Fatty acid metabolism (desaturation, elongation and β-oxidation) in rainbow trout fed fish oil- or linseed oil-based diets

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 D-6 and D-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.

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.

Short-term food deprivation before a fish oil finishing strategy improves the deposition of n-3 LC-PUFA, but not the washing-out of C18 PUFA in rainbow trout

This study aimed to test the hypothesis that the efficiency of a finishing period can be improved by reducing the initial fat content of fish fillets, by means of a period of food deprivation. Two groups of rainbow trout (Oncorhynchus mykiss) were fed for an 18-week grow-out period on a vegetable oil-based diet (VO) or a fish oil-based diet (FO). VO fed fish were then split into two sub groups: one (VO/FO) was shifted to the FO diet for 8 weeks, whilst the other (UF/FO) was deprived of food (unfed) for 2 weeks and then fed the FO diet for the remaining 6 weeks. The control treatment (FO/FO) was represented by fish continuously fed FO. The subsequent reduction of total fat in the UF/FO treatment was then responsible for a much faster recovery towards a FO-like fatty acid profile, validating the proposed hypothesis. However, the modification of the fatty acid composition of fish fillets during the feed withholding period, coupled with the postponement of the finishing diet, resulted in only minor beneficial effects of this strategy, and the loss of potential weight gain. However, the n-3 LC-PUFA content in UF/VO fish fillets was significantly higher than fish subjected to the VO/FO treatment.

Introgression of domesticated alleles into a wild trout genotype and the impact on seasonal survival in natural lakes

We tested the fitness consequences of introgression of fast-growing domesticated fish into a wild population. Fry from wild and domesticated rainbow trout (Oncorhynchus mykiss) crosses, F1 hybrids, and first- and second-generation backcrosses were released into two natural lakes. Parentage analysis using microsatellite loci facilitated the identification of survivors, so fitness was estimated in nature from the first-feeding stage. Results indicated that under certain conditions, domesticated fish survived at least as well as wild fish within the same environment. Relative growth and survival of the crosses, however, were highly dependent on environment. During the first summer, fastest-growing crosses had the highest survival, but this trend was reversed after one winter and another summer. Although the F1 hybrids showed evidence of outbreeding depression because of the disruption of local adaptation, there was little evidence of outbreeding depression in the backcrosses, and the second-generation backcrosses exhibited a wild-type phenotype. This information is relevant for assessing the multigenerational risk of escaped or released domesticated fish should they successfully interbreed with wild populations and provides information on how to minimize detrimental impacts of a conservation breeding and/or management programme. These data also further understanding of the selection pressures in nature that maintain submaximal rates of growth.

Echium oil provides no benefit over linseed oil for (n-3) long-chain PUFA biosynthesis in rainbow trout

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.

Asymmetric impact of piscivorous birds on size-structured fish populations

Fish are frequently considered the top predator in freshwater food web models despite evidence that predatory birds can impact fish populations. In this study, we quantified bird predation rates on experimental populations of rainbow trout (Oncorhynchus mykiss (Walbaum, 1792)) created by stocking nine small lakes in British Columbia, Canada. Combining estimates of fish mortality with estimated bird predation rates allowed us to partition fish mortality into that due to birds versus cannibalism. Our results indicated that bird predators had significant impacts on age-1 trout populations, but little impact on age-0 trout. Common loons (Gavia immer Brunnich, 1764) were the principle predator among eight predatory bird species present, apparently consuming nearly 50% of all stocked age-1 trout and explaining almost 50% of variation in mortality rates. Age-1 trout mortality did not differ significantly from zero in lakes without loons. Birds consumed a small proportion of age-0 trout, and estimated consumption explained none of the variation in age-0 trout mortality among lakes. We conclude that birds affect fish populations by asymmetric predation on different age (size) classes and can be important top predators that should not be ignored when characterizing freshwater food webs in lakes.

Triploid and diploid rainbow trout do not differ in their stress response to transportation

We examined the neuroendocrine and cellular stress responses of diploid and triploid rainbow trout Oncorhynchus mykiss to transportation. Juvenile diploid and triploid rainbow trout (28 and 26 g/fish average weight, respectively) were stocked at 100 g/L in replicate 70-L tanks and subjected to transportation for an 8-h period. Subsequent levels of plasma cortisol and glucose and of cellular hepatic glutathione (GSH) and heat shock protein 70 (Hsp70) were similar between ploidy groups, indicating that triploid fish respond to transportation in much the same way as diploid fish. A stationary treatment was also included that involved confinement of experimental fish in similar tanks without transport to determine to what extent high-density containment contributed to the stress response in the absence of the noise and vibration of transport. Unexpectedly, fish in the stationary treatment had significantly higher plasma cortisol and glucose levels than the transported fish; however, this might be attributable to a confounding effect of hyperoxia, as oxygen levels fluctuated between 150% and 460% saturation in the stationary tank, while those in the transported tank remained within 100–200% saturation. We suggest that when long stops are necessary while transporting fish, water agitators be used to preclude the additional stress of excessive gas saturation. This may be particularly important for triploid fish, which had lower hepatic GSH levels than diploid fish as well as a low level of mortality in the stationary treatment, unlike the diploid fish.

Mechanisms for climate-induced mortality of fish populations in whole-lake experiments

The effects of climate change on plant and animal populations are widespread and documented for many species in many areas of the world. However, projections of climate impacts will require a better mechanistic understanding of ecological and behavioral responses to climate change and climate variation. For vertebrate animals, there is an absence of whole-system manipulative experiments that express natural variation in predator and prey behaviors. Here we investigate the effect of elevated water temperature on the physiology, behavior, growth, and survival of fish populations in a multiple whole-lake experiment, by using 17 lake-years of data collected over 2 years with differing average temperatures. We found that elevated temperatures in excess of the optimum reduced the scope for growth through reduced maximum consumption and increased metabolism in young rainbow trout, Oncorhynchus mykiss. Increased metabolism at high temperatures resulted in increased feeding activity (consumption) by individuals to compensate and maintain growth rates similar to that observed at cooler (optimum) temperatures. However, greater feeding activity rates resulted in greater vulnerability to predators that reduced survival to only half that of the cooler year. Our work therefore identifies temperature-dependent physiology and compensatory feeding behavior as proximate mechanisms for substantial climate-induced mortality in fish populations at the scale of entire populations and waterbodies.

Density-dependent mortality is mediated by foraging activity for prey fish in whole-lake experiments

1. Whereas the effects of density-dependent growth and survival on population dynamics are well-known, mechanisms that give rise to density dependence in animal populations are not well understood. We tested the hypothesis that the trade-off between growth and mortality rates is mediated by foraging activity and habitat use. Thus, if depletion of food by prey is density-dependent, and leads to greater foraging activity and risky habitat use, then visibility and encounter rates with predators must also increase.

2. We tested this hypothesis by experimentally manipulating the density of young rainbow trout (Oncorhynchus mykiss) at risk of cannibalism, in a replicated single-factor experiment using eight small lakes, during an entire growing season.

3. We found no evidence for density-dependent depletion of daphnid food in the near-shore refuge where most age-0 trout resided. Nonetheless, the proportion of time spent moving by individual age-0 trout, the proportion of individuals continuously active, and use of deeper habitats was greater in high density populations than in low density populations. Differences in food abundance among lakes had no effect on measures of activity or habitat use.

4. Mortality of age-0 trout over the growing season was higher in high density populations, and in lakes with lower daphnid food abundance. Therefore, population-level mortality of age-0 trout is linked to greater activity and use of risky habitats by individuals at high densities. We suspect that food resources were depleted at small spatial and temporal scales not detected by our plankton sampling in the high density treatment, because food-dependent activity and habitat use by age-0 trout occurs in our lakes when food abundance is experimentally manipulated

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.

Population consequences of a predator-induced habitat shift by trout in whole-lake experiments

In a replicated whole-lake experiment, we (a) tested for the existence of a flexible habitat shift in response to predator presence in age-0 rainbow trout (Oncorhynchus mykiss) at risk of cannibalism and (b) evaluated the population-level consequences of habitat shifts in terms of growth and survival over their first growing season. Daphnid food and adult trout predators were substantially more abundant in pelagic than in littoral habitats. Age-0 trout used all habitats in populations without adult trout predators, whereas age-0 trout were observed only in the less profitable littoral habitat in populations with adult trout. Consequently, mean fall mass of age-0 trout in the presence of predators was almost half that observed in populations without adult trout. Despite the shift in habitat use, age-0 trout experienced 90% mortality when adult trout predators were present, in comparison to only 36% mortality when absent. We conclude that the commonly observed habitat shifts by fish at risk of predation, observed at smaller scales, do in fact occur at the whole-system scale over long time intervals. These results suggest that fish are able to perceive risk at large spatial scales and thus take advantage of profitable (but normally risky) habitats when predators are absent, or move to less profitable refuge habitats when predators are present.

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.

Fatty acid-specific alterations in leptin, PPARα, and CPT-1 gene expression in the rainbow trout

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.