Project description:The sustainable development of modern aquaculture must rely on a significant reduction of the fish meal (FM) used in aquafeed formulations. However, FM substitution with alternative ingredients in diets for carnivorous fish species often showed reduced nutrient absorption, significantly perturbed metabolisms and histological changes at both hepatic and intestinal level. In the present study, adult rainbow trout (Oncorhynchus mykiss) were fed three different experimental aquafeed formulations. A control diet with higher FM content (27.3%) than two test formulations in which fish meal was substituted with two more sustainable and promising alternatives: insect meal (Hermetia illucens larvae=10.1%, FM=11.6%) and poultry by-products meal (PBM=14.8%; FM=11.7%). Combined metabolomics and proteomics analyses of fish liver, together with histological examination of liver and intestine demonstrated that a well balanced formulation of nutrients in the three diets allowed high metabolic compatibility of either substitutions, paving the way for innovative and sustainable use of novel raw materials for the fish feed industry. Results show that the main metabolic pathways of nutrient absorption and catabolism were essentially unaltered by alternative feed ingredients, and also histological alterations were negligible. It is demonstrated that substitution of fish meal with sustainable alternatives does not impact on fish metabolism, given proper efforts are put in fulfilling nutritional requirements of rainbow trout.
Project description:<p>Dietary glycerol supplementation in aquaculture feed is seen as an alternative and inexpensive way to fuel fish metabolism, attenuate metabolic utilization of dietary proteins and, subsequently reduce nitrogen excretion. In this study, we evaluated the impact of dietary glycerol supplementation has on nitrogen excretion of European seabass (Dicentrarchus labrax) and its effects on metabolite profile and bacterial community composition of gut digesta. These effects were evaluated in a 60-day trial with fish fed diets supplemented with 0, 2.5 or 5% (w/w) refined glycerol. Nuclear magnetic resonance spectroscopy and high-throughput 16S rRNA sequencing characterized the effects of glycerol supplementation of digesta metabolite and bacterial community composition of 6h postprandial fish. Our results showed ammonia excretion was not altered by dietary glycerol supplementation and the highest glycerol dosage was associated with significant increases in amino acids and a significant decrease of ergogenic creatine in digesta metabolome. Concomitantly, significant decreases in putative amino acid degradation pathways were detected in the predicted metagenome analysis, suggesting a metabolic shift. Taxon-specific analysis revealed significant increases in abundance of some specific genera (e.g. Burkholderia and Vibrio) and bacterial diversity. Overall, our results indicate glycerol supplementation may decrease amino acid catabolism without adversely affecting fish gut bacterial communities.</p>
Project description:Responsible aquaculture growth requires practices adjusted to higher knowledge based industrial standards that can secure good fish welfare, health, and low environmental impacts, alongside with the production of safe and nutritious consumer products and the economic sustainability of the business. The fish feed ingredient pallet is ever changing, from marine to plant based, and recently evolving to incorporate increasing amounts of low trophic, side stream and circular economy based raw materials, each one contributing with variable amounts and qualities of macro- and micronutrients. Meeting the micronutrient requirement of farmed fish for healthy and efficient growth under normal and challenging conditions is of paramount importance. Trace minerals are essential for critical life processes, including respiration, protein and lipid metabolism, immune responses, and cell REDOX balance. The study results bring new knowledge on Atlantic salmon physiology, growth, welfare status and utilization efficiency of trace minerals of different origin and dietary inclusion levels revealing differential effects on fillet yield, fillet technical and nutritional quality, bone strength, skin morphology, organ mineralization, midgut transcriptome, trace mineral apparent digestibility and retention efficiency, and growth.
Project description:<p>Aquafeed is one of the main costs in aquaculture and major efforts are being made to find sustainable ingredients that could improve production sustainability and efficiency. Our research focused on the inclusion of glycerol in the feed of European seabass (Dicentrarchus labrax), as it is a cheap carbon substrate expected to enter metabo- lism through the glycolytic pathway. The Nuclear Magnetic Resonance (NMR) spectroscopy provides accurate and high-throughput metabolic profiles, allowing quantification and evaluation of the effects of experimental diets in the context of fish nutrition. The aim of the present study was to assess the applicability of the automated on-line profiling routine Bayesil to fish blood plasma and compare with the manual targeted metabolite profiling approach. After a 60-day feeding trial (0%, 2.5%, 5.0% glycerol supplemented diets), fish were sacrificed at 6 h and 24 h after last meal and blood was sampled. Filtered plasma samples were analysed by NMR, followed by two metabolite-profiling approaches: i) an automated spectral profiling provided by the Bayesil platform, and ii) a manual metabolite profiling. Spectra were analysed with the automated Bayesil routine, identifying and quantifying 51 metabolites, in a reproducible, fast and user-independent method. On the other hand, the manual approach only managed to quantify 20 metabolites. However, under supraphysiological concentrations of glycerol, namely in the 5.0% diet, the Bayesil algorithm was not able to properly integrate the glycerol and glycine signals. The Bland-Altman and PLS-DA analysis revealed differences; nonetheless, with the exception of acetic acid, creatinine, formate, glutamine and threonine, all other metabolites presented a low bias and/or a strong correlation coefficient. Overall, it is expected a reasonable tradeoff between the accuracy in metabolite quantification and the increase of variables.The automated Bayesil database may be applied as a useful tool for systematic monitoring of fish plasma profile in aquaculture research and industrial quality control analysis, to generate robust variance-based multivariate statistical models. To our knowledge, this is the first time a NMR-based high-throughput automated routine is evaluated in aquaculture research.</p>
Project description:Screening has revealed that modern-day feeds used in Atlantic salmon aquaculture might contain trace amounts of agricultural pesticides. To reach slaughter size, salmon are produced in open net pens in the sea. Unconsumed feed pellets and undigested feces deposited beneath the net pens represent a source of contamination for marine organisms. To examine the impacts of long-term and continuous dietary exposure to an organophosphorus pesticide (OP) found in Atlantic salmon feed, we fed juvenile Atlantic cod (Gadus morhua), an abundant species around North Atlantic fish farms, three concentrations (0.5, 4.2 and 23.2 mg/kg) of chlorpyrifos-methyl (CPM) for 30 days. Endpoints included liver and bile bioaccumulation, liver transcriptomics and metabolomics, as well as plasma cholinesterase activity, cortisol, liver 7-ethoxyresor-ufin-O-deethylase (EROD) activity and hypoxia tolerance. The results show that Atlantic cod can accumulate relatively high levels of CPM in the liver after continuous exposure, which is then metabolized and excreted via the bile. All three exposure concentrations led to significant inhibition of plasma cholinesterase activity, the primary target of CPM. Transcriptomics profiling pointed to effects on cholesterol and steroid biosynthesis. Metabolite profiling revealed that CPM induced responses reflecting detoxification by glutathione-S-transferase, inhibition of monoacylglycerol lipase, potential inhibition of carboxylesterase, and increased demand for ATP, followed by secondary inflammatory responses. A gradual hypoxia challenge test showed that all groups of exposed fish were less tolerant to low oxygen saturation than the controls. In conclusion, this study suggests that wild fish continuously feeding on leftover pellets near fish farms over time may be vulnerable to OPs.
Project description:There is an increasing drive to replace fish oil (FO) in finfish aquaculture diets with vegetable oils (VO), driven by the short supply of FO derived from wild fish stocks. Little is known of the consequences for fish health after such substitution. The effect of dietary VO on hepatic gene expression was determined in Atlantic salmon (Salmo salar) byg a cDNA microarray analysis. Post-smolt farmed salmon were reared for x weeks on diets where the FO component of the feed was replaced with one of three different VOs - rapeseed (RO), soybean (SO) or linseed (LO). RNA from five fish fed on each diet was extracted. A total of 20 cDNA microarray hybridisations - TRAITS / SGP Atlantic salmon 17k feature cDNA microarray - were performed - 4 diets (three VO + FO control) x 5 individuals - using a common pooled reference control design. Data were obtained from 19 of the 20 hybridisations.
Project description:Cyberlindnera jadinii yeast is a potential sustainable novel feed ingredient for aquaculture industries. Yeasts contain bio-active components and proteins such as beta-glucans, mannans, nucleic acids and proteins that can enhance fish immunity against the disease. In our study, we focused on the characterization of intestinal immunoregulatory pathways in zebrafish (Danio rerio) by quantifying the intestine proteins with isobaric tags for relative and absolute quantitation (iTRAQ) and 2D LC-MS/MS approach. Zebrafish were fed either a control diet (C) or a diet supplemented with autolyzed C. jadinii (ACJ). The KEGG pathways analysis revealed that compared with the control diet, the ACJ yeast diet induced an increased abundance of proteins related to arginine and proline metabolism, phagosome, C-lectin receptor signalling pathway, ribosome pathway and PPAR signalling pathway, which can modulate and enhance the innate response of zebrafish. Moreover, fish fed ACJ yeast diet also showed decreased abundance of proteins associated with inflammatory pathways including apoptosis, necroptosis and ferroptosis pathways. These findings support a mobilization of the innate immune response and a control of inflammatory-related pathways in the intestine of zebrafish. Our findings in the well annotated proteome of zebrafish enabled a detailed investigation of intestinal responses and provide insight into the health-beneficial effects of the yeast species C. jadinii relevant for aquaculture species.
Project description:The sustainable growth of fish aquaculture will require the procurement of non-marine feed sources. Glycerol is a potential feed supplement whose metabolism may spare the catabolism of dietary amino acids, thereby extending the use of the feed protein to other physiological functions such as growth. In the present study, the effects of dietary glycerol supplementation on the muscle and liver metabolomes of rainbow trout (Oncorhynchus mykiss) and European seabass (Dicentrarchus labrax) were evaluated. Fish juveniles were fed diets with 0%, 2.5%, and 5% glycerol. Muscle and liver aqueous fractions were extracted and 1H NMR spectra were acquired. Metabolite profiles derived from the 1H NMR signals were assessed using univariate and multivariate statistical analyses. The adenylate energy charge was determined in the muscle. For both species, the muscle metabolite profile showed more variability compared to that of the liver and was most perturbed by the 5.0% glycerol diet. For the liver metabolite profile, rainbow trout showed fewer differences compared to European seabass. No differences were observed in energy charge between experimental groups for either species. Thus, rainbow trout appeared to be less susceptible to tissue metabolite perturbations, compared to seabass, when the diet was supplemented with up to 5% glycerol.