Impact of Dietary Supplementation with Sodium Butyrate Protected by Medium-Chain Fatty Acid Salts on Gut Health of Broiler Chickens

This research was co-funded by the European Fund of Regional Development of the European Union within the framework of the FEDER operating program of Catalunya 2014-2020 (project COMRDI16-1-0033) and managed by ACCIÓ.Nutritional strategies to improve gut health of broilers are under research. This study investigated the effect of dietary supplementation with sodium butyrate protected by sodium salts of medium-chain fatty acids as a feed additive on broiler gut health. The first experiment was conducted to evaluate the effect of supplementing at 0.5, 1, and 2 kg/t in broilers housed under optimal conditions. Supplementation at 0.5 and 1 kg/t maintained goblet cell counts at 10 days of age (p ≤ 0.05), and supplementation at 1 kg/t decreased intraepithelial lymphocyte counts compared to 2 kg/t at 39 days (p ≤ 0.10). Abdominal fat pad levels of lauric and myristic acids were gradually increased by supplement dose (p ≤ 0.05). In the second experiment, the feed additive at 1 kg/t was evaluated in coccidiosis-challenged broilers. Experimental treatments were as follows: non-challenged, control-challenged, and supplemented-challenged treatments. Coccidiosis negatively impact performance and modify histomorphometry and microbiota (p ≤ 0.05). The feed additive increased crypt depth at 7 days post-inoculation and goblet cell count at 14 days post-inoculation (p ≤ 0.05). Further, supplementation interacted with the microbiota modification led by the coccidiosis (p ≤ 0.05). These results suggest that this feed additive could be a useful strategy to reinforce the gut barrier, especially for birds under coccidiosis-challenge treatments

Under control: how a dietary additive can restore the gut microbiome and proteomic profile, and improve disease resilience in a marine teleostean fish fed vegetable diets

[Background]: The constant increase of aquaculture production and wealthy seafood consumption has forced the industry to explore alternative and more sustainable raw aquafeed materials, and plant ingredients have been used to replace marine feedstuffs in many farmed fish. The objective of the present study was to assess whether plant-based diets can induce changes in the intestinal mucus proteome, gut autochthonous microbiota and disease susceptibility of fish, and whether these changes could be reversed by the addition of sodium butyrate to the diets. Three different trials were performed using the teleostean gilthead sea bream (Sparus aurata) as model. In a first preliminary short-term trial, fish were fed with the additive (0.8%) supplementing a basal diet with low vegetable inclusion (D1) and then challenged with a bacteria to detect possible effects on survival. In a second trial, fish were fed with diets with greater vegetable inclusion levels (D2, D3) and the long-term effect of sodium butyrate at a lower dose (0.4%) added to D3 (D4 diet) was tested on the intestinal proteome and microbiome. In a third trial, the long-term effectiveness of sodium butyrate (D4) to prevent disease outcome after an intestinal parasite (Enteromyxum leei) challenge was tested. [Results]: The results showed that opposed forces were driven by dietary plant ingredients and sodium butyrate supplementation in fish diet. On the one hand, vegetable diets induced high parasite infection levels that provoked drops in growth performance, decreased intestinal microbiota diversity, induced the dominance of the Photobacterium genus, as well as altered the gut mucosal proteome suggesting detrimental effects on intestinal function. On the other hand, butyrate addition slightly decreased cumulative mortality after bacterial challenge, avoided growth retardation in parasitized fish, increased intestinal microbiota diversity with a higher representation of butyrate-producing bacteria and reversed most vegetable diet-induced changes in the gut proteome. [Conclusions]: This integrative work gives insights on the pleiotropic effects of a dietary additive on the restoration of intestinal homeostasis and disease resilience, using a multifaceted approach.This work has been carried out with financial support from the European Union under grant projects ARRAINA (FP7-KBBE-2011-288,925) to JPS, MP and VK and ParaFishControl (H2020-634429) to ASB. Additional funding has been received from Spanish Ministry of Economy and Competitiveness (MINECO) project no. AGL2013- 48560-R to JPS and ASB, and Generalitat Valenciana (PROMETEOII/2014/085) to ASB. MCP was contracted under CSIC PIE project no. 201740E013 and MINECO FPDI-2013-15741, and IE under APOSTD/2016/037 grant by the “Generalitat Valenciana”.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)

Short Communication : Evaluation of Intestinal Release of Butyric Acid from Sodium Butyrate Protected by Salts of Medium-Chain Fatty Acids in Broiler Chickens

Butyric acid has received great attention as a feed additive to maintain or increase the gut integrity and health of broiler chickens. Particularly, the protection of butyrate is under research to allow slow intestinal release of butyric acid and to promote its beneficial effects throughout the intestine. This study evaluated in vivo the intestinal release of butyric acid from sodium butyrate protected by salts of medium-chain fatty acid in broilers. Brilliant blue was used as an inert marker, so it was included in the feed additive that broilers ingested for two days. The gastrointestinal tract was then colored in blue from jejunum and backward. Considering the digesta color of the broilers non-supplemented as blank, it allowed quantification of the amount of brilliant blue, and consequently, butyric acid delivered in the intestine from the protected feed additive. Few traces of butyric acid were released in the duodenum and proximal jejunum, whereas the major amount (45.9%) was delivered in the distal ileum (p < 0.001). These results suggest that this in vivo approach allows for evaluation of the intestinal delivery of butyric acid supplemented as protected sodium butyrate by medium-chain fatty acids, showing a gradual intestinal release of butyric acid in broiler chickens

Short Communication: Evaluation of Intestinal Release of Butyric Acid from Sodium Butyrate Protected by Salts of Medium-Chain Fatty Acids in Broiler Chickens

Butyric acid has received great attention as a feed additive to maintain or increase the gut integrity and health of broiler chickens. Particularly, the protection of butyrate is under research to allow slow intestinal release of butyric acid and to promote its beneficial effects throughout the intestine. This study evaluated in vivo the intestinal release of butyric acid from sodium butyrate protected by salts of medium-chain fatty acid in broilers. Brilliant blue was used as an inert marker, so it was included in the feed additive that broilers ingested for two days. The gastrointestinal tract was then colored in blue from jejunum and backward. Considering the digesta color of the broilers non-supplemented as blank, it allowed quantification of the amount of brilliant blue, and consequently, butyric acid delivered in the intestine from the protected feed additive. Few traces of butyric acid were released in the duodenum and proximal jejunum, whereas the major amount (45.9%) was delivered in the distal ileum (p &lt; 0.001). These results suggest that this in vivo approach allows for evaluation of the intestinal delivery of butyric acid supplemented as protected sodium butyrate by medium-chain fatty acids, showing a gradual intestinal release of butyric acid in broiler chickens

Sodium salt medium-chain fatty acids and Bacillus-based probiotic strategies to improve growth and intestinal health of gilthead sea bream (Sparus aurata)

Background. The increased demand for fish protein has led to the intensification of aquaculture practices which are hampered by nutritional and health factors affecting growth performance. To solve these problems, antibiotics have been used for many years in the prevention, control and treatment against disease as well as growth promoters to improve animal performance. Nowadays, the use of antibiotics in the European Union and other countries has been completely or partially banned as a result of the existence of antibiotic cross-resistance. Therefore, a number of alternatives, including enzymes, prebiotics, probiotics, phytonutrients and organic acids used alone or in combination have been proposed for the improvement of immunological state, growth performance and production in livestock animals. The aim of the present study was to evaluate two commercially available feed additives, one based on medium-chain fatty acids (MCFAs) from coconut oil and another with a Bacillus-based probiotic, in gilthead sea bream (GSB, Sparus aurata), a marine farmed fish of high value in the Mediterranean aquaculture. Methods. The potential benefits of adding two commercial feed additives on fish growth performance and intestinal health were assessed in a 100-days feeding trial. The experimental diets (D2 and D3) were prepared by supplementing a basal diet (D1) with MCFAs in the form of a sodium salt of coconut fatty acid distillate (DICOSAN R ; Norel, Madrid, Spain), rich on C-12, added at 0.3% (D2) or with the probiotic Bacillus amyloliquefaciens CECT 5940, added at 0.1% (D3). The study integrated data on growth performance, blood biochemistry, histology and intestinal gene expression patterns of selected markers of intestinal function and architecture. Results. MCFAs in the form of a coconut oil increased feed intake, growth rates and the surface of nutrient absorption, promoting the anabolic action of the somatotropic axis. The probiotic (D3) induced anti-inflammatory and anti-oxidant effects with changes in circulating cortisol, immunoglobulin M, leukocyte respiratory burst, and mucosal expression levels of cytokines, lymphocyte markers and immunoglobulin T. Discussion. MCFA supplementation showed positive effects on GSB growth and intestinal architecture acting mainly in the anterior intestine, where absorption takes place. The probiotic B. amyloliquefaciens CECT 5940 exhibited key effects in the regulation of the immune status inducing anti-inflammatory and anti-oxidant effects which can be potentially advantageous upon infection or exposure to other stressors. The potential effects of these feed additives in GSB are very promising to improve health and disease resistance in aquaculture.Spanish MINECO: AGL2013-48560-R. .Generalitat Valenciana: PROMETEO-FASE II/2014/085.MINECO: FPDI-2013-15741.CSIC PIE: 201740E013Peer reviewe

Impact of low fish meal and fish oil diets on the performance, sex steroid profile and male-female sex reversal of gilthead sea bream (Sparus aurata) over a three-year production cycle

Juveniles of the protandrous hermaphrodite gilthead sea bream (Sparus aurata) were fed from early life stages to completion of sex maturation with plant-based diets formulated by BioMar. Total fish meal (FM) was included at 25% in the control diet (D1) and at 5% in the other three diets (D2, D3, D4). Added oil was either fish oil (FO) (D1) or a blend of vegetable oils replacing 58% (D2) and 84% (D3, D4) of FO. A commercial butyrate preparation (BP-70® NOREL) was added to the D4 diet at 0.4%. All fish grew fast through a three-year ongrowing cycle with overall specific growth rates of 1.4%, 0.7% and 0.5% for fish harvest at 300 g, 1 kg and 1.5–1.7 kg, respectively. Overall feed efficiency decreased progressively as fish size increased from 0.99 in 300 g fish to 0.8–0.7 in 1–1.7 kg fish. At the last stage, a clear sexual dimorphism was found for body weight and hepatosomatic index when all sampled fish were considered as a whole. A sexual dimorphism was also found for sex steroids with a peak of estradiol in the females sampled in October–December, whereas the peak of 11-ketotestosterone was delayed in males to December. Plasma levels of testosterone were similar in both sexes. The two first components of principal component analysis (PCA) explained >90% of total variance of plasma levels of sex steroids. The displacement along X-axis clearly separated males and females, whereas the movement along Y-axis was related to sampling time. An androgenic effect in the steroid plasma profile of fish fed plant-based diets was also shown, which was especially evident for the low FM/FO diet formulation (D3). This effect was reversed by butyrate and the female/male ratio of D4 fish (age class +3) did not differ from that of control fish, whereas the proportion of females in D2 + D3 fish was higher (P < 0.06) than in control fish (64% vs. 50%). This group also showed increased circulating levels of vitellogenin, non-detectable in males/intersex fish of any experimental group. Taken together, well balanced plant-based diets did not compromise the maximum growth of farmed gilthead sea bream over a 3-year production cycle, and data on key performance indicators with a wide range of FM/FO formulations can be considered as reference values for the species. It is noteworthy that the plant-based diets had an impact on sex reversal, but this effect was mitigated by butyrate supplementation.This work has been carried out with financial support from the European Union (ARRAINA, FP7-KBBE-2011-5-288925, Advanced Research Initiatives for Nutrition and Aquaculture). Additional funding was obtained from the Spanish MINECO (MI2-Fish, AGL2013-48560; Pubertrait, AGL2016-75400) and from Generalitat Valenciana (PROMETEO FASE II-2014/085 and 051).Peer reviewe

Trade-off effects of medium-chain fatty acids in the metabolic capacity and swim performance of gilthead sea bream fed different protein dietary sources

Trabajo presentado en el 11º Congreso de la Asociación Ibérica de Endocrinología Comparada (AIEC), celebrado en Vigo (España), del 13 al 15 de julio de 2017Short- and medium-chain fatty acids (SCFAs and MCFAs) are natural compounds present in both animal and plant tissues that play a key role on energy-dependent mitochondrial processes and intracellular signaling. Since cellular uptake and transport of SCFAs and MCFAs across the cell and mitochondria do not require the action of specific transporters, these compounds are considered ideal nutritive sources for a rapid energy uptake and utilisation. Experimental evidences show that inclusion of the C4 carboxylic acid (butyric acid, SCFA) exerts benefits upon the intestine of gilthead sea bream reversing some detrimental effects of extreme plant protein-based diets, but there is no information about the effects of C7 carboxylic acid (heptanoic acid, MCFA) in fish. The aim of this study was to explore the potential benefits of a commercial preparation of heptanoate sodium salt in a factorial design (2 x 2) with standard (D1 and D2) or low (D3 and D4) inclusion levels of fish meal (FM) as dietary protein source in an 11-weeks trial. Histopathological scoring of intestine did not reveal detrimental effects of dietary protein or heptanoate supplementation. Results of growth performance indicated a fast (SGR=2.26-2.32) and efficient (FE=0.91-0.94) growth with no changes in plasma Igf-I levels among fish fed the four experimental diets. Heptanoate supplementation (D2 and D4) increased plasma cortisol regardless of dietary protein source, whereas this MCFA increased plasma Gh concentration in D2 and decreased it in D4 fish. In addition, an improved swim performance evidenced by a greater critical swimming speed (Ucrit) was orchestrated i) by heptanoate in FM-based diet (D2) or ii) by the nutritionally-mediated changes through cortisol and Gh in D3 and D4 fish. In this sense, the reduction of Gh values produced by heptanoate in the low FM diet (D4) can be viewed as a protective measure to counteract disproportionate oxidative metabolic rates.Peer reviewe

Trade-off effects of medium-chain fatty acids in the metabolic capacity and swim performance of gilthead sea bream fed different protein dietary sources

Trabajo presentado en el 11º Congreso de la Asociación Ibérica de Endocrinología Comparada (AIEC), celebrado en Vigo (España), del 13 al 15 de julio de 2017The aim of this study was to evaluate the potential benefits of a commercial preparation of the medium-chain fatty acid (MCFA) heptanoate sodium salt (NOREL, HEPTON®) in a factorial design (2 x 2) with standard (20%, D1 and D2) or low (5%, D3 and D4) inclusion levels of fish meal (FM) as dietary protein source in an 11-weeks gilthead sea bream feeding trial. Heptanoate was added at 0.3% in D2 and D4 diets. Results of growth performance from 13-14 g to 81-84 g indicated a fast (SGR=2.2-2.3%) and efficient (FE=0.91-0.94) growth, with no changes in plasma IGF-I levels among fish fed the four experimental diets. Even so, an early impairment of FE, reversed by heptanoate supplementation, was observed in D1 FM-based diet. No signs of histopathological damage were found in any group, although higher hepatic glycogen depots were observed in fish fed both heptanoate-supplemented diets at the end of the trial. Other lasting heptanoate effects included changes on plasma antioxidant capacity, circulating cortisol and growth hormone (GH) levels, as well as in metabolic capabilities assessed by measures of oxygen consumption (MO2, mgO2/kg/h) and critical swimming speed (Ucrit) in swim performance tests. The pattern of respirometry metabolism closely resembled that of early FE, and the predictive value of plasma GH for different critical swimming speeds at fish exhaustion is suggested. Altogether, these results support the potential use of heptanoate to improve the healthy metabolic condition of farmed fish under sub-optimal rearing conditions.This work has been carried out with financial support from NOREL S.A. and BioMAR. Additional funding has been received by Spanish MINECO project no. AGL2013-48560-R and Generalitat Valenciana (PROMETEO-FASE II/2014/085). M.C.P. was supported by MINECO through grant FPDI-2013-15741 and by CSIC PIE project no. 201740E013. J.A.M.-S. is supported by a Postdoctoral Research Fellowship (Juan de la CiervaFormación, Reference FJCI-2014-20161) from MINECO.Peer reviewe

Dietary sodium heptanoate helps to improve feed efficiency, growth hormone status and swimming performance in gilthead sea bream (Sparus aurata)

The potential benefits of a commercial preparation of heptanoate (NOREL, HEPTON®) were evaluated in an 11‐week gilthead sea bream feeding trial (May–August), using a factorial design with four isoproteic and isoenergetic diets. Fish meal (FM) was added at 200 g/kg in D1–D2 diets and at 50 g/kg in D3–D4 diets, which also contained fish peptones and plant proteins as source of proteins. Heptanoate was added at 3 g/kg in D2 and D4 diets. All fish grew from 13–14 g to 81–84 g with an overall feed efficiency (FE) of 0.91–0.94. An early impairment of FE (weeks 1–4) was found with the standard FM‐based diet (D1), but this detrimental condition was reversed by heptanoate, increasing FE from 0.88 in D1 fish to 0.99 in D2 fish. Further improvements were progressively diluted over time, remaining D2 and D3–D4 fed fish almost undistinguishable through all the trial. Heptanoate supplementation produced higher hepatic glycogen depots, but no signs of histopathological damage were found in liver or intestine. Other lasting heptanoate effects included changes in plasma antioxidant capacity, plasma cortisol and growth hormone levels, and measures of respirometry in swimming performance tests. Altogether, it supports the potential use of heptanoate to speed up adaptive and healthy metabolic states of farmed fish to cope with challenging culture conditions.This work has been carried out with the financial support from NOREL S.A. and BioMar. Additional funding has been received by Spanish MINECO project no. AGL2013‐48560‐R and Generalitat Valenciana (PROMETEO‐FASE II/2014/085). MCP was supported by MINECO through grant FPDI‐2013‐15741 and by CSIC PIE project no. 201740E013. JAMS was supported by a postdoctoral research fellowship (Juan de la Cierva‐Formación, Reference FJCI‐2014‐20161) from MINECOPeer reviewe

Additional file 3: Figure S2. of Under control: how a dietary additive can restore the gut microbiome and proteomic profile, and improve disease resilience in a marine teleostean fish fed vegetable diets

Krona visualisation of the relative abundance of intestinal bacterial OTUs identified in fish fed D1. (HTML 238 kb