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On the compatibility of fish meal replacements in aquafeeds for rainbow trout. A combined metabolomic, proteomic and histological study.

ABSTRACT: 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.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Oncorhynchus Mykiss (rainbow Trout) (salmo Gairdneri)

TISSUE(S): Liver

SUBMITTER: Antonio Palomba

LAB HEAD: Salvatore Pisanu

PROVIDER: PXD033201 | Pride | 2022-07-18

REPOSITORIES: Pride

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Project description:Sustainable aquaculture, which entails proportional replacement of fish-based feed sources by plant-based ingredients, is impeded by the poor growth response frequently seen in fish fed high levels of plant ingredients. To evaluate the potential to improve, by means of early nutritional exposure, the growth of fish fed plant-based feed, rainbow trout swim-up fry were fed for 3 weeks either a plant-based diet (diet V, V-fish) or a diet containing fishmeal and fish oil as protein and fat source (diet M, M-fish). After this 3-week nutritional history period, all V- or M-fish received diet M for a 7-month intermediate growth phase. Both groups were then challenged by feeding diet V for 25 days during which voluntary feed intake, growth, and nutrient utilization were monitored (V-challenge). The results of the V-challenge showed a higher growth rate and higher feed intake in fish of nutritional history V compared to M (see PMID:24386155). To identify the molecular mechanisms that govern the enhancement of feed acceptance following early-feed exposure of diet V, we performed microarray expression analysis using swim-up fry collected at the end of early-feeding exposure (diet M and diet V), as well as brain and liver of juveniles sampled at the end of the V-challenge (M-his and V-his), and employing a rainbow trout microarray platform with 8 X 60K probes per slide. Persistently differentially expressed genes were identified by isolating probes that were differentially expressed (fold change ≥ 1.5 and p {nutritional history} ≤ 0.05), both at the end of the early exposure, and in either brain or liver of juvenile trout at the end of the V-challenge. Overall, 1787 (3-week + Brain) and 924 (3-week + Liver) mRNA probes were found to be persistently affected by nutritional history of early-feeding exposure. This positive response is encouraging as a potential strategy to improve the use of plant-based feed in fish, of interest in the field of fish farming and animal nutrition in general. Furthermore, the genes associated with mediating this positive early feeding effect, could serve as potential biomarkers to evaluate increasing acceptability of plant-based diets in fish farming.

Project description:We have constructed a rainbow trout high-density oligonucleotide microarray by using all the available tentative consensus (TC) sequences from the Rainbow Trout Gene Index database (The Computational Biology and Functional Genomics Lab., Dana Farber Cancer Institute and Harvard School of Public Health). The Rainbow Trout Gene Index integrates research data from all available international rainbow trout genomic research projects. The newly designed microarray incorporates 37,394 unique transcript-specific oligonucleotide probes, 60-mer long each. The microarray was printed according to our design by Agilent Technologies using the 4 X 44-design format and contains 1417 Agilent control spots. The performance of the new microarray platform was evaluated by analyzing gene expression associated with the rainbow trout vitellogenesis-induced muscle atrophy. These chips can be ordered from Agilent using design number 016320. This microarray is anticipated to open new avenues of research that will aid in the development of novel strategies to enhance growth efficiency and quality in salmonid species. Keywords: Development of an oligo-array for rainbow trout The performance of the new microarray platform was evaluated by analyzing transcriptome response associated with the rainbow trout vitellogenesis-induced muscle atrophy. Severe muscle deterioration accompanies the physiological responses of the energetic demands of the rainbow trout spawning/vitellogenesis. Atrophying muscle of fertile fish had 11% less extractable muscle (g/bw) and 11% less protein content compared to non-atrophying muscle of sterile fish (p<0.01). The rainbow trout was used to profile changes in gene expression of atrophying muscles. Gene expression levels were determined by comparing the amount of mRNA transcript present in the experimental sample (fertile fish) to the control (sterile fish). RNAs isolated from each experimental fish were run on separate microarrays in independent experiments, with no pooling. A total of 8 fish were used in the microarray experiments (4 replicates x 2 groups). Fluorophors (Cy3 and Cy5) were randomly assigned to RNA from each of the atrophying and nonatrophying muscles to limit the dye effect.

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On the compatibility of fish meal replacements in aquafeeds for rainbow trout. A combined metabolomic, proteomic and histological study.

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