Integrated multi-omics approach reveals novel associations in the rapeseed diet-microbiota-host axis in pigs.

Diet-mediated host-microbiota interplay is a key factor in optimizing the gut function and overall health of the host. Gaining insight into the biological mechanisms behind this relationship is fundamental to finding sustainable, environment-friendly feed solutions in livestock production systems. Here, we apply a multi-omics integration approach to elucidate sustainable diet-associated host-gut microbiota interactions in pigs and we demonstrate novel and biologically relevant host-microbe associations in the gut, driven by a rapeseed meal-based feed (RSF). Interestingly, RSF-diet promoted the abundance of segmented filamentous bacteria Candidatus Arthromitus that was associated with the maintenance of mucosal immunity in the ileum of pigs. In the colon, RSF diet affected host mRNA splicing functions, which may result in different host gene products, through host-microbiota associations, particularly with the Faecalibacterium population, and through the interaction of dietary components such as sinapic acid with the host cells. Moreover, telomere maintenance and organization functions that may determine the overall health of the host were upregulated and notably associated with Subdoligranulum population in the colon of RSF diet-fed pigs. This integrative multi-omics approach provides more insight into the diet-microbiota-host axis, and a better understanding of mechanisms and opportunities to find new strategies for modulating host health and potentially improving caloric and nutritional efficiency in animal production.

Differential expression of immune-related biomarkers in primary cultures from Atlantic salmon (Salmo salar) exposed to processed Paecilomyces variotii with or without inactivated Moritella viscosa.

Paecilomyces variotii (a filamentous fungus), is a promising novel protein source in fish feeds due to its high nutritional value. Also, P. variotii has Microbial-Associated Molecular Patterns (MAMPs) such as glucans and nucleic acids that could modulate the host's immune response. To understand the potential bioactive properties of this fungus in Atlantic salmon (Salmo salar), our study was conducted to evaluate the gene expression of immune-related biomarkers (e.g., cytokines, effector molecules and receptors) on primary cultures from salmon head kidney (HKLs) and spleen leukocytes (SLs) exposed to either UV inactivated or fractions from P. variotii with or without inactivated Moritella viscosa (a skin pathogen in salmonids). Moreover, the effect of the fermentation conditions and down-stream processing on the physical ultrastructure and cell wall glucan content of P. variotii was characterized. The results showed that drying had a significant effect on the cell wall ultrastructure of the fungi and the choice of fermentation has a significant effect on the quantity of ß-glucans in P. variotii. Furthermore, stimulating Atlantic salmon HKLs and SLs with P. variotii and its fractions induced gene expression related to pro-inflammatory (tnfα, il1ß) and antimicrobial response (cath2) in HKLs, while response in SLs was related to both pro-inflammatory and regulatory response (tnfα, il6 and il10). Similarly, the stimulation with inactivated M. viscosa alone led to an up-regulation of genes related to pro-inflammatory (tnfα, il1ß, il6) antimicrobial response (cath2), intra-cellular signalling and recognition of M. viscosa (sclra, sclrb) and a suppression of regulatory response (il10) in both HKLs and SLs. Interestingly, the co-stimulation of cells with P. variotii and M. viscosa induced immune homeostasis (il6, tgfß) and antimicrobial response (cath2) in SLs at 48h. Thus, P. variotii induces immune activation and cellular communication in Atlantic salmon HKLs and SLs and modulates M. viscosa induced pro-inflammatory responses in SLs. Taken together, the results from physical and chemical characterization of the fungi, along with the differential gene expression of key immune biomarkers, provides a theoretical basis for designing feeding trials and optimize diets with P. variotii as a functional novel feed ingredient for Atlantic salmon.

Cyberlindnera jadinii yeast as a functional protein source: Modulation of immunoregulatory pathways in the intestinal proteome of zebrafish (Danio rerio).

Yeasts contain bioactive components that can enhance fish immune robustness and disease resistance. Our study focused on analyzing intestinal immunoregulatory pathways in zebrafish (Danio rerio) using iTRAQ and 2D LC-MS/MS to quantify intestinal proteins. Zebrafish were fed either control diet (C) or diet supplemented with autolyzed Cyberlindnera jadinii (ACJ). KEGG analysis revealed that ACJ yeast diet induced increased abundance of proteins related to arginine and proline metabolism, phagosome, C-lectin receptor signaling, ribosome and PPAR signaling pathways, which can modulate and enhance innate immune responses. ACJ yeast diet also showed decreased abundance of proteins associated with inflammatory pathways, including apoptosis, necroptosis and ferroptosis. These findings indicate boosted innate immune response and control of inflammation-related pathways in zebrafish intestine. Our findings in the well annotated proteome of zebrafish enabled a detailed investigation of intestinal responses and provide insight into health-beneficial effects of yeast species C. jadinii, which is relevant for aquaculture species.

Effect of Cyberlindnera jadinii yeast on growth performance, nutrient digestibility, and gut health of broiler chickens from 1 to 34 d of age.

The effect of dietary graded levels of Cyberlindnera jadinii yeast (C. jadinii) on growth performance, nutrient digestibility, and gut health of broilers was evaluated from 1 to 34 d of age. A total of 360 male broiler chicks were randomly allocated to 1 of 4 dietary treatments (6 replicate pens each) consisting of a wheat-soybean meal-based pelleted diet (Control or CJ0), and 3 diets in which 10% (CJ10), 20% (CJ20), and 30% (CJ30) of the crude protein were supplied by C. jadinii, by gradually replacing protein-rich ingredients. Body weight and feed intake were measured at d 1, 11, 22, and 32. Pellet temperature, durability, and hardness increased linearly (P < 0.05) with C. jadinii inclusion, with highest (P < 0.05) values for CJ30. Up until d 22, feed conversion ratio (FCR) was similar between treatments (P = 0.169). Overall, increasing C. jadinii inclusion linearly increased (P = 0.047) feed intake but had no effect on weight gain or mortality. FCR increased (P < 0.05) linearly with increasing C. jadinii inclusion but only birds fed CJ30 had a significantly poorer FCR compared to the Control. Ileal digestibility was not affected by C. jadinii inclusion, however, there was a significant linear decrease in crude protein and phosphorus, and a tendency for a decrease in fat digestibility. Apparent metabolizable energy (AME) decreased (P < 0.001) quadratically with increasing C. jadinii and was significantly lower in CJ30 compared to the Control. Ileal concentrations of volatile fatty acids (VFAs) were not affected by C. jadinii inclusion, but butyric acid and total VFAs were linearly and quadratically increased and were significantly higher in cecal digesta of birds fed CJ20 and CJ30. Increasing C. jadinii inclusion was associated with an increase (P < 0.05) in the relative abundance of lactobacillus in the ileum and cecum. In conclusion, C. jadinii yeast can supply up to 20% of the total dietary protein without negatively affecting performance, digestibility, or gut health of broilers. The potential confounding role of feed processing and C. jadinii cell wall components on broiler performance is discussed.

Sugar Kelp (Saccharina latissima) Seaweed Added to a Growing-Finishing Lamb Diet Has a Positive Effect on Quality Traits and on Mineral Content of Meat.

Supplementing ruminants' diet with seaweed has shown positive effect on meat quality and micronutrients important for human health. The objective of the present study was to investigate the use of Saccharina latissima in a lamb diet to improve the eating quality and nutritional value of meat. Six-month-old female Norwegian White lambs (n = 24) were fed, 35 days pre-slaughter, three different diets: a control (CON) and two seaweed diets (SW); supplemented with either 2.5% (SW1) or 5% (SW2). The quality properties of longissimus thoracis et lumborum (LTL) and semimembranosus with adductor (SM+ADD) muscles were examined. The dietary inclusion of seaweed reduced cooking loss and shear force of lamb meat, although the effect was not significant at both supplementation levels. SW1 fed lambs showed a significantly (p < 0.05) improved meat color stability and antioxidant potential. Seaweed also reduced lipid oxidation (TBARS) and the warm-over flavor in SM+ADD compared to the CON lamb. Seaweed fed lambs showed an increased content of selenium and iodine in LTL, thereby fulfilling the requirements for the label "source of nutrient" and "significant source of nutrient", respectively. An increased arsenic content in LTL was, however, also observed with seaweed inclusion (to 1.54 and 3.09 µg/100 g in SW1 and SW2 group, respectively). While relevant positive effects were found in meat using seaweed in lamb feed, some optimization of this feed approach will be desirable.

Cyberlindnera jadinii yeast as a functional protein source for Atlantic salmon (Salmo salar L.): Early response of intestinal mucosal compartments in the distal intestine.

The interplay between nutrition and the immune system is well recognized, and several studies show that experimental diets elicit local morphological changes and alteration of gene and protein expression in the intestinal mucosa of Atlantic salmon. In this study the pathophysiological effects of experimental diets on mucosal responses in the distal intestine of Atlantic salmon were investigated. Atlantic salmon were fed diets with inclusion of soybean meal (SBM) and Cyberlindnera jadinii (CJ) yeast for 7 days. A standard fish meal (FM) diet was used as a control. Morphological, immunohistochemical and gene expression analyses were used to evaluate the presence of immune cells, proliferating cells, and stem cell populations in mucosal compartments of the simple folds in the distal intestine. Fish fed SBM developed morphological changes consistent with SBM induced enteritis. Immunohistochemistry showed an increased presence of apoptotic cells, CD3ϵ and CD8α labelled cells in the simple fold epithelium of SBM group compared with the CJ group. For the investigated genes, expression levels in all three groups were mostly higher in the epithelial compartment of the simple fold than in the compartment beneath the folds. Most changes within the epithelial compartment were observed in fish fed SBM, where expression of CD3ζ, CD8α, MHC I and MHC II were lower than the FM control group. The CJ group had an increased expression of the stem cell marker Lgr5 in the epithelial compartment compared with SBM group. The division of the simple fold into an apical and basal compartment showed that the increase in Lgr5 was evident along the whole length of the simple folds and not confined to the base of the folds. Similarly, proliferation (PCNA, MCM2) and apoptosis (Caspase-3) gene expression was present in the entire length of the simple folds, suggesting that intestinal epithelial cell turnover is not confined to the basal or apical part of the fold. This study shows that the epithelial compartment is active in the early immunoregulatory response towards dietary stimuli and that the level of an intestinal stem cell marker in salmon was influenced by a diet containing CJ yeast.

From a cell model to a fish trial: Immunomodulatory effects of heat-killed Lactiplantibacillus plantarum as a functional ingredient in aquafeeds for salmonids.

Paraprobiotics (dead/inactivated probiotics) are promising candidates in functional feeds to promote growth performance, modulate intestinal microbiota and enhance immune response of fish. During industrial production, fish are exposed to several stressful conditions such as handling, sub-optimal nutrition and diseases that can lead to reduced growth, increased mortalities and large economical losses. Such problems can be mitigated by use of functional feeds, leading to more-sustainable aquaculture and improved animal welfare. Lactiplantibacillus plantarum strain L-137 is a common bacterium found in fermented Southeast Asian dish made from fish and rice. The benefits of its heat-killed form (HK L-137) related to growth performance and immunomodulation have been studied in farmed fish such as Nile Tilapia (Oreochromis niloticus), striped catfish (Pangasianodon hypophthalmus) and bighead catfish (Clarias macrocephalus). To study if such benefits can also be observed in salmonids, we worked both at in vitro level using an intestinal epithelium cell line from rainbow trout (Oncorhynchus mykiss; RTgutGC) stimulated with HK L-137 (Feed LP20™) and at in vivo level with pre-smolt Atlantic salmon (Salmo salar) fed HK L-137 at different inclusion levels (20, 100 and 500 mg of Feed LP20™ kg-1 feed). In RTgutGC, the results showed that the barrier function of the cell monolayer was strengthened along with an increased production of IL-1ß and a decreased production of Anxa1, indicating a modulation of the immune response. Interestingly, a similar trend was detected at the in vivo level in distal intestine from fish fed the highest inclusion level of HK L-137. Here, a lower production of Anxa1 was also detected (after a 61-day feeding period) in addition to an increase of total plasma IgM in the same group. Furthermore, the RNA-seq analysis showed that HK L-137 was able to modulate the gene expression of pathways related to molecular function, biological process and cellular component in distal intestine, without compromising fish performance and gut microbiota. Taken together, our study has shown that HK L-137 can modulate physiological response of Atlantic salmon, making fish more robust against stressful conditions during production.

SALARECON connects the Atlantic salmon genome to growth and feed efficiency.

Atlantic salmon (Salmo salar) is the most valuable farmed fish globally and there is much interest in optimizing its genetics and rearing conditions for growth and feed efficiency. Marine feed ingredients must be replaced to meet global demand, with challenges for fish health and sustainability. Metabolic models can address this by connecting genomes to metabolism, which converts nutrients in the feed to energy and biomass, but such models are currently not available for major aquaculture species such as salmon. We present SALARECON, a model focusing on energy, amino acid, and nucleotide metabolism that links the Atlantic salmon genome to metabolic fluxes and growth. It performs well in standardized tests and captures expected metabolic (in)capabilities. We show that it can explain observed hypoxic growth in terms of metabolic fluxes and apply it to aquaculture by simulating growth with commercial feed ingredients. Predicted limiting amino acids and feed efficiencies agree with data, and the model suggests that marine feed efficiency can be achieved by supplementing a few amino acids to plant- and insect-based feeds. SALARECON is a high-quality model that makes it possible to simulate Atlantic salmon metabolism and growth. It can be used to explain Atlantic salmon physiology and address key challenges in aquaculture such as development of sustainable feeds.

Dietary Inclusion of Hydrolyzed Debaryomyces hansenii Yeasts Modulates Physiological Responses in Plasma and Immune Organs of Atlantic Salmon (Salmo salar) Parr Exposed to Acute Hypoxia Stress.

Stress related to salmon aquaculture practices (handling, sub-optimal nutrition, diseases, and environmental problems) may compromise fish welfare. This study describes the effects of two hydrolyzed Debaryomyces hansenii yeast-based products (LAN4 and LAN6) on physiological and immune responses of Atlantic salmon (Salmo salar) parr exposed to short hypoxia stress. A commercial-like diet (control diet: CD) and two experimental diets (CD supplemented with 0.1% of either component LAN4 or LAN6) were fed to fish for 8 weeks. At the end of the feeding experiment, fish were exposed to 1-min hypoxia and samples were collected at 0, 1, 3, 6, 12, and 24 h post-stress. Results showed that plasma cortisol reached a peak at 1 h post-stress in CD and LAN6 groups, whereas no significant increase in cortisol levels was detected in the LAN4 group. Moreover, the LAN6 group enhanced IL-10 responses to hypoxia, when compared to the control and LAN4 group. This suggests a regulation of immunosuppressive profiles in fish fed LAN4. Hypoxia stress increased TNFα in all groups, which indicates that fish may compensate for the short-term stress response, by modulating innate immune molecules. The apparent suppression of hypoxia responses in the LAN4 group coincided with the detection of differences in goblet cells size and Muc-like proteins production in DI; and upregulation (1 h post-stress) of pathways related to oxygen transport, hemoglobin complex, and glutathione transferase activity and the downregulation of fatty acid metabolism (6 h post-stress) in gills. To conclude, a 1-min hypoxia stress exposure affects the response to stress and immunity; and D. hansenii-based yeast products are promising components in functional aquafeeds for salmon due to their ability to counteract possible consequences of hypoxic stress.

Understanding metabolic phenomena accompanying high levels of yeast in broiler chicken diets and resulting carcass weight and meat quality changes.

The use of yeast as a protein source was investigated in broiler chicken diets on carcass quality, storage stability, and metabolite changes in leg meat. Male Ross 308 chickens (n = 100) were fed with one of 5 diets: control, control added 0.6% formic acid, or 3 diets where soybean meal was substituted with 10, 20, and 30% crude protein from inactivated yeast Cyberlindnera jadinii (CJ10, CJ20, CJ30, respectively). The yeast-containing diets reduced carcass weight, linoleic acid, and warm-over flavor in chicken leg meat. Protein degradation-related metabolite biomarkers were upregulated in the leg of chickens that were fed yeast-containing diets, indicating an adaptive response to the loss of appetite. Chill-stored leg meat of birds fed yeast diets showed increased browning and metallic taste compared with those fed the control diet. The use of formic acid in the diet reduced cooking loss and had a positive effect on vitamin B content.

The Spleen as a Target to Characterize Immunomodulatory Effects of Down-Stream Processed Cyberlindnera jadinii Yeasts in Atlantic Salmon Exposed to a Dietary Soybean Meal Challenge.

Aquaculture feeds have changed dramatically from being largely based on fishmeal (FM) towards increased use of plant protein sources, which could impact the fish's immune response. In order to characterize immunomodulatory properties of novel functional ingredients, this study used four diets, one based on FM, a challenging diet with 40% soybean meal (SBM), and two diets containing 40% SBM with 5% of Cyberlindnera jadinii yeast exposed to different down-stream processing conditions: heat-inactivated (ICJ) or autolysation (ACJ). The immunomodulatory effects of the diets were analyzed in the spleen of Atlantic salmon after 37 days of feeding, using a transcriptomic evaluation by RNA sequencing (RNA-seq) and the detection of specific immunological markers at the protein level through indirect Enzyme-linked Immunosorbent Assay (indirect ELISA). The results showed that SBM (compared to FM) induced a down-regulation of pathways related to ion binding and transport, along with an increase at the protein level of pro-inflammatory cytokines such as tumor necrosis factor alpha (TNFα) and interferon gamma (IFNγ). On the other hand, while ICJ (compared to FM-group) maintain the inflammatory response associated with SBM, with higher levels of TNFα and IFNγ, and with an upregulation of creatine kinase activity and phosphagen metabolic process, the inclusion of ACJ was able to modulate the response of Atlantic salmon compared to fish fed the SBM-diet by the activation of biological pathways related to endocytosis, Pattern recognition receptor (PPRs)-signal transduction and transporter activity. In addition, ACJ was also able to control the pro-inflammatory profile of SBM, increasing Interleukin 10 (IL-10) levels and decreasing TNFα production, triggering an immune response similar to that of fish fed an FM-based diet. Finally, we suggest that the spleen is a good candidate to characterize the immunomodulatory effects of functional ingredients in Atlantic salmon. Moreover, the inclusion of ACJ in fish diets, with the ability to control inflammatory processes, could be considered in the formulation of sustainable salmon feed.

Fecal Excretion and Whole-Body Retention of Macro and Micro Minerals in Atlantic Salmon Fed Torula Yeast Grown on Sugar Kelp Hydrolysate.

Yeast is a microbial feed ingredient that can be produced from non-food biomasses. Brown seaweed contains high levels of complex carbohydrates that are not digested to any extent by monogastric animals but can be used as carbon sources for yeast production. The objective of this study was to investigate how minerals originating from brown macroalgae (Saccharina latissima) are incorporated in Cyberlindnera jadinii yeast and to assess the bioavailability of these different minerals as well as their accumulation into different organs of Atlantic salmon. The yeast C. jadinii was produced on a seaweed hydrolysate mixed with a sugar-rich wood hydrolysate in a 9:1 volume ratio and fed to Atlantic salmon (Salmo salar) in two different experiments: a digestibility experiment with 30% dietary inclusion of yeast and a retention experiment with increasing inclusion of yeast (5, 10, and 20%). Seaweed minerals such as zinc (Zn), copper (Cu), iodine (I), manganese (Mn), and cobalt (Co) were incorporated to a high degree in the yeast. The apparent fecal excretion of minerals was similar in both experiments, in general, with low excretion of, I, bromine (Br), and arsenic (As) (ranging from 18.0% to 63.5%) and high excretion of iron (Fe), Cu, Mn, aluminum (Al), cadmium (Cd) and lead (Pb) (ranging from 56.9% to <100%), despite the different fish size and fecal sampling method. High levels of Cu, I, Br, and Co in the yeast resulted in a linear decrease (p < 0.05) in retention of these minerals in salmon fed increasing levels of yeast. Despite increasing amounts of these minerals in the feed, whole-body levels of Cu and Mn remained stable, whereas whole-body levels of Co, somewhat unexpectedly, decreased with increased dietary yeast inclusion. The Cd from the yeast had low bioavailability but was concentrated more in the kidney (0.038 mg kg-1) and liver (0.025 mg kg-1) than in muscle (0.0009 mg kg-1). The given Cd level in fish strengthens the indication that it is safe to feed salmon with up to 20% inclusion of seaweed yeast without exceeding the maximum limit for Cd of 0.05 mg kg-1 w.w. in fish meat. The level and retention (p < 0.05) of As were lower in the yeast compared to fishmeal. The high level of iodine in S. latissima (3900 mg kg-1) was partly transferred to the yeast, and salmon fed increasing levels of yeast displayed a linear increase in whole-body I content (p < 0.05). There is, however, a need for a growth experiment with larger fish to draw any firm conclusions regarding food safety. Overall, this study shows that yeast grown on hydrolyzed seaweed can be a suitable mineral source for Atlantic salmon, especially when diets are low in fishmeal.

Impact of down-stream processing on functional properties of yeasts and the implications on gut health of Atlantic salmon (Salmo salar).

Yeasts are becoming popular as novel ingredients in fish feeds because of their potential to support better growth and concomitantly ensure good fish health. Here, three species of yeasts (Cyberlindnera jadinii, Blastobotrys adeninivorans and Wickerhamomyces anomalus), grown on wood sugars and hydrolysates of chicken were subjected to two down-stream processes, either direct heat-inactivation or autolysis, and the feed potential of the resulting yeast preparations was assessed through a feeding trial with Atlantic salmon fry. Histological examination of distal intestine based on widening of lamina propria, showed that autolyzed W. anomalus was effective in alleviating mild intestinal enteritis, while only limited effects were observed for other yeasts. Our results showed that the functionality of yeast in counteracting intestinal enteritis in Atlantic salmon was dependent on both the type of yeast and the down-stream processing method, and demonstrated that C. jadinii and W. anomalus have promising effects on gut health of Atlantic salmon.

Rapeseed-based diet modulates the imputed functions of gut microbiome in growing-finishing pigs.

Rapeseed meal is a sustainable feed ingredient that can be used as an alternative to imported soybean meal in European pig production. The gut microbiota plays an important role on pig physiology and health but the impact on microbiota of using rapeseed in diets is still not well known. In this study, 84 purebred Norwegian Landrace pigs with average initial weight of 25 kg were divided into two groups and fed for approximately three months with either a control diet containing soybean meal (CON) or a high-fiber experimental diet where 20% rapeseed meal (RSF) was included as an alternative to soybean meal in CON. The composition and function of microbiome in gut digesta samples were analyzed by performing 16S rRNA gene sequencing and culturing of bacteria. The microbiota diversity and composition were similar between the dietary treatments; however, relative abundance of a variety of bacterial groups and imputed functions of microbiome in the ileum and large intestine were altered when the pigs were fed with a rapeseed-based diet. It was notable that the immune-inducing bacterial group Mucispirillum and anti-inflammatory stimulating bacteria Lachnospira were more abundant in the ileum and large intestine of the RSF group, respectively. Moreover, there was a higher abundance of major amino acid fermenters and amylolytic bacteria in the CON group and a high abundance of putative short chain fatty acid producers in RSF group. In comparison with the CON group, the gut microbiome of RSF group possessed an enhanced potential for carbohydrate and energy metabolism and a reduced potential for bacterial pathogenicity-related pathways.

Replacing soybean meal with rapeseed meal and faba beans in a growing-finishing pig diet: Effect on growth performance, meat quality and metabolite changes.

Rapeseed meal and faba beans (RSM/FB) can serve as an alternative to imported soybean meal (SBM). In this study, forty Norwegian crossbred ([Landrace x Yorkshire] x Duroc) growing-finishing pigs (108.7 ±â€¯4.2 kg final BW) were fed a diet with either SBM or RSM/FB as protein sources. RSM/FB increased feed conversion ratio (P = .04) in the finishing period, reduced lightness (P = .04) and yellowness (P = .004) of meat, changed amounts of individual fatty acids, but not of total SFA, MUFA and PUFA. Importantly, RSM/FB reduced the glucose level (P < .05) in meat. Lower pyroglutamic acid (P = .06) in RSM/FB indicate lower oxidative stress in pre-rigor muscle cell. Increased abundance of free amino acids, sweet tasting metabolites, reduced warmed-over flavor and flavor attributes indicated desirable properties of RSM/FB meat. To conclude, RSM/FB in pig diet supported growth performance and carcass quality comparable to SBM and had a positive effect on meat quality.

Candida utilis yeast as a functional protein source for Atlantic salmon (Salmo salar L.): Local intestinal tissue and plasma proteome responses.

Microbial ingredients such as Candida utilis yeast are known to be functional protein sources with immunomodulating effects whereas soybean meal causes soybean meal-induced enteritis in the distal intestine of Atlantic salmon (Salmo salar L.). Inflammatory or immunomodulatory stimuli at the local level in the intestine may alter the plasma proteome profile of Atlantic salmon. These deviations can be helpful indicators for fish health and, therefore potential tools in the diagnosis of fish diseases. The present work aimed to identify local intestinal tissue responses and changes in plasma protein profiles of Atlantic salmon fed inactive dry Candida utilis yeast biomass, soybean meal, or combination of soybean meal based diet with various inclusion levels of Candida utilis. A fishmeal based diet was used as control diet. Inclusion of Candida utilis yeast to a fishmeal based diet did not alter the morphology, immune cell population or gene expression of the distal intestine. Lower levels of Candida utilis combined with soybean meal modulated immune cell populations in the distal intestine and reduced the severity of soybean meal-induced enteritis, while higher inclusion levels of Candida utilis were less effective. Changes in the plasma proteomic profile revealed differences between the diets but did not indicate any specific proteins that could be a marker for health or disease. The results suggest that Candida utilis does not alter intestinal morphology or induce major changes in plasma proteome, and thus could be a high-quality alternative protein source with potential functional properties in diets for Atlantic salmon.

Effects of long-term feeding of rapeseed meal on skeletal muscle transcriptome, production efficiency and meat quality traits in Norwegian Landrace growing-finishing pigs.

This study was performed to investigate the effects of dietary inclusion of 20% rapeseed meal (RSM) as an alternative to soybean meal (SBM) in a three-month feeding experiment with growing finishing pigs. Dietary alteration affected growth performance, several carcass traits and transcriptional responses in the skeletal muscle, but did not affect measured meat quality traits. In general, pigs fed the RSM test diet exhibited reduced growth performance compared to pigs on SBM control diet. Significant transcriptional changes in the skeletal muscle of growing pigs fed RSM diet were likely the consequence of an increased amount of fiber and higher polyunsaturated fatty acids, and presence of bioactive phytochemicals, such as glucosinolates. RNAseq pipeline using Tophat2-Cuffdiff identified 57 upregulated and 63 downregulated genes in RSM compared to SBM pigs. Significantly enriched among downregulated pathways was p53-mediated signalling involved in cellular proliferation, while activation of negative growth regulators (IER5, KLF10, BTG2, KLF11, RETREG1, PRUNE2) in RSM fed pigs provided further evidence for reduced proliferation and increased cellular death, in accordance with the observed reduction in performance traits. Upregulation of well-known metabolic controllers (PDK4, UCP3, ESRRG and ESRRB), involved in energy homeostasis (glucose and lipid metabolism, and mitochondrial function), suggested less available energy and nutrients in RSM pigs. Furthermore, several genes supported more pronounced proteolysis (ABTB1, OTUD1, PADI2, SPP1) and reduced protein synthesis (THBS1, HSF4, AP1S2) in RSM muscle tissue. In parallel, higher levels of NR4A3, PDK4 and FGF21, and a drop in adropin, ELOVL6 and CIDEC/FSP27 indicated increased lipolysis and fatty acid oxidation, reflective of lower dressing percentage. Finally, pigs exposed to RSM showed greater expression level of genes responsive to oxidative stress, indicated by upregulation of GPX1, GPX2, and TXNIP.

Identification of Sinapine-Derived Choline from a Rapeseed Diet as a Source of Serum Trimethylamine N-Oxide in Pigs.

Choline and its metabolites have diverse and important functions in many physiological processes, especially for anabolic metabolism in growth and reproduction. Besides endogenous biosynthesis and direct choline supplement, choline esters in the diet are another source of choline in the body. Phenolic choline esters are a group of unique dietary choline esters rich in the seeds of Brassicaceae plants, among which sinapine is a choline ester of sinapic acid abundant in rapeseed. In this study, 40 nursery pigs were fed with rapeseed-derived feed ingredients (RSF) or soybean meal for 3 weeks (20 pigs/diet). The metabolic fate of sinapine-derived choline in RSF was examined by comparing the distribution of choline and its metabolites in digesta, liver, and serum samples by liquid chromatography-mass spectrometry analysis. The results showed that choline was released from extensive hydrolysis of sinapine in the small intestine. However, sinapine-derived choline did not increase the levels of choline and its major metabolites, including betaine, phosphocholine, and glycerophosphocholine, in the liver and serum. Instead, RSF feeding increased trimethylamine (TMA), the microbial metabolite of choline, in the large intestine and further increased trimethylamine N-oxide (TMAO), the oxidation metabolite of TMA, in the liver and serum. Overall, these results suggested that sinapine-derived choline from rapeseed feeding had limited influences on the post-absorption choline pool as a result of its low bioavailability but may serve as a major source of TMAO through microbial metabolism in nursery pigs. Improving the bioavailability of sinapine-derived choline might have the potential to modify the nutritional values and functionalities of rapeseed meal in swine feeding.

Selection for feed efficiency in Atlantic salmon using individual indicator traits based on stable isotope profiling.

BACKGROUND: We used stable isotope profiling (15N and 13C) to obtain indicator phenotypes for feed efficiency in aquaculture. Our objectives were to (1) examine whether atom percent of stable isotopes of nitrogen and carbon can explain more of the variation in feed conversion ratio than growth alone, and (2) estimate the heritabilities of and genetic correlations between feed efficiency, growth and indicator traits as functions of nitrogen and carbon metabolism in various tissues. A 12-day experiment was conducted with 2281 Atlantic salmon parr, with an average initial weight of 21.8 g, from 23 full-sib families that were allocated to 46 family tanks and fed an experimental diet enriched with 15N and 13C. RESULTS: Using leave-one-out cross-validation, as much as 79% of the between-tank variation in feed conversion ratio was explained by growth, indicator traits, and sampling day, compared to 62% that was explained by growth and sampling day alone. The ratio of tissue metabolism, estimated by a change in isotope fractions relative to body growth, was used as an individual indicator for feed efficiency. For these indicator ratio traits, the estimated genetic correlation to feed conversion ratio approached unity but their heritabilities were low (0.06 to 0.11). These results indicate that feed-efficient fish are characterized by allocating a high fraction of their metabolism to growth. Among the isotope indicator traits, carbon metabolism in the liver had the closest estimated genetic correlation with feed conversion ratio on a tank level (- 0.9) but a low estimated genetic correlation with individually recorded feed efficiency indicator ratio traits. The underlying determinants of these correlations are largely unknown. CONCLUSIONS: Our findings show that the use of indicator ratio traits to assess individual feed efficiency in Atlantic salmon has great prospects in selection programs. Given that large quantities of feeds with contrasting isotope profiles of carbon and/or nitrogen can be produced cost-effectively, the use of stable isotopes to monitor nitrogen and carbon metabolism in various tissues has potential for large-scale recording of individual feed efficiency traits, without requiring individual feed intake to be recorded.