Adverse effects of excessive dietary arachidonic acid on survival, PUFA-derived enzymatic and non-enzymatic oxylipins, stress response in rainbow trout fry

Abstract

Arachidonic acid (C20: 4n-6, AA) plays a fundamental role in fish physiology, influencing growth, survival and stress resistance. However, imbalances in dietary AA can have detrimental effects on fish health and performance. Optimal AA requirements for rainbow trout have not been established. This study aimed to elucidate the effects of varying dietary AA levels on survival, growth, long-chain polyunsaturated fatty acid (LC-PUFA) biosynthetic capacity, oxylipin profiles, lipid peroxidation, and stress resistance of rainbow trout fry. Over a period of eight weeks, 4000 female rainbow trout fry at the resorptive stage (0.12 g) from their first feeding were fed diets with varying levels of AA (0.6%, 1.1% or 2.5% of total fatty acids) while survival and growth metrics were closely monitored. The dietary trial was followed by an acute confinement stress test. Notably, while the fatty acid profiles of the fish reflected dietary intake, those fed an AA-0.6% diet showed increased expression of elongase5, highlighting their inherent ability to produce LC-PUFAs from C18 PUFAs and suggesting potential AA or docosapentaenoic acid_n-6 (DPA_n-6) biosynthesis. However, even with this biosynthetic capacity, the trout fed reduced dietary AA had higher mortality rates. The diet had no effect on final weight (3.38 g on average for the three diets). Conversely, increased dietary AA enhanced eicosanoid production from AA, suggesting potential inflammatory and oxidative consequences. This was further evidenced by an increase in non-enzymatic lipid oxidation metabolites, particularly in the AA-2.5% diet group, which had higher levels of phytoprostanes and isoprostanes, markers of cellular oxidative damage. Importantly, the AA-1.1% diet proved to be particularly beneficial for stress resilience. This was evidenced by higher post-stress turnover rates of serotonin and dopamine, neurotransmitters central to the fish's stress response. In conclusion, a dietary AA intake of 1.1% of total fatty acids appears to promote overall resilience in rainbow trout fry.

Keywords: Arachidonic acid; Confinement stress; Dopamine; Oxylipin; Serotonin; Trout.

Figure 1

Overview of experimental design and analyses performed.

Figure 2

Free oxylipins derived from each PUFA cascade (pg/mg of tissue) in fish 6-h post-feeding according to diets. One-way analysis of variance was carried out in order to assess effects of diet. Replicates correspond to individual fish (n = 6 per treatment). Different letters indicate significant differences between groups, which were investigated with a Tukey post hoc test.

Figure 3

Fish concentration of monoaminergic neurotransmitters according to diets after 48-h fasted and confinement stress (pmol/mg of protein) (A) serotonin and metabolite concentrations (5-HT; 5-HIAA and 5-HIAA/5-HT ratio) and (B) dopamine precursor and metabolite concentrations (L-DOPA; HVA and HVA/LDOPA ratio). One-way analysis of variance was carried out in order to assess effects of diet. Replicates correspond to individual fingerlings (n = 8 per treatment). Different letters indicate significant differences between groups, which were investigated with a Tukey post hoc test. 5-HT 5-hydroxytryptamine, 5-HIAA 5-hydroxyindoleacetic acid, HVA Homovanillic acid, L.DOPA L-3,4-dihydroxyphenylalanine. n.s. not significate.