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EggMeat Symposia 2013 Bergamo 15-19 September 2013 Comparison between the dietary supplementation of sodium selenite and Selenium-yeast on meat Se accumulation in broiler E. SIMON1*, N. BALLET1, M. FRANCESCH2, J. BRUFAU2 1 Lesaffre Feed Additives – 137, rue Gabriel Péri - 59700 MARCQ EN BAROEUL – France 2 IRTA - Monogastric Nutrition - Mas de Bover - El Morell, Km 3.8 - E-43120 CONSTANTÍ – Spain *Corresponding author: [email protected] Selenium (Se) is an essential nutrient for animals as a component of the enzyme glutathione peroxidase, which plays a key role in the protection of cells against oxidative agents. Depending on the chemical form of the element, its absorption and metabolization is not the same, resulting in different bioavailability and accumulation in tissues and muscle. The objective of this study was to compare the bioavailability of sodium selenite and Selenium –yeast (Se-Yeast) sources added in the feed of broilers. A total of 182 chicks of ROSS 380 strain were allocated in 91 cages, for a total of 7 cages per dietary treatment. From 7 to 37 days of age, they were fed with a cornsoybean meal based diet supplemented with 0, 0.15, 0.30, 0.45 or 1 ppm of Se. Three forms of Se were tested: sodium selenite, Se-yeast (Selsaf –CNCM I-3399) and another Se-yeast. At 37 days of age, blood and pectoral muscle samples were collected from each bird. Se accumulation in blood and pectoral muscle increased linearly (P<0.0001) with the increase of Se intake from both inorganic and organic Se sources. However, the slopes of the regression lines were higher (P<0.001) for organic Se sources compared with sodium selenite. At higher dosage, 0.45 and 1 ppm, Se accumulation in muscle with Selsaf was higher compared with the other Se enriched-yeast (p<0.05). This difference of efficiency among Se sources can be related to their different contents in selenomethionine (63% vs 56.7% of Selenomethionine (Se-Meth) in Selsaf and in the other Seyeast respectively), which may be used as a substitute for methionine in the synthesis of all proteins, particularly in pectoral muscles. By consuming 100 g of meat, the intake of Se with the Se yeast source represents 47 % of the recommended human daily allowance compared to 19 % with the sodium selenite source. Keywords: Se-yeast; bioavailability; meat; broiler. Introduction Se is an essential nutrient for animals and humans as a component of the enzyme glutathione peroxidase (Rotruck et al., 1973), which plays a key role in the protection of cells against oxidative agents (Vamecq, 2004). Se can be naturally present in feedstuffs but its level can be very variable depending on the food source and Se content of the substrate on which the food is grown (Whanger et al., 2002). For this purpose, different source of Se can be provided in the feed to meet nutritional requirements of animals in different chemical forms: Se in its inorganic form (Selenite or Selenate) or in its organic form (Se-Yeast). To produce Se-Yeast, yeast, grown on a culture medium supplemented with selenite, has the ability to metabolize this Se to produce mainly Se in the Se-meth form. In general, adequate Se supplementation is considered to be a crucial factor in maintaining the high productive and reproductive characteristics of commercial poultry. Recent reviews describing into further details the mode of action of Se and its role in the antioxidant system has developed a new interest about the use of Selenium in poultry nutrition (Surai et al., 2002). Depending on the source of World’s Poultry Science Journal, Volume 69, Supplement 1 EggMeat Symposia 2013 Bergamo 15-19 September 2013 Se used and the manufacturing process, the bioavailability of Se can be different. In a previous nutritional study, a higher bioavailability of Se-yeast has been shown compared to the selenite (Yoshida et al., 2002). The bioavailability of Se can be assessed by the tissue accumulation approach, supplementing Se at graded levels in the feed and analysing the slope ratio of the responses to graded levels of supplemented Se (Yoshida et al., 1999). The accumulation of Se in the muscle can have two main interests for the Se-enrichment of animal tissues, providing a source of Se for humans and the improvement of meat quality with a reduction of water drip losses (Downs et al., 2000) and a reduction of the lipid oxidation during meat storage (Kim et al, 2010). The objective of this study was to assess the difference of bioavailability between sodium selenite and Se-Yeast sources added in the feed of broilers. Materials and methods Male broiler chickens Ross 380 were used for the trial. During the first seven days, all chickens were fed with the same diet, based on maize and soybean meal without Se supplementation. At day 7, broilers were allocated into 7 groups according to their live weights, after discarding the small ones. The experiment was set up as a randomized complete block design, with seven blocks, thirteen dietary experimental treatments (table 1) and 7 replicates of 2 chickens per treatment. The 7 weight groups were assigned at random to one block location in the battery cage. In this experiment, 182 broilers were used and they were allocated to 91 cages. A single diet was used during the whole experimental period based on corn and soybean meal. The nutritional properties were 3050 kcal/kg and 21 % of crude protein. Depending on the treatment group, this diet was supplemented with no Se, sodium selenite, Selsaf (CNCM I-3399) or another Se-yeast. Four levels of Se were tested for each source: 0.15, 0.30, 0.45 and 1 mg/kg of feed. The Se-meth contents were different between the Se yeast sources: 63% and 56.7% of the total Se content for the Selsaf and the other Se yeast respectively. The arrangement of treatments was: Dietary treatment T-1 T-2 T-3 T-4 T-5 T-6 T-7 T-8 T-9 T-10 T-11 T-12 T-13 Se Source No Se Sodium selenite Sodium selenite Sodium selenite Sodium selenite Selsaf Selsaf Selsaf Selsaf Other Se-Yeast Other Se-Yeast Other Se-Yeast Other Se-Yeast Level of Se added (mg/kg of feed) 0 0.15 0.30 0.45 1.0 0.15 0.30 0.45 1.0 0.15 0.30 0.45 1.0 Table 1: Se source and quantity of Se tested for each treatment group At day 37, blood samples from all broilers were collected by cardiac puncture. Moreover, after blood sampling, animals were slaughtered to collect pectoral muscle. The samples were pooled per cage to obtain a total of 7 samples of blood and 7 samples of meat per treatment. World’s Poultry Science Journal, Volume 69, Supplement 2 EggMeat Symposia 2013 Bergamo 15-19 September 2013 These samples were sent to the laboratory to analyse the total Se content by ICP-MS (Inductively Coupled Plasma Mass Spectrometer) method. Complete data were analysed as a randomised complete block design with a two-way analysis of variance (block and treatment) using the GLM procedure of SAS (SAS Institute, 2008). The experimental unit was the cage and there were a total of 13 dietary treatments replicated 7 times. Data from T-2 to T-13 treatments were analysed in a 3 x 4 factorial arrangement (three Se sources and four levels of Se supplementation).The model included block, the main effects of Se source and dose of Se supplementation and their interaction. Significance was based on a 5% probability level. Moreover, data from Se measurements in blood and pectoral muscle from sodium selenite, SelSaf and other SeYeast were regressed separately on the total Se intake from 7 to 38 days, estimated from the expected level of Se supplementation and the recorded feed intake, including the 0 supplemental Se level Results and discussion Main effects of Se source and Se concentration (p<0.0001), as well as source x level interactions (p<0.0001) were observed for Se accumulation in blood and pectoral muscle (Table 2). For blood Se accumulation, there were no significant differences between Se sources at the lowest concentration of Se supplementation (0.15 mg/kg). At 0.3, 0.45 and 1 mg/kg of Se addition, blood Se accumulation was higher with organic Se sources compared with inorganic Se (P<0.05). In pectoral muscle, Se accumulation was always higher using organic Se sources (P<0.05). Moreover, at 0.45 and 1 mg/kg of Se addition, Se concentration in muscle was higher with SelSaf compared with the other Se-Yeast (P<0.05). Treatment Group Se Source T-1 T-2 T-3 T-4 T-5 T-6 T-7 T-8 T-9 T-10 T-11 T-12 T-13 Standard error Anova Factorial Analysis Se Source Se level Interaction No Se Sodium selenite Sodium selenite Sodium selenite Sodium selenite Selsaf Selsaf Selsaf Selsaf Other Se-Yeast Other Se-Yeast Other Se-Yeast Other Se-Yeast Level of Se added (mg/kg of feed) 0 0.15 0.30 0.45 1.0 0.15 0.30 0.45 1.0 0.15 0.30 0.45 1.0 Se content in Blood (mg/kg DM) Se content in Pectoral Muscle (mg/kg DM) 0.73i 1.20h 1.33gh 1.50fg 2.10cd 1.29h 1.77e 2.21c 3.29a 1.19h 1.59f 2.00d 2.93b 0.063 0.33i 0.43h 0.46h 0.51gh 0.56g 0.93f 1.37e 1.89c 3.59a 0.84f 1.30e 1.73d 3.29b 0.033 p<0.0001 p<0.0001 p<0.0001 p<0.0001 p<0.0001 p<0.0001 p<0.0001 p<0.0001 Table 2 : Effect of Se source and dietary supplement level of Se on blood and pectoral muscle accumulation at 37 days a-h Means within column with different superscript differ significantly (p<0.05) World’s Poultry Science Journal, Volume 69, Supplement 3 EggMeat Symposia 2013 Bergamo 15-19 September 2013 Se accumulation in blood and pectoral muscle increased linearly (P<0.0001) with the increase of Se intake from both inorganic and organic Se sources (Figure 1 A and B). However, the slopes of the regression lines were higher (P<0.001) for SelSaf and Se-Yeast compared with sodium selenite. There were no significant differences in the slopes of regression lines when Se intake from SelSaf or SeYeast was regressed on Se accumulation in blood and pectoral muscle. A/ B/ Figure 1: Effect of Se intake from different sources on blood (A) and pectoral muscle (B) accumulation at 37 days For Se content in the blood, 0.2 ppm in the feed of Selsaf is equivalent to 0.4 ppm of sodium selenite. It means that to obtain the same quantity of Se in the blood, two times less Se-yeast compared to sodium selenite is needed. While to obtain the same level of Se in the pectoral muscle with sodium selenite compared to 0.2 ppm of organic Se in the feed, it would be necessary to add 3.06 ppm of sodium selenite in the feed. But, the addition of such a high level of sodium selenite in the feed is not possible due to the toxicity of Se at high level and the maximum level of Se authorized in feed in the European Union, which is 0.5 ppm of total Se. The difference of Se accumulation in the blood and pectoral muscle between organic and inorganic Se sources can be explained by their mode of absorption. Sodium selenite is absorbed via passive diffusion whereas organic Se like Se-meth is absorbed by active transport (sodium-dependant methionine system). Moreover, the Se-meth will substitute the methionine in the synthesis of all proteins in the pectoral muscle, which explain the better efficiency of organic Se containing mainly Se-meth. Ingested Se-meth is either metabolized directly into reactive forms of Se or stored in place of methionine in body proteins. Se-meth metabolism is closely linked to protein turnover. Although World’s Poultry Science Journal, Volume 69, Supplement 4 EggMeat Symposia 2013 Bergamo 15-19 September 2013 selenite and selenate can be used for selenoprotein biosynthesis, only Se-meth is incorporated into body proteins. As humans or poultry have no efficient mechanism for methionine synthesis, they are also unable to synthetize Se-met. (according to the review of Gerahard N. Schrauzer, 2000) The difference of efficiency among Se-yeast sources for pectoral muscle accumulation can be due to their different composition and mainly their different content in Se-meth: the Selsaf containing 63% of Semeth is more efficient than the other Se-Yeast containing 56.7% of Se-meth. This accumulation of Se in the muscle can be a good source of Se for human nutrition. In fact, depending on the area, some population has some deficiency of Se, which can lead to poor health, genetic defects, decreased fertility and defense against various viral and bacterial diseases (Fisinin et al., 2008). The Recommended Daily Allowance (RDA) in the European Union is 55 µg per day (European directive 90/496/EEC). So, by eating 100 g of meat coming from a broiler supplemented with Selsaf at 0.2 ppm, 47.4 % of RDA are covered. When the meat comes from a broiler supplemented with 0.2 ppm of selenite, only 19 % of RDA is covered. So, Se-enrichment of animal derived foods via supplementation of animal feed can be an effective way of increasing human Se status in countries where Se consumption is below the RDA (Fisinin et al., 2008). Conclusions In conclusion, the use of organic Se in the form of Se-yeast in the feed of broilers allows a better transfer of Se into the blood and in the muscle compared to Sodium selenite. A level of 0.2 ppm of Selsaf is equivalent to 0.4 ppm or 3.06 ppm of sodium selenite for blood Se accumulation and meat Se enrichment respectively. The Se in blood and muscle is well correlated to the quantity of Se in the feed and is dependant on the chemical form of Se. The difference of bioavalaibility between Se sources is due to different mode of absorption in the intestine and different metabolisms. All sources of Se-Yeast are not equal depending on their content in Se-meth. The increase of Se in muscle is of interest for human nutrition and for improving meat quality. References DIRECTIVE 90/496/EEC on Nutrition Labelling for Foodstuffs: Discussion paper on revision of technical issues. (2006) DOWNS KM., HESS JB., BILGILI SF. (2000) Selenium Source Effect on Broiler Carcass Characteristics, Meat Quality and Drip Loss. Journal of Applied Animal Research. 18. FISININ VI., PAPAZYAN TT., SURAI PF. (2008) Producing specialist poultry products to meet human nutrition requirements : Selenium enriched eggs. World’s Poultry Science Association. 64; 8597. KIM YJ., Park WY., Choi IH. (2010) Effects of dietary α-tocopherol, selenium and their different combinations on growth performance and meat quality of broiler chickens. Poultry Science. 89(3); 603-608. ROTRUCK JT., POPE AL., GANTHER HE., SWANSON AB., HAFEMAN DG., HOEKSTRA WG. (1973) Selenium : biochemical role as a component of Glutathione Peroxidase. Science. 179: 588-590. SCHRAUZER NG. (2000). Selenomethionine: A review of its nutritional significance, metabolism and toxicity. Recent advances in nutritional sciences. 1653-1656. SURAI PF. (2002) Selenium in poultry nutrition 1. Antioxidant properties, deficiency and toxicity. World’s Poultry Science Journal. 58; 333-347. WHANGER PD. (2002) Selenocoumpounds in plants and animals and their biological significance. Journal of the American College of Nutrition. 21; 223-232. YOSHIDA M., FUKUNAGA K., TSUCHITA H., YASUMUTO K. (1999) An evaluation of the bioavailability of Selenium in High-Selenium Yeast. J. Nutr. Vitaminol. 45; 119-128. VAMECQ J., VALLEE L., STORME L., GELE P., BORDET R. (2004) Key players in oxidative stress. La lettre du pharmacologue. 18; 16-23. World’s Poultry Science Journal, Volume 69, Supplement 5