± 7 g were held in 11 tanks and fed reference (D1) and test diets (D2-D11) for 7 days before fecal collection. This process was repeated twice in a blocking arrangement to generate three replicates. Each of the ten test diets contained 30% of a test ingredient, with the remaining 70% proportionally identical to the D1 diet. Diet apparent digestibility coefficients (ADCs) were measured, and the diet ADCs were then used to derive the protein and energy ADCs for the individual test ingredients. Ingredient protein ADC ranged between 75.5 and 93.9%, while ingredient energy ADC ranged between 66.8 and 81.2%.

Mechanisms of colour adaptation in the prawn Penaeus monodon.

Exposure of prawns to dark- or light-coloured substrates is known to trigger a strong colour adaptation response through expansion or contraction of the colouration structures in the prawn hypodermis. Despite the difference in colour triggered by this adaptive response, total levels of the predominant carotenoid pigment, astaxanthin, are not modified, suggesting that another mechanism is regulating this phenomenon. Astaxanthin binds to a specific protein called crustacyanin (CRCN), and it is the interaction between the quantities of each of these compounds that produces the diverse range of colours seen in crustacean shells. In this study, we investigated the protein changes and genetic regulatory processes that occur in prawn hypodermal tissues during adaptation to black or white substrates. The amount of free astaxanthin was higher in animals adapted to dark substrate compared with those adapted to light substrate, and this difference was matched by a strong elevation of CRCN protein. However, there was no difference in the expression of CRCN genes either across the moult cycle or in response to background substrate colour. These results indicate that exposure to a dark-coloured substrate causes an accumulation of CRCN protein, bound with free astaxanthin, in the prawn hypodermis without modification of CRCN gene expression. On light-coloured substrates, levels of CRCN protein in the hypodermis are reduced, but the carotenoid is retained, undispersed in the hypodermal tissue, in an esterified form. Therefore, the abundance of CRCN protein affects the distribution of pigment in prawn hypodermal tissues, and is a crucial regulator of the colour adaptation response in prawns.

Effect of Lemna minor supplemented diets on growth, digestive physiology and expression of fatty acids biosynthesis genes of Cyprinus carpio.

The potential nutritional value of duckweed Lemna minor (Lemnaceae) was evaluated for common carp Cyprinus carpio fry. Fish were fed diets containing five graded levels of duckweed: 0% (LM0, control), 5% (LM5), 10% (LM10), 15% (LM15) and 20% (LM20). The final weight and specific growth rate were significantly higher in LM15 and LM20 diets fed fish compared to others. Feed conversion ratio was minimum in fish fed diet LM20. Amylase activity was significantly higher in LM0 treatment. Total protease, trypsin and chymotrypsin activities showed linear relationships with the increased level of duckweed in the diet. Protein and essential amino acids contents were significantly higher in carp fed diets LM15 and LM20 compared to others. Lipid content was significantly higher in fish fed duckweed-based diets compared to control. A direct relationship was found between the inclusion level of duckweed in the diet and n-3 long-chain polyunsaturated fatty acid (LC-PUFA) content of carp. Contents of desaturated and elongated products of dietary linolenic acid (18:3n-3) including 20:4n-3, 20:5n-3, 22:5n-3 and 22:6n-3 increased in a graded manner with increasing dietary duckweed. The monounsaturated fatty acids and n-6 PUFA contents reduced significantly in fish fed duckweed. Expression of fads2d6, elovl2, elovl5 and fas were higher in carp fed diets LM10, LM15 and LM20 compared to control fish. The inclusion of L. minor in diet enhanced the nutritional value of carp by increasing protein, lipid, amino acids and n-3 PUFA contents.

Growing backwards: an inverted role for the shrimp ortholog of vertebrate myostatin and GDF11.

Myostatin (MSTN) and growth differentiation factor-11 (GDF11) are closely related proteins involved in muscle cell growth and differentiation as well as neurogenesis of vertebrates. Both MSTN and GDF11 negatively regulate their functions. Invertebrates possess a single ortholog of the MSTN/GDF11 family. In order to understand the role of MSTN/GDF11 in crustaceans, the gene ortholog was identified and characterized in the penaeid shrimp Penaeus monodon. The overall protein sequence and specific functional sites were highly conserved with other members of the MSTN/GDF11 family. Gene transcripts of pmMstn/Gdf11, assessed by real-time PCR, were detected in a variety of tissue types and were actively regulated in muscle across the moult cycle. To assess phenotypic function in shrimp, pmMstn/Gdf11 gene expression was downregulated by tail-muscle injection of sequence-specific double-stranded RNA. Shrimp with reduced levels of pmMstn/Gdf11 transcripts displayed a dramatic slowing in growth rate compared with control groups. Findings from this study place the MSTN/GDF11 gene at the centre of growth regulation in shrimp, but suggest that, compared with higher vertebrates, this gene has an opposite role in invertebrates such as shrimp, where levels of gene expression may positively regulate growth.

Limitations to Starch Utilization in Barramundi (Lates calcarifer) as Revealed by NMR-Based Metabolomics.

Practical diets for commercial barramundi production rarely contain greater than 10% starch, used mainly as a binding agent during extrusion. Alternative ingredients such as digestible starch have shown some capacity to spare dietary protein catabolism to generate glucose. In the present study, a carnivorous fish species, the Asian seabass (Lates calcarifer) was subjected to two diets with the same digestible energy: Protein (P) - with high protein content (no digestible starch); and Starch (S) - with high digestible (pregelatinized) starch content. The effects of a high starch content diet on hepatic glycogen synthesis as well as the muscle and liver metabolome were studied using a complementary approach of 1H and 2H NMR. The hepatosomatic index was lower for fish fed high starch content diet while the concentration of hepatic glycogen was similar between groups. However, increased glycogen synthesis via the direct pathway was observed in the fish fed high starch content diet which is indicative of increased carbohydrate utilization. Multivariate analysis also showed differences between groups in the metabolome of both tissues. Univariate analysis revealed more variations in liver than in muscle of fish fed high starch content diet. Variations in metabolome were generally in agreement with the increase in the glycogen synthesis through direct pathway, however, this metabolic shift seemed to be insufficient to keep the growth rate as ensured by the diet with high protein content. Although liver glycogen does not make up a substantial quantity of total stored dietary energy in carnivorous fish, it is a key regulatory intermediate in dietary energy utilization.

In vitro characteristics of an Atlantic salmon (Salmo salar L.) hind gut microbial community in relation to different dietary treatments.

In this study, microbial community dynamics were assessed within a simple in vitro model system in order to understand those changes influenced by diet. The abundance and diversity of bacteria were monitored within different treatment slurries inoculated with salmon faecal samples in order to mimic the effects of dietary variables. A total of five complete diets and two ingredients (plant meal) were tested. The total viable counts (TVCs) and sequencing data revealed that there was very clear separation between the complete diets and the plant meal treatments, suggesting a dynamic response by the allochthonous bacteria to the treatments. Automated ribosomal intergenic spacer analysis (ARISA) results showed that different diet formulations produced different patterns of fragments, with no separation between the complete diets. However, plant-based protein ingredients were clearly separated from the other treatments. 16S rRNA Illumina-based sequencing analysis showed that members of the genera Aliivibrio, Vibrio and Photobacterium became predominant for all complete diets treatments. The plant-based protein ingredient treatments only sustained weak growth of the genus Sphingomonas. In vitro based testing of diets could be a useful strategy to determine the potential impact of either complete feeds or ingredients on major fish gastrointestinal tract microbiome members.

Eicosapentaenoic Acid, Arachidonic Acid and Eicosanoid Metabolism in Juvenile Barramundi Lates calcarifer.

A two part experiment was conducted to assess the response of barramundi (Lates calcarifer; initial weight = 10.3 ± 0.03 g; mean ± S.D.) fed one of five diets with varying eicosapentaenoic acid (diets 1, 5, 10, 15 and 20 g/kg) or one of four diets with varying arachidonic acid (1, 6, 12, 18 g/kg) against a fish oil control diet. After 6 weeks of feeding, the addition of EPA or ARA did not impact on growth performance or feed utilisation. Analysis of the whole body fatty acids showed that these reflected those of the diets. The ARA retention demonstrated an inversely related curvilinear response to either EPA or ARA. The calculated marginal utilisation efficiencies of EPA and ARA were high (62.1 and 91.9 % respectively) and a dietary ARA requirement was defined (0.012 g/kg(0.796)/day). The partial cDNA sequences of genes regulating eicosanoid biosynthesis were identified in barramundi tissues, namely cyclooxygenase 1 (Lc COX1a, Lc COX1b), cyclooxygenase 2 (Lc COX2) and lipoxygenase (Lc ALOX-5). Both Lc COX2 and Lc ALOX-5 expression in the liver tissue were elevated in response to increasing dietary ARA, meanwhile expression levels of Lc COX2 and the mitochondrial fatty acid oxidation gene carnitine palmitoyltransferase 1 (Lc CPT1a) were elevated in the kidney. A low level of EPA increased the expression of Lc COX1b in the liver. Consideration should be given to the EPA to ARA balance for juvenile barramundi in light of nutritionally inducible nature of the cyclooxygenase and lipoxygenase enzymes.

Identification of genes involved in reproduction and lipid pathway metabolism in wild and domesticated shrimps.

The aims of this study were to identify genes involved in reproduction and lipid pathway metabolism in Penaeus monodon and correlate their expression with reproductive performance. Samples of the hepatopancreas and ovaries were obtained from a previous study of the reproductive performance of wild and domesticated P. monodon broodstock. Total mRNA from the domesticated broodstock was used to create two next generation sequencing cDNA libraries enabling the identification of 11 orthologs of key genes in reproductive and nutritional metabolic pathways in P. monodon. These were identified from the library of de novo assembled contigs, including the description of 6 newly identified genes. Quantitative RT-PCR of these genes in the hepatopancreas prior to spawning showed that the domesticated mature females significantly showed higher expression of the Pm Elovl4, Pm COX and Pm SUMO genes. The ovaries of domesticated females had a significantly decreased expression of the Pm Elovl4 genes. In the ovaries of newly spawned females, a significant correlation was observed between hepatosomatic index and the expression of Pm FABP and also between total lipid content and the expression of Pm CYP4. Although not significant, the highest levels of correlation were found between relative fecundity and Pm CRP and Pm CYP4 expression, and between hatching rate and Pm Nvd and Pm RXR expression. This study reports the discovery of genes involved in lipid synthesis, steroid biosynthesis and reproduction in P. monodon. These results indicate that genes encoding enzymes involved in lipid metabolism pathways might be potential biomarkers to assess reproductive performance.