Temperature induced variation in gene expression of thyroid hormone receptors and deiodinases of European eel (Anguilla anguilla) larvae

Thyroid hormones (THs) are key regulators of growth, development, and metabolism in vertebrates and influence early life development of fish. TH is produced in the thyroid gland (or thyroid follicles) mainly as T4 (thyroxine), which is metabolized to T3 (3, 5, 3’-triiodothyronine) and T2 (3, 5-diiodothyronine) by deiodinase (DIO) enzymes in peripheral tissues. The action of these hormones is mostly exerted by binding to a specific nuclear thyroid hormone receptor (THR). In this study, we i) cloned and characterized thr sequences, ii) investigated the expression pattern of the different subtypes of thrs and dios, and iii) studied how temperature affects the expression ofthose genes in artificially produced early life history stages of European eel (Anguilla anguilla), reared in different thermal regimes (16, 18, 20 and 22°C) from hatch until first-feeding. We identified 2 subtypes of thr (thrα and thrβ) with 2 isoforms each (thrαA, thrαB, thrβA, thrβB) and 3 subtypes of deiodinases (dio1, dio2, dio3). All thr genes identified showed high similarity to the closely related Japanese eel (Anguilla japonica). We found that all genes investigated in this study were affected by larval age (in real time or at specific developmental stages), temperature, and/or their interaction. More specifically, the warmer the temperature the earlier the expression response of a specific target gene. In real time, the expression profiles appeared very similar and only shifted with temperature. In developmental time, gene expression of all genes differed across selected developmental stages, such as at hatch, during teeth formation or at first-feeding. Thus, we demonstrate that the expression of thrs and dios show sensitivity to temperature and are involved in and during early life development of European eel

The Korowai Framework: Assessing GE through the Values the ART Confederation Associates with Ngarara

The aim of this thesis is to assess genetic engineering (GE) through the values that the Confederation of Te Ati Awa, Ngati Raukawa ki te tonga and Ngati Toarangatira (the ART Confederation) associates with ngarara. The Korowai Framework was developed to conduct this assessment. Interviews were conducted with 14 participants from across the ART Confederation on the values they associate with ngarara and their interpretations of GE. The values associated with ngarara that were identified in the interviews, were used constitute the kaupapa of the Korowai Framework. The key values identified are: mauri, whakapapa, tohu, tapu, and kaitiakitanga. It emerged from the interviews that ngarara appeal to us to be conscious of our intricately bound connection to and dependency on living systems. The assessment through the Korowai Framework found that the outcomes of GE do not uphold the values associated with ngarara. Participants articulated significant concerns that GE confounds the ART Confederation's control over their relationship with the world around them. This thesis has demonstrated that the Korowai Framework can be used as a tool for the Confederation to get to the decision making table with a comprehensive evidence based understanding of the people's position on GE from which they can negotiate. It demonstrates that robust and legitimate assessment of GE can be conducted using theories, methodologies, kaupapa, tikanga, and frameworks that are specific to the ART Confederation

Effects of the total replacement of fish-based diet with plant-based diet on the hepatic transcriptome of two European sea bass (Dicentrarchus labrax) half-sibfamilies showing different growth rates with the plant-based diet

Background: Efforts towards utilisation of diets without fish meal (FM) or fish oil (FO) in finfish aquaculture have been being made for more than two decades. Metabolic responses to substitution of fishery products have been shown to impact growth performance and immune system of fish as well as their subsequent nutritional value, particularly in marine fish species, which exhibit low capacity for biosynthesis of long-chain poly-unsaturated fatty acids (LC-PUFA). The main objective of the present study was to analyse the effects of a plant-based diet on the hepatic transcriptome of European sea bass (Dicentrarchus labrax). Results: We report the first results obtained using a transcriptomic approach on the liver of two half-sibfamilies of the European sea bass that exhibit similar growth rates when fed a fish-based diet (FD), but significantly different growth rates when fed an all-plant diet (VD). Overall gene expression was analysed using oligo DNA microarrays (GPL9663). Statistical analysis identified 582 unique annotated genes differentially expressed between groups of fish fed the two diets, 199 genes regulated by genetic factors, and 72 genes that exhibited diet-family interactions. The expression of several genes involved in the LC-PUFA and cholesterol biosynthetic pathways was found to be up-regulated in fish fed VD, suggesting a stimulation of the lipogenic pathways. No significant diet-family interaction for the regulation of LC-PUFA biosynthesis pathways could be detected by microarray analysis. This result was in agreement with LC-PUFA profiles, which were found to be similar in the flesh of the two half-sibfamilies. In addition, the combination of our transcriptomic data with an analysis of plasmatic immune parameters revealed a stimulation of complement activity associated with an immunodeficiency in the fish fed VD, and different inflammatory status between the two half-sibfamilies. Biological processes related to protein catabolism, amino acid transaminations, RNA splicing and blood coagulation were also found to be regulated by diet, while the expression of genes involved in protein and ATP synthesis differed between the half-sibfamilies. Conclusions: Overall, the combined gene expression, compositional and biochemical studies demonstrated a large panel of metabolic and physiological effects induced by total substitution of both FM and FO in the diets of European sea bass and revealed physiological characteristics associated with the two half-sibfamilies

Nutritional components affecting skeletal development in fish larvae

International audienc

Combined effects of dietary HUFA level and temperature on sea bass (Dicentrarchus labrax) larvae development

International audienceThe purpose of this study was to investigate the combined effect of the incorporation of vegetable products in diet and temperature on enzymatic pathways for high unsaturated fatty acids (HUFA) desaturation in sea bass larvae. Four replicated groups were fed a low (LH; 0.8% EPA + DHA) or a high (HH; 2.2% EPA + DHA) n-3 HUFA microparticulated diet from mouth opening, six days post-hatching and were reared at 16 or 22 °C. The four experimental conditions (LH16, HH16, LH22 and HH22) were tested for 45 days. At the end of the experiment, body weight, total length and biomass were affected by temperature (P b 0.001), while biomass as well as fresh body weight was also influenced by diet (P b 0.05 and P b 0.001 respectively). This always lead to the same ranking of experimental conditions: HH22 N LH22 N HH16 N LH16. The larval skeletal development was more advanced in 22 °C-groups than in 16 °C-ones (P b 0.001), while it was not affected by diet. Amylase and trypsin pancreatic secretions did not vary between d-25 and d-45, indicating that pancreatic maturation was achieved at d-25. Low temperature combined with low dietary HUFA delayed intestinal maturation (P b 0.001), while low temperature combined with high HUFA diet allowed larvae compensating for the initial intestinal maturation retardation. Lipase gene expression was down-regulated in HH16 group at d-25 (P b 0.05) and in the two 16 °C-groups at d-45 (P b 0.001), while lipase enzymatic activity was similar in all groups. This suggested the presence of a post-transcriptional regulation of this gene. PPAR α and PPAR β were not affected neither by temperature, nor by diet, suggesting that lipid metabolism was not significantly affected by a lowering in dietary n-3 HUFA when isolipidic diets were used. A higher DHA content was found in larvae than in their diets (× 2 for LH; ×1.5 for HH) but the DHA content in PL of d-45 LH larvae was lower than the initial one, which revealed a HUFA deficiency in this group. Delta 6-desaturase (Δ6D) gene expression was significantly up-regulated by HUFA deprived diet (P b 0.05) whatever the temperature was. This was supported by the increase in 18:3n-6 in LH larvae (P b 0.001), which indicated a desaturation from 18:2n-6 by the Δ6D. This study clearly showed that larvae were able to adapt to an n-3 deprived diet by a stimulation of enzymatic pathways for HUFA desaturation, and that this adaptation was not affected by temperature

Is it possible to influence European sea bass (Dicentrarchus labrax) juvenile metabolism by a nutritional conditioning during larval stage?

International audienceThe purpose of this study was to check if it is possible to influence sea bass juvenile metabolism by a conditioning of larvae from day 6 post hatching to day 45 to a low or a high HUFA compound diet (LH, 0.8% EPA + DHA and HH, 2.2% EPA + DHA) when reared at 16 or 22 °C. Following a 3-month intermediate period (at 19 °C using a commercial diet), the adaptability of the 4 initial larval groups to a HUFA experimental deprived diet (0.5% EPA + DHA) were tested at 19 °C in a 60 day-experiment (d-151– 211). The four experimental duplicated conditions were ex-LH16 and ex-HH16 for the 2 groups previously reared at 16 °C (initial weight, 7.3 ± 0.5 g) and ex-LH22 and ex-HH22 for the 2 groups previously reared at 22 °C (initial weight, 11.1 ± 0.5 g). Survival was maximal and there was a 1.6–2 fold increase in mass during the experiment. Growth was similar in the 4 experimental groups: NS difference in growth curve slopes (P = 0.7). At the end of the experiment (d-211), whole body fat levels were in the same range in all groups (13–15% WW). The fatty acid (FA) composition in polar lipids (PL) and total lipids (TL) were significantly affected by initial weight related to larvae conditioning, which can be mainly attributed to a dilution effect (impact of initial FA content on final FA content versus relative mass increase during the course of the experiment). Conversely to this trend, DHA content in PL was higher in the ex-LH groups than in the ex-HH groups whatever thermal conditioning of larvae was. This indicated that ex-LH groups had a better capacity to adapt to a deficient HUFA diet than ex-HH fish. The relative expression of the delta-6 desaturase (Δ6D) was significantly higher in ex-LH than in ex-HH groups (P b 0.001) between d-151 and d-181, which suggested that Δ6D transcription in ex-LH groups was positively modulated by the HUFA-deprived diet. This stimulation of the first step of the desaturation/elongation pathway could allow synthesizing FA needed to compensate low dietary HUFA supply. This study shows for the first time that it seems possible to influence juvenile fish metabolism by a nutritional conditioning during the larval stage