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Interest in sea urchin roe enhancement aquaculture is growing due to an increased global demand for high-quality roe that is suitable for export to international markets. Yet, fine-tuning of efficient collection methods and improved growing techniques are still key bottlenecks to industry success. Urchins suitable for roe enhancement are generally collected from rocky barrens where they occur in high densities. Few studies have investigated the efficiency of methods used to collect urchins from barrens or the associated short- and long-term handling effects due to collection methods. Here, we tested 2 methods to collect the purple sea urchin Heliocidaris erythrogramma, a species that is highly abundant in barrens in temperate waters and which is a viable candidate for roe enhancement aquaculture. We assessed short- and long-term survival, external urchin condition and final gonad indices after 12 wk of roe enhancement. Divers using a 3-pronged hook and catch bag collected urchins 1.9 times faster (392 urchins h-1) than careful hand collection (207 urchins h-1). Collection method did not significantly influence mortality rate, external health or gonad indices after 12 wk of roe enhancement. Our results show that H. erythrogramma in barrens is robust to rapid mechanical collection by divers, which increases its suitability as a candidate capture-based aquaculture species.

Overgrazing by the overabundant native purple urchin Heliocidaris erythrogramma has caused kelp-dominated reefs to shift to urchin barrens throughout southeastern Australia. These areas are characterised by low kelp abundance, low biodiversity and high urchin densities. As purple urchin gonads are a delicacy in many countries, commercial harvest from barrens could aid kelp recovery. However, the lack of macroalgae in these habitats, driven by high urchin densities, results in urchins with small, poor-quality roe that is commercially undesirable. To overcome this, we assessed whether urchin gonad quantity and quality could be improved with access to high-quality feed and optimal environmental conditions, a process known as ‘gonad conditioning’. Specifically, we (1) surveyed the quality of urchins from barrens and kelp sites in Port Phillip Bay, Australia, over 18 mo and (2) tested if gonad conditioning was effective on urchins from barrens during and after the harvest season. Field surveys revealed considerable variation in gonad size across sites, habitats and collection periods (mean gonad index range: 3 to 12%). Gonad conditioning with the best diet increased urchin gonad size by up to 2.8 times during the harvest season. Moreover, gonads of conditioned urchins from one barren were 3 times brighter in colour and contained lower concentrations of arsenic than wild urchins. In contrast, gonad conditioning at 22°C after the harvest season was ineffective. Our results show that targeted in-season harvest from barrens and subsequent gonad conditioning produces roe of commercial quality, promoting the use of urchin fisheries as a tool for managing urchin barrens.

The mucosal surfaces of fish, including their intestines, gills, and skin, are constantly exposed to various environmental threats, such as water quality fluctuations, pollutants, and pathogens. However, various cells and microbiota closely associated with these surfaces work in tandem to create a functional protective barrier against these conditions. Recent research has shown that incorporating specific feed ingredients into fish diets can significantly boost their mucosal and general immune response. Among the various ingredients being investigated, insect meal has emerged as one of the most promising options, owing to its high protein content and immunomodulatory properties. By positively influencing the structure and function of mucosal surfaces, insect meal (IM) has the potential to enhance the overall immune status of fish. This review provides a comprehensive overview of the potential benefits of incorporating IM into aquafeed as a feed ingredient for augmenting the mucosal immune response of fish., Competing Interests: Declaration of competing interest The authors have no person relationship or financial interests that could appeared to the influence of work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Determining the macronutrient requirements for commercially valuable aquaculture species remains crucial for maximising production efficiency. Yet, such information is lacking for Australian hybrid abalone ( Haliotis rubra × Haliotis laevigata ), particularly with respect to life stage and water temperatures. The present study aimed to evaluate the effect of dietary protein inclusion level on the growth performance, nutrient utilisation and nutritional quality of juvenile (3·3 g) Australian hybrid abalone reared at three different temperatures representative of winter (12°C), average annual (17°C) and summer (22°C) grow-out periods and fed five diets containing graded dietary protein levels of 35, 38, 41, 44 and 47 %. Abalone growth increased with increasing water temperature with weight gains of approximately 100, 280 and 380 % of their initial weight at 12, 17 and 22°C, respectively. Furthermore, the present study clearly demonstrated that higher dietary protein inclusion levels (41 %) than those currently used commercially (35 %) would significantly improve the growth performance when water temperatures are ≥17°C without any adverse impacts on nutrient utilisation, nutrient deposition or nutritional quality of the abalone soft tissue. For example, at 22°C abalone fed a diet containing 41 % protein obtained a significantly higher weight gain percentage (421 %) compared with those fed a diet containing 35 % protein (356 %). Lastly, it is suggested that maintaining a dietary protein inclusion level of 35 % or implementing a 'least cost' feeding approach during cooler seasons, or where water temperatures are ∼12°C, may be beneficial, considering only marginal growth improvements were observed during these periods of slow growth.

Seaweed, also known as macroalgae, represents a vast resource that can be categorized into three taxonomic groups: Rhodophyta (red), Chlorophyta (green), and Phaeophyceae (brown). They are a good source of essential nutrients such as proteins, minerals, vitamins, and omega-3 fatty acids. Seaweed also contains a wide range of functional metabolites, including polyphenols, polysaccharides, and pigments. This study comprehensively discusses seaweed and seaweed-derived metabolites and their potential as a functional feed ingredient in aquafeed for aquaculture production. Past research has discussed the nutritional role of seaweed in promoting the growth performance of fish, but their effects on immune response and gut health in fish have received considerably less attention in the published literature. Existing research, however, has demonstrated that dietary seaweed and seaweed-based metabolite supplementation positively impact the antioxidant status, disease resistance, and stress response in fish. Additionally, seaweed supplementation can promote the growth of beneficial bacteria and inhibit the proliferation of harmful bacteria, thereby improving gut health and nutrient absorption in fish. Nevertheless, an important balance remains between dietary seaweed inclusion level and the resultant metabolic alteration in fish. This review highlights the current state of knowledge and the associated importance of continued research endeavors regarding seaweed and seaweed-based functional metabolites as potential modulators of growth, immune and antioxidant response, and gut microbiota composition in fish.

The production of farmed Atlantic salmon (Salmo salar) is currently linked to finite marine resources, particularly fish oil (FO) and fish meal (FM). Understanding this relationship in a quantitative manner is imperative if this critical balance is to be maintained within sustainable limits as the industry grows. Here we project the potential production and associated growth of the Atlantic salmon aquaculture industry on the basis of a variety of FO and FM utilization scenarios in aquafeed. Reducing FO and FM dietary inclusion to 3% each could permit production growth of 2% per year until the turn of the century (2097 and beyond 2100, respectively), independent of a host of alternatives now being utilized-with three portions of salmon per week providing almost all the recommended weekly long-chain omega-3 fatty acids for human intake. The Atlantic salmon industry's positive annual growth can continue in an era of finite marine resource availability-without the need for additional finite marine resource inputs., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Driven by consumer demand and government policies, synthetic additives in aquafeed require substitution with sustainable and natural alternatives. Seaweeds have been shown to be a sustainable marine source of novel bioactive phenolic compounds that can be used in food, animal and aqua feeds, or microencapsulation applications. For example, phlorotannins are a structurally unique polymeric phenolic group exclusively found in brown seaweed that act through multiple antioxidant mechanisms. Seaweed phenolics show high affinities for binding proteins via covalent and non-covalent bonds and can have specific bioactivities due to their structures and associated physicochemical properties. Their ability to act as protein cross-linkers means they can be used to enhance the rheological and mechanical properties of food-grade delivery systems, such as microencapsulation, which is a new area of investigation illustrating the versatility of seaweed phenolics. Here we review how seaweed phenolics can be used in a range of applications, with reference to their bioactivity and structural properties.

The reduction in benthic light from natural sediment resuspension events, dredging activities and clouds was quantified over multiple time periods (days to weeks) from a 3-year in-situ field study in the inshore turbid-zone coral communities of the Great Barrier Reef. The results were then used to examine the tolerance levels of three coral species and a sponge to light reduction and associated changes in spectral light quality (in conjunction with elevated sediment concentrations) in a 28-day laboratory-based study. All species survived the exposures but sub-lethal responses involving changes in pigmentation, lipids and lipid ratios were observed. A pocilloporid coral was the most sensitive taxon, with a 28-d EC10 value for bleaching (dissociation of the symbiosis) of 2.7 mol photons m 2 d -1 . The possibility of such light reduction levels occurring naturally and/or during maintenance dredging activities was then examined using the 3-year in-situ field study as part of a risk assessment., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Choline plays a crucial role in lipid metabolism for fish, and its deficiency in aquafeed has been linked to compromised health and growth performance. A 56-d experiment was conducted to examine the effects of dietary choline on lipid composition, histology and plasma biochemistry of yellowtail kingfish (Seriola lalandi; YTK; 156 g initial body weight). The dietary choline content ranged from 0·59 to 6·22 g/kg diet. 2-Amino-2-methyl-1-propanol (AMP) (3 g/kg) was added to diets, except for a control diet, to limit de novo choline synthesis. The results showed that the liver lipid content of YTK was similar among diets containing AMP and dominated by NEFA. In contrast, fish fed the control diet had significantly elevated liver TAG. Generally, the SFA, MUFA and PUFA content of liver lipid in fish fed diets containing AMP was not influenced by choline content. The SFA and MUFA content of liver lipid in fish fed the control diet was similar to other diets except for a decrease in PUFA. The linear relationship between lipid digestibility and plasma cholesterol was significant, otherwise most parameters were unaffected. When AMP is present, higher dietary choline reduced the severity of some hepatic lesions. The present study demonstrated that choline deficiency affects some plasma and liver histology parameters in juvenile YTK which might be useful fish health indicators. Importantly, the present study elucidated potential reasons for lower growth in choline-deficient YTK and increased the knowledge on choline metabolism in the fish.

Corals are dependent upon lipids as energy reserves to mount a metabolic response to biotic and abiotic challenges. This study profiled lipids, fatty acids, and microbial communities of healthy and white syndrome (WS) diseased colonies of Acropora hyacinthus sampled from reefs in Western Australia, the Great Barrier Reef, and Palmyra Atoll. Total lipid levels varied significantly among locations, though a consistent stepwise decrease from healthy tissues from healthy colonies (HH) to healthy tissue on WS-diseased colonies (HD; i.e. preceding the lesion boundary) to diseased tissue on diseased colonies (DD; i.e. lesion front) was observed, demonstrating a reduction in energy reserves. Lipids in HH tissues were comprised of high energy lipid classes, while HD and DD tissues contained greater proportions of structural lipids. Bacterial profiling through 16S rRNA gene sequencing and histology showed no bacterial taxa linked to WS causation. However, the relative abundance of Rhodobacteraceae-affiliated sequences increased in DD tissues, suggesting opportunistic proliferation of these taxa. While the cause of WS remains inconclusive, this study demonstrates that the lipid profiles of HD tissues was more similar to DD tissues than to HH tissues, reflecting a colony-wide systemic effect and provides insight into the metabolic immune response of WS-infected Indo-Pacific corals.

Fish are the main source of long-chain polyunsaturated fatty acids (LC-PUFA, >C 18 ) for human consumption. In general, it has been widely observed that the fatty acid (FA) profiles of farmed fish are reflective of the diet. However, the degree of tissue FA "distortion" based on incorporation of different dietary FA into fish tissues varies greatly depending on FA type, fish species and environmental factors. In terms of fish FA composition, this variation has not been comprehensively reviewed, raising the question: "Are fish what they eat?". To date, this remains unanswered in detail. To this end, the present review quantitatively summarized the 'diet-fish' FA relationship via an analysis of FA composition in diets and fish tissues from 290 articles published between 1998 and 2018. Comparison of this relationship among different fish species, tissue types or individual FA was summarized. Furthermore, the influence of environmental factors such as temperature and salinity, as well as of experimental conditions such as fish size and trophic level, feeding duration, and dietary lipid level on this relationship are discussed herein. Moreover, as a means of restoring LC-PUFA in fish, an emphasis was paid to the fish oil finishing strategy after long-term feeding with alternative lipid sources. It is envisaged that the present review will be beneficial in providing a more comprehensive understanding of the fundamental relationship between the FA composition in diets, and subsequently, in the farmed fish. Such information is integral to maintaining the quality of farmed fish fillets from the perspective of FA composition., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study quantified the effects of temperature on reproduction and maternal provisioning of the ectoparasite, Neobenedenia girellae (Platyhelminthes: Monogenea), a species known to cause detrimental impacts to aquaculture fishes in tropical and subtropical environments worldwide. At 20 and 25 °C, parasites exhibited relatively slower production of larger eggs that were energy-dense. In contrast, parasites at 30 °C attained sexual maturity faster, were reproductively active over a shorter period, grew to a smaller size and laid smaller, less energy-rich eggs at a faster rate. As such, parasites exhibited two distinct reproductive patterns in response to temperature: parasites at lower temperatures produced larger eggs with higher energy content, while those at the higher temperature had a higher rate of egg production. Larger eggs produced under cooler conditions were better provisioned with energetic reserves and important, membrane-bound lipids that would likely facilitate larval longevity and development success. This is commensurate with previous observations of epizootics of this parasite species in aquaculture systems during winter. Meanwhile, eggs produced at 30 °C contained higher proportions of saturated fatty acids compared with polyunsaturated fatty acids, likely reflecting metabolic regulation of cell membrane fluidity, which is necessary for larvae to survive warm conditions. This study demonstrates that fish ectoparasites have evolved substantial reproductive and metabolic flexibility to maximise infection success under variable environmental conditions., (Copyright © 2020. Published by Elsevier Ltd.)

Tamoxifen (TAM) is an antiestrogenic agent and can enter the aquatic environment in wastewater. It has been reported that TAM can induce hepatic steatosis in vertebrates, however, the effects of TAM exposure on lipid metabolism of hepatopancreas in crustaceans remains unclear. In this study, four TAM concentrations (0, 6.7, 13.4 and 20 μg g -1 crab body weight) were injected into the swimming-leg of swimming crabs Portunus trituberculatus, as a means of evaluating the effects of TAM on the expression levels of lipid metabolism-related genes, lipid composition, and hepatopancreas histology. The results showed that the mRNA levels of three lipogenic related genes (diacylglycerol acyltransferase 1 (DGAT1), acetyl-CoA carboxylase (ACC) and fatty acyl desaturase (FAD)) decreased significantly in the 6.7 μg g -1 and 20 μg g -1 TAM treatments compare to the control. The mRNA levels of fatty acid synthase (FAS) decreased significantly in a dose-dependent manner as TAM concentration increased. The mRNA levels of two lipid catabolism-related genes (acyl-CoA oxidase (ACOX) and fatty acid transport protein (FATP)) were down-regulated among the three TAM treatments, while the enzyme activity and mRNA level of carnitine palmitoyltransferase I (CPT-I) was up-regulated by TAM treatments. Compared to the control, the lowest levels of total lipids and phospholipids were detected in the 6.7 μg g -1 TAM treatment, while the 20 μg g -1 TAM treatment had the lowest free fatty acids concentration. The 6.7 μg g -1 TAM treatment had the lowest percentages of 16:1n-7, 18:1n-9, 18:1n-7 and total monounsaturated fatty acids (∑MUFA), whilst simultaneously recording the highest percentages of 18:2n-6 and 20:2n-6 in this treatment. Moreover, histological observations indicated that TAM caused the walls of the hepatopancreatic tubules to become brittle, with a concurrent increase in the number of blister-like cells. These results suggest that TAM damages the hepatopancreas and leads to a reduction in hepatopancreatic lipid deposition in P. trituberculatus., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 β-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.

Developing an optimal heterotrophic feeding regime has the potential to improve captive coral growth and health. This study evaluated the efficacy of three exogenous diets: Artemia nauplii (ART), a commercially available coral diet (Reef Roids) (RR), and a novel, micro-bound diet (ATF), against a comparatively natural, unfiltered seawater treatment (RAW), and an unfed, ultra-filtered seawater treatment (CTL), in adult Acropora millepora and Pocillopora acuta nubbins. After 90 days, both species showed significantly positive weight gain in response to one treatment (A. millepora-RAW, P. acuta-ART), and comparatively low growth in response to another (A. millepora-ATF, P. acuta-RR). The results highlighted substantial differences in the nutritional requirements between species. The nutritional composition of A. millepora in the best performing treatment was dominated by high-energy materials such as storage lipids and saturated and monounsaturated fatty acids. In contrast, the P. acuta nutritional profile in the superior treatment showed a predominance of structural materials, including protein, phospholipids, and polyunsaturated fatty acids. This study demonstrates that Artemia nauplii can successfully replace a natural feeding regime for captive P. acuta, yet highlights the considerable work still required to optimise supplementary feeding regimes for A. millepora., Competing Interests: The authors have declared that no competing interests exist.

Ocean warming (OW) and ocean acidification (OA) are threatening coral reef ecosystems, with a bleak future forecast for reef-building corals, which are already experiencing global declines in abundance. In contrast, many coral reef sponge species are able to tolerate climate change conditions projected for 2100. To increase our understanding of the mechanisms underpinning this tolerance, we explored the lipid and fatty acid (FA) composition of four sponge species with differing sensitivities to climate change, experimentally exposed to OW and OA levels predicted for 2100, under two CO 2 Representative Concentration Pathways. Sponges with greater concentrations of storage lipid, phospholipids, sterols and elevated concentrations of n-3 and n-6 long-chain polyunsaturated FA (LC PUFA), were more resistant to OW. Such biochemical constituents likely contribute to the ability of these sponges to maintain membrane function and cell homeostasis in the face of environmental change. Our results suggest that n-3 and n-6 LC PUFA are important components of the sponge stress response potentially via chain elongation and the eicosanoid stress-signalling pathways. The capacity for sponges to compositionally alter their membrane lipids in response to stress was also explored using a number of specific homeoviscous adaptation (HVA) indicators. This revealed a potential mechanism via which additional CO 2 could facilitate the resistance of phototrophic sponges to thermal stress through an increased synthesis of membrane-stabilizing sterols. Finally, OW induced an increase in FA unsaturation in phototrophic sponges but a decrease in heterotrophic species, providing support for a difference in the thermal response pathway between the sponge host and the associated photosymbionts. Here we have shown that sponge lipids and FA are likely to be an important component of the sponge stress response and may play a role in facilitating sponge survival under future climate conditions., (© 2018 John Wiley & Sons Ltd.)

Marine ecosystems are responsible for virtually all production of omega-3 (ω3) long-chain polyunsaturated fatty acids (PUFA), which are essential nutrients for vertebrates. Current consensus is that marine microbes account for this production, given their possession of key enzymes including methyl-end (or "ωx") desaturases. ωx desaturases have also been described in a small number of invertebrate animals, but their precise distribution has not been systematically explored. This study identifies 121 ωx desaturase sequences from 80 species within the Cnidaria, Rotifera, Mollusca, Annelida, and Arthropoda. Horizontal gene transfer has contributed to this hitherto unknown widespread distribution. Functional characterization of animal ωx desaturases provides evidence that multiple invertebrates have the ability to produce ω3 PUFA de novo and further biosynthesize ω3 long-chain PUFA. This finding represents a fundamental revision in our understanding of ω3 long-chain PUFA production in global food webs, by revealing that numerous widespread and abundant invertebrates have the endogenous capacity to make significant contributions beyond that coming from marine microbes.

Scleractinian corals are colonial organisms comprising multiple physiologically integrated polyps and branches. Colonialism in corals is highly beneficial, and allows a single colony to undergo several life processes at once through physiological integration and compartmentalised functioning. Elucidating differences in the biochemical composition of intra-colonial branch positions will provide valuable insight into the nutritional reserves underlying different regions in individual coral colonies. This will also ascertain prudent harvesting strategies of wild donor-colonies to generate coral stock with high survival and vigour prospects for reef-rehabilitation efforts and captive husbandry. This study examined the effects of colony branch position on the nutritional profile of two different colony sizes of the common scleractinian, Acropora millepora . For smaller colonies, branches were sampled at three locations: the colony centre (S-centre), 50% of the longitudinal radius length (LRL) (S-50), and the colony edge (S-edge). For larger colonies, four locations were sampled: the colony centre (L-centre), 33.3% of the LRL (L-33), 66.6% of the LRL (L-66), and the edge (L-edge). Results demonstrate significant branch position effects, with the edge regions containing higher protein, likely due to increased tissue synthesis and calcification. Meanwhile, storage lipid and total fatty acid concentrations were lower at the edges, possibly reflecting catabolism of high-energy nutrients to support proliferating cells. Results also showed a significant effect of colony size in the two classes examined. While the major protein and structural lipid sink was exhibited at the edge for both sizes, the major sink for high-energy lipids and fatty acids appeared to be the L-66 position of the larger colonies and the S-centre and S-50 positions for the smaller colonies. These results confirm that the scleractinian coral colony is not nutritionally homogeneous, and while different regions of the coral colony are functionally specialised, so too are their nutritional profiles geared toward meeting specific energetic demands., Competing Interests: The authors declare there are no competing interests.

This study on the eggs of the tropical monogenean Neobenedenia girellae presents the first detailed quantitative biochemical information of a marine parasite species' eggs. Moisture and protein composed the majority of the contents of freshly laid eggs (79.12±0.82 and 11.51±0.49% respectively) followed by lipid (2.50±0.15%). Lipids were composed of approximately equal amounts of saturated, monounsaturated, and polyunsaturated fatty acids and the predominant lipid class was triacylglycerol (33.82±1.20%). This study represents a fundamental step towards a better understanding of the early life biology of this important species of parasite., (Copyright © 2017. Published by Elsevier B.V.)

Heterotrophic feeding in newly-settled coral planulae can potentially improve survivorship and accelerate early development in some species; however, an optimal diet to facilitate this does not currently exist. This study evaluated the efficacy of three heterotrophic feeding regimes (enriched rotifers, unfiltered seawater, and a novel, particulate diet), against a wholly-phototrophic treatment on Acropora hyacinthus, A. loripes, A. millepora, and A. tenuis recruits, over 93 days post-settlement. The unfiltered seawater treatment recorded maximum survival for all species (A. hyacinthus 95.9±8.0%, A. loripes: 74.3±11.5%, A. millepora: 67±12.7%, A. tenuis: 53.2±11.3%), although not significant. Growth (% surface area gain) was also greatest in the unfiltered seawater, and this was significant for A. millepora (870±307%) and A. tenuis (693±91.8%) (p<0.05). Although total lipid concentration was relatively stable across treatments, the lipid class composition exhibited species-specific responses to each treatment. Lower saturated and higher polyunsaturated fatty acids appeared beneficial to recruit performance, particularly in the unfiltered seawater, which generally contained the highest levels of 20:5n-3 (EPA), 22:6n-3 (DHA), and 20:4n-6 (ARA). The present study demonstrates the capacity of a nutritionally adequate and readily accepted heterotrophic feeding regime to increase coral recruit survival, growth, and health, which can greatly reduce the time required in cost- and labour-intensive culture.

Lipids are involved in a host of biochemical and physiological processes in corals. Therefore, changes in lipid composition reflect changes in the ecology, nutrition, and health of corals. As such, accurate lipid extraction, quantification, and identification is critical to obtain comprehensive insight into a coral's condition. However, discrepancies exist in sample preparation methodology globally, and it is currently unknown whether these techniques generate analogous results. This study compared the two most common sample preparation techniques for lipid analysis in corals: (1) tissue isolation by air-spraying and (2) crushing the coral in toto . Samples derived from each preparation technique were subsequently analysed to quantify lipids and their constituent classes and fatty acids in four common, scleractinian coral species representing three distinct morphotypes ( Acropora millepora , Montipora crassotuberculata , Porites cylindrica , and Pocillopora damicornis ). Results revealed substantial amounts of organic material, including lipids, retained in the skeletons of all species following air-spraying, causing a marked underestimation of total lipid concentration using this method. Moreover, lipid class and fatty acid compositions between the denuded skeleton and sprayed tissue were substantially different. In particular, the majority of the total triacylglycerol and total fatty acid concentrations were retained in the skeleton (55-69% and 56-64%, respectively). As such, the isolated, sprayed tissue cannot serve as a reliable proxy for lipid quantification or identification in the coral holobiont. The in toto crushing method is therefore recommended for coral sample preparation prior to lipid analysis to capture the lipid profile of the entire holobiont, permitting accurate diagnoses of coral condition., Competing Interests: The authors declare there are no competing interests.

This study explores how plasticity in biochemical attributes, used as indicators of health and condition, enables the coral Acropora tenuis to respond to differing water quality regimes in inshore regions of the Great Barrier Reef. Health attributes were monitored along a strong and weak water quality gradient, each with three reefs at increasing distances from a major river source. Attributes differed significantly only along the strong gradient; corals grew fastest, had the least dense skeletons, highest symbiont densities and highest lipid concentrations closest to the river mouth, where water quality was poorest. High nutrient and particulate loads were only detrimental to skeletal density, which decreased as linear extension increased, highlighting a trade-off. Our study underscores the importance of assessing multiple health attributes in coral reef monitoring. For example, autotrophic indices are poor indicators of coral health and condition, but improve when combined with attributes like lipid content and biomass., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Nutritional strategies are currently developed to produce farmed fish rich in n-3 long-chain PUFA (LC-PUFA) whilst replacing fish oil by plant-derived oils in aquafeeds. The optimisation of such strategies requires a thorough understanding of fish lipid metabolism and its nutritional modulation. The present study evaluated the fatty acid bioconversion capacity of rainbow trout (Oncorhynchus mykiss) fry previously depleted in n-3 PUFA through a 60-d pre-experimental feeding period with a sunflower oil-based diet (SO) followed by a 36-d experimental period during which fish were fed either a linseed oil-based diet (LO) (this treatment being called SO/LO) or a fish oil-based diet (FO) (this treatment being called SO/FO). These treatments were compared with fish continuously fed on SO, LO or FO for 96 d. At the end of the 36-d experimental period, SO/LO and SO/FO fish recovered >80 % of the n-3 LC-PUFA reported for LO and FO fish, respectively. Fish fed on LO showed high apparent in vivo elongation and desaturation activities along the n-3 biosynthesis pathway. However, at the end of the experimental period, no impact of the fish n-3 PUFA depletion was observed on apparent in vivo elongation and desaturation activities of SO/LO fish as compared with LO fish. In contrast, the fish n-3 PUFA depletion negatively modulated the n-6 PUFA bioconversion capacity of fish in terms of reduced apparent in vivo elongation and desaturation activities. The effects were similar after 10 or 36 d of the experimental period, indicating the absence of short-term effects.

In this study, we report the distribution of orexin A (OXA), orexin B (OXB), and orexin receptor (OX2R) immunoreactive (ir) cells in the hypothalamus and gastrointestinal tract of Oncorhynchus mykiss fed diets with different dietary fatty acid compositions. Trout were fed five iso-energetic experimental diets containing fish oil, or one of four different vegetable oils (olive, sunflower, linseed, and palm oils) as the added dietary lipid source for 12 weeks. OXA, OXB, and OX2R immunoreactive neurons and nervous fibers were identified in the lateral and ventro-medial hypothalamus. OXA, OXB, and OX2R ir cells were found in the mucosa and glands of the stomach and in the mucosa of both the pyloric cecae and intestine. OX2R ir cells were localized in the mucosa layer of both the pyloric cecae and intestine. These immunohistochemical (IHC) results were confirmed via Western blotting. Antibodies against preproorexin (PPO) crossreacted with a band of ∼16 kDa in the hypothalamus, stomach, pyloric cecae, and intestine. Antibodies against OX2R crossreacted with a band of ∼38 kDa in the hypothalamus, pyloric cecae, and intestine. The presence and distribution of OXA, OXB, and OX2R ir cells in the hypothalamus and gastrointestinal tract did not appear to be affected by dietary oils. The presence of orexin system immunoreactive cells in the stomach, pyloric cecae, and intestine of rainbow trout, but not in the enteric nervous system, could suggest a possible role of these peptides as signaling of gastric emptying or endocrine modulation, implying a main local action played by orexins., (© 2015 Wiley Periodicals, Inc.)

It is known that fatty acids (FA) regulate lipid metabolism by modulating the expression of numerous genes. In order to gain a better understanding of the effect of individual FA on lipid metabolism related genes in rainbow trout (Oncorhynchus mykiss), an in vitro time-course study was implemented where twelve individual FA (butyric 4:0; caprylic 8:0; palmitic (PAM) 16:0; stearic (STA) 18:0; palmitoleic16:1n-7; oleic 18:1n-9; 11-cis-eicosenoic 20:1n-9; linoleic (LNA) 18:2n-6; α-linolenic (ALA) 18:3n-3; eicosapentenoic (EPA) 20:5n-3; docosahexaenoic (DHA) 22:6n-3; arachidonic (ARA) 20:4n-6) were incubated in rainbow trout liver slices. The effect of FA administration over time was evaluated on the expression of leptin, PPARα and CPT-1 (lipid oxidative related genes). Leptin mRNA expression was down regulated by saturated fatty acids (SFA) and LNA, and was up regulated by monounsaturated fatty acids (MUFA) and long chain PUFA, whilst STA and ALA had no effect. PPARα and CPT-1mRNA expression were up regulated by SFA, MUFA, ALA, ARA and DHA; and down regulated by LNA and EPA. These results suggest that there are individual and specific FA induced modifications of leptin, PPARα and CPT-1 gene expression in rainbow trout, and it is envisaged that such results may provide highly valuable information for future practical applications in fish nutrition.

Largely attributable to concerns surrounding sustainability, the utilisation of omega-3 long-chain polyunsaturated fatty acid-rich (n-3 LC-PUFA) fish oils in aquafeeds for farmed fish species is an increasingly concerning issue. Therefore, strategies to maximise the deposition efficiency of these key health beneficial fatty acids are being investigated. The present study examined the effects of four vegetable-based dietary lipid sources (linseed, olive, palm and sunflower oil) on the deposition efficiency of n-3 LC-PUFA and the circulating blood plasma concentrations of the appetite-regulating hormones, leptin and ghrelin, during the grow-out and finishing phases in rainbow trout culture. Minimal detrimental effects were noted in fish performance; however, major modifications were apparent in tissue fatty acid compositions, which generally reflected that of the diet. These modifications diminished somewhat following the fish oil finishing phase, but longer-lasting effects remained evident. The fatty acid composition of the alternative oils was demonstrated to have a modulatory effect on the deposition efficiency of n-3 LC-PUFA and on the key endocrine hormones involved in appetite regulation, growth and feed intake during both the grow-out and finishing phases. In particular, n-6 PUFA (sunflower oil diet) appeared to 'spare' the catabolism of n-3 LC-PUFA and, as such, resulted in the highest retention of these fatty acids, ultimately highlighting new nutritional approaches to maximise the maintenance of the qualitative benefits of fish oils when they are used in feeds for aquaculture species.

Monola oil, a high oleic acid canola cultivar, and canola oil were evaluated as replacers of fish oil at three levels of inclusion (60%, 75% and 90%) in rainbow trout diets. After a 27-week grow-out cycle, the diet-induced effects on growth, fatty acid metabolism and final eating quality were assessed. Overall, no effects were noted for growth, feed utilisation or fish biometry, and the fatty acid composition of fish fillets mirrored that of the diets. Dietary treatments affected fillet lipid oxidation (free malondialdehyde), pigmentation and flavour volatile compounds, but only minor effects on sensorial attributes were detected. Ultimately, both oils were demonstrated to possess, to differing extents, suitable qualities to adequately replace fish oil from the perspective of fish performance and final product quality. However, further research is required to alleviate on-going issues associated with the loss of health promoting attributes (n-3 long chain polyunsaturated fatty acids) of final farmed products., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The implementation of alternative lipid sources for use in aquaculture is of considerable interest globally. However, the possible benefit of using stearidonic acid (SDA)-rich fish oil (FO) alternatives has led to scientific confusion. Two hundred and forty rainbow trout (Oncorhynchus mykiss) were fed 1 of 4 diets (3 replicate tanks/treatment) containing either FO, linseed oil (LO), echium oil, or mixed vegetable oil (72% LO, 23% sunflower oil, and 6% canola oil) as the dietary lipid source (16.5%) for 73 d to investigate the competition and long-chain PUFA (LC-PUFA) biosynthesis between the fatty acid substrates α-linolenic acid (ALA) and SDA. SDA was more efficiently bioconverted to LC-PUFA compared with ALA. However, when the dietary lipid sources were directly compared, the increased provision of C18 PUFA within the LO diet resulted in no significant differences in (n-3) LC-PUFA content compared with fish fed the other diets. This study therefore shows that, rather than the previously speculated substrate competition, the limiting process in the apparent in vivo (n-3) LC-PUFA biosynthesis appears to be substrate availability. Rainbow trout fed the SDA- and ALA-rich dietary lipid sources subsequently had similar significant reductions in (n-3) LC-PUFA compared with fish fed the FO diet, therefore providing no additional dietary benefit on (n-3) LC-PUFA concentrations.

Fish oil replacement in aquaculture feeds results in major modifications to the fatty acid makeup of cultured fish. Therefore, in vivo fatty acid biosynthesis has been a topic of considerable research interest. Evidence suggests that pyridoxine (vitamin B(6)) plays a role in fatty acid metabolism, and in particular, the biosynthesis of LC-PUFA has been demonstrated in mammals. However, there is little information on the effects of dietary pyridoxine availability in fish fed diets lacking LC-PUFA. This study demonstrates a relationship between dietary pyridoxine supplementation and fatty acid metabolism in rainbow trout. In particular, the dietary pyridoxine level was shown to modulate and positively stimulate the activity of the fatty acid elongase and Δ-6 and Δ-5 desaturase enzymes, deduced by the whole-body fatty acid balance method. This activity was insufficient to compensate for a diet lacking in LC-PUFA but does highlight potential strategies to maximize this activity in cultured fish, especially when fish oil is replaced with vegetable oils.

Five experimental diets with constant total C(18) PUFA and varying 18:3n-3/18:2n-6 ratios were fed to rainbow trout over an entire production cycle. The whole-body fatty acid balance method demonstrated a clear trend of progressively reduced fatty acid bioconversion activity along the n-3 and n-6 pathways, up to the production of 20:5n-3 and 20:4n-6, respectively. This suggests that the pathway exhibits a "funnel like" progression of activity rather than the existence of a single rate limiting step. The production of 22:5n-3 and 22:6n-3 was more active than that of 20:5n-3. However, despite this trend in reduced apparent in vivo net enzyme activity, the efficiency of the various bioconversion steps (measured as % of bioconverted substrate) confirmed an opposing trend. A 3.2-fold higher Δ-6 desaturase affinity towards 18:3n-3 over 18:2n-6 and an 8-fold greater Δ-5 desaturase affinity towards 20:4n-3 over 20:3n-6 were recorded. The main results of the study were that (1) rainbow trout are quite efficient at bioconverting 18:3n-3 to 22:6n-3, and (2) the LC-PUFA biosynthetic pathway is substrate limited. Fillet n-3 LC-PUFA concentrations increased with the increasing dietary supply of 18:3n-3. Despite an almost identical dietary supply of n-3 LC-PUFA, originating from the fish meal fraction of the diets, the fillets of trout fed the diet richest in 18:3n-3 were 2-fold higher in n-3 LC-PUFA than fish fed low 18:3n-3 diets. Nevertheless, fillets of trout fed a fish oil control diet contained more than double the amount of n-3 LC-PUFA compared to fish fed the diets richest in 18:3n-3.

Global shortages in fish oil are forcing the aquaculture feed industry to use alternative oil sources, the use of which negatively affects the final fatty acid makeup of cultured fish. Thus, the modulation of fatty acid metabolism in cultured fish is the core of an intensive global research effort. The present study aimed to evaluate the effects of various dietary α-linolenic acid (ALA, 18:3n-3)/linoleic acid (LA, 18:2n-6) ratios in cultured fish. A feeding trial was implemented on the freshwater finfish Murray cod, in which fish were fed either a fish oil-based control diet or one of five fish oil-deprived experimental diets formulated to contain an ALA/LA ratio ranging from 0.3 to 2.9, but with a constant total C₁₈ PUFA (ALA+LA) content. The whole-body fatty acid balance method was used to evaluate fish in vivo fatty acid metabolism. The results indicate that dietary ALA was more actively β-oxidized and bioconverted, whereas LA appears to be more efficiently deposited. LA was β-oxidized at a constant level (~36% of net intake) independent of dietary availability, whereas ALA was oxidized proportionally to dietary supply. The in vivo apparent Δ-6 desaturase activity on n-3 and n-6 PUFA exhibited an increasing and decreasing trend, respectively, in conjunction with the increasing dietary ALA/LA ratio, clearly indicating that this enzymatic activity is substrate dependent. However, the maximum Δ-6 desaturase activity acting on ALA peaked at the substrate level of 3.2186 (μmol g fish⁻¹ day⁻¹), suggesting that additional inclusion of ALA is not only wasteful but counterproductive in terms of n-3 LC-PUFA production. Despite a constant total supply of ALA+LA, the recorded total in vivo apparent Δ-6 desaturase activity on both substrates (ALA and LA) increased in synchrony with the ALA/LA ratio, peaking at 1.54, and a 3.2-fold greater Δ-6 desaturase affinity toward ALA over LA was recorded.

In consideration of economical and environmental concerns, fish oil (FO) substitution in aquaculture is the focus of many fish nutritionists. The most stringent drawback of FO replacement in aquafeeds is the consequential modification to the final fatty acid (FA) make-up of the fish fillet. However, it is envisaged that a solution may be achieved through a better understanding of fish FA metabolism. Therefore, the present study investigated the fate of individual dietary FA in rainbow trout (Oncorhynchus mykiss) fed a FO-based diet (rich in 20 : 5n-3) or a linseed oil-based diet (LO; rich in 18 : 3n-3). The study demonstrated that much of the 18 : 3n-3 content from the LO diet was oxidised and, despite the significantly increased accretion of Delta-6 and Delta-5 desaturated FA, a 2- and 3-fold reduction in the fish body content of 20 : 5n-3 and 22 : 6n-3, respectively, compared with the FO-fed fish, was recorded. The accretion of longer-chain FA was unaffected by the dietary treatments, while there was a greater net disappearance of FA provided in dietary surplus. SFA and MUFA recorded a net accretion of FA produced ex novo. In the fish fed the FO diet, the majority of dietary 20 : 5n-3 was accumulated (53.8 %), some was oxidised (14.7 %) and a large proportion (31.6 %) was elongated and desaturated up to 22 : 6n-3. In the fish fed the LO diet, the majority of dietary 18 : 3n-3 was accumulated (58.1 %), a large proportion was oxidised (29.5 %) and a limited amount (12.4 %) was bio-converted to longer and more unsaturated homologues.

Dietary fatty acids are known to modulate fatty acid metabolism in fish. However, the innate capability of fish to bioconvert short chain fatty acids to health promoting long chain fatty acids (LCPUFA) is insufficient to compensate for a reduced dietary intake. While many studies have focused on the dietary regulation of the fatty acid bioconversion pathways, there is little known regarding the effects of the dietary levels of C(18) polyunsaturated fatty acids (PUFA) on fatty acid metabolism. Here, we show a greater degree of apparent enzyme activity (Δ-6 desaturase) in fish fed a diet with higher amounts of dietary C(18) PUFA. In particular, fish receiving high amounts of dietary C(18) PUFA had a greater amount of Δ-6 desaturase activity acting on 18:3n-3 than 18:2n-6. However, with the gradual reduction of dietary C(18) PUFA there was a shift in substrate preference of Δ-6 desaturase from 18:3n-3 to 18:2n-6. This information will provide valuable insight for the implementation of low fish oil diets, which permit the maintenance of n-3 LCPUFA levels in farmed Murray cod.

Currently there are several contrasting methods utilized for estimating elongation and desaturation of fatty acids and their general metabolism. The majority of these methods involve an ex vivo approach, requiring expensive and sophisticated equipment, likely to result in considerable variation in enzyme activity between and within species. In the present paper we introduce a further development of the whole-body fatty acid balance method for the estimation of the elongation and desaturation of fatty acids. This method though receiving considerable attention because of its simplicity and reliability has yet to be presented in detail. Theoretically, the whole-body fatty acid balance method can potentially be applied to any organism and requires little more than a gas chromatography unit for fatty acid analysis and elementary calculations. As such in this paper we attempt to spell out in detail the theoretical basis and the methods of application drawing specific examples. Using the present method it is possible to measure the fate of individual fatty acids towards desaturation, elongation and oxidation and calculate the elongase, Delta-6 desaturase and Delta-5 desaturase activities.

The aim of the present investigation was to quantify the fate of C18 and long chain polyunsaturated dietary fatty acids in the freshwater fish, Murray cod, using the in vivo, whole-body fatty acid balance method. Juvenile Murray cod were fed one of five iso-nitrogenous, iso-energetic, semipurified experimental diets in which the dietary fish oil (FO) was replaced (0, 25, 50, 75, and 100%) with a blended vegetable oil (VO), specifically formulated to match the major fatty acid classes [saturated fatty acids, monounsaturated fatty acids, n-3 polyunsaturated fatty acids (PUFA), and n-6 PUFA] of cod liver oil (FO). However, the PUFA fraction of the VO was dominated by C18 fatty acids, while C20/22 fatty acids were prevalent in the FO PUFA fraction. Generally, there was a clear reflection of the dietary fatty acid composition across each of the five treatments in the carcass, fillet, and liver. Lipid metabolism was affected by the modification of the dietary lipid source. The desaturation and elongation of C18 PUFAs increased with vegetable oil substitution, supported by the occurrence of longer and higher desaturated homologous fatty acids. However, increased elongase and desaturase activity is unlikely to fulfill the gap observed in fatty acid composition resulting from decreased highly unsaturated fatty acids intake.

The whole-body fatty acid balance method was used to investigate the fatty acid metabolism in Murray cod (Maccullochella peelii peelii) fed diets containing canola (CO) or linseed oil (LO). Murray cod were able to elongate and desaturate both 18:2n-6 and 18:3n-3. In fish fed the CO diet, 54.4% of the 18:2n-6 consumed was accumulated, 38.5% oxidized and 6.4% elongated and desaturated to higher homologs. Fish fed the LO diet accumulated 52.9%, oxidized 37% and elongated and desaturated 8.6% of the consumed 18:3n-3. The overall roles of n-6 fatty acids appeared more important in Murray cod compared to other freshwater species. Murray cod also showed a preferential order of utilization of C18 fatty acid for energy production (18:3n-3 > 18:2n-6 > 18:1n-9). Moreover, it is demonstrated that an increase in dietary 18:3n-3 is directly responsible of increased desaturase activity and augmented saturated fatty acid accumulation in the fish body. The present study also suggests that, in the context of the possible maximization of the natural ability of fish to produce long chain polyunsaturated fatty acids, the whole-body approach can be considered well suited and informative and Murray cod is a suited candidate to fish oil replacement for its diets.