Arachidonic acid and eicosapentaenoic acid metabolism in juvenile Atlantic Salmon as affected by water temperature

Salmons raised in aquaculture farms around the world are increasingly subjected to sub-optimal environmental conditions, such as high water temperatures during summer seasons. Aerobic scope increases and lipid metabolism changes are known plasticity responses of fish for a better acclimation to high water temperature. The present study aimed at investigating the effect of high water temperature on the regulation of fatty acid metabolism in juvenile Atlantic salmon fed different dietary ARA/EPA ratios (arachidonic acid, 20:4n-6/ eicosapentaenoic acid, 20:5n-3), with particular focus on apparent in vivo enzyme activities and gene expression of lipid metabolism pathways. Three experimental diets were formulated to be identical, except for the ratio EPA/ARA, and fed to triplicate groups of Atlantic salmon (Salmo salar) kept either at 10°C or 20°C. Results showed that fatty acid metabolic utilisation, and likely also their dietary requirements for optimal performance, can be affected by changes in their relative levels and by environmental temperature in Atlantic salmon. Thus, the increase in temperature, independently from dietary treatment, had a significant effect on the β-oxidation of a fatty acid including EPA, as observed by the apparent in vivo enzyme activity and mRNA expression of pparα -transcription factor in lipid metabolism, including β-oxidation genes- and cpt1 -key enzyme responsible for the movement of LC-PUFA from the cytosol into the mitochondria for β-oxidation-, were both increased at the higher water temperature. An interesting interaction was observed in the transcription and in vivo enzyme activity of Δ5fad-time-limiting enzyme in the biosynthesis pathway of EPA and ARA. Such, at lower temperature, the highest mRNA expression and enzyme activity was recorded in fish with limited supply of dietary EPA, whereas at higher temperature these were recorded in fish with limited ARA supply. In consideration that fish at higher water temperature recorded a significantly increased feed intake, these results clearly suggested that at high, sub-optimal water temperature, fish metabolism attempted to increment its overall ARA status -the most bioactive LC-PUFA participating in the inflammatory response- by modulating the metabolic fate of dietary ARA (expressed as % of net intake), reducing its β-oxidation and favouring synthesis and deposition. This correlates also with results from other recent studies showing that both immune- and stress- responses in fish are up regulated in fish held at high temperatures. This is a novel and fundamental information that warrants industry and scientific attention, in consideration of the imminent increase in water temperatures, continuous expansion of aquaculture operations, resources utilisation in aquafeed and much needed seasonal/adaptive nutritional strategies.

Can dietary lipid source circadian alternation improve omega-3 deposition in rainbow trout?

With the salmonid industry currently exploiting the vast majority of globally available fish oil, there is the need to optimise fish oil utilisation by increasing its efficiency in terms of transferring the health-promoting long chain omega-3 fatty acids (n−3 LC-PUFA) into farmed fish flesh. The aim of this study was to evaluate if dietary fatty acid deposition is affected by the time of feeding, and hence identify possible innovative feeding strategies towardsmore efficient use of dietary fish oil. Over a period of 12 weeks, three diets with different lipid sources, canola oil (CO), fish oil (FO) or a 50/50 blend of the two oils (Mix), were alternated daily and fed to rainbow trout (Oncorhynchus mykiss). Six treatments were administered to fish, reference treatment (REF, continuously fed FO), control treatment (CT, continuously fed Mix), am canola oil ration (amCOR), pm canola oil ration (pmCOR), am canola oil satiation (amCOS) and pm canola oil satiation (pmCOS). Fish received either the CO diet in the am or pm feeds and received the FO diet at the opposite time. A significant increase in growth and feed consumption was noted in the pmCOS treatment. Fillet fatty acid profile was modified by associated feeding schedules and was generally reflective of dietary fatty acid profile. No significant increases in n−3 LCPUFA deposition were observed. However, both linoleic acid (18:2n−6) and α-linolenic acid (18:3n−3) contents were significantly higher in pmCOR compared to amCOR and CT. The results of the present study suggest the existence of cyclical circadian patterns in fatty acid deposition in rainbow trout.

n-3 LC-PUFA deposition efficiency and appetite regulating hormones are modulated by the dietary lipid source during rainbow trout grow-out and finishing periods

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.

From individuals to populations : prey fish risk-taking mediates mortality in whole-system experiments

Recent research suggests that the behavior of individuals under risk of predation could be a key link between individual behavior and population and community dynamics. Yet existing theory remains largely untested at large spatial and temporal scales. We manipulated food available to age-0 rainbow trout while at risk of cannibalism, in a replicated factorial whole-lake experiment, to test whether the trade-off between growth and mortality rates is mediated by foraging activity by young fish under predation risk. We found that this trade-off exists for young fish at the whole-system scale, and that food-dependent behavioral variation has large mortality consequences. In high-food lakes, age-0 trout spent less time moving, fewer individuals swam continuously, and those swimming continuously swam at slower speeds relative to those in low-food lakes. Age-0 trout also used deep, risky habitats less when food was abundant. This lower activity, combined with avoidance of risky habitats, coincided with 68% higher survival in high-food lakes. If general, this trade-off may be a key mechanism linking individual behavior to population-level processes in size-structured populations.

Dietary n-6/n-3 LC-PUFA ratio, temperature and time interactions on nutrients and fatty acids digestibility in Atlantic salmon

The objective of the present study was to evaluate the effects of altered dietary n-3/n-6 LC-PUFA ratio, adaptation to diet over time, different water temperatures, and their interactions on nutrients and fatty acids digestibility in juvenile Atlantic salmon. Three experimental diets were formulated to be identical, with the only exception of the ratio of eicosapentaenoic acid (EPA, 20:5n-3) to arachidonic acid (ARA, 20:4n-6), and fed to triplicate groups of juvenile Atlantic salmon (Salmo salar) of 55. g initial body weight. Fish were reared in a fully controlled recirculating aquaculture system, fed to apparent satiety twice daily and kept at 10. °C and for an initial period of 100. days, and faeces were collected for digestibility estimation. Then, half of the fish of each experimental tank were moved to a separate system, where the water temperature was gradually increased up to 20. °C. Fish were maintained in the two systems for an additional period of 50. days, and faeces were collected for digestibility estimation from both groups of fish at the two water temperatures. This study concluded that dietary treatments and time had only minor effects, whereas environmental temperature resulted in modified digestibility values, with increased nutrient digestibility with increasing temperature. Varying EPA/ARA ratio in the diet had only minor direct effects on digestibility, with no direct effect on overall nutrients digestibility, and fundamentally only statistically significant effects in the fatty acid digestibility of EPA and ARA themselves. Because of current increasing pressure for more efficient fish oil replacement strategies, increasing interest in dietary ARA in aquafeed and increasing relevance and occurrence of sub-optimal rearing temperature in commercial aquaculture, this study can be considered to be important as it provided a series of fundamental information, which are envisaged to be useful towards addressing these constraints and possible nutritional remedial strategies.

Marginal efficiencies of long chain-polyunsaturated fatty acid use by barramundi (Lates calcarifer) when fed diets with varying blends of fish oil and poultry fat

An experiment was conducted with barramundi (Lates calcarifer) juveniles to examine the marginal efficiency of utilisation of long chain-polyunsaturated fatty acids (LC-PUFA). A series of five diets with blends of fish (anchovy) oil and poultry fat (F100:P0, F60:P40, F30:P70, F15:P85, F0:P100) were fed to 208. ±. 4.1. g fish over a 12-week period. The replacement of fish oil with poultry fat had no impact on growth performance (average final weight of 548.3. ±. 10.2. g) or feed conversion (mean = 1.14. ±. 0.02). Analysis of the whole body composition showed that the fatty acid profile reflected that of the fed diet. However it was also shown that there was a disproportional retention of some fatty acids relative to others (notably LOA, 18:2n-6 and LNA, 18:3n-3). By examining the body mass independent retention of different fatty acids with differential levels of intake of each, the marginal efficiencies of the use of these nutrients by this species were able to be determined. The differential retention of fatty acids in the meat was also examined allowing the determination of oil blending strategies to optimise meat n-3 LC-PUFA levels.

Chemical changes in fed and starved larval trout cod, Maccullochella macquarensis during early development

The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in μmoles larva^–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 μg mg lipid^–1 larva^–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 μg mg lipid^–1 larva^–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.

The relative absorption of fatty acids in brown trout (Salmo trutta) fed a commercial extruded pellet coated with different lipid sources

The objective of the present study was to investigate the fatty acid absorption capabilities of brown trout (Salmo trutta) fed commercial extruded diets. Five commercial extruded pellets, different only in the lipid sources used for fat coating, were tested on juvenile brown trout for 45 days. The trout were reared in fresh water at 14.6 ± 0.4° C and 7.7 ±
0.3 mg/l, temperature and dissolved oxygen, respectively. The tested lipid sources were fish oil, canola oil, oleine oil, swine fat and poultry fat. After the adaptation period faeces were collected by gently stripping from naesthetized fish. Fatty acid analysis was performed on experimental diets and on collected faeces to evaluate the relative absorption capabilities of the trout digestive system with respect to each detected fatty acid. The use of the relative absorption efficiency (rAE) was opted to evaluate the intrinsic capability of each fatty acid to be absorbed. Brown trout showed a
specific preferential order of absorption of the fatty acids, preferring shorter over longer chain fatty acids and preferring the more unsaturated to the more saturated fatty acids. The fatty acid that showed the best relative absorbability was the C18:4n-3 (rAE = 5.14 ± 0.72), which has a fairly short carbon chain, but at the same time a high unsaturation level, followed by the C18:3n-3 (rAE = 3.38 ± 0.30). The fatty acid that showed the worst relative absorbability (rAE = 0.21 ± 0.02) was C24:1n-9.

Effects of dietary lipid sources on flavour volatile compounds of brown trout (Salmo trutta L.) fillet

The high cost and unpredictable availability of fish meal and fish oil (FO) forced feed mill companies to look for alternative ingredients for aquafeeds. In this study, the effects of alternative dietary lipid sources [FO as control, canola oil (CO), oleine oil (OO), poultry fat (PF) and pork lard (PL)] in trout feed on flavour volatile compounds occurring in brown trout (Salmo trutta L.) fillet were evaluated after 70 days of feeding (rearing temperature 14.6°C). Total amounts of volatile compounds identified were higher for fillets of fish fed diets containing only FO as lipid sources. Total amount of alcohols and aldehydes of the fillets were linearly directly related to the percentage content of polyunsaturated fatty acids (PUFA) n-3 of brown trout flesh. The use of alternative dietary lipid sources, modifying the fillet fatty acids composition, affect the total amount of volatile compounds and, changing the relative amount of each volatile compound, affect the flavour of the fish flesh.

Effects of alternative dietary lipid sources on performance, tissues chemical composition, mitochondrial fatty acid oxidation capabilities, and sensory characteristics in brown trout (Salmo trutta L.)

The efficiency of five dietary lipid sources (fish oil as control—C; canola oil—CO; poultry fat—PF; pork lard—PL; and oleine oil—OO) were evaluated in juvenile brown trout (58.4±0.7 g) in an experiment conducted over 70 days at 14.6±0.4 °C. The best growth was observed in fish fed the C diet whereas the PL diet fed fish had the best feed utilization. Significant differences in carcass and muscle proximate composition, but not in liver, were noted among fish fed the different dietary treatments. The fatty acid composition of muscle largely reflected that of the diets, while total cholesterol was not affected. The atherogenicity and the thrombogenicity qualities of the trout flesh were modified by the lipid sources. Sensory analysis did not show any significant differences among the cooked fillets with respect to dietary treatments, while in uncooked products, some significant differences were observed. The carnitine palmitoyltransferase I and II (CPT-I and CPT-II) activities of liver and white muscle were assayed for a better understanding of the potential β-oxidation capability of the different dietary lipid sources. The hepatic, but not white muscle CPT-I and CPT-II activities were affected by dietary treatments. This study showed that alternative lipid sources could be used effectively for oil coating extruded diets for brown trout.

Effect of crude oil extracts from trout offal as a replacement for fish oil in the diets of the Australian native fish Murray cod Maccullochella peelii peelii

The efficacy of trout oil (TO), extracted from trout offal from the aquaculture industry, was evaluated in juvenile Murray cod Maccullochella peelii peelii (25.4-0.81 g) diets in an experiment conducted over 60 days at 23.7-0.8 °C. Five isonitrogenous (48% protein), isolipidic (16%) and isoenergetic (21.8 kJ gm1) diets, in which the fish oil fraction was replaced in increments of 25% (0-100%), were used. The best growth and feed efficiency was observed in fish fed diets containing 50-75% TO. The relationship of specific growth rate (SGR), food conversion ratio (FCR) and protein efficiency ratio (PER) to the amount of TO in the diets was described in each case by second-order polynomial equations (P<0.05), which were: SGR=-0.44TO2+0.52TO+1.23 (r2=0.90, P<0.05); FCR=0.53TO2-0.64TO+1.21 (r2=0.95, P<0.05); and PER=-0.73TO2+0.90TO+1.54 (r2=0.90, P<0.05). Significant differences in carcass and muscle proximate compositions were noted among the different dietary treatments. Less lipid was found in muscle than in carcass. The fatty acids found in highest amounts in Murray cod, irrespective of the dietary treatment, were palmitic acid (16:0), oleic acid (18:1n-9), linoleic acid (18:2n-6) and eicosapentaenoic acid (20:5n-3). The fatty acid composition of the muscle reflected that of the diets. Both the n-6 fatty acid content and the n-3 to n-6 ratio were significantly (P<0.05) related to growth parameters, the relationships being as follows. Percentage of n-6 in diet (X) to SGR and FCR: SGR=-0.12X2+3.96X-32.51 (r2=0.96) and FCR=0.13X2-4.47X+39.39 (r2=0.98); and n-3:n-6 ratio (Z) to SGR, FCR, PER: SGR=-2.02Z2+5.01Z-1.74 (r2=0.88), FCR=2.31Z2-5.70Z+4.54 (r2=0.93) and PER=-3.12Z2-7.56Z+2.80 (r2=0.88) respectively. It is evident from this study that TO could be used effectively in Murray cod diets, and that an n-3:n-6 ratio of 1.2 results in the best growth performance in Murray cod.

Identification and expression of natriuretic peptide receptor Type-A and-B mRNA in freshwater and seawater rainbow trout

Natriuretic peptide receptors mediate the physiological response of  natriuretic peptide hormones. One of the natriuretic peptide receptor types is the particulate guanylyl cyclase receptors, of which there are two identified: NPR-A and NPR-B. In fishes, these have been sequenced and characterized in eels, medaka, and dogfish shark (NPR-B only). The euryhaline rainbow trout provides an opportunity to further pursue examination of the system in teleosts. In this study, partial rainbow trout NPR-A-like and NPR-B-like mRNA sequences were identified via PCR and cloning. The sequence information was used in real-time PCR to examine mRNA expression in a variety of tissues of freshwater rainbow trout and rainbow trout acclimated to 35 parts per thousand seawater for a period of 10 days. In the excretory kidney and posterior intestine, real-time PCR analysis showed greater expression of NPR-B in freshwater fish than in those adapted to seawater; otherwise, there was no difference in the expression of the individual receptors in fresh water or seawater. In general, the expression of the NPR-A and NPR-B type receptors was quite low. These findings indicate that NPR-A and NPR-B mRNA expression is minimally altered under the experimental regime used in this study.