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The present study investigates the response of the marine fish cobia, Rachycentron canadum, to stressors as measured by phosphorylation of the α-subunit of the translational initiation factor, eIF2. eIF2α is the target of phosphorylation by a family of kinases that respond to a range of physiological stressors. Phosphorylation of eIF2α inhibits overall protein synthesis, but also facilitates the reprogramming of gene expression to adapt to, and recover from, stress. The deduced coding sequence of cobia eIF2α has 94% identity to both zebrafish (Danio rerio) and human eIF2α sequences with identical phosphorylation and kinase docking sites. Here we use cobia larvae and a cobia cell line derived from muscle (Cm cells) to investigate the response of cobia eIF2α to various stressors. In Cm cells, phosphorylation of eIF2α is increased by nutrient deficiency and endoplasmic reticulum stress (ER stress), consistent with the activation of the eIF2 kinases, GCN2, and PERK. In cobia juveniles, diet and water temperature affect the phosphorylation state of eIF2α. We conclude that evaluation of eIF2α phosphorylation could function as an early marker to evaluate diet, environmental stressors, and disease in cobia and may be of particular use in optimizing conditions for rearing cobia larvae and juveniles. [ABSTRACT FROM AUTHOR]

This study evaluated the effects of seven diets composed of natural feed components (chopped fish, shrimp, and squid) alone or in combination on the liver metabolite profile of juvenile red drum (Sciaenops ocellatus) cultured in a 24-tank recirculating aquaculture system over the course of 12 weeks using Nuclear Magnetic Resonance (NMR)-based metabolomics. Experimental diets included fish (F), shrimp (SH), squid (SQ), fish and shrimp (FSH), fish and squid (FSQ), shrimp and squid (SHSQ), fish, shrimp, and squid (FSHSQ). A commercial fishmeal-based pelleted diet was used as a control. Fish were fed isocalorically. Red drum liver samples were collected at five different time points: T0, before the start of the trial (n = 12), and subsequently every 3 weeks over the course of 12 weeks (T3, T6, T9, T12), with n = 9 fish/diet/time point. Polar liver extracts were analyzed by NMR-based metabolomics. Multivariate statistical analyses (PCA, PLS-DA) revealed that red drum fed the F diet had a distinct liver metabolite profile from fish fed the other diets, with those fed SH, SQ and the combination diets displaying greater similarities in their metabolome. Results show that 19 metabolites changed significantly among the different dietary treatments, including amino acids and amino acid derivatives, quaternary amines and methylamines, carbohydrates and phospholipids. Specifically, γ-butyrobetaine, N-formimino-L-glutamate (FIGLU), sarcosine and beta-alanine were among the most discriminating metabolites. Significant correlations were found between metabolites and six growth performance parameters (final body weight, total length, condition factor, liver weight, hepatosomatic index, and eviscerated weight). Metabolites identified in this study constitute potential candidates for supplementation in fish feeds for aquaculture and optimization of existing formulations. Additionally, we identified a quaternary amine, γ-butyrobetaine as a potential biomarker of shrimp consumption in red drum. These results warrant further investigation and biomarker validation and have the potential for broader applicability outside of the aquaculture field in future investigations in wild red drum populations and potentially other carnivorous marine fishes. [ABSTRACT FROM AUTHOR]

Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity. [ABSTRACT FROM AUTHOR]

Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity., Competing Interests: The authors declare no conflict of interest.