Thanongsak Thanuthong

LC-PUFA biosynthesis in rainbow trout is substrate limited: use of the whole body fatty acid balance method and different 18:3n-3/18:2n-6 ratios.

Five experimental diets with constant total C18 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.

Effects of Dietary α-Linolenic Acid (18:3n-3)/Linoleic Acid (18:2n-6) Ratio on Fatty Acid Metabolism in Murray Cod (Maccullochella peelii peelii)

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.

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.

Effects of dietary vitamin B6 supplementation on fillet fatty acid composition and fatty acid metabolism of rainbow trout fed vegetable oil based diets.

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.