Johanne Dalsgaard

Dietary methionine level affects growth performance and hepatic gene expression of GH–IGF system and protein turnover regulators in rainbow trout (Oncorhynchus mykiss) fed plant protein-based diets

The effects of dietary level of methionine were investigated in juvenile rainbow trout (Oncorhynchus mykiss) fed five plant-based diets containing increasing content of crystalline methionine (Met), in a six week growth trial. Changes in the hepatic expression of genes related to i) the somatotropic axis: including the growth hormone receptor I (GHR-I), insulin-like growth hormones I and II (IGF-I and IGF-II, respectively), and insulin-like growth hormone binding protein-1b (IGFBP-1b); and ii) protein turnover: including the target of rapamycin protein (TOR), proteasome 20 delta (Prot 20D), cathepsin L, calpains 1 and 2 (Capn 1 and Capn 2, respectively), and calpastatin long and short isoforms (CAST-L and CAST-S, respectively) were measured for each dietary treatment. The transcript levels of GHR-I and IGF-I increased linearly with the increase of dietary Met content (P<0.01), reflecting overall growth performances. The apparent capacity for hepatic protein degradation (derived from the gene expression of TOR, Prot 20D, Capn 1, Capn 2, CAST-L and CAST-S) decreased with increasing dietary Met level in a relatively linear manner. Our results suggest that Met availability affects, directly or indirectly, the expression of genes involved in the GH/IGF axis response and protein turnover, which are centrally involved in the regulation of growth.

Glycogenesis and de novo lipid synthesis from dietary starch in juvenile gilthead sea bream (Sparus aurata) quantified with stable isotopes

The effects of replacing a digestible energy source from fat (fish oil) with carbohydrate (wheat starch) on performance, glycogenesis and de novo lipogenesis was examined in triplicate groups of juvenile gilthead sea bream (Sparus aurata), fed four extruded experimental diets. In order to trace the metabolic fate of dietary starch, 0.7% wheat starch was replaced with isotope-labelled starch (>98% 13C). Fish were fed the experimental diets for three consecutive 10 d periods, and isotope ratio MS was applied to quantify 13C enrichment of liver and whole-body glycogen and lipid pools over the three feeding periods. Glycogenesis originating from dietary starch accounted for up to 68.8 and 38.8% of the liver and whole-body glycogen pools, respectively, while up to 16.7% of the liver lipid could be attributed to dietary starch. Between 5 and 8% of dietary starch carbon was recovered in whole-body lipid, and estimated deposition rates of de novo synthesised lipid originating from starch ranged from 18.7 to 123.7 mg/kg biomass per d. Dietary treatments did not significantly affect growth, feed performance or body composition of the fish, while the hepatosomatic index and glycogen content of whole fish and livers correlated directly with dietary starch inclusion level. The study suggests that gilthead sea bream efficiently synthesises glycogen from both dietary starch and endogenous sources. In contrast, lipogenesis from carbon derived from starch seems to play a minor role in overall lipid synthesis and deposition under the specified experimental conditions.

Effects of dietary energy density and digestible protein:energy ratio on de novo lipid synthesis from dietary protein in gilthead sea bream (Sparus aurata) quantified with stable isotopes.

The effects of varying dietary digestible protein (DP) and digestible energy (DE) content on performance, nutrient retention efficiency and the de novo lipogenesis of DP origin were examined in triplicate groups of gilthead sea bream (Sparus aurata), fed nine extruded experimental diets. In order to trace the metabolic fate of dietary protein, 1·8% fishmeal was replaced with isotope-labelled whole protein (.98% 13C). The experiment was divided into a growth period lasting 89 d, growing fish from approximately 140 to 350 g, followed by a 3 d period feeding isotope-enriched diets. Isotope ratio MS was applied to quantify the 13C enrichment of whole-body lipid from dietary DP. Between 18·6 and 22·4% of the carbon derived from protein was recovered in the lipid fraction of the fish, and between 21·6 and 30·3% of the total lipid deposited could be attributed to dietary protein. DP retention was significantly improved by reductions in dietary DP:DE ratio, while the opposite was true for apparent digestible lipid retention. Both overall DE retention and whole-body proximate composition of whole fish were largely unaffected by dietary treatments, while feed conversion ratios were significantly improved with increasing dietary energy density. The present study suggests that gilthead sea bream efficiently utilises dietary nutrients over a wide range of DP:DE ratios and energy densities. In addition, they appear to endeavour a certain body energy status rather than maximising growth, which in the present trial was apparent from inherently high de novo lipogenesis originating from DP.

Increasing levels of dietary crystalline methionine affect plasma methionine profiles, ammonia excretion, and the expression of genes related to the hepatic intermediary metabolism in rainbow trout (Oncorhynchus mykiss)

Strictly carnivorous fish with high requirements for dietary protein, such as rainbow trout (Oncorhynchus mykiss) are interesting models for studying the role of amino acids as key regulators of intermediary metabolism. Methionine is an essential amino acid for rainbow trout, and works as a signalling factor in different metabolic pathways. The study investigated the effect of increasing dietary methionine intake on the intermediary metabolism in the liver of juvenile rainbow trout. For this purpose, five diets were formulated with increasing methionine levels from 0.60 to 1.29% dry matter. The diets were fed in excess for six weeks before three sampling campaigns carried out successively to elucidate (i) the hepatic expression of selected genes involved in lipid, glucose and amino acid metabolism; (ii) the postprandial ammonia excretion; and (iii) the postprandial plasma methionine concentrations. The transcript levels of enzymes involved in lipid metabolism (fatty acid synthase, glucose 6 phosphate dehydrogenase and carnitine palmitoyl transferase 1 a), gluconeogenesis (fructose-1,6-biphosphatase) and amino acid catabolism (alanine amino transferase and glutamate dehydrogenase) were significantly affected by the increase in dietary methionine. Changes in gene expression reflected to some extent the decrease in ammonia excretion (P=0.022) and in the hepatosomatic index (HSI; P<0.001) when dietary methionine increased. Postprandial plasma methionine concentrations correlated positively with the dietary level (P<0.001) at the different sampling points. The study shows that the expression of several genes related to the hepatic intermediary metabolism in rainbow trout responded in a dose-dependent manner to increasing levels of dietary methionine.

Effects of organic plant oils and role of oxidation on nutrient utilization in juvenile rainbow trout (Oncorhynchus mykiss)

Producing organic fish diets requires that the use of both fishmeal and fish oil (FO) be minimized and replaced by sustainable, organic sources. The purpose of the present study was to replace FO with organic oils and evaluate the effects on feed intake, feed conversion ratio (FCR), daily specific growth rate (SGR) and nutrient digestibility in diets in which fishmeal protein was partly substituted by organic plant protein concentrates. It is prohibited to add antioxidants to organic oils, and therefore the effects of force-oxidizing the oils (including FO) on feed intake and nutrient digestibility was furthermore examined. Four organic oils with either a relatively high or low content of polyunsaturated fatty acids were considered: linseed oil, rapeseed oil, sunflower oil and grapeseed oil. Substituting FO with organic oils did not affect feed intake (P > 0.05), FCR or SGR (P > 0.05) despite very different dietary fatty acid profiles. All organic plant oils had a positive effect on apparent lipid digestibility compared with the FO diet (P < 0.05), whereas there were no effects on the apparent digestibility of other macronutrients when compared with the FO diet (P > 0.05). Organic vegetable oils did not undergo auto-oxidation as opposed to the FO, and the FO diet consequently had a significantly negative effect on the apparent lipid digestibility. Feed intake was not affected by oxidation of any oils. In conclusion, the study demonstrated that it is possible to fully substitute FO with plant-based organic oils without negatively affecting nutrient digestibility and growth performance. Furthermore, plant-based organic oils are less likely to oxidize than FOs, prolonging the shelf life of such organic diets.

Nitrate removal from aquaculture effluents using woodchip bioreactors was improved by adding sulfur granules and crushed seashells

This study examined the effects on nitrate removal when adding sulfur granules and crushed seashells to a woodchip bioreactor treating aquaculture effluents. Using a central composite design, the two components were added at three levels (0.000, 0.125 and 0.250 m3/m3 bioreactor volume) to 13 laboratory-scale woodchip bioreactors, and a response surface method was applied to find and model the optimal mixture ratios with respect to reactor performance. Adding 0.125 m3/m3 sulfur granules improved the total N removal rate from 3.27 ± 0.38 to 8.12 ± 0.49 g N/m3/d compared to pure woodchips. Furthermore, the inclusion of crushed seashells together with sulfur granules helped to maintain the pH above 7.4 and prevent a production (i.e., release) of nitrite. According to the modeled response surfaces, a sulfur granule:crushed seashell:woodchip mixture ratio containing about 0.2 m3 sulfur granules and 0.1 m3 crushed seashells per m3 reactor volume would give the best results with respect to high N removal and minimal nitrite release. In conclusion, the study showed that N removal in woodchip bioreactors may be improved by adding sulfur granules and seashells, contributing to the optimization of woodchip performance in treating aquaculture effluents.