A review of carotenoid utilisation and function in crustacean aquaculture

Crustaceans are cultured extensively around the world in intensive farming systems. High-performance formulated feeds have been developed for crustacean aquaculture, which are often supplemented with a number of natural and synthetic carotenoid sources. Studies over a number of years have consistently shown that dietary carotenoid supplementation is beneficial for crustacean aquaculture across a range of commercially relevant parameters. Most obvious is the effect on pigmentation, where carotenoid inclusion levels in feeds and duration of feeding diets with carotenoids have been optimised across many species to improve product colour, and subsequently quality and price. However, beneficial effects of carotenoid inclusion have increasingly been demonstrated on other parameters. This review updates the recent progress in our understanding of dietary carotenoid utilisation and storage, and the combined effects of diet, genetics and environment on crustacean pigmentation. In addition, the range of other physiological benefits this class of molecules brings to these animals is summarised. These include improvements in survival, growth, reproductive capacity, disease resistance and stress resistance. © 2015 Wiley Publishing Asia Pty Ltd

An analysis of the effects of different dietary macronutrient energy sources on the growth and energy partitioning by juvenile barramundi, Lates calcarifer, reveal a preference for protein-derived energy

It is generally considered that fish respond to dietary energy densities on a consistent basis irrespective of what macronutrient source the dietary energy originates from. To test this assumption, two experiments were undertaken to establish the different roles of protein, lipid and starch as energy sources in underpinning nutritional bioenergetics in juvenile barramundi, Lates calcarifer. To do this, a range of ingredients were evaluated for their digestible protein (DP) and digestible energy (DE) value. Following this, a series of diets were formulated to an equivalent DE basis, and observed a minimum DP:DE ratio required for fish of 80 g. However, in each of the diets the proportion of DE available from protein, lipid or starch was varied to bias the contribution of each macronutrient on the origin that DE when fed to the fish. Growth of fish fed the protein diet was better than those fed the lipid diet, which was better than those fed the starch diet. Feed intake was lower in the protein diet than the lipid diet, and both were lower than the starch diet. Feed conversion was most efficient in the protein diet fed fish, which was better than the lipid diet fed fish, which was better than the starch diet fed fish. Whole fish composition varied among treatments, with differences observed in the dry matter composition, whole body lipid and gastrointestinal tract lipid content. Typically, lipid and dry matter composition were in synchrony and were usually higher in the starch fed fish and lower in the lipid fed fish. When flux of protein, lipid and energy was assessed in terms of deposition efficiencies, some significant differences were observed. Protein deposition efficiency was relatively conservative, but ranged from 33% in the starch diet fed fish to 41% in the lipid diet fed fish. Lipid deposition efficiency was more dramatic; ranging from 40% in the lipid diet to 182% in the starch diet. Energy deposition efficiency was relatively conservative among treatments, ranging from 50% to 56% efficient. Overall, the results from this study show that there is a clear hierarchy in preference for energy substrates by juvenile barramundi, such that protein > lipid > starch

The effect of marine and non-marine phospholipid rich oils when fed to juvenile barramundi (Lates calcarifer)

An experiment was conducted to assess the response of juvenile barramundi (Lates calcarifer) to four diets containing either marine- or non-marine derived neutral lipid (NL) or polar lipid (PL) sources for eight weeks in a 2 × 2 factorial design. The four diets contained 8.2% added lipid composed of a 1% fish oil base with 7.2% test lipid (n - 3 NL: Fish oil, n - 3 PL: Krill oil, n - 6 NL: Soybean oil, n - 6 PL: Soybean lecithin). The results demonstrated that the different lipid sources (either n - 3 or n - 6 omega series from either NL or PL class) had significant effects on growth performance and feed utilisation with some interaction terms noted. Growth was negatively affected in the n - 6 NL fish and the feed conversion (FCR) was highest in the n - 6 PL fish. Digestibility of total lipid and some specific fatty acids (notably 18:2n - 6 and 18:3n - 3) were also negatively affected in the n - 6 PL fish. Analysis of the whole body neutral lipid fatty acid composition showed that these mirrored those of the diets and significant interaction terms were noted. However, the whole body polar lipid fatty acids appeared to be more tightly regulated in comparison. The blood plasma biochemistry and hepatic transcription of several fatty acid metabolism genes in the n - 6 PL fed and to a lesser extent in the n - 6 NL fed fish demonstrated a pattern consistent with modified metabolic function. These results support that there are potential advantages in using phospholipid-rich oils however there are clear differences in terms of their origin. Statement of relevance: Juvenile barramundi may benefit from dietary phospholipid. © 2016 Elsevier B.V

The combined effects of diet, environment and genetics on pigmentation in the Giant Tiger Prawn, Penaeus monodon

The colour of prawns, particularly the Giant Tiger Prawn Penaeus monodon, is highly desired and fetches premium market prices. Prawn pigmentation is influenced by the interaction of a range of factors, including the amount of dietary carotenoid, the distribution of hypodermal pigments, and genetics. These aspects have been studied in isolation, but there is limited knowledge on how these components interact to influence prawn pigmentation. This study tracked the colour of prawns that had been fed four different levels of dietary astaxanthin (Axn) over 6. weeks, and then transferred to either black or white coloured tanks. The dietary influence on colour was slow and had only developed after 6. weeks. Meanwhile the effect of background colour was rapid, within 15. min. Results showed that diet and background colour work in combination to affect prawn colour. The poorest colour was recorded in prawns fed without dietary Axn and transferred to white substrates, and this colour was improved by the addition of dietary Axn. Animals fed without dietary Axn and exposed to black substrates showed an intermediate colour, and this was further improved by the addition of dietary Axn. The best colour was recorded in prawns fed 100. mg/kg Axn and exposed to black substrates. The abundance of the epithelial pigment protein crustacyanin (CRCN) was not correlated with prawn colour, suggesting that this protein does not regulate the modifications in response to background colour. Finally, the effect of substrate exposure was assessed on farmed prawns, and indicated a small positive effect on colour during harvesting. These data demonstrate that while short term exposure to black substrates can have positive effects on prawn colour, dietary Axn supplementation can both improve pigmentation of animals exposed to black substrates, and prevent the negative effects of exposure to white substrates

Dietary astaxanthin levels affect colour, growth, carotenoid digestibility and the accumulation of specific carotenoid esters in the Giant Tiger Shrimp, Penaeus monodon

The carotenoid astaxanthin (Axn) plays a vital role in shrimp pigmentation, with direct influence on product quality, and forms a significant cost component of shrimp aquaculture feeds. However, the effects of dietary Axn on other measures of shrimp physiological performance are varied, and the efficiency of carotenoid uptake from the diet and deposition in shrimp tissues is poorly defined. This study fed juvenile shrimp (Penaeus monodon) diets that contained 0, 25, 50 or 100 mg kg-1 Axn for 6 weeks. Shrimp fed carotenoid-free diets had significantly reduced colour and growth than those fed carotenoids, but survival was unaffected. Carotenoid digestibility improved as dietary carotenoid levels increased, and was 98.5% in shrimp fed 100 mg kg-1 Axn. After 6 weeks, whole body carotenoid levels were significantly depleted in 0 or 25 mg kg-1 fed shrimp, compared with those fed 50 or 100 mg kg-1 or compared with initial shrimp. This study also showed that Axn monoesters were enriched with saturated fatty acids, whereas Axn diesters were enriched with monounsaturated and polyunsaturated fatty acids. Combined, these studies demonstrate that a total dietary carotenoid intake of between 25 and 50 mg kg-1 Axn is required for normal shrimp growth and health in P. monodon. Evidence suggests that there is a functional role for the accumulation of carotenoids and the formation of specific Axn fatty acid esters, and these may be linked to the metabolism, storage, mobilization or deposition of Axn within various tissues

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

Effects of canola meal on growth, feed utilisation, plasma biochemistry, histology of digestive organs and hepatic gene expression of barramundi (Asian seabass; Lates calcarifer)

The serial replacement of fish meal (anchovetta) by canola meal (CM) (100, 200, 300 g kg−1 as either solvent extracted (SE) CM or expeller extracted (EX) CM was undertaken to investigate the effects of increasing dietary CM levels on feed intake, growth, protein and energy retention, plasma biochemistry and the expression of a suite of hepatic genes in barramundi (Asian seabass; Lates calcarifer) over an eight week feeding trial. An additional diet using lupin kernel meal (LM) to replace the fish meal was also included as a comparative reference. Eight iso-digestible nitrogenous (423±29 g kg−1) and iso-digestible energetic (14.6±8 MJ kg−1 DM) diets were formulated. Each diet was randomly allocated to triplicate groups of fish in seawater tanks (600 L), and each tank was stocked with 15 fish (53.4±7.0 g). Fish were fed once daily (9:00–10:00) to apparent satiation, and uneaten feed was collected to determine feed consumption. The results showed that the survival, feed intake, growth, FCR, energy and protein retention of fish fed the diet containing SE CM were similar or even higher to those of fish fed the fish meal reference diet (FM) and the LM diet. Fish fed with the diet containing 300 g kg−1 SE CM did not show any changes in biochemistry and gene expression in a suite of detoxification genes. However, the diet with 300g kg−1 EX CM depressed feed intake, growth performance and increased feed conversion ratio (FCR). Transcription of genes involving in fatty acid synthesis and the TCA cycle were not changed by different diets. The down regulation of gene expression in certain detoxification genes (Lc CYP1A1, Lc CYP3A, Lc CYP2N and Lc GST) was observed in fish fed with the diet containing 300 g kg−1 EX CM compared to the FM control diet and other experimental diets. In general, the SE CM can be used up to 300 g kg−1 diet without negative performance effects or signs of clinical plasma biochemistry. By contrast the maximum acceptable level of the EX CM for barramundi was only 200 g kg−1. Higher inclusion level of the EX CM induced negative effects on growth performance, feed utilisation, plasma biochemistry and gene expression in relation to detoxification

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

An evaluation of the complete replacement of both fishmeal and fish oil in diets for juvenile Asian seabass, Lates calcarifer

An experiment was conducted to examine the potential for the complete replacement of fishmeal (FM) and fish oil (FO) in diets for barramundi,Lates calcarifer. A series of diets were formulated to the same digestible protein and energy specifications, but which were designed with FM inclusion levels at 300, 200, 100 or 0g/kg and FO at 100%, 30%, 15% or 0% of the added oil in the diets (4×4 factorial design). Ricebran oil was the alternative oil used in the growth study, while soybean meal and poultry meal were the main alternative protein sources used. For the growth study, fish of an initial weight of 154.4±1.1g were randomly allocated across 48 tanks (three replicates per treatment). After eight weeks, the average weight gain across all treatments was 187.7±2.3g/fish and feed conversion across all treatments averaged 1.04±0.01 feed/gain. A significant effect of FM on both feed intake and weight gain was observed, and this was observed as early as within the first few weeks, but no similar such effect was observed with FO. No effects were observed on protein deposition efficiency, though both lipid and energy deposition efficiencies were affected by FM level. The reduction in FO had a notable effect on the fatty acid composition of the diets and subsequently the fish fatty acid composition. Expression of key LC-PUFA metabolism genes in the liver of the fish was influenced by both FM and FO levels, but was only significant at the extremes of the treatment ranges. The results from this study demonstrate that there is clear potential to replace almost all the FM content of barramundi diets without loss of fish performance, up to and including diets with as little as 100g/kg fishmeal. Replacement of fish oil was more successful with the ability to completely replace all FO demonstrated at all but the lowest inclusion levels of FM. These results clearly demonstrate that the near complete replacement of both FM and FO in barramundi diets is a technical reality

An hypothesis on crustacean pigmentation metabolism:L-carnitine and nuclear hormone receptors as limiting factors

Astaxanthin (Axn) is the primary pigment molecule in crustaceans associated with quality, health and growth traits, leading to increased marketing value. Axn can be contained within the protein complex crustacyanin (CRCN) to produce an array of different shell colours, or esterified with fatty acids (FA) for storage but also contributing additional red colouration. l-Carnitine (LC) has a major role in FA oxidation and mitochondrial function optimization, which could influence the proportion of Axn complexed with FA or CRCN. Peroxisome proliferator activated receptors (PPARs) have important roles in FA and Axn uptake, and stored lipid oxidation affecting Axn homeostasis and storage in lipid bodies. Whether Axn could increase PPAR signalling and carnitine palmitoyl transferase activity, leading to induction of lipid metabolism, is not known in crustaceans. Several FA have been shown to preferentially form FA Axn-esters, including saturated fatty acids (SFA) such as C16:0 and C18:0, mono-unsaturated fatty acids (MUFA) such as C16:1 and C18:1, and poly-unsaturated fatty acids (PUFA) such as C20:4, C20:5, and C20:6. We hypothesize that manipulating the dietary ratios and inclusion of LC, Axn, and specific FA may be able to further improve pigment utilization, lipid metabolism, health, and growth in crustaceans.</p