Effect of an organic acid blend in Nile tilapia growth performance, immunity, gut microbiota, and resistance to challenge against francisellosis.

Organic acids (OAs) are a class of feed additives that have prophylactic and inhibitory properties against pathogenic bacteria. In this study, we investigated growth performance, innate immune response, gut microbiota, and disease resistance against Francisella orientalis F1 in Nile tilapia (Oreochromis niloticus) fed different doses of Bacti-nil®Aqua, a blend of short- and medium-chain OAs. For 21 days, 680 juvenile tilapias were fed a control diet or diets supplemented with a 0.3% (D3) or 0.5% (D5) OA blend. The feed conversion rate of fish fed the 0.5% enriched diet was considerably lower (p < 0.05) than that of the fish fed the basal diet. Lysozyme and serum bactericidal activities were significantly elevated following OA administration. After infection, no differences in the diversity and composition of gut microbiota were observed between the groups. After the bacterial challenge, the mortality was significantly lower in group D5 (p < 0.01). The diet supplemented with Bacti-nil®Aqua (Adisseo) improved the immune response and resistance of tilapia juveniles against F. orientalis infection. Thus, this OA blend could serve as a feed additive with good activity against F. orientalis.

Physiological Benefits of Dietary Lysophospholipid Supplementation in a Marine Fish Model: Deep Analyses of Modes of Action.

Given the hydrophilic structure of lysophospholipids (LPLs), their dietary inclusion translates into a better emulsifying capacity of the dietary components. The present study aimed to understand the mechanisms underlying the growth-promoting effect of LPL supplementation by undertaking deep analyses of the proximal intestine and liver interactomes. The Atlantic salmon (Salmo salar) was selected as the main aquaculture species model. The animals were divided into two groups: one was fed a control diet (C-diet) and the other a feed (LPL-diet) supplemented with an LPL-based digestive enhancer (0.1% AQUALYSO®, Adisseo). The LPL-diet had a positive effect on the fish by increasing the final weight by 5% and reducing total serum lipids, mainly due to a decrease in the plasma phospholipid (p < 0.05). In the intestine, the upregulated interactome suggests a more robust digestive capacity, improving vesicle-trafficking-related proteins, complex sugar hydrolysis, and lipid metabolism. In the liver, the LPL-diet promotes better nutrients, increasing several metabolic pathways. The downregulation of the responses to stress and stimuli could be related to a reduced proinflammatory state. This study on the benefits and modes of action of dietary LPLs opens a new window into fish nutrition and could be extended to other productive species.

Liver proteome response of pre-harvest Atlantic salmon following exposure to elevated temperature.

BACKGROUND: Atlantic salmon production in Tasmania (Southern Australia) occurs near the upper limits of the species thermal tolerance. Summer water temperatures can average over 19 °C over several weeks and have negative effects on performance and health. Liver tissue exerts important metabolic functions in thermal adaptation. With the aim of identifying mechanisms underlying liver plasticity in response to chronic elevated temperature in Atlantic salmon, label-free shotgun proteomics was used to explore quantitative protein changes after 43 days of exposure to elevated temperature. RESULTS: A total of 276 proteins were differentially (adjusted p-value < 0.05) expressed between the control (15 °C) and elevated (21 °C) temperature treatments. As identified by Ingenuity Pathway Analysis (IPA), transcription and translation mechanisms, protein degradation via the proteasome, and cytoskeletal components were down-regulated at elevated temperature. In contrast, an up-regulated response was identified for NRF2-mediated oxidative stress, endoplasmic reticulum stress, and amino acid degradation. The proteome response was paralleled by reduced fish condition factor and hepato-somatic index at elevated temperature. CONCLUSIONS: The present study provides new evidence of the interplay among different cellular machineries in a scenario of heat-induced energy deficit and oxidative stress, and refines present understanding of how Atlantic salmon cope with chronic exposure to temperature near the upper limits of thermal tolerance.

Triploid Atlantic salmon shows similar performance, fatty acid composition and proteome response to diploids during early freshwater rearing.

There is currently renewed interest in farming triploid Atlantic salmon. Improving farming requires identifying triploid specific phenotypic and physiological traits that are uniquely derived from ploidy per se and developed under optimal growing conditions. This study investigated firstly, the impact of ploidy on growth performance and whole body composition of Atlantic salmon at different early freshwater stages [34dph (days post-hatching) alevin, 109dph fry, and 162dph parr] and secondly, whether phenotypic differences at these stages were reflected in protein samples collected from whole fish, white muscle or liver tissue. Female diploid and triploid Atlantic salmon (n=3) were first fed at 35dph and then maintained by feeding to satiation on commercial feeds. Triploids were significantly lower in weight at the late alevin and fry stages but matched diploid weight at the parr stage. The whole-body lipid content was significantly higher for triploids at the parr stage, while the whole-body lipid class profile was broadly similar and was largely not affected by ploidy. Comparative label-free shotgun proteomic analysis did not detect significant alterations in protein expression between diploids and triploids at any growth stage. The present results indicate that ploidy under optimal growing conditions and during early freshwater stages only result in small phenotypic differences in weight and whole body lipid content that were not reflected at the proteome level. These findings suggest that optimal husbandry conditions for freshwater Atlantic salmon are similar between ploidies, at least for all-female populations.

Preliminary Validation of a High Docosahexaenoic Acid (DHA) and α-Linolenic Acid (ALA) Dietary Oil Blend: Tissue Fatty Acid Composition and Liver Proteome Response in Atlantic Salmon (Salmo salar) Smolts.

Marine oils are important to human nutrition as the major source of docosahexaenoic acid (DHA), a key omega-3 long-chain (≥C20) polyunsaturated fatty acid (n-3 LC-PUFA) that is low or lacking in terrestrial plant or animal oils. The inclusion of fish oil as main source of n-3 LC-PUFA in aquafeeds is mostly limited by the increasing price and decreasing availability. Fish oil replacement with cheaper terrestrial plant and animal oils has considerably reduced the content of n-3 LC-PUFA in flesh of farmed Atlantic salmon. Novel DHA-enriched oils with high alpha-linolenic acid (ALA) content will be available from transgenic oilseeds plants in the near future as an alternative for dietary fish oil replacement in aquafeeds. As a preliminary validation, we formulated an oil blend (TOFX) with high DHA and ALA content using tuna oil (TO) high in DHA and the flaxseed oil (FX) high in ALA, and assessed its ability to achieve fish oil-like n-3 LC-PUFA tissue composition in Atlantic salmon smolts. We applied proteomics as an exploratory approach to understand the effects of nutritional changes on the fish liver. Comparisons were made between fish fed a fish oil-based diet (FO) and a commercial-like oil blend diet (fish oil + poultry oil, FOPO) over 89 days. Growth and feed efficiency ratio were lower on the TOFX diet. Fish muscle concentration of n-3 LC-PUFA was significantly higher for TOFX than for FOPO fish, but not higher than for FO fish, while retention efficiency of n-3 LC-PUFA was promoted by TOFX relative to FO. Proteomics analysis revealed an oxidative stress response indicative of the main adaptive physiological mechanism in TOFX fish. While specific dietary fatty acid concentrations and balances and antioxidant supplementation may need further attention, the use of an oil with a high content of DHA and ALA can enhance tissue deposition of n-3 LC-PUFA in relation to a commercially used oil blend.

Sequential protein extraction as an efficient method for improved proteome coverage in larvae of Atlantic salmon (Salmo salar).

Understanding diet- and environmentally induced physiological changes in fish larvae is a major goal for the aquaculture industry. Proteomic analysis of whole fish larvae comprising multiple tissues offers considerable potential but is challenging due to the very large dynamic range of protein abundance. To extend the coverage of the larval phase of the Atlantic salmon (Salmo salar) proteome, we applied a two-step sequential extraction (SE) method, based on differential protein solubility, using a nondenaturing buffer containing 150 mM NaCl followed by a denaturing buffer containing 7 M urea and 2 M thiourea. Extracts prepared using SE and one-step direct extraction were characterized via label-free shotgun proteomics using nanoLC-MS/MS (LTQ-Orbitrap). SE partitioned the proteins into two fractions of approximately equal amounts, but with very distinct protein composition, leading to identification of ∼40% more proteins than direct extraction. This fractionation strategy enabled the most detailed characterization of the salmon larval proteome to date and provides a platform for greater understanding of physiological changes in whole fish larvae. The MS data are available via the ProteomeXchange Consortium PRIDE partner repository, dataset PXD003366.

Modelling the metabolic characteristics of proteins in dairy cattle from co-products of bioethanol processing: comparison of the NRC 2001 model with the DVE/OEB system.

BACKGROUND: Co-products from bioethanol processing include wheat dried distillers grains with solubles (DDGS), corn DDGS, blend DDGS (e.g. wheat/corn at 70:30, 60:40 or 50:50 w/w), triticale DDGS, barley DDGS and pea DDGS. The objective of this study was to compare two systems, the DVE/OEB system versus the NRC 2001 model, in modelling the metabolic characteristics of proteins in dairy cattle from different types of co-products (DDGS) from different bioethanol processing plants. RESULTS: The predicted values from the NRC 2001 model were 10% higher (P < 0.05) in truly absorbable rumen-synthesised microbial protein in the small intestine, 10% lower (P < 0.05) in truly absorbed rumen-undegraded feed protein in the small intestine, 30% lower (P < 0.05) in endogenous protein and 2% lower (P < 0.05) in total truly absorbed protein in the small intestine than the predicted values from the DVE/OEB system. However, no significant difference was detected in terms of the degraded protein balance between the two models (P > 0.05). CONCLUSION: The sensitivity of the two models in detecting differences among DDGS types and between bioethanol plants was similar. The two models coincided in the superior protein value of blend DDGS as well as in the more optimal degraded protein balance (DPB) for corn DDGS. Although the differences between the DVE/OEB system and the NRC 2001 model were significant (P < 0.05) for most outputs owing to differences in some of the concepts and factors used in modelling, the correlations between total truly absorbed protein (DVE) and metabolisable protein (MP) values and between degraded protein balances (DPB(OEB) vs DPB(NRC) ) were also significant (P < 0.05).

Relationship of protein molecular structure to metabolisable proteins in different types of dried distillers grains with solubles: a novel approach.

To date, there has been no study of protein molecular structures affected by bioethanol processing in relation to protein nutritive values of the new co-products of bioethanol production. The objective of the present study was to investigate the relationship between protein molecular structures (in terms of protein α-helix and ß-sheet spectral intensity and their ratio and amide I to amide II spectral intensity and their ratio) and protein rumen degradation kinetics (rate and extent), estimated protein intestinal digestibility and total truly absorbed protein in small intestine (metabolisable protein) in different types of dried distillers grains with solubles (DDGS), such as wheat DDGS, maize DDGS and blend DDGS (wheat:maize = 70:30). The protein molecular structures of the different types of DDGS affected by processing were identified using diffuse reflectance IR Fourier transform spectroscopy. The results showed that the protein structure α-helix to ß-sheet ratio in the DDGS had a strongly negative correlation with estimated intestinal digestibility of ruminally undegraded protein (%dRUP, R - 0.95, P = 0.04), tended to have a significant correlation with the protein PC subfraction (which was undegradable and contained proteins associated with lignin and tannins and heat-damaged proteins) (R 0.91, P = 0.09) and had no correlation (P>0.10) with rumen degradation kinetics (rate and extent), total intestinally absorbed protein supply and degraded protein balance. However, the protein amide I to amide II ratio in the DDGS had a strongly positive correlation with soluble crude protein (CP) (R 0.99, P < 0.01), protein PA subfraction (which was instantaneously solubilised at time zero) (R 0.99, P < 0.01), protein PB2 subfraction (which was intermediately degradable) (R - 0.95, P = 0.04) and total digestible CP (R 0.95, P = 0.04). The amide I to amide II ratio also had strongly negative correlations with ruminally undegraded protein (%RUP: R - 0.96, P = 0.03) and the degraded protein balance (OEB: R - 0.97, P = 0.02), but had no correlation (P>0.10) with the total intestinally absorbed protein supply. Multiple regression results show that the protein structure α-helix to ß-sheet ratio was a better predictor of %dRUP with R² 0.92. The amide I to II ratio was a better predictor of the degraded protein balance with R² 0.93 in the DDGS. In conclusion, the changes in the protein molecular structure α-helix to ß-sheet ratio and the amide I to amide II ratio during bioethanol processing (either due to fermentation processing or due to heat drying) were highly associated with estimated protein intestinal digestibility and degraded protein balance, but were not associated with total intestinally absorbed protein supply from the DDGS to dairy cattle. The present study indicates that a potential novel method could be developed based on the protein molecular structure parameters to improve the estimation of protein value after a validation in a large-scale in vivo study is done.

Estimation of ruminal and intestinal digestion profiles, hourly effective degradation ratio and potential N to energy synchronization of co-products from bioethanol processing.

BACKGROUND: Little research has been conducted to determine the magnitude of the differences in nutritive value among wheat dried distillers' grains with solubles (DDGS), corn DDGS and blend DDGS, or between different bioethanol plants. The objectives of this study were to compare different types of DDGS and different bioethanol plants in terms of: (1) rumen degradation kinetics profile of each DDGS component and rumen availability; (2) intestinal digestion profile of rumen undegraded protein; (3) hourly effective rumen degradation ratio and potential N-to-energy synchronization; (4) the role of acid detergent insoluble nitrogen in the determination of nutrient availability of DDGS. In addition, these parameters were compared in DDGS as opposed to parental grain. RESULTS: (1) The effective degradability of dry matter in DDGS samples increased as the content of feedstock wheat increased. DDGS are a good source of rumen-undegradable protein. The protein content of DDGS derived from wheat is higher relative to that derived from corn; however, the undegradability of the protein fraction increases as the proportion of corn in the feedstock augments. (2) In addition, DDGS provide significant amounts of rumen-degradable protein, which increased as the content of wheat in the feedstock increased. This indicates a potential loss of N when high levels of DDGS are included in the diet. (3) Acid detergent insoluble crude protein (ADICP) levels were low across DDGS samples, revealing no effect on ruminal and intestinal disappearance of protein. However, consideration should be given to the numerical differences in digestibility of rumen-undegradable protein and the relation to ADICP content. (4) Further research with a higher number of samples and higher variability in the ADICP content should be undertaken to investigate the effect of ADICP on rumen and intestinal disappearance of DDGS protein. CONCLUSION: The digestive characteristics of each DDGS component (dry matter, organic matter, crude protein and neutral detergent fiber), the hourly effective degradation ratio between N and organic matter, and the intestinal availability of rumen-undegradable protein differed significantly (P < 0.05) among wheat DDGS, blend DDGS and corn DDGS, and to a lesser extent between the different bioethanol plants. These results indicate that it is inappropriate to assume fixed rumen and intestinal degradation characteristics for DDGS without considering factors such as DDGS type and bioethanol plant origin.